Sample records for serum alkaline phosphatase

  1. Low serum alkaline phosphatase activity in Wilson's disease.

    Shaver, W A; Bhatt, H; Combes, B


    Low values for serum alkaline phosphatase activity were observed early in the course of two patients with Wilson's disease presenting with the combination of severe liver disease and Coombs' negative acute hemolytic anemia. A review of other cases of Wilson's disease revealed that 11 of 12 patients presenting with hemolytic anemia had values for serum alkaline phosphatase less than their respective sex- and age-adjusted mean values; in eight, serum alkaline phosphatase activity was less than the lower value for the normal range of the test. Low values for serum alkaline phosphatase were much less common in Wilson's disease patients with more chronic forms of presentation. Copper added in high concentration to serum in vitro did not have an important effect on serum alkaline phosphatase activity. The mechanism responsible for the decrease in serum alkaline phosphatase activity in patients is uncertain.

  2. Bone mineralisation in premature infants cannot be predicted from serum alkaline phosphatase or serum phosphate

    Faerk, J; Peitersen, Birgit; Petersen, S


    BACKGROUND: The bone mineral content of premature infants at term is lower than in mature infants at the same postconceptional age. Serum alkaline phosphatase and serum phosphate are often used as indicators of bone mineralisation. OBJECTIVE: To analyse the association between bone mineral content...... and serum alkaline phosphatase and serum phosphate. METHODS: Serum alkaline phosphatase and phosphate were measured at weekly intervals during admission in 108 premature infants of gestational age below 32 weeks (mean (SD) gestational age 29 (2) weeks; mean (SD) birth weight 1129 (279) g). Bone mineral...... content was measured at term (mean gestational age 41 weeks) by dual energy x ray absorptiometry and corrected for body size. RESULTS: Serum alkaline phosphatase was significantly negatively associated with serum phosphate (p serum alkaline...




    ABSTRACT Objective: To study the relationship between the pre and post chemotherapy (CT) serum levels of alkaline phosphatase (AP) and lactate dehydrogenase (LDH), and the percentage of tumor necrosis (TN) found in specimens after the pre surgical CT in patients with osteosarcoma. Methods: Series of cases with retrospective evaluation of patients diagnosed with osteosarcoma. Participants were divided into two groups according to serum values of both enzymes. The values of AP and LDH were obtained before and after preoperative CT. The percentage of tumor necrosis (TN) of surgical specimens of each patient was also included. Results: One hundred and thirty seven medical records were included from 1990 to 2013. Both the AP as LDH decreased in the patients studied, being the higher in pre CT than post CT. The average LHD decrease was 795.12U/L and AP decrease was 437.40 U/L. The average TN was 34.10 %. There was no statistically significant correlation between the serums values and the percentage of tumoral necrosis. Conclusion: The serum levels values of AP and LDH are not good predictors for the chemotherapy-induced necrosis in patients with osteosarcoma. Level of Evidence IV, Case Series. PMID:27217815

  4. Cinacalcet Lowers Serum Alkaline Phosphatase in Maintenance Hemodialysis Patients

    Belozeroff, Vasily; Goodman, William G.; Ren, Lulu; Kalantar-Zadeh, Kamyar


    Background and objectives: Studies suggest an association between elevated serum alkaline phosphatase (AP) and increased mortality in hemodialysis patients, but the effect of existing therapies on AP is not fully understood. We assessed the effects of cinacalcet on AP in a secondary analysis of controlled trial data. Design, setting, participants, & measurements: This was a post hoc analysis of data from three 26-wk randomized, double-blind, placebo-controlled, phase 3 trials and a 26-wk double-blind, placebo-controlled extension trial that investigated cinacalcet in secondary hyperparathyroidism treatment in dialysis patients. Hemodialysis patients (n = 890) with intact parathyroid hormone ≥300 pg/ml and serum calcium ≥8.4 mg/dl received cinacalcet plus standard therapy or standard therapy alone for up to 52 wk. Total, not bone-specific, AP was assessed (proportion of cinacalcet/control subjects achieving a ≥20% or any AP reduction from baseline; the proportion of subjects with AP ≥120 U/L) at baseline; the end of titration; and study weeks 26, 42, and 52. Results: At 52 wk, a greater proportion of cinacalcet-treated patients had either a ≥20% (39 versus 18%) or any (58 versus 36%) AP reduction compared with control subjects, respectively. The likelihood of achieving either a ≥20% or any AP reduction (determined by relative proportion) was 2.33 (95% confidence interval 1.50 to 3.61) and 1.74 (95% confidence interval 1.31 to 2.31), respectively, at week 52. Cinacalcet treatment tended toward a decreased percentage of patients with AP ≥120 U/L (baseline, 42.6%; week 52, 30.6%) compared with control (35.0 to 48.6%, respectively). Conclusions: In this combined analysis of controlled trials of patients who were receiving hemodialysis, cinacalcet lowered total serum AP. PMID:19261825

  5. Associations between renal hyperfiltration and serum alkaline phosphatase.

    Se Won Oh

    Full Text Available Renal hyperfiltration, which is associated with renal injury, occurs in diabetic or obese individuals. Serum alkaline phosphatase (ALP level is also elevated in patients with diabetes (DM or metabolic syndrome (MS, and increased urinary excretion of ALP has been demonstrated in patients who have hyperfiltration and tubular damage. However, little was investigated about the association between hyperfiltration and serum ALP level. A retrospective observational study of the 21,308 adults in the Korea National Health and Nutrition Examination Survey IV-V databases (2008-2011 was performed. Renal hyperfiltration was defined as exceeding the age- and sex-specific 97.5th percentile. We divided participants into 4 groups according to their estimated glomerular filtration rate (eGFR: >120, 90-119, 60-89, and 120 mL/min/1.73 m2 showed the highest risk for MS, in the highest ALP quartiles (3.848, 95% CI, 1.876-7.892, compared to the lowest quartile. Similarly, the highest risk for DM, in the highest ALP quartiles, was observed in participants with eGFR >120 ml/min/1.73 m2 (2.166, 95% CI, 1.084-4.329. ALP quartiles were significantly associated with albuminuria in participants with eGFR ≥ 60 ml/min/1.73m2. The highest ALP quartile had a 1.631-fold risk elevation for albuminuria with adjustment of age and sex. (95% CI, 1.158-2.297, P = 0.005. After adjustment, the highest ALP quartile had a 1.624-fold risk elevation, for renal hyperfiltration (95% CI, 1.204-2.192, P = 0.002. In addition, hyperfiltration was significantly associated with hemoglobin, triglyceride, white blood cell count, DM, smoking, and alcohol consumption (P<0.05. The relationship between serum ALP and metabolic disorders is stronger in participants with an upper-normal range of eGFR. Higher ALP levels are significantly associated with renal hyperfiltration in Korean general population.

  6. High Serum Alkaline Phosphatase, Hypercalcaemia, Race, and Mortality in South African Maintenance Haemodialysis Patients

    Duarte, Raquel; Naicker, Saraladevi


    Objective. To determine the association between serum total alkaline phosphatase (TAP) and mortality in African maintenance haemodialysis patients (MHD). Patients and Methods. The study enrolled a total of 213 patients on MHD from two dialysis centers in Johannesburg between January 2009 and March 2016. Patients were categorized into a low TAP group (≤112 U/L) versus a high TAP group (>112 U/L) based on a median TAP of 112 U/L. Results. During the follow-up period of 7 years, there were 55 (25.8%) deaths. After adjusting for cofounders such as age, other markers of bone disorder, and comorbidity (diabetes mellitus), patients in the high TAP group had significantly higher risk of death compared to patients in the low TAP group (hazard ratio, 2.50; 95% CI 1.24–5.01, P = 0.01). Similarly, serum calcium >2.75 mmol/L was associated with increased risk of death compared to patients within levels of 2.10–2.37 mmol/L (HR 6.34, 95% CI 1.40–28.76; P = 0.02). The HR for death in white patients compared to black patients was 6.88; 95% CI 1.82–25.88; P = 0.004. Conclusion. High levels of serum alkaline phosphatase, hypercalcaemia, and white race are associated with increased risk of death in MHD patients. PMID:28168054

  7. Influence of bone and soft-tissue operations on serum concentrations of growth hormone, somatomedin C and alkaline phosphatase.

    Casser, H R; Zilkens, K W; Forst, R; Brüggemann, A


    After animal experiments suggested there was an interaction between growth hormone and bone healing, our aim in this paper was to ascertain whether there were any changes or possible interaction between the serum level of growth hormone, somatomedin C and alkaline phosphatase while a fractured bone was healing. To this end, the serum concentrations of growth hormone, somatomedin C, alkaline phosphatase and calcium were ascertained both pre- and post-operatively in two groups of patients--one with bone operations, the other with soft-tissue operations--and the results were compared. Comparing the groups, we found that after bone operations there was no increase in the serum level of growth hormone, nor of somatomedin C. An increase would have implied that these two hormones are directly involved in bone regeneration. There was no change in the serum level of alkaline phosphatase or calcium after either bone or soft-tissue operations.

  8. Relationship between serum heat—stable alkaline phosphatase level and p[regnancy

    CaoGuo-Xian; DingMei-Juan; 等


    Serum heat-stable alkaline phosphatase(HSAP) level in 649 cases of normal pregnancy and 164 cases of high-risk pregnancy is measured by raioimmunoassay (RIA).The results indicate that the HSAP level in normal prenancy increased proportionally with gestation weeks(r=0.9843).In 33 cases of pregnancy induced hypertension and 21 cases of intrauterine fetal growth retardation,the HSAP level is significantly low.In 7 cases of neonatal asphyxia and 26 cases of ftal distress,the HSAP level in the mother's serum is also low.In 53 cases of intrahepatic cholestasis of pregnancy,the HSAP level in similar to those of normal pregnancy,This study illustrates that HSAP RIA can play an important role in the evaluation of placental function and fetal prognosis for cases of high-risk pregancy.

  9. Chronic Cadmium Exposure Lead to Inhibition of Serum and Hepatic Alkaline Phosphatase Activity in Wistar Rats.

    Treviño, Samuel; Andrade-García, Alejandra; Herrera Camacho, Irma; León-Chavez, Bertha Alicia; Aguilar-Alonso, Patricia; Flores, Gonzalo; Brambila, Eduardo


    Alkaline phosphatase (ALP) activity in the serum and liver from rats administered with cadmium (Cd) in drinking water was studied. After metal administration, Cd showed a time-dependent accumulation in the liver, meanwhile metallothionein had a maximum increase at 1 month, remaining in this level until the end of the study. On the other hand, serum and liver ALP activity was decreased after 3 months exposure. To determine if Cd produced an inhibition on enzyme, apo-ALP prepared from both nonexposed and exposed rats was reactivated with Zn, showing 60% more activity as compared with the enzyme isolated from nonexposed rats. In vitro assays showed that Cd-ALP was partially reactivated with Zn; however, in the presence of cadmium, Zn-ALP was completely inhibited. Kinetic studies indicate a noncompetitive inhibition by Cd; these results suggest that Cd can substitute Zn, and/or Cd can interact with nucleophilic ligands essential for the enzymatic activity.

  10. Serum alkaline phosphatase isoenzymes in laboratory beagle dogs detected by polyacrylamide-gel disk electrophoresis.

    Hatayama, Kazuhisa; Nishihara, Yoshito; Kimura, Sayaka; Goto, Ken; Nakamura, Daichi; Wakita, Atsushi; Urasoko, Yoshinaka


    Serum alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. Itoh et al. (2002) reported that a commercially available polyacrylamide-gel (PAG) disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co., Ltd., Tokyo, Japan) for identifying serum ALP isoenzymes was useful for veterinary clinicopathological diagnosis in mongrel dogs. In the present study, based on the report of Itoh et al. (2002), we tried to expand the application range of this kit to laboratory beagle dogs which are commonly used in toxicity studies. In order to identify the origin of each ALP isoenzyme, tissue ALP extracts from the liver, bone and small intestine and serum samples were treated with neuraminidase, anti-small intestinal ALP antibody, ALP inhibitor levamisole and/or wheat germ agglutinin (WGA). The main serum ALP isoenzymes in 5-month-old intact beagle dogs were bone-derived (bone and atypical ALP: corresponding to human variant bone ALP) and they tended to decrease with age. However, liver-derived ALP isoenzyme greatly increased in the serum of cholestasis model dogs. The cholestasis model dogs also had a large molecular ALP detected in the resolving gel. This ALP could be originated from intestinal ALP or corticosteroid-induced ALP (CALP), because the activity remained even after levamisole inhibition. CALP was observed in intact laboratory beagle dogs with individual differences. These results suggest that the present method is a useful tool for detecting serum ALP isoenzymes in laboratory beagle dogs and concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity.

  11. Preoperative serum alkaline phosphatase: a predictive factor for early hypocalcaemia following parathyroidectomy of primary hyperparathyroidism

    Sun Longhao; He Xianghui; Liu Tong


    Background Postoperative hypocalcemia is one of the most common complications following parathyroidectomy for primary hyperparathyroidism (PHPT).The aim of this study was to analyze the predictive value of biochemical parameters as indicators for episodes of hypocalcemia in patients undergoing parathyroidectomy for PHPT.Methods The patients with PHPT who underwent parathyroidectomy between February 2004 and February 2014 were studied retrospectively at a single medical center.The patients were divided into biochemical,clinical,and no postoperative hypocalcemia groups,based on different clinical manifestations.Potential risk factors for postoperative hypocalcemia were identified and investigated by univariate and multivariate Logistic regression analysis.Results Of the 139 cases,25 patients (18.0%) were diagnosed with postoperative hypocalcemia according to the traditional criterion.Univariate analysis revealed only alkaline phosphatase (ALP) and the small area under the curve (AUC) of receiver operating characteristics (ROC) curve for ALP demonstrates low accuracy in predicting the occurrence of postoperative hypocalcemia.Based on new criteria,22 patients were added to the postoperative hypocalcemia group and similar biochemical parameters were compared.The serum ALP was a significant independent risk factor for postoperative hypocalcemia (P=0.000) and its AUC of ROC curve was 0.783.The optimal cutoff point was 269 U/L and the sensitivity and specificity for prediction were 89.2% and 64.3%,respectively.Conclusions The risk of postoperative hypocalcemia after parathyroidectomy should be emphasized for patients with typical symptoms of hypocalcemia despite their serum calcium level is in normal or a little higher range.Serum ALP is a predictive factor for the occurrence of postoperative hypocalcemia.

  12. Prognostic Significance of Serum Alkaline Phosphatase Level in Osteosarcoma: A Meta-Analysis of Published Data

    Hai-Yong Ren


    Full Text Available Background. Serum alkaline phosphatase (SALP is commonly elevated in osteosarcoma patients. A number of studies have investigated the prognostic role of SALP level in patients with osteosarcoma but yielded inconsistent results. Method. Systematic computerized searches were performed in PubMed, Embase, and Web of Science databases for relevant original articles. The pooled hazard ratios (HRs and relative risks (RRs with corresponding confidence intervals (CIs were calculated to assess the prognostic value of SALP level. Results. Finally, 21 studies comprising 3228 patients were included. Overall, the pooled HRs of SALP suggested that elevated level had an unfavorable impact on osteosarcoma patients’ overall survival (OS (HR = 1.82; 95% CI: 1.61–2.06; p<0.001 and event-free survival (EFS (HR = 1.97; 95% CI: 1.61–2.42; p<0.001. Combined RRs of SALP indicated that elevated level was associated with presence of metastasis at diagnosis (RR = 5.55; 95% CI: 1.61–9.49; p=0.006. No significantly different results were obtained after stratified by variables of age range, cancer stage, sample size, and geographic region. Conclusion. This meta-analysis demonstrated that high SALP level is significantly associated with poor OS or EFS rate and presence of metastasis at diagnosis. SALP level is a convenient and effective biomarker of prognosis for osteosarcoma.

  13. Evaluation of serum sialic acid, heat stable alkaline phosphatase and fucose as markers of breast carcinoma.

    Patel, P S; Baxi, B R; Adhvaryu, S G; Balar, D B


    Serum levels of total sialic acid (TSA), lipid bound sialic acid (LSA), heat stable alkaline phosphatase (HSAP) and fucose were measured in 39 patients with breast carcinoma, 14 patients with benign breast diseases and 35 healthy female individuals. Elevated levels of the four biomarkers in breast carcinoma were significant when compared with controls (p less than 0.001). Fucose levels were most sensitive (71.8%), while TSA levels were most specific (64.3%) for breast carcinoma. Sensitivity and specificity were 100% when combinations of LSA with fucose and TSA with HSAP were studied respectively. LSA was significantly elevated in infiltrating duct carcinoma patients compared with lobular carcinoma (p less than 0.001). TSA, HSAP and fucose also had lower mean values in lobular carcinoma as compared to infiltrating duct carcinoma. Increase in the levels of LSA and HSAP after surgical removal of the tumor in breast carcinoma occurred prior to the clinical evidence of the recurrence. The results indicate that the combination of the markers studied might be useful in breast cancer diagnosis and treatment monitoring.

  14. A study of the association between serum bone-specific alkaline phosphatase and serum phosphorus concentration or dietary phosphorus intake.

    Haraikawa, Mayu; Tanabe, Rieko; Sogabe, Natsuko; Sugimoto, Aoi; Kawamura, Yuka; Michigami, Toshimi; Hosoi, Takayuki; Goseki-Sone, Masae


    Alkaline phosphatase (ALP) hydrolyzes a variety of monophosphate esters into phosphoric acid and alcohol at a high optimum pH (pH 8-10). Human ALPs are classified into four types: tissue-non specific (TNSALP, liver/bone/kidney), intestinal, placental, and germ cell types. Based on studies of hypophosphatasia (HPP), which is a systemic bone disease caused by the presence of either one or two pathologic mutations in ALPL that encodes TNSALP, TNSALP was suggested to be indispensable for skeletal mineralization. In this study, we explored the possibility that dietary nutrients contribute to regulate serum bone-specific ALP (BAP) activity. Serum biochemical parameters, such as serum ALP, BAP, osteocalcin, and fibroblast growth factor 23 (FGF23), were measured in healthy young subjects (n=193). Dietary nutrient intakes were measured based on 3-d food records before the day of blood examinations. The presence of a carrier of the deletion of T at nucleotide 1559 (c.1559delT), which has been reported to be the most frequent in Japanese HPP, was not detected in any subject. By the analysis of BAP activity and other biochemical parameters or dietary nutrient intakes, we obtained significant correlations between BAP activity and serum phosphorus (r=-0.165, p=0.022), calcium intake (mg/1,000 kcal/d) (r=-0.186, p=0.010), or phosphorus intake (mg/1,000 kcal/d) (r=-0.226, p=0.002). Further study on the regulation of BAP activity and calcium and/or phosphorus homeostasis will provide useful data for improving skeletal health.

  15. A reference method for measurement of alkaline phosphatase activity in human serum.

    Tietz, N W; Burtis, C A; Duncan, P; Ervin, K; Petitclerc, C J; Rinker, A D; Shuey, D; Zygowicz, E R


    We present an official AACC reference method for the measurement of alkaline phosphatase, the culmination of optimization experiments conducted by a group of independent laboratories. The details of this method and evaluation of factors affecting the measurement are described. A metal ion buffer has been incorporated that maintains optimal and constant concentrations of zinc(II) and magnesium(II) ions. Final reaction conditions are: pH (30 degrees C), 10.40 +/- 0.05; 2-amino-2-methyl-1-propanol buffer, 0.35 mol/L; 4-nitrophenyl phosphate, 16.0 mmol/L; magnesium acetate, 2.0 mmol/L; zinc sulfate, 1.0 mmol/L; and N-(2-hydroxyethyl)ethylenediaminetriacetic acid, 2.0 mmol/L.

  16. Association of alkaline phosphatase phenotypes with arthritides

    Padmini A


    Full Text Available Arthritides, a symmetrical polyarticular disease of the bone are a heterogenous group of disorders in which hereditary and environmental factors in combination with an altered immune response appear to play a causative and pathogenic role in its occurrence. Alkaline phosphatase (ALP is an enzyme found in all tissues, with particularly high concentrations of ALP observed in the liver, bile ducts, placenta, and bone.Alkaline phosphatase is an orthophosphoric monoester phosphohydrolase catalyzing the hydrolysis of organic esters at alkaline pH, indicating that alkaline phosphatase is involved in fundamental biological processes.1 The present study envisages on identifying the specific electromorphic association of alkaline phosphatase with arthritides. Phenotyping of serum samples was carried out by PAGE (Polyacrylamide gel electrophoresis following Davies (19642 protocol on 41 juvenile arthritis, 150 rheumatoid arthritis and 100 osteo arthritis apart from, 25 normal children and 100 adult healthy subjects. Phenotyping of alkaline phosphatase revealed an increase in preponderance of p+ and p++ phenotypes in juvenile, rheumatoid and osteo arthritic patients. However a significant association of these phenotypes was observed only with rheumatoid arthritis condition (c2:17.46. Similarly, a significant increase of p+ phenotypes in female rheumatoid arthritis patients was observed (c2:14.973, suggesting that the decrease in p° (tissue non specific synthesis/secretion of alkaline phosphatase could be associated with decreased mineralization and ossification process in arthritis condition.

  17. Tissue-nonspecific alkaline phosphatase is required for the calcification of collagen in serum: a possible mechanism for biomineralization.

    Price, Paul A; Toroian, Damon; Chan, Wai Si


    Previous studies have shown that the type I collagen of tendon and demineralized bone both calcify rapidly in serum. The speed, collagen matrix-type specificity, and extent of the re-calcification of demineralized bone in serum suggest that the serum calcification activity identified in these studies may participate in normal biomineralization. Because of its presence in serum and its long history of association with the normal mineralization of the collagen matrix of bone, tissue-nonspecific alkaline phosphatase (TNAP) is an obvious candidate for a protein that could be a component of serum calcification activity, and experiments were therefore carried out to test this possibility. These experiments show that the inactivation of TNAP in serum prevents collagen calcification, and that the addition of physiological levels of purified TNAP restores the ability of TNAP-deficient serum to calcify collagen. Additional experiments show that the role of TNAP in collagen calcification is to activate a serum nucleator of apatite crystal formation. Based on these and earlier studies, the mechanism of collagen calcification in serum requires at least four elements as follows. 1) A matrix (collagen fibrils) that is accessible to small apatite crystals but not large molecules ( Toroian, D., Lim, J. E., and Price, P. A. (2007) J. Biol. Chem. 282, 22437-22447 ). 2) A large serum nucleator that generates small crystals, some of which diffuse into the fibrils. 3) A source of TNAP to activate the serum nucleator. 4) A large protein (fetuin) that selectively inhibits growth of crystals remaining in solution, thereby ensuring that only crystals within fibrils grow ( Toroian, D. T., and Price, P. A. (2008) Calcif. Tissue Int. 82, 116-126 ).

  18. Serum Proteins and Alkaline Phosphatase Levels in Patients with Tuberous Sclerosis

    Fischer, M. H.; And Others


    Six 4- to 37-year-old patients with tuberosis sclerosis (a chronic condition characterized by siezures, intercranial calcification, a reddish-yellow sebaceous glandular mass on the face, and frequent crises in early years), did not exhibit an elevation of the (alpha + beta) globulin fraction in their serum. (Author/MC)

  19. Pre-treatment serum lactate dehydrogenase and alkaline phosphatase as predictors of metastases in extremity osteosarcoma

    Leonard C. Marais


    Conclusion: In cases of osteosarcoma with LDH >850 IU/L and/or ALP >280 IU/L it may be prudent to consider more sensitive staging investigations for detection of skeletal metastases. Further research is required to determine the value and the most sensitive cut-off points of serum ALP and LDH in the prediction of skeletal metastases.

  20. [Alkaline phosphatase in Amoeba proteus].

    Sopina, V A


    In free-living Amoeba proteus (strain B), 3 phosphatase were found after disc-electrophoresis of 10 microg of protein in PAGE and using 1-naphthyl phosphate as a substrate a pH 9.0. These phosphatases differed in their electrophoretic mobilities - "slow" (1-3 bands), "middle" (one band) and "fast" (one band). In addition to 1-naphthyl phosphate, "slow" phosphatases were able to hydrolyse 2-naphthyl phosphate and p-nitrophenyl phosphate. They were slightly activated by Mg2+, completely inhibited by 3 chelators (EDTA, EGTA and 1,10-phenanthroline), L-cysteine, sodium dodecyl sulfate and Fe2+, Zn2+ and Mn2+ (50 mM), considerably inactivated by orthovanadate, molybdate, phosphatase inhibitor cocktail 1, p-nitrophenyl phosphate, Na2HPO4, DL-dithiothreitol and urea and partly inhibited by H2O2, DL-phenylalanine, 2-mercaptoethanol, phosphatase inhibitor cocktail 2 and Ca2+. Imidazole, L-(+)-tartrate, okadaic acid, NaF and sulfhydryl reagents -p-(hydroxy-mercuri)benzoate and N-ethylmaleimide - had no influence on the activity of "slow" phosphatases. "Middle" and "fast" phosphatases, in contrast to "slow" ones, were not inactivated by 3 chelators. The "middle" phosphatase differed from the "fast" one by smaller resistance to urea, Ca2+, Mn2+, phosphates and H2O2 and greater resistance to dithiothreitol and L-(+)-tartrate. In addition, the "fast" phosphatase was inhibited by L-cysteine but the "middle" one was activated by it. Of 5 tested ions (Mg2+, Cu2+, Mn2+, Ca2+ and Zn2+), only Zn2+ reactivated "slow" phosphatases after their inactivation by EDTA treatment. The reactivation of apoenzyme was only partial (about 35 %). Thus, among phosphatases found in amoebae at pH 9.0, only "slow" ones are Zn-metalloenzymes and may be considered as alkaline phosphatases (EC It still remains uncertain, to which particular phosphatase class "middle" and "fast" phosphatases (pH 9.0) may belong.

  1. Profile of serum alkaline phosphatase after inoculation of mononuclear cells and bone morphogenetic protein in the repair of osteochondral defects in rabbits

    Luiz Augusto de Souza


    Full Text Available In this study, serum alkaline phosphatase activity was measured in response to the repair of osteochondral defects in twenty-four New Zealand rabbits. The animals were divided into three groups: a control (GC, those treated with bone marrow mononuclear cells (GCM and those that received mononuclear cells with autologous bone morphogenetic protein (BMP + GCM. After exposing the trochlear groove of the left stifle joint, a wedge-shaped segment was removed. Later, the defect was filled with an osteochondral autograft preserved in 98% glycerin. For the GC group, only the bone graft was performed. For the GCM, in addition to the graft, 2x106 seed mononuclear cells were implanted. For the GCM + BMP, the same number of cells, associated with 1μg of bone morphogenetic protein, were intraarticularly administered. The osteoblastic response was measured by analyzing the serum alkaline phosphatase on day 0 (preoperative 3, 15, 30, and 45 after surgery, and by radiographic examinations. Analysis of variance in randomized blocks, factorial and Tukey’s test (p = 0.05 were made. The overall mean GCM was superior to the other groups and the highest rates were among the 15th and 45th days postoperatively. The discrepancy in values between individuals of the same group casts doubts on the veracity of the test.

  2. Bone alkaline phosphatase and mortality in dialysis patients

    C. Drechsler; M. Verduijn; S. Pilz; R.T. Krediet; F.W. Dekker; C. Wanner; M. Ketteler; E.W. Boeschoten; V. Brandenburg


    Serum alkaline phosphatase (AP) is associated with vascular calcification and mortality in hemodialysis patients, but AP derives from various tissues of origin. The aim of this study was to assess the effect of bone-specific AP (BAP) on morbidity and mortality in dialysis patients. From a prospectiv

  3. Direct Promotion of Collagen Calcification by Alkaline Phosphatase


    Alkaline phosphatase promotes hydrolysis of phosphate containing substrates, causes a rise in inorganic phosphate and, therefore, enhances calcification of biological tissues. In this work, the calcification of collagen in a model serum was used as a model of collagenous tissue biomaterials to study the possible calcification promotion mechanism of alkaline phosphatase. In the enzyme concentration range of 0.10.5mg/mL, the enzyme shows a direct calcification promoting effect which is independent of the hydrolysis of its phosphate containing substrates but proportional to the enzyme concentration. Potassium pyrophosphate somewhat inhibits the calcification promotion.

  4. Prophylactic treatment with alkaline phosphatase in cardiac surgery induces endogenous alkaline phosphatase release

    Kats, Suzanne; Brands, Ruud; Hamad, Mohamed A. Soliman; Seinen, Willem; Schamhorst, Volkher; Wulkan, Raymond W.; Schoenberger, Jacques P.; van Oeveren, Wim


    Introduction: Laboratory and clinical data have implicated endotoxin as an important factor in the inflammatory response to cardiopulmonary bypass. We assessed the effects of the administration of bovine intestinal alkaline phosphatase (bIAP), an endotoxin detoxifier, on alkaline phosphatase levels

  5. 骨碱性磷酸酶与碱性磷酸酶在佝偻病诊断中的比较%Bone alkaline phosphatase versus serum alkaline phosphatase in the diagnosis of rickets

    林岚; 陈力; 张春玲; 杨延萍; 肖玉联


    Objective To explore the significance of quantitative and qualitative measurements of bone alkaline phosphatase ( BAP-E and BAP-CIT ) and detection of serum AP in the diagnosis of rickets. Methods Serum AP level was measured, quantitative BAP was detected by ELISA, and qualitative BAP was determined by CIT in 55 children with rickets and 26 control subjects. Results AP level and BAP-E were ( 164.38 ± 23.87 ) U/L and ( 57.99 ± 7.67 ) U/L in controls, and were ( 209.50 ± 44.28 ) U/L, ( 80.38 ± 15.75 ) U/L, ( 206.06 ± 30.80 ) U/L, ( 78.05± 11.69) U/L, ( 1000.47 ± 464.65) U/L, and (283.22 ± 96.42) U/L in mild or severe rickets ,respectively. AP and BAP-E were elevated significantly in children with rickets ( P< 0.05 ), and there were significant differences between the severe groups and the mild groups ( P < 0.05 ). AP positively related with BAP-E ( r= 0.921, P< 0.01 ) and BAP-CIT ( r= 0.49, P< 0.01 ). In the control group,BAP-CIT within the normal range was only in one patient and that beyond the normal range was in the remaining patients. Conclusions AP and BAP-E are positively correlated with the severity of rickets.AP may be used in severe patients. BAP-CIT should be used carefully in the diagnosis of rickets.%目的 了解定量骨碱性磷酸酶( BAP-E)、定性骨碱性磷酸酶(BAP-CIT)、血清碱性磷酸酶(AP)三个指标在佝偻病诊断中的意义。方法 检测55例佝偻病患儿、26例对照婴幼儿血清AP、BAP-E、BAP-CIT,BAP-E采用ELISA法,BAP-CIT用全血干化学和免疫浓缩技术。结果 佝偻病患儿的AP和BAP-E较对照组(164.38 U/L±23.87 U/L,57.99 U/L±7.67 U/L)明显增高(P<0.05),中、重度组佝偻病患儿的AP和BAP-E( 1000.47 U/L±464.65U/L,283.22 U/L±96.42 U/L)较轻度组(209.50 U/L±44.28 U/L,80.38 U/L±15.75 U/L;206.06 U/L±30.80 U/L,78.05 U/L±11.69 U/L)明显增高(P<0.05),对照组BAP-CIT仅1例在试剂盒的正常范围≤200 U/L,余皆在异常范围(200-300) U/L,AP

  6. 妊娠妇女不同孕期血清碱性磷酸酶变化%Variation of serum alkaline phosphatase in pregnant in different stage



    Objective To explore the variation of serum alkaline phosphatase in Pregnancy in dif‐ferent stage .Methods Unicel Dxc800 fully automatic biochemical anaylzer was used to detect the con‐centrations of serum ALP in 522 pregnant and 157 control .Results The concentrations of serum ALP in control group ,early pregnancy ,midtrimester and late pregnancy group were (66 .58 ± 44 .09) U /L , (61 .64 ± 17 .86)U /L ,(79 .98 ± 26 .38)U /L ,and (202 .89 ± 68 .94 )U /L respectively .Statistic analysis dem‐onstrated that there was no difference between early pregnancy group and control one( P > 0 .05) .The concentrations of serum ALP in midtrimester pregnancy and late pregnancy groups were hiher than those in early pregnancy and control groups significantly ,and the concentration of serum ALP in late pregnancy group was hiher than that in midtrimester pregnancy group significantly ( P < 0 .05) .Condu‐sion The serum level of alkaline phosphatase in pregnant increased with gestational prolongation .%①目的探讨不同孕周妊娠期妇女血清碱性磷酸酶的变化。②方法采用贝克曼(Unicel Dxc800)全自动生化分析仪检测522例妊娠早、中、晚期妇女血清碱性磷酸酶值,同时选取同期157例健康育龄非妊娠妇女作为对照,方差分析不同组间血清碱性磷酸酶值差异。③结果健康育龄非妊娠妇女及妊娠期妇女孕早期、孕中期和孕晚期血清碱性磷酸酶浓度分别为(66.58±44.09)U /L 、(61.64±17.86)U /L 、(79.98±26.38)U /L 、(202.89±68.94)U /L 。统计学分析结果表明:孕早期血清碱性磷酸酶水平与对照组比较差异无统计学意义( P >0.05);孕中期、孕晚期高于孕早期及对照组,孕晚期高于孕中期,差异均具有统计学意义( P 均<0.05)。④结论妇女孕期血清碱性磷酸酶水平随孕周数增加呈逐渐上升趋势。

  7. Gallium nitrate inhibits alkaline phosphatase activity in a differentiating mesenchymal cell culture.

    Boskey, A L; Ziecheck, W; Guidon, P; Doty, S B


    The effect of gallium nitrate on alkaline phosphatase activity in a differentiating chick limb-bud mesenchymal cell culture was monitored in order to gain insight into the observation that rachitic rats treated with gallium nitrate failed to show the expected increase in serum alkaline phosphatase activity. Cultures maintained in media containing 15 microM gallium nitrate showed drastically decreased alkaline phosphatase activities in the absence of significant alterations in total protein synthesis and DNA content. However, addition of 15 microM gallium nitrate to cultures 18 h before assay for alkaline phosphatase activity had little effect. At the light microscopic and electron microscopic level, gallium-treated cultures differed morphologically from gallium-free cultures: with gallium present, there were fewer hypertrophic chondrocytes and cartilage nodules were flatter and further apart. Because of altered morphology, staining with an antibody against chick cartilage alkaline phosphatase appeared less extensive; however, all nodules stained equivalently relative to gallium-free controls. Histochemical staining for alkaline phosphatase activity was negative in gallium-treated cultures, demonstrating that the alkaline phosphatase protein present was not active. The defective alkaline phosphatase activity in cultures maintained in the presence of gallium was also evidenced when cultures were supplemented with the alkaline phosphatase substrate, beta-glycerophosphate (beta GP). The data presented suggest that gallium inhibits alkaline phosphatase activity in this culture system and that gallium causes alterations in the differentiation of mesenchymal cells into hypertrophic chondrocytes.

