Hematological Parameters Influencing the Thrombostat 4000

1995 ◽  
Vol 21 (S 02) ◽  
pp. 20-24 ◽  
Author(s):  
Dietmar Söhngen ◽  
Eckhard Hatistein ◽  
Axel Heyll ◽  
Gerald Meckenstock ◽  
Simone Wienen ◽  
...  
Keyword(s):  
Linear Correlation ◽  
Polymorphonuclear Leukocytes ◽  
Bleeding Time ◽  
Hematological Parameters ◽  
Chronic Myelocytic Leukemia ◽  
Platelet Counts ◽  
Myelocytic Leukemia ◽  
Closure Time ◽  
Leukocyte Counts

The effects of platelet counts, hematocrit, and leukocyte counts were studied on the closure times of the Thrombostat 4000 (in-vitro bleeding time, IVBT). Closure times became longer with platelet counts <50 × 109/L; an inverse linear correlation could be established. Hematocrit was also inversely correlated with the closure time. At constant platelet counts a hematocrit of 55% yielded an immediate closure of the filter, while with a hematocrit <15% no closure times could be measured. At constant platelet counts and hematocrits, nomonuclear and polymorphonuclear leukocytes also influenced closure times; increased counts resulted in shorter closure times. Leukocytes from a patient with chronic myelocytic leukemia had the same effects.

scholarly journals Constitutive expression of GATA-1, EPOR, alpha-globin, and gamma-globin genes in myeloid clonogenic cells from juvenile chronic myelocytic leukemia

Blood ◽  
1995 ◽  
Vol 86 (1) ◽  
pp. 323-328 ◽  
Author(s):  
E Privitera ◽  
R Schiro ◽  
D Longoni ◽  
A Ronchi ◽  
A Rambaldi ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Leukemic Cells ◽  
Globin Genes ◽  
Chronic Myelocytic Leukemia ◽  
Beta Globin ◽  
Globin Mrna ◽  
Gamma Globin ◽  
Myelocytic Leukemia ◽  
Alpha Globin

Juvenile chronic myelocytic leukemia (JCML) is a rare disorder of early childhood. Characteristic of JCML are the progressive appearance of high levels of fetal hemoglobin (HbF), reflecting a true reversion to a fetal type of erythropoiesis, and the presence of colony-forming cells able to grow in vitro spontaneously in the absence of growth factors. To better understand the relationship between the erythroid abnormalities and the leukemic process, we analyzed the expression pattern of specific genes related to erythroid differentiation--GATA-1, EPOR, alpha-globin, beta-globin, and gamma-globin genes--in JCML peripheral blood (PB) cells and in vitro-derived colonies. Northern blot analysis of PB cells from five JCML patients indicated levels of GATA-1 transcripts much higher than those usually found in other types of leukemic cells, and S1 nuclease protection assay detected significantly increased expression of gamma-globin mRNA. Reverse transcription-polymerase chain reaction (RT-PCR) analysis of single granulocyte-macrophage colony-forming unit (CFU-GM) colonies, obtained in vitro in the absence of added growth factors from four JCML patients, detected GATA-1, EPOR, and globin (alpha and gamma) transcripts in most of the colonies tested, in contrast with control CFU-GM from normal bone marrow, which were positive only for GATA-1. Single JCML colonies were tested for the presence of two different transcripts; whereas alpha- and gamma-globin genes appeared mostly coexpressed, beta-globin mRNA was detected only in a minority of the gamma-globin-positive colonies, indicating that the leukemic pattern of hemoglobin synthesis is mainly fetal. In addition, the leukemic cells occurring during blast crisis of one of our patients displayed the typical features of a stem cell leukemia (CD34+, CD19-, CD2-, myeloperoxidase-). In this sorted CD34+ population, we detected the presence of a marker chromosome, der(12)t(3;12), previously identified in bone marrow cells at diagnosis and an expression pattern superimposable to that of the JCML colonies, consistently displaying a high gamma-globin:beta-globin mRNA ratio. The expression of erythroid markers within populations of leukemic cells, both in vivo and in vitro, supports the hypothesis that abnormal JCML erythroid cells may originate from the same mutated progenitor that sustains the growth of the leukemic cells.

scholarly journals Serial in vitro bone marrow fibroblast culture in human leukemia

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 589-592 ◽  
Author(s):  
T Nagao ◽  
K Yamauchi ◽  
M Komatsuda
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Colony Formation ◽  
Calf Serum ◽  
Culture Method ◽  
Fibroblast Culture ◽  
Human Leukemia ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia

