bluuebunny/crossref_metadata_2025 huggingface dataset:
Red‐Cell Abnormalities in HEMPAS (Hereditary Erythroblastic Multinuclearity with a Positive Acidified‐Serum Test)
Authors: Crookston J. H. , Crookston Marie C. , Rosse W. F.
Published: March 2008

Summary. Abnormalities of the membrane of HEMPAS red cells have been demonstrated by certain serological tests and in freeze‐etch electron micrographs. There is also a marked increase in the activity of certain red‐cell enzymes.

The positive acidified‐serum lysis test has proved useful in distinguishing HEMPAS from other kinds of hereditary dyserythropoietic anaemia. The greatly increased reactivity with anti‐i (although characteristic of HEMPAS) is a less specific finding, since enhancement of the i antigen is found in other dyserythropoietic anaemias, hereditary and acquired. Although the lysis of HEMPAS cells by acidified serum at first suggested that their membrane defect was similar to that present in PNH, further investigation showed that the mechanism of activation of complement is different in the two conditions. Lysis of HEMPAS cells depends on the binding of complement by a naturally‐occurring, cold antibody (“anti‐HEMPAS”) specific for an antigen present on HEMPAS cells, but not detectable on PNH or normal cells. Unlike PNH cells, HEMPAS cells are not lysed by complement in the absence of antibody.

As judged by tests with anti‐I and human complement, PNH cells have a true increase in sensitivity to complement in that the binding of only a small number of complement sequences is sufficient to effect lysis. The increased lysis of HEMPAS cells by anti‐I appears to be due to the binding by antibody of many more complement sequences than are bound to PNH or normal cells. With some examples of anti‐I, an increased uptake of antibody by HEMPAS cells also contributes to their increased lysis.
Iron Absorption in Indians Studied, by Whole Body Counting: a Comparison of Iron Compounds Used in Salt Fortification
Authors: Narasinga Rao B. S. , Prasad Surendra , Apte Sharad V.
Published: March 2008

Summary

Fortification of common salt (NaCl) with iron has been suggested, for preventing iron deficiency anaemia in India. From a technological point of view three insoluble iron compounds: sodium iron pyrophosphate, ferric orthophosphate and ferric pyrophosphate have been considered suitable for this purpose. Absorption of iron from these compounds and ferrous sulphate was studied in 45 healthy male adult subjects employing 59Fe labelled salts and whole body counting. “When given alone, mean absorption of iron from ferric orthophosphate, sodium iron pryophos‐phate and ferric pyrophosphate were 8.3%, 6.3% and 0% respectively, while absorption from ferrous ascorbate was 30.6%. However, when administered with a meal of the type normally consumed in India, iron absorption from sodium iron pyrophosphate was unacceptably low, while that from ferrous sulphate was only 3.7%. It is concluded that these insoluble iron compounds are not acceptable for fortification of common salt from the point of view of availability of iron.
A Study of Storage Iron in the Placental and Foetal Tissues of the Rat
Authors: Kaufman Nathan , Wyllie John C.
Published: March 2008

Summary

Part of the iron entering the placenta late in gestation is stored as ferri‐tin and haemosiderin; the function of placental storage iron is not known. 50–60% of foetal iron is found within storage compounds. There are relatively few studies of the foetal tissues responsible for the synthesis of storage iron‐protein.

In order to investigate these aspects of iron metabolism we measured the quantity and concentration of ferritin and haemosiderin iron in the placental and foetal tissues of the rat during the latter part of gestation; the incorporation of 59Fe into these compounds was also investigated.

Placental ferritin and haemosiderin iron reached peak values on day 17. A 40% decrease in the quantity of ferritin iron by day 19, without a corresponding change in placental weight, indicated mobilization of iron from placental ferritin; haemosiderin iron remained unchanged. On day 21 (parturition) both iron storage compounds again increased in quantity. The amounts of storage iron deposited in the allantoic placenta and yolk sac were similar until day 21 when the former tissue contained 75% of the total quantity. The concentration of ferritin iron was higher in the yolk sac on days 17 and 19.

The major portion of foetal ferritin and haemosiderin was deposited late in gestation. The carcass and liver were the principal sites involved although small amounts were found in the spleen and gastrointestinal tract. The incorporation of 59Fe by ferritin and haemosiderin confirmed the results obtained by chemical analysis and also showed that there was a steady addition of iron to both compounds throughout the latter part of gestation.
Two Progressive Inhibitors of Factor Xa in Human Blood
Authors: Marciniak Ewa , Tsukamura Sumiko
Published: March 2008

Summary

The limited ability of human blood to inactivate human factor Xa was found to be markedly increased in human plasma and serum intensively absorbed by aluminium hydroxide. Using gel‐filtration analysis on Sephadex G‐200 it was demonstrated that the activity brought about by adsorption resides in the macro‐globulin fraction. This fraction, as obtained from nonabsorbed plasma or serum, has low antifactor‐Xa activity; it represents a progressive inhibitor of this enzyme, additional to antithrombin III. The increase of inhibitory activity due to adsorption is probably based on removal of a substance associated with macroglobulin in vivo, and on uncovering new binding sites for factor Xa. Whereas antithrombin III and the antifactor Xa of lower molecular size are probably represented by a single protease inhibitor which may be potentiated by heparin, the macroglobular anti‐factor Xa is not closely related to antithrombin activity and is unaffected by heparin.
The Role of Erythropoietin in Prenatal Erythropoiesis of Congenitally Anaemic Flexed‐tailed (f/f) Mice
Authors: Bateman A. E. , Cole R. J. , Regan T. , Tarbutt R. G.
Published: March 2008