  8. Second generation of pseudotype-based serum neutralization assay for Nipah virus antibodies: sensitive and high-throughput analysis utilizing secreted alkaline phosphatase.

    Kaku, Yoshihiro; Noguchi, Akira; Marsh, Glenn A; Barr, Jennifer A; Okutani, Akiko; Hotta, Kozue; Bazartseren, Boldbaatar; Fukushi, Shuetsu; Broder, Christopher C; Yamada, Akio; Inoue, Satoshi; Wang, Lin-Fa


    Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and HeV-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2μl) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity.

  9. Cationized dextran nanoparticle-encapsulated CXCR4-siRNA enhanced correlation between CXCR4 expression and serum alkaline phosphatase in a mouse model of colorectal cancer

    Abedini F


    Full Text Available Fatemeh Abedini,1 Hossein Hosseinkhani,2 Maznah Ismail,1,3 Abraham J Domb,4 Abdul Rahman Omar,1,5 Pei Pei Chong,1,2 Po-Da Hong,3 Dah-Shyong Yu,6 Ira-Yudovin Farber41Laboratory of Molecular Biomedicine, Institute of Bioscience, Universiti Putra Malaysia, Selangor, 2Graduate Institute of Biomedical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan, 3Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, Selangor, Malaysia, 4Institute of Drug Research, The Center for Nanoscience and Nanotechnology, School of Pharmacy-Faculty of Medicine, The Hebrew University of Jerusalem, Jerusalem, Israel, 5Faculty of Veterinary Medicine, Universiti Putra Malaysia, Selangor, Malaysia, 6Nanomedicine Research Center, National Defense Medical Center, Taipei, TaiwanPurpose: The failure of colorectal cancer treatments is partly due to overexpression of CXCR4 by tumor cells, which plays a critical role in cell metastasis. Moreover, serum alkaline phosphatase (ALP levels are frequently elevated in patients with metastatic colorectal cancer. A polysaccharide, dextran, was chosen as the vector of siRNA. Spermine was conjugated to oxidized dextran by reductive amination process to obtain cationized dextran, so-called dextran-spermine, in order to prepare CXCR4-siRNAs/dextran-spermine nanoparticles. The fabricated nanoparticles were used in order to investigate whether downregulation of CXCR4 expression could affect serum ALP in mouse models of colorectal cancer.Methods: Colorectal cancer was established in BALB/C mice following injection of mouse colon carcinoma cells CT.26WT through the tail vein. CXCR4 siRNA for two sites of the target gene was administered following injection of naked siRNA or siRNA encapsulated into nanoparticles.Results: In vivo animal data revealed that CXCR4 silencing by dextran-spermine nanoparticles significantly downregulated CXCR4 expression compared with naked CXCR4 siRNA. Furthermore, there was

  10. [Leucocyte alkaline phosphatase in normal and pathological pregnancy (author's transl)].

    Stark, K H; Zaki, I; Sobolewski, K


    The activities of leucocyte alkaline phosphatase were determined in 511 patients with normal and pathological pregnancy. Mean values were compared and the enzyme followed up, and the conclusion was drawn that leucocyte alkaline phosphatase was no safe indicator of foetal condition. No direct relationship were found to exist between leucocyte alkaline phosphatase, total oestrogens, HSAP, HLAP, HPL, and oxytocinase.

  11. Effect Modifying Role of Serum Calcium on Mortality-Predictability of PTH and Alkaline Phosphatase in Hemodialysis Patients: An Investigation Using Data from the Taiwan Renal Registry Data System from 2005 to 2012.

    Yen-Chung Lin

    Full Text Available Predicting mortality in dialysis patients based on low intact parathyroid hormone levels is difficult, because aluminum intoxication, malnutrition, older age, race, diabetes, or peritoneal dialysis may influence these levels. We investigated the clinical implications of low parathyroid hormone levels in relation to the mortality of dialysis patients using sensitive, stratified, and adjusted models and a nationwide dialysis database. We analyzed data from 2005 to 2012 that were held on the Taiwan Renal Registry Data System, and 94,983 hemodialysis patients with valid data regarding their intact parathyroid levels were included in this study. The patient cohort was subdivided based on the intact parathyroid hormone and alkaline phosphatase levels. The mean hemodialysis duration within this cohort was 3.5 years. The mean (standard deviation age was 62 (14 years. After adjusting for age, sex, diabetes, the hemodialysis duration, serum albumin levels, hematocrit levels, calcium levels, phosphate levels, and the hemodialysis treatment adequacy score, the single-pool Kt/V, the crude and adjusted all-cause mortality rates increased when alkaline phosphatase levels were higher or intact parathyroid hormone levels were lower. In general, at any given level of serum calcium or phosphate, patients with low intact parathyroid hormone levels had higher mortality rates than those with normal or high iPTH levels. At a given alkaline phosphatase level, the hazard ratio for all-cause mortality was 1.33 (p 9.5 mg/dL, but in the group with intact parathyroid hormone levels > 300 pg/mL and serum calcium levels > 9.5 mg/dL, the hazard ratio was 0.92 (95% confidence interval 0.85-1.01. Hence, maintaining albumin-corrected high serum calcium levels at > 9.5 mg/dL may correlate with poor prognoses for patients with low intact parathyroid hormone levels.

  12. 19416例健康成人碱性磷酸酶活性分析%19416 cases of serum alkaline phosphatase activity analysis of healthy adults

    吉利; 连丽丽; 孙淑艳; 孙小慧


    目的 分析健康成人血清ALP活性,探讨ALP水平在成人性别和年龄的分布规律.方法 随机选择2008年来吉大一院健康体检各项指标正常的健康人19416例,对其血清ALP活性进行回顾性统计分析.结果 21-50岁男性ALP值显著高于同年龄组女性(P<0.01);51岁以上男性ALP值显著低于同年龄段女性(P<0.01).男性20-30岁ALP水平显著高于31岁以上各年龄组,31岁以上各年龄组间差异无统计学意义;女性20-40岁年龄段ALP水平差异无统计学意义,40岁以后,ALP水平随年龄增长而增高.结论 ALP在性别和年龄的分布规律,反映了健康成人骨代谢情况,雌激素可降低ALP活性,为研究雌激素对成骨细胞标志物ALP的影响提供科学的依据.%bjective To Analyze the activity of Alkaline phosphatase (ALP) in serum of healthy adults ,in terms of discussing the distribution rule of ALP levels in gender and age .Methods Making retrospective analysis of the ALP activity data of healthy people who has normal medical examination results from the physical examination center our hospital in 2008 .Results The ALP levels of 21-30,31-40 and 41-50 age group male were significantly higher than female of the same age group ( P<0.01 ) ;the ALP levels of 51-60 and older than 60 age groups male were significantly lower than those of the same age female ( P<0.01 ) .The variance analysis results of ALP levels between different age groups of male and female respectively showed that the ALP levds of 21-30 yearold male were obviously hgher than those of all age groups above 31 year-old and the male ALP results above 31 age groups has no different significance in statistics ;the ALP levels of 20-40 year-old female has no different significance in statistics and the other age groups has statistical significance the female ALP levels increased from 40 year-old as age grew .Conclusion Ths research obtained the distribution rule of adults serum ALP activity in gender and

  13. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Jemmerson, R.; Low, M.G.


    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  14. Kinetic aspects of human placental alkaline phosphatase enzyme membrane.

    Roig, M G; Serrano, M A; Bello, J F; Cachaza, J M; Kennedy, J F


    The crosslinking of alkaline phosphatase of human placenta with human serum albumin has been optimized. During the physico-chemical characterization of this immobilized biocatalyst, special attention was paid to attributes such as the irreversibility of the enzyme support bonding, the stability of the catalytic activity, and the effects of pH and temperature on this activity. Regarding stability, patterns of denaturation are proposed, to account for inactivation curves over time and under storage/operation conditions. These patterns, in some cases, indicate the existence of different populations of immobilized enzyme molecules, with a different degree of sensitivity to denaturation. The activity vs pH profiles are clearly modified by the immobilization process. This is because the pH of the free homogeneous solution, measurable with a pH-meter, differs from the real pH of the immediate microenvironment of the immobilized enzyme molecules due to the effects of proton accumulation in the microenvironment (in the reaction catalysed by alkaline phosphatase, protons are produced), to limitations to the free diffusion of H+ and to the possible partition effects of H+ due to polar interactions with residues or molecules of the enzyme membrane. In the experimental working conditions, the apparent optimum temperatures are centered at 40 degrees C, inactivation (thermal denaturation) occurring above this temperature. In the temperature range 10-40 degrees C, the kinetic control over the overall activity of the immobilized enzyme was observed, causing the Arrhenius profiles to be linear.

  15. Measurement of bone alkaline phosphatase and relative study with osteosarcoma

    YANG Zhiping; HUO Yanqing; SUN Guangzhi; LI Jianmin; LI Xin


    The objective of this paper is to explore the value of bone alkaline phosphatase (BALP) for diagnosing osteosarcoma,evaluating the effect of the chemotherapy,judging the prognosis and supervising the relapse and metastasis.The immunoassay was used to check the BALP of the blood serum that was from 42 primary osteosarcoma patients.Alkaline phosphatase (ALP) in blood serum was checked with auto biochemistry equipment.The biopsy tissue and the lesion resected in operation were treated with pathology and histological response was counted.The patients were followed up from five months to 49 months with an average of 24.3 months.Eighteen cases relapsed and transferred,among which,16 of them were dead,and others were survival to the end of the follow-up.BALP was more sensitive than ALP in diagnosing osteosarcoma (P = 0.015).Fifteen cases decreased to normal value in ALP after preoperative chemotherapy,and 34 cases decreased in BALP.Both ALP and BALP in all cases decreased to normal value in postoperative.There was significant difference in positive correlation between the decrease of BALP and the increase of histological response (P = 0.001,r = 0.642).In the followup,there was significant difference in BALP between the group of relapse and transfer and the group of free disease survival (P=0.000).As a check marker in blood serum,BALP,reflecting the process of ossification,has a higher sensitivity than ALP.It has applied value in the diagnosis of osteosarcoma,reflection of the effect of chemotherapy and forecast the prognosis.

  16. The catalytic properties of alkaline phosphatases under various conditions

    Atyaksheva, L. F.; Chukhrai, E. S.; Poltorak, O. M.


    A comparative study was performed to examine the catalytic properties of alkaline phosphatases from bacteria Escherichia coli and bovine and chicken intestines. The activity of enzyme dimers and tetramers was determined. The activity of the dimer was three or four times higher than that of the tetramer. The maximum activity and affinity for 4-nitrophenylphosphate was observed for the bacterial alkaline phosphatase ( K M = 1.7 × 10-5 M, V max = 1800 μmol/(min mg of protein) for dimers and V max = 420 μmol/(min mg of protein) for tetramers). The Michaelis constants were equal for two animal phosphatases in various buffer media (pH 8.5) ((3.5 ± 0.2) × 10-4 M). Five buffer systems were investigated: tris, carbonate, hepes, borate, and glycine buffers, and the lowest catalytic activity of alkaline phosphatases at equal pH was observed in the borate buffer (for enzyme from bovine intestine, V max = 80 μmol/(min mg of protein)). Cu2+ cations formed a complex with tris-(oxymethyl)-aminomethane ( tris-HCl buffer) and inhibited the intestine alkaline phosphatases by a noncompetitive mechanism.

  17. Human placental alkaline phosphatase electrophoretic alleles: Quantitative studies

    Lucarelli, Paola; Scacchi, Renato; Corbo, Rosa Maria; Benincasa, Alberto; Palmarino, Ricciotti


    Human placental alkaline phosphatase (ALP) activity has been determined in specimens obtained from 562 Italian subjects. The mean activities of the three common homozygotes (Pl 2 = 4.70 ± 0.24, Pl 1 = 4.09 ± 0.08, and Pl 3 = 2.15 ± 0.71 μmol of p-nitrophenol produced) were significantly different. The differences among the various allelic forms account for 10% of the total quantitative variation of the human placental alkaline phosphatase. PMID:7072721

  18. Induction of glomerular alkaline phosphatase after challenge with lipopolysaccharide

    Kapojos, Jola Jovita; Poelstra, Klaas; Borghuis, Theo; van den Berg, Anke; Baelde, Hans J.; Klok, P.A; Bakker, W.W


    Alkaline phosphatase (AP) can be considered as a host defence molecule since this enzyme is able to detoxify bacterial endotoxin at physiological pH. The question emerged whether this anti-endotoxin principle is inducible in the glomerulus and if so, which glomerular cells might be involved in the e

  19. Chromatographic separation of alkaline phosphatase from dental enamel

    Moe, D; Kirkeby, S; Salling, E


    Alkaline phosphatase (AP) was prepared from partly mineralized bovine enamel by extraction in phosphate buffer, centrifugation and various chromatographic techniques. Chromatofocusing showed that the enamel enzyme possessed five isoelectric points at the acid pH level ranging from pH 5.7 to pH 4....

  20. Dephosphorylation of endotoxin by alkaline phosphatase in vivo

    Poelstra, Klaas; Bakker, W.W; Klok, P.A; Kamps, J.AAM; Hardonk, M.J; Meijer, D.K F


    Natural substrates for alkaline phosphatase (AP) are at present not identified despite extensive investigations. Difficulties in imagining a possible physiological function involve its extremely high pH optimum for the usual exogenous substrates and its localization as an ecto-enzyme. As endotoxin i

  1. Endotoxin detoxification by alkaline phosphatase in cholestatic livers

    Poelstra, K; Bakker, WW; Hardonk, MJ; Meijer, DKF; Wisse, E; Knook, DL; Balabaud, C


    Increased expression of alkaline phosphatase (AP) in the liver is a hallmark of cholestasis but the pathophysiological role of this is not clear. We argue that deprotonation of carboxyl groups at the active site of the enzyme may be a prerequisite for optimal AP activity. Such a creation of negative

  2. Observations on the alkaline phosphatase isoenzyme distribution in maternal and amniotic fluid compartments in Nigerian parturients.

    Okpere, E; Okorodudu, A; Gbinigie, O


    Estimation of the alkaline phosphates isoenzymes in paired maternal serum and amniotic fluids in term uncomplicated pregnancies and in patients with pre-eclampsia, showed poor correlation coefficients between the levels of both heat stable and heat labile isoenzymes. There was a statistically significant fall in AF (P less than .05) HSAP in pre-eclampsia and a highly significant rise of HLAP in meconial liquor. It is concluded that the poor correlation between the levels of HSAP in maternal serum and amniotic fluid (despite their common source of origin), the normal levels of HLAP in maternal serum in the presence of significantly high levels of HSAP in maternal serum in the presence of significantly diminished levels in amniotic fluid point to a state of relatively diminished permeability of the chorioamniotic membranes to the alkaline phosphatase isoenzymes in Nigerians.

  3. Activation of Calf Intestinal Alkaline Phosphatase by Trifluoroethanol

    曹志方; 徐真; 朴龙斗; 周海梦


    Alkaline phosphatase is a stable enzyme which is strongly resistant to urea, guanidine hydrochloride, acid pH, and heat. But there have been few studies on the effect of organic cosolvents on the activity and structure of alkaline phosphatase. The activity of calf intestinal alkaline phosphatase (CIAP) is markedly increased when incubated in solutions with elevated trifluoroethanol (TFE) concentrations. The activation is a time dependent course. There is a very fast phase in the activation kinetics in the mixing dead time (30 s) using convential methods. Further activation after the very fast phase follows biphasic kinetics. The structural basis of the activation has been monitored by intrinsic fluorescence and far ultraviolet circular dichroism. TFE (0 - 60%) did not lead to any significant change in the intrinsic fluorescence emission maximum, indicating no significant change in the tertiary structure of CIAP. But TFE did significantly change the secondary structure of CIAP, especially increasing α-helix content. We conclude that the activation of ClAP is due to its secondary structural change. The time for the secondary structure change induced by TFE preceds that of the activity increase. These results suggest that a rapid conformational change of ClAP induced by TFE results in the enhancement of ClAP activity, followed by further increase of this activity because of the further slightly slower rearrangements of the activated conformation. It is concluded that the higher catalytic activity of ClAP can be attained with various secondary structures.

  4. Combination of alkaline phosphatase anti-alkaline phosphatase (APAAP)- and avidin-biotin-alkaline phosphatase complex (ABAP)-techniques for amplification of immunocytochemical staining of human testicular tissue.

    Davidoff, M S; Schulze, W; Holstein, A F


    An amplification procedure was developed for the visualization of antigens in human testis using monoclonal antibodies against desmin and vimentin. The technique combines the high sensitive and specific APAAP- and ABAP-methods. Depending on the quality of the antibodies used and the processing of the material prior to the immunocytochemical staining the amplification technique may be applied either as a single APAAP and ABAP- or as a double APAAP and ABAP-combination. Especially after the double amplification reaction a distinct increase of the staining intensity of the vimentin- (in Sertoli cells, myofibroblasts of the lamina propria, and fibroblasts of the interstitium) and desmin- (in myofibroblasts of the lamina propria and smooth muscle cells of the blood vessels) like immunoreactivity was observed. If different diazonium salts were used for the visualization of the alkaline phosphatase activity (e.g. Fast Red TR Salt, Fast Blue BB Salt) desmin- and vimentin-like immunoreactivity can be demonstrated in the same tissue section in a double sequential staining approach. For double staining, the alkaline phosphatase technique may be combined successfully with a technique or a combination that uses peroxidase as a marker.

  5. phoD Alkaline Phosphatase Gene Diversity in Soil.

    Ragot, Sabine A; Kertesz, Michael A; Bünemann, Else K


    Phosphatase enzymes are responsible for much of the recycling of organic phosphorus in soils. The PhoD alkaline phosphatase takes part in this process by hydrolyzing a range of organic phosphoesters. We analyzed the taxonomic and environmental distribution of phoD genes using whole-genome and metagenome databases. phoD alkaline phosphatase was found to be spread across 20 bacterial phyla and was ubiquitous in the environment, with the greatest abundance in soil. To study the great diversity of phoD, we developed a new set of primers which targets phoD genes in soil. The primer set was validated by 454 sequencing of six soils collected from two continents with different climates and soil properties and was compared to previously published primers. Up to 685 different phoD operational taxonomic units were found in each soil, which was 7 times higher than with previously published primers. The new primers amplified sequences belonging to 13 phyla, including 71 families. The most prevalent phoD genes identified in these soils were affiliated with the orders Actinomycetales (13 to 35%), Bacillales (1 to 29%), Gloeobacterales (1 to 18%), Rhizobiales (18 to 27%), and Pseudomonadales (0 to 22%). The primers also amplified phoD genes from additional orders, including Burkholderiales, Caulobacterales, Deinococcales, Planctomycetales, and Xanthomonadales, which represented the major differences in phoD composition between samples, highlighting the singularity of each community. Additionally, the phoD bacterial community structure was strongly related to soil pH, which varied between 4.2 and 6.8. These primers reveal the diversity of phoD in soil and represent a valuable tool for the study of phoD alkaline phosphatase in environmental samples.

  6. Gamma-glutamyltransferase, aspartate aminotransferase and alkaline phosphatase as markers of alcohol consumption in out-patient alcoholics

    Gluud, C; Andersen, I; Dietrichson, O;


    Serum activity of gamma-glutamyltransferase, aspartate aminotransferase and alkaline phosphatase were determined in 316 patients attending an out-patients clinic for treatment of alcoholism. The activity of gamma-glutamyltransferase was raised in 34% and that of aspartate aminotransferase and alk...

  7. Growth hormone (GH) treatment increases serum insulin-like growth factor binding protein-3, bone isoenzyme alkaline phosphatase and forearm bone mineral content in young adults with GH deficiency of childhood onset

    Juul, A; Pedersen, S A; Sørensen, S;


    the effect of GH treatment on a marker of bone formation (bone alkaline phosphatase), hepatic excretory function and distal forearm bone mineral content in GH-deficient adults. Growth hormone was administered subcutaneously in 21 adults (13 males and 8 females) with GH deficiency of childhood onset for 4......Recent studies have demonstrated that growth hormone (GH)-deficient adults have a markedly decreased bone mineral content compared to healthy adults. However, there are conflicting results regarding the effects of GH treatment on bone mineral content in GH-deficient adults. Therefore, we evaluated...

  8. Growth hormone (GH) treatment increases serum insulin-like growth factor binding protein-3, bone isoenzyme alkaline phosphatase and forearm bone mineral content in young adults with GH deficiency of childhood onset

    Juul, A; Pedersen, S A; Sørensen, S;


    Recent studies have demonstrated that growth hormone (GH)-deficient adults have a markedly decreased bone mineral content compared to healthy adults. However, there are conflicting results regarding the effects of GH treatment on bone mineral content in GH-deficient adults. Therefore, we evaluated...... the effect of GH treatment on a marker of bone formation (bone alkaline phosphatase), hepatic excretory function and distal forearm bone mineral content in GH-deficient adults. Growth hormone was administered subcutaneously in 21 adults (13 males and 8 females) with GH deficiency of childhood onset for 4...




    Full Text Available The investigation was performed to evaluate the dog semen freezability and itsquality after thawing allowing its use for artificial insemination (AI. On the basis ofsperm motility, concentration and alkaline phosphatase (AP activity in semenplasma it was possible to establish that AP activity corresponds with the basic factorof semen examination. Significant statistical differences occurred between thequality of ejaculates which were qualified or disqualified to deep freezing and AI.These results show that AP activity in raw dog semen plasma can be used as amarker for the dog semen qualification for deep freezing and AI with 95%probability of the prognosis of the results.

  10. Plasma calcium, magnesium, phosphorus, and alkaline phosphatase levels in normal British schoolchildren.

    Round, J M


    In a cross-sectional survey 624 schoolchildren were screened for plasma calcium, inorganic phosphate, and alkaline phosphatase levels. Plasma magnesium and alkaline phosphatase isoenzymes were also estimated in some cases.No significant difference was found between adult and childhood values for calcium and magnesium. Levels of alkaline phosphatase and inorganic phosphorus varied with both age and sex. The magnitude of these variations in normal ranges is of clear importance in assessing data from individual paediatric or adolescent patients.

  11. Effect of carbon source on alkaline phosphatase production and excretion in Aspergillus caespitosus.

    Guimarães, Luis Henrique Souza; Jorge, João Atilio; Terenzi, Héctor Francisco; Jamur, Maria Célia; Oliver, Constance; De Lourdes Teixeira De Moraes Polizeli, Maria


    The effect of several carbon sources on the production of alkaline phosphatase by the thermotolerant Aspergillus caespitosus was analysed. The fungus released high levels of alkaline phosphatases into the medium after being cultured for long periods with xylan or industrial residues such as wheat raw and sugar cane bagasse in the culture media. In contrast, the alkaline phosphatase activities were found only intracellulary when the fungus was cultured in glucose-supplemented media. The pH of the medium likely affects the process of enzyme secretion according to the carbon source used. Addition of xylan or industrial residues in the culture medium stimulated the secretion of phosphatases. In contrast, media supplemented with glucose or disaccharides promoted retention of these enzymes into the cells. The subcellular location activities of alkaline phosphatases were studied using histochemical and immunochemical methods and showed that alkaline phosphatases were present in the mycelial walls and septa.

  12. Establishing Quantitative Standards for Residual Alkaline Phosphatase in Pasteurized Milk

    Chon, Jung-Whan; Kim, Hyunsook; Kim, Kwang-Yup


    The alkaline phosphatase (ALP) assay is a rapid and convenient method for verifying milk pasteurization. Since colorimetric ALP assays rely on subjective visual assessments, their results are especially unreliable near the detection limits. In this study, we attempted to establish quantitative criteria for residual ALP in milk by using a more objective method based on spectrophotometric measurements. Raw milk was heat-treated for 0, 10, 20, 30, and 40 min and then subjected to ALP assays. The quantitative criteria for residual ALP in the milk was determined as 2 μg phenol/mL of milk, which is just above the ALP value of milk samples heat-treated for 30 min. These newly proposed methodology and criteria could facilitate the microbiological quality control of milk. PMID:27194927

  13. Intestinal alkaline phosphatase prevents metabolic syndrome in mice.

    Kaliannan, Kanakaraju; Hamarneh, Sulaiman R; Economopoulos, Konstantinos P; Nasrin Alam, Sayeda; Moaven, Omeed; Patel, Palak; Malo, Nondita S; Ray, Madhury; Abtahi, Seyed M; Muhammad, Nur; Raychowdhury, Atri; Teshager, Abeba; Mohamed, Mussa M Rafat; Moss, Angela K; Ahmed, Rizwan; Hakimian, Shahrad; Narisawa, Sonoko; Millán, José Luis; Hohmann, Elizabeth; Warren, H Shaw; Bhan, Atul K; Malo, Madhu S; Hodin, Richard A


    Metabolic syndrome comprises a cluster of related disorders that includes obesity, glucose intolerance, insulin resistance, dyslipidemia, and fatty liver. Recently, gut-derived chronic endotoxemia has been identified as a primary mediator for triggering the low-grade inflammation responsible for the development of metabolic syndrome. In the present study we examined the role of the small intestinal brush-border enzyme, intestinal alkaline phosphatase (IAP), in preventing a high-fat-diet-induced metabolic syndrome in mice. We found that both endogenous and orally supplemented IAP inhibits absorption of endotoxin (lipopolysaccharides) that occurs with dietary fat, and oral IAP supplementation prevents as well as reverses metabolic syndrome. Furthermore, IAP supplementation improves the lipid profile in mice fed a standard, low-fat chow diet. These results point to a potentially unique therapy against metabolic syndrome in at-risk humans.

  14. Alkaline phosphatase protects against renal inflammation through dephosphorylation of lipopolysaccharide and adenosine triphosphate

    Peters, E; Geraci, S; Heemskerk, S; Wilmer, M J; Bilos, A; Kraenzlin, B; Gretz, N; Pickkers, P; Masereeuw, R


    BACKGROUND AND PURPOSE: Recently, two phase-II trials demonstrated improved renal function in critically ill patients with sepsis-associated acute kidney injury treated with the enzyme alkaline phosphatase. Here, we elucidated the dual active effect on renal protection by alkaline phosphatase presum

  15. Activity of alkaline phosphatase adsorbed and grafted on "polydopamine" films.

    Ball, Vincent


    The oxidation of dopamine in slightly basic solutions and in the presence of oxygen as an oxidant allows for the deposition of dopamine-eumelanin ("polydopamine") films on almost all kinds of materials allowing for an easy secondary functionalization. Molecules carrying nucleophilic groups like thiols and amines can be easily grafted on those films. Herein we show that alkaline phosphatase (ALP), as a model enzyme, adsorbs to "polydopamine" films and part of the adsorbed enzyme is rapidly desorbed in contact with Tris buffer. However a significant part of the enzyme remains irreversibly adsorbed and keeps some enzymatic activity for at least 2 weeks whereas ALP adsorbed on quartz slides is rapidly and quantitatively deactivated. In addition we estimated the Michaelis constant Km of the enzyme irreversibly bound to the "polydopamine" film. The Michaelis constant, and hence the affinity constant between paranitrophenol phosphate and ALP are almost identical between the enzyme bound on the film and the free enzyme in solution. Complementarily, it was found that "polydopamine" films display some phosphatase like catalytic activity.

  16. Alkaline phosphatase levels in patients with coronary heart disease saliva and its relation with periodontal status

    Yunita, Dina Suci; Masulili, Sri Lelyati C.; Tadjoedin, Fatimah M.; Radi, Basuni


    Coronary heart disease (CHD) is a disease that causes narrowing of the coronary arteries. Currently, there is a hypothesis regarding periodontal infection that increases risk for heart disease. Alkaline phosphatase (ALP) as a marker of inflammation will increase in atherosclerosis and periodontal disease. The objective of this research is analyzing the relationship between the levels of alkaline phosphatase in saliva with periodontal status in patients with CHD and non CHD. Here, saliva of 104 subjects were taken, each 1 ml, and levels of Alkaline Phosphatase was analyzed using Abbott ci4100 architect. We found that no significant difference of Alkaline Phosphatase levels in saliva between CHD patients and non CHD. Therefore, it can be concluded that Alkaline Phosphatase levels in patients with CHD saliva was higher than non CHD and no association between ALP levels with periodontal status.

  17. Comparative evaluation of Schistosoma mansoni, Schistosoma intercalatum, and Schistosoma haematobium alkaline phosphatase antigenicity by the alkaline phosphatase immunoassay (APIA).

    Cesari, I M; Ballén, D E; Mendoza, L; Ferrer, A; Pointier, J-P; Kombila, M; Richard-Lenoble, D; Théron, A


    To know if alkaline phosphatase (AP) from schistosomes other than Schistosoma mansoni can be used as diagnostic marker for schistosomiasis in alkaline phosphatase immunocapture assay (APIA), we comparatively tested n-butanol extracts of adult worm membranes from a Venezuelan (JL) strain of S. mansoni (Ven/AWBE/Sm); a Cameroonian (EDEN) strain of Schistosoma intercalatum (Cam/AWBE/Si) and a Yemeni strain of Schistosoma haematobium (Yem/AWBE/Sh). APIA was evaluated with sera of patients from Venezuela, Senegal, and Gabon infected with S. mansoni, from Gabon infected with S. intercalatum or S. haematobium, from Chine infected with Schistosoma japonicum and from Cambodian patients infected with Schistosoma mekongi. Results indicate that 92.5% (37/40) of Venezuela sera, 75% (15/20) of Senegal sera, 39.5% (17/43) of S. haematobium sera, and 19.2% (5/26) S. intercalatum sera were APIA-positive with the Ven/AWBE/Sm preparation. APIA with the Cam/AWBE/Si preparation showed that 53.8% of S. intercalatum-positive sera had anti-AP antibodies, and 51.2% S. haematobium-positive sera cross-immunocapturing the S. intercalatum AP. APIA performed with Yem/AWBE/Sh showed that 55.8% S. haematobium sera were positive. Only two out of nine S. japonicum sera were APIA-positive with the Ven/AWBE/Sm and Cam/AWBE/Si, and no reaction was observed with Cambodian S. mekongi-positive sera. AP activity was shown to be present in all the schistosome species/strains studied. The use of APIA as a tool to explore the APs antigenicity and the presence of Schistosoma sp. infections through the detection of anti-Schistosoma sp. AP antibodies in a host, allowed us to demonstrate the antigenicity of APs of S. mansoni, S. intercalatum, and S. haematobium.