Abstract Human fibroblast colony formation from bone marrow was performed in liquid culture. Fetal calf serum was used as a stimulator of the fibroblast colony formation. The colony formation took place not only in normal donors, but also in patients with acute leukemia and chronic myelocytic leukemia. At the diagnosis of the disease, significant colony suppression was observed in most cases of acute leukemia, while the number of colonies increased in half of the cases of chronic myelocytic leukemia. However, there was no correlation between the colony-forming efficiency and the initial number of peripheral platelets or bone marrow megakaryocytes that contained growth-promoting factor. The number of colonies increased after chemotherapy, recovered at the stage of complete remission, and then decreased to low levels at relapse in the patients with acute leukemia; it decreased after treatment with busulfan in the patients with chronic myelocytic leukemia. This fibroblast culture method is useful for counting fibroblast colony-forming cells in the bone marrow of human leukemia.

scholarly journals Chronic myelocytic leukemia (CML): failure to detect residual normal committed stem cells in vitro

Blood ◽  
1979 ◽  
Vol 53 (2) ◽  
pp. 264-268 ◽  
Author(s):  
JW Singer ◽  
PJ Fialkow ◽  
L Steinmann ◽  
V Najfeld ◽  
SJ Stein ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Type A ◽  
Normal Subjects ◽  
Myeloproliferative Disorder ◽  
Chronic Myelocytic Leukemia ◽  
Type B ◽  
Normal Tissues ◽  
Myelocytic Leukemia

Abstract Granulocytic colonies grown in culture from marrow and peripheral blood from five patients with Ph1-positive CML and heterozygous at the G-6-PD locus were analyzed for G-6-PD in order to identify CFU-C that do not arise from the CML clone. The patients had both B and A enzymes in normal tissues, but their CML clones typed as B. Whereas about 50% of colonies from normal subjects heterozygous as the G-6-PD locus show type-A G-6-PD and 50% type B, only two of the 1308 colonies from the CML patients had type-A G-6-PD. These data provide little evidence for persistence of normal committed stem cells in CML, a finding in contrast to that made previously in polycythemia vera, another clonal stem cell myeloproliferative disorder.

Functions of Glycosylation Cooled Rabbit Platelets In Vivo and In Vitro.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3902-3902
Author(s):  
Bao-An Chen ◽  
Cheng-Yin Huang ◽  
Xiao-Ping Pei ◽  
Chong Gao ◽  
Jia-Hua Ding ◽  
...  
Keyword(s):  
Survival Time ◽  
Bleeding Time ◽  
Rabbit Platelet ◽  
Control Group ◽  
Platelet Counts ◽  
Rabbit Ear ◽  
Significant Difference ◽  
Rabbit Platelets

Abstract This study was aimed to investigate functions of glycosylation cooled rabbit platelets in vivo and in vitro and the method to store cold platelets with UDP-gal. We collected rabbit heart blood, prepared concentrated platelet suspensions in a normal way to which we added UDP-Gal, and then stored them for ten days in 4° refrigerator. Thereafter platelet counts, mean platelet volume, platelet distributing width, platelet aggregation function, the activity to urge coagulation including PF3aT and APCT and apoptosis were determine- d. Meanwhile, survival time in vivo was tested after cold-stored rabbit platelets labeled with Cr51 were transfused into rabbits. Rabbit ear bleeding time and percentage plate recovery(PPR) were determined 1 hour and 24 hour after they were transfused into rabbit thrombocytopenia model. Results show that there was not significant difference in PLT counts, MPV, PDW, PF3aT and APCT between UDP-Gal cold-stored platelet group and fresh platelet group(p>0.05). On the contrary, platelet counts decreased significantly, MPV, PDW jumped and PF3aT and APCT went down in cold control group compared to fresh platelet group(p<0.01). Apoptosis increase in UDP-Gal cold-stored platelet group compared with fresh platelet group(p<0.05), but was significantly lower than that in cold control group(p<0.01). Although PagT(inducing reagent: C-PG) decreased, it could still be above 50% of fresh platelets. Survival time in rabbit vivo was close between UDP-Gal cold-stored platelet group and fresh platelet group(p<0.05). Survival rate seventy-two hours after transfusion in fresh platelet group, UDP-Gal cold-stored platelet group and cold control group was 57.5%±7.2%,50.3%±6.3% and 0.1%±0.1% respectively. Rabbit ear bleeding time was significantly shortened after transfusion of galactosylation cooled rabbit platelet (p<0.01), In contrast, it had less change in cold control group(p>0.05). PPR was 66.1%±0.5%,47.8%±0.6%;60.9%±0.3%,41.6%±0.4%;47.7%±0.5%,9.4%±0.5% respectively in fresh platelet group, UDP-Gal cold-stored platelet group and cold control group. PPR after transfusion of galactosylation cooled rabbit platelet had no statistical difference compared with that of fresh platelet group(p>0.05), and they in both groups were much higher than that in cold control group(p<0.01). Conclusion: Galactosylation can improve functions of cooled rabbit platelets in vivo and in vitro. and prolong the storage time of them.

