British Journal of Haematology

Dixon, Richard H.; Rosse, Wendell F.

doi: 10.1111/j.1365-2141.1975.tb00843.xpmid: 1103953

Grasso, J. A.; Sullivan, A. L.; Sullivan, L. W.

doi: 10.1111/j.1365-2141.1975.tb00844.xpmid: 1201233

Summary. Marrow aspirates from sickle cell patients were examined without prior deoxygenation and revealed the presence of a variable proportion (10–30%) of sickled red cells and reticulocytes. The main feature of sickled red cells was the presence of 17.6 nm fibres arranged in hexagonal order, and to a lesser extent in square or rectangular array, to form bundles of varying size and compactness which occupied the entire cytoplasm. The sickling pattern in reticulocytes was more variable. Some reticulocytes contained highly‐ordered bundles of 17.6 nm fibres whose structure and distribution was identical to that in red cells from which they could be distinguished only by their content of organelles. Many reticulocytes exhibited less‐organized fibre patterns ranging from localized aggregates to poorly‐ordered regions of short fibres and filaments lacking apparent preferential orientation. In these cells, the bulk of the cytoplasm was not polymerized. Haemoglobin polymerization in reticulocytes led to entrapment of ribosomes in concentrated foci among the fibres. Frequently, iron‐laden mitochondria were associated with sickled reticulocytes. The variation in pattern of sickling seen in reticulocytes is attributed to possible differences in concentration of Hb S. Correlative studies have shown that fibres were not induced by fixation procedures. Marked phagocytic degradation of sickled cells by macrophages was observed. The results are interpreted to indicate the possible intramedullary phagocytosis of red cells and reticulocytes, predisposed to sickling in the marrow. However, the data are also consistent with the removal of sickled elements from the circulating blood.

Walter, Harry; Krob, Eugene J.; Ascher, Galli S.

doi: 10.1111/j.1365-2141.1975.tb00845.xpmid: 1201234

Summary. Partition of cells in two‐polymer aqueous phases is an extremely sensitive method for the separation of cells and for tracing subtle changes in the cells' membrane surface properties (primarily surface charge) as a function of in vivo processes (e.g. differentiation, maturation, ageing). Combining isotope labelling and cell countercurrent distribution techniques we have examined the membrane surface properties of rat reticulocytes produced in response to erythroid stress by bleeding. It was found that the rapid increase in the partition coefficient of normal reticulocytes subsequent to release into the peripheral blood (which reflects a rapid increase in membrane surface charge) is absent in reticulocytes produced in response to bleeding. In this way the reticulocytes behave like those produced in response to repeated phenylhydrazine injections. ‘Stress reticulocytes’ never mature to erythrocytes having normal membrane surface properties as judged by partition. The experiments show that our previous results are not due to phenylhydrazine per se but rather to the production of ‘stress reticulocytes’. Further, if remodelling of the phlebotomy‐induced reticulocyte cell membrane occurs in the circulation as has been suggested it does not lead to a cell with normal membrane surface properties. Whether the abnormal membrane surface properties of ‘stress reticulocytes’ and of the erythrocytes to which they give rise affect the cells' life‐span is not clear since phenylhydrazine‐induced ‘stress reticulocytes’ have been reported to have at least a component of short‐lived cells while the phlebotomy‐induced cells are said to have a virtually normal life‐span. It is likely that reticulocytes produced in severe anaemias, in general, mature to erythrocytes having abnormal surface properties (i.e. lower membrane surface charge).

Sansone, G.; Perroni, L.; Yoshida, A.

doi: 10.1111/j.1365-2141.1975.tb00846.xpmid: 1201235

Summary. Screening for the G6PD deficiency was carried out at the Maternity Division of the Galliera Hospital in Genoa, Italy. Two groups of subjects with hyperbilirubinaemia of non‐immunological origin were examined: (a) 302 newborn babies of Sardinian extraction (on cord blood) and (b) 201 newborn babies of south Italian ancestry (on peripheral blood). Among 503 subjects, 43 showed an enzyme deficiency; in 39 the defect was of the Mediterranean type. In one case, previously described, the enzyme was of the A type. In the remaining cases three different variants were identified. In the present work these three cases, each with severe neonatal jaundice, are reported. Their parents originated from Calabria, from Sardinia and from Sicily. The abnormal enzymes are respectively designated as GdDebrousse‐like, GdGallura and GdAgrigento.

Beard, M. E. J.; Bateman, C. J. T.; Crowther, D. C.; Wrigley, P. F. M.; Whitehouse, J. M. A.; Fairley, G. Hamilton; Scott, Ronald Bodley

doi: 10.1111/j.1365-2141.1975.tb00847.xpmid: 1059476

Summary. The clinical course, diagnosis and management of nine cases of hypoplastic acute myelogenous leukaemia are described. Such cases may follow a slowly progressive course and should not receive anti‐leukaemic chemotherapy unless the disease is advancing rapidly or unless some specific complication develops. If chemotherapy has to be given, usually because of severe and recurrent infections, then prompt and prolonged remission of disease may occur.

