Quantitative aspects of red cell size variation during pregnancy

Summary Red cell volume distribution curves may be readily obtained using electronic cell counters, and the proportion of cells that deviate from the normal population may be determined. In this study, we report the results of red cell volume distribution analyses in pregnant and non-pregnant females. The relationship between red cell volume distribution and other red cell indices is also studied. Statistical analysis showed good correlation between various Channelyzer parameters and red cell indices including Hb, RCC, MCV, MCH, and MCHC. A significant proportion of pregnant patients showed abnormal red cell volume distribution curves particularly in the latter half of pregnancy. During the latter half of pregnancy there was an increased proportion of microcytic cells in 8% of patients and evidence for macrocytosis was found in 67% of patients. In general there was good correlation between stages of gestation and Channelyzer parameters associated with macrocytosis. It is suggested that analysis of red cell size, which may be simply performed with minimal quantities of blood and inconvenience using the Channelyzer, could give insight into those conditions associated with variation in red cell size including iron deficiency and folate deficiency in pregnancy and it is conceivable that repeated estimations of these parameters could obviate the need for blanket administration of haematinics to the pregnant patient.     

Simultaneous measurement of reticulocyte and red blood cell indices in healthy subjects and patients with microcytic and macrocytic anemia

Using the new Bayer H*3 hematology analyzer (Leverkusen, Germany), we have determined red blood cell and reticulocyte indices in 64 healthy subjects, in patients with microcytosis due to iron deficiency (58 patients) and heterozygous beta-thalassemia (40 patients), and in patients with macrocytosis (28 patients). We found in all cases that reticulocytes were larger than mature red cells by 24% to 35%, with a hemoglobin concentration 16% to 25% lower and a similar hemoglobin content. The correlation between red cell and reticulocyte indices was strikingly tight (r = .928 for volume, r = .929 for hemoglobin concentration, r = .972 for hemoglobin content) in all four groups, regardless of red blood cell size. The ratio of reticulocyte to red blood cell mean corpuscolar volume (MCV ratio) was constantly above 1. Inversion of the MCV ratio was observed only in four patients. It was always abrupt and transitory and was associated with erythropoietic changes leading to the production of red blood cells of a different volume (treatment of megaloblastic anemia, functional iron deficiency, bone marrow transplantation). In two cases of marrow transplantation, reticulocyte volume fell during the aplastic phase after conditioning chemotherapy and then rapidly increased up to values higher than before; this production of macroreticulocytes was the earliest sign of engraftment.     

Use of advanced red blood cell and reticulocyte indices improves the accuracy in diagnosing iron deficiency in pregnant women at term

Objectives: Detection of iron deficiency during pregnancy with hemoglobin (Hb) and serum measurements is insignificant as the measurements may be affected by e.g. hemodilution or accelerated erythropoiesis. This study tests whether cell indices will give a more reliable measure of iron deficiency in pregnant women at term. Methods: The population was 202 pregnant women. Using the ADVIA 120 hematology system, Hb, mean cell volume (MCV), percentage of hypochromic red blood cells (%HYPOm) and reticulocytes (%HYPOr), and cellular hemoglobin in reticulocytes (CHr) were tested. Additionally, transferrin saturation (TfSat), ferritin, and transferrin receptor (TfR) were analyzed. Receiver operating characteristic (ROC) curves and area under the ROC curves (AUC) were used as statistical methods. Results: When TfSat (≤11%) was used as the reference test for iron deficiency, %HYPOm and CHr had a sensitivity of 58.1% and 80.7%, while the specificities were 82.6% and 71.3%, respectively. Additionally, the AUC values were %HYPOr 0.80, CHr 0.79, ferritin 0.77, %HYPOm 0.75, TfR 0.67, MCV 0.63 and Hb 0.64. The results provided by the cell indices alone (%HYPOm or CHr) were in good agreement with the results based on the usage of a combination of three commonly used tests (Hb, MCV, ferritin). Conclusions: This study suggests that the most practical way to diagnose iron deficiency in pregnant women at term is to use cell indices such as CHr and %HYPOm provided by the automated hematological analyzer. Further studies are needed to determine the usefulness of the cell indices in diagnosing iron deficiency longitudinally during the course of pregnancy.     