  18. Pediatric reference intervals of serum alkaline phosphatase for healthy Han population in Changchun%长春市汉族儿童血清碱性磷酸酶参考区间的建立

    王迪; 杨春; 周琪; 许建成


    目的:建立0~14岁长春市汉族儿童血清碱性磷酸酶(ALP)的参考区间。方法采用日立7600-210全自动生化分析仪检测4211名健康体检儿童(男2090名,女2121名)血清 ALP。弃离群值后,判断数据是否正态分布。One-Way ANOVA 比较组间差异及确定是否需性别、年龄分组。非参数方法计算参考值的2.5百分位数和97.5百分位数,R 语言计算90%置信区间。结果长春市儿童 ALP 参考区间有年龄及性别差异。婴儿出生1个月内 ALP 水平较低,此后开始增高,1个月至11岁儿童 ALP 水平相对稳定且无性别差异。12岁后儿童 ALP 水平逐渐出现性别差异,12~14岁女孩 ALP 水平逐渐下降,而12~14岁男孩 ALP 水平高于同年龄段女孩。年龄、性别合并后的参考区间包括0~30 d、1~12月、1~10岁、11岁、12~14岁(男)、12岁(女)、13岁(女)及14岁(女)。结论建立儿童年龄、性别相关的血清 ALP 参考区间对儿童预防保健及疾病状况分析具有临床应用价值。%Objective To establish pediatric reference intervals of serum alkaline phosphatase (ALP) for healthy children with an age range of 0-14 years old in Changchun. Methods A total of 4 211 healthy children (2 090 males and 2 121 females) were enrolled in Changchun. ALP was performed on Hitachi 7600-210 automatic biochemical analyzer. After outlier data exclusion, data were estimated to or not to follow Gaussian distributions. One-way ANOVA was used to compare the differences of gender and age groups. The 2.5th and 97.5th percentiles of ALP were calculated by nonparametric method and 90%confidence intervals were computed by R language. Results The study showed there were apparent age and gender variations of the reference intervals for ALP. After a temporary low level in newborns, there was an increase from 1 month. The reference intervals of ALP were relatively stable and there was no significant gender

  19. Alkaline Phosphatase Assay for Freshwater Sediments: Application to Perturbed Sediment Systems

    Sayler, Gary S.; Puziss, Marla; Silver, Martin


    The p-nitrophenyl phosphate hydrolysis-phosphatase assay was modified for use in freshwater sediment. Laboratory studies indicated that the recovery of purified alkaline phosphatase activity was 100% efficient in sterile freshwater sediments when optimized incubation and sonication conditions were used. Field studies of diverse freshwater sediments demonstrated the potential use of this assay for determining stream perturbation. Significant correlations between phosphatase and total viable cell counts, as well as adenosine triphosphate biomass, suggested that alkaline phosphatase activity has utility as an indicator of microbial population density and biomass in freshwater sediments. PMID:16345464

  20. Alkaline phosphatase in stallion semen: characterization and clinical applications.

    Turner, R M O; McDonnell, S M


    Significant amounts of alkaline phosphatase (AP) activity have been found in semen plasma from numerous species. In species in which the majority of semen plasma AP (SPAP) activity originates from the epididymis and testicle, SPAP activity can be used clinically as a marker to differentiate testicular origin azoospermia or oligospermia from ejaculatory failure. Information on SPAP activity in stallions to date has been limited. In this study, a standard clinical chemistry analyzer was used to determine AP activity in pre-ejaculatory fluid and ejaculates from groups of normal stallions. Additionally, accessory glands, epididymides, testicles and other components of the urogenital tract of normal stallions were assayed to determine which tissues contain SPAP activity. The results indicated that levels of AP activity are low in pre-ejaculatory fluid, but significantly higher in ejaculatory fluid from normal stallions. Spermatozoa were not a significant source of SPAP activity. High levels of SPAP activity were found in the testes and epididymides. These findings suggest that SPAP activity is a candidate for a sperm-independent marker for ejaculation in the stallion. Finally, AP activity was determined in ejaculatory fluid from a stallion with bilaterally blocked ampullae, both before and after relief of the blockage. While the blockage was present, AP activity in ejaculatory fluid was low. However, following relief of the blockage, AP activity in ejaculatory fluid rose dramatically, thus suggesting that AP activity will be useful as an inexpensive, simple clinical assay for differentiating ejaculatory failure or excurrent duct blockages from testicular origin azoospermia and oligospermia.

  1. Interplay between intestinal alkaline phosphatase, diet, gut microbes and immunity.

    Estaki, Mehrbod; DeCoffe, Daniella; Gibson, Deanna L


    Intestinal alkaline phosphatase (IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP's role in the intestine is to dephosphorylate toxic microbial ligands such as lipopolysaccharides, unmethylated cytosine-guanosine dinucleotides and flagellin as well as extracellular nucleotides such as uridine diphosphate. IAP's ability to detoxify these ligands is essential in protecting the host from sepsis during acute inflammation and chronic inflammatory conditions such as inflammatory bowel disease. Also important in these complications is IAP's ability to regulate the microbial ecosystem by forming a complex relationship between microbiota, diet and the intestinal mucosal surface. Evidence reveals that diet alters IAP expression and activity and this in turn can influence the gut microbiota and homeostasis. IAP's ability to maintain a healthy gastrointestinal tract has accelerated research on its potential use as a therapeutic agent against a multitude of diseases. Exogenous IAP has been shown to have beneficial effects when administered during ulcerative colitis, coronary bypass surgery and sepsis. There are currently a handful of human clinical trials underway investigating the effects of exogenous IAP during sepsis, rheumatoid arthritis and heart surgery. In light of these findings IAP has been marked as a novel agent to help treat a variety of other inflammatory and infectious diseases. The purpose of this review is to highlight the essential characteristics of IAP in protection and maintenance of intestinal homeostasis while addressing the intricate interplay between IAP, diet, microbiota and the intestinal epithelium.

  2. Dairy products and the French paradox: Could alkaline phosphatases play a role?

    Lallès, Jean-Paul


    The French paradox - high saturated fat consumption but low incidence of cardiovascular disease (CVD) and mortality - is still unresolved and continues to be a matter of debate and controversy. Recently, it was hypothesised that the high consumption of dairy products, and especially cheese by the French population might contribute to the explanation of the French paradox, in addition to the "(red) wine" hypothesis. Most notably this would involve milk bioactive peptides and biomolecules from cheese moulds. Here, we support the "dairy products" hypothesis further by proposing the "alkaline phosphatase" hypothesis. First, intestinal alkaline phosphatase (IAP), a potent endogenous anti-inflammatory enzyme, is directly stimulated by various components of milk (e.g. casein, calcium, lactose and even fat). This enzyme dephosphorylates and thus detoxifies pro-inflammatory microbial components like lipopolysaccharide, making them unable to trigger inflammatory responses and generate chronic low-grade inflammation leading to insulin resistance, glucose intolerance, type-2 diabetes, metabolic syndrome and obesity, known risk factors for CVD. Various vitamins present in high amounts in dairy products (e.g. vitamins A and D; methyl-donors: folate and vitamin B12), and also fermentation products such as butyrate and propionate found e.g. in cheese, all stimulate intestinal alkaline phosphatase. Second, moulded cheeses like Roquefort contain fungi producing an alkaline phosphatase. Third, milk itself contains a tissue nonspecific isoform of alkaline phosphatase that may function as IAP. Milk alkaline phosphatase is present in raw milk and dairy products increasingly consumed in France. It is deactivated by pasteurization but it can partially reactivate after thermal treatment. Experimental consolidation of the "alkaline phosphatase" hypothesis will require further work including: systematic alkaline phosphatase activity measurements in dairy products, live dairy ferments and

  3. Barley seed coating with free and immobilized alkaline phosphatase to improve P uptake and plant growth

    Pilar Izquierdo, María Concepción; Ortega Santamaría, Natividad; Pérez Mateos, Manuel; Busto Núñez, Mª Dolores


    Coating barley seeds with free and immobilized alkaline phosphatase was investigated as a potential means to enhance plant utilization of accumulated soil phosphorus (P). Two coating techniques were studied: film-coating and pelleting. The highest phosphatase activity retention in the coating layer, ranging from 0·48 to 0·67, was observed when seeds were film-coated with phosphatase–polyresorcinol complex (PPC). The germination of seeds film-coated or pelleted with alkaline phosph...

  4. A study on the occurrence of alkaline phosphatase in the sutura interfrontalis of Wistar rats.

    Markens, I S; Oudhof, H A


    The aim of the present study was to determine the presence of alkaline phosphatase during various stages in development and closure of the sutura interfrontalis. The histological sections reveal that this enzyme could primarily be demonstrated in the dura mater of this suture. In further developmental stages, alkaline phosphatase could be observed within the intermediate zone as well as the pericranium. These findings are brought in relation with the occurrence of synostosis which can be induced under experimental conditions.

  5. Kidney alkaline phosphatase in mercuric chloride injected chicks resistant and susceptible to leukosis

    Miller, V.L.; McIntyre, J.A.; Bearse, G.E.


    Two strains of chickens were selected for resistance and susceptibility to avian leukosis. Researchers found that the resistant chicks retained two to four times as much mercury in the liver and kidneys as did the susceptible chicks following injection of mercuric chloride or phenylmercuric acetate. Differences in alkaline phosphatase in the kidneys of the resistant and susceptible chicks, and the effect of the mercuric chloride injection on the alkaline phosphatase activity were reported in this paper. 19 references, 2 tables.

  6. Osteocalcin and bone-specific alkaline phosphatase in Asian elephants (Elephas maximus) at different ages.

    Arya, Nlin; Moonarmart, Walasinee; Cheewamongkolnimit, Nareerat; Keratikul, Nutcha; Poon-Iam, Sawinee; Routh, Andrew; Bumpenpol, Pitikarn; Angkawanish, Taweepoke


    Bone turnover markers could offer a potential alternative means for the early diagnosis of metabolic bone disease in young growing elephants although the baseline of bone turnover markers in elephant is not well established. The aim of this study was to determine any relationship between the age of captive Asian elephants (Elephas maximus) and markers of bone formation. Serum samples from 24 female Asian elephants were collected to evaluate levels of two bone formation markers, namely, osteocalcin (OC) and bone-specific alkaline phosphatase (BAP). Both intact and N-terminal midfragment OC and BAP were negatively correlated with age. The findings demonstrate that younger elephants have a higher rate of bone turnover than older elephants. Use of these and additional bone markers could lead to the establishment of validated protocols for the monitoring of bone disease in elephants.

  7. Intestinal alkaline phosphatase: selective endocytosis from the enterocyte brush border during fat absorption

    Hansen, Gert Helge; Niels-Christiansen, Lise-Lotte; Immerdal, Lissi;


    Absorption of dietary fat in the small intestine is accompanied by a rise of intestinal alkaline phosphatase (IAP) in the serum and of secretion of IAP-containing surfactant-like particles from the enterocytes. In the present work, fat absorption was studied in organ cultured mouse intestinal...... clathrin-coated pits. By 60 min, IAP was seen in subapical endosomes and along membranes surrounding fat droplets. IAP is a well-known lipid raft-associated protein, and fat absorption was accompanied by a marked change in the density and morphology of the detergent-resistant membranes harboring IAP...... explants. By immunofluorescence microscopy, fat absorption caused a translocation of IAP from the enterocyte brush border to the interior of the cell, whereas other brush-border enzymes were unaffected. By electron microscopy, the translocation occurred by a rapid (5 min) induction of endocytosis via...

  8. The tillage effect on the soil acid and alkaline phosphatase activity

    Lacramioara Oprica


    Full Text Available Phosphatases (acid and alkaline are important in soils because these extracellular enzymes catalyze the hydrolysis of organic phosphate esters to orthophosphate; thus they form an important link between biologically unavailable and mineral phosphorous. Phosphatase activity is sensitive to environmental perturbations such as organic amendments, tillage, waterlogging, compaction, fertilizer additions and thus it is often used as an environmental indicator of soil quality in riparian ecosystems. The aim of the study was to assess the effect of tillage systems on phosphatases activity in a field experiment carried out in Ezăreni farm. The phosphatase activitiy were determined at two depths (7-10 cm and 15-25cm layers of a chernozem soil submitted to conventional tillage (CT in a fertilised and unfertilised experiment. Monitoring soil alkaline phosphatase activity showed, generally, the same in fertilized soil profiles collected from both depths; the values being extremely close. In unfertilized soils, alkaline phosphatase activity is different only in soils that were exposed to unconventional work using disc harrows and 30cm tillage. Both works type (no tillage and conventional tillage cause an intense alkaline phosphatase activity in 7-10 cm soil profile. Acid phosphatase activity is highly fluctuating in both fertilized as well unfertilized soil, this enzyme being influenced by the performed works.

  9. Expression and Characterization of Recombinant Thermostable Alkaline Phosphatase from a Novel Thermophilic Bacterium Thermus thermophilus XM

    Jianbo LI; Limei XU; Feng YANG


    A gene (tap) encoding a thermostable alkaline phosphatase from the thermophilic bacterium Thermus thermophilus XM was cloned and sequenced. It is 1506 bp long and encodes a protein of 501 amino acid residues with a calculated molecular mass of 54.7 kDa. Comparison of the deduced amino acid sequence with other alkaline phosphatases showed that the regions in the vicinity of the phosphorylation site and metal binding sites are highly conserved. The recombinant thermostable alkaline phosphatase was expressed as a His6-tagged fusion protein in Escherichia coli and its enzymatic properties were characterized after purification. The pH and temperature optima for the recombinant thermostable alkaline phosphatases activity were pH 12 and 75 ℃. As expected, the enzyme displayed high thermostability, retaining more than 50% activity after incubating for 6 h at 80 ℃. Its catalytic function was accelerated in the presence of 0.1 mM Co2+, Fe2+, Mg2+, or Mn2+ but was strongly inhibited by 2.0 mM Fe2+. Under optimal conditions, the Michaelis constant (Km) for cleavage of p-nitrophenyl-phosphate was 0.034 mM. Although it has much in common with other alkaline phosphatases, the recombinant thermostable alkaline phosphatase possesses some unique features, such as high optimal pH and good thermostability.

  10. Alkaline phosphatase expression during relapse after orthodontic tooth movement

    Pinandi Sri Pudyani


    Full Text Available Background: The increasing of osteoblast activities during bone formation will be accompanied with the increasing expression of alkaline phosphatase enzyme (ALP. ALP can be obtained from clear fluid excreted by gingival crevicular fluid (GCF. Bone turnover, especially bone formation process, can be monitored through the expression of ALP secreted by GCF during orthodontic treatment. Thus, retention period is an important period that can be monitored through the level of bone metabolism around teeth. Purpose: This research were aimed to determine the relation of distance change caused by tooth relapse and ALP activities in gingival crevicular fluid after orthodontic; and to determine ALP as a potential biomarker of bone formation during retention period. Methods: Lower incisors of 25 guinea pigs were moved 3 mm to the distally by using open coil spring. Those relapse distance were measured and the gingival crevicular fluid was taken by using paper points to evaluate ALP levels on days 0, 3, 7, 14 and 21 respectivelly by using a spectrophotometer (405 nm. t-test and ANOVA test were conducted to determine the difference of ALP activities among the time intervals. The correlation regression analysis was conducted to determine the relation of distance change caused by the relapse tooth movement and ALP activities. Results: The greatest relapse movement was occurred on day 3 after open coil spring was removed. There was significant difference of the average of distance decrease among groups A1-A5 (p<0.05. It was also known that ALP level was increased on day 3, but there was no significant difference of the average level of ALP among groups A1-A5 (p>0.05. Finally, based on the results of correlation analysis between the ALP level decreasing and the relapse distance on both right and left of mesial and distal sides, it is known that there was no relation between those two variables (p>0.05. Conclusion: It can be concluded that relapse after orthodontic

  11. “补肾健脾”针刺法对绝经后骨质疏松症模型血清碱性磷酸酶水平的影响%Effect of acupuncture method of"invigorating the kidney and strengthening the spleen"on the level of serum alkaline phosphatase in postmenopausal osteoporosis model

    李灿; 黎喜平; 陈久毅; 沈骏; 秧荣昆; 王建


    目的:观察“补肾健脾”针刺法对去卵巢大鼠模型血清中碱性磷酸酶(ALP)含量的影响。方法:将32只雌鼠等分为假手术组、模型组、针刺组及雌激素组。假手术组不摘除卵巢,余3组摘除双侧卵巢,造成绝经后骨质疏松症模型。假手术组及模型组每日同时灌服等量生理盐水,针刺组针刺双侧足三里穴、肾俞穴、脾俞穴、大杼穴及命门穴、关元穴,雌激素组用己烯雌酚溶液灌胃。3个月后取股动脉血,测血清ALP含量。结果:模型组血清碱性磷酸酶含量比假手术组升高(P<0.05);针刺组与雌激素组血清ALP含量较模型组降低(P<0.01)。结论:去卵巢大鼠血清中碱性磷酸酶含量升高,形成高转换骨质疏松症模型,通过雌激素替代和针刺治疗后,血清ALP含量降低,两种治疗均能对抗高转换型骨代谢,为“补肾健脾”治疗绝经后骨质疏松症提供部分临床作用机理。%Objective:To observe the effect of acupuncture method of"invigorating the kidney and strengthening the spleen"on the content of alkaline phosphatase (ALP) in serum of ovariectomized rats.Methods:32 female rats were divided into the sham operation group,the model group,the acupuncture group and the estrogen group.The sham operation group did not remove the ovaries,while bilateral ovaries were removed in the other 3 groups,to made postmenopausal osteoporosis model.The sham operation group and model group were administered with equivalent saline at the same time.The acupuncture group was treated by acupuncture at bilateral Zusanli and Shenyu, Piyu points,Dazhu caveand Mingmen and Guanyuan point,and the estrogen group was treated with intragastric administration of diethylstilbestrol solution.Femoral artery blood was taken after 3 months,and the content of ALP in serum was measured.Results:The serum alkaline phosphatase levels in the model group were higher than those in the sham operation

  12. Radioimmunoassay for prostatic acid phosphatase in human serum. Methodologic aspects

    Pradalier, N.; Canal, P.; Pujol, A.; Fregevu, Y. (Groupe de Recherches du Centre Claudius-Regaud, Toulouse (France)); Soula, G. (Faculte des Sciences Pharmaceutiques, Toulouse (France))


    We propose a double antibody radioimmunoassay for human prostatic acid phosphatase (PAP) in serum for diagnosis and management of prostatic adenocarcinoma under treatment. The antigen is purified from human prostatic fluid by a gel-filtration on Sephadex G 100 followed by affinity chromatography on Con A Sepharose. A specific antibody is raised in rabbits and purified by immunoadsorption with a female serum. The described technique offers both radioisotopic sensibility and immunologic specificity. Physiological values determined in the serum of 125 healthy males are below 2 ng/ml. No significative differences are observed with age. The proposed technique also shows significant differences between values evaluated for benign prostatic hyperplasia and prostatic adenocarcinoma.


    Mauren Picada Emanuelli


    Full Text Available

    The enzymes γ-Glutamyltransferase (GGT and Akaline phosphatase (AP are serum markers of cholestasis process, becoming an important way of diagnosis in hepatic diseases.  Urea and creatinine are eliminated by urine. When these metabolic elements are higher then normal, it is subside to diagnosis mammals’ renal dysfunction. This present study aimed to establish reference values for these enzymes in rabbits (laboratory animals, to obtain data for their use in scientific experiments. Thus, it was collected blood samples in 45 animals; three samples each one, making a total of 135 blood samples. Serum was separated by immediate centrifugation. Colorimetric method was realized and values from 36.44+/-10.66mg/dl for urea, minimum at 9.24mg/dl and a maximum at 66.06mg/dl; 0.94+/-0.22 mg/dl for creatinine, minimum at 0.51mg/dl and a maximum at 1.53mg/dl; and 72.41+/-29.68UI for AP, minimum at 10.66UI and a maximum at 167.39UI, were determinate. The GGT was determined for kinetic method and values from 6.85+/-3.31, minimum at 2UI and a maximum at 15UI.

    KEY WORDS: Hepatic function, rabbits, renal function, serum biochemistry

    As enzimas gama-glutamil transferase e fosfatase alcalina são marcadores séricos de processos colestásicos, sendo importantes no diagn

  14. Acid- and alkaline phosphatase in amniotic fluid in normal and complicated pregnancy.

    Beckman, G; Beckman, L; Löfstrand, T


    171 samples of amniotic fluid were obtained by abdominal amniocentesis from 67 women with complicated pregnancies (isoimmunization, diabetes mellitus or toxaemia). The levels of heat-labile alkaline phosphatase (HLAP), heat-stable alkaline phosphatase (HSAP) and acid phosphatase (AcP) were determined and compared to the enzyme levels in 179 samples from women with normal pregnancies of corresponding gestational ages. HLAP showed two "peaks" of activity, one in the 5th-22nd week and the other at term. HSAP and AcP showed increased activity at term. HSAP was decreased (p less than 0.01) in isoimmunization between the 36th and 40th week. 11 cases of toxaemia with placental insufficiency showed no differences in the levels of HLAP and HSAP compared with normal pregnancy. AcP showed no differences between normal and complicated pregnancy. Samples contaminated by blood showed no significant increase in the acid- and alkaline phosphatase levels. Samples contaminated by meconium showed a complex pattern. Some samples had normal enzyme levels, some had high levels of HLAP only and some had high levels of HSAP and AcP. The origin of the enzymes is not known with certainty. HSAP in amniotic fluid is most likely not of placental but intestinal origin. Determinations of acid- and alkaline phosphatase in amniotic fluid seem to be of little values in the clinical management of complicated pregnancy.

  15. Relation of salivary calcium, phosphorus and alkaline phosphatase with the incidence of dental caries in children

    Vijayaprasad K


    Full Text Available Aim: The purpose of this study was to assess possible relationship of Calcium, Phosphorus and Alkaline-phophatase levels in saliva with incidence of caries in child patients. Settings and Design: Children (n=75 attending Department of Pedodontics, St. Joseph Dental college, Eluru, with and without caries were categorized in to Group I: Consisting of 25 children with non-rampant caries, Group II: Consisting of 25 children with rampant caries, Group III: Consisting of 25 children without caries. (Control group. Materials and Methods: The samples of saliva were collected one week after oral prophylaxis. Unstimulated directly expectorated whole saliva samples were collected in clean, dry, sterilized glass bottles and fitted with proper rubber stoppers immediately. The samples were subjected to biochemical assay for estimation of calcium, phosphorus and alkaline phosphatase levels. Statistical analysis used: ANOVA. Results: The alkaline Phosphatase activity for rampant caries group was 18.66 K.A, and control group was 4.68 K.A. The values of alkaline phosphatase activity for minimal caries group was 6.16 KA. Conclusion: Saliva could reflect a caries risk situation was supported by the fact that alkaline phosphatase activity was very much significantly higher in caries prone groups.

  16. Alkaline phosphatase immobilization onto Bio-Gide(R) and Bio-Oss(R) for periodontal and bone regeneration.

    Oortgiesen, D.A.W.; Plachokova, A.S.; Geenen, C.; Meijer, G.J.; Walboomers, X.F.; Beucken, J.J.J.P van den; Jansen, J.B.M.J.


    AIM: To evaluate the effect of alkaline phosphatase (ALP) immobilization onto Bio-Gide((R)) in vitro, and to study the in vivo performance of ALP-enriched Bio-Gide((R)) and/or Bio-Oss((R)) with the purpose to enhance periodontal regeneration. MATERIALS AND METHODS: Alkaline phosphatase ALP was immob

  17. Differentiating intracellular from extracellular alkaline phosphatase activity in soil by sonication.

    Shuping Qin

    Full Text Available Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio  =  1/8 (w/v and power density  =  15 watt ml(-1], the activity of alkaline phosphomonoesterase (phosphatase in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first plateau of enzyme activity was reached between 60 and 100 s, and a second higher plateau after 300 s. We also found that sonication for 100 s under optimal conditions activated most (about 80% of the alkaline phosphatase that was added to an autoclaved soil, while total bacteria number was not affected. Sonication for 300 s reduced the total bacteria number by three orders of magnitude but had no further effects on enzyme activity. Our results indicate that the first plateau of alkaline phosphatase activity was derived from extracellular enzymes attached to soil particles, and the second plateau to the combination of extracellular and intracellular enzymes after cell lysis. We conclude that our adjusted sonication method may be an alternative to the currently used physiological and chloroform-fumigation methods for differentiating intracellular from extracellular phosphatase activity in soil. Further testing is needed to find out whether this holds for other soil types.

  18. Differentiating intracellular from extracellular alkaline phosphatase activity in soil by sonication.

    Qin, Shuping; Hu, Chunsheng; Oenema, Oene


    Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio  =  1/8 (w/v) and power density  =  15 watt ml(-1)], the activity of alkaline phosphomonoesterase (phosphatase) in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first plateau of enzyme activity was reached between 60 and 100 s, and a second higher plateau after 300 s. We also found that sonication for 100 s under optimal conditions activated most (about 80%) of the alkaline phosphatase that was added to an autoclaved soil, while total bacteria number was not affected. Sonication for 300 s reduced the total bacteria number by three orders of magnitude but had no further effects on enzyme activity. Our results indicate that the first plateau of alkaline phosphatase activity was derived from extracellular enzymes attached to soil particles, and the second plateau to the combination of extracellular and intracellular enzymes after cell lysis. We conclude that our adjusted sonication method may be an alternative to the currently used physiological and chloroform-fumigation methods for differentiating intracellular from extracellular phosphatase activity in soil. Further testing is needed to find out whether this holds for other soil types.

  19. A role for intestinal alkaline phosphatase in the maintenance of local gut immunity

    Chen, K.T.; Malo, M.; Beasley-Topliffe, L.K.; Poelstra, K.; Millan, J.; Mostafa, G.; Alam, S.; Ramasamy, S.; Warren, H.; Hohmann, E.; Hodin, R.A.


    Background and Aims: Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor known to dephosphorylate lipopolysaccharide (LPS); however, the role of IAP in the gut response to luminal bacteria remains undefined. We investigated immune responses of wildtype (WT) and IAP-knockout (IAP-KO

  20. The effect of alkaline phosphatase coated onto titanium alloys on bone responses in rats.

    Schouten, C.; Beucken, J.J.J.P. van den; Jonge, L.T. de; Bronkhorst, E.M.; Meijer, G.J.; Spauwen, P.H.M.; Jansen, J.A.


    The enzyme alkaline phosphatase (ALP) was recently proposed as an implant coating material in order to improve the biological performance of orthopedic and dental implants. The present study evaluated the in vivo bone response to electrosprayed coatings, consisting of ALP, calcium phosphate (CaP) or

  1. Alkaline phosphatase-polyresorcinol complex: characterization and application to seed coating.

    Pilar, María C; Ortega, Natividad; Perez-Mateos, Manuel; Busto, María D


    An alkaline phosphatase (EC from Escherichia coli ATCC27257 was immobilized by copolymerization with resorcinol. The phosphatase-polyresorcinol complex synthesized retained about 74% of the original enzymatic activity. The pH and temperature profile of the immobilized and free enzyme revealed a similar behavior. Kinetic parameters were determined: K(m) and K(i) values were 2.44 and 0.423 mM, respectively, for the phosphatase-polyresorcinol complex and 1.07 and 0.069 mM, respectively, for free phosphatase. The thermal and storage stabilities of the immobilized phosphatase were higher than those of the native one. On addition to soil, free enzyme was completely inactivated in 4 days, whereas the phosphatase-polyresorcinol complex was comparatively stable. Barley seed coated with the immobilized enzyme exhibited higher rhizosphere phosphatase activity. Under pot culture conditions, an increase in the soil inorganic phosphorus was detected when the seed was encapsulated with the phosphatase-polyresorcinol complex, and a positive influence on biomass and inorganic phosphorus concentration of shoot was observed.

  2. Effect of alendronate sodium on serum alkaline phosphatase, osteocalcin and bone mineral density in patients with postmenopausal osteoporosis%阿仑膦酸钠对绝经后骨质疏松症患者骨密度及血清碱性磷酸酶、骨钙素的影响

    白孟海; 白洁; 郑蓉; 陈克明; 刘茗露


    目的:探讨阿仑膦酸钠对绝经后骨质疏松症患者骨密度(BMD)及血清碱性磷酸酶(ALP)、骨钙素(BGP)的影响.方法:62例绝经后骨质疏松症患者随机分为2组,治疗组32例,对照组30例.治疗组口服阿仑膦酸钠、阿法迪三;对照组口服阿法迪三.分别于治疗前、治疗6个月、治疗12个月双能X线骨密度仪测定患者正位腰椎(L2~4)、髋部股骨颈(Neck)、大转子(Troch)和Ward’s三角区BMD,全自动生物分析仪测定血清ALP和BGP含量.结果;治疗组治疗6个月、12个月血清ALP和BGP含量与对照组比较有明显差异(P<0.01).治疗组,治疗12个月后L1_4和髋部(Neck,Troch.Ward's)BMD值与治疗前比较显著增加(P<0.01).治疗组与对照组比较L1-4和全髋部BMD值明显差异(P<0.01).结论:阿仑膦酸钠能有效阻止绝经后骨矿丢失,降低血清ALP含量和提高血清BGP含量.%Objective: To investigate the effect of alendronate sodium on serum alkaline phosphatase (ALP), osleocalein and bone mineral density (BMD) in patienis with postmenopausal osteoporosis. Methods,- A iota! Of 62 postmenopausal women with osteoporosis were randomly divided into 2 groups: 32 treatment patients received alendronate sodium and Alpha D3 treatment, and 30 control patients received Alpha D3 only. Before treatment, 6 and 12 months after treatment, the postmenopausal BMD of anterior - posterior lumbar spines and proximal femur were determined by dual - energy X - ray abaorptiometry, and the serum alkaline phosphatase and osteocalein were detected. Results-. Compared with control group, the serum ALP and osteocalcin levels were significantly decreased 6 and 12 months after treatment (P<0.01). Compared with that before treatment, the BMD value of the lumbar spine and proximal femur wag significantly increased 12 months after treatment (P <0.05 ). There was also obvious difference in BMD level of the lumbar spine and proximal femur 12 months afiei treatment between




    Full Text Available Introduction: Bone marrow contains a population of stem cells capable of differentiating to osteoblast and forming the bone nodule by dexamethasone. Material and Methods: The stromal cells of bone marrow obtained from 4 to 6 weeks old Spruge-Dawely male rats were grown in primary culture for 7 days and subcultured for 18 days. The cells were cultured in either DMEM medium containing 15% fetal calf serum and antibiotics as the controls or the above medium supplemented with osteogenic supplements (OS: include 10 mM Na-beta glycerophosphate (Na-betaGp, 10 nM dexamethasone (Dex and 50 g/ml ascordic acid (AsA as the examined cultures. After 6, 12 and 18 days of grow up in subculture, the cultures were examined for mineralization and alkaline phosphatase (Apase expression. Results: Mesenchymal stem cells (MSCs in examined cultures underwent a dramatic change in cellular morphology and a significat increase in Apase activity by day 12. The deposition of a calcified matrix on the surface of the culture flasks became evident between days 12 and 18. Conclusion: The addition of osteogenic supplements (OS to MSCs cultures induced Apase expression that contributes to cellular differentiation and mineralization of extracellular matrix.

  4. Effect of puerarin combined with estradiol on bone tissue and serum calcium, phosphorus, and alkaline phosphatase in ovariectomized rats%葛根素联合雌二醇对去卵巢大鼠骨组织及血钙、磷和碱性磷酸酶的影响

    李海; 王金花; 黄海玲; 陈建海; 黎飚


    目的:了解小剂量葛根素联合雌二醇对去卵巢大鼠的骨组织、血钙、磷和碱性磷酸酶的影响,为中西医结合治疗绝经后骨质疏松症提供实验依据.方法:5月龄健康雌性大白鼠120只,分成5个实验组(每组24只):①假手术组(sham);②去卵巢模型组(OVX);③葛根索组(Pr),皮下注射葛根素,50 mg/kg,1次/d;④雌二醇组(E2),皮下注射雌二醇200 μg/kg,2次/周;⑤小剂量葛根素+雌二醇组(Pr+ E2),皮下注射雌二醇100 μg/kg,2次/周和葛根素25 mg/kg,1次/d.各实验组在第4、8、12和20周,随机取6只大鼠取股骨切片观察骨组织,采血测量血钙、磷和碱性磷酸酶,数据进行统计学分析.结果:OVX组第4、8、12、20周的骨组织呈骨质疏松病理改变,血钙、磷明显低于sham组(P<0.01),OVX组的第4、8、12、20周血碱性磷酸酶均明显高于sham组(P<0.01).3个治疗组各时间的骨组织和血钙、磷和碱性磷酸酶与sham组无统计学意义(P>0.05).小剂量的葛根素联合雌二醇治疗能使去卵巢大鼠骨组织和血钙、磷和血碱性磷酸酶基本恢复正常(P>0.05),与较大剂量的葛根素组或较大剂量的雌二醇组相比无统计学意义(P>0.05).结论:小剂量的雌二醇与葛根素对去卵巢大鼠的骨质疏松症的治疗效果与单独使用较大剂量的葛根素或较大剂量的雌二醇相比治疗效果相近.%Objective: To understand the effect of puerarin combined with estradiol on bone tissue and serum calcium, phosphorus, and alkaline phosphatase in ovariectomized rats, and provide experimental basis for treating postmenopausal osteoporosis with traditional Chinese medicine combined with Western medicine. Methods: One hundred and twenty 5 - month - old female rats were selected and divided into five experimental groups, 24 rats in each group: sham - operation group, ovariectomized model group, puerarin group (the rats were treated with subcutaneous injection of puerarin

  5. Benomyl inhibits phosphorus transport but not fungal alkaline phosphatase activity in a Glomus–cucumber symbiosis

    Larsen, John; Thingstrup, Ida; Jakobsen, Iver;


    when benomyl was applied to the HC at 10 µg g-1 soil, whereas the uptake of 32P from RHC I roots + hyphae) was reduced only at the highest dose of application to the RHC (100 µ g g-1 soil). In contrast to the marked reduction of benomyl on fungal P transport, the activity of fungal alkaline phosphatase......Short-term effects of benomyl on the arbuscular mycorrhizal fungus Glomus caledonium (Nicol. & Gerd.) Trappe and Gerdeman associated with Cucumis sativus L. were studied by measuring effects on fungal P transport and on fungal alkaline phosphatase activity. Mycorrhizal plants were grown in three...... compartment systems where nylon mesh was used to separate n root-free hyphal compartment (HC) and a root + hyphal compartment(RHC) from The main root compartment (RC). Non-mycorrhizal control plants were grown in similar growth units. After 6 wk benomyl was applied to the plants in three ways: as soil...