Summary

The effects of the f gene on prenatal changes in liver cellularity, erythroid cell maturation, RNA, DNA, and haemoglobin synthesis, and sensitivity to erythropoietin have been investigated, f/f foetuses are smaller than f/+ but still develop a progressive deficiency in haemoglobin content/unit weight of tissue, which is 40% of normal at birth. Early f/f livers contain fewer erythroblasts, maximum numbers are reached later, and changes in relative proportions of early and late erythroblasts are retarded. Erythroid tissue and erythropoietin responsive cells persist longer in f/f than in f/+ livers. Erythropoietin induced RNA and DNA synthesis is higher in f/f than in f/+ foetal liver cell cultures. Erythropoietin causes multiphasic increments in haem synthesis in vitro. Maximum levels of synthesis by 13 day liver cells are reached later f/f but over 36 hr similar amounts are accumulated. Erythropoietin maintains the presence of early erythroid cells in both f/f and f/+ foetal liver cultures, and enhances the production of polychromatic cells.

Circulating f/f foetal reticulocytes have a reduced capacity to utilize exogenous iron for haem synthesis. The deficient haemoglobin content of f/f foetuses could be caused by this abnormality and by reductions in erythroid precursor cell numbers. The reticulocyte abnormality may be a direct or indirect consequence of a failure of erythroid colonization of the foetal liver, and of the effects of secondary compensation for this failure.
Autoantibodies of U Blood Group Specificity in Autoimmune Haemolytic Anaemia
Authors: Marsh W. L. , Reid Mawon E. , Scott E. Patricia
Published: March 2008

Summary

Autoantibody specificity in acquired haemolytic anaemia of the warm antibody type has been thought to be directed against Rh antigens or the precursors from which they arise. In many cases Rhnull red cells fail to react with these antibodies, providing apparent support for the assumption that they have Rh specificity. Rhnull red cells are also non‐reactive with anti‐U sera, and in addition have abnormal S and s antigens. We have studied 50 cases of warm antibody autoimmune haemolytic anaemia looking particularly for possible U, S or s blood group specificity of the causative antibodies. In the majority of cases the antibodies were found to be directed against antigens of the Rh complex, but in three cases multi‐specific autoantibodies were shown to have complex specificity involving the Rh and U blood groups.
Defective Erythrocyte Pyruvate Kinase with Impaired Kinetics and Reduced Optimal Activity
Authors: Paglia Donald E. , Valentine William N. , Rucknagel Donald L. , Wittenberg Julie
Published: March 2008

Summary

A unique mutant form of erythrocyte pyruvate kinase has been found associated with chronic haemolytic anaemia in a child who is apparently doubly heterozygous for the mutant isoenzyme and for pyruvate kinase deficiency of the classical quantitative type. Clinical and laboratory findings conformed closely to those typically observed in homozygous pyruvate kinase deficiency anaemia. Assayed in fresh haemolysates, the isoenzyme exhibited reduced optimal activity (c 45% of normal) and an increased Michaelis constant for phosphoenolpyruvate (four to five times greater than normal). The kinetic anomaly was only partially corrected by activation with fructose‐1,6‐disphosphate. Despite some common characteristics, this isoenzyme appears distinct from others reported in the literature and lends support to the polymorphous nature of heritable baemolytic anaemias secondary to defective pyruvate kinase.
δβ‐Thalassaemia in a Chinese Family
Authors: Mann J. R. , MacNeish A. S. , Bannister D. , Clegg J. B. , Wood W. G. , Weatherall D. J.
Published: March 2008

Summary. δβ‐Thalassaemia has been observed for the first time in individuals of Chinese origin. The clinical and haematological features have been characterized in the heterozygous state and in the double heterozygous state withβ‐thalassaemia. Studies of haemoglobin synthesis indicate that the degree of globin chain imbalance in β‐thalassaemia is less than that found in β‐thalassaemia. Analysis of the foetal haemoglobin has shown that all individuals carrying the δβ‐thalassaemia gene in this family synthesize only the α2γ2136 glycine variety. This type of foetal haemoglobin has been found previously only in two Negro individuals heterozygous for hereditary persistence of foetal haemoglobin (HPFH). The genetic relationships between δβ‐thalassaemia and HPFH are discussed in the light of these new findings.
Diaphorase Activity and Variants in Normal Adults and Newborns
Authors: Eng Lie‐Injo Luan , Loo May , Fah Foo Kon
Published: March 2008

Summary. Red cell NADH‐dependent methaemoglobin reductase (diaphorase) activity was estimated in 126 normal healthy adults and 556 newborns of the three major racial groups in Malaysia. The mean enzyme activity in adults was 3.34 units/mg Hb (SD ± 0.63) and in newborns 1.75 u/mg Hb (SD ± 0.48). The difference between the two means is statistically highly significant. Indians, both adults and infants had the lowest mean enzyme activity. Mean activity in a group of newborns whose birth weights were 2.7 kg or more was significantly higher than in a group with lower birth weights. Sixteen newborns had enzyme activity of less than 1 u/mg Hb. Two of them each had one parent whose enzyme level was also very low. Both parents of three others had normal enzyme activity. The lower limit of normal in newborns seemed to overlap with that found in subjects with hereditary methaemoglobin reductase deficiency. One newborn with an extremely low enzyme level had methaemoglobinaemia, but its genetic nature could not be proved because the parents were not available for study. Examination of 108 adults and 527 newborns for diaphorase electrophoretic variants revealed two different and unusual phenotypes in a few Chinese and Indians. Family studies showed that they are genetically determined.
Apparent Paradoxical Patterns of Anaerobic Glycolysis and Hexokinase Activity in the Red Blood Cells of Acutely Bled Rats, with Evidence that the Responses were in Part Hormone‐Dependent
Authors: Sabine Jean C.
Published: March 2008