THE BIOSYNTHESIS FROM ACETATE-1-C14 OF FATTY ACIDS AND CHOLESTEROL IN FORMED BLOOD ELEMENTS

1958 ◽  
Vol 36 (1) ◽  
pp. 1125-1136 ◽  
Author(s):  
V. J. O'Donnell ◽  
P. Ottolenghi ◽  
A. Malkin ◽  
O. F. Denstedt ◽  
R. D. H. Heard
Keyword(s):  
Fatty Acids ◽  
The Other ◽  
Chronic Myelocytic Leukemia ◽  
Monocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Lymphatic Leukemia ◽  
Thymus Tissue ◽  
Mammalian Blood ◽  
Blood Elements

The biogenesis, in vitro, of cholesterol and fatty acids from acetate-1-C14 in the cellular elements of blood has been studied. From the results obtained it is apparent that whereas the normocytes (nucleated) from the chicken and from the mammal (nonnucleated) are devoid of the capacity to synthesize cholesterol and fatty acids, the reticulocytes and the normoblasts from the rabbit, with an experimentally produced reticulocytosis, possess the ability to synthesize these lipids. Studies on the leucocytes from normal mammalian blood and thymus tissue showed little or no activity. On the other hand, the leucocytes from the thymus tissue and blood of chickens and from the blood of a patient with chronic myelocytic leukemia and from another with monocytic leukemia (Naegeli type) were capable of utilizing CI4-acetate for the synthesis of lipids. The leucocytes from patients with chronic lymphatic leukemia were found to be devoid of this activity.

scholarly journals Effect of Myleran and 6-Mercaptopurine (6-MP) on the Serum Level of Vitamin B12 in Chronic Myelocytic Leukemia

Blood ◽  
1959 ◽  
Vol 14 (3) ◽  
pp. 274-278 ◽  
Author(s):  
V. M. DOCTOR ◽  
D. E. BERGSAGEL ◽  
C. C. SHULLENBERGER
Keyword(s):  
Serum Level ◽  
Vitamin B12 ◽  
Serum Vitamin ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Leukocyte Counts

Abstract Serum vitamin B12 determinations and the total leukocyte counts were made at regular intervals in chronic myelocytic leukemia patients before, during and after therapy with 6-MP or Myleran. The results indicate that a reduction in total leukocytes following therapy is followed by a gradual lowering of serum vitamin B12. The levels of serum vitamin B12 at the completion of therapy may be related to the duration of remission.

scholarly journals Serial in vitro bone marrow fibroblast culture in human leukemia

Blood ◽  
1983 ◽  
Vol 61 (3) ◽  
pp. 589-592 ◽  
Author(s):  
T Nagao ◽  
K Yamauchi ◽  
M Komatsuda
Keyword(s):  
Bone Marrow ◽  
Acute Leukemia ◽  
Colony Formation ◽  
Calf Serum ◽  
Culture Method ◽  
Fibroblast Culture ◽  
Human Leukemia ◽  
Chronic Myelocytic Leukemia ◽  
Myelocytic Leukemia

Human fibroblast colony formation from bone marrow was performed in liquid culture. Fetal calf serum was used as a stimulator of the fibroblast colony formation. The colony formation took place not only in normal donors, but also in patients with acute leukemia and chronic myelocytic leukemia. At the diagnosis of the disease, significant colony suppression was observed in most cases of acute leukemia, while the number of colonies increased in half of the cases of chronic myelocytic leukemia. However, there was no correlation between the colony-forming efficiency and the initial number of peripheral platelets or bone marrow megakaryocytes that contained growth-promoting factor. The number of colonies increased after chemotherapy, recovered at the stage of complete remission, and then decreased to low levels at relapse in the patients with acute leukemia; it decreased after treatment with busulfan in the patients with chronic myelocytic leukemia. This fibroblast culture method is useful for counting fibroblast colony-forming cells in the bone marrow of human leukemia.

