Determination Of Acid Phosphatase Activity Research Articles

Responses of Soil Phosphorus Fractions to Land-Use Change in Colombian Amazon

Intensive land-use change, the overgrazing of pastures, and the poor soil management in the Amazon region induce significant soil chemical degradation, causing alterations in the soil phosphorus (P) dynamics. Here, we studied the changes in P fractions and availability throughout the soil profile along a chronosequence composed of four study areas representing the typical land-use transition from forest to pasture for extensive cattle ranching in the Colombian Amazon region: (i) Forest—Deforested—Pasture 4 years old and Pasture established >25 years after deforestation. Soil samples collected at 0–10, 10–20, 20–30, and 30–40 cm depth were used for the sequential fractionation of P, determination of acid phosphatase activity and soil organic carbon (C) content, and calculation of C:organic P (Po) ratio and P stocks. Our results showed that the land-use change caused a decrease of 31.1% in the fractions of labile inorganic P, with the mineralization of organic P by phosphatase enzyme playing an essential role in the P availability. Although according to the C:Po ratio of the deeper layer the P seems to be sufficient to satisfy the plant needs of all the land uses assessed, the exploitation of soil nutrients in pastures reduced by 6.1% the moderately and non-labile P stock. Given the role of cattle ranching in the economy of tropical countries, it is imperative to adopt strategies of soil P management to improve P-use efficiency, avoiding the degradation of grazing land resources while ensuring the long-term sustainability of rangeland livestock and decrease further deforestation of the Amazon rainforest.

Colorimetric determination of acid phosphatase activity and inhibitor screening based on in situ polymerization of aniline catalyzed by gold nanoparticles.

A colorimetric assay for acid phosphatase (ACP) was constructed that isbased on in situ polymerization of aniline catalyzed by gold nanoparticles (AuNPs). Aniline can be polymerized by ammonium persulfate (APS) in acidic condition and form gold-polyaniline core-shell nanoparticles (Au@PANI NPs) in the presence of AuNPs with the assistance of sodium dodecyl sulfate (SDS). AuNPs were also found to accelerate the polymerization process of aniline and thus shorten the reaction time. Upon the introduction of ascorbic acid (AA), the oxidant APS was consumed via the redox reaction. That led to the suppression of the formation of PANI. Consequently, ACP activity can be supervised on the basis of hydrolysis of 2-phospho-L-ascorbic acid trisodium salt (AAP) catalyzed by ACP to release AA. With the increase of ACP activity, the intensity ratio of the absorbance at λ705 nm (A705) and the absorbance at λ530 nm (A530) gradually decreased and the color gradually changed from dark-green to light-green to blue-gray to purple and eventually to pink. This method for ACP determination worked in the range 0.40 to 2.00U·L-1. The detection limit is 0.043U·L-1. The assay was applied to determine ACP in human serum. The recovery ranged from 81.0 to 104.6%. Relative standard deviation was less than 5%. This suits the request forbiological sample analysis. Graphical abstract Schematic presentation of the colorimetric determination of acid phosphatase activity and inhibitor screening based on in situ polymerization of aniline catalyzed by gold nanoparticles. : acid phosphatase (ACP); : gold nanoparticles (AuNPs); : gold-polyaniline core-shell nanoparticles (Au@PANI NPs); ascorbic acid (AA); 2-phospho-L-ascorbic acid trisodium salt (AAP).

Determination of Acid Phosphatase Activity in Nervous Tissue

High levels of acid phosphatase activity in nerve cells allow methods for its detection to be used in morphofunctional studies of nervous tissue elements. Using neurons in the stellate ganglion in cows as an example, the bodies and processes of nerve cells are shown to have different levels of enzyme activity. Nerve endings have consistently high levels. This method provides for detailed morphological investigation of neurons and assessment of their interactions. This report provides a detailed description of an improved method for identifying acid phosphatase activity in nervous tissue structures.

