Electronic Library of Scientific Literature


Volume 42 / No. 6 / 1995

The pathogenesis and conservative management of radiation injuries

MRC Cyclotron Unit, Hammersmith Hospital, London W12 OHS, U.K.
pp. 289-292

DNA-mediated gene transfer into mammalian cells and cancer


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia;
¹Aristotelian University of Thessaloniki, Medical Faculty, Dept. Gen. Biology, Thessaloniki, Greece

DNA-mediated introduction of genes into mammalian cells promises to be a powerful method for detecting sequences that control cell growth, confer resistance to toxic drugs, code for surface receptor proteins, or, indeed alter cell phenotype in any clearly defined way. The identification and molecular cloning of human transforming genes from neoplastic cells was enabled by the advances in existing techniques that allow DNA-gene transfer. Some methods of gene transfer which are inefficient today, should not be disregarded in the future. Every study of the genetic basis of cancer at the molecular level is an important step to the ability to influence human cancer cells and suppress their growth in vivo.
pp. 293-297

Relation of some enzyme activities and argyrophilic proteins in childhood acute lymphoblastic leukemia


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia;
¹Department of Pediatric Oncology of University Children's Hospital, Bratislava, Slovakia

In a series of 61 children with newly diagnosed acute lymphoblastic leukemia (ALL) the detection of argyrophilic proteins (AgNORs) in relation to enzymatic profile of leukemic blasts was undertaken. The method of silver staining was used to determine the number of AgNORs per nucleus of cells. The activity of 5'nucleotidase, acid phosphatase and beta-glucuronidase was assessed. The AgNOR proteins quantity varied with immunophenotype and cytochemical profile of leukemic cells. The enzyme 5'nucleotidase is known to be the marker enzyme in B-precursors ALL and acid phosphatase in T-ALL blast cells. Activity of beta-glucuronidase emerged in lymphoblasts of some cases of ALL in close relation to increased number of AgNOR proteins per nucleus of leukemic cells. Our study indicates the possible importance of beta-glucuronidase involvement and increased AgNOR quantity in the proliferative activity of leukemic cells and thus they are of value in monitoring the risk groups of leukemic patients.

Key words: Acute lymphoblastic leukemia, argyrophilic proteins, enzyme cytochemistry.
pp. 299-305

A comparison of some leucocyte differentiation markers and the adenosine deaminase and purine nucleoside phosphorylase values in B and T cell leukemias and lymphomas


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia; ¹National Cancer Institute, Bratislava, Slovakia

Peripheral blood, bone marrow and/or lymph nodes of 77 patients with T- and B-ALLs/lymphomas were characterized for their surface membrane marker profiles using flow cytometry and fluorescence microscopy. Purine metabolism enzyme activities were compared with membrane immunophenotypes.
T and B-ALLs/lymphomas subtypes were defined by the expression of surface membrane antigens detected by the monoclonal antibodies. Based on immunophenotyping we found the following characteristic marker profiles: in T-ALL - CD7, CD2, CD1, CD5, CD3, CD4, CD8, CD38, CD71; in T-NHL - CD7, CD2, CD3, CD4, CD5, CD6; in pre-B ALL - CD10, CD19, CD24, HLA-DR, CD34, in B-ALL - CD19, CD20, CD24, HLA-DR, SmIg with kappa or lambda light chains; in B-CLL - weak SmIg, kappa or lambda, CD19, CD20, CD24, CD5, HLA-DR; in B-NHL - CD19, CD20, CD22, CD24, CD5, more intensive SmIg, kappa or lambda. The cells of leukemic cases tended to have more immature phenotypes than those of lymphoma cases.
Analysis of purine metabolism enzyme activities showed that there was a correlation between the values of adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP) and various types of T- and B-ALLs/lymphomas. ADA levels in B-NHL and B-CLL were lower than those in normal cells, while ADA level in T-ALL, T-NHL, pre-B-ALL and B-ALL was higher (the average 185, 92, 73, 63 pkat.10[^-6] cells, respectively). ADA activity was significantly different between lymphocytes of control group and T-ALL (p < 0.01), between T-ALL and T-NHL (p < 0.05), between T-NHL and B-NHL (p < 0.05) and between T-ALL and B-NHL (p < 0.05). PNP activities were lower to those in normal cells.
ADA/PNP ratio increased mostly in T-ALL, less in T-NHL, pre-B-ALL and B-ALL (10.8 and 5.3 and 2.2 and 2.0, respectively). ADA/PNP ratio was significantly different between T-ALL and pre-B-ALL (p < 0.05) and between T-ALL and B-NHL (p < 0.05).

Key words: T and B acute and chronic lymphoblastic leukemias, T and B lymphomas, immunophenotype, adenosine deaminase, purine nucleoside phosphorylase.
pp. 307-312

Potential carcinogenicity of homoisoflavanoids and flavonoids from Resina sanguinis draconis (Dracaena cinnabari Balf.)


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia;
¹Institute of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic

Polarographic behavior of three homoisoflavanoids and four flavanoids isolated from the dragon's blood (Resina sanguinis draconis, Dracaena cinnabari Balf.), collected at Sokotra, was investigated in aprotic solution and an index of potential carcinogenicity tg alpha was determined. Generally, homoisoflavanoids and flavanoids were reduced in two two-electron steps, the first being reversible and the second one irreversible. The parameter tg alpha values indicated that the majority of these compounds possesses no or only marginal potential carcinogenic activity. However, it was demonstrated that some structural modifications in basic flavonoid structure lead to changed electrochemical properties and a substantial increase of derivative potential carcinogenicity.

