Prostatic Phosphatase Research Articles

Tumor markers in the process of radiotherapy of patients with prostate cancer

Purpose of the work – to assess the level of tumor markers in the blood of patients with prostate cancer (PC) in the course of their radiation therapy (RT). Material and methods. The study included 195 men with prostate cancer aged 52 to 82 years. The duration from the moment of diagnosis of the disease was 3 years on average. The Gleason score parameters averaged 6 points, the ratio of peripheral to central tumor form was 3: 1, adenocarcinoma was diagnosed in 94% of the cases, and giant cell carcinoma – in 6%. The average tumor stage parameter was 3 relative units, maturity was 3 points. Blood levels of testosterone (TS), luteinizing hormone (LT), insulin-like growth factor 1 (ISF), alkaline phosphatase (AP), acid prostatic phosphatase (APP) and glycosyl hydrolase (GH) activities were evaluated.Results. An increase in the level of tumor markers is observed in 50% - 100% of the patients, which depends on the expansion, size, localization, stage and degree of differentiation of the tumor process, the nature of its metastasis to the lymph nodes, distant organs and skeleton. Radiation therapy in patients with prostate cancer has an effect on integral changes in tumor markers, in particular, on the restoration of blood PSA parameters and APP activity; moreover, in the group of examined patients with an early initiation of RT after prostatectomy, positive changes in the studied parameters are more significant, do not depend on the expansion, size, and the degree of maturity of the neoplasm, but the nature of the dynamics of the levels of PSA, APP and ISF is closely related to the initial indicator of the severity of the tumor process course.Conclusion. The study of the level of tumor markers in the blood will have clinical and prognostic significance for assessing the efficacy of treatment of a tumor process, and contribute to the development of criteria for diagnosing complications of radiation therapy.

Clinicopathological features of metastatic castration-resistant prostate cancer

Objective To investigate the histopathological characteristics of metastatic castration-resistant prostate cancer. Methods Clinical findings, morphologic features, immunophenotypes of androgen receptor(AR), prostatic specific antigen(PSA), chromogranin A(CgA), and phosphatase and tensin homolog deleted on chromosome ten(PTEN)(by EnVision method)of 178 core biopsies from patients with metastatic castration-resistant prostate cancer encountered from January 2010 to January 2015 from West Coast Dream Team were analyzed. Results In the 178 cases, 51(28.7%)were diagnosed with typical adenocarcinoma, 41 cases(23.0%)with intermediate atypical carcinoma and 24 cases(13.5%)with small cell carcinoma.The expression of PSA was significantly lower in patients with intermediate atypical carcinoma or with small cell carcinoma(31/39, 79.5%)than in those with adenocarcinoma(20/20, 100%). The expression of PTEN was significantly higher in patient with intermediate atypical carcinoma or with small cell carcinoma(31/46, 67.4%)than in those with adenocarcinoma(19/41, 46.3%). Conclusions Three histopathological subtypes exists in metastatic castration-resistant prostate cancer, including adenocarcinoma, intermediate type atypical carcinoma and small cell carcinoma.They are entities with unique pathological features.The expression of PTEN is significantly lower in the patients with intermediate atypical carcinoma or with small cell carcinoma than in those with adenocarcinoma, indicating that intermediate atypical carcinoma and small cell carcinoma are different from typical adenocarcinoma in tumor occurrence and progression. Key words: Prostatic neoplasms; Neoplasm metastasis; Testosterone

Evaluation of the “Antidotal” Potential of Mangifera indica L. leaves extract on sodium arsenate exposed male Wistar rats using some biochemical markers