Summary. Rats were bled of c 25% of their blood volume, and anaerobic glycolysis (AG) and hexokinase activity (HxK) were assayed in the red blood cells. After 2–14 days the animals were sacrificed and AG and HxK again assayed. Four different responses were observed: (1) both parameters were elevated following blood loss in only four out of 87 intact males, as expected if increased proportions of normal young red cells are present; (2) both parameters were decreased in six animals. In the majority, two paradoxical patterns were found; (3) increased AG with decreased HxK; and (4) decreased AG with increased HxK. Response 3 was found only in immature animals, and Response 4 in animals at or beyond the minimal breeding age of 12 weeks. Results in castrated males and adult females were indistinguishable from each other and from those in immature males. In testosterone‐treated females the response was modified toward but not converted to Response 4.

It is proposed that Response 3 was probably independent of sex hormones. but was converted to Response 4 when a full complement of male hormones was present. Whatever the underlying mechanism, the finding of four distinct respenses indicates that, in terms of the two parameters assayed in this investigation, there was no entity which could be termed a ‘stress reticulocyte’. There is a strong likelihood that factors not detectable by the methods used were responsible for the paradoxical findings.
Iron Deficiency and Dyserythropoiesis
Authors: Hill R. S. , Pettit J. E. , Tttersall M. H. N. , Kiley N. , Lewis S. M.
Published: March 2008

Summary. In 10 patients with Fe deficiency significant dyserythropoiesis was found. The degree of dyserythropoiesis provided a reliable indication of the severity of the iron deficiency as measured by the serum iron and total iron binding capacity; a positive correlation (P < 0.002.) being found between the % incidence of dyserythropoiesis in the bone marrow and both the serum iron and % iron saturation. The frequent occurrence of basophilic stippling in the erythroblasts of these patients suggested that the presence or absence of stippling does not help differentiate between iron deficiency and other causes of hypochromic anaemia.
Porphyrin Metabolism in Experimental Hepatic Siderosis in the Rat
Authors: Taljaard J. J. F. , Shanley B. C. , Deppe W. M. , Joubert S. M.
Published: March 2008

Summary. Rats previously rendered siderotic by intraperitoneal administration of iron‐dextran became porphyric much more rapidly than non‐siderotic animals when fed hexachlorobenzene. Fluorescence microscopy showed that initially porphyrin storage was focal and confined to the centrilobular zone but later became diffuse throughout the lobule. Analysis of liver extracts by thin layer chromatography revealed only 8‐ and 7‐carboxyl porphyrins which also predominated in the urine. The patterns resembled those seen in patients with symptomatic porphyria. These data are interpreted as further evidence that siderosis is an important factor in the derangement of hepatic haem biosynthesis which leads to the development of symptomatic porphyria.
Platelet Coagulant Activities in Thrombasthenia
Authors: Walsh P. N.
Published: March 2008

Summary. Methods for assaying four platelet coagulant activities are presented. The results of performing these assays in six patients with thrombasthenia revealed decreased contact‐forming activity (CPFA) and an absence of platelet response to adenosine diphosphate in the CPFA assay. All patients had normal or raised intrinsic factor‐Xa forming activity (XaFA) and decreased platelet factor‐3 activity (PF3A), the level of which related poorly to bleeding history. The four patients most severely affected clinically lacked collagen‐induced coagulant activity (CICA), whereas CICA was present in the other two. The important determinants of bleeding severity in thrombasthenia may be the platelet coagulant activities concerned with the initiation of intrinsic clotting (i.e. CICA and CPFA) and not those concerned with the subsequent interaction of clotting factors other than the contact factors (i.e. XaFA and PF3A). It is concluded that each of the four coagulant activities studied is separate and distinct from the others, and each has its own biochemical determinants.
Porphyrin Metabolism in Experimental Hepatic Siderosis in the Rat
Authors: Taljaard J. J. F. , Shanley B. C. , Deppe W. M. , Joubert S. M.
Published: March 2008

Summary. The combination of iron overload and hexachlorobenzene (HCB) feeding in rats produces an ‘experimental symptomatic porphyria’. Studies of hepatic haem biosynthesis in this model showed a decrease in total liver haem and cytochromes P450 and b5, together with decreased incorporation of 14C‐succinate into liver haem and an absence of uroporphyrinogen decarboxylase activity. A marked alteration was found in the redox state of the liver cell cytoplasm both in siderotic and non‐siderotic animals after HCB feeding; the ratio NAD: NADH was increased more than two‐fold. These findings suggest that the primary effect of HCB with regard to liver haem biosynthesis may be this change in the redox state, possibly leading to decreased ability to maintain porphyrinogens in the reduced form and that the additional effect of iron overload is mediated by impairment of uroporphyrinogen decarboxylase activity and an increased rate of degradation of microsomal drug detoxifying cytochromes. It is probable that in human symptomatic porphyria a number of factors are similarly responsible for the excessive production and excretion of porphyrins.
A Simple Method for the Detection of Unstable Haemoglobins
Authors: Carrell R. W. , Kay R.
Published: March 2008

