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H Klamová

Publications and source records attributed to H Klamová.

At least 19 recordsLinked to original sources
PubMed2006

A karyometric note on nucleoli in human early granulocytic precursors.

The diameter of nucleoli was measured in human bone marrow early granulocytic precursors after visualization by a simple cytochemical method for demonstration of RNA. Such method facilitated to clearly see nucleolar bodies without perinucleolar chromatin, including those of micronucleoli. The bone marrow of patients suffering from chronic myeloid leukaemia (untreated with cytostatics) provided a satisfactory number of both myeloblasts and promyelocytes for nucleolar measurements because of prevailing granulopoiesis. The direct nucleolar measurement was carried out on digitized and processed images on the screen at magnification 4,300x. It seems to be likely that the nucleolar size is directly related to the number of nucleoli per cell. The largest nucleoli were present in both myeloblasts and promyelocytes that possessed a single nucleolus. In contrast, the nucleolar diameter was significantly smaller in cells with multiple nucleoli. However, in cells with small multiple nucleoli, one of them was always larger and dominant with a large number of AgNORs. Such large nucleoli are possibly visible in specimens stained with panoptic procedures or methods staining nuclear chromatin or DNA. It should also be mentioned that both myeloblasts and promyelocytes mostly possessed two nucleoli with the mean diameter close to 1.5 microm. The incidence of early granulocytic precursors classified according to the nucleolar number and size strongly suggested that the various nucleolar number and nucleolar size in these cells might be related to the different stage of the cell cycle and might also explain their heterogeneity.

Cell Nucleolus↗
PubMed2006

Immunological profiles of patients with chronic myeloid leukaemia. I. State before the start of treatment.

In view of the increasing interest in the immunotherapy of CML it seems highly desirable to broaden the present knowledge on the immune reactivity of CML patients. A group of 24 patients and 24 healthy controls were studied for the total of 15 immunological parameters, including the prevalence of antibodies against human herpesviruses and papillomaviruses. To clearly discriminate between changes associated with the disease and those induced by the therapy, all patients were enrolled prior to the start of any anti-leukaemic therapy. Statistically significant differences between patients and controls were found in the levels of IgA, C4 component of complement, CRP and IL-6, the production of Th1 cytokines in stimulated CD3 cells and the E. coli stimulatory index. The analysis of the interrelationship between the results obtained in the individual patients presented some unexpected findings, such as the lack of correlation between the CRP and IL-6 levels. It will be the purpose of a follow-up to determine whether and how the immune status of the patients prior to the treatment correlates with their response to therapy and how the individual immunological profiles change in the course of the disease. These observations will be utilized in the future immunotherapeutic studies to constitute the vaccine- and placebo-treated groups.

Adult↗
PubMed2005

Nucleoli in human early erythroblasts (K2, K1, K1/2 cells).

Human early erythroid precursors classified according to the nuclear size were studied to provide information on nucleoli in these cells using simple cytochemical procedures for demonstration of RNA and proteins of silver-stained nucleolar organizers. K2 cells with nuclear diameter larger than 13 microm and K1 cells with nuclear diameter larger than 9 microm corresponding to proerythroblasts and macroblasts (large basophilic erythroblasts) mostly possessed large irregularly shaped nucleoli with multiple fibrillar centres representing "active nucleoli". K1/2 cells with nuclear diameter smaller than 9 microm corresponding to small basophilic erythroblasts were usually characterized by the presence of micronucleoli representing "inactive nucleolar types". On the other hand, a few K1/2 cells contained large nucleoli with multiple fibrillar centres similar to those present in K2 cells and thus appeared as "microproerythroblasts". The nucleolar asynchrony expressed by the presence of large irregularly shaped nucleoli with multiple nucleoli (active nucleoli) and ring-shaped nucleoli (resting nucleoli) in one and the same nucleus of K2 or K1 cells was not exceptional and might reflect a larger resistance of these cells to negative factors influencing the erythropoiesis. The intranucleolar translocation of silver-stained nucleolus organized regions was noted in K2 cells and might indicate the premature aging of these cells without further differentiation. More studies, however, are required in this direction.

Cell Differentiation↗
PubMed2005

Intranucleolar translocation of AgNORs in early granulocytic precursors in chronic myeloid leukaemia and K 562 cells.

