Quantitative behavior of eosinophil granulocytes in the bone marrow of children with acute lymphoblastic leukemia]

Of 31 children affected with acute lymphoblastic leukaemia the quantitative behaviour of eosinophilie granulocytes was examined in the course of the disease. Nearly all patients were treated according to a chemotherapy scheme (Memphis IV). During this therapy the eosinophils greatly diminished initially increased significantly to subnormal values and to the values of healthy persons with persisting full remission. Another significant decrease occurred during the relapse and in the pre-final stage. During each following relapse a greater diminution of bone-marrow eosinophils could be observed. Simultaneously the decrease of eosinophils led to a shift in the degree of maturation. In this connection the similar behaviour of neutrophilic and eosinophilic granulocytes of the bone-marrow must be stressed. Eventually, the lbast excrescence in the bone-marrow and its therapy cannot solely be decisive for the findings made. Relations to the lymphocytic system can be referred to.     

The ultrastructure of megathrombocytes in May-Hegglin's anomaly

A clinical and extensive electron microscopic study was made of the circulating granulocytes and thrombocytes from three female patients of a family with May-Hegglin anomaly. There have been observed inclusion bodies (D?hle-bodies) in a high proportion of polymorphonuclear leukocytes as well as a thrombocytopenia and giant platelets (so-called megathrombocytes). Other family members did not show hematologic abnormalities. The investigated giant platelets show ultrastructurally a different electron density of the alpha-granules, however, these structures present no typical formation of "bull's eye granules" which has been observed in other cases of May-Hegglin anomaly. Peripheral or central situated net-like structures of the endoplasmic reticulum have been found frequently. There were seen no microtubuli whereas glycogen particles were scattered within the peripheral cytoplasm.     

Hematologic complications in acute alcoholism]

The authors investigated the relations between cell anomalies (vacuolation, increasing sideroblastosis) caused by the uptake of alcohol and the dynamics of haematopoesis in the bone-marrow of 33 alcoholists who had been admitted in a comatose condition and who were neither affected with anaemia nor with chronic hepatitis. In all cases a maturation arrest of erythropoetic and granulopoetic cell elements which was not in accordance with the number of immature vacuolated cells could be observed. 12 test persons showed increased sideroblastic indices and a slightly diminished medullary reticulocytosis. The majority showed a very active thrombocytopoesis contrasting with the normal or even diminished number of thrombocytes in the peripheral blood. The authors come to the conclusion that alcohol will cause a general metabolic damage of haematopoesis and at the same time it will produce a direct toxic effect on the bone-marrow cells (proerythroblasts, promyelocytes) and the peripheral blood (thrombocytes).     

Experimental study of the organotropic properties of dactinomycin

Dactinomycin was studied pharmacologically on experimental animals. When dactinomycin was administered to the test-animals in doses close to the therapeutic ones for humans, suppression of the bone marrow blood formation was registered in spite of some increase in the number of the reticulocytes and thrombocytes in the peripheral blood and acceleration of the process of blood coagulation. In addition, the urea nitrogen blood levels increased. When the drug was administered in higher doses, suppression of the bone marrow blood formation was pronounced and the number of the leucocytes, reticulocytes and thrombocytes in the peripheral blood decreased. The rate of the blood coagulation decreased, while the biochemical values of the blood were indicative of impairement of the liver and kidney functions.     

The Breddin test in various forms of idiopathic thrombocytopenic purpura]

In 100 healthy persons and 38 patients with idiopathic thrombocytopenic purpura (ITP) the morphology of thrombocytes was examined according to BREDDIN. The survival time of the marked thrombocytes and their place of sequestration was determined in the patients and in 17 voluntary persons. A correlation was found to exist between the morphology of thrombocytes and those four forms of ITP which could be limited by means of radiometric methods. Moreover, a certain degree of dependence was found between the thrombocyte sequestration place and the presence of antibodies.     

