High Acid Phosphatase Activity Research Articles

Development of malignant fibrous histiocytoma induced by 7,12-dimethylbenz[a]anthracene in the rat: characterization of early atypical cells.

Morphologically atypical cells were first detected in the adjacent connective tissue 98 days after implanting a paraffin pill containing 2 mg of 7,12-dimethylbenz[a]anthracene (DMBA) into the subcutaneous tissues of rats. These cells subsequently formed groups and finally produced gross malignant fibrous histiocytomas (MFH). Early atypical cells were located between proliferating fibroblasts and histiocytes in the center of a fibrous capsule surrounding the DMBA pill. They exhibited a smooth cell surface, dilated rough endoplasmic reticulum, multiple Golgi complexes, and were often associated with newly formed collagen. These cells incorporated [3H]thymidine and [3H]proline intensively, and showed weak acid phosphatase activity but no features diagnostic of macrophages (microvilli, numerous lysosomes, high acid phosphatase and non-specific esterase activities, antigens recognized by monoclonal antibodies ED1 and OX-42 and vital staining with trypan blue). There was no evidence that atypical cells differentiated into muscle cells (no expression of desmin or the alpha-sarcomeric form of actin) or Schwann cells (no expression of S-100 protein). No point mutation in the neu gene at nucleotide 2007, specific for N-ethyl-N-nitrosourea- and DMBA-induced malignant rat schwannomas, was detected by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analyses. These results support the view that malignant fibrous histiocytoma is derived from immature fibroblasts exhibiting pronounced phenotypic diversity during the later stages of carcinogenesis.

Role of alveolar macrophages in precipitation of mineral elements inhaled as soluble aerosols.

The lysosomes of several varieties of cells such as the tubular proximal cell of the kidney and the alveolar macrophage have the ability to concentrate and precipitate several elements inhaled in water-soluble form, usually as phosphate. The mechanism involved is attributed to the high acid phosphatase activity of lysosomes and can be considered as an in vivo Gomori reaction. Among the elements studied, most of them are chemotoxic or radiotoxic (Cr; group IIIA: Al, Ga, In; rare earths: La, Ce, Tm; actinides: Th, U). In the lung macrophage, this mechanism of intralysosomal concentration and precipitation may prevent the diffusion of these toxic elements through the alveolar membrane.

The pituitary-thyroid axis effects on ocular and orbital tissues: a histological, histochemical, and morphometric study.

This work tests the notion that the effect of thyroid hormone on orbital and ocular tissues is mediated through its action on their lysosomal enzymes. Hyperthyroidism was produced in guinea pigs by thyroxin and TSH; hypothyroidism was induced by thiouracil. After treatment for 10 to 21 days, several ocular and orbital tissues were taken for histological, morphometrical, and histochemical examinations. High acid phosphatase activity was demonstrated in extraocular muscles, optic nerve and the retinal pigment epithelium of thyroxine- and TSH-treated animals. The findings fit the notion that the effects of thyroid hormones are mediated through lysosomes also in ocular and orbital tissues.

Functional micro-anatomy of the digestive gland of the scallopPecten maximus(L.)

The digestive gland of Pecten maximus consists, as in other lamellibranchs, of numerous blindending tubules which communicate with the stomach by partially ciliated main ducts and non ciliated secondary ducts. The non-ciliated cells of the main ducts are characterized by a well developed brush border constituted by high and dense microvilli and a strong pinocytotic activity. Ciliated and non ciliated cells have a very similar fine structure. The digestive tubules have a large lumen and contain digestive cells at different stages of absorption, digestion and excretion, one part of the tubules being functional while the other is disintegrating. The dark crypts contain the flagellated secretory cells, characterized by a well developed granular endoplasmic reticulum, and the young immature cells which may replace both the secretory and digestive cells. The numerous lipid droplets occurring in the digestive duct cells and in the digestive cells reveal the lipid storage function of the digestive gland. Several enzyme activities involved in digestion have been localized in the digestive gland. High amylase activity and cellulase and lysozyme activities have been found in the ducts and in the tubules, whereas no proteolytic activity could be detected histochemically. Some intracellular peptidases and glycosidases have been localized in the cells of the digestive gland, especially in the brush-border cells of the ducts and in the functional part of the tubules. High alkaline and acid phosphatase activities are displayed by the duct brush-border cells and the digestive and secretory cells. These results show the main role of the digestive gland, both in extracellular digestion (secretion of the digestive enzymes) and in absorption and intracellular digestion and provide information on the respective functions of the different cells within these processes.

