Total Acid Phosphatase Research Articles

Blood biochemical profile of the South American rattlesnake (Crotalus durissus terrificus) in captivity

Blood samples were collected from 180 healthy specimens of the South American rattlesnake, Crotalus durissus terrificus, in captivity. All animals were in good clinical condition. Normal biochemical reference values were established for the following: total proteins, albumin, globulins, uric acid, creatinine, urea, glucose, triglycerides, cholesterol, total lipids, calcium, phosphorus, potassium, sodium, chloride, magnesium, GOT (AST), GPT (ALT), and alkaline phosphatase (ALP). Samples were obtained by venipuncture of the ventral tail vein. Values were compared with published data for Boidae, Elapidae, and Viperidae.

Analysis of Arylsulfatases A and B, Acid Phosphatase, Lactate Dehydrogenase, and Aspartate Transaminase in Chronic Periapical Lesions of Endodontic Origin

Attempts were made to detect and measure the activities of arylsulfatases. A&B acid phosphatase, lactate dehydrogenase, and glutamate oxaloacetate transaminase (aspartate transaminase) enzymes in human chronic lesions of endodontic origin. Thirteen periapical lesions of endodontic origin and 11 noninflamed control periapical tissues were obtained. The specimens were carried to the laboratory on liquid nitrogen and kept at -70 degrees C. Samples were thawed, homogenized, and then assayed for enzyme activities. The specific activities of arylsulfatase A (nmol/hr/mg protein) were 55.0+/-10.7 (chronic lesions) vs. 3.4+/-2.2 (controls) (p < 0.01). Arylsulfatase B specific activities (nmol/hr/mg protein) were 50.3+/-6.4 (chronic lesions) vs 91.8+/-18.4 (controls). Total acid phosphatase activities (mU/mg protein) were 45.8+/-6.6 (chronic lesions) vs. 26.8+/-3.1 (controls). Lactate dehydrogenase activities (Berger-Broida units/mg protein) of the chronic periapical lesions were significantly higher than the control group (362+/-63.2) vs. (140+/-46.0) (p < 0.05). There was no significant difference between the specific activities of aspartate transaminase in chronic lesions and the control group (68.0+/-14.5) vs. (53.0+/-10.4) mU/mg protein).

Kinetic determination of acid phosphatase activity by double injection flow analysis with electrochemical detection

A two-points kinetic procedure has led to an enzyme assay for the measurement of total and prostatic acid phosphatase (PAP) activity by flow injection analysis with electrochemical detection. A double injection valve is employed for the simultaneous injection of the substrate ( α-naphthyl phosphate) and enzyme. Successive injections are directed into different reaction coils for amperometric measurement of the α-naphthol produced at different times. Total and prostatic activity is determined in control sera and results were compared with those obtained spectrophotometrically.

Determination of acid phosphatase activity in a double injection flow system with electrochemical detection

An enzyme assay for the measurement of total and prostatic acid phosphatase activity by flow injection analysis with electrochemical detection has been developed. The substrate (α-naphthyl phosphate) and enzyme are simultaneously injected to the system by means of a double injection valve. The α-naphtol produced after stopping the flow is amperometrically quantified. The calibration curve is linear from 0.01 to 1.3 u/l with a limit of detection of 0.006 u/l. Results were compared with that obtained spectrophotometrically.

Levels of enzymes in leukaemic mice treated withAeromonas L-asparaginase.

L-asparaginase isolated in our laboratory fromAeromonas has been found to be antileukaemic. In the present study changes in the levels of serum enzymes in leukaemic mice and under treatment withAeromonas L-asparaginase has been compared. A significant increase in the levels of serum lactate dehydrogenase with tumour growth and a decrease during therapy was observed. A significant decrease in alanine transaminase activity during tumour growth and an increase during treatment was noticed. Increased levels of aspartate transaminase and alkaline phosphatase was observed during enzyme therapy. Total acid phosphatase was found to be increased during tumour growth and decreased considerably during treatment.

Low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase in the developing chick brain: partial characterization and levels during development.