Antibody-dependent cellular cytotoxicity (AAZZ) against leukemia cells--K-cell activity of human leukocytes

The activity of human leukocytes from the peripheral blood (PB) and bone-marrow (BM) to take the function as K-cells in the antibody dependent cellular cytotoxicity (ADCC) was tested in a 51Cr-test against mouse leukaemic cells and ALL cells covered with specific heterologous antibodies. Mononuclear PB-leukocytes and granulocytes of healthy donors and patients with leukemia and lymphoma in remission lysed murine and human leukaemic cells in the presence of specific antibodies. There was no lowering of K-cell activity of mononuclear PB-leukocytes of patients with leukaemia and lymphoma in remission under chemotherapy as compared with healthy donors and patients in remission without chemotherapy. There was a good correlation between the percentage of K-cell active mononuclear leukocytes in PB and BM. Attempts of fractionation with peripheral blood leukocytes of healthy donors resulted in the non-adherent mononuclear PB-leukocytes (lymphocytes) and granulocytes being effector cells in ADCC. To a high degree K-cell active lymphocytes could be identified in the non-B-fraction and only slightly in the fraction forming E rosettes.     

The state of hemopoiesis under conditions of long-term bone marrow irradiation in residents of the Techa riverside villages

The paper presents the results of a 50-year study on the state of hemopoiesis in the Techa riverside residents chronically exposed to radiation in the range from low to intermediate doses. The highest bone marrow doses were attributable to intakes of osteotropic 9OSr with drinking water and local food products. During the period of maximum radiation exposures (1951-1953) exposed residents were manifesting decreased counts of peripheral blood leukocytes (neutrophils and lymphocytes) and thrombocytes. Normal counts of erythrocytes were maintained owing to the effect of sufficient compensatory mechanisms, including accelerated rates of erythrocaryocyte proliferation and maturation. The development of peripheral blood granulocytopenia was influenced by the delay in the differentiation of neutrophilic bone-marrow granulocytes at the myelocyte phase, a marked increase in the frequency of lethal abnormalities in bone-marrow neutrophils, and pathological mitoses. The period of normalization of the blood cell composition was significantly variable for different blood cell series, and was noted to depend on exposure dose rate, extent of the primary hemopoiesis inhibition and specific physiological characteristics of exposed individuals.     

The incorporation of selenium and alterations of macro- and trace element levels in individual blood cells following supplementation with sodium selenite and vitamin E

Significant alterations in the selenium content of erythrocytes, thrombocytes, and neutrophil granulocytes were observed following a daily supplementation of 200 μg Se + 100 mg vitamin E during a period of 2 months. The neutrophil granulocytes incorporated more selenium than the thrombocytes. The iron content of the thrombocytes decreased on selenium supplementation, while the opposite was noted for the neutrophil granulocytes. The glutathione peroxidase activity was not significantly changed during the period of observation.     

Morphological characterization of the myelopoietic stem cell reserves in the human

According to microkinomatographic observations made on single cells up to ten days in vitro, there are the following laws of growth in haematopoiesis: Small cells will increase in growth up to five times in size, with their morphologic and kinetic properties being preserved, the blood lymphocyte will grow to immunoblasts, the small pluripotent stem cell to bone-marrow histiocytes. When growing, the myelopoietic stem cell may be gradually deviated into all myelopoietic cell lines. Instead of bone-marrow histiocytes it may differentiate to promyelocytes, promonocytes or proerythroblasts, all having an equal nucleus size, when it is induced by serum factors. Apart from histiocytes these large cells are capable of differentiating to a clon of blood cells, such as granulocytes, monocytes or reticulocytes, by several successive divisions of maturity. Contrary to the stimulated lymphocyte, symmetric mitoses will frequently occur, when small pluripotent stem cells are growing to bone-marrow histiocytes to be no further differentiated. Occasionally, asymmetric divisions may also be observed, i.e. one of the daughter cells will differentiate into one of the myelopoietic lines, whereas the other one will remain a progenitor cell. Moreover, there are various pathological mitoses in all progenitor cell sizes, such as endomitosis, cytoplasm fusion after mitosis, nucleus fusion after cytoplasm conjunction or amitotic nucleus division without cytokinesis. They produce megacaryoblasts further differentiating to megacaryocytes by corresponding pathological mitoses. According to our vital observations the pluripotency of the haematopoetic stem cell is being lost step by step.     