Summary. A simple test is described for the detection of unstable haemoglobins by the addition of haemolysate to an isopropanol/buffer solution at 37°C. Over 200 normal haemolysates have been tested giving no precipitation up to 30 min incubation whilst six samples containing an unstable haemoglobin gave a precipitate within 5 min, which was flocculent by 20 min. As well as being a simple and effective screening test, the procedure is suitable for the purification of unstable haemoglobins for further studies but suffers from limitations, probably in common with the heat precipitation method, in the quantitation of the abnormal haemoglobin. Its use as a research tool is illustrated by comparative measurements of the stabilities of various forms of haemoglobin. These show deoxyhaemoglobin to be considerably more stable than oxyhaemoglobin thus supporting suggested interchain bondings in deoxyhaemoglobin.
Red Cell Survival Studies in Haemoglobin H Disease using [51Cr] Chromate and [32P]Di‐isopropyl Phosphofluoridate
Authors: Tso S. C.
Published: March 2008

Summary. Red‐cell survival was studied in eight patients with haemoglobin H disease in the steady state, using simultaneously [51Cr]chromate and [32P]di‐iso‐propyl phosphofluoridate (DF32P). A shortened survival apparently due to a process of random destruction was observed in all patients, and the rate of 51Cr elution calculated from the DF32P data was within the normal range. In‐vitro studies showed that most of the DF32P label was intraerythrocytic, bound to the erythrocyte B esterase and not to haemoglobin. In view of these findings, it is suggested that [51Cr]chromate is a valid label for studying red‐cell survival in haemoglobin H disease.
Study of a Granulopoietin from Endotoxin‐Stimulated Mouse Plasma
Authors: Bierman Howard R. , Hood James E.
Published: March 2008

Summary: The presence of a granulopoietic stimulating factor was demonstrated in the plasma and marrow interstitial fluid of mice 18 hr following the injection of E. coli endotoxin or turpentine, when maximum polymorphonuclear leucocyte depletion occurs in the marrow. Stimulation was evident in the marrows of C57B1 mice from an increase in the absolute number of immature granulocytes, as well as by an increased 3H‐thymidine uptake in the marrow. The effect was also shown to be concentration‐dependent. It can be differentiated from the stimulatory effect which results from marrow granulocyte depletion (a secondary effect), since there was no significant loss of granulocytes from the marrow upon administration of the active preparations. It is therefore distinct from Gordon's leucocyte inducing factor.
Monocyte Kinetic Studies in Normal and Disease States
Authors: Meuret G. , Hoffmann G.
Published: March 2008

Summary. Monocyte kinetics were studied in eight normal subjects and 27 patients suffering from various diseases: acute, subacute and chronic infections, malignant tumours, Boeck's sarcoid, neutropenia, and diseases associated with chronic exanthemata or splenomegaly. Autotransfusion of blood cells labelled with 3H‐diisopropylfluorophosphate in vitro was carried out and the fate of these cells in vivo was followed by autoradiographs of leucocyte concentrates prepared from venous samples.

Within a few minutes the transfused monocytes equilibrated with the total blood monocyte pool (TBMP). As TBMP proved to be larger than the circulating monocyte pool (CMP), the existence of a marginal monocyte pool (MMP) was postulated. In normal subjects the CMP:MMP ratio averaged 1:3.5. Moderate deviations occurred in disease states. Monocytes left the vascular system at an exponential rate, the mean normal T4 being 8.4 hr. Slight prolongation of T4 was observed in some patients with monocytosis, the maximal values reaching 15 hr. Shortened T4 was found in one patient with acute infection (4.0 hr) and in another with gross splenomegaly (3.5 hr). The monocyte turnover rate (MTR) in normal subjects averaged 6 × 108 monocytes/hr or 7 × 106 monocytes/hr/kg. Highly significant positive correlations were evaluated between the blood monocyte count and TBMP or MTR respectively.
A Quantitative Evaluation of the Inhibition of Platelet Aggregation by Low Molecular Weight Degradation Products of Fibrinogen
Authors: Solum Nils Olav , Rigollot Colette , Budzynski Andrzej Z. , Marder Victor J.
Published: March 2008

Summary. The inhibitory effect of low molecular weight degradation products of fibrinogen (peptides) on in vitro platelet aggregation has been studied quantitatively. In our test system (0.3 μm ADP at 25°C) only concentrations higher than 0.65 mg/ml showed significant inhibitory activity. The higher concentrations of peptides also inhibited platelet aggregation induced by adrenaline, collagen and thrombin. Ultrafiltrates of plasma from patients receiving streptokinase showed no increase in inhibition of platelet aggregation as compared with pretreatment samples. This supports the idea that the peptides produced by streptokinase‐induced fibrinogcnolysis exert a quantitatively weak effect. Peptides produced by digestion of serum with trypsin showed inhibitory activity similar to that of the peptides produced by digestion of fibrinogen with plasmin, indicating that the inhibitory effect is unspccific as far as the protein source and the acting enzyme are concerned. Some specificity may exist, however, related to size and/or charge of the peptides, as suggested by the present experiments. These data do not support the view that the low molecular weight degradation products of fibrinogen significantly impair platelet aggregation in the fibrinolytic states.
Effect of Alpha‐ and Beta‐blocking Drugs on the Clotting and Fibrinolytic Response to Venous Stasis in Man
Authors: Ponari O. , Civardi E. , Megha A. , Pini M. , Potì R. , Dettori A. G.
Published: March 2008

Summary. The changes in coagulability and in fibrinolytic activity induced in local blood by venous stasis in man are not prevented by blocking of alpha‐ and beta‐adrenergic receptors.

Administration of l‐propranolol and of l‐propranolol plus phentolamine in adequate doses just before the execution of a tourniquet test failed to inhibit the major changes, i.e. rise in factor‐VIII level, shortening of partial thromboplastin time (PTT) and euglobulin lysis time (ELT), hypercoagulable pattern on thromboelastogram, following venous stasis. Some other clotting tests remained unchanged in all experimental conditions.

