Naphthol AS-D Acetate Esterase Research Articles

The proliferation of cells in the dermal infiltrate of lichen planus lesions.

The proliferation of mononuclcar cells infiltrating the dermis in lichen planus lesions was studied in five untreated patients. A biopsy of a lesion was made in each patient. Thin slices of the specimens were incubated for 60 min in the presence of 3H thymidine (3HT), fixed and radioautographed. The mean value of the 3HT labelling index (±SE) was 2.87±0.14%. This labelling index (LI) is relatively high; it is not statistically different from the LI encountered in the dermal infiltrate of allergic contact dermatitis (P>0.45), but is three to four times higher than in psoriasis or irritant dermatitis. The proliferative cells are most probably lymphoblasts. A biopsy of another lichen planus lesion was made in two of the five patients 60 min after a local injection of 10 μCi of 3HT. Imprints of lesions were treated histochemically for Naphthol AS-D acetate esterase(s) identification, a useful method for distinguishing monocytes (many cytoplasmic granules) and lymphocytes (few if any granules). The imprints were then radioautographed. No 3HT-labelled cells contained more than one or two cytoplasmic granules, which suggests that they are of lymphocytic origin. The biological significance of this lymphoblastic proliferation still remains hypothetical.

Splenic hemangiosarcoma with sinus endothelial differentiation

A case of primary splenic hemangiosarcoma where the tumors entirely replaced the spleen is reported. Histologically, the splenic tumors consisted of two major components: (i) solid and nodular areas, in which spindle cells (SC) were arranged in interlacing fascicles; and (ii) vasoformative areas, in which vascular lining cells (VC) closely resembled normal splenic sinus endothelial cells (SSEC) in shape. Both SC and VC were immunoreactive with the markers of vascular endothelial cells (EC) including SSEC, JC70, von Willebrand factor, OKM5, vimentin MAb and Ulex europaeus-1 lectin. The tumor cells were positive for OKM5 MAb as well as normal SSEC and sheathed arterial EC, but other vascular EC in normal spleen were negative. Enzyme-histochemically, the tumor cells and normal SSEC revealed naphthol AS-D acetate esterase activity but no reaction for ALPase, whereas normal vascular EC other than SSEC in the spleen were positive for the latter but negative for the former. These studies revealed that the tumor cells expressed the same phenotype as normal SSEC. To the present author's knowledge this is the first report demonstrating sinus endothelial differentiation of splenic hemangiosarcoma, enzyme- and immuno-histochemically.

Histochemical Study of Non-Specific Esterases Motor Unit of the Rabbit

The distribution of alpha naphthyl asetrase and naphthol AS-D acetate esterase was studied in the motor neurons of the ventral horn of the spinal cord segments L4, L5, and SI, supplying the extensor digitorum longus musscle (EDL) of the rabbit together with the peripheral nerve supplying EDL and the muscle itself. 
 Strong activity was demonstrated in the motor neurons, in schwann cells and in the axons of peripheral nerve bundlds. Abundant reaction was located in the motor end-plates of muscle fibres making them easily recognizable. biological interpretation of esterases was discussed

Acute Promyelocytic Leukaemia. Relationship of Cytochemical Pattern with Clinical, Morphologic and Immunological Characteristics, and Outcome.

In a series of 61 acute promyelocytic leukemia (APL) cases multiple clinical and biological characteristics were studied in order to assess their prognostic significance and to evaluate the relationship of cytochemical pattern with clinical, morphologic, and immunological characteristics and outcome. Alpha-naphthyl acetate esterase (ANAE) activi1:y was expressed in 23 out of 41 cases (56%). Sodium fluoride-sensitive naphthol AS-D acetate esterase was evident in only one out of 22 cases (5 %). ANAE reactivity was more frequently found in typical hypergranular APL (P = 0.003) and was associated with a shorter interval symptoms-diagnosis (P = 0.04). The expression of HLA-DK, CD33, CD13 and CD11b did not show any correlation with ANAE reactivity. Complete remission (CR) was achieved in 64 % of treated patients. The actuarial median remission duration was 22 ± 3 months. CR rate was significantly related to the presence of fever, age, absolute reticulocyte count, and albumin level, while the remission duration was associated with absolute reticulocyte, leucocyte and granulocyte counts, and albumin level. In our study ANAE activity did not reflect any important clinical and biological features of APL, apart from its obvious correlation with morphological subtype. In addition ANAE activity did not correlate with early response to therapy nor related with long-term prognosis.

