Characteristics of the silver-stained nucleoli of tumor cells from the pleural fluid of lung cancer patients

Tumor cell nucleoli obtained from pleural effusions of 26 patients with different morphologic types of lung cancer were evaluated by silver staining. Distinct heterogeneity of tumor cell populations, with regard to the number of nucleoli as well as their functional activity in respect to ribosomal RNA synthesis, were shown to be the most common feature of all the tumors studied, regardless of their morphologic variants. One likely cause of heterogeneity in Ag nucleolar organized region (NOR) pattern of tumor cells may be due to chromosomal losses and gains from the karyotypes of acrocentric chromosomes with active NORs. Another possible cause for heterogeneity in nucleolar activity might be due to different reactions of tumor cells towards some humoral and cellular factors of pleural fluid including T-lymphocytes.     

Stereoscopic back-scattered electron imaging of silver-stained proteins in nucleoli

By using simultaneously the AgNOR silver staining method, back-scattered electron imaging mode and stereo-tilt in scanning electron microscopy (SEM), it is possible to observe the nucleus through the cell surface, the nucleolus, and the tri-dimensional distribution of the AgNOR-associated acidic proteins. In C3H10T1:2 cells and their 7-12-dimethylbenz--anthracene-treated transformants, the staining demonstrates several intranucleolar silver-staining granules (SSG), surrounded by a weakly staining region. The SSG may represent the fibrillar center (FC) and the weakly staining region, the fibrillar dense component (FD). This component can link several SSG together to form a rope-like structure. In cells with no visible nucleolus and inactive nucleolar organizer regions (NORs) the silver-staining granules are less numerous, close together and the presumed fibrillar dense components are not visible. The SSG are located more peripheraly, and the weakly staining region and the rope-like structure are less prominent in control cell nucleoli than in transformed cells with a comparatively high rate of RNA synthesis.     

Change in morphometric parameters in silver-stained nucleoli from hepatocytes of rats with liver cirrhosis and during rehabilitation from it]

Silver-stained nucleoli of rat hepatocytes were studied in norm, in liver cirrhosis produced by CCl4 poisoning and after cessation of the poisoning. Morphometric parameters of nucleoli were measured using a Videotest computer image analyser. Under cirrhosis the mean number of nucleoli per nucleus was determined to exceed their normal number by 1.27 times. The total volume of nucleoli in the nucleus also exceeded the normal level (by 1.15 times). 3 months after the end of CCl4-poisoning, these parameters decreased almost to normal values. A statistically significant correlation was revealed between the number of nucleoli and their total volume (0.881). Changes of the parameters also correlated with the total protein content in the hepatocytes. Possible reasons for this correlation are discussed. The ratio of the number of chromosomal NORs to the mean number of nucleoli in the nucleus is proposed to be used as a feature for comparative analysis of functional status of nucleoli in the nuclei of different ploidy and in cells of different animal species.     

Measurement of the diameters of nucleoli by the template method

The method of measuring the nucleoli diameters in the light microscope is described. The nucleoli are compared to two standard templates followed by calculation of the means value and the standard deviation by means of special formulas. The scale marks of ocular micrometer or of special ocular scale can be used as the templates. The use of templates makes the measuring of nucleoli diameters 3-3.3 times as quick as the screw ocular micrometer, and with good correlation between the results obtained with both methods, too.     

Experience with using aclarubicin in the treatment of acute leukemia and blast crisis of chronic myeloid leukemia]

The clinical efficacy of aclarubicin, an anthracycline antibiotic, was studied in 48 patients with leukemia. The antibiotic was used in the following combinations with cytarabine: "7 + 7", "5 + 5" and "7 & 3". A complete remission was stated in 14 (42.4 per cent) out of 33 patients with acute nonlymphoid leukemia, 6 (43 per cent) out of the 14 patients having relapses. The combined therapy was effective in 4 out of 5 pre-resistant patients. The "7 + 3" scheme was the most beneficial. The most common adverse reactions were nausea and vomiting.     

