Human leucocyte antigens A,B, C,and DRW in idiopathic "warm" autoimmune haemolytic anaemia.

Twenty patients with idiopathic "warm" autoimmune haemolytic anaemia and 40 controls were types concurrently for human leucocyte antigens (HLA) A, B, C, and DRW. There was a significantly stronger association of HLA-B8 with the disease (chi 2 = 10.39; p = 0.018) than HLA-DRW3 (chi 2 = 3.71; P = 0.35) and the patients also showed a significant increase in BW6 homozygosity (chi 2 = 7.13; P = 0.01) and a corresponding reduction in BW4 (chi 2 = 7.13; P = 0.02). (All p values corrected for number of antigens at each locus.) These findings suggest that susceptibility to idiopathic autoimmune haemolytic anaemia is associated more closely with the HLA-B locus than with DRW3.     

Drug-induced Blood Dyscrasias in Sweden

Cases of drug-induced aplastic anaemia, haemolytic anaemia, thrombocytopenia, and agranulocytosis reported to the Swedish Adverse Drug Reaction Committee during the five-year period 1966-70 have been analysed and compared with cases of the same cytopenias from “all” causes. Oral diuretics were a dominant cause of drug-induced thrombocytopenia, methyldopa of haemolytic anaemia, and oxyphenbutazone of aplastic anaemia. Computer systems should help such studies, particularly in showing a changing pattern of complications and causes.     

Screening for hemochromatosis in routine medical care: an evaluation of mean corpuscular volume and mean corpuscular hemoglobin

Our aim was to evaluate the potential utility of mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) to detect hemochromatosis. We computed the accuracy of MCV and MCH cut-off points > or = upper reference limits using data from 94 probands and 132 white controls. Our reference ranges are MCV 80.0-97.0 fL and MCH 26.0-32.0 pg. Sensitivity of MCV was 8.6-48.3% for men and 2.8-44.4% for women (cut-off points > or = 105.0 - > or = 97.0 fL, respectively). Sensitivity of MCH was 33.9-70.7% for men and 19.6-50.0% for women (cut-off points > or = 34.0 - > or = 32.0 pg, respectively). Using MCV and a hemochromatosis frequency typical of many western Caucasian populations (0.005), positive predictive values (PV+) were 2.1-100.0% in men and 4.2-100.0% in women. Using MCH, PV+ were 1.7-8.2% in men and 1.8-6.8% in women. We also calculated PV+ using the hemochromatosis frequency 0.015, which could occur in persons receiving medical care. Using MCV cut-off points > or = 101.0 fL, PV+ were 8.9-100.0% in men and 100.0% in women with maximum sensitivities of 24.1% and 25.0%, respectively. Using MCH testing, PV+ was 21.5% in men (cut-off point > or = 34.0 pg) and 18.2% in women (cut-off point > or = 33.0 pg) with sensitivities of 33.9% and 37.0%, respectively. Using MCV or MCH, sensitivity and PV+ for the HFE genotype C282Y/C282Y were generally greater than for "nonclassical" HFE genotypes. All negative predictive values in our study were > or = 98.5%. We conclude that supranormal values of MCV or MCH could be used to detect hemochromatosis in white persons of western European descent who are receiving routine medical care. Comparisons of MCV, MCH, and transferrin saturation testing and other implications of MCV and MCH testing for hemochromatosis in medical care are discussed.     

Behaviour of cell physiological and biochemical indicators of maturation and aging of red blood cells in density fractionated red blood cells during a bleeding anaemia of the rabbit

Changes of the density, MCV, MCHC, RNA-concentration and the activities of the enzyme G6PD and ASAT due to maturation and aging of rbc were investigated by repeated density gradient separation of these cells during a bleeding anaemia of the rabbit. The mean density of reticulocytes and erythrocytes decreases during the course of the anaemia by 14 and 10 g/l respectively. The difference between the density of both cell types increases from 3 to 6 to 12 g/l during the anaemia. Maturation and aging of rbc are characterized by an increase of the MCHC from 200 to 350 g/l cells and the decrease of MCV by 10-25%. The RNA concentration and the ASAT activity show an almost parallel behaviour in cells of different densities during the anaemia. Remarkable differences are observed between the ASAT activity and the G6PD activity: The G6PD activity decreases mainly during the aging process and almost not during the maturation, while the ASAT activity falls constantly during maturation and aging. As observed in former studies we could distinguish and isolate 4 characteristic populations of reticulocytes in certain density fractions during the anaemia: Normal young reticulocytes, line 2 reticulocytes, megaloreticulocytes and reticulocytes of high density.     