Physiopathological implications of these results are discussed, taking into account the possible role of the adrenergic system in the control of a postulated equilibrium between blood coagulation and fibrinolysis.
Erythrocyte Production in Mice Infected by the Polycythaemia‐Inducing Friend Virus or by the Anaemia‐Inducing Friend Virus
Authors: Tambourin P. E. , Gallien‐Lartigue O. , Wendling F. , Huaulme D.
Published: March 2008

Summary. The changes induced by infection with the anaemia‐inducing (FVA) or the polycythaemia‐inducing (FVP) strain of Friend virus on PCV, RBC, reticulocyte counts, red‐cell and plasma volume and reticulocyte maturation time were studied in Swiss mice for 7 weeks after viral infection. The data indicated that the increase in total blood volume is mainly due to the augmentation of the spleen and liver blood volume in FVA‐disease, whereas it results from the large numbers of red cells produced by the spleen in FVP‐disease.

The daily output of reticulocytes was increased four‐fold in FVA infected mice, and thirty‐fold in FVP infected mice by the 50th day post‐infection. The total redcell production for the duration of the experiment was 3 times normal for the FVA group and 13 times normal for the FVP group. A theoretical estimate of the mean red‐cell lifespan in infected mice gave 8 1/2 days for the FVP‐disease and 22 days for the FVA‐disease as opposed to 40 days for the normal mouse.
Evidence for a Humoral Agent Capable of Raising Vitamin–K–Dependent Coagulation Factors in Rabbits
Authors: Karpatkin Margaret , Karpatkin Simon
Published: March 2008

Summary. Evidence is presented for the existence of a humoral factor, obtained from rabbits treated with coumadin, which is capable of raising the vitamin‐K‐dependent coagulation factors II, VII, IX and X when injected into recipient animals. Thirteen rabbits were injected with varying volumes of test donor plasma (obtained from coumadin‐treated rabbits) over varying periods of time. A positive response in coagulation factor level (greater than 2 S.D. on 2 consecutive days) was obtained in 12 out of 13 animals for factor II, six out of eight animals for factor VII, six out of nine animals for factor IX and nine out of 13 animals for factor X. Ten rabbits were injected with control donor plasma from control rabbits injected with physiologic saline instead of coumadin. No risc was obtained in eight out of 10 rabbits for factor II nor in any of the animals examined for factors VII, IX and X (two, eight and seven animals respectively). The non‐vitamin‐K‐dependent coagulation factor V rose non‐specifically in four out of 11 recipient rabbits injected with test donor plasma and three out of nine injected with control donor plasma.
The dU Suppression Test using 125I‐UdR to Define Biochemical Megaloblastosis
Authors: Herbert Victor , Tisman Glenn , Go Le Teng , Brenner Lois
Published: March 2008

Summary. The deoxyuridine (dU) suppression test measures the de novo pathway to synthesis of DNA‐thymine. It demonstrates that in normoblastic human bone marrow the incorporation of triated thymidine (3H‐TdR) into DNA‐thymine is suppressed to less than 10% of control value by preincubation of bone marrow cells for 1 hr with deoxyuridine (dU), but in megaloblastic marrow cells there is less suppression.

In the current study it was demonstrated that 125I‐deoxyuridine (125I‐UdR) incorporation into DNA‐125I‐UdR may substitute for 3H‐TdR incorporation into DNA‐thymine in this test, thus simplifying the procedure by replacing a betaemitter with a gamma‐emitter.

Biochemical megaloblastosis was demonstrated or ruled out after incubation of human bone marrow for 3 hr with either tracer. The biochemical lesion present in folate or B12 deficiency can thus be recognized even in the absence of morphological change. The technique resolved questions raised by unclear bone marrow morphology and misleading serum vitamin B12 or folate levels, and is now routinely used in our laboratory on all bone marrow aspirates.
The Use of Platelet Serotonin Release as a Sensitive Method for Detecting Anti‐Platelet Antibodies and a Plasma Anti‐Platelet Factor in Patients with Idiopathic Thrombocytopenic Purpura*
Authors: Hirschman Richard J. , Shulman N. Raphael
Published: March 2008

Summary. The method described in this paper can be used for detecting anti‐platelet iso‐, hetero‐, and drug‐antibodies and for detecting a factor in the plasma of patients with idiopathic thrombocytopenic purpura that attaches to platelets (ITP factor). Plasma from the test subject, anticoagulated with citrate, is added to a suspension of normal platelets labelled with [14C] serotonin and the amount of radioactivity (serotonin) released is measured after incubating the mixture for 45 min at 37°C. Anti‐platelet antibodies and drug antibodies that fix complement as well as isoantibodies not detectable in vitro by complement fixation or agglutination can be measured by this test. Plasma from 24 of 40 patients with ITP released significantly more serotonin than control plasmas (P < 0.025). False positive results were obtained in less than 2.5% of plasmas from normal individuals, from patients with a variety of diseases and from patients with secondary thrombocytopenia. However, plasmas from two of 13 patients with systemic lupus erythematosus and normal platelet levels gave positive results in the test. The 7S gamma globulin fractions of plasmas containing isoantibodies or ITP factor as well as acid eluates from platelets which had been incubated in these plasmas gave positive serotonin‐release tests.
Methylmalonic Acid Excretion Studies
Authors: Chanarin I. , England J. M. , Mollin Caroline , Perry Janet
Published: March 2008

Summary. Oral valine and isoleucine were equally effective in producing an increased urinary excretion of methylmalonic acid (MMA) in vitamin B12 deficiency but oral MMA itself, methionine and leucine were unsatisfactory.