Evaluation of Macrocytosis in Routine Hemograms

Macrocytosis, a condition in which erythrocytes are larger than normal manifests as an increase in mean corpuscular volume (MCV) more than 100 fl. The aim of this study was to identify the underlying causes of macrocytosis, detected in routine hemograms and to evaluate the hematological features in different etiologies. This study included 178 adult patients whose detailed medical history was recorded, and Vitamin B12 assay, folate assay, thyroid function tests, liver function tests, complete blood counts and peripheral smear evaluation was performed. Alcoholism was identified as the etiological factor in 65 cases (36.5%), Vitamin B12 deficiency in 43 cases (24.1%) and drug related in 23 cases (12.9%). These three conditions accounted for 73.6% of macrocytosis. Other causes identified were folate deficiency, liver disease, Myelodysplastic syndrome, chronic renal failure and Aplastic anemia. In 41 cases, the cause of macrocytosis could not be explained. Anemia was observed in 95 cases (53.3%) being most common in Vitamin B12 deficiency. 9 cases (20.9%) of Vitamin B12 deficiency presented with isolated macrocytosis without anemia. It was observed that mean hemoglobin was lower and red cell distribution width (RDW) higher in megaloblastic conditions. Peripheral smear revealed hypersegmented neutrophils in 86% and macro-ovalocytes in 72% of the megaloblastic cases. Complete medical history, red cell parameters and peripheral blood smear are simple, inexpensive tools which assist in identifying the underlying cause of macrocytosis, particularly in resource limited settings. Macrocytosis needs to be evaluated even in the absence of anemia, as it may be the first clue to an underlying pathology.     

Red cell macrocytosis in hypoxemic patients with chronic obstructive pulmonary disease

Macrocytosis is a common finding in patients with chronic obstructive pulmonary disease (COPD). The cause for the elevation of mean corpuscular volume (MCV) in these patients remains elusive. In an attempt to determine the extent of macrocytosis in COPD patients and search for possible causative factors, we evaluated the hematologic parameters, F-cell percentage, blood gases and serum erythropoietin (Epo) Levels in 32 COPD clinically stable patients and 34 sex- and age-matched non-smoker healthy volunteers. An increased MCV was observed in almost half of the hypoxemic COPD cases (14/32 or 43.75%), while erythrocytosis developed to a lesser degree (37.5%). The erythropoietic response did not correlate with the severity of hypoxia. Moreover, no significant correlation was found between macrocytosis and hypoxemia or erythrocytosis and red cell size. In some cases the two phenomena occurred independently. The F-cell percentage was significantly elevated in the COPD group (P < 0.01) and was associated with MCV values (n = 32, r5 = 0.41, P < 0.05). This finding supports the hypothesis we put forward to explain the macrocytosis often observed in COPD, i.e., that the acute erythropoietic stress occurring repeatedly in these patients as a result of the frequent exacerbations may lead to waves of release of relatively immature, large red cells from the marrow, including an increased number of F-cells, reflecting the recruitment of normally dormant BFU-E (bursts forming units of erythrocyte precursors), which maintain the program for gamma-chain synthesis. The fact that erythrocytosis and macrocytosis, both being triggered by hypoxemia, do not occur consistently in all COPD patients indicates that many other factors may also intervene.     

Absence of macrocytic anaemia in Alzheimer's disease

There is an association between Alzheimer's disease (AD) and low serum levels of vitamin B12 and folic acid. Patients also have elevated serum levels of homocysteine and disease progression might therefore be associated with the development of a macrocytic anaemia. We investigated the relationship between disease duration, homocysteine and haematological indices in patients with clinically diagnosed AD and healthy elderly controls. Haemoglobin and platelet counts fell only slightly with increasing dementia duration, but there were no other changes in haematological indices. In particular, macrocytosis and red cell distribution width were unrelated to disease duration and no patients were anaemic. Our results support previous observations that the neurological and haematological features of B12 and folate deficiency are often unrelated in these patients.     

Erythropoiesis during recovery from macrocytic anemia: macrocytes, normocytes, and microcytes

In seven patients with marked megaloblastic anemia (MCV greater than 110 fl), red cell size distribution curves (erythrograms) demonstrated the size of red cells produced after therapy. In six, the new red cells were normocytic throughout recovery. In the seventh patient, folate repletion along produced a new population of microcytes, due to unsuspected iron deficiency; after iron repletion normocytes were produced. Three patients with autoimmune hemolytic anemia had macrocytosis (MCV greater than 110 fl) without folate or vitamin B12 deficiency. During recovery with predisone therapy, instead of a discrete new normocytic population appearing, the entire population progressively returned to normal size. Normal rather than "stress" reticulocytes, and remodeled stress reticulocytes remaining, may explain this different pattern of recovery. Two patients initially had minor subpopulations of smaller red cells that disappeared soon after therapy. These probably reflected the dyserythropoiesis of severe megaloblastic anemia.     

