Significance of Improved Cytochemical Methods for Hemoprotein Enzymes in Diagnosis and Classification of Leukemia

Hanker, Jacob S.; Yates, Peggy E.; Ambrose, Wallace W.; Moore, Joseph O.; László, John; Starkweather, William H.; Giammara, B.

doi:10.1007/978-1-4613-2823-0_3

Cited by 3 publications

(2 citation statements)

References 35 publications

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“…Hanker et a1 (14) claimed that their cytochemical technique can discriminate between peroxidase of neutrophils, eosinophils and basophils by differences in colour, i.e., neutrophil granules are grey-black, those of eosinophil are red-brown while basophil granules are violet-black.…”

Section: Methodsmentioning

confidence: 99%

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Cytochemical demonstration of ‘platelet’ peroxidase at the light microscope level

Khalaf¹,

Hayhoe²

1987

European J of Haematology

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A reliable and reproducible modified method was developed for the cytochemical demonstration of the so-called 'platelet' peroxidase at the light microscopy (LM) level.Interestingly, not only normal platelets and megakaryocytes showed the peroxidase activity, but also normal lymphocytes. As with the ultrastructural platelet peroxidase (PPO) method, myeloperoxidase (MPO) was also demonstrated in granulocytes. Peroxidase activity was exhibited by blast cells of acute myeloid leukaemia, including megakaryoblasts of megakaryoblastic leukaemia, by hairy cells of hairy cell leukaemia, by centroblasts of centroblastic lymphoma and by plasma cells of plasma cell leukaemia. The enzyme activity was also demonstrated in the mast cells of systemic mastocytosis. It seems that different forms of peroxidase are present in most haemic cells. Fixation and conditions of incubation are probably the determining factors for their demonstration.The method may allow further cytochemical discrimination at the LM level between blast cell populations which are negative for MPO by standard methods.

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Section: Methodsmentioning

confidence: 99%

“…As diffusion artefacts and non-enzymatic reactions with 3,3rdiaminobenzidine (DAB) can occur, the current method was accompanied by a set of controls. These included heat-treated films (15), cyanide-containing preincubation and incubation media (16) and films fixed and incubated according to the ordinary MPO technique (14,17).…”

Section: Methodsmentioning

confidence: 99%

Cytochemical demonstration of ‘platelet’ peroxidase at the light microscope level

Khalaf¹,

Hayhoe²

1987

European J of Haematology

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Neutrophil Myeloperoxidase Destruction by Ultraviolet Irradiation

Hanker¹,

Giammara²,

Strauss³

1988

Proc. annu. meet. Electron Microsc. Soc. Am.

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Only a fraction of the UV radiation emitted by the sun reaches the earth; most of the UVB (290-320nm) is eliminated by stratospheric ozone. There is increasing concern, however, that man-made chemicals are damaging this ozone layer. Although the effects of UV on DNA or as a carcinogen are widely known, preleukemia and acute myeloid leukemia (AML) have only rarely been reported in psoriasis patients treated with 8-methoxypsoralen and UV (PUVA). It was therefore of interest to study the effects of UV on the myeloperoxidase (MP) activity of human neutrophils. The peroxidase activity of enriched leukocyte preparations on coverslips was shown cytochemically with a diaminobenzidine medium and cupric nitrate intensification.Control samples (Figs. 1,4,5) of human bloods that were not specifically exposed to UV radiation or light except during routine handling were compared with samples which had been exposed in one of several different ways. One preparation (Fig. 2) was from a psoriasis patient who had received whole-body UVB phototherapy repeatedly.

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Neutrophil pseudoplatelets in subgingival plaque and crevicular fluid samples from periodontal diseased sites

Hanker¹,

Burkes²,

Greco³

et al. 1989

Proc. annu. meet. Electron Microsc. Soc. Am.

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The mature neutrophil with a segmented nucleus (usually having 3 or 4 lobes) is generally considered to be the end-stage cell of the neutrophil series. It is usually found as such in the bone marrow and peripheral blood where it normally is the most abundant leukocyte. Neutrophils, however, must frequently leave the peripheral blood and migrate into areas of infection to combat microorganisms. It is in such areas that neutrophils were first observed to fragment to form platelet-size particles some of which have a nuclear lobe. These neutrophil pseudoplatelets (NPP) can readily be distinguished from true platelets because they stain for neutrophil myeloperoxidase. True platelets are not positive in this staining reaction because their peroxidase Is inhibited by glutaraldehyde. Neutrophil pseudoplatelets, as well as neutrophils budding to form NPP, could frequently be observed in peripheral blood or bone marrow samples of leukemia patients. They are much more prominent, however, in smears of inflammatory exudates that contain gram-negative bacteria and in gingival crevicular fluid samples from periodontal disease sites. In some of these samples macrophages ingesting, or which contained, pseudoplatelets could be observed. The myeloperoxidase in the ingested pseudoplatelets was frequently active. Despite these earlier observations we did not expect to find many NPP in subgingival plaque smears from diseased sites. They were first seen by light microscopy (Figs. 1, 3-5) in smears on coverslips stained with the PATS reaction, a variation of the PAS reaction which deposits silver for light and electron microscopy. After drying replicate PATS-stained coverslips with hexamethyldisilazane, they were sputter coated with gold and then examined by the SEI and BEI modes of scanning electron microscopy (Fig. 2). Unstained replicate coverslips were fixed, and stained for the demonstration of myeloperoxidase in budding neutrophils and NPP. Neutrophils, activated macrophages and spirochetes as well as other gram-negative bacteria were also prominent in the PATS stained samples. In replicate subgingival plaque smears stained with our procedure for granulocyte peroxidases only neutrophils, budding neutrophils or NPP were readily observed (Fig. 6).

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Significance of Improved Cytochemical Methods for Hemoprotein Enzymes in Diagnosis and Classification of Leukemia

Cited by 3 publications

References 35 publications

Cytochemical demonstration of ‘platelet’ peroxidase at the light microscope level

Cytochemical demonstration of ‘platelet’ peroxidase at the light microscope level

Neutrophil Myeloperoxidase Destruction by Ultraviolet Irradiation

Neutrophil pseudoplatelets in subgingival plaque and crevicular fluid samples from periodontal diseased sites

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