Α-naphthyl Acetate Esterase Research Articles

The role of juvenile hormone and brain factor(s) in the regulation of plasma juvenile hormone esterase activity during the last larval stadium of the tobacco hornworm, Manduca sexta

During the last (fifth) larval stadium of gate-1 Manduca sexta there were two peaks of juvenile hormone esterase activity in the plasma. The first peak occurred on day 3 preceding wandering and the second of a similar size on day 8 preceding pupation. The topical application of juvenoids (juvenile hormone I, epofenonane, and ( RS)-methoprene) to the abdomens of prewandering (day 0 to 3) and postwandering (day 5 and 7) larvae increased the plasma juvenile hormone esterase activity. Head ligation and starvation of prewandering larvae blocked the first peak and ligation of postwandering larvae blocked the second peak. Application of juvenoids could not rescue the effects of ligation and starvation in prewandering day 0 to 2 larvae, but ligation of day 1 and 2 larvae for 24 h followed by juvenoid treatment restored normal juvenile hormone esterase activity levels in ligated but not starved larvae. Juvenile hormone acted directly on the abdomen presumably on the fat body to increase juvenile hormone esterase activity throughout the last stadium but day-3 and -7 larvae were most responsive. Injection of head extracts of day-1, -2 and -3 fifth-instar larvae into the abdomens of head-ligated day-1 larvae did not increase the plasma juvenile hormone esterase activity, whereas it increased the α-naphthyl acetate esterase activity. Day-1 head extract decreased the plasma juvenile hormone esterase activity in unligated day-0, -1 and -6 larvae and in 24 h head-ligated, day-2 larvae that were treated with epofenonane. In day-1 unligated larvae treated with epofenonane one head equivalent reduced the juvenile hormone esterase activity by 48%. Injection of 4 head equivalents from day-1 larvae into larvae of the same age not treated with juvenoids completely inhibited the normal increase in juvenile hormone esterase activity seen between day 1 and 2. The inhibitory factor was specific in action not affecting the α-naphthyl acetate esterase activity, was dose-dependent, and found both in head and brain but not in whole body extracts. The inhibitory effect was present in head extracts prepared from day-1, -2 and -3 larvae. Head extracts also significantly decreased the juvenile hormone esterase activity in the whole body of normal as well as epofenonane-treated larvae without affecting the α-naphthyl acetate esterase activity.

Acid .ALPHA.-naphthyl acetate esterase(ANAE) staining of porcine lymphocytes.

Acid α-naphthyl acetate esterase (ANAE) activity was investigated to be one of porcine T lymphocyte markers by modified fixation procedures in normal porcine lymphocytes prepared from spleen, blood, mesenteric lymph nodes, thymus and Peyer's patch. Smears were fixed for 2 to 3 seconds with buffered formalin acetate (pH 6.6). The majority of ANAE positive cells had the red-brown spot. The cells diffusely stained with densely red color were found to be monocytes. Other stained cells had stained granules. There were no differences between the ratios of ANAE positive cells with stained spots or granules and those of E-rosette positive cells in lymphocyte preparations from spleen, blood, mesenteric lymph nodes and Peyer's patch. However, the ANAE positive cells in thymus were stained faintly and the ratio of ANAE positive cells was by far lower than that of E-rosette positive cells. In Con A or PHA-P stimulated lymphocytes, ANAE staining density was more faint than that of non-stimulated cells and the percentages of ANAE positive cells were lower than those of E-rosette positive cells. These results indicated that ANAE activity could be one of T cell markers for porcine mature lymphocytes (spot or granular pattern) of spleen, blood, mesenteric lymph nodes and Peyer's Patch and one of markers for porcine monocytes (diffuse pattern).

Relationship between acupuncture-induced immunity and the regulation of central neurotransmitters in the rabbit: I. Effect of central catecholaminergic neurons in regulating acupuncture-induced immune function.