Nonspecific esterase of acute promyelocytic leukemia (M3).

Leukemic cells of 43 patients with acute promyelocytic leukemia (M3) were investigated morphologically and cytochemically to determine the percentage of aberrant enzymes and whether or not the presence impacts on the clinical outcome. Twelve patients (27.9%) showed alpha-naphthyl acetate esterase (ANAE) activity in their leukemic cells, and two of these cases revealed remarkably low myeloperoxidase (MPO) positivity, a pattern seen in monocytic precursors. However, further cytochemical evidence for this monocytic feature, the inhibition of naphthol AS-D acetate esterase (NASDA) activity with sodium fluoride (NASDA-F), was found in only five of these nine patients. In 31 cases (72.1%), there was minimal ANAE activity. Of interest, two of these were devoid of naphthol AS-D chloroacetate esterase (CAE), which is prominently displayed in neutrophilic granulocytes, even though these leukemic cells were 100% intensely positive for MPO activity. Between the two groups with and without ANAE activity, there were no remarkable differences in the distribution of sex and age, hematological findings, and rates of complete response. Our study has confirmed the cytochemical heterogeneity of M3 with no obvious relationship between this heterogeneity and early therapeutic outcome.

Isolation of osteoclasts from pagetic bone tissue morphometry and cytochemistry on isolated cells

Giant osteoclasts and other cells were isolated from Pagetic bone tissue using 0.5 mM ethylene diamine tetraacetic acid on bone samples from 8 patients with Paget's disease. The cell suspension contained osteoclasts and osteoblasts as well as some mononuclear cells such as monocytes. The number of nuclei in isolated osteoclasts (33.85 ± 20.92 nuclei/osteoclast) correlates fairly well ( p < 0.02) with the number of nuclei counted on histologic sections (15.88 ± 11.80 nuclei/osteoclast) for samples from each patient. Enzyme histochemistry demonstrated acid phosphatase activity in isolated osteoclasts and in mononucleated cells, such as monocytes. Alkaline phosphatase was detected only in osteoblasts while succinate dehydrogenase was observed in osteoclasts, osteoblasts and monocytes. Esterases, such as nonspecific aliesterase and specific naphthol AS-D acetate esterase, were identified in osteoclasts and in macrophages. Inhibition of specific naphthol AS-D acetate esterase in osteoclasts by addition of sodium fluoride suggests that the enzyme could be of monocytic origin.

Distribution of hydrolytic enzymes at barley powdery mildew encounter sites: implications for resistance associated with papilla formation in a compatible system

Distributions of the hydrolytic enzymes acid phosphatase and non-specific esterase in compatible barley coleoptiles inoculated with Erysiphe graminis fsp. hordei were investigated histochemically by light and electron microscopy. The four different enzymes investigated-acid β-glycerophos-phatase, acid naphthol AS-TR phosphatase, indoxyl acetate esterase and naphthol AS-D acetate esterase—had similar localization patterns. Reaction products were distributed generally in the fungal and host cells, but distinct accumulations were found in certain regions and structures at the encounter sites, including appressoria, penetration pegs, haustoria, papillae, and host cytoplasm. Most papillae showed conspicuous amounts of reaction products, suggesting that papilla formation includes depositions from secretory vesicles derived from the lysosomal system in host cells. There was a marked difference in the apparent concentration of hydrolytic enzyme activity between normal-size papillae accompanied by haustoria (sites of successful penetration) and oversize papillae (sites of unsuccessful penetration). Hydrolytic enzymes, localized most intensively in a lAyer of the oversize papillae, may confer resistance to fungal ingress by degrading components of the penetration pegs.