The present study was undertaken to provide missing information on the distribution of AgNORs in large nucleoli of human leukaemic early granulocytic precursors in vivo as well as in vitro. In vivo, the distribution of AgNORs was studied in early granulocytic precursors of patients suffering from chronic myeloid leukaemia who were both untreated and treated with imatinib mesylate. AgNORs were visualized by silver reaction under conditions which facilitated to see their distribution by light microscopy. In vitro, the distribution of AgNORs was studied in proliferating and ageing K 562 cells which originated from chronic myeloid leukaemia. In vitro, the ageing of K 562 cells produced intranucleolar translocation of AgNORs to the nucleolar periphery. Such translocation was also observed in some leukaemic early granulocytic precursors in vivo, e.g. in bone marrow myeloblasts and promyelocytes of leukaemic patients. As was expected, the intranucleolar translocation of AgNORs in early granulocytic precursors was more frequent in patients treated with the cytostatic therapy--imanitib mesylate. The abovementioned findings suggest that myeloblasts and promyelocytes with AgNORs translocated to the periphery of large nucleoli might be in the ageing state, similarly as blastic cells of leukaemic myeloid origin (K 562 cells) in ageing cultures. Thus, the translocation of AgNORs might be a useful marker of premature ageing in the future and might contribute to the evaluation of the single cell state under various experimental as well as clinical conditions. However, more clinically oriented studies are required in this direction.

Cell Line, Tumor↗
PubMed2004

[Imatinib mesylate (Glivec) in treatment of chronic phase chronic myeloid leukemia].

The new knowledge in molecular biology and pathophysiology of chronic myeloid leukemia enabled the development of imatinib mesylate (Glivec, formerly STI571). Imatinib potently inhibits several protein tyrosine kinases, including BCR-ABL, c-Kit, and PDGF receptor. Imatinib blocks the phosphorylation of downstream target proteins and interrupts the malignant transformation leading to the development of CML. Phase I and II studies demonstrated that imatinib is highly effective and well tolerated in all phase of CML. We got our experience with imatinib on more than two-year monitoring 34 patients within the Expanded Access Study CST1571 0113. Imatinib 400 mg/d was administered orally to 10 women and 24 men in median age of 53 years (22-70) who were hematologically (n = 9) or cytogenetically (n = 13) resistant, cytogenetically refractory (n = 3) or intolerant (n = 9) to interferon alpha. The median follow-up time was 97.5 weeks (23-115), the median time from CML diagnosis to the start of the study was 32.3 months (6-140.5). Complete hematologic response was achieved in 33 of 34 (97%) pts, total major cytogenetic response (complete plus major) in 21 of 33 (63%) pts. Cytogenetic relapse was observed in 2 of 33 pts (6%), cytogenetic progression in 4 (12%) pts. Non-hematologic toxicity was mild (grade 1 or 2) and no patient was excluded from the study due to it. Hematological toxicity grade 3 limited dose of imatinib in 26% of patients and probably caused lower rate of cytogenetic responses in heavy pretreated patients. Both quantitative RT-PCR methods (competitive RT-PCR and real-time RT-PCR Light-Cycler) were found useful to monitor patients with CML on imatinib therapy. Our results confirmed high efficacy and safety of imatinib in late-chronic phase CML patients failing prior interferon therapy. The lower incidence of hematological toxicity and higher rate of cytogenetic responses in patients treated with imatinib in early-chronic phase CML justify according to our opinion the recommendation to administer imatinib early after the diagnosis of CML in patients who are not indicated for allogeneic transplantation.

Adult↗
PubMed2004

[Imatinib--a new perspective in the treatment of tumors].

Among novel promising approaches to anticancer therapy belongs the targeting inhibition of signal transduction. This review outlines present-day experiences with imatinib (Glivec), a potent inhibitor of the tyrosine kinases bcr-abl, c-kit and platelet-derived growth factor receptor kinase. Due to inhibition of bcr-abl tyroxine kinase, imatinib has rapidly become the standard therapy for chronic myelocytic leukemia; inhibition of c-kit receptor explains its effectivity in the treatment of patients with gastrointestinal stromal tumors. Another known target of imatinib is tyrosine kinase of PDGFR, which is activated in numerous malignancies, particularly in dermatofibrosarcoma protuberans. Discovery of the novel fusion gene in hypereosinophilic syndrome (FIPILI-PFGFRA, whose product is an imatinib sensitive protein kinase) permitted to treat successfully this event. Possible combination of imatinib with conventional chemotherapeutic drugs and other key signal transduction inhibitors are mentioned.

Antineoplastic Agents↗
PubMed2002

[Use of photodynamic therapy for elimination of residual leukemic cells in autologous transplants of hematopoietic progenitor cells].