The central catecholaminergic neurons play an important role in the promotion of the T-lymphocyte transformation and the regulation of the positive rate of acid a-naphthylacetate esterase (ANAE) activity with acupuncture. The mechanism by which acupuncture promotes the T-lymphocyte transformation function may be principally by promoting the anabolism of the T-lymphocyte and speeding up the anabolism of the DNA. Judging from the fact that acupuncture increases the positive rate of ANAE, it can be assumed that the quantity of T-lymphocyte was increased markedly. It remains to be clarified what links exist through which the central catecholaminergic neurons can influence the acupuncture induced immunity, and what the effect of other transmitter neurons of the central nervous system are on acupuncture induced immunity.

Acid α-naphthyl acetate esterase assay in murine lymphocytes

Acid α-naphthyl acetate esterase (ANAE) activity was assayed in cell homogenates and in intact cells by an endpoint colorimetric method, in which sodium dodecyl sulfate was used to stop the reaction. Each method of cell disruption and enzyme solubilization tested here caused a partial loss of the ANAE activity in lymphocyte preparations. The majority of the ANAE activity in lymphocytes was found to be membrane bound. The ANAE activity in thymocytes was over two times lower than that obtained for lymph node and spleen lymphocytes. Macrophages were found to contain about 18 times higher ANAE activity than mature lymphocytes.

Characterization of the Juvenile Hormone Esterases During Embryogenesis of the House Cricket,Acheta domesticus

Summary Juvenile hormone (JH) and α-naphthyl acetate (α-NA) esterase activity was measured on a daily basis during embryogenesis of the house cricket, Acheta domesticus. In eggs dissected from the lateral oviducts and embryos through blastokinesis, there were elevated levels of nonspecific JH esterase activity. The JH esterase activity could not be resolved from the α-NA esterase activity by gel filtration chromatography and the metabolism of both substrates was inhibited equally by 0,0-diisopropyl phosphorofluoridate (DFP). From blastokinesis through egg hatch, the JH esterase activity was maintained at relatively low levels and was resolved from the α-NA esterase activity by gel filtration. The α-NA esterase activity was inhibited by DFP while the JH esterase activity was relatively unaffected. Low JH titers in eggs must be maintained through blastokinesis for normal development. Elevated JH esterase activity in eggs during this period appears to have a functional role in the metabolism of maternal JH i...

Synergism of tumor necrosis factor and interferon-γ in induction of differentiation of human myeloblastic leukemic ML-1 cells

Natural or recombinant human tumor necrosis factor (TNF) induced NBT-reducing activity of ML-1 cells in a dose-dependent manner. Interferon-γ (IFN-γ) induced NBT-reducing activity only marginally. However, when IFN-γ was combined with TNF, induction of NBT-reducing activity was remarkably increased. IFN-α or -β had almost no effect on the induction of NBT-reducing activity of ML-1 cells, either alone or in combination with TNF. Treatment with both TNF and IFN-γ synergistically enhanced morphological changes, growth inhibition and activity of Fc receptors, and NBT reduction in ML-1 cells, but not phagocytic activity. The TNF treated cells were classified as macrophage-like by morphology, and by lineage-specific α-naphthyl acetate esterase stain. The results indicate that combinations of TNF and IFN-γ act synergestically in the induction of differentiation of human myeloblastic ML-1 cells.

Staining of pig lymphocytes subpopulations with acid α-naphthyl acetate esterase

The use of α-naphthyl acetate esterase (ANAE) as a T cell marker in some other species and the broad correlation of incidence of ANAE-positivity and E rosette-formation in the pig suggest that ANAE-staining may be a T-cell marker in the pig. However, by studying the staining of lymphocytes within a variety of rosettes in fixed preparations a similar incidence of pig blood lymphocytes were found to be ANAE + among T cells (E rosettes formed in dextran), B cells antiglobulin rosettes) and Fc-γ receptor-bearing B and T cells (EA rosettes in saline and dextran): complement (C′) receptor-bearing cells showed a higher incidence of staining than other lymphocytes. Analysis of staining morphology suggested that certain morphologies within the B and T lineages may be confined to subpopulations. Thus ANAE positivity is certainly not a marker identifying blood T lymphocytes but could be of some value indicating subpopulations of B and T lymphocytes.