Spurious automated red cell values in warm autoimmune hemolytic anemia

Two patients with warm autoantibodies to their red cells had a mean cell volume artifactually elevated by red cell agglutination. Red cell size distribution histograms directly showed doublets and triplets of normal-size red cells. This phenomenon is similar to the spurious macrocytosis previously reported due to cold agglutinins, but was not reversible by warming.     

Discriminant analysis of macrocytic red cells

Summary. Laboratory classification of red cell disorders uses the red cell indices (MCV, MCH, MCHC, RDW) and information gleaned from microscopic evaluation of a blood film. Additional red cell information is now available using the H series of automated blood cell analysers (Ames Technicon Division of Bayer Diagnostics). This study involved the development of a discriminant rule which would differentiate between three causes of macrocytosis (vitamin B₁₂/folate deficiency, alcohol excess/liver disease and a reticulocytosis) using the information available on Research Screen 1 and Report Screen 3 of the H*1 instrument (Report Screen 3 is a graphical display of the trimmed scattergram of red cell volume and red cell haemoglobin concentration and Research Screen 1 displays the associated numerical data). Three methods of analysis were assessed to define a suitable discriminant rule. The percentages of patients correctly classified by the three methods were: 92.1%, 82.0% and 89.2% for Methods 1, 2 and 3 respectively. Method 1 involved the application of quadratic discrimination to transformed variables and produced the best results. Although complex, it could easily be applied using the microprocessor capability of the average multiparameter haematology analyser.     

Diagnostic clues to megaloblastic anaemia without macrocytosis

Masking of the macrocytic expression of megaloblastic anaemia (MA) by coexisting thalassaemia, iron deficiency and chronic illness has been widely reported. We described the haematological and clinical features of 20 Chinese patients with MA presenting with mean corpuscular volume (MCV) < or =99 fl, and analysed the steps leading to the final diagnosis of MA with concomitant thalassaemia trait (n = 11), thalassaemia trait and iron deficiency (n = 3), iron deficiency (n = 4) and chronic illness (n = 2). We also compared the haematological characteristics of this group of patients with a group of normocytic anaemic patients without vitamin B(12)/folate deficiency, and identified certain laboratory information useful for differentiating the two groups. Statistically significant parameters included the mean values of haemoglobin, MCV, red cell distribution width (RDW), reticulocyte index, platelet count and serum bilirubin. All provided clues to maturation disorders within the marrow. A decision flowchart for the diagnosis of MA without macrocytosis was proposed. In the studied population, by using the parameters of haemoglobin <10 g/dl, MCV 80-99 fl, RDW > or = 16% and reticulocyte index < or = 2% as indicators, there was a 58% chance that a patient had MA without macrocytosis if he/she had all the four indicators, and a 2.2% chance of having it if he/she did not have these indicators. We emphasized the importance of including peripheral blood smear examination in the diagnostic procedures for such patients, as well as the importance of paying attention to patients' medical history, racial background and previous MCV value.     

The relationship between serum cobalamin concentration and mean red cell volume at varying concentrations of serum folate

There is concern that exposure of patients to folic acid may prevent the development of the macrocytosis of cobalamin deficiency and thus delay the detection of the neurological complications. We examined the relationship between low cobalamin levels and mean cell volume (MCV) at different serum folate concentrations in 63,472 blood samples tested in a community pathology laboratory over 2 years. We found no evidence that high serum folate levels masked the macrocytosis of cobalamin deficiency in this population with similar increases in MCV regardless of whether the serum folate was low, normal or high. Macrocytosis appears to retain its value as a marker of cobalamin deficiency in people with serum folate concentrations above the population average.     

Clinical implication of hematological indices in the essential hypertension

Prognostic value of haematological indices,especially red cell distribution width,neutrophil lymphocyte ratio and mean platelet volume,was reported with numerous investigations in miscellaneous cardiovascular settings.Their major prognostic value was linked to oxidative stress and inflammation since their level was correlated with major inflammatory markers such as high sensitive C-reactive protein and interleukins.Oxidative stress and chronic inflammation are also postulated as the main pathophysiologic mechanism of essential hypertension(HT) and its vascular complication.Recently,correlation between HT and haematological parameters was searched in numerous studies,which has made the topic more popular.Herein,we reveal the correlation between haematological indices and HT and we also demonstrate the clinical implication of this correlation.Impaired haematological parameters may strongly indicate hypertensive end-organ damage.     

