Alpha-naphtyl Acetate Research Articles

Side Effects of In ovo Given Sunset Yellow FCF on the Embryonic Development of the Spleen by Means of Histological and Enzyme Histochemical Methods

Background: E110 is one of the food colorants which is widely used in dairy products, fast foods, jam and dry beverage powders, aqueous drug solutions, tablets, capsules, toothpastes mouthwashes, hair care products and cosmetics. Doubts have accumulated in recent years that food additives might cause allergic reactions in humans or increase these ailments. In this study, side effects of Sunset yellow FCF (E110) on the embryonic development of chicken spleen were evaluated by means of histological, histomorphometrical and enzyme histochemical methods. Methods: In the study, 250 fertilized broiler eggs obtained from a commercial broodstock were used. Eggs were divided into 5 groups each having 50 eggs. Control eggs were either nontreated or distilled water injected via air sac. The eggs in the experimental groups were injected with 100ng/egg, 500ng/egg and 1.000ng/egg E110 prior to incubation. Blood and spleen samples were taken from randomly selected 10 eggs of each group at 11th, 15th, 18th and 21st days of incubation. Leukocyte formula, alpha-naphtyl acetate esterase-positive and acid phosphatase-positive lymphocyte percentages were determined in the blood samples and embriyonic development of the spleen was assessed in the tissue sections. Results: In the 500ng/egg and 1.000ng/egg E110 injected experimental groups, embryonic development of the spleen retarded, alpha-naphtyl acetate esterase-positive and acid phosphatase-positive lymphocyte rates were significantly depressed. Conclusions: Significant disturbances in the immune system functions of the affected animals might occure at post-natal period of their life.

Effects of Aflatoxin on the Proportions of Peripheral Blood Leukocytes and Alpha-Naphtyl Acetate Esterase (ANAE) Positive Lymphocytes in the Mouse

Summary In this study detrimental effects of aflatoxin (AF) on the peripheral blood formula and the alpha naphthyl acetate esterase (ANAE) - positivity profile of peripheral blood lymphocytes (PBL) on the mice were determined. A total of 40 white mice were randomly divided in to 5 groups each having 8 animals. The first group served as a control and received standard diet. The remaining 4 groups were fed diets containing 200, 400, 800 and 1600 ppb AF, respectively. The results of this study have revealed that AF given by the food caused significant declines both in the proportion of PBL and in the percentages of ANAEpositive peripheral blood lymphocytes (T-lymphocytes) in a dose dependent manner. These results may be concerned as a haematological evidence for immuno-suppressive effect of AF on cell-mediated immunity.

Analysis of hydrolytic activity of phospholipase Cα from porcine retina on retinyl ester and phosphatidylcholine using non-denaturing two-dimensional electrophoresis and mass spectrometry

Hydrolysis of retinyl esters and phospholipids is important for visual functions of the animal retina. This study aimed to examine hydrolytic activity of an enzyme with native substrates such as retinyl esters and phospholipids responsible for this function in porcine retina. After cytosolic proteins were extracted from porcine retina, the proteins were separated using non-denaturing two-dimensional electrophoresis (2DE). Some major proteins and phospholipase Calpha were identified by matrix-assisted laser desorption ionisation-time of flight-mass spectrometry (MALDI-TOF-MS) or electrospray ionisation-tandem mass spectrometry (ESI-MS/MS). The phospholipase Calpha showed hydrolytic activities with not only alpha-naphtyl acetate but also with retinyl palmitate and phosphatidylcholine when effects of different substrates were investigated using enzyme activity staining on 2DE or MALDI-TOF-MS. Results indicated that hydrolytic activity of the enzyme with non-native and native substrates could be examined using a combination of non-denaturing 2DE and MALDI-TOF-MS.

Sialic acid 9-O-acetylesterase catalyzes the hydrolyzing reaction from alacepril to deacetylalacepril.