Nuclear segmentation, ultrastructure, and cytochemistry of blood cells from dogs with Pelger-Huët anomaly

Nuclear segmentation, ultrastructural features and cytochemical staining reactions of blood cells from 7 dogs with the heterozygous form of Pelger-Huët (P-H) anomaly were investigated. Significant nuclear hyposegmentation of granulocytes, monocytes and megakaryocytes was found, indicating a common stem cell defect in the nuclear segmentation or lobulation process. Neutrophils and eosinophils from female P-H dogs lacked sex chromatin drumsticks. Other than the suggestion of nuclear hyposegmentation and the presence of coarse heterochromatin, no ultrastructural abnormalities were discerned. Cytochemical staining by α-naphthyl acetate esterase, alkaline phosphatase, naphthol ASD chloroacetate esterase, periodic acid-Schiff, peroxidase and Sudan black B techniques gave similar results for P-H and control dog leukocytes and platelets.

Role of juvenile hormone metabolism during embryogenesis of the house cricket, Acheta domesticus

The duration of embryogenesis was 9.5 days for house crickets, Acheta domesticus (L.), reared at 35°C. The major route of juvenile hormone (JH) metabolism was ester hydrolysis. The level of α-naphthyl acetate (α-NA) esterase activity per mg wet weight remained relatively constant throughout embryogenesis and was similar to that of eggs dissected from the oviducts. JH esterase activity per mg wet weight was highest in the dissected and day-1 eggs, declined to one-third of this peak activity by day 5, and then remained unchanged through hatching. Two populations of esterases (130,000 and > 200,000 in molecular weight) which metabolized JH and α-NA were resolved in day-1 eggs by gel filtration chromatography. Specific JH esterase appeared by day 4 with a molecular weight of 200,000. Correlative evidence is presented from other insect species that supports a functional role for JH metabolism during embryo development.

Cathepsin B synthesis by the HL60 promyelocytic cell line: effects of stimulating agents and anti-inflammatory compounds

Cathepsin B synthesis by the human HL60 promyelocyte cell line was investigated by immunohistochemistry and by the assay of the enzyme in cell lysates using a fluorimetric substrate. HL60 cells were shown to produce cathepsin B in response to treatment with 12- O-tetradecanoylphorbol 13-acetate (TPA). Intracellular levels of cathepsin B and immunohistochemical staining of the enzyme were related to time in culture with increasing concentrations of TPA from 1 nmol/l to 8.0 nmol/l. Synthesis of cathepsin B was associated with TPA-induced phagocytic activity of cells in culture, expression of α-naphthyl acetate esterase and reduced cell division. Cathepsin B production was, therefore, related to differentiation of the HL60 promyelocytes into mature macrophage-like cells. Cathepsin B activity in HL60 cell lysates was significantly increased by incubation of the cells with 10 μg/ml endotoxin (lipopolysaccharide) from Escherichia coli, but not carageenan. The production of cathepsin B by TPA-induced HL60 cells was significantly reduced by 0.25 μmol/l dexamethasone and the non-steroidal anti-inflammatory compound 4-(6-methoxy-2-naphthyl)-butan-2-one but not by indomethacin. The HL60 promyelocytic cell line is a useful model for the study of factors affecting proteinase synthesis by human mononuclear phagocytes.