Japanese quail and human acid maltase deficiency: A comparative study

A morphological study was carried out on the skeletal muscle of a mutant Japanese quail with acid maltase deficiency (AMD). The affected quails began to experience difficulty in lifting their wings about 6 weeks after hatching. Four weeks after hatching, before symptoms appeared, alpha-1, 4-glucosidase activity in skeletal muscle was decreased to less than 10% of the control level, and muscle fibers possessed many vacuoles containing periodic acid Schiff (PAS) positive material which was digested by diastase, and showed high acid phosphatase activity. Although both red and white muscles were involved, the pectoralis superficialis (PS, white) muscle was preferentially affected, showing intracytoplasmic vacuoles, variation in fiber size and fatty tissue replacement relatively early in the disease. The quails' disease closely resembled human late onset AMD in the slow clinical course, the presence of residual acid alpha-glucosidase activity and the muscle pathology. This mutant quail seems a useful model for elucidation of the muscle degeneration in human AMD (glycogen storage disease type 2).

Properties and intracellular distribution of a cathepsin D-like proteinase active at the acid region of Musca domestica midgut

Musca domestica larval midgut display in cells and luminal contents a proteolytic activity with a pH optimum of 3.0–3.5. This activity is abolished by pepstatin and is insensitive to soybean trypsin inhibitor and to sulfhydryl proteinase inhibitors. The acid proteinase occurs in multiple forms with M r values in the range 40,000–80,000 and with pI values of about 5.5. The proteinase inactivates at 60°C according to apparent first-order kinetics and Lineweaver-Burk plots of its activity against albumin concentration are rectilinear, suggesting that the multiple forms have similar properties. The proteinase reacts slowly with diazoacetylnorleucine plus CuSO 4, is stable in alkaline media, is inhibited by dithiothreitol, hydrolyses hemoglobin better than albumin and is virtually not active upon synthetic substrates for pepsin. These properties are similar to those of cathepsin D. The specific activity of the acid proteinase determined by titration with pepstatin is 680 units/mg of proteinase and the K D of the pepstatin-proteinase complex is 1.5 nM at 30°C. The acid proteinase occurs mainly in midgut subcellular fractions characterized by a high specific activity of molybdate-inhibited acid phosphatase and a large number of secretory-like vesicles. It is proposed that the M. domestica midgut acid proteinase is a cathepsin D-like proteinase evolved to function in luminal contents. The lack of ATP activation of the midgut enzyme supports this hypothesis, since ATP is thought to regulate cathepsin D-proteolysis inside lysosomes.

Prostatic Acid Phosphatase in Serum of Patients with Prostatic Cancer Is a Specific Phosphotyrosine Acid Phosphatase

We developed an assay to measure at acid pH the phosphotyrosine phosphatase activity in sera from patients with prostatic cancer. The method used quantifies the inorganic phosphate liberated from phosphotyrosine after incubation with serum, followed by the deproteinization of the reaction mixture. A high acid phosphatase (EC 3.1.3.2) activity towards phosphotyrosine was observed in all sera from patients with increased activity of prostatic acid phosphatase. This activity represented 96% of prostatic acid phosphatase and 77% of total acid phosphatase activities. Moreover, it was correlated (r = 0.91) with the amount of serum prostatic acid phosphatase determined by radioimmunoassay. When serum acid phosphatase activity was measured on several phosphorylated substrates, preferential hydrolysis was demonstrated for those in which the phosphate group was esterified on an aromatic ring rather than those presenting an aliphatic chain. Among phosphoamino acids, only phosphotyrosine was a good substrate, with little or no activity observed with phosphoserine and phosphothreonine. Human seminal plasma and partially purified prostatic acid phosphatase, tested for their activity on some of these substrates, gave similar results. On the other hand, sera from patients with above-normal alkaline phosphatase activity and no prostatic disease showed little or no activity on phosphotyrosine at both acid and alkaline pH values. Evidence is presented that the prostatic acid phosphatase in serum is a specific phosphotyrosine acid phosphatase.

Enzyme activity in soils showing enhanced degradation of organophosphate insecticides

Studies were conducted to determine whether soils that showed enhanced biodegradation of organophosphate insecticides had significantly different enzyme activities from those in the same soils with no previous exposure to the insecticides. Twenty-one pairs of soils were collected from farms in the Midwest where chlorpyrifos, terbufos, fonofos, or phorate had failed to protect corn (Zea mays L) from corn rootworm (Diabrotica sp). Each soil was analyzed for acid and alkaline phosphatase, phosphodiesterase, phosphotriesterase, and dehydrogenase activities. Over 40% of the insecticide-treated soils had higher acid phosphatase activity than the fence row soils which had no previous exposure to the insecticide. Over twothirds of the soils treated with fonofos had higher acid phosphatase and phosphotriesterase activity than the fence row soils. If these enzymes are not directly involved in the biodegradation of the insecticitde, they may be indicative of enhanced biodegradation and may be used to predict which soils may be prone to insecticide failure.