The function of granulocytes stored in whole blood in relation to the storage time

Whereas numerous investigations and extensive experiences have been available for some years on problems of storing erythrocytes and thrombocytes, it was only recently that the storage of granulocytes with the aim at maintaining their function for a longer period has been given proper regard as a problem, which, however, could not be brought to a satisfactory solution. Primarily, attention is focused on specific cell physiology. At the same time it is necessary to have at hand a number of approaches covering as many partial aspects of the granulocyte function as possible to identify the functional damages or the extent of cell damage in connection with storage. In the present paper the behaviour of granulocytes under conditions commonly used in transfusion medicine for storing blood are investigated for a period of 4 days by controlling the total leukocyte and granulocyte values, pH value, osmolality, vitality test in the colour excluding test as well as by function tests such as behaviour of phagocytosis and NBT capability of reduction. A report is presented on the results of quantitative and qualitative changes in the investigated parameters.     

Trout thrombocytes contain 12- but not 5-lipoxygenase activity

Fish thrombocytes are thought to be the evolutionary forerunners of mammalian platelets. Thrombocyte preparations made by conventional methods, such as density gradient centrifugation, contain other cell types such as neutrophilic granulocytes and lymphocytes that could interfere with subsequent experiments. In this study, rainbow trout thrombocytes were separated by density gradient centrifugation and further purified by magnetic cell sorting (MACS) using the thrombocyte specific monoclonal antibody, 30D8. Thrombocyte purity was assessed by reactivity to 30D8 using flow cytometry and immunocytochemistry. Following purification by density gradient centrifugation, thrombocytes were 66.9±9.2% (mean value±S.E.M., n=3) pure. Further purification by MACS significantly increased thrombocyte purity to 97.3±0.6%, whereas only 1.4% of the MACS ?ve fraction were identified as these cells. Incubation of thrombocytes isolated by density gradient alone with calcium ionophore, A23187, generated a range of eicosanoids derived from arachidonic or eicosapentaenoic acids, namely, leukotriene (LT)B₄, LTB₅, lipoxin (LX)A₄, LXA₅, 12-hydroxyeicosatetraenoic acid (12-HETE) and 12-hydroxyeicosapentaenoic acid (12-HEPE). A similar eicosanoid generation profile was observed for cells in the MACS ?ve fraction; however, MACS +ve cells (thrombocytes) generated no 4 or 5 series LT or LX but did generate significant amounts of the 12-lipoxygenase (LO) products, 12-HETE and 12-HEPE. These results indicate that trout thrombocytes contain no demonstrable 5-LO activity and like their mammalian counterparts possess 12-LO activity.     

Haematological and haematopoietic responses to starvation in an air-breathing fish Channapunctatus Bloch

Changes in haematological values (RBC numbers, haemoglobin content, haematocrit value, MCV, MCH, MCHC, TLC and DLC) based on weekly samples from a group of starved fish were investigated. After 8 weeks of starvation, the effects of restoration to a normal diet was evaluated. Parallel studies on haematopoietic tissues were also made. Changes in some biochemical values such as blood glucose, liver and muscle glycogen were also examined to correlate biochemical effects with those of haematological changes. Erythrocytes, thrombocytes and neutrophils were found to be most sensitive to starvation. The initial response to deprivation of food was an increase in RBCs and related values and in total leukocyte population. However, from week 5 onwards a sharp decline in these cell populations was noted. The leukocytes and thrombocytes showed a change parallel to RBC and the total leukocyte counts. However, neutrophils were observed to show a consistent increase throughout the starvation period. A blood glucose level below SOmglOOmh¹ appeared critical in relation to blood cell population. Haematopoietic studies revealed that reticulocytes and mesomyelocytes were unable to keep pace with the changing peripheral blood picture. Other stages in development responded to the changes in the peripheral blood.     

Prognosis in acquired aplastic anemia. An approach in the selection of patients for allogeneic bone marrow transplantation.