MMA excretion was increased in 16 out of 23 patients with vitamin B12 deficiency after valine and/or isoleucine loading but remained normal in seven patients. These latter patients tended to have the highest serum vitamin B12 levels and were the least anaemic. Thus, normal MMA excretion cannot be used as evidence to exclude vitamin B12 deficiency. The excretion was within normal limits in patients with hepatic cirrhosis and in patients with folate deficiency.
Detection of Genetic Variants of Haemophilia B with an Immunosorbent Technique
Authors: Neal W. R. , Tayloe D. T. , Cederbaum A. I. , Roberts H. R.
Published: March 2008

Summary. With an immunosorbent technique, we have found multiple genetic variants of haemophilia B. In one variant there is no detectable material that cross‐reacts with a human anti‐factor‐IX antibody. This variant is defined as negative for cross‐reacting material (CRM−). There is also a CRM+ variant in which the factor‐IX deficient plasma has the capacity to neutralize as much antibody as does normal plasma. There is an intermediate variety that neutralizes the specific antibody to a degree which is proportional to the factor‐IX clotting activity. This last group has been designated as having reduced cross‐reacting material (CRMR). The CRMR group is probably heterogeneous, and it is likely that many genetic variants of haemophilia B exist.
Effect of ADP‐induced Shape Change on Incorporation of 32P into Platelet Phosphatidic Acid and Mono‐, Di‐ and Triphosphatidyl Inositol
Authors: Lloyd J. V. , Nishizawa E. E. , Mustard J. F.
Published: March 2008

Summary. Experiments were carried out to determine whether the ADP‐induced increase in 32P‐content of phosphatidic acid (PA), monophosphatidylinositol (MPI), diphosphatidylinositol (DPI) or triphosphatidylinositol (TPI) of washed 32PO4 labelled platelets is related to platelet aggregation or to the change in shape induced by ADP. Washed platelets were labelled with 32PO4, washed, and resuspended in medium containing unlabelled PO4. With rabbit platelets, either the omission of Ca++ from the suspending medium or the addition of ethyleneglycoltetraacetic acid (EGTA) in amounts sufficient to chelate Ca ++ in the suspending medium, prevented ADP‐induced platelet aggregation. Lack of free Ca++ in the suspending medium did not prevent the ADP‐induced shape change or the increased labelling of PA and DPI. In suspensions containing EGTA, the increase in 32P‐content of PA induced by varying amounts of ADP was compared to the extent of the shape change. ADP in a final concentration of 10−6m or greater, produced both the maximum increase in 32P‐content of PA and the maximum degree of shape change. Washed platelets from pigs or humans without fibrinogen in the suspending medium showed a shape change following the addition of ADP but no aggregation, whereas in suspensions containing fibrinogen, the platelets showed both a shape change and aggregation. In suspension with and without fibrinogen, the ADP induced increase in 32P‐content of PA in human platelets and of PA+ MPI and DPI in pig platelets was the same. Prostaglandin E1 (PGE1), 10−4m final concentration, added to suspensions of rabbit platelets prevented both the ADP‐induced shape change and the increase in 32P incorporation into PA and DPI. PGE1, 10−4m, also caused a decrease in labelling of TPI. PGE1, 10−7m final concentration, prevented platelet aggregation but did not prevent the ADP‐induced shape change or the increase in 32P‐content of PA. Dibutyrylcyclic AMP (DBcAMP) inhibited both the ADP‐induced shape change and the increase in 32P‐content of PA. With rabbit platelets, the time course of the ADP‐induced shape change was compared to the time course of the increase in 32P‐content of PA, MPI and DPI. The increase in labelling of PA was detected 2 s after the addition of ADP and was near maximum at 8 s, the time of maximum shape change. The increase in labelling of DPI was not significant until 30 s. The regaining of the disc shape was associated with an increase in labelling of MPI and a decrease in labelling of PA and DPI. The results show that the changes in platelet PA are related to the ADP‐induced shape change rather than to aggregation.
Production of the Generalized Shwartzman Reaction in Rabbits by Ancrod (‘Arvin’) Infusion and Endotoxin Injection
Authors: Müller‐Berghaus Gert , Hocke Michael
Published: March 2008

Summary. The effect of the coagulant fraction from Agkistrodon rhodostoma venom (Arvin = ancrod) on the generation of microthrombi was studied in rabbits. Ancrod infusion caused hypofibrinogenaemia and induced the formation of circulating fibrin monomer complexes, but microthrombi were not detected in the glomerular capillaries. If, however, endotoxin was injected into ancrod‐infused rabbits, 73% of the animals showed microclots in the glomerular capillaries typical of the generalized Shwartzman reaction. These microclots exhibited antigenic characteristics of fibrinogen derivatives. Injection of endotoxin only did not produce glomerular capillary thrombosis and the mean fibrinogen levels did not decrease significantly, although fibrin monomer complexes could be detected in all the rabbits. Generation of microthrombosis by ancrod infusion and endotoxin injection could not be prevented by treating the animals with heparin. Thus endotoxin mediated the generation of microclots in ancrod‐infused rabbits by a mechanism different from its procoagulant activity. Heparin treatment interfered neither with the decrease in fibrinogen levels caused by ancrod infusion nor with the fall in the platelet counts caused by endotoxin injection.
Fine Structure and Peroxidase Activity of Circulating Micromegakaryoblasts and Platelets in a Case of Acute Myelofibrosis
Authors: Gorius J. Breton , Daniel M. T. , Flandrin G. , Denoel G. Kinet
Published: March 2008