BACKGROUND: The increasing use of autologous hematopoietic cell support in various malignancies including leukemia and lymphoma currently bears the problem of tumor contamination of the graft with tumor cells which after re-infusion contribute to the disease relapses. It is therefore desirable to eradicate the cancer cell fraction of the graft without causing damage to the normal stem cell fraction. The purging processes based on photodynamic treatments appear to be perspective means for this purpose. METHODS AND RESULTS: We investigated the effects of 5-aminolevulinic acid (ALA)--based photodynamic treatment (ALA-PDT) on the proliferation of human leukemia cell lines HL60 (promyelocytic leukemia), ML2 (myelomonocytic leukemia) and HEL (erythroleukemia) by 3H-thymidine incorporation into the cell DNA, on the viability of cell lines HL60, HEL, DAUDI (B-cell leukemia) and JURKAT (T-cell lymphoma) as well as of blast cells of acute myeloid leukemia (AML) patients by flow cytometry-propidium iodide assay, and on the clonogenic activities of normal human bone marrow cells by in vitro cloning assays. Under the conditions used (treatment with 1 mM ALA for 4 h at 37 degrees C followed by exposure to blue light dose of 18 J/cm2) the number of proliferating HL60 cells was reduced by 2.4 logs, of ML2 cells by 3.2 logs and of HEL cells by 1 log. From the mononuclear cell preparations of AML patients the blast cells were substantially reduced in eight out of ten patients. The clonogenic activities of normal bone marrow progenitor cells were largely spared: 52.5 +/- 8.9% of colony-forming units--granulocytes macrophages (CFU-GM), and 48.6 +/- 9.7% burst forming units--erythrocytes (BFU-E). CONCLUSIONS: ALA-PDT appears to be usable principle for the depletion of residual leukemic cells from autologous transplants.

Aminolevulinic Acid↗
PubMed2000

[Gene expression in white blood cells in chronic myeloid leukemia].

BACKGROUND: The new technologies that have the DNA laboratory over recent years and the general progress in knowledge of the human genome, have allowed the simultaneous observation of the activity of a large number of genes. Chronic myeloid leukemia is characterized with abnormal tyrosine kinase activity of the fused bcr/abl gene, which is most often product of translocation between chromosomes 9 an 22. It is as yet unknown whether this is the only and sufficient cause of the disease, or whether other supporting and co-active abnormalities exist. It is also not yet clear whether an increase of proliferating activity or reduced programmed cell death plays the dominant role. The aim of this study was to make further steps in resolving the question as to which of these hypotheses fits better. METHODS AND RESULTS: Membrane macroarrays (Clontech 7742-1: Human Cancer cDNA Expression Array with 588 gene probes) were used throughout the study, on which cDNA reverse-transcribed from total RNA in turn isolated from peripheral white blood cells and labelled with 32P was hybridized. Cells obtained from 5 patients with confirmed diagnoses by cytogenetic and molecular (bcr/abl) analyses, but who had not yet been treated by chemotherapy, were the source of the material. In some cases mononuclears and granulocytes were also isolated by Ficoll-Paque centrifugation. Radioactivity was detected by autoradiography or by a Phosphorimager (Fujifilm FLA-2000). Comparison with normal gene expression (healthy donor) was made by subtraction using Clontech AtlaImage 1.5 software. Although changes of expression of identical genes were not observed in all of patients examined, the majority of them were concordant. Values at least double those of the controls applied to the activity of c-jun N-terminal kinase, MMP-8, MMP-9, integrin alpha E, integrin beta and PDGF, whereas the expression of ZAP-70, IRF1, MCL-1, STAT 5B, RARA, CDC25B, RPSA, TNFR decreased. Increases of PCNA, MMP-17, CD59, rho G, CRAF1 and PIG7 or decreases of notch, caspase 8, caspase 4, interleukin 6 receptor, rho B and TIMP1 were observed only in some cell samples. CONCLUSIONS: It seems that some maturation processes and transmembrane signalling are blocked, as well as the effectors of apoptosis. On the other hand, the reduced activity of ZAP-70, IRF1 and MCL-1 also indicated that proliferation breaks were weakened. The involvement of both processes-released replication and ineffective apoptosis--was evident; the problem of bcr/abl gene fusion being the necessary first and sufficient step on the way towards developing chronic myeloid leukemia, however, remained unresolved.

Fusion Proteins, bcr-abl↗
PubMed2000

[Selective photodynamic destruction of leukemic cells].