  6. Significance of bone specific alkaline phosphatase as a tumor marker in malignant bone tumor

    Kim, Sug Jun; Jeon, Dae Geun; Huh, Kwang [Korea Cancer Center Hospital, Seoul (Korea, Republic of)


    The relationship between total alkaline phosphatase activity and bone forming lesion is a well known fact. But alkaline phosphatase consist mainly of two portion (liver, bone). To clarify the exact activity of bone forming tissue, quantitative measurement of BALP is essential. Two finds of tests were performed for their feasibility as a laboratory test (wheat germ lectin vs electrophoresis). We analyzed 40 bony lesion and got 58 samples. Lectin method was simple, economic, with reliable resproducability. Owing to the small number of test sample, we could not identify the relationship between the disease activity and measured BALP level. Further collection of clinical sample and analysis the pattern of BALP on each clinical settings. (author). 8 refs.

  7. Development of conductometric biosensors based on alkaline phosphatases for the water quality control

    Berezhetskyy, A.


    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devoted to creation and optimization of conductometric biosensor based on alkaline phosphatase active microalgae and sol gel technology, the last chapter described application of the proposed algal biosensor for measurements of heavy metal ions toxicity of waste water, general conclusions stating the progresses achieved in the field of environmental monitoring

  8. Development of conductometric biosensors based on alkaline phosphatases for the water quality control

    Berezhetskyy, A


    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devoted to creation and optimization of conductometric biosensor based on alkaline phosphatase active microalgae and sol gel technology, the last chapter described application of the proposed algal biosensor for measurements of heavy metal ions toxicity of waste water, general conclusions stating the progresses achieved in the field of environmental monitoring

  9. Biochemical and functional properties of mammalian bone alkaline phosphatase isoforms during osteogenesis

    Halling Linder, Cecilia


    The human skeleton is a living and dynamic tissue that constantly is being renewed in a process called bone remodeling. Old bone is resorbed by osteoclasts and new bone is formed by osteoblasts. Bone is a composite material made up by mineral crystals in the form of hydroxyapatite (calcium and phosphate) that provides the hardness of bone, and collagen fibrils that provides elasticity and flexibility. Alkaline phosphatase (ALP) is a family of enzymes that is present in most species and cataly...

  10. [Development of conductometric biosensor based on alkaline phosphatase for determining concentration of cadmium ions].

    Sosovs'ka, O F; Berezhets'kyĭ, A L


    The paper describes a novel conductometric biosensor sensitive to cadmium ions based on alkaline phosphatase immobilized on gold planar microelectrodes used as transducers. Assays have been carried out with paranitrophenyl phosphate as substrate for the immobilized enzyme. Various parameters such as reticulation time, along with pH, ionic strength and buffer concentration of the measuring solution were studied. The optimized biosensor was stable, reproducible and it exhibited a detection limit of 4.45 microM for cadmium ions.

  11. Development of conductometric biosensors based on alkaline phosphatases for the water quality control


    Researches are focused on the elaboration of enzymatic microconductometric device for heavy metal ions detection in water solutions. The manuscript includes a general introduction, the first chapter contains bibliographic review, the second chapter described the fundamentals of conductometric transducers, the third chapter examining the possibility to create and to optimize conductometric biosensor based on bovine alkaline phosphatase for heavy metals ions detection, the fourth chapter devote...

  12. Intraspecific variation in alkaline phosphatase activity in Phaeodactylum tricornutum (Bacillariophyceae, Bohlin

    Domênica Teixeira de Lima


    Full Text Available ABSTRACT To describe potential intraspecific variation in phosphorus incorporation in two strains of Phaeodactylum tricornutum (Bohlin, Ub3 and Ub7, alkaline phosphatase (AP activity was evaluated via enzyme-labeled fluorescence assay. Analysis using the probe ELF-97(r provides individual evaluation, and therefore can determine the nutritional status of inorganic phosphorus in phytoplanktonic cells. Bioassays compared the control treatment to both phosphate-enriched and phosphate-depleted treatments by varying only the phosphate concentration in the media. The P. tricornutum strains exhibited differences in their development when incubated in the phosphate-enriched media. The development of the Ub7 strain differed by exhibiting "luxury uptake" and utilization of organic phosphorus, and the alkaline phosphatase analysis indicated limitations of this clone under such conditions. The Ub7 strain showed higher AP activity, when compared to Ub3, in the P-enriched condition. P. tricornutum presented increases in AP activity and low variation in Surface/Volume ratio, by increasing biovolume and its maximum linear dimension, as strategies for phosphate incorporation. Our results highlight intraspecific differences in alkaline phosphatase activity, and hence differences in the incorporation of organic phosphorus, as the tested species regulated enzymatic activity under different external phosphate concentrations.

  13. Synthesis, alkaline phosphatase inhibition studies and molecular docking of novel derivatives of 4-quinolones.

    Miliutina, Mariia; Ejaz, Syeda Abida; Khan, Shafi Ullah; Iaroshenko, Viktor O; Villinger, Alexander; Iqbal, Jamshed; Langer, Peter


    New and convenient methods for the functionalization of the 4-quinolone scaffold at positions C-1, C-3 and C-6 were developed. The 4-quinolone derivatives were evaluated for their inhibitory potential on alkaline phosphatase isozymes. Most of the compounds exhibit excellent inhibitory activity and moderate selectivity. The IC50 values on tissue non-specific alkaline phosphatase (TNAP) were in the range of 1.34 ± 0.11 to 44.80 ± 2.34 μM, while the values on intestinal alkaline phosphatase (IAP) were in the range of 1.06 ± 0.32 to 192.10 ± 3.78 μM. The most active derivative exhibits a potent inhibition on IAP with a ≈14 fold higher selectivity as compared to TNAP. Furthermore, molecular docking calculations were performed for the most potent inhibitors to show their binding interactions within the active site of the respective enzymes.

  14. Alkaline phosphatase inhibition based conductometric biosensor for phosphate estimation in biological fluids.

    Upadhyay, Lata Sheo Bachan; Verma, Nishant


    Determination of phosphate ions concentration is very important from both, environmental and clinical point of view. In this study, a simple and novel conductometric biosensor for indirect determination of the phosphate ions in aqueous solution has been developed. The developed biosensor is based on the inhibition of immobilized alkaline phosphatase activity, in the presence of the phosphate ions. This is the first time we developed a mono-enzymatic biosensor for indirect estimation of phosphate ions. The developed biosensor showed a broad linear response (as compared to other reported biosensors) for phosphate ions in the range of 0.5-5.0 mM (correlation coefficient=0.995), with a detection limit of 50 µM. Different optimized parameters were obtained as the buffer concentration of 30 mM, substrate concentration of 1.0mM, and a pH of 9.0. All the optimized parameters were analyzed by analysis of variance, and were found to be statistically significant at a level of α=0.05. The developed biosensor is also suitable to determine the serum phosphate concentration, with a recovery of 86-104%, while a recovery of 102% was obtained from the water samples that were spiked with 500 µM phosphate. A relative standard deviation in the conductance response for five successive measurements (n=5) did not exceed 7%, with a shelf life of 30 days. With a lower detection limit and a higher recovery, the biosensor provides a facile approach for phosphate estimation in biological fluids.

  15. Copper sulfide nanoparticle-decorated graphene as a catalytic amplification platform for electrochemical detection of alkaline phosphatase activity.

    Peng, Juan; Han, Xiao-Xia; Zhang, Qing-Chun; Yao, Hui-Qin; Gao, Zuo-Ning


    Copper sulfide nanoparticle-decorated graphene sheet (CuS/GR) was successfully synthesized and used as a signal amplification platform for electrochemical detection of alkaline phosphatase activity. First, CuS/GR was prepared through a microwave-assisted hydrothermal approach. The CuS/GR nanocomposites exhibited excellent electrocatalytic activity toward the oxidation of ALP hydrolyzed products such as 1-naphthol, which produced a current response. Thus, a catalytic amplification platform based on CuS/GR nanocomposite for electrochemical detection of ALP activity was designed using 1-naphthyl phosphate as a model substrate. The current response increased linearly with ALP concentration from 0.1 to 100 U L(-1) with a detection limit of 0.02 U L(-1). The assay was applied to estimate ALP activity in human serum samples with satisfactory results. This strategy may find widespread and promising applications in other sensing systems that involves ALP.

  16. Host plant effects on alkaline phosphatase activity in the whiteflies, Bemisia tabaci Biotype B and Trialeurodes vaporariorum.

    Yan, Ying; Peng, Lu; Liu, Wan-Xue; Wan, Fang-Hao; Harris, Marvin K


    Bemisia tabaci (Gennadius) B-biotype and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) often coexist on greenhouse-grown vegetable crops in northern China. The recent spread of B. tabaci B-biotype has largely replaced T. vaporariorum, and B-biotype now overlaps with T. vaporariorum where common hosts occur in most invaded areas. The impact of the B-biotype on the agro eco system appears to be widespread, and involves the ability to compete with and perhaps replace other phytophages like T. vaporariorum. An emerging hypothesis is that the B-biotype is physiologically superior due at least in part to an improved ability to metabolically utilize the alkaline phosphatase pathway. To test this hypothesis, alkaline phosphatase activity was studied in the B-biotype and T. vaporariorum after feeding on a number of different hosts for a range of durations, with and without host switching. Alkaline phosphatase activity in T. vaporariorum was 1.45 to 2.53-fold higher than that of the B-biotype when fed on tomato for 4 and 24 h, or switched from tomato to cotton and cabbage for the same durations. However, alkaline phosphatase activity in the B-biotype was 1.40 to 3.35-fold higher than that of T. vaporariorum when the host switching time was ∼72 and ∼120 h on the same plant. Both short-term (4 h) and long-term (72 h) switching of plant hosts can significantly affect the alkaline phosphatase activity in the two species. After ∼120 h, feeding on tomato and cotton alkaline phosphatase activity in the B-biotype was significantly higher than that of T. vaporariorum. It was shown that alkaline phosphatase aids the species feeding on different plant species, and that the B-biotype is physiologically superior to T. vaporariorum in utilizing the enzyme compared to T. vaporariorum over longer periods of feeding.

  17. Effect of Diazinon on Acid and Alkaline Phosphatase Activities in Plasma and Organs of Clarias gariepinus

    I.R. Inyang; E.R. Daka and E.N. Ogamba


    The aim of this study was to determine the effect of the pesticide, diazinon, on phosphatases in the plasma and organs on Clarias gariepinus. Adult Clarias gariepinus were exposed in four replicates to varying sublethal concentrations diazinon (ranging from 1.00 to 10.0 mg/L) in 30-day semi-static bioassays. Alkaline phoshatase (ALP) and acid phosphate (ACP) were determined in plasma and other organs (gastrointestinal tract - GIT, kidney, muscle, gill and liver) of the fish after the experime...

  18. [Effect of dental alloys on salivary alkaline and acid phosphatase, alpha amylase K+, Na+, and Cl-].

    Todorov, I; Saprjanova, M


    Comparative studied were performed in healthy subjects without metals in their oral cavities and in individuals having different metal alloys (gold, steel, amalgam) in their mouths and presenting with various complaints such as xerostomia, burning mucosa, etc. It was found that the contents of alkaline and acid phosphatases, alpha-amylase, K+, Na+ and Cl- in saliva increased significantly with the increase in total corrosion potential when non-precious metal alloys, especially different types of alloys, were present. Parallel to this, the frequency and the intensity of the complaints increased.

  19. Intestinal alkaline phosphatase: a summary of its role in clinical disease.

    Fawley, Jason; Gourlay, David M


    Over the past few years, there is increasing evidence implicating a novel role for Intestinal Alkaline Phosphatase (IAP) in mitigating inflammatory mediated disorders. IAP is an endogenous protein expressed by the intestinal epithelium that is believed to play a vital role in maintaining gut homeostasis. Loss of IAP expression or function is associated with increased intestinal inflammation, dysbiosis, bacterial translocation and subsequently systemic inflammation. As these events are a cornerstone of the pathophysiology of many diseases relevant to surgeons, we sought to review recent research in both animal and humans on IAP's physiologic function, mechanisms of action and current research in specific surgical diseases.

  20. Comparison of Inactivation and Unfolding of Calf Intestinal Alkaline Phosphatase in Guanidinium Chloride Solution

    张英侠; 闫淑莲; 刘永利; 席宏伟; 周海梦


    The changes in activity and unfolding of calf intestinal alkaline phosphatase (CIP) during denaturation in guanidinium chloride solutions of different concentrations were investigated using ultraviolet difference absorption spectra and fluorescence emission spectra. Unfolding and inactivation rate constants were measured and compared. The inactivation course is much faster than that of unfolding, which suggests that the active site of CIP containing two zinc ions and one magnesium ion is situated in a limited and flexible region of the enzyme molecule, which is more fragile to the denaturant than the protein as a whole.

  1. A New Voltammetric Enzyme Immunoassay System for the Detection of Alkaline Phosphatase

    KuiJIAO; WeiSUN; 等


    A new voltammetric enzyme immunoassay system was invesigated based on p-nitrophenyl phosphate (PNPP) as the subsrate for alkaline phosphatase (ALP). PNPP is enzymatically hydrolyzed and the product p-nitrophenol (PNP) is detected by differential pulse voltammetry (DPV), which can be oxidized at +1.02 V(vs.Ag/AgCl) on bare glass carbon electrode (GCE). The conditions for enzymatic reaction and electrochemical detection were studied. According to this method,ALP can be detected with a detection limit of 2.8×102 mU/L and a linear range of 4.0×102-1.0×106mU/L.

  2. A New Voltammetric Enzyme Immunoassay System for the Detection of Alkaline Phosphatase


    A new voltammetric enzyme immunoassay system was investigated based on p-nitrophenyl phosphate (PNPP) as the substrate for alkaline phosphatase (ALP). PNPP is enzymatically hydrolyzed and the product p-nitrophenol (PNP) is detected by differential pulse voltammetry (DPV), which can be oxidized at +1.02 V (vs. Ag/AgCl) on bare glass carbon electrode (GCE). The conditions for enzymatic reaction and electrochemical detection were studied. According to this method, ALP can be detected with a detection limit of 2.8′102 mU/L and a linear range of 4.0′102 ~ 1.0′106 mU/L.

  3. [Behavior of serum alkaline during pregnancy. II. Pathological pregnancy].

    Stark, K H; Nabel, H J; Kyank, H; Neumayer, E; Dässler, C G; Töwe, J


    832 estimations of heat stable alkaline phosphatase (HSAP) and of heat alkaline phosphatase (HLAP) were carried out simultaneously in late pregnant women at 25th to the 42nd weeks of pregnancy. 147 of them delivered children with normal birth-weight. All these women suffered from pre-eclampsia, hypertension or any kind of superimposed pre-eclampsia. 110 other pregnant women with or without symptoms of pre-eclampsia gave birth to small for dates babies. In addition, the values of these patients were compared with 372 estimations of the same enzymes carried out in 120 patients with normal pregnancy and outcome of normal weighted children. The site of the values of every group showed no typical correlation to the course and outcome of their pregnancy. Regarding four special criterions it was possible to give a good prediction by serial determinations for the weight of the newborn in 80 per cent of the cases. A correlation between the urinary excretion of total oestrogens as well as HLAP and the values of HSAP was to be found only in some groups of patients.

  4. Changes in plasma inorganic phosphorus and alkaline phosphatase activity during the adolescent growth spurt.

    Round, J M; Butcher, S; Steele, R


    Longitudinal data on changes in the concentrations of plasma inorganic phosphorus and plasma alkaline phosphatase activity in 23 girls and 44 boys during the adolescent growth spurt are reported together with the height velocities, ages and sexual maturity ratings. The average age at the peak of the growth spurt was 12.5 years in the girls and 14.1 years in the boys with mean annual height gains of 7.0 and 9.7 cm/year respectively. In both sexes, plasma alkaline phosphatase activity rose and fell with the growth velocity during the growth spurt. Plasma inorganic phosphorus rose to reach a peak in the 4 months before the peak of the growth spurt in height; this rise was statisically significant in the boys but not in the girls. Values subsequently fell rapidly towards the normal adult concentrations. Plasma calcium, total protein, and albumin concentrations were also followed during this time, but were not at any point significantly different from normal adult values. These findings provide a guide for the interpretation of plasma biochemistry in adolescent patients.

  5. Alkaline phosphatase activity and the phosphorus mineralization rate of Lake Taihu

    GAO; Guang; ZHU; Guangwei; QIN; Boqiang; CHEN; Jun


    The phosphorus fractions, the alkaline phosphatase activity (APA) and other water chemical parameters were concomitantly monitored from April 2003 to October 2004 in different ecotype sites of Lake Taihu. During the stages of algae growth, the phosphorus fractions and their relationships with APA in different ecotype sites were discussed and the phosphorus mineralization rate was calculated. In the water of Lake Taihu, most of the phosphorus (70.2%) could be attributed to the suspended particulate phosphorus, while the dissolved reactive phosphorus (DRP) seems to contribute less than 7%. About 58% of the total phosphorus, however, can be hydrolyzed as inorganic phosphate to compensate for phosphorus deficiency of algae and bacteria growth. During the different algae growth stages, the APA and its Kinetic parameters were varied significantly between different ecotype sites of Lake Taihu. This trend is also visible by comparing the phosphorus mineralization rate,and the most rapidly phosphorus turnover time is only several minutes. The fast recycle of phosphorus can, to some extent, be explained that the phosphorus source of algal blooms. The phytoplankton seems to compensate for phosphorus deficiency by using the alkaline phosphatase to hydrolyze phosphomonoesters.

  6. Immobilization of Penaeus merguiensis alkaline phosphatase on gold nanorods for heavy metal detection.

    Homaei, Ahmad


    Biotechnology of enzyme has gained popularity due to the growing need for novel environmental technologies and the development of innovative mass-production. The work describes the original application of biosensors based on Penaeus merguiensis alkaline phosphatase (PM ALP) immobilized on gold nanorods (GNRs) to heavy metal determination. Penaeus merguiensis alkaline phosphatase (PM ALP) was immobilized on gold nanorods (GNRs) by ionic exchange and hydrophobic interactions. The optimum pH and temperature for maximum enzyme activity for the immobilized PM ALP are identified to be 11.0 and 60°C, respectively, for the hydrolysis of para-Nitrophenylphosphate (p-NPP). The kinetic studies confirm the Michaelis-Menten behavior and suggests overall slightly decrease in the performance of the immobilized enzyme with reference to the free enzyme. Km and Vmax values were 0.32µm and 54µm. min(-1) for free and 0.39µm and 48µmmin(-1) for immobilized enzymes, respectively. Similarly, the thermal stability, storage stability and stability at extreme pH of the enzyme is found to increase after the immobilization. The inhibitory effect heavy metal ions was studied on free and immobilized PM ALP. The bi-enzymatic biosensor were tested to study the influence of heavy metal ions and pesticides on the corresponding enzyme. The obtained high stability and lower decrease in catalytic efficiency suggested the great potential and feasibility of immobilized PM ALP nanobiocatalyst in efficient and apply the biosensor in total toxic metal content determination.

  7. Ultrastructural localization of alkaline phosphatase in the eggs of Hydatigera taeniaeformis (Taenia taeniaeformis).

    Ajayi, S T; Smith, B F; LeFlore, W B


    Freshly shed gravid proglottids from a three-month-old infection of Hydatigera taeniaeformis collected from the faecal droppings of infected cats were used for this study. They were treated for transmission electron microscopy (TEM) followed by incubation using the lead precipitate method. Control sections were incubated in a substrate-free medium, a substrate medium containing 1.0 mM sodium fluoride (NaF) (an inhibitor), and the last sections were denatured at 90 degrees C for 1 min prior to incubation. Intensive alkaline phosphatase activity in the embryophoric blocks and the outer embryophoric membrane was revealed. The reaction products were also indicated in the oncospheral membrane. However, no enzyme activity was seen in any other part of the egg. The enzyme was also absent in the control sections. The presence of alkaline phosphatase activity in the outer embryophoric and oncospheral membranes suggested that this enzyme may be involved in carbohydrate metabolism and nutritional absorption, and also may play a role in the transport of nutrients and other substances from the adult to the developing embryo, respectively.

  8. Study on the activity of serum bone alkaline phosphatase and its relations to the heritability among prepuberty Twins%青春前期双生子血清骨碱性磷酸酶活性测定及遗传度分析

    王文军; 张璟; 李晶; 刘琥; 孔庆胜; 周军燕; 随桂英; 邓磊


    目的 通过对青春前期双生子血清骨碱性磷酸酶(bone alkaline phosphatase,BALP)活性的测定,分析血清BALP活性的遗传度,评价特定人群钙的缺乏状况和机体对钙的需求情况.方法 调查9~16岁双生子73对,利用骨源性碱性磷酸酶试剂盒进行血清BALP活性测定.在DNA卵型鉴定基础上.以组内相关系数法及Christian遗传度计算公式分析血清BALP活性的遗传度. 结果 经卵型鉴定,73对双生子中同卵双生子34对,异卵双生子39对;BALP>250 U/L的人占43.1%,BALP为200~250 U/L占54.8%,BALP≤200 U/L占2.1%;男性中钙摄入不足者占48.4%,女性中占39.0%;各年龄组钙的摄入能满足机体需求的均不到10.0%;尤其是10~13岁年龄段儿童,明确缺钙者所占比例均>45.0%;不同性别、不同年龄间BALP均值差异无统计学意义,性别t=1.633,P=0.105;年龄F=0.323,P=0.924.经过遗传度分析,单卵双生(MZ)对内方差=191.54,对间方差=1462.22,相关系数=0.77;双卵双生(DZ)对内方差=491.03,对间方差=1475.57,相关系数=0.50;BALP活性的遗传度为0.54. 结论 青春前期的双生子普遍存在缺钙问题;BALP的活性遗传因素占54%,环境因素占46%.%Objective To evaluate calcium deficiency and demand in pre-puberty twins and to analyze the heritability of serum bone alkaline phosphatase(BALP).Methods A total of 73 pairs of twins aged 9-16 years were examined by BALP test.Microsatellite polymorphism Was used To diagnose the zygosity of twins,while both intraclass correlation coefficient method and Christian formula were perfortled to investigate heritability of serum BALP.Results The results of zygosity diagnosis displayed that 34 pairs of twins were monozygotic(MZ)twins and 39 pairs were dizygotic(DZ)twins.97.9%of the subjects appeared unusual in the activity of BALP,with activity of BALP>250 U/L in 43.1%of subjects and 200-250 U/L in 54.8%of the subjects.The intake of calcium Was unsatisfied in 48.4%of the boys and 39

  9. Serum alkaline phosphatases concentration in adult patients with skeleton class Ⅱ malocclusion:preliminary results%成人骨性Ⅱ类错牙合畸形患者血清碱性磷酸酶浓度的初步研究

    马艳; 施优灵; 李金超; 刘继光


    Objective To explore the concentration and roles of serum alkaline phosphatases ( ALP) in patients with skeleton class Ⅱ malocclusion. Methods 45 adult patients with skeleton class Ⅱ malocclusion and 47 age-and sex-matched healthy controls were se-lected in this study, and the concentrations of serum ALP were collected. The difference between experimental and control groups was assessed by independent Student's t-test with SPSS 18. 0 software. Results The serum ALP level in patients and that in healthy con-trols were (41. 62±1. 97)U/L and (65. 83±2. 70)U/L, respectively. The serum ALP level in male patients and that in healthy con-trols were (52.07±4.63)U/L and (68.25±2.65)U/L, whereas that in female patients and that in healthy controls were (36.40± 0. 92)U/L and (64. 04±4. 28) U/L, respectively. The serum ALP level in patients25 years old vs. that in healthy controls was (41. 20±2. 45)U/L vs. (66. 00±3. 35)U/L, and (51. 29±18. 68)U/L vs. (66. 88±11. 76)U/L for males, while (37. 28±5. 56)U/L vs (65. 18±24. 68)U/L for females. The significant statistical difference was observed (P<0. 05). Conclusion The serum ALP level in adult patients with skeleton class Ⅱ malocclusion is lower than that in healthy controls, suggesting weaker bone metabolism in the patients, which may be resulted from mandibular hypodevelopment.%目的 研究骨性Ⅱ类错牙合畸形患者的血清碱性磷酸酶浓度及意义.方法 选择45例成人骨性Ⅱ类错牙合畸形患者和47例年龄和性别匹配的正常对照成人,检测其血清碱性磷酸酶浓度.应用SPSS 18.0软件对检测值进行成组t检验统计学分析.结果 成人骨性Ⅱ类错牙合畸形患者和正常对照成人的血清ALP浓度分别为(41.62±1.97)U/L和(65.83±2.70)U/L.其中男性骨性Ⅱ类错牙合畸形患者和对照成人的血清ALP浓度分别为(52.07±4.63)U/L和(68.25±2.65)U/L,女性分别为(36.40±0.92)U/L和(64.04±4.28)U/L.25岁以下骨性Ⅱ类错牙合畸形患

  10. Transferability studies for the AACC reference method and the IFCC method for measurement of alkaline phosphatase activity.

    Tietz, N W; Rinker, A D; Burtis, C; Duncan, P; Ervin, K; Ewen, L; Hørder, M; Mathieu, M; Petitclerc, C; Grisley, D


    We present the results of measurements of alkaline phosphatase activity from interlaboratory transferability studies conducted by 12 laboratories in five countries. The variability, as demonstrated by within-day precision (CV less than or equal to 2.6%), between-day precision (CV less than or equal to 3.6%), and between-lab precision (CV less than or equal to 6.3%) establishes the transferability of this method for alkaline phosphatase. Some common errors encountered in enzyme measurements that affect the accuracy and precision of the measurements are listed.

  11. Purification and characterization of an alkaline phosphatase induced by phosphorus starvation in common bean (Phaseolus vulgaris L.) roots

    Morales, L.; Gutierrez, N.; Maya, V.; Parra, C.; Martinez B, E.; Coello, P., E-mail: [UNAM, Facultad de Quimica, Departamento de Bioquimica, Ciudad Universitaria, 04510 Mexico D. F. (Mexico)


    Two phosphatase isoforms from roots of the common bean (Phaseolus vulgaris L.) showed an increase in activity in response to phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass of APIII was 35 kDa by both SDS-Page and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of ph 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phospho enol-pyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyro phosphatase activity of this enzyme, the hydrolysis of pyro phosphatase increased substantially in the presence of Mg{sup 2+}.

  12. A Disposable Alkaline Phosphatase-Based Biosensor for Vanadium Chronoamperometric Determination

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia


    A chronoamperometric method for vanadium ion determination, based on the inhibition of the enzyme alkaline phosphatase, is reported. Screen-printed carbon electrodes modified with gold nanoparticles were used as transducers for the immobilization of the enzyme. The enzymatic activity over 4-nitrophenyl phosphate sodium salt is affected by vanadium ions, which results in a decrease in the chronoamperometric current registered. The developed method has a detection limit of 0.39 ± 0.06 μM, a repeatability of 7.7% (n = 4) and a reproducibility of 8% (n = 3). A study of the possible interferences shows that the presence of Mo(VI), Cr(III), Ca(II) and W(VI), may affect vanadium determination at concentration higher than 1.0 mM. The method was successfully applied to the determination of vanadium in spiked tap water. PMID:24569772

  13. A Disposable Alkaline Phosphatase-Based Biosensor for Vanadium Chronoamperometric Determination

    Ana Lorena Alvarado-Gámez


    Full Text Available A chronoamperometric method for vanadium ion determination, based on the inhibition of the enzyme alkaline phosphatase, is reported. Screen-printed carbon electrodes modified with gold nanoparticles were used as transducers for the immobilization of the enzyme. The enzymatic activity over 4-nitrophenyl phosphate sodium salt is affected by vanadium ions, which results in a decrease in the chronoamperometric current registered. The developed method has a detection limit of 0.39 ± 0.06 µM, a repeatability of 7.7% (n = 4 and a reproducibility of 8% (n = 3. A study of the possible interferences shows that the presence of Mo(VI, Cr(III, Ca(II and W(VI, may affect vanadium determination at concentration higher than 1.0 mM. The method was successfully applied to the determination of vanadium in spiked tap water.


    周正; 周坚; 邹石莹; 吴效民


    Objective. To discover the relation between alkaline phosphatase (ALP) in gingival crevicular fluid (GCF)of implant teeth aad the curing results.Methods. We measured the ALP level in GCF among 56 cases of implant teeth which included 2 failed cas-es, 5 cases with bad oral hygiene and gingivitis, and compared it with that in the normal group composed of 10persons.Results. The ALP levels in normal group and success implant group showed no difference. The ALP levelsin normal group and success with gingivitis group showed obvious difference. The ALP levels of the 2 failed cas-es are the highest of all.Conclusions. The ALP level in GCF is an important index in evaluating the curing result of the implantteeth.``

  15. Relationship between Salivary Alkaline Phosphatase Enzyme Activity and The Concentrations of Salivary Calcium and Phosphate Ions

    Mina Jazaeri


    Full Text Available Although salivary alkaline phosphatase (ALP can balance de- and remineralization processes of enamel, there is no evidence regarding its effects on the concentrations of calcium and phosphate in saliva. The present study aims to determine the relationship between salivary ALP activity and the concentrations of calcium and phosphate in saliva. In this cross-sectional study, we evaluated salivary markers in 120 males, ages 19 to 44 years. All participants provided 5 mL of unstimulated whole saliva and the level of enzyme activity as well as calcium and phosphate concentrations were measured using a colorimetric method. Data were gathered and analyzed by statistical package for social sciences (SPSS 13.00 using Pearson correlation test. A p value of 0.05. According to the results of the present study, there was no significant relation between salivary ALP activity and calcium and phosphate concentrations in saliva. However, further research is highly recommended.

  16. Improvement of Student Understanding of How Kinetic Data Facilitates the Determination of Amino Acid Catalytic Function through an Alkaline Phosphatase Structure/Mechanism Bioinformatics Exercise

    Grunwald, Sandra K.; Krueger, Katherine J.