Acid phosphatase activity in roots of scots pine (Pinus sylvestris L.) seedlings fertilized with different nitrogen sources

Cell wall-associated acid phosphatase activity was analysed in Scots pine roots treated with different nitrogen sources (urea, ammonium tartrate, calcium nitrate) supplied at different levels. The highest activity of acid phosphatase was found in pine roots supplied with ammonium tartrate and was associated with the highest growth of pine roots.

Distribution of acid phosphatase and ?-glucuronidase in the hypoplastic cerebellum of jaundiced Gunn rats

Activities of acid phosphatase and beta-glucuronidase in the cerebella of young jaundiced (j/j) and non-jaundiced (j/+; control) Gunn rats were studied with the enzyme histochemical method. The cerebellum of j/+ rats showed high acid phosphatase activities in Purkinje cells and neurons in the cerebellar nuclei. In j/j rats, a number of neurons were lost and numerous microglialike cells with a high acid phosphatase activity appeared in the hypoplastic cerebellum. Although beta-glucuronidase activity was rarely detected in the control cerebellum, a high enzyme activity was observed associated with microglialike cells in j/j rats. The present results provide a cytological basis for the reported differential increase in the activities of these lysosomal enzymes in the j/j rat cerebellum.

The effect of exogenous phosphate deficiency on the activity of acid phosphatase of the root of two maize genotypes

In two maize genotypes, the effect of exogenous phosphate deficiency on acid phosphatase activity of the apical part of primary root was followed in dry and imbided grains. Higher acid phosphatase activity was found in the genotype LG 5. The enzyme activity increased after 18 h grain imbibition in the two genotypes. After 48 h germination no differences were found between the genotypes. After 24 h cultivation of root segments in a solution of 0.25 mM CaSO4.2H2O,0.1 mM MgSO4.7H2O, and further after 3,6 and 24 h cultivation in solutions with and without phosphate genotypic differences were found in acid phosphatase activity as well as in dry mass production. An increase in enzymatic activity due to exogenous phosphate deficiency was registered in the two genotypes only after 24 h cultivation.

Acid phosphatase, alkaline phosphatase, and nitrate reductase activity of selected ectomycorrhizal fungi

Seventeen isolates, encompassing five genera and eight species of ectomycorrhizal fungi, were compared for acid phosphatase, alkaline phosphatase, and nitrate reductase activity. Isolates within species differed in enzyme activity and isozyme patterns by host specificity and site (as exemplified by the genus Suillus). Host and site may have affected phosphatase enzyme activity. Generally, the Douglas-fir associates, which dominate in mesic sites, have higher acid phosphatase activity than pine associates, which mostly occupy xeric sites; however, pine associates from mesic sites also have higher acid phosphatase activity (e.g., S. tomentosus). In four isolates of Amanita muscaria, the effect of site was also apparent. Two of them, which have significantly higher acid phosphatase activity than the others, were isolated from mesic sites. The isozyme pattern of the genus Suillus appeared to be separated by host groups. Other isolates with only one species also differed more or less by host groups. They shared at least one band within host groups, except for the two isolates of Paxillus involutus from different hosts. The P. involutus S-403 isolated from an orchard showed much higher nitrate reductase activity than all other isolates. No apparent differences in nitrate reductase activity were found between the other isolates.

Underfeeding and exposure to short photoperiod alters rat pineal and Harderian gland lysosomal enzyme activities.

Harderian gland (HG) weight and lysosomal enzyme activity were evaluated after 21-day-old female rats were singly caged in a long (LP; 14:10 LD) or short (SP; 8:16 LD) photoperiod and fed on one of two dietary regimens (fed ad libitum or 50% underfed) for 50 days; an additional fed and an underfed group of animals in LP were injected every afternoon with 100 micrograms melatonin. Absolute HG weights were significantly lower in all underfed groups compared to their respective fed controls or to the LP fed control group. Absolute HG weights of underfed rats in SP were significantly lower than the underfed rats in LP. Relative HG weights (mg/100 g body wt) were significantly higher in the underfed saline or melatonin-treated groups compared to their respective fed controls; however, HG of the underfed SP group were not different from SP-fed controls. No significant differences in HG acid phosphatase, hexosaminidase, and beta-glucuronidase activities were observed in any of the treatment groups maintained in LP. Acid phosphatase, hexosaminidase, and beta-glucuronidase activities were significantly elevated in HG of underfed animals maintained in SP compared to their respective fed controls or to the LP-underfed group. Both the underfed control and the underfed-melatonin treated groups had lower pineal protein values than their respective fed groups; underfed animals in 8:16 LD had similar pineal protein values compared to those of the fed control group in SP. Significant effects of photoperiod and underfeeding with no interaction between these variables were observed on pineal acid phosphatase. The fed group maintained in 8:16 LD had significantly higher acid phosphatase activity than the fed group kept in 14:10 LD. In conclusion, underfeeding resulted in severely reduced body weights and absolute Harderian gland weights. Increased activity in certain lysosomal enzymes occurred in both the pineal and Harderian gland and in some instances this was dependent upon the light cycle and dietary regimen to which the animals were exposed.