Low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase is largely expressed in chick brain tissue during development. The enzyme was purified from brain extract prepared from 19-day-old chick embryos and from adult chickens using ammonium sulfate fractionation, gel filtration on Sephadex G-75 and two DEAE-Cellulose ion-exchange chromatography steps. The purified enzymes from embryo and adult chick brains show identical molecular weight values (about 18-20 kDa) and biochemical and structural properties such as substrate specificity, sensitivity to inhibitors, and number of free reactive sulphydryl groups. These data suggest that they are the same enzyme protein. Although the total acid phosphatase activity does not change appreciably during development, the activity associated with the low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase markedly increases after birth and reaches the adult values within the first week of life. Taken together, our results suggest an involvement of the low molecular weight acid phosphatase/phosphotyrosyl protein phosphatase in postnatal development and maturation of chick brain tissue. The variations in tyrosine phosphorylation profile of chick brain polypeptides analyzed by Western blotting at the same developmental stages are also reported.

Phytase and acid phosphatase activities in extracts from roots of temperate pasture grass and legume seedlings

Phytase and acid phosphatase activities were measured in extracts from roots of 14- to 22- day old seedlings of a range of temperate pasture species that were grown aseptically in sand culture. Phytase activity from roots of phosphorus- (P-)-deficient Trifolium subterraneum L. was characterised. Activity was enhanced by 40% when extracts were passed through Sephadex G-25, and increased by a further 20–30% with the addition of either 1 mМ EDTA or 5 mМ cysteine to assay solutions. The optimum temperature for phytase activity was 50°C and the optimum pH was 5.3. When compared with phosphatase activity measured in the roots of T. subterraneum, phytase activity exhibited narrower pH and temperature optima, and was also more strongly inhibited by Co2+, Zn2+ and AsO42− ions. Significantly, for the five pasture species examined, phytase activity was less than 5% of the total acid phosphatase activity in extracts of plant roots. Measured phytase activity ranged between 0.13 and 1.7 nkat g–1 root fresh wt and was enhanced under P-deficient relative to P-sufficient growth conditions in all of the pasture species with the exception of Trifolium repens L., for which the Km constant for activity was 50% lower in P-deficient plants. When expressed on a root fresh wt basis, increases in phytase activity of ~1.25-fold were observed for extracts from T. subterraneum and Medicago polymorpha L., and of up to 3.3-fold for Danthonia richardsonii A.B. Cashmore and Phalaris aquatica L. Increases in acid phosphatase activity with P deficiency were less evident. Between 3.1% and 4.3% only of the total phytase activity measured in root extracts was eluted from intact roots into 0.1 М NaCl.

Host nuclear abnormalities and depletion of nuclear antigens induced in Trichinella spiralis-infected muscle cells by the anthelmintic mebendazole

Infection by the parasitic nematode Trichinella spiralis induces cell cycle repositioning (chronic suspension in apparent G 2/M) and genetic reprogramming in differentiated mammalian skeletal muscle cells. These changes occur in association with dramatic enlargement of infected host cell nuclei (as large as 17 μm in diameter) and nucleoli. Nuclear antigens (NA) that colocalize with host chromatin have been detected by antibodies to T. spiralis antigens, but the functions of these NA are unresolved. Mebendazole (MBZ) preferentially binds parasite versus host β-tubulins, is implicated in inhibiting secretion in nematodes and induces cytoplasmic changes in muscle cells infected with T. spiralis. These infected cell changes might be indirect via MBZ inhibition of parasite secretions. This effect would have implications for host/parasite interactions and was evaluated here. MBZ treatment of chronically infected mice caused: (1) a significant deformation of host nuclei and diminution of nucleoli by 4 and 6 days of treatment (dot), respectively; (2) a reduction of nuclear lamins A/C in infected cell nuclei that was concomitant with nuclear deformation; and (3) significant reductions in total RNA, general protein and acid phosphatase activity levels. These changes were associated with the depletion of NA from host nuclei detected by 4 dot. However, DNA content of infected cell nuclei was not detectably reduced and muscle gene expression was not reactivated. The cellular changes documented are likely to account for previously described cytoplasmic alterations induced by MBZ. Concomitant depletion of NA from infected cell nuclei suggests a role of these products in regulating nuclear functions of host cells.

Platelet-Derived Growth Factor (PDGF)-BB Stimulates Osteoclastic Bone Resorption Directly: The Role of Receptor β

The growth-promoting activity of PDGF-BB was studied on the adult osteoclasts in the present study. The PDGF receptor β was detected on the osteoclast membrane through immunohistochemistry (LSAB method) and immunomicroscopy. The PDGF-BB was exerted on the osteoclasts that adhered to the bone slice at concentrations of 0, 10, 20, 30, and 40 ng/ml. The volume of Howship's lacuna augmented significantly and the number of resorption pits also increased with its dose (p< 0.01). The activity of both total acid phosphatase (ACP) and tartrate-resistant acid phosphatase (TRAP) increased significantly. These results suggest that PDGF-BB promotes adult osteoclastic bone resorption directly through PDGF receptor β and is believed to play important roles in the bone healing process and reconstruction.