The distribution of N-acetyl-β-glucosaminidase, β-glucuronidase, acid α-naphthyl acetate esterase and α-naphthyl acetate esterase in subpopulations of thymocytes, bone marrow cells and other lymphoid organs in mice

1. 1. Thymocytes, bone marrow lymphocytes, as well as lymphocytes from spleen, lymphoid nodes and peripheral blood were obtained from BALB/c mice. Subpopulations of BALB/c bone marrow T-lymphocyte precursors and immature (small) and mature (large) thymocytes, as established by the percentage of terminal deoxynucleotidyl transferase (TdT) and peanut agglutinin (PNA) positive cells, were obtained by centrifugation on discontinuous density gradients. 2. 2. The activities of N-acetyl-β-glucosaminidase (NAG), β-glucuronidase (BG), acid α-naphthyl acetate esterase (ANAE) and α-naphthyl acetate esterase (NAE) were determined by enzymatic assays of cell extracts of the diverse T-lymphocyte subpopulations, in order to follow their evolution with the maturation of the T-lymphocytes in the thymus. 3. 3. These activities were compared with that determined in lymphocytes from spleen, lymphoid nodes and peripheral blood. 4. 4. The glucidases BG and NAG and the esterases ANAE and NAE present a high decrease in their activities from bone marrow T-lymphocyte progenitors to immature thymocytes. 5. 5. BG, NAG and ANAE activities undergo an about 3-fold increase with the evolution of the thymocytes from small to large cells. Whereas the level of the NAE activity decreases (2-fold) with that evolution of the thymocytes. 6. 6. Lymphocytes from spleen and lymphoid nodes exhibit activities of the glucidases and, specially, the esterases marked by higher than those of thymocyte populations. 7. 7. Peripheral blood lymphocytes also present NAG, ANAE and NAE activities higher than in thymocytes, but their BG activity is lower.

Esterase-positive inflammatory cells in human periapical lesions

The relative distribution of acid α-naphthyl acetate esterase staining cells was studied in inflammatory infiltrates of acute (rapidly progressive) and chronic (slowly progressive) inflammatory periapical lesions, selected on the basis of clinical data. Differential cell counts on sections from the acute (10 granulomas) and chronic (3 granulomas and 7 cysts) inflammatory lesions showed that plasma cells were the predominating α-naphthyl acetate esterase-reactive cells (27%) in acute lesions, whereas T lymphocytes predominated in chronic lesions (25%). Macrophages were more abundant in chronic lesions (13%) compared with acute lesions (6%). Large granular lymphocytes (natural killer cells) could not be identified with certainty among the α-naphthyl acetate esterase-positive cells. The results indicate that there is a relationship between the composition of the inflammatory cell infiltrate and the progression of an inflammatory periapical lesion. The relative distribution of acid α-naphthyl acetate esterase staining cells was studied in inflammatory infiltrates of acute (rapidly progressive) and chronic (slowly progressive) inflammatory periapical lesions, selected on the basis of clinical data. Differential cell counts on sections from the acute (10 granulomas) and chronic (3 granulomas and 7 cysts) inflammatory lesions showed that plasma cells were the predominating α-naphthyl acetate esterase-reactive cells (27%) in acute lesions, whereas T lymphocytes predominated in chronic lesions (25%). Macrophages were more abundant in chronic lesions (13%) compared with acute lesions (6%). Large granular lymphocytes (natural killer cells) could not be identified with certainty among the α-naphthyl acetate esterase-positive cells. The results indicate that there is a relationship between the composition of the inflammatory cell infiltrate and the progression of an inflammatory periapical lesion.

Juvenile hormone esterase activity in developmentally synchronous ultimate stadium larvae of the migrant insect, Anticarsia gemmatalis

Growth and development of cumulatively aged velvetbean caterpiller ( Anticarsia gemmatalis Hübner) larvae were characterized on the basis of weight, stadium and gate. Criteria were developed for the selection of synchronous ultimate (5th)-stadium larvae before they entered the ultimate stadium. Using synchronous larvae, precise determinations were made of haemolymph juvenile hormone III esterase activity, α-naphthyl acetate esterase activity and protein concentration. The first of two juvenile hormone III esterase activity peaks coincided with the time of maximal weight gain while the second peak coincided with formation of the prepupa. Juvenile hormones (5 × 10 −6 M) I, II and III were hydrolyzed at similar rates. The α-naphthyl acetate esterase activity and protein patterns differed from the pattern of juvenile hormone III esterase activity. A major peak of α-naphthyl acetate esterase activity was observed during the wandering phase of the penultimate and ultimate stadia. Haemolymph protein concentration increased to a peak, which coincided with the initiation of wandering, and then decreased through pupation.