Selection of amylolytically active Aspergillus niger mutants to 2-deoxy-D-glucose

As a result of mutagenization and passaging on 2-deoxy-D-glucose containing medium, 10 Aspergillus niger strains resistant to this agent were obtained. These showed (with one exception) an increase in the activity of glucoamylse, the level of which ranged widely in individual cases from several to over 200% in comparison with the parent strain. A weaker rate of glucose accumulation in derepressed strains may account for the fact that the mechanism of their resistance to deoxyglucose is connected with disturbance of the system of glucose transport. However, it is possible that a high activity of acid phosphatase, which the obtained deoxyglucose-resistant cultures showed, may be involved here. Apart from the biochemical character of the catabolic derepression, it seems that it can already be successfully utilized to increase the productivity of industrial mould cultures.

Acid phosphatase activity in maize leaves as related to their evolution and phosphorus deficiency

The effect of phosphorus deficiency on the activity of acid phosphatase of the first, second and third leaves of maize plants was followed. The supernatant obtained by centrifuging the homogenate of plant tissue at 1500 ×g was further centrifuged at 18 000 ×g, the sediment marked as fraction II and the supernatant as fraction III. Acid phosphatase activity of fraction II of the first to third leaves was for the whole period of culture higher in plants grown in the nutrient solution without phosphate. In fraction III this relation was established in the first leaf, after 3 days of culture in the second leaf and after 5 days in the third leaf. In all leaves higher enzyme activity was unambiguously determined in fraction III when compared with fraction II. Higher acid phosphatase activity was established in those leaves which were younger in their development, particularly in the first days of culture. With the ageing of leaves the enzyme activity decreased.

Phosphatase and peptidase activities in reindeer antler throughout the growth cycle

Abstract Alkaline phosphatase activity increased 10,000‐fold over basal levels in reindeer antler during the period of massive growth. The activity was highest in the zone of differentiating chondrogenic cells in the growing tip. The activity in the shaft was lower. PPiase activity was also highest in the antler tip, but compared to alkaline phosphatase, was more tightly membrane‐bound. Ca2+, Mg2+‐ATPase activity was not so markedly elevated in the tip as were the other activities. During the growth cycle, the highest activities of acid phosphatase and amino peptidases occurred later than the peak of alkaline phosphatase activity, and were found mainly in the dermis of the shaft.

Enzymhistochemische Untersuchungen an der Schweineplacenta II. Histotopik von enzymen in den areolären placentaepithelien

In porcine areolar placental epithelia, the following enzymes were demonstrated by histochemical methods after 30, 58, 80, 100, and 110 d of pregnancy, respectively: beta-N-acetyl-hexosaminidase, beta-galactosidase, beta-glucuronidase, alpha-mannosidase, acid phosphatase, alkaline phosphatase, nonspecific esterases, cytochrome oxidase, 5-nucleotidase, leucine aminopeptidase, adenosine triphosphatase, diaphorases (NADH, NADPH), glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, succinate dehydrogenase, isocitrate dehydrogenase (NAD, NADP), beta-hydroxybutyrate dehydrogenase, glycero-3-phosphate dehydrogenase, NAD-glycero-3-phosphate dehydrogenase, glutamate dehydrogenase (NAD, NADP), lactate dehydrogenase. The results show that the enzyme activities remained almost unchanged during the period of investigation. Of the dehydrogenases, the diaphorases as well as succinate and lactate dehydrogenase demonstrated generally an intensive activity within the epithelia. The activity of the other dehydrogenases was only low. The activity of unspecific esterase was very intensive within the uterine epithelia but remarkably low within chorionic epithelia. Contrarily, the reaction of adenosine triphosphatase was more intensive within chorionic than uterine epithelia. All investigated glucosidases reacted distinctly positive within chorionic epithelia, but only beta-N-acetyl-hexosaminidase and beta-galactosidase in uterine epithelia. The high activity of acid phosphatase, especially within the chorionic epithelium, seems to be connected with uteroferrin, an iron-binding protein. The histochemical results are discussed in context with the function of the areolae in histiotrophic nutrition and iron transport.