Pure Acute Monocytic Leukemia: A Study of 12 Cases

Twelve cases of pure acute monocytic leukemia in adults were studied. They were selected on the basis of the morphology of the blast cells on Romanowsky-stained smears of blood and bone marrow, as well as positivity of the cells for the naphthol ASD acetate esterase reaction specifically inhibited by sodium fluoride. There was no sex predominance. Neoplastic involvement of the skin and/or gingiva was very frequent. The leukemic proliferation in blood and bone marrow consisted of monoblasts, promonocytes and monocytes. The peroxidase reaction was negative or only faintly positive. Serum and urinary lysozyme levels were increased. The blast cells retained their ability to stimulate, in vitro, colony formation by normal bone marrow cells used as targets. All of these characteristics permit specific identification of this type of acute leukemia. The prognosis is grim: only five of 12 patients achieved complete remission, and four of these five had relapses in less than 14 months; the median survival was five months.

Nonoxidative fungicidal mechanisms of mammalian granulocytes: demonstration of components with candidacidal activity in human, rabbit, and guinea pig leukocytes.

Granulocytes from the peripheral blood of normal subjects and a patient with hereditary myeloperoxidase deficiency were homogenized in 0.34 M sucrose. A granule-rich fraction, prepared by sedimentation at 27,000 x g for 20 min, contained components that killed C. parapsilosis in vitro. These were extractable with 0.01 M citric acid and were shown by micropreparative polyacrylamide electrophoresis to be multiple. The candidacidal activity of these neutrophil components was heat stable and they were somewhat more active at pH 5.0 than at pH 7.0. When rabbit or guinea pig heterophils were obtained from sterile peritoneal exudates and similarly fractionated, they also were found to contain components that killed C. parapsilosis in vitro. These were primarily associated with a group of lysosomal cationic proteins lacking direct counterpart in human neutrophils. Among the candidacidal components of the human neutrophil was a protein, more cationic than lysozyme, that exhibited naphthol-ASD acetate esterase activity.

“Pure” monocytic or histiomonocytic leukemia: A revised concept

In a series of 130 cases of acute leukemia studied by cytochemical staining techniques, 10 cases cytochemically diagnosed as "pure" monocytic leukemia were seen. Cytochemical staining of bone marrow aspirates from these patients revealed all leukemic cells to be Sudan black negative. No positive reactions were observed for peroxidase or naphthol AS-D chloroacetate esterase. All cases demonstrated strong alpha-naphthyl acetate esterase positivity; and fluoride-inhibited naphthol AS-D acetate esterase positivity was observed in 8 of 9 cases tested. The P.A.S. reaction showed diffuse fine to coarse granules. Oil red O stain was positive in 8 of 9 cases, and the beta-glucuronidase activity was strong in 5 of 9 cases. Light microscopy revealed cells with monocytic or histiocytic morphology. Electron microscopic studies in 2 cases demonstrated features consistent with leukemic monocytic or histiocytic morphology; none was suggestive of granulocytic or lymphocytic leukemia. Five of 6 patients treated with drug regimens including prednisone and vincristine entered a complete remission; the other obtained a partial remission. Two patients achieved complete remission after treatment with Adriamycin, 1 following a relapse. Three patients who received cytosine arabinoside as their only therapy died soon after treatment was commenced. It is suggested that the cytochemical similarity but morphological differences in those patients may be objectively used to group them as cases of histiomonocytic leukemia.

Soluble proteins from human leukocyte granules. I. Esterase activity of cationic proteins.

Soluble granule proteins from human granulocytes were subjected to acrylamide-gel electrophoresis at pH 4.3. The gels subsequently were reacted for peroxidase and 11 hydrolytic enzymes (i.e. acid phosphatase, alkaline phosphatase, alanine aminopeptidase, arylsulphatase, β-galactosidase, β-glucuronidase, leucine aminopeptidase, α-naphthylacetate esterase, naphthol AS-D acetate esterase, naphthol AS-D chloroacetate esterase and lysozyme). Out of these activities only esterase activity (12 bands) on the 3 ester substrates and lysozyme activity (1 band) could be identified with the proteins showing high cathodal migration. 2 of the proteins showing esterase activity migrated faster to the cathode than the lysozyme. Esterase activity was sensitive to di-isopropylfluorophosphate.