Total protein and acid phosphatase concentrations in prostatic fluid from patients with BPH compared to carcinoma.

To obtain evidence of metabolic changes in the human prostate associated with prostate pathology, in particular carcinoma of the prostate, by identifying and evaluating associated changes in prostatic secretory products. Expressed prostatic fluid (EPF) from 36 patients with carcinoma, 128 with BPH histologically confirmed, and 148 with clinical BPH was subjected to determination of protein (Lowry; UV 280 nm absorption), enzymatic (DMA modified Row procedure) acid phosphatase (AcP), and immunologically identified (Tandem-PAP immunoenzymatic assay) prostatic acid phosphatase (PAP) concentration. The important EPF findings are the following: (1) Protein concentrations (Lowry and UV determinations) are significantly increased in carcinoma as compared to histologic BPH, (2) AcP and PAP secretions remain stable in carcinoma versus BPH, and (3) AcP and PAP/Lowry protein ratios are significantly lower with carcinoma. These findings of increased protein and the decreased relative secretions of AcP and PAP to total protein (ratio) in EPF from patients with carcinoma compared to BPH support and help to characterize the diffuse metabolic alteration in the prostate associated with prostate carcinoma. EPF observations identify potential metabolic changes occurring in prostate carcinoma that may have potential clinical and investigative relevance.

Lysosomal involvement in the removal of clofibrate-induced rat liver peroxisomes. A biochemical and morphological analysis

Peroxisomal proliferators induce in rodents hepatic hyperplasia and hypertrophy; the significant increase in the peroxisomal population is accompanied by specific and reversible induction of some peroxisomal enzymes. In suckling rats born from clofibrate-treated mothers, a massive removal of proliferated organelles occurs within 3 days of recovery. In the present paper we examined the early stages of the recovery period in liver of male rats treated with clofibrate for 5 days. The lysosomal involvement in the removal of drug-induced peroxisomes was investigated under physiological conditions, ie in the absence of inhibitors of the autophagic process. Biochemical results indicate that peroxisomal β-oxidation, but not catalase activity, returns to the control values within the examined period. Total acid phosphatase activity is not affected by clofibrate treatment, but following fractionation on a linear density gradient the lysosomal marker enzyme activity is shifted towards lower density values, particularly at day 1 and 2 of recovery. This class of organelles possibly represents lysosomes involved in active autophagic processes. Acid phosphatase cytochemistry shows an increase of lysosome number at day 1 of recovery. Combination of acid phosphatase cytochemistry either with catalase cytochemistry or with catalase immunogold labelling allows to reveal organelles containing both marker enzymes. These results strongly support the involvement of autophagic processes in the removal of proliferated peroxisomes.

Conceptus, progesterone, and breed effects on uterine protein secretion in swine.

This experiment consisted of the following treatment-breed groups: 1) White crossbred gilts, 2) White crossbred gilts treated with progesterone (200 mg/d in corn oil given on d 2 and 3 after estrus), and 3) Chinese Meishan gilts. Pregnant and nonpregnant gilts (n=3 to 6) from each treatment-breed combination were assigned to be slaughtered on d 10, 11, 12, 13, and 15. At slaughter each uterine horn was flushed with 20 mL of minimal essential medium. Uterine flushings were assayed for total protein, acid phosphatase, uteroferrin, retinol-binding protein, and oxytocin. Uterine flush total protein was increased by progesterone treatment, was unaffected by pregnancy status, and was less in Meishans. Similar patterns were found for retinol binding protein and uteroferrin, except that uteroferrin was greater in pregnant than in nonpregnant gilts. Oxytocin was greater in pregnant than in nonpregnant gilts, was not influenced by progesterone treatment, and was similar in Meishan and in White crossbred gilts. These results indicate that the conceptus does not influence secretion of either total protein or retinol binding protein during pregnancy and that the onset of secretion of these uterine proteins may be controlled by progesterone. The presence of the conceptus is associated with increased uteroferrin and oxytocin production. The decreased secretion of uterine proteins in Meishan gilts may partially explain the slower embryonic development that has been reported for this breed.

Low serum testosterone and a younger age predict for a poor outcome in metastatic prostate cancer.