In this work, the alacepril thiolesterase, which catalyzes the hydrolyzing reaction of the thiolester linkage in alacepril and the conversion from alacepril to deacetylalacepril, was purified from rat liver cytosol and characterized. A purification procedure for the thiolesterase consisted of ammonium sulfate fractionation and chromatographies with phenyl Sepharose CL-4B, Q Sepharose FF, ceramic hydroxylapatite, and phenyl Sepharose HP. The thiolesterase activity was assayed for alacepril as a substrate and the reaction product, deacetylalacepril, was measured using high-performance liquid chromatography. The purified thiolesterase is heterodimeric with a molecular mass of 29 and 36 kDa subunits as estimated by sodium dodecyl sulfate -polyacrylamide gel electrophoresis. N-terminal amino acid sequence of these subunits reveals that the thiolesterase is identical to sialic acid 9-O-acetylesterase. The thiolesterase hydrolyzes not only the thiolester bond in alacepril, spironolactone, and acetyl coenzyme A but also the carboxylester bond in alpha-naphtyl acetate. The alacepril thiolestrase activity is competitively inhibited by alpha-naphtyl acetate. The thiolesterase, i.e., sialic acid 9-O-acetylesterase, seems to be involved in the metabolism of certain drugs such as alacepril and spironolactone. However, drugs having ester-type and amide-type linkages, for example dilazep. aniracetam, and benazepril, are not substrates for the thiolestrase.

Light and electron microscopic studies on alpha naphthyl acetate esterase activity of the peripheral blood T lymphocytes in van cats.

The purpose of this study was to determine the percentage of peripheral blood T lymphocytes and the localization of ANAE enzyme at the electron microscopic level in Van cats by using an alpha-naphtyl acetate esterase procedure. Peripheral blood samples taken from 20 Van cats were used. The percentage of ANAE positive lymphocytes was 83.0%. Neutrophilic granulocytes gave a negative reaction, whereas monocytes, eosinophilic granulocytes showed a diffuse granular positivity. In the electron microscopic examination, ANAE positive reactions were seen in lysosomal granules found in lymphocytes.

Selective action of androgens on the molecular forms of esterases characterized by two‐dimensional gel electrophoresis in the epididymis and vas deferens of the mouse

Molecular forms of esterases were resolved in non-denaturing conditions by using two-dimensional gel electrophoresis with isoelectric focusing in the first dimension and a time-dependent polyacrylamide gradient gel electrophoresis (PAGGE) in the second dimension. This procedure was used to analyse sequential changes in esterase composition along the excurrent genital duct of the mouse and to initiate a specific identification of the androgen-regulated molecular forms. Almost all the 68 variants (pH 3.9-6.4 and 50-300 kDa) revealed by alpha-naphtyl acetate from the fluids of the three parts of the epididymis (caput, corpus, cauda) and vas deferens, could be assigned to the carboxylesterase group as shown by their action on various substrates and sensitivity to inhibitors. Some of these variants co-migrated with those in the serum and testis, whereas other enzyme forms made their first appearance in the caput (13), in the corpus (26) and in the vas deferens (3). The major changes occurred between the caput and the corpus of the epididymis. Only a few acidic spots were not revealed after neuraminidase digestion. Castration of mice (4 weeks) resulted in inhibition of the activity of 34 esterase forms, and thus abolished most of the regional differences in the excurrent duct system. By re-initiating or repressing the synthesis of regional esterase variants, testosterone supplementation (2 and/or 4 weeks) of castrated animals restored the normal esterase pattern in the three epididymal parts, but not in the vas deferens. The major effect of efferent duct ligation (4 weeks) was the emergence in the corpus and cauda of the epididymis of two variants found in the caput of uncastrated mice.

The Langerhans' cells.

