Autoantibodies to the thrombocytes and erythrocytes in HIV-infected patients

In 48 patients with HIV infection were tested for the presence of autologous and allogenic antibodies to red blood cells with the use of Coombs' direct and indirect tests. 18 HIV-infected patients had IgG antibodies to thrombocytes, circulating in the blood (detected by the method of EIA) and bound to thrombocytes (detected by the method of RIA). In 5 out of 48 patients Coombs' direct test yielded positive results with red blood cells. 6 out of 18 examined patients had an elevated content of thrombocyte-bound antibodies. The presence of cross reactions between gp120 of human immunodeficiency virus and gp3a thrombocytes led to the formation of antithrombocyte antibodies and, consequently, to a decrease in the number of thrombocytes.     

Formation of Embryo Thromboblasts in Chick Blastoderm: Morphology, Site of Production and Time of Emergence in the Blood

Thrombocytes in the blood of chick embryos (termed embryo thrombocytes by L ucas and J amroz ) have PAS-positive granules in their cytoplasm. Electron microscopic observations reveal that the embryo thrombocytes contain glycogen granules present singly or in clumps. The presence of these inclusions and other morphological characteristics were used as specific markers to distinguish embryo thrombocytes from primitive erythroid cells. These markers also made it possible to determine the time at which the immature thromboblasts first emerge in blood vessels, and the period of their continued presence in the circulation. In this way we found that thromboblasts were detectable in embryos as early as stage 10⁺ of H amburger and H amilton (after 35 hr incubation) and that the thromboblasts were present in the circulation until day 4 of incubation (stage 23). In ovo and in vitro culture of de-embryonated blastoderm demonstrated that thromboblasts were formed in the area opeca vasculosa. The present observations suggest that embryo thromboblasts are formed at the same time and in the same area as the primitive cells of erythroid line.     

CHANGES IN SUSCEPTIBILITY OF CHICK EMBRYONIC BLOOD CELLS TO NEWCASTLE DISEASE VIRUS

To analyse the age-dependent changes in susceptibility of chick embryonic cells to viral infection, observations were made on the blood cells after the inoculation of Newcastle disease virus. A lethal dose of Sato strain of Newcastle disease virus was introduced into chick embryos via injection of inoculum into the blood vessel and allantoic sac. Observations of blood cells by immunofluorescent technique revealed two types of cells, susceptible and resistant to the virus infection. Erythroblasts of both primitive and definitive lines, embryonic thromboblasts and thrombocytes were of the former type and mid- and late-polychromatic erythrocytes and mature ones were of the latter. Erythroblasts gradually decrease in their viral susceptibility as they differentiate into polychromatic erythrocytes and finally become resistant to the virus at the mid-polychromatic erythrocyte stage or later. Thromboblasts, on the other hand, retain high susceptibility to the virus throughout the course of their differentiation to mature thrombocytes. The change in the viral susceptibility of erythroid cells with differentiation is discussed in relation to the structural and functional alterations during the cell specialization.     

Shape transformation and cytoskeletal reorganization in activated non-mammalian thrombocytes

The nucleated thrombocytes of non-mammalian vertebrates are partially flattened, ovoid cells morphologically distinct from mammalian platelets, and the extent of their functional equivalence is unknown. To test whether they resemble platelets in having similar F-actin-based post-activation stages, rapid fixation/extraction/labeling methods were developed to reveal cytoskeletal organization in dogfish thrombocytes by confocal microscopy. Unactivated cells contained cortical F-actin plus denser F-actin co-localizing with outer marginal band (MB) microtubules. In the post-activation sequence, determined for the first time by continuous observation of individual thrombocytes following thrombin perfusion, cells rounded and blebbed, spread, and eventually flattened extensively. The MB twisted and then became disorganized, with microtubule bundles remaining centrally located and associated with nuclear clefts. In contrast, F-actin occupied blebs and outward-spreading cytoplasm, initially in spiky projections, then predominantly in stress fibers, and inhibitors of F-actin assembly or myosin ATPase blocked shape changes. Thus, the post-activation stages and cytoskeletal events observed in nucleated thrombocytes were found to parallel those of platelets.     

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.     

Quantitative analysis of shape and volume changes in activated thrombocytes in real time by single‐shot spatial light modulator‐based differential interference contrast imaging

We suggest to use a combination of optical tweezers and single‐image quantitative differential interference contrast (DIC) emulated by a spatial light modulator (SLM) to study physiological shape changes in thrombocytes after activation and demonstrate the effectiveness of this system for the given task. A specially designed phase mask displayed at the SLM enables quantitative phase calculation from only a single recording. The optical tweezers stabilize trapped thrombocytes for long‐time monitoring of changes in the optical thickness profile of thrombocytes during activation by adenosine diphosphate (ADP). (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Results of a new platelet function test in various clinical diseases and in preserved blood]

In healthy persons, patients with thrombocytopenia and patients with arterial disturbances of blood flow with and without administration of Falithrom the pressure recording was carried out after the combined aggregation and adhesion of thrombocytes. The reaction time particularly observed after administration of ADP revealed marked differences in the groups examined, likewise the maximal pressure amplitude in front of the pumped through filter. When Falithrom was administered the thrombocyte functions were clearly impaired. The reduction of the capacity of platelets to obstruct filter pores can impressively be proved even in those thrombocytes stored in ACD-AG-plasma.     