    Laboratory exercises, which utilize alkaline phosphatase as a model enzyme, have been developed and used extensively in undergraduate biochemistry courses to illustrate enzyme steady-state kinetics. A bioinformatics laboratory exercise for the biochemistry laboratory, which complements the traditional alkaline phosphatase kinetics exercise, was…

  17. Multiple unfolding intermediates of human placental alkaline phosphatase in equilibrium urea denaturation.

    Hung, H C; Chang, G G


    Alkaline phosphatase is an enzyme with a typical alpha/beta hydrolase fold. The conformational stability of the human placental alkaline phosphatase was examined with the chemical denaturant urea. The red shifts of fluorescence spectra show a complex unfolding process involving multiple equilibrium intermediates indicating differential stability of the subdomains of the enzyme. None of these unfolding intermediates were observed in the presence of 83 mM NaCl, indicating the importance of ionic interactions in the stabilization of the unfolding intermediates. Guanidinium chloride, on the other hand, could stabilize one of the unfolding intermediates, which is not a salt effect. Some of the unfolding intermediates were also observed in circular dichroism spectroscopy, which clearly indicates steady loss of helical structure during unfolding, but very little change was observed for the beta strand content until the late stage of the unfolding process. The enzyme does not lose its phosphate-binding ability after substantial tertiary structure changes, suggesting that the substrate-binding region is more resistant to chemical denaturant than the other structural domains. Global analysis of the fluorescence spectral change demonstrated the following folding-unfolding process of the enzyme: N I(1) I(2) I(3) I(4) I(5) D. These discrete intermediates are stable at urea concentrations of 2.6, 4.1, 4.7, 5.5, 6.6, and 7.7 M, respectively. These intermediates are further characterized by acrylamide and/or potassium iodide quenching of the intrinsic fluorescence of the enzyme and by the hydrophobic probes, 1-anilinonaphthalene-8-sulfonic acid and 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid. The stepwise unfolding process was interpreted by the folding energy landscape in terms of the unique structure of the enzyme. The rigid central beta-strand domain is surrounded by the peripheral alpha-helical and coil structures, which are marginally stable toward a chemical

  18. Comparative Analysis of Salivary Alkaline Phosphatase in Post menopausal Women with and without Periodontitis

    Sophia, Khumukcham; Sudhakar, Uma; Jayakumar, Parvathee; Mathew, Danny


    Introduction Alkaline phosphatase is an intracellular destruction enzyme in the periodontium, and it takes part in the normal turnover of the periodontal ligament, alveolar bone, and root cementum formation and maintenance. Aim The aim of this case control study was to evaluate the enzyme Alkaline Phosphatase (ALP) level in saliva of post menopausal women with and without chronic periodontitis. Materials and Methods In this study, 40 individuals, satisfying the study inclusion and exclusion criteria, were recruited. They were categorically divided, on the basis of gingival index, probing pocket depth and clinical attachment level, into two groups: Group I (post menopausal women with a clinically healthy periodontium, n=20); and Group II (post menopausal women with generalized chronic periodontitis, n=20). Clinical parameters assessed were Plaque Index (PI), Gingival Index (GI), Clinical Attachment Level (CAL) and Probing Pocket Depth (PPD). Unstimulated salivary samples were obtained in which the ALP concentration was measured using p-Nitrophenylphosphate, and 2-amino-2-methyl-1-propanol reagents in Beckman and Coulter, AU 480 auto analyser. Mann-Whitney U test was used to find statistical difference with respect to all clinical parameters such as PI, GI, CAL, PPD and salivary ALP levels. Results The mean ALP in saliva was found to be higher in Group II compared to Group I and the difference was statistically significant with the p-value of 0.008. Conclusion A noteworthy increase in the ALP concentration was seen in saliva in our study (Group II) may be due to increased periodontal inflammation in post menopausal women. Thus salivary ALP can be taken as an additional biomarker to early diagnosis, development and progression of periodontitis especially among post menopausal women. PMID:28274061

  19. Zein as biodegradable material for effective delivery of alkaline phosphatase and substrates in biokits and biosensors.

    Jornet-Martínez, N; Campíns-Falcó, P; Hall, E A H


    A biodegradable material, zein, is proposed as a reagent delivery platform for biokits and biosensors based on alkaline phosphatase (ALP) activity/inhibition in the presence of phosphatase substrates. The immobilization and release of both the substrate and/or the active ALP, in a biodegradable and low-cost material such as zein, a prolamin from maize, and in combination with glycerol as plasticizer have been investigated. Three zein-based devices are proposed for several applications: (1) inorganic phosphorus estimation in water of different sources (river, lake, coastal water and tap water) with a detection limit of 0.2mg/L - compared to at least 1mg/L required by legislation, (2) estimation of ALP in saliva and (3) chlorpyrifos control in commercial preparations. The single-use kits developed are low cost, easy and fast to manufacture and are stable for at least 20 days at -20°C, so the zein film can preserve and deliver both the enzyme and substrates.

  20. Electrochemical Detection of Alkaline Phosphatase in BALB/c Mouse Fetal Liver Stromal Cells with Capillary Electrophoresis

    Xue Mei SUN; Dong LI; Zeng Liang BAI; Wen Rui JIN


    A method for determination of alkaline phosphatase (ALP) in BALB/c mouse fetal liver stromal cells has been described based on the catalytic reaction. After the cell extract is incubated with the substrate disodium phenyl phosphate, the reaction product phenol generated by ALP is determined by capillary electrophoresis with electrochemical detection.

  1. Changes in Bone Alkaline Phosphatase and Procollagen Type-1 C-Peptide after Static and Dynamic Exercises

    Kubo, Keitaro; Yuki, Kazuhito; Ikebukuro, Toshihiro


    We investigated the effects of two types of nonweight-bearing exercise on changes in bone-specific alkaline phosphatase (BAP) and pro-collagen type 1 C-peptide (P1P). BAP is a specific marker of bone synthesis, whereas P1P reflects synthesis of type 1 collagen in other organs as well as bone. Eight participants performed static and dynamic…

  2. Structural characteristics of alkaline phosphatase from the moderately halophilic bacterium Halomonas sp. 593

    Arai, Shigeki; Yonezawa, Yasushi [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan); Ishibashi, Matsujiro [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Matsumoto, Fumiko; Adachi, Motoyasu; Tamada, Taro [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan); Tokunaga, Hiroko [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Blaber, Michael [Florida State University, 1115 West Call Street, Tallahassee, FL 32306-4300 (United States); Tokunaga, Masao [Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065 (Japan); Kuroki, Ryota, E-mail: [Japan Atomic Energy Agency, 2-4 Shirakata-shirane, Tokai, Ibaraki 319-1195 (Japan)


    In order to clarify the structural basis of the halophilic characteristics of an alkaline phosphatase derived from the moderate halophile Halomonas sp. 593 (HaAP), the tertiary structure of HaAP was determined to 2.1 Å resolution by X-ray crystallography. The structural properties of surface negative charge and core hydrophobicity were shown to be intermediate between those characteristic of halophiles and non-halophiles, and may explain the unique functional adaptation to a wide range of salt concentrations. Alkaline phosphatase (AP) from the moderate halophilic bacterium Halomonas sp. 593 (HaAP) catalyzes the hydrolysis of phosphomonoesters over a wide salt-concentration range (1–4 M NaCl). In order to clarify the structural basis of its halophilic characteristics and its wide-range adaptation to salt concentration, the tertiary structure of HaAP was determined by X-ray crystallography to 2.1 Å resolution. The unit cell of HaAP contained one dimer unit corresponding to the biological unit. The monomer structure of HaAP contains a domain comprised of an 11-stranded β-sheet core with 19 surrounding α-helices similar to those of APs from other species, and a unique ‘crown’ domain containing an extended ‘arm’ structure that participates in formation of a hydrophobic cluster at the entrance to the substrate-binding site. The HaAP structure also displays a unique distribution of negatively charged residues and hydrophobic residues in comparison to other known AP structures. AP from Vibrio sp. G15-21 (VAP; a slight halophile) has the highest similarity in sequence (70.0% identity) and structure (C{sup α} r.m.s.d. of 0.82 Å for the monomer) to HaAP. The surface of the HaAP dimer is substantially more acidic than that of the VAP dimer (144 exposed Asp/Glu residues versus 114, respectively), and thus may enable the solubility of HaAP under high-salt conditions. Conversely, the monomer unit of HaAP formed a substantially larger hydrophobic interior

  3. Invited review: The application of alkaline phosphatase assays for the validation of milk product pasteurization.

    Rankin, S A; Christiansen, A; Lee, W; Banavara, D S; Lopez-Hernandez, A


    Standard practices for indirectly assessing the pasteurization status of milk products are primarily based on the thermal inactivation kinetics of the endogenous milk enzyme, alkaline phosphatase (ALP). This assessment provides an invaluable, if not required, tool for both regulatory and in-house process control and validation. Endogenous milk ALP manifests a slightly higher heat resistance than the pathogenic microflora upon which pasteurization time and temperature requirements are based. Hence, ALP activity is recognized and accepted as the method of choice for the rapid validation of milk product pasteurization. However, ALP assays have notable limitations that must be understood if they are to be administered and interpreted correctly and the results are to be applied judiciously. Issues such as the reactivation of heat-denatured ALP and the presence of both heat-stable and -labile microbial ALP are addressed. A discussion of ALP in the milk of nonbovine species is presented based on the limited literature available. Some discussion of research involving alternative pasteurization indicators also is presented. This article is intended to summarize the pertinent details of the ALP assay for dairy products (noting the basis and limitations of various methods) and the processing, handling, and known compositional factors that influence the assay results.

  4. Titanium dioxide nanotube films: Preparation, characterization and electrochemical biosensitivity towards alkaline phosphatase.

    Roman, Ioan; Trusca, Roxana Doina; Soare, Maria-Laura; Fratila, Corneliu; Krasicka-Cydzik, Elzbieta; Stan, Miruna-Silvia; Dinischiotu, Anca


    Titania nanotubes (TNTs) were prepared by anodization on different substrates (titanium, Ti6Al4V and Ti6Al7Nb alloys) in ethylene glycol and glycerol. The influence of the applied potential and processing time on the nanotube diameter and length is analyzed. The as-formed nanotube layers are amorphous but they become crystalline when subjected to subsequent thermal treatment in air at 550°C; TNT layers grown on titanium and Ti6Al4V alloy substrates consist of anatase and rutile, while those grown on Ti6Al7Nb alloy consist only of anatase. The nanotube layers grown on Ti6Al7Nb alloy are less homogeneous, with supplementary islands of smaller diameter nanotubes, spread across the surface. Better adhesion and proliferation of osteoblasts was found for the nanotubes grown on all three substrates by comparison to an unprocessed titanium plate. The sensitivity towards bovine alkaline phosphatase was investigated mainly by electrochemical impedance spectroscopy in relation to the crystallinity, the diameter and the nature of the anodization electrolyte of the TNT/Ti samples. The measuring capacity of the annealed nanotubes of 50nm diameter grown in glycerol was demonstrated and the corresponding calibration curve was built for the concentration range of 0.005-0.1mg/mL.

  5. Alkaline phosphatase levels in diagnostic peritoneal lavage fluid as a predictor of hollow visceral injury.

    Jaffin, J H; Ochsner, M G; Cole, F J; Rozycki, G S; Kass, M; Champion, H R


    Isolated injuries to hollow viscera may result in equivocal diagnostic peritoneal lavage (DPL) findings. Small bowel injuries cause alkaline phosphatase (AP) levels to increase in DPL effluent. The goal of this study was to better define the role of AP levels in the evaluation of the injured abdomen. We prospectively measured AP levels in 672 patients undergoing DPL. These were retrospectively compared with the clinical findings. All 12 patients with small bowel injuries and three of four with large bowel injuries had an AP level > 10 IU/L. There was one patient with an AP level > 10 IU/L without clinically significant intra-abdominal injury. An AP level > 10 IU/L in the DPL effluent predicted injury requiring laparotomy with a specificity of 99.8% and a sensitivity of 94.7%. We recommend using AP levels only in the management of patients with equivocal findings on DPL who would otherwise not undergo laparotomy. This selective use of AP levels will improve the probability of early diagnosis of bowel injury without increasing the cost of care.

  6. Rat enterocytes secrete SLPs containing alkaline phosphatase and cubilin in response to corn oil feeding.

    Mahmood, Akhtar; Shao, Jian-su; Alpers, David H


    Surfactant-like particles (SLP) are unilamellar secreted membranes associated with the process of lipid absorption and isolated previously only from the apical surface of enterocytes. In this paper, the intracellular membrane has been isolated from corn oil-fed animals, identified by its content of the marker protein intestinal alkaline phosphatase (IAP). Another brush-border protein, cubilin, and its anchoring protein megalin have been identified as components of extracellular SLP, but only cubilin is present to any extent in intracellular SLP. During fat absorption, IAP is modestly enriched in intracellular SLP, but full-length cubilin (migrating at 210 kDa in fat-fed mucosal fractions) falls by one-half, although fragments of cubilin are abundant in the intracellular SLP. Both IAP and cubilin colocalize to the same cells during corn oil absorption and colocalize around lipid droplets. This localization is more intense during feeding of corn oil with Pluronic L-81, a detergent that allows uptake of fatty acids and monoglycerides from the lumen, but blocks chylomicron secretion. Confocal microscopy confirms the colocalization of IAP and the ligand for cubilin, intrinsic factor. Possible roles for cubilin in intracellular SLP include facilitating movement of the lipid droplet through the cell and binding to the basolateral membrane before reverse endocytosis.

  7. Differential expression of alkaline phosphatase gene in proliferating primary lymphocytes and malignant lymphoid cell lines.

    Latheef, S A A; Devanabanda, Mallaiah; Sankati, Swetha; Madduri, Ramanadham


    Alkaline Phosphatase (APase) activity has been shown to be enhanced specifically in mitogen stimulated B lymphocytes committed to proliferation, but not in T lymphocytes. APase gene expression was analyzed in proliferating murine and human primary lymphocytes and human malignant cell lines using reverse transcriptase and real time PCR. In mitogen stimulated murine splenic lymphocytes, enhancement of APase activity correlated well with an increase in APase gene expression. However, in mitogen stimulated murine T lymphocytes and human PBL despite a vigorous proliferative response, no increase in APase enzyme activity or gene expression was observed. A constitutive expression of APase activity concomitant with APase gene expression was observed inhuman myeloma cell line, U266 B1. However, neither enzyme activity nor gene expression of APase were observed in human T cell lymphoma, SUPT-1. The results suggest a differential expression of APase activity and its gene in proliferating primary lymphocytes of mice and humans. The specific expression of APase activity and its gene only in human myeloma cells, but not in proliferating primary B cells can be exploited as a sensitive disease marker.

  8. Responses of phosphate transporter gene and alkaline phosphatase in Thalassiosira pseudonana to phosphine.

    Fu, Mei; Song, Xiuxian; Yu, Zhiming; Liu, Yun


    Phosphine, which is released continuously from sediment, can affect the eco-physiological strategies and molecular responses of phytoplankton. To examine the effects of phosphine on phosphorus uptake and utilization in Thalassiosira pseudonana, we examined the transcriptional level of the phosphate transporter gene (TpPHO) and the activity of alkaline phosphatase (AKP) in relation to supplement of various concentrations of phosphine. TpPHO expression was markedly promoted by phosphine in both the phosphate-deficient and phosphate-4 µM culture. However, high phosphine concentrations can inhibit TpPHO transcription in the declining growth phase. AKP activity was also higher in the phosphine treatment groups than that of the control. It increased with increasing phosphine concentration in the range of 0 to 0.056 µM but was inhibited by higher levels of phosphine. These responses revealed that phosphine can affect phosphate uptake and utilization in T. pseudonana. This result was consistent with the effect of phosphine on algal growth, while TpPHO expression and AKP were even more sensitive to phosphine than algal growth. This work provides a basic understanding for further research about how phosphine affects phytoplankton.

  9. Effect of Diazinon on Acid and Alkaline Phosphatase Activities in Plasma and Organs of Clarias gariepinus

    I.R. Inyang


    Full Text Available The aim of this study was to determine the effect of the pesticide, diazinon, on phosphatases in the plasma and organs on Clarias gariepinus. Adult Clarias gariepinus were exposed in four replicates to varying sublethal concentrations diazinon (ranging from 1.00 to 10.0 mg/L in 30-day semi-static bioassays. Alkaline phoshatase (ALP and acid phosphate (ACP were determined in plasma and other organs (gastrointestinal tract - GIT, kidney, muscle, gill and liver of the fish after the experimental exposures. Dizinon did not cause any statistically significant difference on plasma ALP over the concentrations tested (p>0.05, but ACP showed significantly higher mean value at 10 mg/L compared to the control. ALP and ACP values in all the organs (GIT, intestinal tract, kidney, muscle, gill, liver decreased with increasing concentration of diazion. This indicates an evidence of inhibition of these enzymes in the organs by the toxicant, and therefore alteration of biochemical processes in C. gariepinus which can be used as bio-indicators of the effects of diazinon in the Niger Delta environment.

  10. Dynamic evolution of the LPS-detoxifying enzyme intestinal alkaline phosphatase in zebrafish and other vertebrates

    Ye eYang


    Full Text Available Alkaline phosphatases (Alps are well-studied enzymes that remove phosphates from a variety of substrates. Alps function in diverse biological processes, including modulating host-bacterial interactions by dephosphorylating the Gram-negative bacterial cell wall component lipopolysaccharide (LPS. In animals, Alps are encoded by multiple genes characterized by either ubiquitous expression (named Alpls, for their liver expression, or their tissue-specific expression, for example in the intestine (Alpi. We previously characterized a zebrafish alpi gene (renamed here alpi.1 that is regulated by Myd88-dependent innate immune signaling and that is required to prevent a host’s excessive inflammatory reactions to its resident microbiota. Here we report the characterization of two new alp genes in zebrafish, alpi.2 and alp3. To understand their origins, we investigated the phylogenetic history of Alp genes in animals. We find that vertebrate Alp genes are organized in three clades with one of these clades missing from the mammals. We present evidence that these three clades originated during the two vertebrate genome duplications. We show that in zebrafish alpl is ubiquitously expressed, as it is in mammals, whereas the other three alps are specific to the intestine. Our phylogenetic analysis reveals that in contrast to Alpl, which has been stably maintained as a single gene throughout the vertebrates, the Alpis have been lost and duplicated multiple times independently in vertebrate lineages, likely reflecting the rapid and dynamic evolution of vertebrate gut morphologies, driven by changes in bacterial associations and diet.

  11. Enslavement in the water body by toxic Aphanizomenon ovalisporum, inducing alkaline phosphatase in phytoplanktons.

    Bar-Yosef, Yehonathan; Sukenik, Assaf; Hadas, Ora; Viner-Mozzini, Yehudit; Kaplan, Aaron


    The hepatotoxin cylindrospermopsin (CYN) produced by certain cyanobacteria, including Aphanizomenon ovalisporum (hereafter Aphanizomenon) [1], seriously affects lake water quality [2], but its biological role is not known. Strong correlation between Aphanizomenon abundance in Lake Kinneret, Israel, and alkaline phosphatase (APase) activity suggests that inorganic phosphate (Pi) limitation induces the PHO regulon and APase secretion [3]. Staining lake samples with DAPI [4] revealed a high level of polyphosphate bodies (PPB) in Aphanizomenon. Application of enzyme-labeled fluorescence (ELF-APase) [5] showed APase in various organisms, but not in Aphanizomenon. ELF-APase signals and extracellular APase activity in Aphanizomenon were detected only after exploiting PPB under prolonged Pi deprivation in cultures or toward the end of its autumn bloom. Pi deprivation of Aphanizomenon induces CYN production, high-affinity Pi uptake, and an internal, not external, APase. Addition of Aphanizomenon spent media or CYN to various phytoplanktons, including Chlamydomonas reinhardtii, induced genes typically upregulated under Pi limitation and a rise in extracellular APase activity, despite ample surrounding Pi. Coculturing Aphanizomenon with Chlamydomonas or with Debarya sp. showed positive ELF-APase signals, but not in Aphanizomenon. CYN producers promote Pi supply by inducing APase secretion by other phytoplanktons, possibly explaining their increased abundance despite reduced Pi supply from watersheds.

  12. Alkaline Phosphatase: The Next Independent Predictor of the Poor 90-Day Outcome in Alcoholic Hepatitis

    Beata Kasztelan-Szczerbinska


    Full Text Available Aim. Determination of risk factors relevant to 90-day prognosis in AH. Comparison of the conventional prognostic models such as Maddrey’s modified discriminant function (mDF and Child-Pugh-Turcotte (CPT score with newer ones: the Glasgow Alcoholic Hepatitis Score (GAHS; Age, Bilirubin, INR, Creatinine (ABIC score, Model for End-Stage Liver Disease (MELD, and MELD-Na in the death prediction. Patients and Methods. The clinical and laboratory variables obtained at admission were assessed. The mDF, CPT, GAHS, ABIC, MELD, and MELD-Na scores’ different areas under the curve (AUCs and the best threshold values were compared. Logistic regression was used to assess predictors of the 90-day outcome. Results. One hundred sixteen pts fulfilled the inclusion criteria. Twenty (17.4% pts died and one underwent orthotopic liver transplantation (OLT within 90 days of follow-up. No statistically significant differences in the models‘ performances were found. Multivariate logistic regression identified CPT score, alkaline phosphatase (AP level higher than 1.5 times the upper limit of normal (ULN, and corticosteroids (CS nonresponse as independent predictors of mortality. Conclusions. The CPT score, AP > 1.5 ULN, and the CS nonresponse had an independent impact on the 90-day survival in AH. Accuracy of all studied scoring systems was comparable.

  13. Evaluation of biodegradation and biocompatibility of collagen/chitosan/alkaline phosphatase biopolymeric membranes



    The aim of this study was to develop a new variant of membranes based on collagen (COL), chitosan (CHI) and alkaline phosphatase (ALP) immobilized and cross-linking with glutaraldehyde (GA) at different concentrations. The biodegradation in the presence of collagenase was investigated. Biocompatibility was evaluated by MTT assay using a mouse fibroblast cell culture type NCTC (clone 929). Non-cross-linked samples were biocompatible and membranes cross-linked with low concentrations of GA (0.04, 0.08%) were also iocompatible. However, high concentrations of GA lead to a decreased biocompatibility. The adsorption behaviour of Ca$^{2+}$ ions to all membraneswere evaluated using the Freundlich isotherms. Haemolytic studies were performed in order to consider their applications in biomineralization process. By the addition of collagen and ALP to chitosan, the haemolytic indexdecreases, the COL–CHI–ALP membrane being in the non-haemolytic domain, while the COL–CHI–ALP–GA membrane has a haemolytic index greater than 2, and is slightly haemolytic.

  14. Effects of overlying water aeration on phosphorus fractions and alkaline phosphatase activity in surface sediment

    Jianjun Chen; Shaoyong Lu; Yikun Zhao; Wei Wang; Minsheng Huang


    Microbial activity may influence phosphorus (P) deposit and release at the water sediment interface.The properties of DO (dissolved oxygen), pH, P fractions (TP, Ca-P, Fe-P, OP, IP), and APA (alkaline phosphatase activity) at the water sediment interface were measured to investigate microbial activity variations in surface sediment under conditions of two-month intermittent aeration in overlying water.Results showed that DO and TP of overlying water increased rapidly in the first week and then decreased gradually after 15 day of intermittent aeration.Microorganism metabolism in surface sediment increased pH and decreased DO and TP in the overlying water.After two-month intermittent aeration, APA and OP from surface sediment (0-2 crm) were both significantly higher than those from bottom sediment (6-8 cm) (p < 0.05), and surface sediment Fe-P was transferred to OP during the course of microorganism reproduction on the surface sediment.These results suggest that microbial activity and microorganism biomass from the surface sediment were higher than those from bottom sediment afar two-month intermittent aeration in the overlying water.

  15. Substrate and Transition State Binding in Alkaline Phosphatase Analyzed by Computation of Oxygen Isotope Effects.

    Roston, Daniel; Cui, Qiang


    Enzymes are powerful catalysts, and a thorough understanding of the sources of their catalytic power will facilitate many medical and industrial applications. Here we have studied the catalytic mechanism of alkaline phosphatase (AP), which is one of the most catalytically proficient enzymes known. We have used quantum mechanics calculations and hybrid quantum mechanics/molecular mechanics (QM/MM) simulations to model a variety of isotope effects relevant to the reaction of AP. We have calculated equilibrium isotope effects (EIEs), binding isotope effects (BIEs), and kinetic isotope effects (KIEs) for a range of phosphate mono- and diester substrates. The results agree well with experimental values, but the model for the reaction's transition state (TS) differs from the original interpretation of those experiments. Our model indicates that isotope effects on binding make important contributions to measured KIEs on V/K, which complicated interpretation of the measured values. Our results provide a detailed interpretation of the measured isotope effects and make predictions that can test the proposed model. The model indicates that the substrate is deformed in the ground state (GS) of the reaction and partially resembles the TS. The highly preorganized active site preferentially binds conformations that resemble the TS and not the GS, which induces the substrate to adapt to the enzyme, rather than the other way around-as with classic "induced fit" models. The preferential stabilization of the TS over the GS is what lowers the barrier to the chemical step.

  16. Enrichment of thermosensitive chitosan hydrogels with glycerol and alkaline phosphatase for bone tissue engineering applications.

    Douglas, Timothy E L; Krok-Borkowicz, Małgorzata; Macuda, Aleksandra; Pietryga, Krzysztof; Pamuła, Elżbieta


    Thermosensitive injectable chitosan hydrogels can be formed by neutralization of acidic chitosan solutions with sodium betaglycerophosphate (Na-β-GP) coupled with increasing temperature to body temperature. Such hydrogels have been considered for applications in bone regeneration. In this study, chitosan hydrogels were enriched with glycerol and the enzyme alkaline phosphatase (ALP) with a view to improving their suitability as materials for bone tissue engineering. Mineral formation was confirmed by infrared spectroscopy (FTIR) and increases in the mass fraction of the hydrogel not consisting of water. Incorporation of ALP in hydrogels followed by incubation in a solution containing calcium ions and glycerophosphate, a substrate for ALP, led to formation of calcium phosphate within the hydrogel. MG-63 osteoblast-like cells were cultivated in eluates from hydrogels containing ALP and without ALP at different dilutions and directly on the hydrogel samples. Hydrogels containing ALP exhibited superior cytocompatibility to ALP-free hydrogels. These results pave the way for the use of glycerol- and ALP-enriched hydrogels in bone regeneration.

  17. Albumin-to-Alkaline Phosphatase Ratio: A Novel Prognostic Index for Hepatocellular Carcinoma

    Chan, Anthony W. H.; Mo, Frankie K. F.; Wong, Grace L. H.; Wong, Vincent W. S.; Cheung, Yue-Sun; Chan, Henry L. Y.; Yeo, Winnie; Lai, Paul B. S.; To, Ka-Fai


    Prognosis of patients with hepatocellular carcinoma (HCC) depends on both tumour extent and hepatic function reserve. Liver function test (LFT) is a basic routine blood test to evaluate hepatic function. We first analysed LFT components and their associated scores in a training cohort of 217 patients who underwent curative surgery to identify LFT parameters with high performance (discriminatory capacity, homogeneity, and monotonicity of gradient). We derived a novel index, albumin-to-alkaline phosphatase ratio (AAPR), which had the highest c-index (0.646) and χ2 (24.774) among other liver biochemical parameters. The AAPR was an independent prognostic factor for overall and disease-free survival. The adjusted hazard ratio of death and tumour relapse was 2.36 (P = 0.002) and 1.85 (P = 0.010), respectively. The independent prognostic significance of AAPR on top of 5 commonly used and well established staging systems was further confirmed in 2 independent cohorts of patients receiving surgical resection (n = 256) and palliative therapy (n = 425). In summary, the AAPR is a novel index readily derived from a simple low-cost routine blood test and is an independent prognostic indicator for patients with HCC regardless of treatment options. PMID:25737613

  18. Albumin-to-Alkaline Phosphatase Ratio: A Novel Prognostic Index for Hepatocellular Carcinoma

    Anthony W. H. Chan


    Full Text Available Prognosis of patients with hepatocellular carcinoma (HCC depends on both tumour extent and hepatic function reserve. Liver function test (LFT is a basic routine blood test to evaluate hepatic function. We first analysed LFT components and their associated scores in a training cohort of 217 patients who underwent curative surgery to identify LFT parameters with high performance (discriminatory capacity, homogeneity, and monotonicity of gradient. We derived a novel index, albumin-to-alkaline phosphatase ratio (AAPR, which had the highest c-index (0.646 and χ2 (24.774 among other liver biochemical parameters. The AAPR was an independent prognostic factor for overall and disease-free survival. The adjusted hazard ratio of death and tumour relapse was 2.36 (P=0.002 and 1.85 (P=0.010, respectively. The independent prognostic significance of AAPR on top of 5 commonly used and well established staging systems was further confirmed in 2 independent cohorts of patients receiving surgical resection (n=256 and palliative therapy (n=425. In summary, the AAPR is a novel index readily derived from a simple low-cost routine blood test and is an independent prognostic indicator for patients with HCC regardless of treatment options.

  19. Prostatic acid phosphatase in serum and semen of dogs

    CRF Gadelha; WRR Vicente; APC Ribeiro; Apparicio, M. [UNESP; GJ Covizzi; ACN Campos


    The incidence of prostatic malignancy has increased the use of tissue markers to detect cancer. Tissue specific antigens or differentiation antigens are found on the surface of normal cells. Clinically, these antigens are important to diagnose alterations in the tissues and for immunotherapy. The objective of the present study was to evaluate the prostatic acid phosphatase concentration in blood and seminal plasma of intact and healthy dogs at different ages. The evaluation was carried out by...

  20. 中药分期治疗对兔桡骨骨折延迟愈合X线所见及血清ALP的影响%The impact of TCM staging treatment on X-ray and serum alkaline phosphatase of rabbit radius fractures delayed union

    李天朗; 唐志宇; 王祖祥; 付勇; 温月


    目的 观察中药分期治疗对兔骨折延迟愈合X 线所见及血清碱性磷酸酶(ALP)的影响.方法 新西兰兔75 只,左侧无菌术下桡骨中下1/3 切除10 mm 后,除模型组外其余4 组髓腔用骨腊封闭,分为治疗1 组、治疗2 组、治疗3 组、骨折模型组、延迟愈合模型组,每组15 只.其中治疗2 组术后按骨折分期第2 天开始给药,治疗3 组、治疗1 组4 周成模后给药,延迟愈合模型组和骨折模型组术后第2 天灌等容量的生理盐水,延迟愈合模型组成模后骨延迟愈合区注入自体红骨髓.术后第4 周、8 周检测血清ALP 和拍摄X 光片.结果 第4 周治疗2 组的ALP[(155.54±23.33)U/L]明显高于其他4 组;第8 周治疗2 组[(62.44±27.67)U/L]与延迟愈合模型组[(96.72±26.79)U/L]比较差异有统计学意义(P 均<0.01);与骨折模型组比较,差异有统计学意义(P<0.05).与骨折模型组比较,治疗1 组[(74.79± 2.068)U/L]、治疗3 组[(67.27±24.34)U/L]血清ALP 显著下降(P<0.05),治疗2 组无统计学意义(P>0.05).第4 周末,X 线肉眼评分空白对照组高于其他各组;第8 周末治疗2 组肉眼评分显著高于治疗1 组、治疗3 组、延迟愈合模型组(P 均<0.01),但与骨折模型组[(90.56±35.47)U/L]比较,差异无统计学意义(P>0.05).结论 中药分期治疗可提高骨折X 线肉眼评分、治疗2 组血清ALP 水平.%Objective To study the impact of TCM staging treatment on serum alkaline phosphatase of Rabbit radius fractures delayed union. Metheods Left radial bone defect 1 cm were created in 75 New Zealand rabbits and they were divided into 5 group: The first, the second and the third therapeutic group, model group, and delayed union group. The second therapeutic group received herb medicine two days after the operation, the first and the third therapy group received herb medicine four weeks after the operation, the model group and delayed union group received saline solution two days after

  1. Activity of alkaline and acidic phosphatase in glandular cells of uterine endometrium of puerperal ewes after exposure to polychlorinated biphenyls

    Valocky I.; Krajničakova Maria; Legath J.; Lenhardt L.; Ostro A.; Danko J.; Tkačikova L`udmila; Mojžišova Jana; Fialkovičova Maria; Mardzinova Silvia


    The study is focused on the observation of alkaline and acidic phosphatase activity in the glandular cells of uterine endometrium in puerperal ewes after exposure to polychlorinated biphenyls. Ewes of Slovak merino breed (n=25) divided into 2 groups were included in the experiment. The animals in the experimental group (n=14) and control group (n=11) were euthanised on day 17, 25 and 34 postpartum. The ewes in the experimental group were given per os capsules of the chemical preparation Delor...

  2. A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.

    Chen, Wenbo; Liu, Chenxi; Xiao, Yutao; Zhang, Dandan; Zhang, Yongdong; Li, Xianchun; Tabashnik, Bruce E; Wu, Kongming


    Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry) proteins from the bacterium Bacillus thuringiensis (Bt) in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50) of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f) was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects.

  3. A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.

    Wenbo Chen

    Full Text Available Evolution of resistance by insects threatens the continued success of pest control using insecticidal crystal (Cry proteins from the bacterium Bacillus thuringiensis (Bt in sprays and transgenic plants. In this study, laboratory selection with Cry1Ac yielded five strains of cotton bollworm, Helicoverpa armigera, with resistance ratios at the median lethal concentration (LC50 of activated Cry1Ac ranging from 22 to 1700. Reduced activity and reduced transcription of an alkaline phosphatase protein that binds Cry1Ac was associated with resistance to Cry1Ac in the four most resistant strains. A Cry1Ac-binding fragment of alkaline phosphatase from H. armigera (HaALP1f was not toxic by itself, but it increased mortality caused by Cry1Ac in a susceptible strain and in all five resistant strains. Although synergism of Bt toxins against susceptible insects by toxin-binding fragments of cadherin and aminopeptidase N has been reported previously, the results here provide the first evidence of synergism of a Bt toxin by a toxin-binding fragment of alkaline phosphatase. The results here also provide the first evidence of synergism of a Bt toxin by any toxin-binding peptide against resistant insects.

  4. PrognosticValue of PINP,BoneAlkaline Phosphatase, CTX-I, andYKL-40 in Patients With Metastatic Prostate Carcinoma

    Brasso, Klaus; Christensen, Ib Jarle; Johansen, Julia S


    Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma. Prostate. 2006 Apr 1;66(5):503-13. PMID: 16372331 [PubMed - indexed for MEDLINE]......Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma. Prostate. 2006 Apr 1;66(5):503-13. PMID: 16372331 [PubMed - indexed for MEDLINE]...

