Hereditary ovalocytosis is common in some areas of Melanesia and South East Asia where malaria is endemic. These red cells resist invasion by malarial parasites in vitro and ovalocytic individuals are less parasitized than normal. This has been attributed to the greater rigidity of ovalocytic red cells. It has been suggested that South East Asian ovalocytosis results from the heterozygous presence of an altered membrane anion transporter (band 3). We have used the polymerase chain reaction to clone the abnormal band 3 complementary DNA from an ovalocytic of Indian origin and found two changes from the normal protein: a point mutation (Lys 56----Glu) and the deletion of the sequence AFSPQVLAA (residues 400-408), but no evidence for an N-terminal extension. The deletion is also found in the abnormal band 3 of South East Asian ovalocytes and seems to be responsible for the unusual properties of the ovalocytic red cell. We show here that the membrane domain of the abnormal ovalocyte band 3 has a substantially altered structure and that the protein is defective in anion transport activity. The changed transport properties of the red cells may have a role in the reduced parasitaemia of ovalocytic individuals.

The effect of dipotassium ethylenediaminetetra-acetic acid (EDTA) on platelet count and mean volume (MPV) was evaluated using two routine measurement systems, a Coulter S Plus (Phase 1) (S+) and a Technicon H6000 (H6000). In normal subjects (n = 29) MPV increased by 17% during 39 h storage in EDTA when measured by the S+. In contrast MPV decreased by 22% when measured by the H6000. MPV differences of up to 40% were observed between the two systems. Concomitant platelet counts, in both systems, changed by less than 4%. A mathematical model of the variation of MPV with storage time was constructed, enabling experimental results to be extrapolated, with accuracy, to time zero (MPV0). The H6000 average MPV0 was significantly larger than the S+ average MPV0. Using the anticoagulant sodium citrate and prostaglandin E1 (NaCitrate-PGE1) there were no significant changes in MPV measured by the S+ during 7 h storage, although a linear decrease in platelet count was observed. A decrease in H6000 MPV was observed whether the blood was stored in EDTA or NaCitrate-PGE1. Methodology, anti-coagulation and storage time all influence MPV. Until these determinants are standardised the clinical value of MPV cannot be assessed.

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South-east Asian ovalocytosis (SAO) results from the heterozygous presence of an abnormal band 3, which causes several alterations in the properties of the erythrocytes. Although earlier studies suggested that SAO erythrocytes are refractory to invasion in vitro by the malarial parasite Plasmodium falciparum, a more recent study showed that fresh SAO cells were invaded by the parasites, but became resistant to invasion on storage because intracellular ATP was depleted more rapidly than normal. Here we show that SAO red cells are much more leaky to sodium and potassium than normal red cells when stored in the cold. This leak was much less marked when the cells were stored at 25 or 37 degreesC. Incubation for 3.5 h at 37 degreesC of cold-stored SAO red cells did not restore sodium and potassium to normal levels, probably because the depleted ATP level in cold-stored SAO red cells is further reduced with incubation at 37 degreesC. The increased leakiness of SAO red cells is non-specific and extends to calcium ions, taurine, mannitol and sucrose. These results suggest that SAO red cells undergo a structural change on cooling. Since many of the reports describing altered properties of SAO red cells have used cells which have been stored in the cold, these results need re-evaluation using never-chilled SAO red cells to assess whether the cells have the same abnormal properties under in vivo conditions.

The performance of the Coulter STKS (Coulter, Hialeah, FL) was evaluated in a busy computerized teaching hospital laboratory. The STKS was compared with a Coulter S Plus IV and manually performed 400 white blood-cell differentials. The measured blood-count parameters (i.e., white blood cells [WBCs], red blood cells [RBCs], hemoglobulin [Hb], mean corpuscular volume [MCV], and platelets [PLTs]), compared very well between the two aperture impedance-based systems; precision, linearity, and lack of carryover were excellent. The STKS WBC differential (DIFF), derived from a combination of aperture impedance, aperture conductance, and laser light scatter, also was precise; linear and carryover were insignificant. The DIFFs (n = 424) compared well to the manual WBC differentials, with r values of 0.97, 0.97, 0.73, and 0.86 for neutrophils, lymphocytes, monocytes, and eosinophils, respectively. The DIFF and Suspect Flagging system produced 6.2% false negatives and 2.6% false positives when compared with the manual technique. These were further investigated and discussed. STKS DIFFs were stable for 18 to 24 hours in normal samples anticoagulated with K2EDTA and stored at 20 degrees C prior to analysis. Storage in the same anticoagulant at 4 degrees C and immediate aspiration preserved the DIFF analysis for considerably longer than 24 hours. These performance characteristics make the STKS a significant advancement in automated hematology.