Mechanisms of the haematological changes induced by hyperventilation

During voluntary hyperventilation an increase in the lymphocyte and thrombocyte counts occurs, paralleled by an increase in plasma epinephrine and norepinephrine. All these changes are rapidly reversible after hyperventilation and are followed by an increase in the neutrophil granulocyte count. The pathophysiological mechanisms of these changes were investigated by comparison of the hyperventilation-induced changes of the blood picture in 11 normal, 9 splenectomized and 12 beta-blocked volunteers. Splenectomy did not affect the hyperventilation-induced mobilization of lymphocytes and neutrophils but totally suppressed the change in the thrombocyte count. beta-blockade by 80 mg propranolol did not suppress the hyperventilation-induced increase in neutrophils. It reduced the absolute increase of lymphocytes and thrombocytes by half, but it also increased the baseline counts of these cells. The study shows that hyperventilation mobilizes thrombocytes from the spleen but not from extralienal pools, and that lymphocytes and neutrophils are mobilized from extralienal pools. Whereas neutrophil mobilization is not suppressed by beta-blockade, the reduction of hyperventilation-induced mobilization of lymphocytes and thrombocytes may be due to a reduction in the size of the mobilizable cell pools, and therefore cannot be interpreted as a sure indication that adrenergic mechanisms are involved in their hyperventilation-induced mobilization.     

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.     

Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals

The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not detectable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase.     

Ultrastructural distribution of peroxidase in thrombocytes of mammals and submammals

Summary The localization and distribution of peroxidase (PPO) activity were studied ultracytochemically in thrombocytes from lampreys, carps, frogs, snakes, tortoises, rabbits, sheep, dogs, and monkeys. PPO activity was not deteetable in the thrombocytes of lampreys, carps, frogs, and snakes. However, this enzyme activity was demonstrated in the nuclear envelope and endoplasmic reticulum of tortoise thrombocytes. Dog and monkey thrombocytes (blood platelets) exhibited PPO activity in the dense tubular system, but this enzyme activity was not detectable in rabbit and sheep thrombocytes. Our observations are interpreted to suggest that thrombocytes from animals lower than amphibia are peroxidase negative. Furthermore, it can be said that thrombocytes from animals higher than reptiles are generally positive, although there are exceptions. PPO activity was localized in the endoplasmic-reticulum system, but not in the cytoplasmic granules of thrombocytes common to submammals and mammals. In this study, we also compared the distribution of peroxidase activity in thrombocytes, neutrophils, and eosinophils and conclude that these are significant differences in the distribution of PPO and myeloperoxidase.     

Chicken thrombocytes in culture: lymphocyte-conditioned medium delays apoptosis.

Chicken thrombocytes are nucleated cells, analogs to mammalian platelets. These cells are involved in hemostasis, phagocytosis and secretion of specific products. Most of the properties of avian thrombocytes have been established in experiments that employed recently isolated blood cells. Attempts to cultivate these cells for a long period of time under optimal culture conditions for peripheral blood cells were unsuccessful; thrombocytes died after 24 h of cultivation unlike macrophages cocultured with them. Here we investigate the reasons and type of thrombocyte death in culture. Thrombocytes were separated from peripheral blood of roosters and cultured for 48 h. The influence of different culture conditions on thrombocyte viability was studied. Cells were cultured as adherent cell monolayers or under agitation (preventing adherence), in the presence or lack of lymphocytes or their soluble factors, and various concentrations of fetal bovine serum. After 24 h in standard culture thrombocytes displayed cytoplasm and chromatin condensation, DNA cleaved into oligonucleosomal fragments and unaltered mitochondria. These results strongly suggest that thrombocytes suffer an apoptotic cell death in culture. Apoptosis could be delayed by culturing thrombocytes in the presence of lymphocytes or their soluble factors.     

Composition of main haematopoietic compartments in normal and bled channel catfish

Differential cell counts showed that the head and trunk kidney of control and bled channel catfish Ictalurus punctatus had myeloid characteristics. They contained lymphoid and granuloid cells, thrombocytes, erythroid and agranular cells in decreasing order of abundance (%). Among the blast and precursor cells, the most numerous erythroid ones were followed by granuloid, lymphoid and agranular ones. The main changes after blood withdrawal were the decrease of thrombocytes and the increase of precursor cells in both kidney parts. In the group examined 7 days after bleeding the head kidney had a higher percentage of erythroid cells and lymphocyte precursors than the trunk kidney while the latter had more granuloid cells and their precursors. Basophils were present ( c . 1%) in both regions of the kidney of all groups. The spleen was predominantly a lymphatic organ. It contained c . 80% lymphoid cells, a higher incidence of granulated lymphocytes than in kidneys, 15% thrombocytes and 1.4% agranular cells. Blood withdrawal caused an increase of thrombocytes, a decrease of lymphoid cells and an increase of erythroid precursors in the spleen. The last probably stemmed from the circulation. While haematocrit values failed to indicate the anaemic state in the bled groups, the differential red blood cell count showed dramatic differences between the control and bled groups as well as between the two groups in different stages of recuperation from the blood loss.     