Isoimmune haemolysis in pathogenesis of anaemia after cardiac surgery.

A patient who had received multiple transfusions developed antiglobulin-positive haemolytic anaemia due to a delayed haemolytic transfusion reaction. Many cases of haemolytic anaemia after cardiac surgery could be explained on this basis.     

Microangiopathic hemolytic anemia in patients with diseases of the hematopoietic and lymphatic systems

Microangiopathic haemolytic anaemia was diagnosed in the course of haematopoietic and lymphatic disorders such as chronic granulocytic leukemia, chronic myelofibrosis, chronic lymphatic leukemia, Osler's disease, chronic monocytic leukemia, and lymphoplasmocytic lymphoma, in 11 patients (6 women and 5 men) aged between 33 and 81 years (mean age 58.8 years) treated at the Haematological Out-Patient Clinic of the Postgraduate Medical Education Centre within 1977-1987. The following laboratory tests were carried out: 1) morphology of the peripheral blood and bone marrow, especially some haematological parameters concerning erythrocytes and blood platelets; 2) biochemical tests reflecting erythrocytes disintegration; 3) haemostasis. All examined patients suffered from haemolytic anaemia of various degree with characteristic changes in erythrocyte shape (helmets, tear-drops etc.). Haemolytic origin of anaemia was confirmed by the increased LDH activity. In the majority of patients no compensative stimulation of haematopoiesis (reticulocytosis, red blood cells hyperproliferation in bone marrow) was seen. Clinical symptoms of haemostatic disorders such as haemorrhagic diathesis and vein thrombosis were diagnosed in 50% of the patients. Blood platelet counts ranged from markedly decreased to significantly increased. Bone marrow smears did not show increased number of megacariocytes. Bleeding time was prolonged in the majority of examined patients while prothrombin index--decreased). Abnormal fibrinogen levels (decreased or increased) were found in the majority of patients with fibrin degradation products. Microangiopathic haemolytic anaemia in these patients differ from the typical Moschowitz's disease clinically probably due to the lack of compensative stimulation of erythropoiesis and lower thrombocytopenia.     

THE NUMBER AND POSSIBLE FUNCTIONS OF DNA-SYNTHESIZING CELLS IN HUMAN BLOOD

The number of DNA-synthesizing cells in the blood of patients with various disorders was studied autoradiographically after incubation of blood in vitro with [³H]thymidine. The DNA-synthesizing cells were cytologically assigned to the following categories: erythroid, myeloid, lymphoplasmacytoid and unidentifiable (monocytoid or blast-like) cells. The following patient categories were studied: mitral valvular disease (samples obtained from peripheral vein, pulmonary artery and left auricle), ‘autoimmune diseases’(systemic lupus erythematosus, schleroderma, Hashimoto's thyroiditis, immunohaemolytic anaemia), patients with depressed haemopoiesis (aplastic anaemia, nitrogen-mustard induced bone-marrow hypoplasia) and with increased haemopoiesis (haemolytic anaemia, pernicious anaemia before and during initial vitamin-B₁₂ therapy, red-cell mass regeneration after haemorrhage or iron deficiency) and patients with bacterial infection. In all conditions studied, the number of labelled monocytoid and blast-like cells varied between 0 and 4/μl. Similarly, the number of labelled lympho-plasmo-cytoid cells was consistently low (0–8/μl) in all cases studied except two, where values of 37 and 63/μl were found. Both these patients had severe bacterial infections. The function(s) and potential(s) of these cells are discussed. The fate of the blast-like and monocytoid cells remains obscure. The lympho-plasmocytoid cells probably serve an immunological function, perhaps by disseminating immune responses. Whether or not some DNA-synthesizing cells in the blood are haemopoietic stem cells cannot be decided from the available evidence.     

Haemolytic anaemia after cisplatin treatment.

Normochromic or normocytic anaemia is a common side effect of treatment with cisplatin. Two patients treated with cisplatin 100 mg/m2 in combination with vinblastine, bleomycin, and actinomycin D developed haemolytic anaemia. Neither patient had evidence of haemolysis before treatment, and in both cases severe haemolytic anaemia developed after several courses of cisplatin and when the cancer had regressed almost completely. The importance of haemolysis in the development of anaemia after cisplatin treatment has not been investigated fully and further studies are needed.     