Erythropoiesis during recovery from iron deficiency: normocytes and macrocytes

In 26 patients with severe iron deficiency and microcytic anemia (MCV less than 70 fl), serial red cell size distribution histograms (erythrograms) were taken before and during iron therapy. Initially all patients had a single population of red cells, all microcytes. With the first reticulocytosis after iron therapy, a new population of cells appeared, larger in volume than the original. In 23 of 26 patients the new population of cells was of normal size (82-96 fl). In 3 of 26, the new population was macrocytic (MCV greater than 98 fl). Of these 3, 1 had folate deficiency; after folate was given, normocytes were produced. The other 2, both taking phenytoin and 1 a heavy alcohol using, had persistent macrocytosis despite folate administration. Erythrograms allowed quantitative, rapid evaluation of erythropoietic response to iron repletion. Abnormal macrocytic responses could be identified and seemed to occur with some frequency.     

Platelet indices in chronic alcoholic liver disease patients with thrombocytopenia

AIM: To detect alterations in platelet indices in patients with chronic alcoholic liver disease and thrombocytopenia, and its correlation with other haematological parameters. METHODS: We studied 65 individuals separated in two groups: controls (n = 35) and chronic alcoholic liver disease patients with thrombocytopenia (n = 30). The control group was age and gender matched with patients group. In all, controls and patients, a haematological evaluation was done, including platelets indices. RESULTS: In the patients group we found a low number of erythrocytes, leucocytes and platelet when we compare with controls. The same is true when we compare haemoglobin, hematocrit and absolute count of lymphocyte and neutrophil. The mean globular volume, mean globular haemoglobin and red cell distribution width where significantly higher in patients group. Platelet indices showed a statistical significant increased in platelet distribution width and decreased in platelet crit in the patient group. No differences where found on mean platelet volume between the two groups. Correlation between platelet number and other haematological parameters was found. CONCLUSION: Chronic alcoholic liver disease patients showed a decrease in all haematopoietic cell lines, probably associated with hypersplenism found in those patients. Additionally to the numeric alterations the erythrocyte and platelets showed morphologic alteration revelled by respective indices.     

Macrocytosis associated with monoclonal gammopathy

OBJECTIVE: To report the potential association between unexplained macrocytosis and monoclonal gammopathy. METHODS: We retrospectively reviewed the medical records of patients who had monoclonal protein detected by serum electrophoresis and immunofixation from October 1999 until September 2003 at our institution. Patients with concomitant macrocytosis were included in this study. We collected data on patient demographics, evaluations performed for macrocytosis, pertinent laboratory tests relevant to the diagnosis of monoclonal gammopathy and presence of associated hematologic disorders. RESULTS: We identified 258 patients with monoclonal gammopathy. Thirty-one (12%) of them had concomitant macrocytosis. Of the latter group, 14 (5%) patients had no identifiable cause of macrocytosis after thorough evaluation and were considered to have macrocytosis associated with monoclonal gammopathy. The median values for mean cell volume and serum monoclonal protein were 103.9 fL (range 100.8-109.8) and 1.95 gm/dL (range 0.8-4.3), respectively. Most patients had IgG (71%) and kappa light chain (79%). All of the 11 (of 14) patients who underwent a bone marrow biopsy as part of the initial evaluation had megaloblastoid maturation of the erythroid precursors. No correlation was found between the level of serum monoclonal protein and the degree of macrocytosis (r = +0.48, P = 0.08). After a median follow-up of 22.5 month (range 3-60+), all but one patient had persistent but stable macrocytosis. CONCLUSION: Macrocytosis can be a manifestation of monoclonal gammopathy. Disorders associated with monoclonal gammopathy should be considered in the differential diagnoses during evaluation of macrocytosis.     