In order to evaluate the antidotal potential of Mangifera indica L leaves extract on sodium arsenate exposed male Wistar rats using some biochemical markers, forty-two apparently healthy male Wistar rats (weight range 120-160 g) were used in the study. The animals were randomly separated into six groups. Other than groups “A” (non-exposed control) and “B” (exposed control), groups; C, D, E, and F respectively were treated with different dosages of Mangifera indica L extract viz., l00 mg/kg and 200 mg/kg extract. Volumes of extract administered did not exceed 0.2 ml regardless of the body weight of the animal respectively. Some biochemical parameters assessed were: serum protein, albumin, conjugated bilirubin, unconjugated bilirubin (ICB), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP),gamma-glutamyl transferase (GGT), urea, creatinine, creatine kinase (CK), lactate dehydrogenase (LDH),acid phosphatase, prostatic phosphatase, serum lipid profile, that is total cholesterol, low density lipoprotein cholesterol (LDL), high density lipoprotein cholesterol (HDL), and the hormones, testosterone and luteinizing hormone levels. Group “B” had significantly (P Mangifera indica treated groups and to the non-exposed control. Testosterone and LH were significantly (P Mangifera indica treated groups and group “A”. This observation could be attributed to adverse effect of toxicosis on exposure to animals in group “B”. Antitodal property of the extract, due to one or more of its phytochemicals such as flavonoids, tannins alkaloid and anthraqunones could be the most probable reason for potential therapeutic potential. Conclusively, this observation gives credence to its cytoprotective and antitodal properties.

Effects of halofantrine, sulfadoxine/pyrimethamine and sulfametopyrazine/pyrimethamine on the concentration of serum phosphatases in male guinea pigs

Background: New single and combined antimalarial therapies developed as a result of resistant strains of Plasmodium falciparum raise new toxicological concerns and considerations. Aims: To establish and compare the dose- and time-dependent effects of three clinically used antimalarials agents on serum enzymes; alkaline phosphatase, prostatic phosphatases and acid phosphatase tartarate. Methods: Sixty male guinea pigs of average weight 500±5 gms were placed in twelve cages of five each. They were divided into four major groups, of three subsets each. The first group (control) received only water and food. The second, third and fourth groups of three cages of five animals each were given halofantrine hydrochloride (4.0-16.0 mg/kg), sulfadoxine/pyrimethamine (12.5-50.0 mg/kg) and sulfametopyrazine/pyrimethamine (12.5-50.0mg/kg) respectively in increasing doses of half clinical, clinical and twice clinical doses. In the time-dependent experiment, the above stated protocol was repeated at different time intervals with a set of five animals each. Blood was taken at 24 hrs, 7 and 28 days respectively for biochemical analysis. Analysis of variance at 0.05 level of significance was used to test differences between groups. Results: There were dose-dependent increases in alkaline phosphatases from14.7+0.58 to 38+2.0 IU/l (P = 0.046), 16.5+2.12 IU/l to 31.3+1.53 IU/l (p < 0.05) and 14.5+2.1 to 23+1.0 IU/l (P = 0.001) for sulfadoxine/pyrimethamine, halofantrine hydrochloride and sulfametopyrazine/pyrimethamine respectively. The time-dependent results show a progressive increase in these values maximizing on the seventh day, with a reversal (decreases) on the twenty eight day. Conclusion: These agents produced dose- and time-dependent effects on serum phosphatases. Keywords : Antimalarials, Serum phosphatases, Toxicity

The effects of mesterolone on the male accessory sex organs, on spermiogram, plasma testosterone and FSH.

42 subfertile male ambulatory patients were treated with Proviron. Moderate oligoastheno-teratozoospermia was the most common injury in sperm analysis. The treatment did not change the amount of plasma FSH, testosterone or prostate phosphatase. Acid phosphatase and citric acid of semen showed an increased activity with mesterolone treatment. The amount of fructose decreased, it is probably due to the increased number of spermatozoa, which need more fructose for their metabolism respectively. The sperm of 93% of the patients improved or stayed unchanged. 30% of the patients developed normozoospermia. 6 pregnancies were achieved.

Na-tartrate in the treatment of chronic nonbacterial prostatitis.