Study of the Effect of Zinc on the Fertility and Acid Phosphatase Activity and Fructose Concentration in Male Swiss Albino Mice ( Mus Musculus)

This study was conducted to elucidate the effect of different concentrations of zinc element on male fertility of swiss albino mice by using atomic absorption, and determination of acid phosphatase activity ( Ec.3.1.3.2) compared with control groups. The mice were divided into three groups ( 10 mice / group ) and they were three months old. The first group was treated with 0.5 mg / kg body weight and the second group with 1.5 mg / kg body weight with zinc, while the third group considered as control group. The mice treatment was for 15 and 30 days for the two experiment groups. The results showed a significant increase ( p≤0.05 ) in weight of testes in the group which treated by concentration 1.5 mg / kg b.w compared with control group. Significant increase (p≤0.05) was also found in testes weight when using the high concentration of zinc compared with the low concentration. Results also showed a significant increase in sperm count of groups treated with zinc as compared with control group. Significant increase (p≤o.o5) in sperm motility was detected in both the first and second groups as compared with control . Significant decrease (p≤0.05) in slugish sperm in group with high concentration of zinc as compared with control. Significant increase in both groups in acid phosphatase activity and the concentration of fructose treated with concentrations 0.5 and 1.5 mg / kg b.w as compared with control.

Preliminary investigations on the standardization and quality control for the determination of acid phosphatase activity in seminal plasma

Background The determination of ACP in seminal plasma was considered as an appropriate biochemical marker to evaluate prostate function, as recommended by the WHO manual. However, few reports on the standardization and quality control for the determination of biochemical markers in seminal plasma have been documented. Methods Two frozen samples of seminal plasma with or without phenylmethylsulfonyl fluoride were determined for their acid phosphatase (ACP) levels. The ACP level and sperm concentration of each of 72 samples of seminal plasma obtained at 1000× g for 10 min or 3000× g for 15 min centrifugation were assayed. ACP activity in 10 samples of seminal plasma was measured immediately or standing for 30 min after dilution. The ACP levels in seminal plasma with or without chymotrypsin were also assayed. Results There was no significant difference of ACP levels ( P = 0.166) but of sperm concentrations ( P = 0.000) in seminal plasma obtained by centrifugation at different velocity. ACP activities in seminal plasma measured when standing for 30 min after dilution were significantly lower than those measured immediately after dilution ( P = 0.001). Both chymotrypsin and freezing–thawing had no apparent effect on the determination of ACP in seminal plasma. Conclusion The results indicated that standing time after dilution and centrifugation velocity should be standardized, and frozen seminal plasma could serve as the quality control products for the determination of ACP activity among different laboratories.

Electroanalytical determination of acid phosphatase activity by monitoring p-nitrophenol

The development of a modified electrode for the selective determination of p-nitrophenol (PNP) to be applied in the determination of the enzymatic activity (EA) of acid phosphatase using p-nitrophenylphosphate (PNPP) as substrate is described. Elimination of the interference of PNPP was made by covering the electrode surface using 25 μl of a 5% (m/v) Nafion ® solution. In comparison to the glassy carbon (GC) electrode without modification, the presence of the membrane promoted a reduction in the sensitivity and an increase in the detection limit for the determination of PNP, in addition to a decrease of the electron transfer constant. The GC electrode covered with a Nafion ® membrane showed a linear response range between 20 and 230 μmol l −1 for PNP, adjusted by the equation I P=0.157+0.0089 [PNP], for N=22 with a correlation coefficient of 0.9984. This electrode was used for the determination of the EA of acid phosphatase from castor bean seeds and an activity of 38.40 U mg −1 was found.

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.

Determination of acid phosphatase activity in heat treated milks by the Fluorophos Test System

The Fluorophos Test System® for measurement of acid phosphatase in fruit juices was used to determine the survival of acid phosphatase in heat treated cows' milk. The method was found to be applicable to milk, with reproducibility improved by use of a vortex mixer. It was sufficiently sensitive to provide a quantitative measure of the activity surviving heat treatments from 75 to 88° C for 15 seconds and would thus provide a rapid method for checking more severe heat treatments_ for instance, those used to enhance the destruction of Mycobacterium paratuberculosis.