Experimentally induced toxic-haemolytic anaemia in laboratory rats following phenacetin administration

In an attempt to give assistance in the search for a simple haematological system for the detection of toxic-haemolytic effects of chemical substances, we used a battery of simple tests and phenacetin as a reference substance. The drug was administered in a single dose or repeatedly for two or four weeks. Higher plasma haemoglobin levels and symptoms of anaemia (decreased blood haemoglobin levels, erythrocytic counts, haematocrit) and activated erythropoiesis (increased reticulocyte counts, MCV, marrow and splenic E:M ratios) were found to be best marked following administration of phenacetin in the very high dose of 1.0 g/kg per orally for two weeks. Our results suggest that plasma haemoglobin levels could provide information on the haemolytic potential of phenacetin if the experimental schedule included the optimal combination of suitable phenacetin dosage and duration of loading.     

Increased erythrocyte adenosine deaminase activity without haemolytic anaemia

A 4-fold increase of red blood cell adenosine deaminase (ADA) activity was found in a patient without haemolytic anaemia, but with mild anisopoikilocytosis. High-performance liquid chromatography showed a 40% reduction of adenosine-5'-triphosphate (ATP) while all the other nucleotides were in normal ranges. The patient's parents (first cousins) and a brother displayed the same enzyme activities as the controls. This observation suggests that mild increases of ADA activity is neither a marker for congenital hypoplastic anaemia as previously reported nor associated with haemolytic anaemia.     

Severe anaemia is associated with a higher risk for preeclampsia and poor perinatal outcomes in Kassala hospital,eastern Sudan

Background

Anaemia during pregnancy is major health problem. There is conflicting literature regarding the association between anaemia and its severity and maternal and perinatal outcomes.

Methods

This is a retrospective case-control study conducted at Kassala hospital, eastern Sudan. Medical files of pregnant women with severe anaemia (haemoglobin (Hb) < 7 g/dl, n = 303) who delivered from January 2008 to December 2010 were reviewed. Socio-demographic and obstetric data were analysed and compared with a similar number of women with mild/moderate anaemia (Hb = 7-10.9 g/dl, n = 303) and with no anaemia (Hb > 11 g/dl, n = 303). Logistic regression analysis was performed separately for each of the outcome measures: preeclampsia, eclampsia, preterm birth, low birth weight (LBW) and stillbirth.

Results

There were 9578 deliveries at Kassala hospital, 4012 (41.8%) women had anaemia and 303 (3.2%) had severe anaemia. The corrected risk for preeclampsia increased only in severe anaemia (OR = 3.6, 95% CI: 1.4-9.1, P = 0.007). Compared with women with no anaemia, the risk of LBW was 2.5 times higher in women with mild/moderate anaemia (95% CI: 1.1-5.7), and 8.0 times higher in women with severe anaemia (95% CI: 3.8-16.0). The risk of preterm delivery increased significantly with the severity of anaemia (OR = 3.2 for women with mild/moderate anaemia and OR = 6.6 for women with severe anaemia, compared with women with no anaemia). The corrected risk for stillbirth increased only in severe anaemia (OR = 4.3, 95% CI: 1.9-9.1, P < 0.001).

Conclusions

The greater the severity of the anaemia during pregnancy, the greater the risk of preeclampsia, preterm delivery, LBW and stillbirth. Preventive measures should be undertaken to decrease the prevalence of anaemia in pregnancy.

     

Assessment of iron and zinc intake and related biochemical parameters in an adult Mediterranean population from southern Spain: influence of lifestyle factors

This study assessed factors influencing iron and zinc intake and associated biochemical parameters in an adult population from southern Spain to identify patterns of intake and groups at risk for deficiency. A cross-sectional survey was done in Andalusia (southern Spain) to study nutrient intakes in a random sample of 3421 subjects (1747 men, 1674 women, age between 25 and 60 years). Blood samples were obtained for biochemical assays in a random subsample of 354 subjects (170 men, 184 women). Food consumption was assessed by 48-h recall. In blood samples, we measured red blood cells, haemoglobin (Hb), haematocrit, total iron binding capacity (TIBC) and plasma concentration of Fe and Zn. Information about educational level, smoking habit, alcohol consumption and physical exercise was collected with a structured questionnaire. Intakes were below two thirds of the recommended dietary allowances (RDA) in 22.45% of the sample for Fe and in 56.45% for Zn. Iron deficiency [two or more abnormal values for plasma Fe, TIBC, transferrin saturation and mean cell volume (MCV)] was found in 12.7% of the sample, and iron-deficient anaemia (low values for Hb, MCV, mean cell Hb and mean corpuscular Hb concentration) was found in 2.1%. In smokers, plasma levels of Fe were higher, and MCV was lower than in nonsmokers. Plasma Zn concentrations were below the reference value in 17.8% of the persons. Age and body mass index correlated inversely with plasma Zn (P<.01). Gender, age, obesity, smoking, alcohol consumption and physical activity were associated with differences in nutrient intakes. Logistic regression analysis showed that female gender and older age were associated with the risk of low intakes of Fe and Zn.     

