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The aim of this study is to determine the functional effects of the acrylamide (AA) administrated by oral gavage on the peripheral blood lymphocytes (PBL) and the Gut Associated Lymphoid Tissue (GALT) in male Sprague-Dawley rats using alpha-naphthyl acetate esterase (ANAE) demonstration. For this purpose, two separate experiments were performed with Sprague Dawley rats. In Experiment-I rats were gavaged with 0, 30, 45 and 60 mg/kgb.w. AA for five consecutive days and in Experiment-II rats were gavaged with 0, 125, 150, and 175 mg/kg/b.w. AA for single oral dose. Animals were sacrificed 24 h after the last treatments in both experiments by servical dislocations under ether anaesthesia. Blood samples were collected from the heart in heparinized (10 UI heparin/ml(-1) of the blood) tubes before sacrification and lymphoid tissue samples from the ileal Peyer's patches (IPPs) were taken and processed for histochemical demonstration of ANAE following the sacrification. The lymphoid follicles of the IPPs of animals given 125, 150 and 175 mg/kgb.w. AA were markedly reduced in size. Germinal centres (GCs) markedly regressed in AA-treated animals compared with those of controls. ANAE-positive lymphocyte depletion of IPPs was very prominent in the high doses AA-treated animals. In the animals treated with 30, 45, and 60 mg/kg b.w. AA, the IPPs had similar histology to those of the controls. ANAE-positive peripheral blood lymphocyte levels significantly decreased in AA exposed groups in a dose dependent manner (p

Plant-esterase from wheat flour was purified via aqueous two-phase system (ATPS). The enzyme crude extract was first filtered through a polyethersulfone OmegaTM ultrafiltration membrane, followed by two-step ATPS with PEG1000, NaH2PO4 and (NH4)2SO4. Based on previous studies, the first ATPS consisting of 27.0% PEG1000 and 13.0% NaH2PO4 (w/w) was performed at room temperature and pH 5.0. The top phase was carefully separated, and 6.0% (NH4)2SO4 (w/w) was added to form the second phase system under similar conditions, to obtain the pure enzyme that partitioned in the bottom salt phase. The purification thus obtained was 10.35 fold with an enzyme yield of 72.90%. Sodium dodecylsulphate polyacrymamide gel electrophoresis (SDS-PAGE) was used to estimate the molecular weight of the target esterase to be around 68 kilo Daltons (kDa). Using a 16 mM solution of alpha naphthyl acetate as substrate, the maximum activity of the purified enzyme, alpha naphthyl acetate esterase (ANAE) was found to be 0.78 U at the optimum conditions of 40 ◦C and pH 8.0 for 15 min of incubation. The Michaelis–Menten parameters Km and Vmax of the purified enzyme were estimated using Langmuir linear plot as 22.5 mM and 4.71 U mg−1, respectively. Further purification was obtained by dialyzing the bottom ANAE rich phase in a solution of 0.04 M phosphate buffer with pH 6.5 to remove the salt, which was then concentrated using centrifugal tubes with OmegaTM membrane. The highly pure ANAE so obtained was immobilized on the multiwalled carbon nanotube for the development of inhibition based enzyme biosensors for pesticides detection.

The average percentage of acid alpha naphthyl acetate esterase reacting lymphocytes (APARL) was enumerated in the peripheral blood of chickens challenged with Marek's disease after vaccination with either turkey herpesvirus (HVT), inactivated Marek's disease virus (IMDV) or a mixture of the two (bivalent vaccine). A gradual increase in APARL value was noticed in the vaccinated chickens from day 7 to 70 after challenge with a virulent Marek's disease virus. The increase was consistent and significantly higher in bivalent (HVT plus IMDV) than in HVT-vaccinated chickens while the slight increase noticed in IMDV vaccinated-challenged birds was inconsistent.

The histochemical demonstration of nonspecific acid α-naphthyl acetate esterase (ANAE) activity was evaluated as a T lymphocyte marker primarily with the sheep erythrocyte (E) assay. A distinctive staining pattern characterized T lymphocytes which could be readily distinguished from monocyte staining. The percentage of E + and ANAE + lymphocytes was nearly always comparable in the peripheral blood and lymphoid tissue from normal and selected patients, including those with acute and chronic lymphocytic leukemia. Divergences were noted in certain other tissues including spleen and thymus. Certain mitogen-stimulated cells lost their ANAE activity while retaining their ability to form e n rosettes. Atypical and variable staining patterns were observed in established lymphoid cell lines. The histochemical demonstration of ANAE is simple and reproducible; preparations may be counterstained for cytomorphologic detail and mounted as a permanent record. Certain disadvantages are discussed. The method represents a practical alternative to E rosette assays. It is particularly well suited for certain routine laboratories.

Nonspecific acid alpha naphthyl acetate esterase (ANAE) activity has been demonstrated in bovine lymphocytes and monocytes-macrophages. The lymphocytes exhibited a solitary reddish brown globular reaction adjacent to the cell membrane (T-pattern) whereas the monocytes-macrophages revealed a diffuse reddish brown cytoplasmic reaction (M-pattern) product. Cy tocen tri fuge smears of lymphocytes forming rosettes with sheep erythrocytes also revealed T-pattern ANAE reaction in the majority of the cells. Similar T- and M-patterns of ANAE activity were observed in cryostat sections of biopsies collected from normal bovine lymph nodes and horn cancer tissue sections from bullocks.