A rapid method for determination of acid phosphatase activity of whole yeast cells

A simple and rapid procedure for determination of intracellular acid phosphatase activity without the need for disruption of cells is described. Candida lipolytica cell suspension was treated with 0.1% Triton X-100 for 30 min at room temperature and with intermittent shaking. The enzyme assay is carried out directly with the permeabilized cell suspension. Permeabilization of the yeast cells to p-nitrophenylphosphate by Triton X-100 provides almost 100% efficiency in determining the total acid phosphatase activity compared to results obtained with disrupted yeast cells

Cytochemical determination of acid phosphatase activity in isolated rat hepatocytes during starvation-induced proteolysis.

Acid phosphatase activity in isolated rat liver parenchymal cells has been investigated with quantitative histochemical means during short-term starvation, which leads to a considerable loss in protein mass in the parenchyma. Animals trained to a meal-feeding regime in which food was available during 1 h only per 24 h (using an automatic food dispensing machine), were sacrificed 6, 12, 18, 24 and 36 h after food had been withheld at the time point (23.00 h) of meal feeding. Acid phosphatase activity was analysed cytophotometrically in isolated hepatocytes incorporated into polyacrylamide gels before the enzyme reaction technique with the post-azo coupling was carried out. No indication could be found for any significant changes in the amount of acid phosphatase activity per individual hepatocyte during the entire period of fasting, as compared with two time points (11.00 and 23.00 h) before the theoretical onset of fasting. It is concluded that the considerable enhancement of protein degradation in the lysosomal apparatus during fasting is not reflected by changes in the cellular acid phosphatase activity.

Interferences in the direct kinetic determination of acid phosphatase activity.

Journal Article Interferences in the direct kinetic determination of acid phosphatase activity. Get access C W Small, C W Small Search for other works by this author on: Oxford Academic Google Scholar P McNutt P McNutt Search for other works by this author on: Oxford Academic Google Scholar Clinical Chemistry, Volume 30, Issue 4, 1 April 1984, Pages 594–595, https://doi.org/10.1093/clinchem/30.4.594 Published: 01 April 1984

Quantitative evidence of a silent gene P° of human red cell acid phosphatase in south Polish population

The distribution of red cell acid phosphatase types in 3244 unrelated Polish adults is reported. Gene frequencies P a = 0.3594, P b = 0.5643 and P c = 0.0763 were obtained. In a forensic case of disputed paternity an apparent mother/child incompatibility respect to red cell acid phosphatase was found, the mother appearing as type B and the child as type A. Determination of acid phosphatase activity suggested the presence of a silent gene P°. The phosphatase levels were about half the values expected, as determined in 237 adults representing the different phenotypes.

An automated continuous-monitoring procedure for the determination of acid phosphatase activity in serum

The method of Hillmann, in which hydrolysis of α-naphthyl phosphate by acid phosphatase is coupled to the formation of an α-naphthol-Fast Red TR azo-compound, has been adapted for use with the LKB Produkter AB 8600 reaction rate analyzer. Factors which affect the reproducibility of the method are described and its performance is shown to be superior to that of a manual phenyl-phosphate procedure.

Quantitative determination of acid phosphatase activity detected in tissue sections according to the BURSTONE method

Quantitative determination of the activity of acid phosphatase detected in tissue sections using Burstone's method. The experiment was performed on inbred rats. A reaction for AcP according to Burstone was made in liver sections, incubation time was fixed experimentally. The material obtained from each animal was divided into 3 parts. One of them was used for enzymatic estimations, the 2nd for planimetry of sections, and the 3rd was eluted by N,N-dimethylformanid, and the obtained coloured product was subjected to spectrophotometric analysis at the Spectrophotometer Unicam SP 8000 B. Homogeneity of the coloured product eluted from the sections as well as its identity with the coloured product obtained during the reaction with a preparation of crystalline acid phosphatase was found. Repeatability of the obtained activity for the inbred animals was shown. The obtained results were subjected to statistical analysis. The method of quantitative estimation of AcP activity may be useful in histoenzymatic works.