Sustainable malaria control: transdisciplinary approaches for translational applications

Background

Intravenous (IV) artesunate is the treatment of choice for severe malaria. In Europe, however, no GMP-manufactured product is available and treatment data in European travellers are scarce. Fortunately, artesunate became available in the Netherlands and Belgium through a named patient programme. This is the largest case series of artesunate treated patients with severe malaria in Europe.

Methods

Hospitalized patients treated with IV artesunate between November 2007 and December 2010 in the Netherlands and Belgium were retrospectively evaluated. Patient characteristics, treatment and clinical outcome were recorded on a standardized form and mortality, parasite clearance times and the occurrence of adverse events were evaluated.

Results

Of the 68 treated patients, including 55 with severe malaria, two patients died (2/55 = 3.6%). The mean time to 50% parasite clearance (PCT50), 90% and 99% were 4.4 hours (3.9 - 5.2), 14.8 hours (13.0 - 17.2), and 29.5 hours (25.9 - 34.4) respectively. Artesunate was well tolerated. However, an unusual form of haemolytic anaemia was observed in seven patients. The relationship with artesunate remains uncertain.

Conclusions

Data from the named patient programme demonstrate that IV artesunate is effective and well-tolerated in European travellers lacking immunity. However, increased attention needs to be paid to the possible development of haemolytic anaemia 2-3 weeks after start of treatment. Treatment of IV artesunate should be limited to the period that IV treatment is required and should be followed by a full oral course of an appropriate anti-malarial drug.     

Experimental studies on anaemia in sheep infected with Dictyocaulus filaria

The pathogenesis of Dictyocaulus filaria induced anaemia was experimentally investigated. Nineteen Dorset-Muzaffarnagri male lambs were divided into two groups of 13 and six. The lambs in the former group received a primary infection dose of 2000 D. filaria infective larvae, whereas animals in the latter group were kept as uninfected controls. The haematological alterations and faecal larval output were monitored weekly until day 220 post-infection. The infection caused an increase in erythrocyte sedimentation rate (ESR) and total leukocyte count (TLC). However, a decrease in packed cell volume (PCV), haemoglobin, total erythrocyte count (TEC) and blood pH was observed in acute infection. The shape and size of the erythrocytes, serum bilirubin level, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) remained unaffected In carriers, except for TEC, the values of these parameters returned to near-normal levels. The analysis of the results suggests that the infected animals developed a normocytic normochromic anaemia which persisted during the later stage of infection.     

The mean red cell volume in long distance runners

Red cell indices were determined in 6 well trained runners before and after a 100 km race, and Coulter Counter (CC) determinations compared with calculated values derived from centrifuged hematocrit (ctrf), red cell count (CC) and hemoglobin measurements. The following changes were observed immediately after the race, as compared to values 3 days before: MCV(ctrf) decreased by 4.9% (p less than 0.001), MCV(CC) increased by 1.9% (p less than 0.05), MCHC(ctrf) increased by 4% and MCHC(CC) decreased by 3%. The increase in MCV(CC) suggests that intraerythrocyte osmolality was increased, this probably leading to swelling of the cells induced by a shift of water from the diluting Coulter Counter solution into the red cells prior to the MCV measurement. The decrease in MCV(ctrf) immediately after the race was not correlated with the increase in plasma osmolality. This suggests that plasma osmolality alone was not the key factor for regulation of red cell volume. The changes in MCV(ctrf), which contributed to a surprising stability of the hematocrit value and plasma volume, might represent a physiological principle for the maintenance of a favourable blood viscosity.     