Carcinoma of the prostate gland is one of the most common malignancies in males. This study was undertaken to determine which factors predict the course and outcome of patients treated with first line hormonal manipulation. A total of 144 patients with Stage D2 prostate cancer who received androgen deprivation therapy were studied. Pretreatment parameters analyzed were age, performance status, analgesia usage, concurrent disease, histologic differentiation, hemoglobin, leukocyte and platelet count, serum creatinine, alkaline phosphatase, lactate dehydrogenase, prostate specific antigen, total and prostatic acid phosphatase, serum testosterone, follicle stimulating and luteinizing hormone levels, number of metastatic sites and bone scan grade. Only initial serum testosterone (> 10 nmol/l) had a positive impact on response (p = 0.0304), whereas age older than 60 years had a positive impact on time to progression (16 vs. 11 months, p = 0.0414). Both serum testosterone (26 vs. 20 months, p = 0.003), and age (28 vs. 17 months, p = 0.036) had a significant influence on overall survival. Low testosterone, indicating androgen independence, and a younger age, seem to result in a more aggressive disease and a poorer prognosis in advanced prostate cancer.

Occurrence of a community with high morbidity associated with Schistosoma mansoni infection regardless of low infection intensity in north-east Brazil

To establish the relationship between schistosome-associated morbidity and infection intensity in north-east Brazil, a parasitological and ultrasonographical study was carried out on 484 inhabitants of 4 villages (I, II, III and IV) in São Lourenço da Mata, Pernambuco, Brazil, where schistosomiasis is endemic. Quantitative stool examination using Knight's method demonstrated a high prevalence and moderate intensity of Schistosoma mansoni infection, and also that the subjects in village IV had a significantly lower prevalence and intensity of infection than those of the other villages. By ultrasonography, periportal fibrosis (PPF) and splenomegaly were found in 52% of the 299 infected subjects and 66% of the 146 infected subjects aged over 16 years old, respectively; 32% and 31% of the 299 infected subjects had abnormally high values of total bile acid (TBA) and alkaline phosphatase (ALP) activity, respectively. Liver and spleen size, PPF, and serum level of TBA and ALP were not correlated with infection intensity. There was no significant difference in the morbidity assessed by liver and spleen size, PPF, and serum analysis between the subjects in village IV and the other villages. These findings suggest the occurrence of a community with high morbidity associated with schistosomiasis regardless of low infection intensity.

Properties of secretory acid phosphatase from lupin roots under phosphorus-deficient conditions

Phosphorus deficiency induces the synthesis of acid phosphatases in roots of lupin and other plant species. In this study we examined the induction of secretory acid phosphatase (S-APase) at both the molecular and cellular levels. Lupin plants had increased levels of total acid phosphatase activity within two to five days after P was withered and levels approximately doubled by 15 days. Lateral roots and not main tap roots were responsible for this increase in acid phosphatase activity. Immunoblot analysis using antibodies raised against a purified S-APase showed that the synthesis of this protein was induced as early as 2 days in the P deficient treatment and that levels dramatically increased by 15 days. In contrast, no immunoreactive polypeptide was evident from crude extracts prepared from root tissues of—P treated plants. Immunocytochemical analysis revealed that the protein was located on the surface of epidermal cells of main tap roots and in the cell walls and intercellular spaces of lateral roots and lateral roots may actively secrete S-APase as soon as it is synthesized. A cDNA clone encoding the S-APase was isolated from a cDNA library constructed from lupin roots grown without P. The clone was 2,187 bp in length and had a single open reading frame of 637 amino acid residues. The deduced amino acid sequence was identical to the N-terminal region and peptide sequences of S-APase purified from lupin roots. A hydrophobic signal peptide region consisted of 31 amino acids. The primary structure was highly homologous to iron-zinc purple acid phosphatase from Phaseolus vulgaris (76%), secretory purple acid phosphatase from Arabidopsis thaliana (71%), and two Aspergillus phosphate repressible acid phosphatases (59% and 58%).

Acid Phosphatase in the Pathogenic and Nonpathogenic Hemoflagellates, Cryptobia spp., of Fishes

Acid phosphatase (ACP) was detected in whole-cell lysates, membrane-bound and water-soluble fractions of Cryptobia salmositica (pathogenic and nonpathogenic vaccine strains), Cryptobia bullocki, and Cryptobia catostomi using p-nitro-phenylphosphate as the substrate. High activities were in acidic pH (3.0-5.5) and the optimal pH was 5.0 Highest ACP activity was in the membrane-bound fraction. The pathogenic strain of C. salmositica had significantly higher total ACP activity than the vaccine strain and the other 2 species. However, the activity in the pathogenic C. salmositica decreased significantly with prolonged in vitro cultivation. The membrane-bound ACP of the pathogenic C. salmositica had highest resistance to the ACP inhibitor, sodium tartrate.