Inhibition of Penicillium duponti carboxylesterase by the fungicides captan and folpet

Captan, folpet, and perchloromethylmercaptan were effective inhibitors of Penicillium duponti p-nitrophenylpropionate esterase activity (I 50 = 0.5 – 2 μ M) whereas α-naphthyl acetate esterase activity was not affected by the presence of these compounds. Captan and folpet are both equally effective at pH 7.3 and 8.3. The ionic composition of the medium had strong effects on the degree of inhibition produced by all inhibitors but did not alter esterase activity. Neither succinamide nor phthalimide caused inhibition of the p-nitrophenylpropionate esterase activity: The trichloromethylmercaptan portion of these fungicides appears to be responsible for the observed inhibition. The rapidity of captan and folpet inhibition of esterase activity (complete in < 1 min) compared to the rates of spontaneous decomposition ( t 1 2 > 1 min ) and the insensitivity of captan and folpet inhibition to hydrogen ion concentration suggest that generation of spontaneous decomposition products is not required for inhibition. The results are consistent with a mechanism in which the entire fungicide molecule binds to the protein followed by enzyme-promoted reactions of captan and folpet which result in loss of esterase activity.

Alternative myelomonocytic differentiation of HL-60 reflects dual prospective potency of promyelocytes in human

The permanent promyelocytic cell line HL-60 was subjected to stimulation with dimethyl sulfoxide (DMSO) and retinoic acid (RA), as well as 12- O-tetradecanoylphorbol-13-acetate (TPA) and lymphokine conditioned media for the induction of granulocytic or monocytic differentiation, respectively. Cells were investigated cytochemically using α-naphthylacetate esterase (acid esterase; AcE), naphthol AS-D chloroacetate esterase, and peroxidase reactions. In addition, the granulocyte or monocyte specific isoenzyme patterns of AcE as an intracytoplasmic property and the immunoreactivity to monoclonal antibodies recognizing granulocytes and monocytes (Ki-M2, Ki-M5) or monocytes alone (Ki-M1) were considered. The results indicated that HL-60 cell line bear the potency to evolve into granulocytes as well as monocytes. Additional studies performed on normal human bone marrow stained for AcE led to the conclusion that the myeloid cell line remains bipolar until the maturation stage of promyelocytes. Myelocytes being AcE positive only in 11.5 ± 5.0 are heterogeneous and display the first indications of separated monocytic or granulocytic differentiation.

Detoxication capacity in the honey bee, Apis mellifera L

Various detoxifying enzymes, including microsomal oxidases, glutathione S-transferases, esterases, epoxide hydrolase, and DDT-dehydrochlorinase, were assayed in adult worker bees ( Apis mellifera L.) using midguts as the enzyme source. A cell-free system was used for all enzyme assays, except that microsomal oxidases required intact midgut because of the inhibitor encountered. Midgut microsomal preparations contained mainly cytochrome P-420, the inactive form of cytochrome P-450, which may explain the low microsomal oxidase activity in microsomes. All enzymes studied were active, suggesting that the high susceptibility of honey bees to insecticides is not due to low detoxication capacity. Sublethal exposure of honey bees to various insecticides had no effect on these enzyme activities, with the exception of permethrin which significantly stimulated the glutathione S-transferase, and malathion, which significantly inhibited the α-naphthylacetate esterase and carboxylesterase.