In an open investigation 20 patients with a history of chronic abacterial prostatitis (24 month duration) and of multiple therapies were treated by 0.07-0.15/kg Na-tartrate. Na-tartrate, a known inhibitor of the prostatic fraction of acid phosphatase in laboratory studies, was given orally for 12 weeks after dissolution in fluids and well tolerated. 13 patients completed the study and 3 spermiograms at 6 weeks interval were performed. 11/13 patients demonstrated increased sperm count (44.5 +/- 26.4 leads to 55.06 +/- 34.7) elevated seminal prostatic phosphatase (727 +/- 114.7 leads to 810.8 +/- 55.7) and improved seminal testosterone (2.86 +/- 1.6 leads to 4 +/- 1.4) while seminal y-GT was not significantly changed. Polyamine excretion showed a distinct elevation for putrescine 17.7 +/- 5.3 leads to 28.5 +/- 6.8. Clinically the 11 from 13 patients presented with marked improvement in a 4 months follow-up.

Mature cystic teratoma of the ovary with male accessory sexual glands including seminal vesicles, prostatic tissue, and bulbo-urethral glands: a case report

We present an extremely rare case of a benign cystic ovarian teratoma with structures of male accessory sexual glands. The patient was a 30-year-old woman. A unilocular cystic tumor, measuring 5 cm in the largest diameter, was found in her right ovary and was removed. The teratoma contained epidermis, skin appendages, respiratory and intestinal epithelia, cartilage, muscle, and nervous and connective tissue. In addition to these histologically mature tissues, there were nodules with prostatic acini, prostate duct-like structures strongly positive for prostate-specific antigen and acid prostatic phosphatase, structures resembling Cowper's glands, and seminal vesicles surrounded by fibromuscular stroma. To our knowledge, this is the first case in the English literature describing seminal vesicles associated with prostatic tissue and bulbo-urethral glands in a mature ovarian teratoma.

Kinetic intermediates of unfolding of dimeric prostatic phosphatase.

Kinetics of guanidine hydrochloride (GdnHCl)-induced unfolding of human prostatic acid phosphatase (hPAP), a homodimer of 50 kDa subunit molecular mass was investigated with enzyme activity measurements, capacity for binding an external hydrophobic probe, 1-anilinonaphtalene-8-sulfonate (ANS), accessibility of thiols to reaction with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and 2-(4'-maleimidylanilino)naphthalene-6-sulfonate (MIANS) and ability to bind Congo red dye. Kinetic analysis was performed to describe a possible mechanism of hPAP unfolding and dissociation that leads to generation of an inactive monomeric intermediate that resembles, in solution of 1.25 M GdnHCl pH 7.5, at 20 degrees C, in equilibrium, a molten globule state. The reaction of hPAP inactivation in 1.25 M GdnHCl followed first order kinetics with the reaction rate constant 0.0715 +/- 0.0024 min(-1) . The rate constants of similar range were found for the pseudo-first-order reactions of ANS and Congo red binding: 0.0366 +/- 0.0018 min(-1) and 0.0409 +/- 0.0052 min(-1), respectively. Free thiol groups, inaccessible in the native protein, were gradually becoming, with the progress of unfolding, exposed for the reactions with DTNB and MIANS, with the pseudo-first-order reaction rate constants 0.327 +/- 0.014 min(-1) and 0.216 +/- 0.010 min(-1), respectively. The data indicated that in the course of hPAP denaturation exposure of thiol groups to reagents took place faster than the enzyme inactivation and exposure of the protein hydrophobic surface. This suggested the existence of a catalytically active, partially unfolded, but probably dimeric kinetic intermediate in the process of hPAP unfolding. On the other hand, the protein inactivation was accompanied by exposure of a hydrophobic, ANS-binding surface, and with an increased capacity to bind Congo red. Together with previous studies these results suggest that the stability of the catalytically active conformation of the enzyme depends mainly on the dimeric structure of the native hPAP.