The morphological, enzymatical, immunocytochemical and functional properties of Langerhans' cells are briefly reviewed. Langerhans' cells are located mainly in the squamous stratified epithelia, but are also present in the thymus and in superficial lymphnodes. At the ultrastructural level, they are characterized by unique cytoplasmic organelles, the Birbeck granules, whose function is still unknown. Langerhans' cells possess strong ATPase activity and are weakly positive for alpha-naphtyl acetate esterase and for acid phosphatase; they are immunoreactive for CD1a (T6), class II MHC antigens and S-100 protein. In some pathological conditions, like dermatopathic lymphadenopathy and Langerhans' cell histocytosis, Langerhans' cells also are characterized by the expression of monocyte-macrophage antigens. Langerhans' cells act as antigen-presenting cells to T lymphocytes; their functional capacity is strictly dependent on the levels of expression of class II MHC antigens. Langerhans' cells are of bone marrow origin and are derived from a circulating precursor which is probably related to the monocyte.

Characterization of factor XIII containing-macrophages in lymph nodes with Hodgkin's disease.

A large number of cells containing subunit a of blood coagulation Factor XIII (FXIII) was detected by immunoperoxidase staining in lymph nodes with Hodgkin's disease. These relatively large, multipolar, mononuclear cells were often found in the immediate vicinity of malignant Hodgkin's cells. Intensive characterization of these cells carried out by immunofluorescent and enzymecytochemical techniques in double- and triple-labelling systems on the same sections clearly demonstrated that they represent tumour-associated macrophages (TAMs). FXIII containing-cells showed alpha-naphtyl acetate esterase (ANAE) positivity, and were labelled by monoclonal anti-Leu M3 antibody, a monocyte/macrophage marker, but not at all or only very weakly by anti-HLA-DR. Neither alkaline phosphatase (ALP) nor adenosine triphosphatase (ATPase) activity could be detected in these cells and surprisingly, they were consistently negative for acid phosphatase (AcP) as well. The presence of FXIII subunit a in tumour-associated macrophages suggests that this cell type might have an important role in the stabilization of fibrin deposits around tumour cells.

Leukemia of early infancy: Early B-cell lineage associated with t(4:11)

A case of infantile acute leukemia associated with translocation t(4:11)(q21:q23) is reported. This leukemia has a very poor prognosis, and this patient survived for only 9 months. The blast cell morphology was L1/L2 according to the FAB classification and showed a lymphoid appearance on transmission electron microscopy. The histochemical stains showed a pattern of periodic acid-Schiff positivity and variable alpha-naphtyl acetate staining. The cells were TdT-positive and surface-marker phenotyping was positive for Ia-like and B4 antigens but negative for CALLA, T-cell markers, myelocyte and monocyte markers. The leukemic cells represent a frozen state of a very early precursor, corresponding to the earliest recognizable stage of the B-cell lineage. This observation may contribute to the controversion regarding the cell origin of this unique leukemia associated with t(4:11), lymphatic versus null cell, early myeloid, or mixed, and points to the possibility of a very early B-cell lineage leukemia.

Lymphoid cells in the venous blood of 100 female patients with breast carcinoma. Follow-up studies

The following blood cells were examined in 100 patients with invasive carcinoma of the breast and 100 control patients without malignant disease: leukocytes, lymphocytes, monocytes and different sub-populations of lymphoid cells. Lymphoid cells were characterized by E-, EA- and EAC-rosetting technique, alphanaphtyl acetate esterase activity and by the ability of latex phagocytosis. Both groups showed no significant differences. In 81 patients undergoing postoperative radiation therapy lymphocytes and lymphoid cells were reduced considerably. These changes were still evident after 12 months (6 months after completion of radiotherapy). Mainly the T lymphocytes were effected by this decrease. A negative effect of the radiation induced reduction of immune competent cells must be discussed and examined further by clinical trials.

Complement receptor distinguishes between two subsets of large granular lymphocytes with different natural killer activity and cytochemical and ultrastructural features.