BACKGROUND: Residual leukemic cells if present in autologous bone marrow grafts or CD34+ concentrates obtained from peripheral blood may increase the risk of relapse after autotransplantation. We are presenting the employment of a new method which was introduced into the photodynamic therapy, namely enhancement of synthesis of the photosensitizing compound, protoporphyrin IX, in cancer cells, following application of its metabolic precursor, 5-aminolevulinic acid, for the specific destruction of leukemic cells. METHODS AND RESULTS: By determining cell viability using tetrazolium salt reduction (MTT), by flow cytometrypropidium iodide assay and by determining cell proliferation using bromodeoxyuridine incorporation we studied the effect of photodynamic therapy based on the application of 5-aminolevulinic acid on the cells of leukemic cell lines HL60 (human promyelocytic leukemia), HEL (erythroleukemia), DAUDI (B-cell leukemia), JURKAT (T-cell lymphoma), blast cells of patients with acute myelogenous leukemia as well as on normal lymphocytes and normal human bone marrow progenitors. In in vitro experiments photodynamic therapy based on an administration of 5-aminolevulinic acid (1 mM, 4 h, 18 J/cm2) lowered the number of viable leukemic cells by over 2 orders (with the exception of HEL cells) and eliminated blast cells in mononuclear cell preparations of six out of seven patients with acute myelogenous leukemia. On the other hand the viability of normal resting lymphocytes was little affected by photodynamic therapy (number of necrotic cells increased from 6 to 11%) and also the clonogenic activity of the progenitor cells of normal bone marrows did not decrease substantially (CFU-GM to 60% and BFU-E to 55% of the original activity). CONCLUSIONS: Photodynamic therapy based on the application of 5-aminolevulinic acid is a perspective method for the specific destruction of leukemic cells in autologous transplants.

Aminolevulinic Acid↗
PubMed1999

Occurrence of cytokine receptors on different lymphoid leukaemic cells.

We have studied the expression of cytokine receptors CD25 (IL-2R alpha, 55 kD), CD116 (hGM-CSFR, 145 kD), CD117 (CSFR, 145 kD), CD120a (TNFR, 55 kD), CD120b (TNFR, 75 kD), CD121a (IL-1R, type I, 80 kD), CDw123 (IL-3R), CD124 (IL-4R, 140 kD), CD126 (IL-6R, 80 kD), CD127 (IL-7R, 75 kD), CDw128 (IL-8R), CD130 (gp130 subunit), CDw131 (common beta), CD132 (IL-2Rgamma), CD134 (OX40) and also CD95 (Fas antigen) on the lymphoid leukaemic cells. Cells from peripheral blood or bone marrow of 24 patients with disorders in lymphoid lineage mostly included acute lymphoid leukaemias (with a high leukocyte count and percentage of blasts) were analysed for the expression of surface membrane molecules by the immunofluorescence method evaluated by flow cytometry. The findings indicate that some monoclonal antibodies have a reactivity against cytokine receptors of pathological cells in individual cases, but with very variable qualitative and quantitative expression (number copies/cell). The lymphoid leukaemic cells demonstrate unique cytokine receptor profiles, which reveal the great diversity of immunophenotypes within the main functional characterisation of T and B lymphoproliferative malignancies.

Adolescent↗
PubMed1999

A note on nucleoli in granulocytic precursors of the granulopoietic proliferating compartment in patients suffering from the chronic phase of chronic myeloid leukemia treated with two different drugs with different mode of action.

The incidence of main nucleolar types in granulocytic precursors was studied in the granulopoietic proliferating compartment (GPC) of patients suffering from chronic phase of the chronic myeloid leukemia (CML) who were treated by the widely used therapy with two different drugs with different mode of action - hydroxyurea (HU) and interferon alpha (IFN-alpha). In comparison with IFN early stages of GPC, i.e. myeloblasts and promyelocytes in patients treated with HU possessed more frequently micronucleoli which are known to reflect the direct as well as indirect inhibition of the nucleolar biosynthetic activities. On the other hand, the incidence of micronucleoli in these cells of a small percentage of patients treated with IFN also reached the average values of these nucleoli which were noted in patients treated with HU.

Cell Nucleolus↗
PubMed1998

[Interferon alpha--the drug of choice for patients with chronic myeloid leukemia].

