Nucleolar immunofluorescence in bone marrow specimens of human hematological malignancies.

Leukemic cells and myeloma cells were studied in bone marrow of untreated patients with acute and chronic myeloid leukemia, chronic lymphocytic leukemia, and multiple myeloma to provide more information on the nucleolar immunofluorescence produced by antibodies no nucleoli of HeLa cells. The nucleolar immunofluorescence was mainly observed in myeloblasts of myeloid leukemias an in immature as well as mature cells of the plasmacytic or lymphocytic cell lines of multiple myeloma or chronic lymphocytic leukemia. With respect to the nucleolar immunofluorescence, both positive and negative populations of cells were noted in the specimens of all patients studied.

Staining pattern of acid non-specific esterase in lymphocytes, plasmocytes, monocytes, and leukemic cells.

The pattern of staining for ANAE in lymphocytes, plasmocytes, monocytes and leukemic cells has been studied and the effect of NaF and E 600 on this staining reaction has been investigated. In lymphocytes the transition occasionally occurring between a dot-like (T type) and granular positivity may cause difficulties when using this reaction as a marker of resting T lymphocytes. In plasmocytes as well as in myeloma cells a number of coarse granules was present in most cells and in some cases even large blocks of the reaction product occurred. NaF and E 600 inhibits already in lower concentration (2 mM and 10 micronM respectively) the strong diffuse reaction in monocytes, the effect on other cells being negligible. Using higher concentrations the inhibitory effect of NaF (20 mM) was more pronounced in plasmocytes in comparison with T lymphocytes which on the other hand, were, in general more sensitive to E 600 (10 mM). Partially diffuse or dense granular pattern of ANAE was found in a number of hairy cells. "Lymphoblastic" leukemias in children most frequently displayed negative or fine granular reaction in a varying number of blasts and similar findings were noted in the majority of cytochemically undifferentiated leukemias in adults. In some of them, however, a similar pattern of ANAE as in myeloblastic leukemias with dispersed or partial diffuse positivity was observed. The significance of these findings is briefly discussed.

The blood volume and splenic red cell pool in hairy cell leukemia.

Investigating the splenic red cell pool in 7 patients with hairy cell leukemia we have found unusually high values of retained erythrocytes (means 527 ml, SD 264) with a range from 300 to 998 ml. This amount represents on average 26% of the total red cell mass. The blood volume in these patients ws 26% greater than predicted by NADLER et al. Following splenectomy, the hemoglobin improved with 49.3% (SD 29.1).

Intramitochondrial rodlet-like inclusions in human leukemic lymphocytes.

Some mitochondria in leukemic cells in two of nine investigated patients with chronic lymphocytic leukemia and in one patient with morphologically less usual form of this disease contained unusual rodlet-like inclusion bodies. These structures were always present in the matrix of frequently dilatated intercristal spaces. The width of rodlet-like inclusions ranged between 10 and 20 nm, their maximal length was 114 nm. All observed rodlet-like inclusion bodies appeared to be composed of fine filaments 2...3 nm in width and resembled the DNA containing structures in mitochondria of few other cell types. In mitochondria of leukemic cells, however, these inclusions represent a further structural abnormality.

The influence of serum content in the medium on the survival of human bone marrow in suspension in vitro.

Samples of human bone marrow cells from patients with various diseases were cultivated in vitro by means of a simple stationary suspension method. Medium Eagle MEM with the addition of allogeneic serum and borosilicate glass were used. The cells survived significantly longer in samples with 50 per cent of serum than in samples with 30 per cent of serum only. Monocytes showed the longest survival (max. 95 days) in cultures with 50 per cent of serum. Myelocytes and segmented neutrophils as well as normoblasts survived till 45th day and plasmocytes till 66th day. Mitoses in monocytes were found till 50th day. Moreover so called "satellitosis" of plasmacytes around a macrophage was observed in cases with reactive plasmacytosis.

Peroxidase and pseudoperoxidase reactions in relation to sudanophilia.

A selective staining of hemoglobin in erythroid cell series was achieved by use of Sudan Black B (modified method of Sheehan and Storey) if optimal amount of hydrogen peroxide was added to the staining mixture. The effect of some inhibitory agents (KCN, wet heat, pH) on this staining as well as on the Lepehne's pseudoperoxidase reaction for hemoglobin was similar. Both reactions were more resistant to these factors than the peroxidase reactions and sudanophilia in granulocytes in which both could be blocked by the pretreatment with absolute methanol. Moreover the effect of some extraction procedures for lipids on both myeloperoxidase reactions and sudanophilia was investigated. The results support the view that the sudanophilia in granulocytes is due to their peroxidase activity and for the staining of hemoglobin by use of Sudan Black B with H2O2 its pseudoperoxidase activity is responsible. In addition the effect of the substitution of phenolphosphate by dihydroxybenzenes on granulocyte sudanophilia is reported.