In a retrospective study, factors influencing the eventual course and outcome of patients suffering from aplastic anaemia were analysed. Of the 62 patients with pancytopenia concomitant with an aplastic or hypoplastic bone marrow, 36 have died during the first 24 months following diagnosis, 27 of these 36 cases within the first six months. 26 patients survived more than two years after presentation, but a further six of these cases succumbed during the following months. Of the 62 cases, 46 had neutrophil counts under 1 X 10(9)/l and platelet counts under 20 X 10(9)/l. These cases were subdivided according to their reticulocyte levels. In the age group of patients between 10 and 35 years, all those patients having more than 10,000/mul reticulocytes survived more than two years with only one exception, whereas only one of the 11 patients having reticulocyte counts below 10,000/mul has survived. In our series, the initial neutrophil and platelet counts were not of predictive value in identifying poor prognosis patients. The initial reticulocyte counts, however, appear to be a valid parameter for selecting patients in two groups, one with a favorable, and the other with an extremely unfavorable prognosis on conservative treatment. Young individuals with a poor predicted prognosis could thus be defined as possible candidates for bone marrow transplantation.     

Haematology and leucocyte morphology of wild caught Thunnus maccoyii

The haematology of wild southern bluefin tuna Thunnus maccoyii was described using blood samples collected from fish immediately after they were caught. Cytology and cytochemistry revealed that the blood in peripheral circulation is comprised of erythrocytes, reticulocytes, ghost cells, lymphocytes, thrombocytes, eosinophilic granulocytes, neutrophilic granulocytes and monocytes. Reference ranges established were 41·09–55·50% for haematocrit, 0·62–3·00% for leucocrit, 13·25–17·92 g dl⁻¹ for haemoglobin and 2·1–2·9 million erythrocytes μl⁻¹ for erythrocyte count. Differential cell counts showed 94·58 ± 2·15% erythrocytes, 3·99 ± 1·44% leucocytes and 1·43 ± 1·03% thrombocytes (mean ±  s . d .). Normal ranges for differential leucocyte counts were 0·00–5·45% for neutrophils, 0·69–12·06% for eosinophils, 0·00–5·03% for monocytes, 46·97–74·32% for lymphocytes and 14·47–43·92% for thrombocytes. Erythrocyte indices, leucocyte types and cytochemistry were comparable to other species of scombrids. Packed cell volume was sensitive to the physiological state of the fish and to sample handling technique.     

Identification of peripheral blood leucocytes of the dogfish (Scyliorhinus canicula L.) by electron microscopy

Dogfish peripheral blood leucocytes were examined by electron microscopy after the injection of colloidal carbon. The cells were classified as lymphocytes, plasma cells, monocytes, thrombocytes and granulocytes. The granulocytes were further classified into four types according to the structure of their granules. Monocytes, thrombocytes and two types of the granulocytic cells were phagocytic.     

Modification of uremic thrombocytopathy by hemodialysis

By means of in vitro screen pressure measurement the functions of thrombocytes were detected in dialysis patients before and immediately after dialysis. Start values are below the normal region as a sign of uraemic thrombocytopathia. The expected improvement after dialysis was not to be seen. As a cause of deterioration an activation of the thrombocytes at the surfaces of the capillary kidney and an incomplete inhibition of this process by heparin is discussed.     

Detection of alcohol abuse and monitoring of alcoholic abstinence using sideroblastic index and gamma-glutamyltransferase