Substituted thiotrifluoropropanones as potent selective inhibitors of juvenile hormone esterase

A series of 27 substituted thio-1,1,1-trifluoropropanones was synthesized by reacting the corresponding thiol with 1,1,1-trifluoro-3-bromopropanone. The resulting sulfides were screened as inhibitors of hemolymph juvenile hormone esterase and α-naphthyl acetate esterase activity of the cabbage looper, Trichoplusia ni, electric eel acetylcholinesterase, bovine trypsin, and bovine α-chymotrypsin. The presence of the sulfide bond increased the inhibitory potency on all of the enzymes tested when compared with compounds lacking the sulfide. In general, the compounds proved to be poor inhibitors of chymotrypsin and moderate inhibitors of trypsin. By varying the substituent on the sulfide, good inhibitory activity was obtained on α-naphthyl acetate esterase, acetylcholinesterase, while some of the compounds proved to be extremely powerful inhibitors of juvenile hormone esterase. The most powerful inhibitor tested was 3-octylthio-1,1,1-trifluoro-2-propanone, with an I 50 of 2.3 × 10 −9 M on JH esterase. This compound showed a molar refractivity similar to that of the JH II backbone, was not toxic to T. ni, and was moderately toxic to mice, with a 48-hr LD 50 of >750 mg/kg. It effectively delayed pupation when applied to prewandering larvae of T. ni, as expected for a JH esterase inhibitor. Thus, some members of this series are promising for evaluating the role of JH esterase in insect development. The series also indicates that, by varying the substituent on the sulfide moiety, potent “transition-state” inhibitors can be developed for a wide variety of esterases and proteases.

Temporal variations of hemolymph esterase activity and juvenile hormone titers during ovocyte maturation in Acheta domesticus (Orthoptera)

Using in vitro methods, juvenile hormone (JH) esterase activity and α-naphthylacetate esterase activity were determined in the hemolymph during the first reproductive cycle of the house cricket, Acheta domesticus. Biochemical properties of the hemolymph JH esterase were studied. α-Naphthylacetate esterases increased during the first gonotrophic cycle:peaks of their activity could be observed concomitant with peaks of JH esterase activity. The fluctuations in JH esterase activity correlated with those of hemolymph JH titers. The results are discussed.

Inhibition of non-specific leukocyte esterase activity. Absence of monocyte esterase activity due to phosphoric and thiophosphoric acid ester intoxication

In vitro evaluation of the effect of five insecticidal phosphoric and 11 thiophosphoric acid esters on different, non-specific human leukocytes esterases indicated that most of the organic phosphor compounds studied inhibited the activity of neutral α-naphthylacetate esterase, α-naphthylbutyryl esterase, and naphthol AS acetate esterase, i.e. the monocyte esterases. The extent of inhibition was dose dependent; the inhibiting dose being identical for the various non-specific esterases. Reactivation with Obidoxim was not successful. Monocyte esterase activity in a human survivor of E 605 intoxication was detectable only after serum acetylcholinesterase had returned to normal levels. The organic phosphor compound studied, however, inhibited neither acid α-naphthylacetate esterase nor naphthol AS-D chloroacetate esterase activity.

Blast Crisis of Chronic Myelogenous Leukemia with Tumor Formation Characterized by T-cell Features—A Case Report

We report a case of chronic myelogenous leukemia (CML) associated with pronounced peripheral lymphadenopathy, with the cells having the philadelphia (Phl) chromosome and T-cell features. A 23-year-old man who was diagnosed as having CML and treated with busulfan was admitted to our hospital because of increasing hepatosplenomegaly and pronounced lymphadenopathy. An axillary lymph node biopsy disclosed that the malignant cells formed rosettes with neuraminidase-treated sheep red blood cells (En) (95.0%) and were positive for Leu 1 (91.8%). Of the cytochemical reactions, peroxidase was negative and periodic acid-Shiff, acid alpha-naphthyl acetate esterase and beta-glucuronidase were all positive. The karyotype of the bone marrow cells was 46 XY Phl positive (22q-), and that of the lymph node cells was 51 XY Phl positive +8, +9, +18, +19, +21, 22q-. He was treated with various anti-leukemic agents and irradiation. Despite such treatments, he died of pneumonia. This is a report of a CML patients with blast crisis and tumor formation characterized by T-cell features.