Induction of renal damage in rats by a diet deficient in antioxidants

Male albino rats. age 28 days, were fed a diet containing both vitamin E (10 g/kg) and selenium (5 mg/kg) or a diet lacking these antioxidants. Animals were examined for renal function after 4, 8, 12 and 16 wk on the respective diets. After 8 wk, animals on the deficient diet weighed less than controls (15%, p<0.01), and this became more pronounced by 16 weeks (25%, p<0.01). Expressed on a body weight basis, kidney wet weights did not differ between the two groups of animals. Urine volume increased in the animals fed the deficient diet at 8 weeks (66%, p<0.01) and this was maintained at 16 weeks (35%, p<0.01). Similar increases were observed for the rates of excretion of urinary total protein (77% elevation at 16 wk, p<0.01) and urinary acid phosphatase (51% elevation, p<0.01). At 16 wk, the specific activity of renal acid phosphatase in the animals given the deficient diet was reduced in cortex (57%, p<0.01) and medulla (20%, p<0.01), but not in papilla. These data indicate that dietary antioxidant deficiency causes progressive and pronounced renal damage.

Effect of tobacco consumption on the function of male accessory sex glands

The function of accessory sex glands in 29 tobacco smokers, 25 tobacco chewers and 30 non-users of tobacco was investigated by determining the ejaculate contents of various glandular markers: N-acetyl amino sugar and total phosphate (seminal vesicles) zinc and acid phosphatase (prostate gland), and alpha-1,4-glucosidase (epididymis). Both vesicular and prostatic parameters were reduced significantly in smokers compared with non-users of tobacco, whereas these parameters were unchanged in tobacco chewers. This difference may be due either to the difference in constitution of xenobiotics emitted as a result of burning tobacco, or to differences in the pharmacokinetics of the two forms of tobacco consumption. The activity of alpha-1,4-glucosidase was significantly lowered in both types of tobacco users. It is concluded that use of tobacco, especially by smoking, impairs the secretory function of accessory sex glands in man.

Prostatic acid phosphatase assay with self-indicating substrate 2,6-dichloro-4-acetylphenyl phosphate

We characterized six self-indicating substrates, synthesized as the derivative compounds of acetylphenyl phosphate, for serum prostatic acid phosphatase (PAP) activity. One of the substrates, 2,6-dichloro-4-acetylphenyl phosphate (DCAPP), is superior to others in terms of stability, affinity, and low Km for PAP. The hydrolyzed product, 2,6-dichloro-4-acetylphenol (DCAP), has a maximum absorption at 334.2 nm, a pKa of 4.15, and a molar absorptivity at 340 nm of 21,490 L.mol-1.cm-1 in citrate-HCl buffer, pH 5.4. PAP activity was assessed by subtracting tartaric acid-inhibited acid phosphatase activity from total acid phosphatase activity. Our assay system involving DCAPP is a unique kinetic method that shows good reproducibility, wide analytical dynamic range, and high specificity for PAP. Moreover, it is easily adaptable to automated analyzers because the product, DCAP, can be monitored at 340 nm.

A comparative study of tumor markers of adenocarcinoma prostate

The present study deals with the evaluation and comparison of the tumor markers for prostatic carcinoma—The Total Acid Phosphatase (ACP Total) and its Prostatic Fraction (ACP PF) estimated by the enzyme kinetic method, an immunoreactive Prostatic Acid Phosphatase (PAP) and Prostate Specific Antigen (PSA) estimated by enzyme immunoassay. The comparison of all four markers revealed that there was no perfect positive correlation between any of these four markers. PSA had shown a better correlation with ACP Total and its prostatic fraction ACP PF. No correlation was observed between PSA and PAP. Of the four markers PAP exhibited a very low sensitivity, positive and negative predictive values. PSA had shown an absolute specificity, sensitivity, positive and negative predictive values for adenocarcinoma prostate. PSA levels in all phases of disease showed a 100% correlation with disease status. Being a marker with very high tissue specificity and sensitivity, it is revolutionizing the diagnosis of prostatic carcinoma. Hence, it could be used effectively for screening of elderly people over 50 years of age who are at high risk for developing prostatic carcinoma for early diagnosis of this disease.