Continuous monitoring of prostatic acid phosphatase using self-indicating substrates

Background: The continuous measurement of acid phosphatase (EC 3.1.3.2) activity in serum represents an analytical task not yet sufficiently accomplished. Methods: Introducing two novel substrates—2-chloro-4-nitrophenyl phosphate (CNP-P), which was preferred, and 4-nitronaphthyl-1-phosphate (NN-P)—an alternative assay to measure enzymatic activity was developed and compared with a modification of Hillmann's method (azo coupling of released naphth-1-ol with a diazonium compound). Apart from different substrate concentrations of 2-chloro-4-nitrophenyl phosphate, 4 mmol/l, and naphthyl-1-phosphate (N-P), 8 mmol/l (with Fast Red TR, 5 mmol/l), respectively, following identical conditions were selected: Citrate, 50 mmol/l, pH 5.75; pentane-1,5-diol, 150 mmol/l; tartrate, 60 mmol/l; 37 °C. Results: Whereas intensity and stability of the azo dye unpredictably depend on the albumin concentration of the sample, the direct test with 2-chloro-4-nitrophenyl phosphate resisted sample interferences, showed no intrinsic hydrolysis by albumin, relied on stable reagents and proved superior in sensitivity, precision and ease of handling. In measuring prostatic phosphatase, the proposed procedure closely correlated with Hillmann's method. The preliminary 0.95-reference intervals for adults were 1.2–3.9 kU/l and 5.8–14.8 U/l for total activity, respectively. Conclusions: The direct assay of the enzyme is suited as an economic, rapid and robust method for mechanized or manual use.

Effect of tartaric acid on conformation and stability of human prostatic phosphatase: an infrared spectroscopic and calorimetric study.

The solution structure and thermal stability of human prostatic acid phosphatase (hPAP) in the absence and in the presence of tartaric acid were studied by Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The temperature dependence of the infrared spectrum and DSC scans indicate that hPAP undergoes thermal unfolding at a temperature between 49.5 and 52.5 degrees C. Binding of tartaric acid does not lead to major changes in the secondary structure of hPAP, however, hPAP with bound tartaric acid shows a significantly increased thermal stability. These results helped to better understand the mechanism of hPAP unfolding at the elevated temperature.

Cooperative kinetics of human prostatic acid phosphatase

The steady-state kinetics of hydrolysis reaction catalysed by human prostatic acid phosphatase (PAP) by using 1-naphthyl phosphate, phenyl phosphate and phosphotyrosine as substrates has been studied at pH 5.5. The substrate binding curves were sigmoidal and Hill cooperation coefficient h was higher than 1 for each of the examined compounds. Thus, human prostatic acid phosphatase kinetics exhibits positive cooperativity towards the studied substrates. The extent of cooperativity was found to depend on the substrate used and on enzyme concentration. The highest cooperativity of PAP was observed for 1-naphthyl phosphate and the lowest for phosphotyrosine. When prostatic phosphatase concentration increased, Hill cooperation coefficient ( h) and half saturation constant ( K 0.5) both grew, but the catalytic constant ( k cat) remained constant, for each of the substrates studied. Ligand-induced association–dissociation equilibrium of the active oligomeric species (monomer-dimer-tetramer-oligomers) is suggested.

Molten globule as an intermediate on the human prostatic phosphatase folding pathway.

Human prostatic acid phosphatase (hPAP, EC.3.1.3.2), a secretory homodimeric protein was denatured in 6 M urea, pH 2.5, and refolded by dilution at pH 7.2 with recovery of the enzymatic activity and dimeric structure. Circular dichroism, intrinsic fluorescence and chromatographic analysis of renaturating protein suggested that the kinetic intermediate of the hPAP folding is a monomer which displays a molten globule state (R. Kuciel, A. Mazurkiewicz & W.S. Ostrowski, 1996, Int. J. Biol. Macromol. 18, 167-175). To confirm these data experiments were performed to estimate the interaction of the renaturating protein with dyes and amphipathic lipid structures. Increased binding of the hydrophobic probe 1-anilinonaphthalene-8-sulfonate and Congo Red to the refolding enzyme supported the existence of molten globule state with the relaxed beta-structure in the renaturating protein. Presence of liposomes, included in the renaturation mixture as a model of acid phospholipid, resulted in perturbations of the human PAP refolding process. Some folding intermediates were bound to phosphatidylserine liposomes or, alternatively, water soluble, inactive aggregates were formed.