Hydrocortisone in culture protects the blast cells in acute myeloblastic leukemia from the lethal effects of cytosine arabinoside

The blast cells in acute myeloblastic leukemia (AML) respond to many of the same regulatory mechanisms that control normal hemopoiesis. These include the growth factors that bind to membrane receptors and steroid hormones or vitamins that have intracellular receptors. We report the effects in culture of the steroid glucocorticoid hydrocortisone on freshly explanted AML blasts from patients and on two continuous AML cell lines. Only small changes in clonogenic cell numbers in suspension cultures were seen in the presence of hydrocortisone. The most striking effect of the hormone was on the sensitivity of blasts cells to cytosine arabinoside (ara-C). In contrast to the response of AML blast cells to retinoic acid, a ligand for intracellular steroid receptors that sensitizes some blast populations to ara-C, hydrocortisone reduced the toxic effects of the drug. The protective action of hydrocortisone was not mediated through the cell cycle since exposure of blasts to hydrocortisone did not affect the percentage of cells in DNA synthesis as measured with the tritiated thymidine (3HTdR) "suicide" technique. The hydrocortisone effect could be demonstrated using a pulse (20 min) exposure protocol. Blasts pulsed with increasing specific activities of 3HTdR showed the usual response pattern with an initial loss in plating efficiency to about 50% of control, followed by a plateau, regardless of whether the cells had been exposed to hydrocortisone. Control blasts exposed to increasing ara-C concentrations gave very similar dose-response curves; in striking contrast, blast cells cultured in hydrocortisone, then pulsed with ara-C did not lose colony-forming ability even though the same population was sensitive to 3HTdR. The hydrocortisone effect was dose and time related; protection from ara-C increased from 10(-8) to 10(-5) M and was seen after 4 hr exposure but required 8 hr to reach a maximum. We conclude that hydrocortisone can protect blasts from the lethal effects of ara-C even while the cells are in active DNA synthesis.     

Blood lymphocyte volumes and diameters in patients with chronic lymphocytic leukemia and normal controls

The electronic modal lymphocyte volumes of 151 patients with chronic lymphocytic leukemia (CLL) and 305 normal controls were determined by the hydrodynamically focused multi-channel Coulter TF analyser. The mean volumes of the normally distributed groups were 166 +/- 19.3 (range 126-216) fl in patients with CLL and 206 +/- 14.4 (range 126 +/- 246) fl in normal controls. The calculated cell diameters were 6.8 (6.2-7.4) micron and 7.3 (6.8-7.8) micron respectively. Our data do not support previous reports about relations between cell size and clinical stages of the Rai and Binet classifications.     

Characteristics of DNA replication of the chromosomes of the bone marrow cells in patients with acute leukemia]

A study of the late DNA replication pattern in chromosomes of human acute leucaemia cells revealed a significant diffrence from control. Chromosomes, 2,3 and 4-5 of the acute leucaemia cells finish their DNA replication earlier, and chromosomes 1, 13-15 and 16 later, compared to the control chromosomes. The difference in the pattern of DNA replication between analogous chromosomes of acute leucaemia and donor cells was associated with the discovery of large late-replicating chromatin blocks in the pericentromeric regions of leucaemia cell chromosomes. Some relationship is suggested between the pattern of pericentromeric heterochromatin DNA replication and cell differentiation.     

Risk factors occurring in various types of acute leukemia

The present investigations demonstrate that as far as the genesis of non-lymphatic leukemias is concerned environmental factors may be assumed as an aetiological factor to an increasing extent. Medicaments, particularly cytostatics, exogenous noxious occupational agents and infectious-inflammatory diseases might be significant for developing ANLL, whereas virus infection, immunizations, and acclimatization to specific conditions of climate may be rather attributed as risk factors for ALL. A tendency to deviations from the blood picture ( lymphomonocytosis , leukocytopenia, toxic granulation of neutrophilic granulocytes) can be increasingly observed in workers engaged in chemical industry, with these changes, being still reversible. A permanent effect of exogenous toxic agents, however, will produce a serious damage of the bone-marrow. Further investigations are provided for elucidating the function of different haemopoietic cell systems, particularly the immune status, in workers of those enterprises where an exposure to noxious substances can be found.     