THE BIOSYNTHESIS FROM ACETATE-1-C14 OF FATTY ACIDS AND CHOLESTEROL IN FORMED BLOOD ELEMENTS

1958 ◽  
Vol 36 (11) ◽  
pp. 1125-1136 ◽  
Author(s):  
V. J. O'Donnell ◽  
P. Ottolenghi ◽  
A. Malkin ◽  
O. F. Denstedt ◽  
R. D. H. Heard
Keyword(s):  
Fatty Acids ◽  
The Other ◽  
Chronic Myelocytic Leukemia ◽  
Monocytic Leukemia ◽  
Myelocytic Leukemia ◽  
Lymphatic Leukemia ◽  
Thymus Tissue ◽  
Mammalian Blood ◽  
Blood Elements

The biogenesis, in vitro, of cholesterol and fatty acids from acetate-1-C14 in the cellular elements of blood has been studied. From the results obtained it is apparent that whereas the normocytes (nucleated) from the chicken and from the mammal (nonnucleated) are devoid of the capacity to synthesize cholesterol and fatty acids, the reticulocytes and the normoblasts from the rabbit, with an experimentally produced reticulocytosis, possess the ability to synthesize these lipids. Studies on the leucocytes from normal mammalian blood and thymus tissue showed little or no activity. On the other hand, the leucocytes from the thymus tissue and blood of chickens and from the blood of a patient with chronic myelocytic leukemia and from another with monocytic leukemia (Naegeli type) were capable of utilizing CI4-acetate for the synthesis of lipids. The leucocytes from patients with chronic lymphatic leukemia were found to be devoid of this activity.

Thrombocytopenia-Adapted In Vitro Bleeding Test Assesses Platelet Function in Thrombocytopenic Patients

1995 ◽  
Vol 21 (S 02) ◽  
pp. 79-90 ◽  
Author(s):  
Volker Kretschmer ◽  
Bernd Huss ◽  
Gerrit Bonacker ◽  
Jörg Hoffmann ◽  
Simke Bewarder ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Clinical Study ◽  
Platelet Function ◽  
Bleeding Time ◽  
The Other ◽  
Autoimmune Thrombocytopenia ◽  
Distribution Width ◽  
Platelet Counts ◽  
Cell Counts

Platelet counts do not always reflect the true hleeding risk in chronically thrombocytopenic patients, and the posttransfusion platelet increments do not necessarily demonstrate the therapeutic efficacy. There are no easy and reliable tests yet permitting the determination of platelet function in thrombocytopenic patients. The in-vitro bleeding test (IVBT) with the Thrombostat 4000 proved to be a very sensitive and specific test for the detection of platelet disorders. In order to become suitable for the investigation of thrombocytopenic blood with platelet count between 5 × 109/L and 50 × 109/L, special modifications were necessary. We report on the evaluation of two thrombocytopenia-adapted modifications (TP-IVBT 150/120), first with blood of healthy donors made thrombocytopenic (three experiments with six blood samples each of different platelet concentrations and identical hematocrit) and then in a clinical study on 77 thrombocytopenic patients (69 with bone marrow hypoplasia, eight with autoimmune thrombocytopenia) receiving 267 platelet transfusions. The patients were followed over 15 days on average (1–67 days) by daily examinations (total 1,285 observation days). Most TP-IVBT measurements were carried out in triplicate, using the modification with the 120 μm filter (TP-IVBT 120) because it proved to be superior to the other modification. Additionally, cell counts, hematocrit, body temperature, platelet volume, platelet distribution width, expression ofCD 36, 41a, 42b on platelets, Simplate® bleeding time, and detailed analysis of bleeding signs were performed for the calculation of a bleeding score. There was a close correlation between TP-IVBT and platelet counts with thrombocytopenic normal blood (r2 = 0.81 − 0.94). This indicated the suitability of this test modification to examine platelet function in thrombocytopenic patients. The clinical study showed that the TP-IVBT helped at least to determine the platelet-related bleeding risk in thrombocytopenic patients. It allowed differentiation between hypoplastic and autoimmune thrombocytopenia in most cases. In addition, significant differences in platelet function of various diseases and of different bone marrow regeneration could be demonstrated. The TP-IVBT is wellsuited for the control of platelet transfusion efficacy and may replace the in-vivo bleeding time in most cases. On the other hand, the test still shows too much of a variation and involves too much labor and cost for routine application.

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