Summary. In a case of acute myelofibrosis with thrombocytosis, electron microscopical examination of the buffy coat revealed the existence of numerous mononuclear cells with a blastic appearance. They were considered blasts because nucleoli were prominent, and ribosomes, free or linked to short saccules of rough endoplasmic reticulum (RER), were numerous. The presence of peroxidase activity in the perinuclear space and RER of these cells was identical with the occurrence and location of this enzyme reaction product in normal bone marrow megakaryocytes. The results indicated that the atypical cells were circulating micromegakaryoblasts. Highly abnormal platelets were also present in the patient's peripheral blood. The peroxidase test provides a useful adjunct for distinguishing cells of the megakaryocyte line which in the past may have been mistaken for atypical lymphocytes or myeloblasts by light or electron microscopy.
A Comparison of the Properties of Chicken Serum with Other Vitamin B12 Binding Proteins Used in Radioisotope Dilution Methods for Measuring Serum Vitamin B12 Concentrations
Authors: Newmark P. A. , Green R. , Musso A. M. , Mollin D. L.
Published: March 2008

Summary. The effects of pH, ionic strength, and vitamin B12 concentration on the vitamin B12 ‐binding capacities of chicken serum, normal human serum, chronic myelogenous leukaemia serum, and hog intrinsic factor concentrates are compared. The influence of pH on B12‐binding capacity was shown to be greatest at low pH and that of ionic strength was most evident below 0.1 ionic strength, particularly at acidic pH. The effect of B12 concentration on B12‐binding capacity was shown to vary with different binders, the pH of binding, and the time of incubation.

It was possible to select conditions for carrying out the binding of B12 to chicken serum so that the amount of B12 bound was virtually unaffected by the kind of variations in pH, ionic strength, or concentration of B12 that could occur in a radioisotope dilution assay of serum vitamin B12. Under these conditions it was further demonstrated that the binding was unaffected by the presence of denatured human serum. In addition chicken serum had a very great binding capacity and affinity for B12, was very stable on storage and was compatible with the use of protein‐coated charcoal.

Some conclusions on the optimal use of B12‐binding protein in the radioisotope dilution assay of serum vitamin B12 are made and the high potentiality of chicken serum in such a system is suggested.
A Red Cell Membrane Protein Abnormality in Hereditary Elliptocytosis
Authors: Gomperts E. D. , Cayannis F. , Metz J. , Zail S. S.
Published: March 2008

Summary. Membrane protein was extracted with acetic acid from the red cells of patients with hereditary elliptocytosis and subjected to electrophoresis on urea‐starch gel. Four different families were studied. Three patients from three families showed elliptocytosis in the absence of significant haemolysis. The membrane protein in these patients failed to demonstrate any alteration in electrophoretic pattern from the normal. Cases of elliptocytosis associated with severe haemolysis demonstrated an abnormal membrane protein electrophoretic pattern. This pattern differed within the same family varying with the severity of haemolysis. Thus one sibling with severe haemolysis demonstrated a grossly abnormal pattern while a less severely affected member showed an electrophoretic pattern similar to that of hereditary spherocytic red cells.
Linolenoyl Sorbitol and the Fragility of Hereditary Spherocytes
Authors: Livne Avinoam , Aloni B. , Moses S. , Kuiper P. J. C.
Published: March 2008

Summary. Linolenoyl sorbitol decreased uniformly the osmotic fragility of normal human red cells. When added to red cells of patients with hereditary spherocytosis (HS), the synthetic lipid afforded the most fragile cells the greatest shift in osmotic stability. The greater the osmotic fragility of red cells from different patients, the greater the protective effect afforded by the added lipid. Linolenoyl sorbitol fully reversed the increased fragility of vinblastin‐treated red cells. The results indicate a causal relationship between the lipid composition and the increased fragility of red cells in hereditary spherocytosis.
Quality Control Trials in the National Reference Thromboplastin Scheme
Authors: Leck Ian , Thomson Jean M. , Poller L.
Published: March 2008

Summary. In British hospitals routine measurements of prothrombin times are made either with a standardized thromboplastin reagent (the Manchester Comparative Reagent–MCR) or with the hospitals' own reagents, which have first been calibrated in terms of the national reference thromboplastin (the British Comparative Thromboplastin‐BCT). The fundamental assumption in the National Reference Scheme is that hospitals will obtain standardized results with the uniform technique and the BCT. Four collaborative studies have been conducted to evaluate this assumption. Instead of assessing their own local reagents hospitals were asked to calibrate given suspensions of unknown thromboplastin in terms of the BCT. In the last trial they were also asked to test three lyophilized plasma preparations. Since aliquots of given thromboplastin suspensions and dried plasma samples were distributed, all hospitals should have obtained similar results. There was, however, a wide variation, both in the ratios with the unknown thromboplastins that were equated with particular ratios with the BCT, and in the readings for the plasma samples. This variation was partly due to a tendency for some hospitals to obtain consistently higher readings than others. The situation might be improved by extending technical training in the prothrombin time test. If the differences between hospitals could be shown to be stable over long periods, a new system of quality control, involving the routine use of lyophilized plasma in addition to the BCT, might also be justified.
Erythrokinetic Studies as a Guide to the Value of Splenectomy in Primary Myeloid Metaplasia
Authors: Milner Gillian R. , Geary C. G. , Wadsworth L. D. , Doss A.
Published: March 2008