BACKGROUND: The conventional, or standard, treatment of chronic myeloid leukaemia (CML) with hydroxyurea and busulfan has no marked influence on its course or duration. Interferon (IFN) alpha administration has, on the other hand, been shown to induce not only a haematological but also cytogenetic response, i.e. partial or complete bone marrow repopulation by Ph-negative cells. This has triggered studies comparing IFN and conventional chemotherapy. The present work had the purpose to gain experience with IFN alpha treatment and compare the results with hydroxyurea and busulfan treatment, in the chronic phase of CML. METHODS AND RESULTS: Therapeutic results obtained in 30 patients given IFN alpha and 30 others given conventional chemotherapy were evaluated retrospectively. In spite of the short time of IFN administration (4-27 months), significantly more complete haematological responses (83%) were observed in this than the conventional chemotherapy group (57%). Cytogenetic responses were achieved in 36%, complete cytogenetic remission in 20% of IFN-treated patients. Conventional chemotherapy produced no cytogenetic effect. CONCLUSION: The results obtained confirm the value and efficacy of IFN-alpha treatment in CML patients, especially if it is started early and the dose is effective. Regular cytogenetic monitoring is necessary. Longer follow-up of the patients will be necessary for evaluation of the IFN effect on the length of their survival.

Adult↗
PubMed1998

The nucleolar functional asynchrony--imbalance--in the granulopoietic proliferating compartment of patients suffering from chronic myeloid leukemia.

Nucleoli were studied in all stages of the granulopoietic proliferating compartment in the bone marrow of patients suffering from chronic myeloid leukemia to provide an information on the incidence of the nucleolar functional asynchrony (imbalance) in these cells. The nucleolar functional asynchrony is morphologically expressed by the presence of "active" large nucleoli with a relatively uniform distribution of ribonucleic acid (RNA) and "resting" ring shaped nucleoli with RNA only in their peripheral part in one and the same cell. This phenomenon was noted in a small but constant percentage of myeloblasts and decreased in myelocytes regardless of the phase of the disease and therapy. In addition, the nucleolar functional asynchrony was also noted in all stages of the granulopoietic proliferating compartment of control not-leukemic persons.

Bone Marrow Cells↗
PubMed1998

Selective destruction of leukaemic cells by photo-activation of 5-aminolaevulinic acid-induced protoporphyrin-IX.

The effect of 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT) on the viability and proliferation of leukaemia/lymphoma cells as well as normal human lymphocytes has been investigated by flow cytometry-propidium iodide assay (FC-PI), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and bromodeoxyuridine (BrdU) incorporation and on clonogenic activity of normal human bone marrow progenitor cells by clonogenic methods. ALA-PDT (1 mM 5-ALA, 4 h, 18 J cm-2) reduces the number and/or suppressed proliferation of leukaemic cells of promyelocytic (HL60), B-cell-derived (DAUDI) and T-cell-derived (JURKAT) cell lines by 2 logs and that of the HEL erythroleukaemia cells by 77%. The effect of ALA-PDT on quiescent human lymphocytes is small (85% viable cells after ALA-PDT). The proliferation of lymphocytes subjected to ALA-PDT and induced with phytohaemagglutinin (PHA) decreases by 75% as compared to the untreated control. For normal human bone marrow progenitors, 58% of colony-forming units-granulocytes-macrophages (CFU-GM) and 55% burst-forming units-erythrocytes (BFU-E) activities are preserved.

Aminolevulinic Acid↗
PubMed1998

P230 BCR/ABL protein may be associated with an acute leukaemia phenotype.

The BCR/ABL rearrangement, the molecular hallmark of chronic myelogenous leukaemia (CML), is rare in acute myeloid leukaemia (AML), being detected in approximately 1% of cases. In the vast majority of CML cases the breakpoint on chromosome 22 falls in the so-called major breakpoint cluster region of the BCR gene. Only a few cases of CML with breakpoint in the minor or in the micro bcr region have so far been reported. The micro breakpoint position has been associated mainly with a mild form of CML, defined as Philadelphia chromosome-positive chronic neutrophilic leukaemia (Ph-positive CNL). Using reverse transcription-polymerase chain reaction (RT-PCR) we report a patient with an acute myeloid leukaemia phenotype at diagnosis who showed a BCR/ABL rearrangement with a breakpoint located in the micro bcr region (e19a2 junction). Cytogenetic analysis showed a progression of the malignant clone, finally leading to cells with two Ph chromosomes, trisomy 8, isochromosome 17q and deletion of the long arms of chromosome 7. The findings of chromosomal changes point to a possibility of blast crisis of CML with a clinically silent chronic phase. Immunoprecipitation and auto-phosphorylation assay revealed the expression, by the patient's blast cells, of an abnormal P230 BCR/ABL protein, which showed for the first time that this protein was constitutively activated in primary cells from patients. This finding may contribute to the understanding of the role of the BCR/ABL rearrangements in determining different leukaemia phenotypes ranging from acute lymphoid and myeloid leukaemias to mild chronic neutrophilic leukaemias.

Acute Disease↗