Schiffer, Charles A.; Levi, John A.; Wiernik, Peter H.

doi: 10.1111/j.1365-2141.1975.tb00848.xpmid: 1201236

Summary. Buffy coat preparations of peripheral blood from two patients with Stage IVB Hodgkin's disease, 33 patients with Hodgkin's disease undergoing complete staging including laparotomy, 12 normal controls and eight patients with viral upper respiratory infections were examined for the presence of abnormal cells. Primitive histiocytic cells were seen in the two patients with IVB disease and in two patients with IIIB disease all of whom proved to be refractory to chemotherapy. Reed‐Sternberg cells were seen in one IVB patient. Large cells with basophilic, agranular cytoplasm and moderately convoluted nuclei often with a perinuclear halo and nucleoli were detected in 30 of 35 patients with Hodgkin's disease, five of eight patients with viral infections but in none of 12 normal controls. Other abnormal large cells with round nuclei, occasional prominent nucleoli, grey to lightly eosinophilic cytoplasm and diffuse granulation were seen almost exclusively in Hodgkin's disease. There was no correlation between histology, stage, splenic involvement, skin test positivity and response to therapy with the presence or frequency of either of these cell types. This observation suggests that these cells are more likely to be ‘reactive’rather than malignant. It is unusual for abnormal histiocytic cells to circulate in Hodgkin's disease and their presence may be a poor prognostic sign. The presence of other types of large atypical cells is not indicative of either haematogenous spread of Hodgkin's disease or of poor prognosis after treatment with radiotherapy alone.

McGuffin, Robert; Goff, Paul; Hillman, R. S.

doi: 10.1111/j.1365-2141.1975.tb00849.xpmid: 1201237

Summary. Studies of the rate of depletion of serum and tissue methylated and nonmethylated folates were carried out in rats maintained for long periods on either a folate deficient (sucrose‐water/sulphathiazole) diet or a deficient diet plus high alcohol intake. By means of implantation of a feeding gastrostomy tube, it was possible to sustain constant blood ethanol levels of between 50 and 300 mg/dl for 3–4 weeks with relatively normal calorie intake and without death of the animal. Using this animal model, which closely resembles severe alcoholism in man, a very rapid depression in serum 5‐methyl tetrahydrofolate was observed similar to that reported in alcoholic man. At the same time, release of folates from liver stores was unimpaired by alcohol ingestion. Liver folate store depletion rates were identical for alcoholic and folate starved animals. The explanation for the sudden alcohol suppression of serum folate levels must, therefore, be sought at a point in the internal metabolic sequences of folate other than the delivery of folate stores to plasma.

Tsan, Min‐Fu; McIntyre, Patricia A.

doi: 10.1111/j.1365-2141.1975.tb00850.xpmid: 1201238

Summary. The effect of propylthiouracil on glucose metabolism in human polymorphonuclear leucocytes was studied. At a therapeutically achievable concentration (0.1 mm), propylthiouracil stimulated hexose monophosphate shunt activity in normal leucocytes during phagocytosis but not in resting cells. However, in the presence of hydrogen peroxide it stimulated hexose monophosphate shunt activity in resting cells, and in the soluble fraction when reduced glutathione and reduced nictotinamide adenine dinucleotide phosphate (NADPH) were also present. Propylthiouracil had no effect on glucose‐1‐C oxidation in either phagocytosing or resting leucocytes obtained from two male patients with chronic granulomatous disease. Stimulation of the hexose monophosphate shunt activity in normal leucocytes during phagocytosis also was demonstrated with methimazole, thiouracil and thiourea, but not with adenine, uracil or urea. There was an apparent minimal common structure requirement in thiourea. Propylthiouracil had no effect on phagocytosis, formate oxidation, glucose‐6‐phosphate dehydrogenase, 6‐phosphogluconate dehydrogenase or catalase activities. Thus, the stimulation of the hexose monophosphate shunt activity by propylthiouracil is dependent on hydrogen peroxide and is best explained by its stimulation or participation in the glutathione cycle.

Dallman, P. R.; Siimes, M. A.; Manies, E. C.

doi: 10.1111/j.1365-2141.1975.tb00851.xpmid: 1201239

Summary. This study was designed to determine the content of non‐haem iron in the brain as iron deficiency develops in the rapidly growing rat. Rats were provided with either an iron‐deficient diet or an identical control diet with added ferrous sulphate starting at 10 d of age and continuing after weaning at 21 d. At 28 or 48 d of age the deficient animals received 5 mg of iron (iron dextran) i.m. and were placed on the control diet regimen. The deficient animals had a concentration of non‐haem iron in the brain that was 27% below the control value at 28 d and 22% below at 48 d. After 14–45 d of iron treatment, the non‐haem iron remained depressed, 19–29% below the control means (P<0.05 to 0.001). Ferritin iron in brain also remained depressed, 33–42% below the control means (P<0.01). In contrast, haematocrit, liver non‐haem iron, and liver ferritin iron, although they were more profoundly depressed in the iron‐deficient animals, promptly returned to control values after treatment with iron. Thus, a brief period of severe iron deficiency in the young rat resulted in a deficit of brain iron that persisted in the adult animal despite an adequate intake of iron.

Gallimore, M. J.

doi: 10.1111/j.1365-2141.1975.tb00852.xpmid: 53065

Summary. Four protease inhibitors have been identified in human serum and methods for their isolation are described. After removal of the euglobulin fraction, serum was submitted to ion exchange chromatography and gel filtration, and fractions were tested for inhibition of the lysis of plasminogen‐deficient fibrin clots by plasmin, trypsin and elastase. In addition inhibitors of plasminogen activation were sought by studying the effects of separated fractions on the lysis of plasminogenrich fibrin clots by urokinase. Examination by immunophoresis showed that three of the separated inhibitors were α2‐macroglobulin, α1‐antitrypsin and inter‐α‐trypsin inhibitor. The fourth antiprotease was a powerful inhibitor of both urokinase‐induced and plasmin‐induced clot lysis, and was identified as an inter‐α‐globulin from its electrophoretic mobility in agarose gels.