Summary. Red cell survival, plasma volume and erythrokinetic studies were carried out in 13 patients with progressive myeloid metaplasia. The patterns obtained were extremely variable. The majority had a haemodilutional anaemia, due to expansion of the plasma volume, which showed a rough correlation with splenic size. Although most patients showed evidence of haemolysis, only two had a severe haemolytic anaemia. Eight cases were selected for splenectomy; the best results were obtained in those in whom medullary erythropoiesis was still demonstrable by ferrokinetic studies pre‐operatively. In another two cases the spleen appeared to be the principal organ of erythropoiesis. Patients who had a haemodilutional anaemia pre‐operatively showed a substantial contraction in the plasma volume after splenectomy and, although this may not occur immediately after operation, it appears to be of long duration. It is suggested that the possibility of splenectomy be considered at an earlier stage in the evolution of the disease than has previously been customary.
Plasma Lysozyme in Drug‐induced and Spontaneous Cyclic Neutropenia
Authors: Hansen Niels Ebbe , Andersen Vagn , Karle Hans
Published: March 2008

Summary. In order to evaluate how well the plasma lysozyme concentration reflects changes in the neutrophil status in a non‐steady state system, six patients, treated with bone‐marrow depressing agents, were followed with neutrophil counts and plasma lysozyme determinations. In all patients, plasma lysozyme rather accurately mirrored changes in the neutrophil counts. The minimum change in neutrophil counts detectable by sequential studies of plasma lysozyme concentration was about 1000 neutrophils per μl blood, corresponding to a change in the granulocyte turnover rate of 30 × 107 neutrophils per kg per day. No time lag was found between changes in the neutrophil counts and plasma lysozyme concentrations, which suggests that the life span of the neutrophilic granulocytes in the tissues is less than 24 hr. In the neutrophil nadir phases the plasma lysozyme concentration was consistently higher than expected from the neutrophil counts. This was probably to a large extent due to ‘background’ lysozyme from non‐neutrophilic sources, but might in addition suggest a reduced life span in the neutropenic phases.

Similar investigations were carried out in two patients with spontaneous cyclic neutropenia. Whereas in these patients little variation of plasma lysozyme with the neutrophil phases could be demonstrated, plasma lysozyme was constantly higher than the concentrations found in the patients with drug‐induced neutropenia at similar neutrophil levels. This suggests a reduced neutrophil life span in the patients with cyclic neutropenia, especially in the nadir phases. Kinetic studies of blood neutrophils with DF32P labelling in one of the patients with cyclic neutropenia showed a slightly reduced blood half‐disappearance time in the zenith phase and a greatly reduced half‐disappearance time in the nadir phase.
Haemoglobin Perth: β32 (B14) Leu→Pro, An Unstable Haemoglobin Causing Haemolysis
Authors: Jackson J. M. , Yates A. , Huehns E. R.
Published: March 2008

Summary. An unstable haemoglobin, Hb‐Perth, β32 (B14) Leu→Pro, is described. The patient had a haemolytic anaemia with a T½51Cr RBC survival of 8 days, Hb 10–12 g/100 ml and had post splenectomy Heinz body formation.
The Effect of Ascorbate on the Maintenance of 2,3‐Diphosphoglycerate (2,3‐DPG) in Stored Red Cells
Authors: Wood L. A. , Beutler E.
Published: March 2008

Summary. The addition of adenine and ascorbate to citrate‐phosphate‐dextrose solution has been shown to prolong the maintenance of 2,3‐DPG levels of stored blood. The final concentrations of adenine and sodium ascorbate in the blood‐preservative mixture were 0.5 and 5.05 mm, respectively. In most cases, red cells stored in this preservative maintained levels of 2,3‐DPG greater than 50% of fresh values for 28 days. Red‐cell viability was not adversely affected by ascorbate. The mechanism by which ascorbate produced this effect is unknown.
Effects of Inflammation on Iron and Transferrin Metabolism
Authors: O'Shea M. J. , Kershenobich D. , Tavill A. S.
Published: March 2008

Summary. Experimentally induced acute inflammation in rats is associated with a fall in plasma iron and iron‐binding capacity. Transferrin synthesis by the liver of these animals is normal and not increased in the presence of a low serum iron, as it is in iron deficiency.

Human splenic macrophages and rabbit pulmonary macrophages which were maintained in culture for up to 1 week showed an ability to take up protein‐bound iron and labelled plasma proteins into the cytoplasm.

It is suggested that the macrophage may be the site both of iron sequestration and of transferrin degradation. This would provide a unified hypothesis to account for the finding of a low serum iron and TIBC in inflammatory disorders.
Assessment of the Value of Lysozyme Assay in Neutropenia
Authors: Levi J. A. , Macqueen Ann , Vincent P. C.
Published: March 2008

Summary. Estimations of the level of serum lysozyme were performed in 55 patients with neutropenia of various causes and bone‐marrow plasma lysozyme was measured in 39 of these patients. Neutrophil kinetic parameters (employing DF32P by an in‐vitro method) and serum lysozyme levels were measured at the same time in 30 of the neutropenic patients and 35 other subjects (14 healthy volunteers and 21 patients with non‐neutropenic disorders).

Levels of serum lysozyme were generally low when the bone marrow was hypo‐cellular and splenomegaly absent, and normal or high in the presence of a hyper‐cellular bone marrow and/or splenomegaly. However, no correlations were found between serum lysozyme and any of the neutrophil kinetic parameters measured, for the individual groups of the total group of subjects combined.

It appears that levels of serum and bone‐marrow plasma lysozyme depend on a number of different factors. Lysozyme levels may be of some help in the investigation of patients with neutropenia but cannot generally be used to derive any quantitative conclusions regarding the kinetic mechanisms involved in the production of neutropenia.