In vitro autologous blast cell sensitization as immunotherapy in acute non-lymphocytic leukemia

Summary Remission mononuclear cells incubated in vitro for 96 h with autologous stored blast cells were reinfused IV on two occasions as adjuvant maintenance therapy. The procedures were well tolerated, but in vitro response to the blast cells was negligible.The present mean duration of complete remission (15.7 months) appears to be similar to that of 28 patients (12.4 months) treated at the same time with chemotherapy alone by the same physicians.     

Autonomous growth potential of leukemia blast cells is associated with poor prognosis in human acute leukemias

Background

Anaplastic lymphoma kinase-positive diffuse large B-cell lymphoma (ALK-DLBCL) is a rare lymphoma with several clinicopathological differences from ALK-positive anaplastic large cell lymphoma (ALCL). The latest WHO classification of lymphomas recognizes ALK-DLBCL as a separate entity.

Methods

A comprehensive comparison was made between the clinical and pathological features of the 4 cases reported and those found in an extensive literature search using MEDLINE through December 2008.

Results

In our series, three cases were adults and one was pediatric. Two cases had primary extranodal disease (multifocal bone and right nasal fossa). Stages were I (n = 1), II (n = 1), III (n = 1) and IV (n = 1). Two cases had increased LDH levels and three reported B symptoms. IPI scores were 0 (n = 1), 2 (n = 2) and 3 (n = 1). All cases exhibited plasmablastic morphology. By immunohistochemistry, cases were positive for cytoplasmic ALK, MUM1, CD45, and EMA; they marked negative for CD3, CD30 and CD20. Studies for EBV and HHV-8 were negative. The survival for the patients with stage I, II, III and IV were 13, 62, 72 and 11 months, respectively.

Conclusion

ALK-DLBCL is a distinct variant of DLBCL with plasmacytic differentiation, which is characterized by a bimodal age incidence curve, primarily nodal involvement, plasmablastic morphology, lack of expression of CD20, aggressive behavior and poor response to standard therapies, although some cases can have prolonged survival as the cases reported in this study. ALK-DLBCL does not seem associated to immunosuppression or the presence of EBV or HHV8. Further prospective studies are needed to optimize therapies for this entity.     

The immunophenotypic and cytogenetic characteristics of the blast cells in subvariants of acute myeloid leukemia

The results of study of morpho-cytochemical peculiarities, antigenic profile and peripheric blood and/or bone marrow blast cell karyotype of 21 patients with acute myeloid leukemia are presented. Hemopoietic cell immunophenotyping was carried out with the use of cytofluorometer FACScan and chromosome cytogenetic analysis with the use of analyzer "Metascan". It has been shown that in the AML M1 blast cell plasmatic membrane carries pan-myeloid CD33 and CD13 antigens, the last having high density of expression, and the CD38 antigen, which is a myeloid cell-precursor marker. In these patients tetraploidy, being the testimony of karyotype change, has been ascertained. It has been found out that the AML M2 blasts, except pan-myeloid antigens, express the cell proliferation CD71 marker. Blast cell karyotype peculiarities typical for this leukemia sub-variant have been revealed. In patients with the AML M4 in 3 of 6 cases an anomalous karyotype has been found. It has been also shown that the CD14 antigen, and rather its percentage in blast total population, is the differential-diagnostic criterion for the AML M4 and M5.     

Bleomycin-reactive iron in patients with acute non-lymphocytic leukemia.

Bleomycin-reactive iron was detected in the sera of six out of nine adults undergoing intensive chemotherapy for acute non-lymphocytic leukemia. In these individuals the corresponding transferrin saturation ranged from 96% to 113% and the serum ferritin from 775 to 9975 micrograms/l. Nontransferrin-bound iron has been postulated to be a factor in organ toxicity in iron overload conditions such as beta thalassemia and hereditary hemochromatosis by facilitating the production of tissue-damaging free radicals. We propose that bleomycin-reactive iron should be considered as a possible factor in organ dysfunction seen with intensive cancer chemotherapy.