  5. Mechanistic and Evolutionary Insights from Comparative Enzymology of Phosphomonoesterases and Phosphodiesterases across the Alkaline Phosphatase Superfamily.

    Sunden, Fanny; AlSadhan, Ishraq; Lyubimov, Artem Y; Ressl, Susanne; Wiersma-Koch, Helen; Borland, Jamar; Brown, Clayton L; Johnson, Tory A; Singh, Zorawar; Herschlag, Daniel


    Naively one might have expected an early division between phosphate monoesterases and diesterases of the alkaline phosphatase (AP) superfamily. On the contrary, prior results and our structural and biochemical analyses of phosphate monoesterase PafA, from Chryseobacterium meningosepticum, indicate similarities to a superfamily phosphate diesterase [Xanthomonas citri nucleotide pyrophosphatase/phosphodiesterase (NPP)] and distinct differences from the three metal ion AP superfamily monoesterase, from Escherichia coli AP (EcAP). We carried out a series of experiments to map out and learn from the differences and similarities between these enzymes. First, we asked why there would be independent instances of monoesterases in the AP superfamily? PafA has a much weaker product inhibition and slightly higher activity relative to EcAP, suggesting that different metabolic evolutionary pressures favored distinct active-site architectures. Next, we addressed the preferential phosphate monoester and diester catalysis of PafA and NPP, respectively. We asked whether the >80% sequence differences throughout these scaffolds provide functional specialization for each enzyme's cognate reaction. In contrast to expectations from this model, PafA and NPP mutants with the common subset of active-site groups embedded in each native scaffold had the same monoesterase:diesterase specificities; thus, the >10(7)-fold difference in native specificities appears to arise from distinct interactions at a single phosphoryl substituent. We also uncovered striking mechanistic similarities between the PafA and EcAP monoesterases, including evidence for ground-state destabilization and functional active-site networks that involve different active-site groups but may play analogous catalytic roles. Discovering common network functions may reveal active-site architectural connections that are critical for function, and identifying regions of functional modularity may facilitate the design of new enzymes

  6. Comparative characterization of pulmonary surfactant aggregates and alkaline phosphatase isozymes in human lung carcinoma tissue.

    Iino, Nozomi; Matsunaga, Toshiyuki; Harada, Tsuyoshi; Igarashi, Seiji; Koyama, Iwao; Komoda, Tsugikazu


    Alkaline phosphatase (AP) isozymes are surfactant-associated proteins (SPs). Since several different AP isozymes have been detected in the pneumocytes of lung cancer patients, we attempted to identify the relationship between pulmonary surfactant aggregate subtypes and AP isozymes. Pulmonary surfactant aggregates were isolated from carcinoma and non-carcinoma tissues of patients with non-small cell carcinoma of the lung. Upon analysis, ultraheavy, heavy, and light surfactant aggregates were detected in the non-carcinoma tissues, but no ultraheavy surfactant aggregates were found in the carcinoma tissues. Surfactant-associated protein A (SP-A) was detected as two bands (a 27-kDa band and a 54-kDa band) in the ultraheavy, heavy, and light surfactant aggregates found in the non-carcinoma tissues. Although both SP-A bands were detected in the heavy and light surfactant aggregates from adenocarcinoma tissues, the 54-kDa band was not detected in squamous cell carcinoma tissues. Liver AP (LAP) was detected in the heavy and light surfactant aggregates from both non-carcinoma and squamous carcinoma tissues, but not in heavy surfactant aggregates from adenocarcinoma tissues. A larger amount of bone type AP (BAP) was found in light surfactant aggregate fractions from squamous cell carcinomas than those from adenocarcinoma tissues or non-carcinoma tissues from patients with either type of cancer. LAP, BAP, and SP-A were identified immunohistochemically in type II pneumocytes from non-carcinoma tissues and adenocarcinoma cells, but no distinct SP-A staining was observed in squamous cell carcinoma tissues. The present study has thus revealed several differences in pulmonary surfactant aggregates and AP isozymes between adenocarcinoma tissue and squamous cell carcinoma tissue.

  7. Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation

    Ciancaglini P.


    Full Text Available Endochondral calcification involves the participation of matrix vesicles (MVs, but it remains unclear whether calcification ectopically induced by implants of demineralized bone matrix also proceeds via MVs. Ectopic bone formation was induced by implanting rat demineralized diaphyseal bone matrix into the dorsal subcutaneous tissue of Wistar rats and was examined histologically and biochemically. Budding of MVs from chondrocytes was observed to serve as nucleation sites for mineralization during induced ectopic osteogenesis, presenting a diameter with Gaussian distribution with a median of 306 ± 103 nm. While the role of tissue-nonspecific alkaline phosphatase (TNAP during mineralization involves hydrolysis of inorganic pyrophosphate (PPi, it is unclear how the microenvironment of MV may affect the ability of TNAP to hydrolyze the variety of substrates present at sites of mineralization. We show that the implants contain high levels of TNAP capable of hydrolyzing p-nitrophenylphosphate (pNPP, ATP and PPi. The catalytic properties of glycosyl phosphatidylinositol-anchored, polidocanol-solubilized and phosphatidylinositol-specific phospholipase C-released TNAP were compared using pNPP, ATP and PPi as substrates. While the enzymatic efficiency (k cat/Km remained comparable between polidocanol-solubilized and membrane-bound TNAP for all three substrates, the k cat/Km for the phosphatidylinositol-specific phospholipase C-solubilized enzyme increased approximately 108-, 56-, and 556-fold for pNPP, ATP and PPi, respectively, compared to the membrane-bound enzyme. Our data are consistent with the involvement of MVs during ectopic calcification and also suggest that the location of TNAP on the membrane of MVs may play a role in determining substrate selectivity in this micro-compartment.

  8. Alkaline phosphatase activity related to phosphorus stress of microphytoplankton in different trophic conditions

    Ivančić, Ingrid; Pfannkuchen, Martin; Godrijan, Jelena; Djakovac, Tamara; Marić Pfannkuchen, Daniela; Korlević, Marino; Gašparović, Blaženka; Najdek, Mirjana


    The northern Adriatic (NA) is a favorable basin for studying the adaptive strategies of plankton to a variety of conditions along the steep gradients of environmental parameters over the year. Earlier studies identified phosphorus (P)-limitation as one of the key stresses within the NA that shape the biological response in terms of biodiversity and metabolic adjustments. A wide range of reports supports the notion that P-limitation is a globally important phenomenon in aquatic ecosystems. In this study P stress of marine microphytoplankton was determined at species level along a trophic gradient in the NA. In P-limitation all species with considerable contributions to the diatom community expressed alkaline phosphatase activity (APA), compared to only a few marginal dinoflagellate species. Nevertheless, APA expressing species did not always dominate the phytoplankton community, suggesting that APA is also an important strategy for species to survive and maintain active metabolism outside of their mass abundances. A symbiotic relationship could be supposed for diatoms that did not express APA themselves and probably benefited from APA expressed by attached bacteria. APA was not expressed by any microphytoplankton species during the autumn when P was not limiting, while most of the species did express APA during the P-limitation. This suggests that APA expression is regulated by orthophosphate availability. The methods employed in this study allowed the microscopic detection of APA for each microphytoplankton cell with simultaneous morphologic/taxonomic analysis. This approach uncovered a set of strategies to compete in P-limited conditions within the marine microphytoplankton community. This study confirms the role of P-limitation as a shaping factor in marine ecosystems.

  9. The mechanism of mineralization and the role of alkaline phosphatase in health and disease.

    Orimo, Hideo


    Biomineralization is the process by which hydroxyapatite is deposited in the extracellular matrix. Physiological mineralization occurs in hard tissues, whereas pathological calcification occurs in soft tissues. The first step of mineralization is the formation of hydroxyapatite crystals within matrix vesicles that bud from the surface membrane of hypertrophic chondrocytes, osteoblasts, and odontoblasts. This is followed by propagation of hydroxyapatite into the extracellular matrix and its deposition between collagen fibrils. Extracellular inorganic pyrophosphate, provided by NPP1 and ANKH, inhibits hydroxyapatite formation. Tissue-nonspecific alkaline phosphatase (TNAP) hydrolyzes pyrophosphate and provides inorganic phosphate to promote mineralization. Inorganic pyrophosphate, pyridoxal phosphate, and phosphoethanolamine are thought to be the physiologic substrates of TNAP. These accumulate in the event of TNAP deficiency, e.g., in cases of hypophosphatasia. The gene encoding TNAP is mapped to chromosome 1, consists of 12 exons, and possesses regulatory motifs in the 5'-untranslated region. Inhibition of TNAP enzymatic activity suppresses TNAP mRNA expression and mineralization in vitro. Hypophosphatasia is an inherited systemic bone disease characterized by hypomineralization of hard tissues. The phenotype of hypophosphatasia is varied. To date, more than 200 mutations in the TNAP gene have been reported. Knockout mice mimic the phenotypes of severe hypophosphatasia. Among the mutations in the TNAP gene, c.1559delT is frequent in the Japanese population. This frameshift mutation results in the expression of an abnormally long protein that is degraded in cells. DNA-based prenatal diagnosis using chorionic villus sampling has been developed, but requires thorough genetic counseling. Although hypophosphatasia is untreatable at present, the recent success of enzyme replacement therapy offers promise. The problems presented by impaired mineralization in age

  10. Wnt5a attenuates Wnt3a-induced alkaline phosphatase expression in dental follicle cells

    Sakisaka, Yukihiko [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tsuchiya, Masahiro [Department of Oral Diagnosis, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tohoku Fukushi University, Sendai 989-3201 (Japan); Nakamura, Takashi [Department of Pediatric Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Liason Center for Innovative Dentistry, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Tamura, Masato [Department of Biochemistry and Molecular Biology, Hokkaido University Graduate School of Dentistry, Sapporo 060-8586 (Japan); Shimauchi, Hidetoshi [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan); Nemoto, Eiji, E-mail: [Department of Periodontology and Endodontology, Tohoku University Graduate School of Dentistry, Sendai 980-8575 (Japan)


    Wnt signaling regulates multiple cellular events such as cell proliferation, differentiation, and apoptosis through β-catenin-dependent canonical and β-catenin-independent noncanonical pathways. Canonical Wnt/β-catenin signaling can promote the differentiation of dental follicle cells, putative progenitor cells for cementoblasts, osteoblasts, and periodontal ligament cells, toward a cementoblast/osteoblast phenotype during root formation, but little is known about the biological significance of noncanonical Wnt signaling in this process. We identified the expression of Wnt5a, a representative noncanonical Wnt ligand, in tooth root lining cells (i.e. precementoblasts/cementoblasts) and dental follicle cells during mouse tooth root development, as assessed by immunohistochemistry. Silencing expression of the Wnt5a gene in a dental follicle cell line resulted in enhancement of the Wnt3a (a representative canonical Wnt ligand)-mediated increase in alkaline phosphatase (ALP) expression. Conversely, treatment with recombinant Wnt5a inhibited the increase in ALP expression, suggesting that Wnt5a signaling functions as a negative regulator of canonical Wnt-mediated ALP expression of dental follicle cells. Wnt5a did not affect the nuclear translocation of β-catenin as well as β-catenin-mediated transcriptional activation of T-cell factor (Tcf) triggered by Wnt3a, suggesting that Wnt5a inhibits the downstream part of the β-catenin-Tcf pathway. These findings suggest the existence of a feedback mechanism between canonical and noncanonical Wnt signaling during the differentiation of dental follicle cells. - Highlights: • Dental follicle cells express Wnt5a during tooth root development. • Silencing of Wnt5a enhances Wnt3a-mediated ALP expression of dental follicle cells. • Conversely, treatment with rWnt5a inhibited the increase in ALP expression. • Wnt5a functions as a negative regulator of Wnt3a-mediated ALP expression.

  11. Measurement of alkaline phosphatase in canine seminal plasma--an update.

    Schäfer-Somi, S; Fröhlich, T; Schwendenwein, I


    In dogs, diagnosis of incomplete ejaculation and azoospermia can be made by measuring the activity of the enzyme alkaline phosphatase (AP) in seminal plasma. However, even though upper cut-off value of 5000 IU/l is given in the literature, results by different assays may vary considerably. Furthermore, no data exist concerning the stability of the enzyme during storage of frozen seminal plasma, and no recommendations for pre-analytic dilutions can be found. During the present study, we compared results from a conventional large scale wet chemistry analyzer to a widely used dry chemistry point of care system (POC) and established a best practice for pre-analytical dilutions. Furthermore, stability of enzyme activities in seminal plasma during storage at -18 °C for 24 h was evaluated. The average activity of AP in the 2nd fraction of normal ejaculates measured by Reflotron® was 107,328 IU/l. After 24 h of frozen storage, activities did not differ significantly (96,844 IU / l, p > 0.05). Fresh and frozen samples were analysed in parallel by the POC and conventional chemistry analyser, and the results compared that did not reveal a significant difference (p > 0.05). A dilution of seminal plasma with physiologic saline 1:100 prior to analysis was sufficient for the qualitative information whether AP activity is below or above 5000 IU/l. Present data show that AP measurement by a POC dry chemistry system is sufficiently accurate in diluted seminal plasma for the diagnosis of azoospermia and that seminal plasma can be stored frozen for 24h before analysis.

  12. Using a Personal Glucose Meter and Alkaline Phosphatase for Point-of-Care Quantification of Galactose-1-Phosphate Uridyltransferase in Clinical Galactosemia Diagnosis.

    Zhang, Jingjing; Xiang, Yu; Novak, Donna E; Hoganson, George E; Zhu, Junjie; Lu, Yi


    The personal glucose meter (PGM) was recently shown to be a general meter to detect many targets. Most studies, however, focus on transforming either target binding or enzymatic activity that cleaves an artificial substrate into the production of glucose. More importantly, almost all reports exhibit their methods by using artificial samples, such as buffers or serum samples spiked with the targets. To expand the technology to even broader targets and to validate its potential in authentic, more complex clinical samples, we herein report expansion of the PGM method by using alkaline phosphatase (ALP) that links the enzymatic activity of galactose-1-phosphate uridyltransferase to the production of glucose, which allows point-of-care galactosemia diagnosis in authentic human clinical samples. Given the presence of ALP in numerous enzymatic assays for clinical diagnostics, the methods demonstrated herein advance the field closer to point-of-care detection of a wide range of targets in real clinical samples.

  13. Serum Bone Resorption Markers after Parathyroidectomy for Renal Hyperparathyroidism: Correlation Analyses for the Cross-Linked N-telopeptide of Collagen I and Tartrate-Resistant Acid Phosphatase

    Kuo-Chin Hung


    Full Text Available Patients on long-term dialysis may develop secondary hyperparathyroidism (SHPT with increased serum concentrations of bone resorption markers such as the cross-linked N-telopeptide of type I collagen (NTX and type-5b tartrate-resistant acid phosphatase (TRAP. When SHPT proves refractory to treatment, parathyroidectomy (PTX may be needed. Renal patients on maintenance HD who received PTX for refractory SHPT (n=23 or who did not develop refractory SHPT (control subjects; n=25 were followed prospectively for 4 weeks. Serum intact parathyroid hormone (iPTH, NTX, TRAP, and bone alkaline phosphatase (BAP concentrations were measured serially and correlation analyses were performed. iPTH values decreased rapidly and dramatically. BAP values increased progressively with peak increases observed at 2 weeks after surgery. NTX and TRAP values decreased concurrently and progressively through 4 weeks following PTX. A significant correlation between TRAP and NTX values was observed before PTX but not at 4 weeks after PTX. Additionally, the fractional changes in serum TRAP were larger than those in serum NTX at all times examined after PTX. Serum iPTH, TRAP, and NTX values declined rapidly following PTX for SHPT. Serum TRAP values declined to greater degrees than serum NTX values throughout the 4-week period following PTX.

  14. Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease

    Kriszta Molnár; (A)dám Vannay; Beáta Szebeni; Nóra Fanni Bánki; Erna Sziksz; (A)ron Cseh; Hajnalka Gy(o)rffy


    AIM:To investigate intestinal alkaline phosphatase (iAP) in the intestinal mucosa of children with inflammatory bowel disease (IBD).METHODS:Colonic biopsy samples were taken from 15 newly diagnosed IBD patients and from 10 healthy controls.In IBD patients,specimens were obtained both from inflamed and non-inflamed areas.The iAP mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blotting analysis,respectively.Tissue localization of iAP and Toll-like receptor (TLR) 4 was investigated by immunofluorescent staining.RESULTS:The iAP protein level in the inflamed mucosa of children with Crohn's disease (CD) and ulcerative colitis (UC) was significantly decreased when compared with controls (both P < 0.05).Similarly,we found a significantly decreased level of iAP protein in the inflamed mucosa in CD compared with non-inflamed mucosa in CD (P < 0.05).In addition,the iAP protein level in inflamed colonic mucosa in patients with UC was decreased compared with non-inflamed mucosa in patients with CD (P < 0.05).iAP protein levels in the non-inflamed mucosa of patients with CD were similar to controls.iAP mRNA expression in inflamed colonic mucosa of children with CD and UC was not significantly different from that in non-inflamed colonic mucosa with CD.Expression of iAP mRNA in patients with non-inflamed mucosa and in controls were similar.Co-localization of iAP with TLR4 showed intense staining with a dotted-like pattern.iAP was present in the inflamed and non-inflamed mucosa of patients with CD,UC,and in control biopsy specimens,irrespective of whether it was present in the terminal ileum or in the colon.However,the fluorescent signal of TLR4 was more pronounced in the colon compared with the terminal ileum in all groups studied.CONCLUSION:Lower than normal iAP protein levels in inflamed mucosa of IBD patients may indicate a role for iAP in inflammatory lesions in IBD.Based on our results,administration of exogenous

  15. Alkaline phosphatase activity: new assay for the Reflotron system. Results of the evaluation in eight clinical laboratories.

    Schumann, G; Dominick, H C; Hellmann, D; Klauke, R; Möckesch, M; Stekel, H; von Schenck, H; Kraft, M; Nagel, R; Hänseler, E


    A new reagent carrier, Reflotron ALP, has been developed for the Reflotron system, allowing easy and rapid measurement (in less than 3 minutes) of alkaline phosphatase (ALP) activity in capillary blood, venous blood, heparinized plasma or serum. The evaluation of the analytical performance of the assay was carried out at eight clinical laboratories. The study of the imprecision using the measurements in human samples resulted in coefficients of variation ranging from 1.3% to 4.6% (within-run) and from 3.2% to 4.0% (day-to-day). The analytical specificity of the Reflotron ALP assay agrees well with ALP methods using a N-methyl-D-glucamine buffer solution. The calibration of the Reflotron ALP assay, however, is related to the reference intervals for ALP methods using a diethanolamine buffer solution. Method comparisons were performed with the ALP method on Hitachi instruments using diethanolamine buffer. Reflotron ALP measurements in blood and plasma in 157 randomly selected split samples showed excellent agreement (slope: 0.99; intercept: 0.7 U/l; median bias: 2.3%; median difference from the comparison method: -0.3%). Specimens from pregnant women and adolescents were excluded from this study. Differing values were obtained in a method comparison using 48 samples containing predominantly the ALP bone isoform (slope: 0.81; intercept: 31.5 U/l; median bias: 5.7%; median difference from the comparison method: -12.2%). Regression analysis of the results from 21 sera with prevailing placental ALP gave a slope of 1.51, and an intercept of -41.1 U/l (median bias: 8.6%; median difference from the comparison method: 35.6%). Reflotron ALP was compared with three different wet chemistry procedures using different buffer compounds: N-methyl-D-glucamine or diethanolamine or 2-amino-2-methyl-1-propanol. In samples containing predominantly ALP isoforms not of liver origin, the measurements with N-methyl-D-glucamine buffer gave the best fit with respect to Reflotron. In an

  16. Inhibition of the gut enzyme intestinal alkaline phosphatase may explain how aspartame promotes glucose intolerance and obesity in mice.

    Gul, Sarah S; Hamilton, A Rebecca L; Munoz, Alexander R; Phupitakphol, Tanit; Liu, Wei; Hyoju, Sanjiv K; Economopoulos, Konstantinos P; Morrison, Sara; Hu, Dong; Zhang, Weifeng; Gharedaghi, Mohammad Hadi; Huo, Haizhong; Hamarneh, Sulaiman R; Hodin, Richard A


    Diet soda consumption has not been associated with tangible weight loss. Aspartame (ASP) commonly substitutes sugar and one of its breakdown products is phenylalanine (PHE), a known inhibitor of intestinal alkaline phosphatase (IAP), a gut enzyme shown to prevent metabolic syndrome in mice. We hypothesized that ASP consumption might contribute to the development of metabolic syndrome based on PHE's inhibition of endogenous IAP. The design of the study was such that for the in vitro model, IAP was added to diet and regular soda, and IAP activity was measured. For the acute model, a closed bowel loop was created in mice. ASP or water was instilled into it and IAP activity was measured. For the chronic model, mice were fed chow or high-fat diet (HFD) with/without ASP in the drinking water for 18 weeks. The results were that for the in vitro study, IAP activity was lower (p < 0.05) in solutions containing ASP compared with controls. For the acute model, endogenous IAP activity was reduced by 50% in the ASP group compared with controls (0.2 ± 0.03 vs 0.4 ± 0.24) (p = 0.02). For the chronic model, mice in the HFD + ASP group gained more weight compared with the HFD + water group (48.1 ± 1.6 vs 42.4 ± 3.1, p = 0.0001). Significant difference in glucose intolerance between the HFD ± ASP groups (53 913 ± 4000.58 (mg·min)/dL vs 42 003.75 ± 5331.61 (mg·min)/dL, respectively, p = 0.02). Fasting glucose and serum tumor necrosis factor-alpha levels were significantly higher in the HFD + ASP group (1.23- and 0.87-fold increases, respectively, p = 0.006 and p = 0.01). In conclusion, endogenous IAP's protective effects in regard to the metabolic syndrome may be inhibited by PHE, a metabolite of ASP, perhaps explaining the lack of expected weight loss and metabolic improvements associated with diet drinks.

  17. Versatile and Amplified Biosensing through Enzymatic Cascade Reaction by Coupling Alkaline Phosphatase in Situ Generation of Photoresponsive Nanozyme.

    Jin, Lu-Yi; Dong, Yu-Ming; Wu, Xiu-Ming; Cao, Gen-Xia; Wang, Guang-Li


    The alkaline phosphatase (ALP) biocatalysis followed by the in situ enzymatic generation of a visible light responsive nanozyme is coupled to elucidate a novel amplification strategy by enzymatic cascade reaction for versatile biosensing. The enzymatic hydrolysis of o-phosphonoxyphenol (OPP) to catechol (CA) by ALP is allowed to coordinate on the surface of TiO2 nanoparticles (NPs) due to the specificity and high affinity of enediol ligands to Ti(IV). Upon the stimuli by CA generated from ALP, the inert TiO2 NPs is activated, which demonstrates highly efficient oxidase mimicking activity for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light (λ ≥ 400 nm) irradiation utilizing dissolved oxygen as an electron acceptor. On the basis of the cascade reaction of ALP and the nanozyme of CA coordinated TiO2 (TiO2-CA) NPs, we design exquisitely colorimetric biosensors for probing ALP activity and its inhibitor of 2, 4-dichlorophenoxyacetic acid (2,4-DA). Quantitative probing of ALP activity in a wide linear range from 0.01 to 150 U/L with the detection limit of 0.002 U/L is realized, which endows the methodology with sufficiently high sensitivity for potentially practical applications in real samples of human serum (ALP level of 40-190 U/L for adults). In addition, a novel immunoassay protocol by taking mouse IgG as an example is validated using the ALP/nanozyme cascade amplification reaction as the signal transducer. A low detection limit of 2.0 pg/mL is attained for mouse IgG, which is 4500-fold lower than that of the standard enzyme-linked immuno-sorbent assay (ELISA) kit. Although only mouse IgG is used as a proof-of-concept in our experiment, we believe that this approach is generalizable to be readily extended to other ELISA systems. This methodology opens a new horizon for amplified and versatile biosensing including probing ALP activity and following ALP-based ELISA immunoassays.

  18. Comparison of diazo-coupling, formazan, and silver staining techniques for visualizing alkaline phosphatase isoenzymes after electrophoresis in homogeneous-pore and gradient-pore polyacrylamide gels.

    Hodson, A W; Skillen, A W


    Three techniques for visualization of alkaline phosphatase after polyacrylamide-gel electrophoresis are compared. These are diazo-dye simultaneous coupling with the substrate sodium naphthyl phosphate and 5-chloro-2-toluene diazonium chloride; formazan precipitation with the substrate 5-bromo-4-chloro-3-indolyl phosphate and 3-[4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; and silver staining with the substrate sodium glycerophosphate. Each staining technique was tested with gradient-pore and homogeneous-pore acrylamide-gel electrophoresis. The main factors assessed are sensitivity; separation of the human serum alkaline phosphatase isoenzymes of the liver, bone, and intestinal types; and differences in substrate affinity, as well as the complexity of each technique. Using the three techniques only minor differences in substrate affinity are evident. There is some nonspecific staining with the diazo-coupling technique but not with the formazan and silver staining techniques. The differences, in the mobility of the liver, bone, and intestinal isoenzymes achieved by homogeneous-pore gel electrophoresis are sufficient to allow them to be clearly distinguished. However, only very small differences in mobility are found with gradient-pore gel electrophoresis, but the sharper bands in this medium allow much smaller amounts of activity to be detected. As little as 160 microU of enzyme can be visualized by the diazo technique. Silver staining gives an approximately fourfold increase in sensitivity over the formazan technique, which in turn gives a fourfold increase over the diazo technique. An important aspect of the silver staining technique is the potential of increasing sensitivity much further by improvements in the photographic physical development stage.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.

    Juan Luis Jurat-Fuentes

    Full Text Available Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP were detected by two dimensional differential in-gel electrophoresis (2D-DIGE analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests.

  20. Acute effect of tea, wine, beer, and polyphenols on ecto-alkaline phosphatase activity in human vascular smooth muscle cells.

    Negrão, Maria R; Keating, Elisa; Faria, Ana; Azevedo, Isabel; Martins, Maria J


    Alkaline phosphatase (ALP) is an ecto-enzyme widely distributed across species. It modulates a series of transmembranar transport systems, has an important role in bone mineralization, and can also be involved in vascular calcification. Polyphenol-rich diets seem to have protective effects on human health, namely, in the prevention of cardiovascular diseases. We aimed to investigate the effects of polyphenols and polyphenol-rich beverages upon membranar alkaline phosphatase (ecto-ALP) activity in intact human vascular smooth muscle cells (AALTR). The ecto-ALP activity was determined at pH 7.8, with p-nitrophenyl phosphate as the substrate, by absorbance spectrophotometry at 410 nm. Cell viability was assessed by the lactate dehydrogenase (LDH) method, and the polyphenol content of beverages was assessed using the Folin-Ciocalteu reagent. All polyphenols tested inhibited ecto-ALP activity, in a concentration-dependent way. Teas, wines, and beers also inhibited ecto-ALP activity, largely according to their polyphenol content. All tested compounds and beverages improved or did not change AALTR cell viability. Stout beer was an exception to the described behavior. Although more studies must be done, the inhibition of AALTR ecto-ALP activity by polyphenolic compounds and polyphenol-containing beverages may contribute to their cardiovascular protective effects.

  1. Which one of the two common reporter systems is more suitable for chemiluminescent enzyme immunoassay: alkaline phosphatase or horseradish peroxidase?

    Yu, Songcheng; Yu, Fei; Liu, Lie; Zhang, Hongquan; Zhang, Zhenzhong; Qu, Lingbo; Wu, Yongjun


    Alkaline phosphatase and horseradish peroxidase are the most commonly used reporter systems in chemiluminescent enzyme immunoassay (CLEIA). Which one, therefore, would be better when establishing a CLEIA method for a new target substance? There was no standard answer. In this study, both reporters were compared systematically including luminescence kinetics, conjugation methods, optimal condition and detection performance, using two common drugs, SD-methoxy-pyrimidine and enrofloxacin, as determination objects. The results revealed that there was much difference between the luminescence kinetics of the two systems. However, there was little difference between these systems when detecting the same substance, including in optimal conditions and determination of performance. Both reporters were suitable for establishing chemiluminescent enzyme immunoassays. Therefore, the choice of alkaline phosphatase or horseradish peroxidase as the reporter system in chemiluminescent enzyme immunoassays depends on availability. Conversely, these two report systems could be applied in simultaneous analysis of multicomponents due to their different optical behaviors and similar performances. But attention should be paid to conjugation method and coating buffer, which affected the luminescent intensity of different determination targets.

  2. Serum prostate-specific acid phosphatase: development and validation of a specific radioimmunoassay. [/sup 125/I tracer technique

    Vihko, P.; Sajanti, E.; Jaenne, O.; Peltonen, L.; Vihko, R.


    We describe radioimmunoassay for human prostatic acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC in serum, with use of monospecific antisera raised in rabbits against highly purified acid phosphatase from human prostates. The antiserum did not cross react with partly purified acid phosphatases from human spleen, erythrocytes, or synovial tissues. /sup 125/I-labeled acid phosphatase was prepared by a Chloramine T method, and the bound and free antigen was separated in the assay by use of anti-rabbit gamma-globulin raised in sheep. Uniform low nonspecific binding of the (/sup 125/I)acid phosphatase was achieved by using acid-phosphatase-free serum to prepare standard curves and diluted samples of serum with high acid phosphatase activities. Concentrations of immunoreactive acid phosphatase in the serum of healthy men ranged from <1 to 10 and for 12 patients with advanced prostatic carcinoma between 100 and 500 The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group.

  3. Differentiating Intracellular from Extracellular Alkaline Phosphatase Activity in Soil by Sonication

    Qin, S.P.; Hu, C.S.; Oenema, O.


    Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio

  4. Ultrasensitive detection of cancer cells and glycan expression profiling based on a multivalent recognition and alkaline phosphatase-responsive electrogenerated chemiluminescence biosensor

    Chen, Xiaojiao; He, Yao; Zhang, Youyu; Liu, Meiling; Liu, Yang; Li, Jinghong


    A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode interface, affording fast and highly sensitive ECL cytosensing and cell surface glycan evaluation. Combining the multivalent aptamer interface and ALP nanoprobes, the ECL cytosensor showed a detection limit of 38 CCRF-CEM cells per mL in human serum samples, broad dynamic range and excellent selectivity. In addition, the proposed biosensor provided a valuable insight into dynamic profiling of the expression of different glycans on cell surfaces, based on the carbohydrates recognized by lectins applied to the nanoprobes. This biosensor exhibits great promise in clinical diagnosis and drug screening.A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode

  5. Tissue-nonspecific Alkaline Phosphatase Regulates Purinergic Transmission in the Central Nervous System During Development and Disease

    Álvaro Sebastián-Serrano


    Full Text Available Tissue-nonspecific alkaline phosphatase (TNAP is one of the four isozymes in humans and mice that have the capacity to hydrolyze phosphate groups from a wide spectrum of physiological substrates. Among these, TNAP degrades substrates implicated in neurotransmission. Transgenic mice lacking TNAP activity display the characteristic skeletal and dental phenotype of infantile hypophosphatasia, as well as spontaneous epileptic seizures and die around 10 days after birth. This physiopathology, linked to the expression pattern of TNAP in the central nervous system (CNS during embryonic stages, suggests an important role for TNAP in neuronal development and synaptic function, situating it as a good target to be explored for the treatment of neurological diseases. In this review, we will focus mainly on the role that TNAP plays as an ectonucleotidase in CNS regulating the levels of extracellular ATP and consequently purinergic signaling.

  6. A fluorescence turn on assay for alkaline phosphatase based on the Cu(2+) catalyzed Fenton-like reaction.

    Zhang, Qingfeng; Zhang, Cuiyun; Shahzad, Sohail Anjum; Yu, Cong


    A fluorescence turn-on assay was established for ALP (alkaline phosphatase) based on Cu(2+) catalyzed Fenton-like reaction and Graphene Oxide (GO). GO was utilized to quench the fluorescence of fluorescein (FAM) labeled single strand DNA (F-DNA). ALP can remove the phosphate group in sodium ascorbyl phosphate (SAP), and convert it into reducing ascorbate. Highly reactive hydroxyl radicals (·OH) were generated in the presence of ascorbate and Cu(2+) through the Fenton-like reaction. The reactive radicals generated in situ caused the cleavage of F-DNA into small fragments. When GO was added, the fluorescence emission of the sample without ALP was quenched and fluorescence emission recovered in the presence of ALP. The intensity of the recovered fluorescence was directly related to the concentration of ALP in the assay solution, and a sensitive and selective facile ALP assay is therefore established.

  7. Characterization of alkaline phosphatase activity in seminal plasma and in fresh and frozen-thawed stallion spermatozoa.