Ultrastructural and histochemical studies of granular-cell ameloblastoma.

Four cases of granular-cell ameloblastoma were studied by light and electron microscopy, histochemistry and electron enzyme histochemistry. Microscopically, granular cells commonly occurred in a follicular pattern, but, in one case, they were in a plexiform pattern. The high activity of acid phosphatase and electron microscopic features revealed that the cytoplasmic granularity was ascribed to the high content of lysosomes. It was speculated that granular-cell ameloblastoma occurs due to the altered dysfunction of tumor cells, and the age factor is related to the pathogenesis of this tumor.

Characterization of the vacuolar ATPase activity of the crassulacean-acid-metabolism plant Kalanchoë daigremontiana. Receptor modulating.

Plants performing crassulacean acid metabolism show a large nocturnal accumulation of malic acid in the vacuole of the photosynthetic cells. It has been postulated that an H+-translocating ATPase energizes the transport of malic acid across the tonoplast into the vacuole. In the present work we have characterized the ATPase activity associated with vacuoles of the crassulacean-acid-metabolism plant Kalanchoë daigremontiana and compare it with other phosphohydrolases. Vacuoles were isolated by polybase-induced lysis of mesophyll-cell protoplasts. The vacuoles had a high activity of unspecific acid phosphatase (pH optimum 5.3). The acid phosphatase was strongly inhibited by ammonium molybdate (with 50% inhibition at about 0.5 mmol m-3), but was not completely inhibited even at much higher ammonium-molybdate concentrations. In contrast, the vacuolar ATPase activity, assayed in the presence of 100 mmol m-3 ammonium molybdate, had a pH optimum of 8.0. ATP was the preferred substrate, but GTP, ITP and ADP were hydrolyzed at appreciable rates. The mean ATPase activity at pH 8.0 was 14.5 nmol h-1 (10(3) vacuoles)-1, an average 13% of which was attributable to residual acid-phosphatase activity. Inorganic-pyrophosphatase activity could not be demonstrated unambiguously. The vacuolar ATPase activity was Mg2+-dependent, had an apparent Km for MgATP2- of 0.31 mol m-3, and was 32% stimulated by 50 mol m-3 KCl. Of the inhibitors tested, oligomycin slightly inhibited the vacuolar ATPase activity and diethylstilbestrol and NO-3 were both markedly inhibitory. Dicyclohexylcarbodiimide and tributyltin were also strongly inhibitory. Tributyltin caused a 50% inhibition at about 0.3 mmol m-3. This is taken as evidence that the vacuolar ATPase might function as an H+-translocating ATPase. It is shown that the measured activity of the vacuolar ATPase would be of the right order to account for the observed rates of nocturnal malic-acid accumulation in K. daigremontiana.

Regional and subcellular distribution in mammalian brain of the enzymes producing adenosine.

The activities of 5'-nucleotidase, 2'-nucleotidase, alkaline phosphatase, and acid phosphatase were measured in rat and autopsied human brains. The four phosphatases in the rat brain showed little change in activity after death. The activities of adenosine-producing enzymes were compared in various parts of rat and human brains. When phosphatase activity was measured at pH 7.5, 5'-nucleotidase showed the highest activity in the most parts of the brain. The activity of 2'-nucleotidase and that of nonspecific phosphatase were almost the same at pH 7.5. However, higher phosphatase activity was observed in all parts of the brain when nonspecific phosphatase activity was measured at pH 10.0 or 5.5. High specific activity of 5'-nucleotidase in the brain was detected in the membranous components, especially in the synaptic membranes. The activity of 2'-nucleotidase was distributed in the soluble and synaptosomal fractions. The highest activity of both alkaline and acid phosphatases was recovered in the crude mitochondrial fraction, with the highest specific activity in the microsomal fraction. Phosphatase activity was distributed widely in the rat brain. The activity of 5'-nucleotidase was high in the medulla oblongata, thalamus, and hippocampus, but low in the peripheral nerve, spinal cord, and occipital lobe. The activity of 2'-nucleotidase was high in the vermis and frontal lobe. The highest acid and alkaline phosphatase activities were detected in the frontal lobe and in the olfactory bulb, respectively. The distribution of the four phosphatases in the autopsied human brain was similar to that in the rat brain. The highest 5'-nucleotidase activity was observed in the temporal lobe and thalamus.(ABSTRACT TRUNCATED AT 250 WORDS)