Prostate-specific antigen (protein and mRNA) analysis in the differential diagnosis and staging of prostate cancer

We analyzed complexed and free prostate-specific antigen (PSA), the free/total PSA and complexed/free PSA ratios, acid phosphatase, and prostatic phosphatase in serum from 36 patients with prostatic carcinoma and from 48 non-neoplastic control patients (20 with prostatitis and 28 with benign prostatic hyperplasia). Receiver-operating characteristic plots showed that serum PSA was the most efficient variable, singly used, in discriminating neoplastic from non-neoplastic patients. At a cut-off value of 10.0 ng/ml, serum PSA had a diagnostic sensitivity of 87% and a diagnostic specificity of 83%. In particular, three patients with prostatic carcinoma and twenty non-neoplastic controls had serum PSA levels of between 4 and 10 ng/ml. The subsequent analysis of the serum free/total PSA ratio, in this subgroup, using a cut-off level of 15%, allowed us to classify correctly all prostatic cancer cases and 18 20 non-neoplastic diseases. We next analyzed PSA mRNA in circulating cells using an improved reverse-transcriptase polymerase chain reaction dot blot procedure, from six patients with prostatic carcinoma with distant metastases, and in seventeen with localized cancer. The analysis had a high sensitivity (up to dilutions 1:10 6 of total RNA from prostatic cancer cells vs total RNA from normal blood cells). The analysis revealed circulating micrometastatic cells in 3 6 (50%) cases of metastatic cancer and in 4 17 cases of localized cancer. To conclude, serum totalcombined with the free/total PSA ratio is a very efficient algorithm in discriminating neoplastic from non-neoplastic prostatic diseases, while other mRNA species must be analyzed, in addition to PSA mRNA, in circulating cells to increase the efficiency in detecting metastatic prostatic cancer.

Biochemical studies of the effect of gossypol consumption on cholesterol and residual glucose in fed and protein-energy deficient rats

Some biochemical parameters were investigated on gossypol consumption, and were correlated to the level of cholesterol and residual glucose in rat liver. Two groups of animals were used, one group was fed with normal protein diet and the other set was fed with low protein diet. The results show that gossypol produced no apparent biochemical aberration in the liver of normal protein fed and low protein fed rats. It also had no effect on glucose-6-phosphatase and prostate phosphatase. Gossypol consumption had a significant effect on alcohol dehydrogenase. These results indicate no direct involvement of gossypol in sugar uptake but profound influence on the regulation of cholesterol level in the liver.

Human prostatic acid phosphatase: Selected properties and practical applications

Human prostatic acid phosphatase (EC 3.1.3.2) is a non-specific phosphomonoesterase, synthetized and secreted into seminal plasma under androgenic control. The enzyme is a dimer of molecular weight around 100 kDa. Gene coding this protein is localized on chromosome 3. Since many years prostatic phosphatase has been used as a marker for diagnosis and therapy control of cancer of the prostate gland. The biological role of this enzyme, however, remains unknown and needs further exploration.

Heterologous expression of human prostatic acid phosphatase and site-directed mutagenesis of the enzyme active site.