2种病原菌感染海鳗后外周血白细胞的病理变化

用鳗弧菌(Vibrio anguillarum)和嗜水气单胞菌(Aeromonas hydrophila)分别感染海鳗(Muraenesoxcinereus),48 h后取其外周血液,瑞-吉(Wright-Giemsa)氏染色,光镜下观察,同时对其外周血液中的免疫细胞数量变化进行测定。鳗弧菌和嗜水气单胞菌感染组的单核细胞的伪足样突起与对照组比较明显增多,胞质内空泡增多、变大,数量明显增多,差异极显著;嗜中性粒细胞核多呈长杆状,数量与对照组比较明显增多,差异极显著;淋巴细胞伪足样突起明显,中、大型淋巴细胞较多,数量与对照组相比明显减少,差异极显著;血栓细胞的数量明显下降,差异极显著。     

A new method for the determination of thrombocyte function in patients and in preserved blood]

In a circulating thrombocytic plasma the pressure in front of a defined mesh filter depends on the combined capacity of adhesion and aggregation of thrombocytes. A new clinical in vitro test is based on this principle which comes nearest to the conditions to be found in hemostasis dependent on thrombocytes. Judging from the preliminary results of investigations performed on stored blood, blood donors, patients with thrombocytopenia, disturbances of blood flow and patients under anticoagulant therapy, the time until the pressure rise after administering ADP and the maximal pressure amplitude proved to be the most significant parameters of the new procedure.     

Isoenzymes of acid phosphatase in blood cells of normal subjects and patients with leukemia (author's transl)]

The activities of acid phosphatases (AP) were measured in leukocytes from patients with chronic myelocytic leukemia (CML), macrophages, granulocytes, in the fractionated mononuclear cells of patients with CML and with hairy-cell-leukemia (HCL) and in the cells from patients with acute leukemia (AL). The lowest activities were found in lymphocytes of normal subjects and of patients with chronic lymphatic leukemia (CLL) and in thrombocytes. Isoenzyme (IsE) 1 was characteristic for thymocytes, IsE 2 for granulocytes, IsE 3 for pathologic blast-cells, lymphocytes and thrombocytes, IsE 4 for macrophages, IsE 5 with components a and b for the mononuclear fraction of patients with HCL. In addition IsE 5 was detected in lymphocytes, macrophages and CLL-cells. In 4 patients with HCL the relative percentage of IsE-5-fraction was slightly greater than the percentage of tartrate resistant cells. In two patients with questionable HCL well marked IsE-5-fractions were recognized but no tartrate resistant cells. In one patient with HCL a relatively high percentage of tartrate resistant hairy-cells and in comparison an inadaquate low IsE-5-fraction was found. These different relations were explained with the more sensitive method of gelelectrophoresis and different affinity of substrates to AP.     

Postoperative changes of the thrombocyte number and function]

In 30 patients the number of thrombocytes was determined 24-48 hours after surgical interventions and compared with the normal range. The function of thrombocytes was determined by the method of pressure registration in combined thrombocyte-aggregation-adhesion (DKTA method). In spite of the occurring thrombocytosis there is a tendency towards a decrease of response in most cases and thus of haemostatic function of blood platelets. The influence of the thrombocyte function caused by fibrinolytic split products or by a change of prostaglandine metabolism is discussed.     

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.     

Changes in various blood platelet function tests in rabbits exposed to whole-body irradiation

With a selected spectrum of coagulation tests the functioning capacity of thrombocytes was investigated in rabbits exposed to a whole body irradiation by means of 60Co radiation with a LD 5/30. A reduced retraction could be proved for times of irradiation (the 5th, 8th, 11th, 21st, 35th, and 56th day). A reduced formation of malondialdehyde could be identified in thrombocytes on the 8th and 21st day after irradiation. No changes could be found in determining adhesiveness, platelet aggregation caused by ADP, and PF3A and PF3F tests. In the course of additional investigations (coagulation time in unprepared and siliconized glass tubes, thromboelastogramme, activated partial chromboplastine time), significant changes of coagulation time could be observed in siliconized glass tubes on the 8th, 11th, 21st, and 56th day following irradiation.     

Regulation of thrombocytopoiesis studied by a mathematical model

A mathematical model of thrombocytopoiesis is proposed which accounts for the recent data on its regulation. It is shown that the compensatory response of the system to a decrease in the level of thrombocytes in the blood is controlled by the total amount of thrombocytes and megakaryocytes. The proliferation intensity of megakaryocytes and the total number of thrombocytes reveal, respectively, a lineary and a logarithmical dependence on the total number of thrombocytes and megakaryocytes. The limits of the post-transfusion level of thrombocytes are defined, within which the thrombocytopoiesis is controlled only by the number of thrombocytes. The values of parameters characterizing the behaviour of the thrombocytopoiesis system are calculated.