Release of lysosomal enzymes during post mortem conditioning and their relationship to tenderness

SummarySamples of the M. longissimus dorsi were taken from beef carcasses 1 hr, 24 hr, 5 days, 7 days and 14 days post mortem. Tenderness (shear force), the amount of protein and hydroxyproline in filtered muscle homogenates, the activity of β‐glucuronidase in muscle fractions and the electron histochemical determination of acid phosphatase were measured at these post mortem times.Shear force was a maximum at 24 hr and decreased thereafter to a value less than that of the 1 hr samples by 14 days. The amount of protein and hydroxyproline in filtered muscle homogenates was a minimum at 24 hr and increased to a maximum at 14 days. The specific activity of free β‐glucuronidase (a lysosomal enzyme) increased with post mortem ageing, while the specific activity of bound (sedimentable) β‐glucuronidase decreased with post mortem ageing, suggesting that this enzyme is released during the conditioning period. Electron histochemical determination of acid phosphatase activity showed the activity to be localized in specific areas around the I‐band at early post mortem times: the activity was more dispersed throughout the cell at later post mortem times.

Preparation of histochemical sections for quantitative determination of acid phosphatase activity by means of laser microanalysis.

Based on former experiments dealing with a quantitative assay of acid phosphatase (acP) activity in histological sections by means of a laser microanalysis. The authors present results of investigations concerning optimal conditions needed for this purpose. The investigations were performed on rat liver sections. The realtion between incubation time and depth of acP reaction appearing in tissue blocks was investigated along with the dependence of photometric readings on thickness of histological sections and on incubation times. From results of experiments it appears that optimal conditions for quantitative determination of acP activity using the proposed laser method are provided by sections 30 to 50 μ in thickness which have been incubated in the substrate containing medium for 45 min. Sections thinner than 30 μ are not recommended for quantitative assay with this method because of low accuracy of readings caused by too low amounts of reaction product present there.

The turtle bladder. I. Morphological studies under varying conditions of fixation

The turtle bladder can be maintained in plastic chambers for prolonged periods and is currently being utilized to study ion transport. Its mucosa consists of two principal kinds of cell: a superficial cell fronting the lumen and containing many mucin granules and a basal cell distinguished by a relatively high nuclear-to-cytoplasmic ratio. Uncommonly a third cell type was observed characterized be rarefied cytoplasm, a flask shape, and small apical granules. The form of these cells varies with the degree of bladder stretch. Except for some alteration of cell organelle and sap density that may partially be due to fixation factors, the mucosa of in vitro preparations maintained as long as 7 hours resembles tissue immediately fixed. On the other hand, serosal cells under similar conditions show marked changes, thus eliminating them as an important factor in active ionic transport. Adding to their value as an investigative tool, histochemical techniques for the determination of acid phosphatase activity using light and electron microscopy can be applied to the stretched turtle bladder without the necessity of frozen sections. Unlike the toad bladder, which also is capable of in vitro ionic transport, the turtle bladder can function anaerobically and secrete hydrogen ions. Structurally both bladders are similar, but the turtle mucosal epithelium is distinguished by its greater content of endoplasmic reticulum, mitochondria, and mucin granules. It is hoped that in future studies these distinguishing anatomical and physiological characteristics can be related.

Automated Determination of Acid Phosphatase

Abstract A procedure is presented for the automated determination of acid phosphatase activity in biological materials using the Robot Chemist. Although either phenylphosphate and α-naphthylphosphate may be used as substrate in this analysis, the procedure is described in detail for serum acid phosphatase using α-naphthylphosphate in 0.1 M citrate, pH 5.2, since this substrate is more selective for prostatic acid phosphatase in human serum. Enzymically generated aα- naphthol is determined by the Emerson reaction (alkaline aminoantipyrine and ferricyanide), modified for use with this automated system. Correlations are presented between the results obtained on the Robot Chemist and the identical procedure developed for the AutoAnalyzer.

Automated Determination of Acid Phosphatase

Abstract A procedure is presented for the automated determination of acid phosphatase activity in biological materials from either plant or animal origin. The Technicon N-7 flow diagram has been simplified, and the reagents for the measurement of enzymatically produced phenolic substances have been modified without loss of range or sensitivity. Both phenylphosphate and α-naphthylphosphate, introduced by Babson et al. (2, 3) for serum acid phosphatase, may be used as substrates. The Emerson reaction (alkaline aminoantipyrine and ferricyanide) with α-naphthol forms a more stable and reproducible color than the coupled product with tetrazotized o-dianisidine (Babson). Supporting data for these modifications are included.