    Bucci, Diego; Giaretta, Elisa; Spinaci, Marcella; Rizzato, Giovanni; Isani, Gloria; Mislei, Beatrice; Mari, Gaetano; Tamanini, Carlo; Galeati, Giovanna


    Alkaline phosphatase (AP) has been studied in several situations to elucidate its role in reproductive biology of the male from different mammalian species; at present, its role in horse sperm physiology is not clear. The aim of the present work was to measure AP activity in seminal plasma and sperm extracts from freshly ejaculated as well as in frozen-thawed stallion spermatozoa and to verify whether relationship exists between AP activity and sperm quality parameters. Our data on 40 freshly ejaculated samples from 10 different stallions demonstrate that the main source of AP activity is seminal plasma, whereas sperm extracts contribution is very low. In addition, we found that AP activity at physiological pH (7.0) is significantly lower than that observed at pH 8.0, including the optimal AP pH (pH 10.0). Alkaline phosphatase did not exert any effect on sperm-oocyte interaction assessed by heterologous oocyte binding assay. Additionally, we observed a thermal stability of seminal plasma AP, concluding that it is similar to that of bone isoforms. Positive correlations were found between seminal plasma AP activity and sperm concentration, whereas a negative correlation was present between both spermatozoa extracts and seminal plasma AP activity and seminal plasma protein content. A significant decrease in sperm extract AP activity was found in frozen-thawed samples compared with freshly ejaculated ones (n = 21), concomitantly with the decrease in sperm quality parameters. The positive correlation between seminal plasma AP activity measured at pH 10 and viability of frozen-thawed spermatozoa suggests that seminal plasma AP activity could be used as an additional predictive parameter for stallion sperm freezability. In conclusion, we provide some insights into AP activity in both seminal plasma and sperm extracts and describe a decrease in AP after freezing and thawing.

  8. Active site detection by spatial conformity and electrostatic analysis--unravelling a proteolytic function in shrimp alkaline phosphatase.

    Sandeep Chakraborty

    Full Text Available Computational methods are increasingly gaining importance as an aid in identifying active sites. Mostly these methods tend to have structural information that supplement sequence conservation based analyses. Development of tools that compute electrostatic potentials has further improved our ability to better characterize the active site residues in proteins. We have described a computational methodology for detecting active sites based on structural and electrostatic conformity - CataLytic Active Site Prediction (CLASP. In our pipelined model, physical 3D signature of any particular enzymatic function as defined by its active sites is used to obtain spatially congruent matches. While previous work has revealed that catalytic residues have large pKa deviations from standard values, we show that for a given enzymatic activity, electrostatic potential difference (PD between analogous residue pairs in an active site taken from different proteins of the same family are similar. False positives in spatially congruent matches are further pruned by PD analysis where cognate pairs with large deviations are rejected. We first present the results of active site prediction by CLASP for two enzymatic activities - β-lactamases and serine proteases, two of the most extensively investigated enzymes. The results of CLASP analysis on motifs extracted from Catalytic Site Atlas (CSA are also presented in order to demonstrate its ability to accurately classify any protein, putative or otherwise, with known structure. The source code and database is made available at Subsequently, we probed alkaline phosphatases (AP, one of the well known promiscuous enzymes, for additional activities. Such a search has led us to predict a hitherto unknown function of shrimp alkaline phosphatase (SAP, where the protein acts as a protease. Finally, we present experimental evidence of the prediction by CLASP by showing that SAP indeed has protease activity in

  9. Active site detection by spatial conformity and electrostatic analysis--unravelling a proteolytic function in shrimp alkaline phosphatase.

    Chakraborty, Sandeep; Minda, Renu; Salaye, Lipika; Bhattacharjee, Swapan K; Rao, Basuthkar J


    Computational methods are increasingly gaining importance as an aid in identifying active sites. Mostly these methods tend to have structural information that supplement sequence conservation based analyses. Development of tools that compute electrostatic potentials has further improved our ability to better characterize the active site residues in proteins. We have described a computational methodology for detecting active sites based on structural and electrostatic conformity - CataLytic Active Site Prediction (CLASP). In our pipelined model, physical 3D signature of any particular enzymatic function as defined by its active sites is used to obtain spatially congruent matches. While previous work has revealed that catalytic residues have large pKa deviations from standard values, we show that for a given enzymatic activity, electrostatic potential difference (PD) between analogous residue pairs in an active site taken from different proteins of the same family are similar. False positives in spatially congruent matches are further pruned by PD analysis where cognate pairs with large deviations are rejected. We first present the results of active site prediction by CLASP for two enzymatic activities - β-lactamases and serine proteases, two of the most extensively investigated enzymes. The results of CLASP analysis on motifs extracted from Catalytic Site Atlas (CSA) are also presented in order to demonstrate its ability to accurately classify any protein, putative or otherwise, with known structure. The source code and database is made available at Subsequently, we probed alkaline phosphatases (AP), one of the well known promiscuous enzymes, for additional activities. Such a search has led us to predict a hitherto unknown function of shrimp alkaline phosphatase (SAP), where the protein acts as a protease. Finally, we present experimental evidence of the prediction by CLASP by showing that SAP indeed has protease activity in vitro.

  10. Evaluating the levels of salivary alkaline and acid phosphatase activities as biochemical markers for periodontal disease: A case series

    Sarita Dabra


    Full Text Available Background: The purpose of this study was to determine the salivary levels of alkaline phosphatase (ALP and acid phosphatase (ACP activities in patients with periodontal disease and to evaluate the use of these enzymes as biochemical markers for periodontal tissue damage. Materials and Methods: In this prospective analytical study, we examined the activities of salivary ALP and ACP in patients with periodontal disease, before and after periodontal treatment. The experimental groups consisted of 20 gingivitis patients and 20 periodontitis patients and the control group had healthy subjects (20 samples. The stimulated saliva of the patient was collected in a sterile test tube and analyzed using Hitachi′s Diagnostic Automatic Analyser. Periodontal disease was determined based on clinical parameters such as gingival index, probing depth and clinical attachment loss. Patients with periodontal disease were under conventional periodontal treatment. The statistical analysis applied was Student′s t-test. Probabilities less than 0.05 (P < 0.05 were considered significant. Results: The obtained results showed statistically significant increased activities of ALP and ACP in saliva from patients with periodontal disease in relation to control group. A significant reduction in the enzyme levels was seen after conventional periodontal therapy. Conclusions: Based on these results, salivary ALP and ACP can be considered to be the biomarkers for evaluating periodontal tissue damage.




    The selective targeting of tumours by enzymes conjugated to monoclonal antibodies (mAb) may be an ideal approach to convert relatively nontoxic prodrugs into active agents at the tumour site. We used the anti-carcinoembryonic antigen mAb BW431/26 conjugated to alkaline phosphatase (AP) and phosphory

  12. Labelling of T cell subsets under field conditions in tropical countries. Adaptation of the immuno-alkaline phosphatase staining method for blood smears

    Lisse, I M; Whittle, H; Aaby, P


    Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could...

  13. Evidence of associations between feto-maternal vitamin D status, cord parathyroid hormone and bone-specific alkaline phosphatase, and newborn whole body bone mineral content

    In spite of a high prevalence of vitamin D inadequacy in pregnant women and neonates, relationships among vitamin D status [25(OH)D], parathyroid hormone (PTH), bone specific alkaline phosphatase (BALP), and whole body bone mineral content (WBBMC) in the newborn are poorly characterized. The purpose...

  14. Investigations into the stabilisation of drugs by sugar glasses : II: Delivery of an inulin-stabilised alkaline phosphatase in the intestinal lumen via the oral route

    Eriksson, H.J.C.; Verweij, W.R.; Poelstra, K.; Hinrichs, W.L.J.; de Jong, G.J.; Somsen, G.W.; Frijlink, H.W.


    In this study the possibility to deliver the acid-sensitive enzyme alkaline phosphatase (AP) from calf intestine (CIAP) to the intestinal system by oral administration was investigated. Tablets were prepared and in vitro evaluated. Final proof of concept studies were performed in rats. This acid lab

  15. Changes in morphology of actin filaments and expression of alkaline phosphatase at 3D cultivation of MG-63 osteoblast-like cells on mineralized fibroin scaffolds.

    Goncharenko, A V; Malyuchenko, N V; Moisenovich, A M; Kotlyarova, M S; Arkhipova, A Yu; Kon'kov, A S; Agapov, I I; Molochkov, A V; Moisenovich, M M; Kirpichnikov, M P


    3D cultivation of MG-63 osteoblast-like cells on mineralized fibroin scaffolds leads to an increase in the expression of alkaline phosphatase, an early marker of bone formation. Increased expression is associated with the actin cytoskeleton reorganization under the influence of 3D cultivation and osteogenic calcium phosphate component of the microcarrier.

  16. Characterization of human placental alkaline phosphatase by activity and protein assays, capillary electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

    Eriksson, H J; Somsen, G W; Hinrichs, W L; Frijlink, H W; de Jong, G J


    Placental alkaline phosphatase (PLAP) that had been isolated from human placenta was further purified using subsequent ion-exchange chromatography (IEC), affinity chromatography (AC) and centrifugal membrane concentration (CMC). During the process, the PLAP samples from the different stages of purif

  17. Effect of noise exposure (85 dB ) on testicular adrenocortical steroidogenic key enzymes, acid and alkaline phosphatase activities of sex organs in mature albino rats


    Changes in the activities of △5-3β-hydroysteroid dehydrogenase (HSD) in testis and adrenal gland, 17β-hydroxysteroid dehydrogenase in testis, acid and alkaline phosphatase in testis, prostate and seminal vesicle were observed in noise exposed mature rats at the intensity of 85 dB for 8 h/day for 45 days. The results indicated that noise exposed group showed a significant diminution in the activities of androgenic key enzymes △5-3β and 17β-HSD, acid phosphatase in testis, prostate and seminal vesicle. There was a significant elevation in the activities of adrenal △5-3β-HSD, alkaline phosphatase in testis and other accessory sex organ in noise exposed group. Gonadosomatic, prostatosomatic and seminal vesiculo-somatic indexes were decreased significantly in noise exposed group. Therefore, it is evident that noise exposure at 85dB exerts a deleterious effect on testicular and adrenocortical activities.

  18. The utility of alkaline phosphatase measurement as a screening test for rickets in breast-fed infants and toddlers: a study from the puget sound pediatric research network.

    Taylor, James A; Richter, Monica; Done, Stephen; Feldman, Kenneth W


    To determine if alkaline phosphatase (AP) levels are a useful screening test for rickets, the authors measured serum AP levels in children 6 to 15 months old who were predominantly breast-fed for > 6 months without vitamin D supplementation. Radiographs were obtained on children with elevated AP levels to determine the presence of rickets. AP levels were obtained on 246 children; levels were elevated in 33 (13.4%). Rickets was present in 4 of 18 children with elevated levels on whom radiographs were obtained. The sensitivity and specificity of AP levels as a test for rickets was maximal at a cutoff value of 552 U/L. Using this cutoff value, the specificity of AP levels as a test for rickets was 97.4%, and the positive predictive value (PPV) was 40.0%. These results suggest that AP levels may be a useful screening test for rickets in children who are breast-fed for prolonged periods without vitamin D supplementation.

  19. Assessment of the colorimetric and fluorometric assays for alkaline phosphatase activity in cow's, goat's, and sheep's milk.

    Klotz, V; Hill, Art; Warriner, K; Griffiths, M; Odumeru, J


    Raw milk is a well-established vehicle for the carriage of human pathogens, and many regulatory bodies have consequently mandated compulsory pasteurization as a food safety intervention. The residual activity of alkaline phosphatase (ALP) has historically been used to verify the adequacy of pasteurization of cow's milk. However, there is uncertainty on how the current ALP standards and methods of analysis can be applied to sheep's and goat's milk, which naturally contain different levels of the enzyme than that found in cow's milk. The official ALP methods applied in Canada (colorimetric assay; MFO-3) and in the United States (Fluorophos) were assessed for their ability to detect enzyme activity in raw and pasteurized milk derived from cows, sheep, and goats. The detection limit and the limit of quantitation were 0.8 and 2.02 microg/ml phenol, respectively, for the MFO-3 method and 43 and 85 mU/liter, respectively, for the Fluorophos method. The average ALP levels in raw goat's, cow's, and sheep's milk were 165, 1,562, and 3,512 microg/ml phenol, respectively. Raw milk detection limits, which correspond to raw milk phosphatase levels, were 0.051, 0.485, and 0.023% in cow's, goat's, and sheep's milk, respectively, for the MFO-3 method and 0.007, 0.070, and 0.004%, respectively, for the Fluorophos method. Although both methods can be used for ALP determination in cow's, goat's, and sheep's milk, the Fluorophos assay was superior to the colorimetric MFO-3 method based on sensitivity and time required to complete the analysis.

  20. In vitro induction of alkaline phosphatase levels predicts in vivo bone forming capacity of human bone marrow stromal cells

    Henk-Jan Prins


    Full Text Available One of the applications of bone marrow stromal cells (BMSCs that are produced by ex vivo expansion is for use in in vivo bone tissue engineering. Cultured stromal cells are a mixture of cells at different stages of commitment and expansion capability, leading to a heterogeneous cell population that each time can differ in the potential to form in vivo bone. A parameter that predicts for in vivo bone forming capacity is thus far lacking. We employed single colony-derived BMSC cultures to identify such predictive parameters. Using limiting dilution, we have produced sixteen single CFU-F derived BMSC cultures from human bone marrow and found that only five of these formed bone in vivo. The single colony-derived BMSC strains were tested for proliferation, osteogenic-, adipogenic- and chondrogenic differentiation capacity and the expression of a variety of associated markers. The only robust predictors of in vivo bone forming capacity were the induction of alkaline phosphatase, (ALP mRNA levels and ALP activity during in vitro osteogenic differentiation. The predictive value of in vitro ALP induction was confirmed by analyzing “bulk-cultured” BMSCs from various bone marrow biopsies. Our findings show that in BMSCs, the additional increase in ALP levels over basal levels during in vitro osteogenic differentiation is predictive of in vivo performance.

  1. Changes in Stoichiometry, Cellular RNA, and Alkaline Phosphatase Activity of Chlamydomonas in Response to Temperature and Nutrients

    Hessen, Dag O.; Hafslund, Ola T.; Andersen, Tom; Broch, Catharina; Shala, Nita K.; Wojewodzic, Marcin W.


    Phytoplankton may respond both to elevated temperatures and reduced nutrients by changing their cellular stoichiometry and cell sizes. Since increased temperatures often cause increased thermal stratification and reduced vertical flux of nutrients into the mixed zone, it is difficult to disentangle these drivers in nature. In this study, we used a factorial design with high and low levels of phosphorus (P) and high and low temperature to assess responses in cellular stoichiometry, levels of RNA, and alkaline phosphatase activity (APA) in the chlorophyte Chlamydomonas reinhardtii. Growth rate, C:P, C:N, N:P, RNA, and APA all responded primarily to P treatment, but except for N:P and APA, also temperature contributed significantly. For RNA, the contribution from temperature was particularly strong with higher cellular levels of RNA at low temperatures, suggesting a compensatory allocation to ribosomes to maintain protein synthesis and growth. These experiments suggest that although P-limitation is the major determinant of growth rate and cellular stoichiometry, there are pronounced effects of temperature also via interaction with P. At the ecosystem level, nutrients and temperature will thus interact, but temperatures would likely exert a stronger impact on these phytoplankton traits indirectly via its force on stratification regimes and vertical nutrient fluxes. PMID:28167934

  2. Tissue non-specific alkaline phosphatase production by human dental pulp stromal cells is enhanced by high density cell culture.

    Tomlinson, Matthew J; Dennis, Caitriona; Yang, Xuebin B; Kirkham, Jennifer


    The cell surface hydrolase tissue non-specific alkaline phosphatase (TNAP) (also known as MSCA-1) is used to identify a sub-population of bone marrow stromal cells (BMSCs) with high mineralising potential and is found on subsets of cells within the dental pulp. We aim to determine whether TNAP is co-expressed by human dental pulp stromal cells (hDPSCs) alongside a range of BMSC markers, whether this is an active form of the enzyme and the effects of culture duration and cell density on its expression. Cells from primary dental pulp and culture expanded hDPSCs expressed TNAP. Subsequent analyses revealed persistent TNAP expression and co-expression with BMSC markers such as CD73 and CD90. Flow cytometry and biochemical assays showed that increased culture durations and cell densities enhanced TNAP expression by hDPSCs. Arresting the hDPSC cell cycle also increased TNAP expression. These data confirm that TNAP is co-expressed by hDPSCs together with other BMSC markers and show that cell density affects TNAP expression levels. We conclude that TNAP is a potentially useful marker for hDPSC selection especially for uses in mineralised tissue regenerative therapies.

  3. Effects of synthetic retinoids and retinoic acid isomers on the expression of alkaline phosphatase in F9 teratocarcinoma cells.

    Gianni, M; Zanotta, S; Terao, M; Garattini, S; Garattini, E


    Expression of ALP in F9 teratocarcinoma cells is induced by all-trans retinoic acid (ATRA) (Gianni' et al., Biochem. J. 274: 673-678, 1991). The specific ligand for retinoic acid related receptors (RXRs), 9-cis retinoic acid (9-cis RA), and three synthetic analogs binding to the alpha, beta and gamma forms of the retinoic acid receptors (RARs), AM580, CD2019, and CD437, were used to study their effects on alkaline phosphatase (ALP) enzymatic activity and mRNA levels. At concentrations close to the Kd for their respective receptors, 9-cis RA, AM580 (the RAR alpha agonist) and CD437 (the RAR gamma agonist) clearly upregulate the expression of the ALP gene, whereas the effect of CD2019 (the RAR beta agonist) is very modest. A specific inhibitor of the RAR alpha, Ro 41-5253, completely blocks the induction of ALP triggered by AM580, while it has minor effects on the upregulation caused by ATRA, 9-cis RA, CD437 and CD2019. The induction of ALP observed with the various retinoids is inhibited by the contemporaneous treatment with dibutyryl cAMP. The levels of the RAR alpha and gamma transcripts are unaltered, while RAR beta mRNAs are induced by ATRA, AM580, CD437 and to a lower extent by 9-cis RA and CD2019.

  4. Low dietary copper increases fecal free radical production, fecal water alkaline phosphatase activity and cytotoxicity in healthy men.

    Davis, Cindy D


    One possible dietary factor that may increase susceptibility to colon cancer is inadequate copper intake. The objective of this study was to investigate the effects of low and adequate copper intakes on copper nutriture and putative risk factors for colon cancer susceptibility in healthy men. Seventeen healthy free-living nonsmoking men aged 21-52 y completed a 13-wk controlled feeding study in a randomized crossover design. The basal diet contained 0.59 mg Cu/13.65 MJ. After a 1-wk equilibration period in which the men consumed the basal diet supplemented with 1.0 mg Cu/d, they were randomly assigned to receive either the basal diet or the basal diet supplemented with 2 mg Cu/d for 6 wk. After the first dietary period, the men immediately began to consume the other level of Cu for the last 6 wk. They collected their feces during the equilibration period and during the last 2 wk of the two dietary periods for free radical and fecal water analysis. Low dietary copper significantly (P copper significantly (P copper concentrations but did not affect fecal water volume, pH, iron or zinc concentrations. In contrast to the fecal analysis, hematological indicators of copper status were not significantly affected by the dietary treatments. These results suggest that low dietary copper adversely affects fecal free radical production and fecal water alkaline phosphatase activity, which are putative risk factors for colon cancer.

  5. The relationship between the alkaline phosphatase network and the haematopoiesis in mice subjected to whole-body irradiation

    Almohamad Khaled M.


    Full Text Available Purpose: To investigate the relationship between the alkaline phosphatase (ALP network of the marrow stroma and the haematopoietic regeneration after mice whole-body irradiation. Materials and methods: Three groups of mice were irradiated with a non-lethal ionising radiation dose: the fi rst one received an intraperitoneal injection of Levamisole, ALP inhibitor, 24 h before irradiation; the second one received an intraperitoneal injection of Lisinopril, haematopoiesis inhibitor, 24 h before irradiation; the third was left untreated, but irradiated. The fourth group, untreated and not irradiated, was the control. The total surface occupied by ALP positive processes, revealed by means of ALP cytochemistry in the marrow area, was evaluated semi-quantitively. Nucleated bone marrow cells were also counted. Results: ALP network began to increase 24 h after irradiation to reach a maximum after 72 h, when the bone marrow was almost become completely empty of the haematopoietic cells. This increase advances the haematopoietic recovery. This process was substantially delayed when the mice were injected with Levamisole 24 h before irradiation. On the contrary, ALP network increased strongly since the fi rst day after irradiation when the mice were injected with Lisinopril 24 h before irradiation. Conclusions: These data have indicated that the haematopoietic recovery and repopulation of the bone marrow were advanced by the ALP network recovery.

  6. A High Level of Intestinal Alkaline Phosphatase Is Protective Against Type 2 Diabetes Mellitus Irrespective of Obesity.

    Malo, Madhu S


    Mice deficient in intestinal alkaline phosphatase (IAP) develop type 2 diabetes mellitus (T2DM). We hypothesized that a high level of IAP might be protective against T2DM in humans. We determined IAP levels in the stools of 202 diabetic patients and 445 healthy non-diabetic control people. We found that compared to controls, T2DM patients have approx. 50% less IAP (mean +/- SEM: 67.4 +/- 3.2 vs 35.3 +/- 2.5 U/g stool, respectively; p diabetes status. With each 25 U/g decrease in stool IAP, there is a 35% increased risk of diabetes. The study revealed that obese people with high IAP (approx. 65 U/g stool) do not develop T2DM. Approx. 65% of the healthy population have diabetes', and might develop T2DM and other metabolic disorders in the near future. In conclusion, high IAP levels appear to be protective against diabetes irrespective of obesity, and a 'temporal IAP profile' might be a valuable tool for predicting 'the incipient metabolic syndrome', including 'incipient diabetes'.

  7. Ultra-sensitive conductometric detection of heavy metals based on inhibition of alkaline phosphatase activity from Arthrospira platensis.

    Tekaya, Nadèje; Saiapina, Olga; Ben Ouada, Hatem; Lagarde, Florence; Ben Ouada, Hafedh; Jaffrezic-Renault, Nicole


    This study is based on the conductometric measurement of alkaline phosphatase activity (APA) from the cyanobacterium, Arthrospira platensis, called Spirulina. Cyanobacterium cells were directly immobilized, by physical adsorption, on the ceramic part of gold interdigitated transducers. This activity was inhibited in the presence of heavy metals and a variation of the local conductivity was measured after addition of the substrate. The Michaelis-Menten constant (Km) was evaluated to be 0.75 mM through a calibration curve of the substrate, disodium 4-nitrophenylphosphate p-nitrophenyl phosphate (pNPP). Inhibition of APA was observed with cadmium and mercury with a detection limit of 10(-20) M. The half maximal inhibitory concentration (IC50) was determined at 10(-19) M for Cd(2+) and 10(-17) M for Hg(2+), and the binding affinity of heavy metal (Ki) was equal to the IC50. On the sensor surface, scanning electron microscopy (SEM) images revealed a remarkable evolution of the cyanobacterium's external surface that was attributable to the first defense mechanism against toxic heavy metals in trace. This effect was also confirmed through the important increase of response time τ(90%) recorded for APA response towards the substrate pNPP after cell exposure to metallic cations. Lifetime of the Spirulina-based biosensor was estimated to be more than 25 days.

  8. Effects of garlic and diallyl trisulfide on the growth, photosynthesis, and alkaline phosphatase activity of the toxic cyanobacterium Microcystis aeruginosa.

    Wang, Shoubing; Wang, Yuanan; Ma, Xiaoxue; Xu, Ziran


    To identify a botanical algicide and elucidate the response of cyanobacteria to the extract from higher plants, the effects of garlic and garlic-derived diallyl trisulfide on Microcystis aeruginosa were studied. Effects were evaluated by changes in cell density, chlorophyll a, maximum effective quantum yield (Fv/Fm), effective quantum yield (YII), non-photochemical quenching (NPQ), and rapid light curves of M. aeruginosa. In addition, alkaline phosphatase activity (APA) was measured when M. aeruginosa was incubated with diallyl trisulfide. Results indicated that the inhibition by garlic and diallyl trisulfide was significant. The 120-h 50 % effective concentrations of garlic and diallyl trisulfide (EC50) were 0.75 g L(-1) and 2.84 mg L(-1), respectively. Moreover, the inhibitory rate increased with increasing concentration and the growth of M. aeruginosa was inhibited by 90.0 % at the highest concentrations. We also show that the response of M. aeruginosa to stress could involve both impairment of the photosynthetic center PSII and alteration of APA. For example, at high garlic concentration (2.0 g L(-1)), Fv/Fm significantly decreased from 0.501 to 0.084 (p garlic as an environmentally friendly algicide.

  9. Dietary free fatty acids form alkaline phosphatase-enriched microdomains in the intestinal brush border membrane

    Hansen, Gert H; Rasmussen, Karina; Niels-Christiansen, Lise-Lotte;


    -linked enzyme is the membrane protein in the brush border with the highest affinity for lipid rafts, this implies that free fatty acids selectively insert stably into these membrane microdomains. We have previously shown that absorption of dietary lipids transiently induce a selective endocytosis of AP from...... the brush border, and from work by others it is known that fat absorption is accompanied by a rise in serum AP and secretion of surfactant-like particles from enterocytes. We propose that these physiological processes may be triggered by the sequestering of dietary free fatty acids in lipid raft......Free fatty acids released during intralumenal digestion of dietary fat must pass through the enterocyte brush border membrane before triacylglycerol reassembly and subsequent chylomicron delivery to the lymph system. In the present work fluorescent BODIPY fatty acid analogs were used to study...

  10. Evaluation of alkaline phosphatase activity and availability of various P fractions for bean (Phaseolus vulgaris in some calcareous soils amended with municipal sewage sludge

    T. Raeisi


    Full Text Available To evaluate the relationship of various P fractions and alkaline phosphatase activity with bean indices growing in 10 calcareous soils, amended with municipal sewage sludge from Chaharmahal-Va-Bakhtiari province, a greenhouse research was carried out. Soil samples were incubated for one month with sludge at a rate equivalent to 1% (w/w. Then, the P fractions, including P adsorbed by Fe and Al oxides (]NaOH+CB]-P, occluded P (CBD-P and P absorbed by Ca (HCl-P, were determined by Olsen and Summers' sequential fractionation procedure. Furthermore, total P, organic P and residual P were determined. Also, alkaline phosphatase activity was measured. A pot experiment in a completely randomized design with three replications in the ten soils was done to evaluate the bean plant indices. The results showed that the amount of P fractions decreased in the following order: HCl-P>residual-P>]NaOH+CB]-P > OP>CBD-P. The results also indicated that alkaline phosphatase activity was significantly correlated with CBD-P fraction, organic P and total P. In addition, significant correlations were found between ([NaOH+CB]-P and HCl-P and plant shoots. In general, the results of this research showed that P fractionation method appears to be a powerful tool to identify the P status and availability in the soils amended with sewage sludge.

  11. Acid and alkaline phosphatase activities in a novel phosphorothionate (RPR-11) treated male and female rats. Evidence of dose and time-dependent response.

    Rahman, M F; Siddiqui, M K; Jamil, K


    The effect of a novel phosphorothionate, the methyl ester of 2-butenoic acid-3-diethoxy phosphinothioyl (RPR-II) was studied on membrane bound target enzymes Acid (AcP) and Alkaline (AkP) Phosphatases in different tissues of male and female albino Wistar rats. Three sub-chronic doses 0.014 (low), 0.028 (medium) and 0.042 (high)mg/kg-1 were administered to the rats daily for a period of 90 days. The long term and repeated administration of RPR-II caused significant increase of AcP and AkP in serum and kidney (AcP), whereas these enzymes simultaneously decreased significantly in liver, kidney (female rat AkP) and lung tissues in both male and female rats after 45 and 90 days of treatment. However, the kidney AcP increased significantly in both the sexes which is suggestive of an increase in synthesis of this enzyme which may be an adaptive mechanism to the toxicant stress. The changes in serum, liver, kidney and lung of both male and female rats by this compound were statistically significant when compared with two way Anova showing that they are dose and time dependent. The alterations in male rats were statistically insignificant when compared with female rats showing no sexual dimorphism by this compound. Recovery was observed after 28 days of post treatment (withdrawal study) indicating reversal of the toxic symptoms once the toxicant is removed. High degree negative correlation was observed for serum versus liver and lung and in other cases substantial correlation was observed. The changes observed in these enzymes showed that liver was most susceptible followed by lung and kidney. There are marker enzymes and their increase in different tissues might be due to the increased permeability of plasma membrane or cellular necrosis, showing the stress condition of the treated rats. This investigation elucidates the effect of these biomarker enzymes which increased in blood, might be due to the necrosis of liver, kidney and lung tissues by this compound.

  12. Effects of Dihydroartemisinin and Artemether on the Growth, Chlorophyll Fluorescence, and Extracellular Alkaline Phosphatase Activity of the Cyanobacterium Microcystis aeruginosa

    Wang, Shoubing; Xu, Ziran


    Increased eutrophication in the recent years has resulted in considerable research focus on identification of methods for preventing cyanobacterial blooms that are rapid and efficient. The objectives of this study were to investigate the effects of dihydroartemisinin and artemether on the growth of Microcystis aeruginosa and to elucidate its mode of action. Variations in cell density, chlorophyll a, soluble protein, malondialdehyde, extracellular alkaline phosphatase activity (APA), and chlorophyll fluorescence parameters (Fv/Fm, ΦPSII, ETR, rapid light curves, fast chlorophyll fluorescence curves on fluorescence intensity, and relative variable fluorescence) were evaluated by lab-cultured experiments. Our results demonstrated that both dihydroartemisinin and artemether inhibited the growth of M.aeruginosa by impairing the photosynthetic center in photosystem II and reducing extracellular APA, with a higher sensitivity exhibited toward artemether. The inhibitory effects of dihydroartemisinin on M.aeruginosa increased with concentration, and the maximum growth inhibitory rate was 42.17% at 24 mg·L-1 after 120h exposure, whereas it was 55.72% at 6 mg·L-1 artemetherafter 120h exposure. Moreover, the chlorophyll fluorescence was significantly inhibited (p<0.05) after 120h exposure to 12 and 24 mg·L-1 dihydroartemisinin. Furthermore, after 120h exposure to 6 mg·L-1 artemether, Fv/Fm, ΦPSII, ETR and rETRmax showed a significant decrease (p<0.01) from initial values of 0.490, 0.516, 17.333, and 104.800, respectively, to 0. One-way analysis of variance showed that 6 mg·L-1 artemether and 24 mg·L-1 dihydroartemisinin had significant inhibitory effects on extracellular APA (p<0.01). The results of this study would be useful to further studies to validate the feasibility of dihydroartemisinin and artemether treatment to inhibit overall cyanobacterial growth in water bodies, before this can be put into practice. PMID:27755566

  13. The 1.4 A crystal structure of the large and cold-active Vibrio sp. alkaline phosphatase.

    Helland, Ronny; Larsen, Renate Lie; Asgeirsson, Bjarni


    Alkaline phosphatase (AP) from the cold-adapted Vibrio strain G15-21 is among the AP variants with the highest known k(cat) value. Here the structure of the enzyme at 1.4 A resolution is reported and compared to APs from E. coli, human placenta, shrimp and the Antarctic bacterium strain TAB5. The Vibrio AP is a dimer although its monomers are without the long N-terminal helix that embraces the other subunit in many other APs. The long insertion loop, previously noted as a special feature of the Vibrio AP, serves a similar function. The surface does not have the high negative charge density as observed in shrimp AP, but a positively charged patch is observed around the active site that may be favourable for substrate binding. The dimer interface has a similar number of non-covalent interactions as other APs and the "crown"-domain is the largest observed in known APs. Part of it slopes over the catalytic site suggesting that the substrates may be small molecules. The catalytic serines are refined with multiple conformations in both monomers. One of the ligands to the catalytic zinc ion in binding site M1 is directly connected to the crown-domain and is closest to the dimer interface. Subtle movements in metal ligands may help in the release of the product and/or facilitate prior dephosphorylation of the covalent intermediate. Intersubunit interactions may be a major factor for promoting active site geometries that lead to the high catalytic activity of Vibrio AP at low temperatures.

  14. Unraveling the Alkaline Phosphatase Inhibition, Anticancer, and Antileishmanial Potential of Coumarin-Triazolothiadiazine Hybrids: Design, Synthesis, and Molecular Docking Analysis.