45 hospitalised patients with chronic alcohol abuse observed immediately until that time preceding hospitalisation were examined with the aim of finding out whether examinations of the bone-marrow and gamma-glutamyltransferase (GGT) i.s. may be used for recording the consumption of alcohol and for monitoring the abstinence from alcohol. Bone-marrow puncture was made within 3 days after hospitalisation and was repeated at n = 35 after two weeks on an average. Simultaneously, GGT was determined. Disturbances of iron utilization, which were divided according to frequency and kind of sideroblasts into 4 degrees of seriousness, represented by far the most constant hematological findings. An sideroblastic+ index (SI) was counted, which, in addition to the count of sideroblasts, takes into account even qualitative disturbances. The sideroblastic+ index was increased in 91% (41/45) of patients irrespective of the presence or extent of an anemia so far as iron stores had not been completely depleted because of bleedings. In 71% (32/45) of the patients, gamma-glutamyltransferase (GGT) remained within the pathological range, thus lying significantly (p less than 0.05) below the sensitivity of the sideroblastic index (SI). By taking the increase of SI or GGT as a basis, the rate of recording alcoholics could be improved to 98% (44/45). Abstaining from alcohol caused a highly significant decrease of SI and GGT (p less than 0.005). Thereby, the sideroblasts index predominantly normalised in the period of examination, whereas gamma-glutamyltransferase fell below the pathological range only by way of exception. No significant decrease in the control value of SI and GGT was observed in those patients who did not abstain from alcohol. In comparing the differences of average values between abstaining and non-abstaining persons only SI revealed significant differences (p less than 0.005). SI and GGT complement each other in the control function of drinking behaviour. Under the given circumstances a simultaneous examination enables alcohol abuse to be recorded with nearly 100% of probability. SI is more sensitive and is able to differentiate more clearly between abstaining and non-abstaining. Due to its slower response GGT can indicate former alcohol abuse over a longer period. Concerning doubtful or potentially hepatotoxic+ substances at places of work, the sideroblastic+ index could provide an essential aid in deciding whether alcohol is a disturbing factor.     

Separation of leucocytes in the dogfish (Scyliorhinus canicula) using density gradient centrifugation and differential adhesion to glass coverslips

Summary The blood of the dogfish, S. canicula, contains several types of leucocytes, namely thrombocytes, monocytes, lymphocytes and four populations of granulocytes. Three of these granulocyte types, G1, G3 and G4, are eosinophilic while G2 is heterophilic/neutrophilic. All of the leucocyte types, with the exception of G2 granulocytes and monocytes, can be separated by means of their differential adherent properties to glass and by density gradient centrifugation. Thrombocytes, G3 and G4 granulocytes can be separated in good purity by single-step methods while G1 granulocytes and lymphocytes require a combination of density gradient centrifugation followed by adherence to glass to remove contaminating thrombocytes. Depending on the cell type, between 11–45% of cells with consistently high viability can be recovered after separation. Separated populations of the thrombocytes and granulocytes will be especially useful for studies on the role of such cell types in inflammation.     

Does ceftazidime or latamoxef affect the phagocytic capabilities of human granulocytes?

In comparative studies made in patients with pneumonia we have treated 14 cases with Latamoxef and 16 cases with Ceftazidime. We have focused attention on the side effects of these drugs and on the phagocytic capacity of granulocytes. In our patients we have noted a decrease of bactericidal activity of plasma and granulocytes and an elevated NBT reduction capability of resting granulocytes. The other tested function of the granulocytes exhibited only small variations. The applied therapy caused the bactericidal activity to be increased in granulocytes and plasma. Both drugs had no harmful effect on the phagocytic activities of granulocytes.     

Mitotic index in the bone marrow of children during the clinical course of acute lymphoblastic leukemia (ALL)]

From 31 children with acute lymphoblastic leukaemia the mitosis index in the bone-marrow was determined before the onset of therapy and during the clinical progress. Initially, the mean white mitosis index lay with 3.4% below that of the normal test persons, it rose significantly in the hematologic full remission and showed a decreasing tendency with a great range of dispersion in the recidive. The most lowered mitosis index was found in the final stage. Strong shifts in the kariologic distributions make a remaining in the prophase of the mitosis evident. The influence of polychemotherapy on the mitosis index and the phases of mitosis is discussed. Correlations between the mitosis index and clinical as well as paraclinical parameters were only to be found with respect to granulocytes and lymphoblasts. The considerable ranges of fluctuations of the mitosis index and the lack of congruity with the clinical progress of ALL allow no ensured assertions to be made for the single patient. It seems to be important in eosinophilia and in leukaemoid reactions.