Key words: Dracaena cinnabari Balf., Agavaceae, homoisoflavanoids, flavonoids, D.C. polarography, potential carcinogenicity.
pp. 313-316

DNA repair in Escherichia coli: The dual function of uvr genes


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia

It has been shown earlier that the starvation of E.coli for both amino-acids and thymine applied prior to UV irradiation inhibits pyrimidine dimer excision without affecting cell survival after UV irradiation. In such cells pyrimidine dimers are tolerated by a rather error-free process that depends on the activity of uvrB, recA and lexA genes. Data presented here show: (a) that the efficient toleration of unexcised dimers requires also the uvrA gene; (b) that the starvation increases the level of RecA protein about 4.7 times; (c) that the effect of starvation on subsequent pyrimidine dimer excision is reversed by a 2 h incubation in complete medium before the cells are UV irradiated. The data suggest that the uvrA, uvrB, recA, lexA dependent nonexcisional repair may be a pathway temporarily functioning in repeatedly damaged cells.

Key words: uvr-dependent nonexcision repair, E.coli, uvrA and uvrB mutants, RecA protein, UV irradiation, dimer excision.
pp. 317-323

In vitro association of mitochondrial ATP-dependent protease with mitochondrial heat-shock proteins


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia

Specific antibodies against the mitochondrial ATP-dependent protease and heat-shock proteins were used to study the association of these proteins with an abnormal bacterial protein, CRAG. It was shown that the mitochondrial ATP-dependent protease from rat liver and Zajdela hepatoma bind to the CRAG protein and that this binding was mediated through the heat-shock proteins. It was also demonstrated that the protease associated with heat-shock proteins is capable of degrading large proteins as well as small peptides in an ATP-dependent fashion. Zajdela hepatoma mitochondria, with enhanced mitochondrial proteolysis, were shown to contain more ATP-dependent protease associated with heat-shock proteins.

Key words: ATP-dependent protease, mitochondrial heat-shock proteins, intramitochondrial proteolysis.
pp. 325-329

p53 protein overexpression associates with growth patterns rather than with metastasizing in operable breast cancer


Department of Cellular and Molecular Oncology, Masaryk Memorial Cancer Institute, 656 53 Brno, Czech Republic;
¹Department of Histology, University Hospital Brno, Brno, Czech Republic;
²Department of Surgery, Masaryk Memorial Cancer Institute, Brno, Czech Republic;
³Tissue Bank, University Hospital Brno-Bohunice, Brno, Czech Republic

We have analyzed p53 protein expression in 121 primary breast cancer biopsies by immunohistochemistry using the monoclonal antibody DO-1 and polyclonal serum CM-1. p53 protein overexpression has correlated in our study with mitotic activity (p = 0.001), nuclear atypia (p = 0.002), less favorable histological type of tumor and in a lesser extent with tumor size. The inverse, but highly significant, correlation (p = 0.007) has been observed with lymph node involvement. There was also a trend for higher p53 positivity among DNA aneuploid tumors as compared with DNA diploid cases, but this was not significant.
Our study suggests that p53, at least in some patients, may not be directly involved in the process of metastatic progression in breast cancer. Preliminary data would suggest that the detection of p53 protein overexpression could be a useful additional prognostic parameter in breast cancer.

Key words: Breast cancer, p53, tumor prognosis.
pp. 331-336

Evaluation of different fixation-permeabilization methods for simultaneous detection of surface, cytoplasmic markers and DNA analysis by flow cytometry in some human hematopoietic cell lines


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia

Different fixation/permeabilization methods were investigated for their convenience for simultaneous detection of membrane marker/DNA staining or cytoplasmic marker/DNA staining by flow cytometry. Nine different methods were employed. The expression of membrane marker CD20 and cytoplasmic marker CD22 on BJAB and DAUDI cells and cytoplasmic CD3 on MOLT4 cells was measured. Optimal methods were those that combine paraformaldehyde and saponin (for membrane CD20/DNA staining and cytoplasmic CD3/DNA staining) or buffered formaldehyde-acetone (for membrane CD20/DNA staining and for cytoplasmic CD22/DNA staining). Special interest was focused on proliferation marker CD71 and nuclear antigen Ki67. We investigated CD71 in MOLT4 cells and Ki67 in MOLT4, DAUDI and BJAB cells. Both of these markers are closely associated with proliferation rate. The optimal method for the detection of Ki67/DNA staining combines paraformaldehyde with Tween 20.

Key words: Cell fixation and permeabilization, membrane and cytoplasmic markers, proliferative markers, DNA analysis, flow cytometry, hematopoietic cell lines.
pp. 337-346

Analysis of the phenotype and cellular DNA content in some leukemias by flow cytometry


Cancer Research Institute, Slovak Academy of Sciences, 812 32 Bratislava, Slovakia

Phenotype and cell cycle distribution in peripheral blood and bone marrow mononucleated cells was studied in patients with different leukemias: T-ALL, AML, CLL, CML and plasmocytoma. DNA flow cytometry with propidium iodide fluorescence was used. Differences in cell cycle between mononucleated cells from T-ALL and CML patients on one hand and normal controls on the other were seen in peripheral blood but not in bone marrow specimens. Patients with AML, CLL and plasmocytoma showed a cell cycle distribution of mononucleated cells similar to normal controls. DNA content analysis in some leukemias was discussed.

Key words: Flow cytometry, DNA analysis, cell cycle, acute and chronic leukemias.
pp. 347-352