Human peripheral blood large granular lymphocytes (LGL)--that is, cells with intracytoplasmic azurophilic (electron-dense) granules, with a positivity for the cytochemical localization of certain acid hydrolases, and with avid surface receptors for the Fc portion of IgG--have been purified on Percoll density gradients. Approximately 30% of these cells expressed receptors for the third complement component (C3R). They were separated into C3R-positive and C3R-negative cells. C3R+ cells had a significantly greater natural killer (NK) activity against K562 target cells than C3R+ cells. This difference was unrelated to the presence in the C3R+ cells of a contaminant cell type incapable of NK activity, since cytochemical and ultrastructural analysis revealed that C3R+ and C3R- fractions contained comparable LGL numbers. Agarose cytotoxicity assays at the single-cell level demonstrated that C3R+ LGL contained a large number of cells that bound to but did not lyse the target. The remaining fully cytotoxic C3R+ LGL had, however, the same killing and recycling properties as the cells from the C3R fraction. Electron microscopy and cytochemical studies showed that C3R+ cells had fewer electron-dense granules than C3R cells and stained more faintly for the localization of alpha-naphtyl acetate esterase. In contrast to C3R cells, C3R+ LGL displayed morphological features suggesting that an active process of granule formation was taking place. Taken together, the data indicate that C3R+ cells represent a discrete subset or a maturational stage of LGL.

Demonstration of T Lymphocytes in Leprous Granuloma Using the Acid α Naphtyl Acetate Esterase Activity: An Attempt at Quantitative Analysis

In nine leprosy patients (1 TT, 1 BT, 4 BL and 3 LLp), esterase positive lymphocytes (T Lymphocytes) were studied in frozen sections of skin biopsies by alpha naphtyl acetate esterase pH 5.8 method (ANAE) Four patients had never been treated previously and five patients exhibited clinical and bacteriological evidence of relapse for inadequate therapy at first biopsy. There was an increase in ANAE (+) lymphocyte density in granulomas when second biopsies were done after efficient treatment, evaluated by bacillary index for the eight bacilliferous patients, and clinical improvement. The significance of T cells in granulomas is discussed.

Dendritic reticulum cell sarcoma? Four cases of a lymphoma probably derived from dendritic reticulum cells of the follicular compartment

This paper describes the histological picture of four tumours of the follicular compartment of the lymph node, in which the proliferating cell appeared to be the dendritic reticulum cell (DRC). This assumption was based on the results of light microscopical, ultrastructural, immunological, and enzymehistochemical investigations. The tumour cells resembled DRC's closely in (1) the striking pattern of interdigitations and occasional tight junction-like contacts between the neoplastic cells on electron microscopical analysis; (2) presence of receptors for the activated third component of complement on the membrane of the cells; (3) absence of monoclonal immunoglobulins and T-cell antigen on the surface and of lysozyme, alpha 1-antitrypsin or alpha 1-antichymotrypsin in the cytoplasm of the neoplastic cells. Moreover, (4) the tumour cells showed moderate alpha-naphtyl acetate esterase, weak to absent acid phosphatase and (with one exception) strong 5-nucleotidase activity. Furthermore, (5) the neoplastic cells expressed Ia-like antigens on the surface in all four cases. The relation with follicle centre cell lymphomas, the differential diagnosis and clinical data are discussed.

Cytochemical profile of human haematopoietic biopsy cells and derived cell lines.

Twenty-three human haematopoietic cell lines, normal and mitogen stimulated peripheral blood lymphocytes and tumour material from fresh leukaemias, myelomas and lymphomas were investigated with a panel of cytochemical reactions. Normal and mitogen stimulated lymphocytes, non-neoplastic lymphoblastoid cell lines (LCL), lymphoma lines with B-lymphocyte characteristics, chronic lymphocytic leukaemia and fresh lymphocytic lymphomas reacted weakly or negatively with all stains. T-lymphocyte acute leukaemia lines were PAS and alpha-naphtyl acetate esterase positive. Myeloma lines and fresh myelomas were strongly beta-glucoronidase positive. A histiocytic lymphoma cell line was strongly esterase positive with naphtol AS-D acetate esterase inhibited by NaF. The three fresh histiocytic lymphomas, however, reacted as the lymphocytic lymphomas suggesting a lymphoid origin. A myeloid leukaemia line was strongly positive for acid phsophatase. No major disagreement was noted between the reactivity of established neoplastic lines and the corresponding fresh biopsy cells indicating an unaltered qualitative expression of enzyme production after prolonged in vitro culture.