Earlier covalent modification experiments indicated that histidine, arginine, and carboxylic acid residues were involved in the catalytic mechanism of the acid phosphatase enzyme from human prostate. The present study utilizes site-directed mutagenesis to evaluate the catalytic importance of 7 residues of human prostatic phosphatase, namely His12, His257, Asp258, Arg11, Arg15, Arg54, and Arg79, which are highly conserved in the sequences of high molecular weight acid phosphatases. Both wild type and mutant enzymes were expressed in the yeast Saccharomyces cerevisiae as fusions to a leader peptide of prepro-alpha-factor, which resulted in the secretion of processed proteins into the culture media. The kinetic parameters of the recombinant wild type enzyme were shown to be effectively identical with those of the native protein. The circular dichroism spectra of wild type and mutant proteins were effectively superimposable. Replacement of His12, which was previously shown to be transiently phosphorylated during catalysis, as well as the adjacent Arg11, both resulted in > 10(4) reductions of enzyme activity. Consistent with a postulated involvement of arginine residues in substrate binding, large reductions in Vmax caused by the R15A and R79A mutations were accompanied by marked increases in Km. However, the mutation R54A had only small effects on kinetic parameters. Essential roles for both His257 and Asp258 were also established. The D258A mutation not only caused larger decreases of Vmax toward p-nitrophenyl phosphate compared to the effect of the H257A mutation, but it also caused the reaction to become more sensitive to the nature of the leaving group. It is concluded that His12 and Asp258 are involved in the formation of the phosphoenzyme intermediate as an acceptor of the phospho group and a proton donor for the substrate leaving group, respectively, while His257 may participate in substrate binding or may facilitate the breakdown of the phosphoenzyme by maintaining the protonated state of the phosphorylated His12 residue.

Lead toxicity on the accessory reproductive organs in humans

The prostatic function of 50 males was studied. Thirty of the males were exposed to lead and 20 males were not exposed. The exposed individuals included workers at a battery manufacturing plant. The analysed variables were both acid and prostatic phosphatases. The level of exposure was estimated by means of lead concentration in blood. The total acid phosphatase activity was significantly higher in exposed subjects, when compared to control subjects. This was not the case with the acid prostatic phosphatase. These findings suggest that an extensive exposure to lead initially causes a direct prostatic toxicity likely to be reflected in alteration of seminal characteristics.

Experience with prostate-specific antigen in prostatic carcinoma

A total of 71 prostatic tumour patients and 45 prostatic adenoma patients were tested for prostate-specific antigen (PSA), immunological prostatic acid phosphatase (PAP) concentration as well as serum prostatic phosphatase (SPP) and enzymic serum phosphatase. It was found among untreated patients that PSA showed the highest percentage of pathologic affection in each stage. PSA, on the evidence of clearance test in the initial days of therapy and after a follow-up period of several months, gave a good picture of the course that the disease had taken.

Adenosine metabolizing enzymes in seminal plasma of bull and man: A comparative study

1. 1. Adenosine metabolizing enzymes in seminal plasma of man and bull have been investigated. 2. 2. A different level of 5′-nucleotidase activity has been found in two seminal plasmas: in bull 5′-nucleotidase represents 80% of the total AMP dephosphorylating enzymes while in man 5′-nucleotidase represents only 1.3% of the total AMP dephosphorylating activities. 3. 3. Apart from the different levels of 5′-nucleotidase activity, different kinetic parameters have been reported for 5′-nucleotidase, acid prostatic phosphatases, ADA and PNP. 4. 4. Adenosine kinase, xanthine oxidase and AdoHcy-hydrolase have not been detected in the seminal plasma of man and bull.

Vanadate monomers and dimers both inhibit the human prostatic acid phosphatase

A combination of enzyme kinetics and 51V NMR spectroscopy was used to identify the species of vanadate that inhibits acid phosphatases. Monomeric vanadate was shown to inhibit wheat germ and potato acid phosphatases. At pH 5.5, the vanadate dimer inhibits the human prostatic acid phosphatase whereas at pH 7.0 it is the vanadate monomer that inhibits this enzyme. The pH-dependent shift in the affinity of the prostatic phosphatase for vanadate is presumably due to deprotonation of an amino acid side chain in or near the binding site resulting in a conformational change in the protein. pH may be a subtle effector of the insulin-like vanadate activity in biological systems and may explain some of the differences in selectivity observed with the protein phosphatases.