Penicillin induced Haemolytic Anaemia

The case histories of two patients with penicillin-induced haemolytic anaemia are presented. One had received 20 mega units a day for 18 days, the other had received 20 mega units a day for two days and then 12 mega units a day for 25 days, before the haemolytic anaemia was diagnosed. Both had previously had courses of penicillin. A strongly positive direct antiglobulin reaction which appeared to be mainly due to IgG antibody was one of the main diagnostic features, and free IgG antipenicillin antibody was found in the serum of both patients. The haemolysis appeared to Lessen as soon as the drug was stopped, and the direct antiglobulin test became negative in 66–77 days.Twelve additional reported cases are reviewed. All had received high doses of penicillin and all had had penicillin previously. The lowest dose recorded was 10 mega units a day for 26 days. The incidence of anti-penicillin antibodies in a hospital population is given, and the mechanism of this type of haemolytic anaemia is discussed. Penicillin-induced haemolytic anaemia should be suspected in any patient receiving penicillin in high doses in whom there is a fall in the haemoglobin level.     

Differences in rat rbc cytosol induced after in vivo phenylhydrazine treatment

Analysis of rat red blood cell (RBC) cytosol protein fraction after in vivo phenylhydrazine treatment revealed increased amounts of 68-kDa protein. This protein is present in trace amounts in normal rat RBC cytosol. We also present data that 68-kDa protein from RBC cytosol has an identical isoelectric point, partial proteolytic map and immunological determinants as a protein of the same molecular mass from rat exosomes. These data indicate that 68-kDa protein is normally present in rat RBC cytosol, is exported via exosomes during reticulocyte maturation, and is increased in induced haemolytic anaemia.     

Age-related sensitivity of rats to induction of anaemia

The intensity of symptoms of experimental anaemia significantly depended on the age of animals. It increased in elder rats after administration of substances with haemolytic potential (phenylhydrazine, phenacetin, 3-chloro-4-benzyloxyphenylacetic acid, nitrobenzene) whereas it decreased after cyclophosphamide-induced erythropoietic suppression.     

Apparent diffusion limitations on branchial CO2 transfer are revealed by severe experimental anaemia in brown bullhead (Ameiurus nebulosus)

In this study, the rapid (within 2 h) effects of acute anaemia on blood gas and acid-base status, as well as cardiorespiratory variables, were examined in brown bullhead (Ameiurus nebulosus). Anaemia was induced by blood withdrawal coupled to volume replacement with saline. Lowering haematocrit from the control value of 23.5+/-1.0% (mean+/-S.E.M.; N=37) to 5.9+/-0.3% (N=37) resulted in a significant increase (by 2.63+/-0.51 torr; N=7) in arterial CO(2) tension (PaCO(2)) over the subsequent 2-h period in the absence of a change in arterial O(2) tension (PaO(2)). Treatment with bovine carbonic anhydrase (CA) reduced the extent of the PaCO(2) increase to the point where it was not statistically significant. In both control and CA-treated fish, arterial pH decreased during acute anaemia; the acidosis was of mixed respiratory and metabolic origin in control fish and primarily metabolic in CA-treated fish. Inducing anaemia caused increases in both cardiac output (V*b) and heart rate that were similar in control and CA-treated fish. Experimental elevation of V*b equivalent to that observed during anaemia, but in the absence of lowered haematocrit, increased PaCO(2) significantly by 1.49+/-0.74 to 1.64+/-0.78 torr (N=5) without affecting PaO(2). These findings suggest that CO(2) excretion in bullhead, as in rainbow trout, is effectively diffusion-limited, and that approximately half of the increase in PaCO(2) measured during the initial 2 h of anaemia results from the impact of increased blood flow (hence decreased gill transit time) in a diffusion-limited system.     

The development and role of international biological reference materials in the diagnosis of anaemia

Anaemia is a major global health problem. Although the main cause is iron deficiency, anaemia also results from other nutritional deficiencies (folate and vitamin B₁₂), haemolytic disorders including haemoglobinopathies, and bone marrow disorders. Accurate diagnosis of anaemia is dependent on reliable diagnostic tests and reference ranges, which in turn are dependent on effective standardisation. Standardisation is achieved through the availability of reference materials and reference measurement procedures. International biological reference materials have therefore been developed to standardise and control diagnostic tests for anaemia for a diverse range of analytes including total haemoglobin and haemoglobin types, ferritin, the serum transferrin receptor, serum vitamin B₁₂ and folate, whole blood folate, and alloantibodies which mediate immune haemolytic anaemia.