Microcytosis, Anisocytosis and the Red Cell Indices in Iron Deficiency

S ummary . Red cell volume distribution curves have been used to measure micro-cytosis and anisocytosis in normal subjects, blood donors and patients with iron deficiency anaemia. These measurements were more sensitive than the conventional red cell indices for detecting blood donors with a low transferrin saturation. Three stages are suggested as iron deficiency progressively interferes with haemopoietic function. Anisocytosis and an increased percentage of microcytic cells are the first haematological abnormalities to occur and at this stage haemoglobin concentration is usually normal and transferrin saturation less than 32%. At the second stage the MCV and MCH decline, haemoglobin concentration is generally sub-normal, though not below 9 g/dl, and transferrin saturation is usually below 16%. The final stage of iron deficiency is associated with a low MCHC, a haemoglobin concentration below 9 g/dl and a transferrin saturation of less than 16%.     

Folate Deficiency in the Alcoholic—its Relationship to Clinical and Haematological Abnormalities, Liver Disease and Folate Stores

Clinical and laboratory observations were made on 84 patients regularly taking more than 80 g of alcohol daily. Macrocytosis was present in 84.5%, but only 13% were anaemic. Twenty-seven of the 57 bone marrows sampled were abnormal, 20 showing megaloblastic changes, mostly mild in degree. Serum, red cell, and liver folate levels were reduced in 28%, 35% and 31% of patients respectively. Liver folate concentration showed good correlation with serum and red cell folates. The folate deficiency was more severe in those patients, more often female, who had inadequate diets or drank wine and spirits rather than beer. The amount of alcohol consumption and extent of liver damage did not affect folate status. The findings suggest that folate deficiency is common among alcoholics in this country. More frequently, however, patients had macrocytosis, sometimes with megaloblastosis, in the absence of folate deficiency emphasizing the direct toxic effect of alcohol on the developing erythroblast.     

Whole blood viscosity and erythrocyte hematometric indices in chronic heroin addicts

Whole blood viscosity (WBV) and hematometric indices of erythrocytes as red blood cell count (RBC), mean erythrocyte volume (MCV), hemoglobin (HGB), hematocrit (HCT), mean hemoglobin content of erythrocytes (MCH), HGB/HCT values (MCHC) and red blood cell distribution width (RDW) have been studied in a group of 15 chronic opioid addicts under methadone maintenance therapy with mean age 26.53 +/- 7.34 years. WBV elevation and changes in MCV, HGB, HCT, RDW were found in intravenous drug users compared to healthy individuals. As well, RBC was decreased leading to an increase in MCH and MCHC values. Correlation analysis suggested that the correlation among the RBC, HGB, HCT and WBV was the closest. Heroin macrocytosis (heroin macrocytic anemia) was established, related with the increased RDW in chronic heroin abusers. The results are in accordance with data revealing abnormal effects of alcohol and other drugs on whole blood rheology and hematometric/morphometric characteristics of erythrocytes.     

Relationship of lymphocyte anisocytosis in chronic lymphatic leukaemia to mouse red cell rosetting capacity and clinical stage

In 34 patients with chronic lymphatic leukaemia (CLL) the lymphocytes have been separated and sized using a C1000 Channelyzer. The modal volume and the volume range of the populations have been obtained and related to clinical stage and mouse red blood cell (MRBC) rosetting capacity. Over 1 year's observation with several estimations per patient there was no convincing drift towards increase in modal volume with deteriorating clinical status. The cell size of the CLL populations could vary from time to time in the same patient. The MRBC rosetting capacity varied greatly between estimations. The findings suggest that in CLL there is an oscillation in cell size in a given patient when tested at intervals.     

Irreversibly sickled cells and red cell survival in sickle cell anemia: a study with both DF32P and 51CR

For 25 subjects with sickle cell anemia the mean red cell life span measured with Di-isopropylfluorophosphite-32P (DF32P) was 17.32 +/- 4.51 days. Performed simultaneously, the half life (T1/2 of radioactively-labelled chromium 51Cr) was 10.11 +/- 2.82 days (14 subjects). Eight additional subjects, or more than 30 per cent of those studied using both red cell tags, had 51Cr red cell survival curves better described by two exponents than by one, apparently due to two different rates of 51Cr elution from the red cells. This finding limits the value of quantitative data obtained by this procedure. A negative correlation was found between the mean red cell life span measured with DF32P-tagged cells and the proportion of irreversibly sickled cells in venous or capillary blood. A similar negative correlation was found between the red cell half survival time measured with 51Cr-tagged cells and the proportion of irreversibly sickled cells. These data are compatible with the view that repeated sickling and, in particular, the formation of irreversibly sickled cells play a distinct role in the pathogenesis of hemolysis in sickle cell anemia.