A strong activity of acid alpha-naphthyl acetate esterase (ANAE) in T lymphocytes and macrophages within mouse lymphoid organs is demonstrable even after paraffin-embedding, if a rapid embedding procedure with acetone as dehydrating agent is used. The lymphoid tissue structure is better preserved in paraffin sections procured after this rapid embedding procedure than in frozen sections commonly used for histochemical demonstration of ANAE. In the paraffin sections the pattern of the ANAE reaction allows a clear delineation of the T and B areas in the lymphoid organs, and the intracellular location of the focal ANAE activity in T lymphocytes is readily discernible.

Alpha-naphthyl acetate esterase (ANAE) activity was demonstrated in ovine lymphocytes harvested from blood on Ficoll-metrizoate gradients. The enzyme's specificity for T lymphocytes, identified by immunofluorescent staining of T cell-specific antigens, was assessed. Correlation analysis of the results obtained using unfractionated lymphocytes from 12 sheep showed no correlation between ANAE activity and the expression of T cell antigens ( r = 0.22). When lymphocytes from 4 sheep were fractionated on nylon wool columns a mean of only 43.2% of the cells in the non-adherent population were ANAE-positive whereas 94.7% of these cells were identified as T lymphocytes. Blood lymphocytes from 5 animals were separated into 3 fractions using Percoll discontinuous density gradients. No significant relationship was seen between ANAE activity and T cells in Fractions 1 and 3 ( r = 0.41 and 0.21). Fraction 2 cells, however, did show a significant positive relationship ( r = 0.91) between these two features but the biological significance of this relationship is unknown. It was concluded that ANAE activity is not a specific marker for ovine T lymphocytes.

The distribution of mononuclear cell subsets has been studied in human adenoids, tonsils and peripheral blood (PB) by evaluating the presence of surface immunoglobulins, E-rosette formation, receptors for IgG Fc and for complement, alpha-naphthyl acetate esterase (ANAE) cytochemistry, reactivity with peanut lectin (PNA) and with monoclonal antibodies (McAb) (OK panel). Adenoids and tonsils, compared to PB, contain (1) fewer macrophages and T cells but more B cells; (2) higher proportions of ANAE negative, complement receptors and Ia-like antigens bearing T lymphocytes; (3) higher percentages of cells reacting with the McAbs OKT9 and OKT10 ("immature" lymphoid cells). In both adenoids and tonsils, clusters, formed by a central heavily ANAE stained interdigitating cell surrounded by lymphocytes with a sickle-shaped ANAE reaction, were found. Analogous clusters have been previously described in mice and human thymus. Two major hypotheses could be put forward: (1) adenoids and tonsils contain "immature" lymphoid cells undergoing education process, or (2) the above organs contain lymphocytes activated by a constant exposure to bacterial antigens or mitogens.

In acute myeloid leukemia the leukemic cells are thought to be blocked in their normal differentiation. The stage of differentiation is the basis for the FAB classification. Induction of cell differentiation is a new and promising development in the treatment of some myeloproliferative diseases. The criteria for cell maturation and differentiation are almost all based on the morphology of the leukemic cells. In this study we tried to specify the maturation stage of the leukemic cells by quantitative enzyme analysis. In the HL60 (promyelocytic) cell line cells we determined the following enzymes: myeloperoxidase; alpha naphthyl acetate esterase; alpha naphthyl butyrate esterase and lactate dehydrogenase. The enzyme profiles obtained after culturing the cells in the presence of the differentiation inducing agents 1 : 25 dihydroxy vitamin D3 and DMSO were compared with several cytochemical and functional parameters. The results obtained in this study show that quantitative enzyme analysis is a useful tool in the study of myeloid or monocytic differentiation.

Human foetal mononuclear cells from thymus, spleen, liver, bone marrow and peripheral blood at 8–24 weeks of gestation were examined for cytochemical evidence of acid alpha-naphthyl acetate esterase (ANAE) activity. The focal brownish-red ANAE reaction product (T cell staining pattern) was observed in counterstained cytocentrifuged cell smears in the cytoplasm. ANAE-positive lymphoid cells were first observed in the thymus at 9 weeks of gestation. A gradual increase in frequency of ANAE-positive cells in foetal thymus was observed, from about 10% at 14–15 weeks to about 20% at 22–24 weeks of gestation. By 14 weeks of foetal age, spleen and liver contained a few ANAE-positive cells and after 15 weeks of gestation consistent occurrence of ANAE-positive cells was observed in foetal bone marrow and peripheral blood. These results demonstrate that ANAE-positive lymphocytes first appear in the foetal thymus and are subsequently found in the foetal liver, spleen, bone marrow and peripheral blood.

The T lymphocytes (alpha naphthyl acetate esterase positive cells) were estimated in the peripheral blood smears as well as in the separated lymphocytes in tumour bearing and non-tumour bearing control cattle. The results obtained with both techniques were similar and it was suggested that the blood smear staining technique could be routinely used in place of the time consuming and laborious cell separation technique. The T lymphocyte percentage in the peripheral blood of the tumour bearing animals was lower than that of healthy control animals. However, the animals in stages 1 and 2 showed slightly higher values than the stage 3 tumour bearing cattle.