    Ibrar, Aliya; Zaib, Sumera; Jabeen, Farukh; Iqbal, Jamshed; Saeed, Aamer


    A series of new coumarin-triazolothiadiazine hybrid compounds (5a-j) was designed and synthesized by using the molecular hybridization concept. The cyclocondensation reaction involves the coumarinyl 4-amino-1,2,4-triazole and a range of bromo-acetophenones, delivering the desired products in good yields. The structures of the synthesized compounds were established on the basis of spectro-analytical data. The prepared compounds were evaluated against alkaline phosphatase (ALP) where compound 5j incorporating bis-coumarinyl motifs at the 3- and 6-positions of the heteroaromatic core turned out to be a potent inhibitor with an IC50 value of 1.15 ± 1.0 µM. The synthesized compounds were also tested against Leishmania major and 5h was the lead member with an IC50 value of 0.89 ± 0.08 μM. Anticancer activity was also determined using kidney fibroblast (BHK-21) and lung carcinoma (H-157) cancer cell lines. Compound 5i showed highest cytotoxic potential against H-157 cells with an IC50 value of 1.01 ± 0.12 μM, which is an improved inhibition compared to the standards (vincristine and cisplatin) used in this assay. Molecular docking studies were carried out on the synthesized library of coumarin-triazolothiadiazine hybrids against ALP. Almost all of the compounds showed strong interactions with the key residues of the active site of the receptor. In case of compounds 5a-c, 5h, and 5j, docking results positively complemented the experimental screening. These results provided substantial evidence for the further development of these compounds as potent inhibitors of ALP.

  15. Occurrence of matrix-bound phosphine in polar ornithogenic tundra ecosystems: effects of alkaline phosphatase activity and environmental variables.

    Zhu, Renbin; Ma, Dawei; Ding, Wei; Bai, Bo; Liu, Yashu; Sun, Jianjun


    Phosphine (PH(3)), a reduced phosphorus compound, is a highly toxic and reactive atmospheric trace gas. In this study, a total of ten ornithogenic soil/sediment profiles were collected from tundra ecosystems of east Antarctica and Arctic, and matrix-bound phosphine (MBP), the phosphorus fractions and alkaline phosphatase activity (APA) were analyzed. High MBP concentrations were found in these profiles with the range from 39.59 ng kg(-1) dw to 11.77 μg kg(-1) dw. MBP showed a consistent vertical distribution pattern in almost all the soil profiles, and its concentrations increased at soil surface layers and then decreased with depths. MBP levels in the ornithogenic soils were two to three orders of magnitude lower than those in ornithogenic sediments. The yield of PH(3) as a fraction of total P in all the profiles ranged from 10(-5) to 10(-9) mgPH(3) mg(-1)P with higher mean PH(3) yield in the ornithogenic sediments. The ornithogenic soils showed high concentrations of total phosphorus (TP), organic phosphorus (OP), inorganic phosphorus (IP) and metal elements (Cu, Zn, Mn, Fe, Al and Ca) but low MBP levels, vice versa for the ornithogenic sediments. No correlation had been obtained between MBP concentrations and IP, OP and TP. There existed an exponential correlation (r=0.67, psoil/sediment moisture. MBP concentrations showed a significant positive correlation with APA (r=0.668, psoils/sediments. Our results indicated that MBP is an important gaseous link in the phosphorus biogeochemical cycles of ornithogenic tundra ecosystems in Antarctica and Arctic.

  16. One-step immunoassay for tetrabromobisphenol a using a camelid single domain antibody-alkaline phosphatase fusion protein.

    Wang, Jia; Majkova, Zuzana; Bever, Candace R S; Yang, Jun; Gee, Shirley J; Li, Ji; Xu, Ting; Hammock, Bruce D


    Tetrabromobisphenol A (TBBPA) is a ubiquitous brominated flame retardant, showing widespread environmental and human exposures. A variable domain of the heavy chain antibody (VHH), naturally occurring in camelids, approaches the lower size limit of functional antigen-binding entities. The ease of genetic manipulation makes such VHHs a superior choice to use as an immunoreagent. In this study, a highly selective anti-TBBPA VHH T3-15 fused with alkaline phosphatase (AP) from E. coli was expressed, showing both an integrated TBBPA-binding capacity and enzymatic activity. A one-step immunoassay based on the fusion protein T3-15-AP was developed for TBBPA in 5% dimethyl sulfoxide (DMSO)/phosphate buffered saline (PBS, pH 7.4), with a half-maximum signal inhibition concentration (IC50) of 0.20 ng mL(-1). Compared to the parental VHH T3-15, T3-15-AP was able to bind to a wider variety of coating antigens and the assay sensitivity was slightly improved. Cross-reactivity of T3-15-AP with a set of important brominated analogues was negligible (<0.1%). Although T3-15-AP was susceptible to extreme heat (90 °C), much higher binding stability at ambient temperature was observed in the T3-15-AP-based assay for at least 70 days. A simple pretreatment method of diluting urine samples with DMSO was developed for a one-step assay. The recoveries of TBBPA from urine samples via this one-step assay ranged from 96.7% to 109.9% and correlated well with a high-performance liquid chromatography-tandem mass spectroscopy (HPLC-MS/MS) method. It is expected that the dimerized fusion protein, VHH-AP, will show promising applications in human exposure and environmental monitoring.

  17. A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae.

    Fernandez, Luisa E; Aimanova, Karlygash G; Gill, Sarjeet S; Bravo, Alejandra; Soberón, Mario


    A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide-displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI-ALP (GPI-anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI-ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI-ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI-ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae.

  18. Ejaculation training, seminal alkaline phosphatase and semen preservation through cooling in a milk-based extender in domestic cats.

    Valiente, Carla; de la Sota, Pablo E; Arauz, Sandra; Gobello, Cristina


    The purpose of this report is to describe (1) the training of domestic cats in ejaculation into an artificial vagina (AV), (2) alkaline phosphatase (AP) concentrations in whole ejaculates, and (3) the in vitro effect of a skimmed-milk plus egg yolk (SM-Y) extender on feline spermatozoa incubated at 4ºC. Five post-pubertal cats were trained to ejaculate into an AV three times a week for 20 mins in the presence of a teaser queen. Fifty AV-obtained ejaculates were macro- and microscopically assessed, and the AP therein measured by optimized colorimetry. Eighty AV-obtained ejaculates were pooled, diluted in SM-Y extender [80% (v/v) skimmed milk, 20% (v/v) egg yolk, and antibiotics], stored at 4°C and evaluated daily for 6 days. All the animals could be trained to ejaculate, although the interval up to the first AV ejaculation varied from 1.5 to 5.5 months (mean 3.9 months). The final performance at collection ranged from excellent to poor and was inversely related to the training period required in all cases. The mean AP concentration in whole ejaculates was 20,645.6 ± 4405U/l, which was not correlated with the concentration of spermatozoa. Most seminal parameters [(%); total (77 ± 2.3) and progressive (62.7 ± 3.4) motility, live sperm (91.8 ± 1.2), intact plasmalemma (83.5 ± 2.6), normal acrosomes (83.5 ± 2.6), pH (6.6 ± 0.0) and osmolarity (mOsm/l; 321 ± 5.2)], though decreasing during storage in the cold, remained within values compatible with in vivo fertilization for 2 days.

  19. ALP (Alkaline Phosphatase) Test

    ... Paget's disease or other bone conditions, such as vitamin D deficiency. If ALP results are increased but ... be seen temporarily after blood transfusions or heart bypass surgery. A deficiency in zinc may cause decreased ...

  20. Presence and patterns of alkaline phosphatase activity and phosphorus cycling in natural riparian zones under changing nutrient conditions

    Peifang Wang


    Full Text Available Phosphorus (P is an important limiting nutrient in aquatic ecosystems and knowledge of P cycling is fundamental for reducing harmful algae blooms and other negative effects in water. Despite their importance, the characteristics of P cycling under changing nutrient conditions in shallow lakes were poorly investigated. In this study, in situ incubation experiments were conducted in a natural riparian zone in the main diversion channel used for water transfer into Lake Taihu (Wangyu River. Variations in microbial biomass, dissolved P fractions (organic and inorganic, and alkaline phosphatase activity (bulk APA and specific APA were determined after incubation with and without the addition of P and nitrogen (N (4 total water treatments: +P, +N, +NP, and control. Experiments were conducted during two seasons (late spring and early fall to account for natural differences in nutrient levels that may occur in situ. Our results demonstrated that low levels of DRP may not necessarily indicate P limitation. Phytoplankton exhibited “serial N limitation with P stress” in May, such that chlorophyll a (Chl a increased significantly with N addition, while the limiting nutrient shifted to P in October and phytoplankton biomass increased with P addition. Phytoplankton contributed greatly to APA production and was significantly influenced by P bioavailability, yet high levels of bulk APA were also not necessarily indicative of P limitation. In contrast to phytoplankton, bacteria were less P stressed. As a consequence of enhanced utilization of dissolved reactive P (DRP and dissolved organic P (DOP, +N treatment elevated APA significantly. By contrast, APA could be repressed to low values and phytoplankton converted a large portion of DRP to DOP with P addition. But this was not consistent with bacteria APA (bact-APA in the absence or presence of abundant phytoplankton biomass. The correlation between bulk APA and DRP was good at separate sites and discrepant

  1. Enzymatically mediated bioprecipitation of heavy metals from industrial wastes and single ion solutions by mammalian alkaline phosphatase.

    Chaudhuri, Gouri; Shah, Gaurav A; Dey, Pritam; S, Ganesh; Venu-Babu, P; Thilagaraj, W Richard


    The study was aimed at investigating the potential use of calf intestinal alkaline phosphatase (CIAP) enzyme in the removal of heavy metals (Cd(2+), Ni(2+), Co(2+) and Cr(3+/6+)) from single ion solutions as well as tannery and electroplating effluents. CIAP mediated bioremediation (white biotechnology) is a novel technique that is eco-friendly and cost effective unlike the conventional chemical technologies. Typical reactions containing the enzyme (CIAP) and p-nitrophenyl phosphate (pNPP) as substrate in Tris-HCl buffer (pH 8 and 11) and either single ion metal solutions (250 ppm and 1000 ppm) or effluents from tannery or electroplating industry were incubated at 37°C for 30 min, 60 min and 120 min. The inorganic phosphate (P(i)) generated due to catalytic breakdown of pNPP complexes free metal ions as metal-phosphate and the amount of metal precipitated was derived by estimating the reduction in the free metal ion present in the supernatant of reactions employing atomic absorption spectrophotometer (AAS). Better precipitation of metal was obtained at pH 11 than at pH 8 and between the two concentrations of different metals tested, an initial metal concentration of 250 ppm in the reaction gave more precipitation than with 1000 ppm. Experimental data showed that at pH 11, the percentage of removal of metal ions (for an initial concentration of 250 ppm) was in the following order: Cd(2+) (80.99%) > Ni(2+) (64.78%) > Cr(3+) > (46.15%) > Co(2+) (36.47%) > Cr(6+) (32.33%). The overall removal of Cr(3+) and Cr(6+) from tannery effluent was 32.77% and 37.39% respectively in 120 min at pH 11. Likewise, the overall removal of Cd(2+), Co(2+) and Ni(2+) from electroplating effluent was 50.42%, 13.93% and 38.64% respectively in 120 min at pH 11. The study demonstrates that bioprecipitation by CIAP may be a viable and environmental friendly method for clean-up of heavy metals from tannery and electroplating effluents.

  2. The cellular prion protein interacts with the tissue non-specific alkaline phosphatase in membrane microdomains of bioaminergic neuronal cells.

    Myriam Ermonval

    Full Text Available BACKGROUND: The cellular prion protein, PrP(C, is GPI anchored and abundant in lipid rafts. The absolute requirement of PrP(C in neurodegeneration associated to prion diseases is well established. However, the function of this ubiquitous protein is still puzzling. Our previous work using the 1C11 neuronal model, provided evidence that PrP(C acts as a cell surface receptor. Besides a ubiquitous signaling function of PrP(C, we have described a neuronal specificity pointing to a role of PrP(C in neuronal homeostasis. 1C11 cells, upon appropriate induction, engage into neuronal differentiation programs, giving rise either to serotonergic (1C11(5-HT or noradrenergic (1C11(NE derivatives. METHODOLOGY/PRINCIPAL FINDINGS: The neuronal specificity of PrP(C signaling prompted us to search for PrP(C partners in 1C11-derived bioaminergic neuronal cells. We show here by immunoprecipitation an association of PrP(C with an 80 kDa protein identified by mass spectrometry as the tissue non-specific alkaline phosphatase (TNAP. This interaction occurs in lipid rafts and is restricted to 1C11-derived neuronal progenies. Our data indicate that TNAP is implemented during the differentiation programs of 1C11(5-HT and 1C11(NE cells and is active at their cell surface. Noteworthy, TNAP may contribute to the regulation of serotonin or catecholamine synthesis in 1C11(5-HT and 1C11(NE bioaminergic cells by controlling pyridoxal phosphate levels. Finally, TNAP activity is shown to modulate the phosphorylation status of laminin and thereby its interaction with PrP. CONCLUSION/SIGNIFICANCE: The identification of a novel PrP(C partner in lipid rafts of neuronal cells favors the idea of a role of PrP in multiple functions. Because PrP(C and laminin functionally interact to support neuronal differentiation and memory consolidation, our findings introduce TNAP as a functional protagonist in the PrP(C-laminin interplay. The partnership between TNAP and PrP(C in neuronal cells may

  3. Osteoblast response (initial adhesion and alkaline phosphatase activity following exposure to a barrier membrane/enamel matrix derivative combination

    Thangakumaran S


    Full Text Available Background and Objective: The enamel matrix derivative (EMD has been used in combination with barrier membranes to optimize regeneration in vertical osseous defects. However, the osteoblast response when exposed to the EMD/barrier membrane combination has not yet been evaluated. The osteoblast behavior when exposed to a combination of regenerative materials must be evaluated to fully understand their effect on bone regeneration. Therefore, the present study was undertaken to estimate the initial adhesion and alkaline phosphatase (ALP activity of an osteoblast cell line (SaOS-2 when exposed to four commercially available resorbable membranes and determine if the addition of EMD had any modulatory effect on osteoblast behavior. Materials and Methods: 5 x 104 SaOS-2 cells between passages 7-10 were cultured in two 24-well culture plates. Plate A was used for the adhesion assay and Plate B was used for the ALP assay. A MTT (3-[4, 5-dimethylthiazolyl-2]-2, 5-diphenyltetrazolium bromide assay was done after 24 hours to determine the adhesion of the osteoblastic cells to four barrier membranes: 1 a non cross-linked porcine Type I and III collagen membrane (BG, 2 a weakly cross-linked Type I collagen membrane (HG, 3 a glutaraldehyde cross-linked bovine Type I collagen (BM, and 4 a resorbable polymer membrane (CP. Osteoblast differentiation was studied using an ALP assay with p-nitro phenyl phosphate as the substrate at 24 hours, 72 hours, and 1 week. A total of 50 µg/ml of EMD dissolved in 10 mM acetic acid was added into each well and the entire experimental protocol outlined above was repeated. Results: The osteoblast adhesion to collagen barriers showed a statistically insignificant reduction following the addition of EMD. Adhesion to the polymer barrier, although significantly lower when compared with collagen barriers, was unaffected by the addition of EMD. ALP activity after 1 week among the various groups was as follows: EMD alone (75.59±2

  4. Effects of Intercropping with Potato Onion on the Growth of Tomato and Rhizosphere Alkaline Phosphatase Genes Diversity

    Wu, Xia; Wu, Fengzhi; Zhou, Xingang; Fu, Xuepeng; Tao, Yue; Xu, Weihui; Pan, Kai; Liu, Shouwei


    Background and Aims: In China, excessive fertilization has resulted in phosphorus (P) accumulation in most greenhouse soils. Intercropping can improve the efficiency of nutrient utilization in crop production. In this study, pot experiments were performed to investigate the effects of intercropping with potato onion (Allium cepa L. var. aggregatum G. Don) on tomato (Solanum lycopersicum L.) seedlings growth and P uptake, the diversity of rhizosphere phosphobacteria and alkaline phosphatase (ALP) genes in phosphorus-rich soil. Methods: The experiment included three treatments, namely tomato monoculture (TM), potato onion monoculture (OM), and tomato/potato onion intercropping (TI-tomato intercropping and OI-potato onion intercropping). The growth and P uptake of tomato and potato onion seedlings were evaluated. The dilution plating method was used to determine the population of phosphate-solubilizing bacteria (PSB) and phosphate-mineralizing bacteria (PMB). The genomic DNAs of PSB and PMB in the rhizosphere of tomato and potato onions were extracted and purified, and then, with the primer set of 338f /518r, the PCR amplification of partial bacterial 16S rDNA sequence was performed and sequenced to determine the diversities of PSB and PMB. After extracting the total genomic DNAs from the rhizosphere, the copy numbers and diversities of ALP genes were investigated using real-time PCR and PCR-DGGE, respectively. Results: Intercropping with potato onion promoted the growth and P uptake of tomato seedlings, but inhibited those of potato onion. After 37 days of transplanting, compared to the rhizosphere of TM, the soil pH increased, while the electrolytic conductivity and Olsen P content decreased (p < 0.05) in the rhizosphere of TI. The populations and diversities of PSB, PMB, and ALP genes increased significantly in the rhizosphere of TI, compared to the rhizosphere of TM. Conclusion: The results indicated that intercropping with potato onion promoted the growth and P

  5. Patient pools and the use of "patient means" are valuable tools in quality control illustrated by a bone-specific alkaline phosphatase assay

    Hinge, Maja; Lund, Erik D.; Brandslund, Ivan;


    BACKGROUND: Quality control (QC) is an essential part of clinical biochemistry to ensure that laboratory test results are reliable and correct. Those tests without a defined reference method constitute a special challenge, as is the case with bone-specific alkaline phosphatase (BAP). METHODS...... AND RESULTS: The present study reports an example where a shift in a BAP assay was detected by use of a patient pool and supported by a retrospective calculation of "patient mean", while the external QC and specific assay control material were unaffected by the shift. CONCLUSIONS: Patient pools and the use...

  6. Synthesis of benzofuran derivatives as selective inhibitors of tissue-nonspecific alkaline phosphatase: effects on cell toxicity and osteoblast-induced mineralization.

    Marquès, Stéphanie; Buchet, René; Popowycz, Florence; Lemaire, Marc; Mebarek, Saïda


    Tissue-nonspecific alkaline phosphatase (TNAP) by hydrolyzing pyrophosphate, an inhibitor of apatite formation, promotes extracellular matrix calcification during bone formation and growth, as well as during ectopic calcification under pathological conditions. TNAP is a target for the treatment of soft tissue pathological ossification. We synthesized a series of benzofuran derivatives. Among these, SMA14, displayed TNAP activity better than levamisole. SMA14 was found to be not toxic at doses of up to 40μM in osteoblast-like Saos-2 cells and primary osteoblasts. As probed by Alizarin Red staining, this compound inhibited mineral formation in murine primary osteoblast and in osteoblast-like Saos-2 cells.

  7. The evaluation of 25-hydroxy vitamin D, calcium, phosphate and alkaline phosphatase levels in epileptic children under antiepileptic medication

    Keyhani doost Z


    Full Text Available "n 800x600 Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 st1":*{behavior:url(#ieooui } /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.0pt; font-family:"Times New Roman","serif";} Background: Epilepsy is a common disease in the pediatric neurology. There are frequent anti-epileptic drugs which are used in management of epilepsy. Anti-epileptic drugs may have some complications on bone and vitamin-D metabolism. In this study we aimed to evaluate vitamin-D metabolism in epileptic children."n"nMethods: The study was a prospective and cross sectional one. A total 89 epileptic children who were taking anti-epileptic drugs for longer than six months with no underlying disorder in Imam Khomeini and Bahrami Hospitals in Tehran, Iran were enrolled in our study"n"nResults: Forty nine boys and 40 girls were enrolled in this study; mean age of the patients was 7.8±2.1 years. Mean duration of anti-epileptic drug therapy was 2.3 years (SD=0.4, 70 of patients were under monotherapy and 19 were under polytherapy. None of the patients had signs of rickets. Serum calcium and phosphor levels were within normal ranges. Serum alkaline phosphates levels were increased more than two times in 43%. 42% had vitamin-D deficiency (25-OH Vit D<10 ng/ml and another 33% had vitamin-D insufficiency (10<25-oh Vit D<20 ng/ml. 29 patients (32% were taking prophylactic supplemental Vit D (200-400 IU/day. There was significant difference between patients taking supplemental vitamin-D as prophylaxis and patients who did not (p=0.04. There was no significant difference in vitamin-D levels between patients according to age, gender or different drugs."n"nConclusion: Periodic

  8. Marked Transient Alkaline Phosphatemia Following Pediatric Liver Transplantation

    Koneru, Baburao; Carone, Eduardo; Malatack, J. Jeffrey; Esquivel, Carlos O.; Starzl, Thomas E.


    An isolated marked transient rise in serum alkaline phosphatase levels in otherwise healthy children is a well-documented occurrence. However, in children undergoing liver transplantation, elevated alkaline phosphatase values raise the possibility of biliary obstruction, rejection, or both. During a 6-year period, 6 of 278 children undergoing liver transplantation exhibited a similar phenomenon as an isolated abnormality. None had rejection, biliary obstruction, or other allograft dysfunction during a long follow-up. Eventually and without intervention, the alkaline phosphatase levels returned to normal. These instructive cases suggest that caution be used in advocating Invasive procedures if elevated alkaline phosphatase levels are an isolated abnormality, and close observation with noninvasive testing is recommended. PMID:2658549

  9. The synthesis of Phosphate-repressible alkaline phosphatase do not appear to be regulated by ambient pH in the filamentous mould Neurospora crassa

    Nozawa Sérgio R.


    Full Text Available In order to investigate further the adaptive response of moulds to ambient pH, we have measured by ELISA the pho-2-encoded Pi-repressible alkaline phosphatase synthesised by Neurospora crassa. We showed that the 74A and pho-2A strains of this mould secrete similar amounts of the pho-2-encoded enzyme irrespective of ambient pH, when both the preg and pgov genes are not functional, i.e., in strains nuc-2+ growing under Pi-starvation. This suggests that pho-2, which is responsive to Pi starvation via the action of genes nuc-2, preg, pgov and nuc-1, is not a gene responsive to ambient pH and that the differential glycosylation observed for the Pi-repressible alkaline phosphatase retained by the mycelium at pH 5.6 or secreted into the growth medium at pH 8.0 is the genetic response to ambient pH sensing in N. crassa.

  10. Treatment of PCR products with exonuclease I and heat-labile alkaline phosphatase improves the visibility of combined bisulfite restriction analysis

    Watanabe, Kousuke; Emoto, Noriko; Sunohara, Mitsuhiro; Kawakami, Masanori; Kage, Hidenori; Nagase, Takahide; Ohishi, Nobuya [Department of Respiratory Medicine, The University of Tokyo Hospital, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan); Takai, Daiya, E-mail: [Department of Clinical Laboratory, The University of Tokyo Hospital, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan)


    Research highlights: {yields} Incubating PCR products at a high temperature causes smears in gel electrophoresis. {yields} Smears interfere with the interpretation of methylation analysis using COBRA. {yields} Treatment with exonuclease I and heat-labile alkaline phosphatase eliminates smears. {yields} The elimination of smears improves the visibility of COBRA. -- Abstract: DNA methylation plays a vital role in the regulation of gene expression. Abnormal promoter hypermethylation is an important mechanism of inactivating tumor suppressor genes in human cancers. Combined bisulfite restriction analysis (COBRA) is a widely used method for identifying the DNA methylation of specific CpG sites. Here, we report that exonuclease I and heat-labile alkaline phosphatase can be used for PCR purification for COBRA, improving the visibility of gel electrophoresis after restriction digestion. This improvement is observed when restriction digestion is performed at a high temperature, such as 60 {sup o}C or 65 {sup o}C, with BstUI and TaqI, respectively. This simple method can be applied instead of DNA purification using spin columns or phenol/chloroform extraction. It can also be applied to other situations when PCR products are digested by thermophile-derived restriction enzymes, such as PCR restriction fragment length polymorphism (RFLP) analysis.

  11. Inhibition of Alkaline Phosphatase from Pearl Oyster Pinctada fucata by o-Phthalaldehyde: Involvement of Lysine and Histidine Residues at the Active Site

    CHEN Hongtao; XIE Liping; YU Zhenyan; ZHANG Rongqing


    Alkaline phosphatase from Pinctada fucata was inactivated by o-phthalaldehyde (OPA). The inactivation followed pseudo first-order kinetics with a second rate constant of 0.167 (mmol/L)-1·min-1 at pH 7.5 and 25°C. A Tsou's plot analysis showed that inactivation occurred upon formation of one isoindole group. The OPA-modified enzyme lost the ability to bind with the specific affinity column and the presence of substrates or competitive inhibitors protected the enzyme from inactivation. The results revealed that the OPA-reaction site was at the enzyme substrate binding site. Prior modification of the enzyme by lysine or histidine specific reagent abolished formation of the isoindole derivatives, suggesting that lysine and histidine residues were involved in the OPA-induced inactivation. Taken together, OPA inactivated the alkaline phosphatase from Pinctada fucata by cross-linking lysine and histidine residues at the active site and formed an isoindole group at the substrate binding site of the enzyme.

  12. Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma

    Brasso, Klaus; Christensen, Ib Jarle; Johansen, Julia S;


    To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC).......To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC)....

  13. Age- and diapause-related acid and alkaline phosphatase activities in the intestine and malpighian tubules of the Colorado potato beetle, Leptinotarsa decemlineata (Say).

    Yi, S X; Adams, T S


    Specific activities for soluble (s) and membrane (m)-bound acid (ACP) and alkaline phosphatases (ALP) were determined in the midgut, hindgut, and Malpighian tubules for developing, prediapausing, and diapausing adult Colorado potato beetles, Leptinotarsa decemlineata (Say). High ACP activities were found in the hindgut and Malpighian tubules while high ALP activities were found in the Malpighian tubules. Variation in both ACP and ALP activities in each tissue reflects fluctuation in protein synthesis and secretion involved with digestion, excretion, and other unknown functions. Phosphatase activities in the tissues examined show the dynamic nature of diapause in this insect. Diapausing beetles showed increases in phosphatase activity after hormone treatments. JHA treatments increased s-ACP and m-ACP activities in all tissues but 20-HE did not increase activity in any tissue. Allatotropin tended to mimic the effects of JHA treatment. The s-ALP activity was also increased in all tissues whereas m-ALP was increased in the midgut and hindgut by JHA treatment. Malpighian tubule m-ALP activity was only increased by 20-HE treatments. Allatotropin was not as effective in increasing ALP activities as it was with ACP activities.

  14. A phage-displayed chicken single-chain antibody fused to alkaline phosphatase detects Fusarium pathogens and their presence in cereal grains

    Hu, Zu-Quan [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Li, He-Ping [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Zhang, Jing-Bo [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Huang, Tao [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Liu, Jin-Long; Xue, Sheng [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Wu, Ai-Bo [Institute for Agri-food Standards and Testing Technology, Laboratory of Quality and Safety Risk Assessment for Agro-products, Ministry of Agriculture, Shanghai Academy of Agricultural Sciences, 1000 Jinqi Road, Shanghai 201403 (China); Liao, Yu-Cai, E-mail: [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); National Center of Plant Gene Research, Wuhan 430070 (China)


    Graphical abstract: A phage-displayed chicken scFv antibody, FvSG7, binds on the surface antigen of conidiospores and the mycelia of F. verticillioides. Its fusion with alkaline phosphatase (AP) through a 218 linker displayed a 4-fold higher affinity compared with the parent scFv antibody and efficiently detected toxigenic Fusarium pathogens in cereal grains. Highlights: ► Generation of a highly reactive scFv antibody against F. verticillioides. ► Localization of the antibody binding to the surface target of F. verticillioides. ► Expression of the antibody–alkaline phosphatase (AP) fusion linked by a 218 linker. ► The antibody–AP fusion has a higher affinity than the parental antibody. ► The antibody–AP fusion detects toxigenic Fusarium pathogens in cereal grains. -- Abstract: Fusarium and its poisonous mycotoxins are distributed worldwide and are of particular interest in agriculture and food safety. A simple analytical method to detect pathogens is essential for forecasting diseases and controlling mycotoxins. This article describes a proposed method for convenient and sensitive detection of Fusarium pathogens that uses the fusion of single-chain variable fragment (scFv) and alkaline phosphatase (AP). A highly reactive scFv antibody specific to soluble cell wall-bound proteins (SCWPs) of F. verticillioides was selected from an immunized chicken phagemid library by phage display. The antibody was verified to bind on the surface of ungerminated conidiospores and mycelia of F. verticillioides. The scFv–AP fusion was constructed, and soluble expression in bacteria was confirmed. Both the antibody properties and enzymatic activity were retained, and the antigen-binding capacity of the fusion was enhanced by the addition of a linker. Surface plasmon resonance measurements confirmed that the fusion displayed 4-fold higher affinity compared with the fusion's parental scFv antibody. Immunoblot analyses showed that the fusion had good binding

  15. Aspartic acid-484 of nascent placental alkaline phosphatase condenses with a phosphatidylinositol glycan to become the carboxyl terminus of the mature enzyme.

    Micanovic, R; Bailey, C A; Brink, L; Gerber, L; Pan, Y C; Hulmes, J D; Udenfriend, S


    A carboxyl-terminal chymotryptic peptide from mature human placental alkaline phosphatase was purified by HPLC and monitored by a specific RIA. Sequencing and amino acid assay showed that the carboxyl terminus of the peptide was aspartic acid, representing residue 484 of the proenzyme as deduced from the corresponding cDNA. Further analysis of the peptide showed it to be a peptidoglycan containing one residue of ethanolamine, one residue of glucosamine, and two residues of neutral hexose. The inositol glycan is apparently linked to the alpha carboxyl group of the aspartic acid through the ethanolamine. Location of the inositol glycan on Asp-484 of the proenzyme indicates that a 29-residue peptide is cleaved from the nascent protein during the post-translational condensation with the phosphatidylinositol-glycan. PMID:3422741

  16. Microchannel conductivity measurements in microchip for on line monitoring of dephosphorylation rates of organic phosphates using paramagnetic-beads linked alkaline phosphatase.

    Kechadi, Mohammed; Sotta, Bruno; Gamby, Jean


    This paper presents the use of polymer coated microelectrodes for the realtime conductivity monitoring in a microchannel photoablated through the polymer without contact. Based on this strategy, a small conductometry sensor has been developed to record in time conductivity variation when an enzymatic reaction occurs through the channel. The rate constant determination, k2, for the dephosphorylation of organic phosphate-alkaline phosphatase-superparamagnetic beads complex using chemically different substrates such as adenosine monoesterphosphate, adenosine diphosphate and adenosine triphosphate was taken as an example to demonstrate selectivity and sensivity of the detection scheme. The k2 value measured for each adenosine phosphate decreases from 39 to 30 s(-1) in proportion with the number (3, 2 and 1) of attached phosphate moiety, thus emphasizing the steric hindrance effect on kinetics.

  17. Electron micrographic study of precipitates formed by interaction of silicic acid and alkaline phosphatase: contribution to a study of silica urolithiasis in cattle.

    Bailey, C B; Cheng, K J; Costerton, J W


    Association of alkaline phosphatase with silicic acid in precipitates formed in dilute solution was studied as a model for the nonspecific reaction between silicic acid and protein. Precipitates contained 68-83% of the silicic acid and 52-83% of the enzyme in the original mixture and were in the form of aggregates of roundish particles 150-800 nm in diameter. Enzyme protein formed a tightly bound layer on the surface of particles formed in solutions of freshly prepared silicic acid. The similarity between the ultrastructural features of precipitates from solutions of silicic acid and of internal portions of siliceous urinary calculi from cattle suggests that deposition of silica during development of such calculi is due, at least in part, to the interaction of protein with silicic acid in urine.

  18. Lactoferrin up-regulates intestinal gene expression of brain-derived neurotrophic factors BDNF, UCHL1 and alkaline phosphatase activity to alleviate early weaning diarrhea in postnatal piglets.

    Yang, Changwei; Zhu, Xi; Liu, Ni; Chen, Yue; Gan, Hexia; Troy, Frederic A; Wang, Bing


    The molecular mechanisms underlying how dietary lactoferrin (Lf) impacts gut development and maturation and protects against early weaning diarrhea are not well understood. In this study, we supplemented postnatal piglets with an Lf at a dose level of 155 and 285 mg/kg/day from 3 to 38 days following birth. Our findings show that the high dose of Lf up-regulated messenger RNA expression levels of genes encoding brain-derived neurotrophic factor (BDNF) and ubiquitin carboxy-terminal hydrolase L1 (ubiquitin thiolesterase (UCHL1) and, to a lesser extent, glial cell line-derived neurotrophic factor, in the duodenum (Pintestinal alkaline phosphatase activity (Pbrain-microbe axis that has not been previously reported.

  19. Partial Enteral Nutrition Preserves Elements of Gut Barrier Function, Including Innate Immunity, Intestinal Alkaline Phosphatase (IAP) Level, and Intestinal Microbiota in Mice.

    Wan, Xiao; Bi, Jingcheng; Gao, Xuejin; Tian, Feng; Wang, Xinying; Li, Ning; Li, Jieshou


    Lack of enteral nutrition (EN) during parenteral nutrition (PN) leads to higher incidence of infection because of gut barrier dysfunction. However, the effects of partial EN on intestina linnate immunity, intestinal alkaline phosphatase (IAP) and microbiota remain unclear. The mice were randomized into six groups to receive either standard chow or isocaloric and isonitrogenous nutritional support with variable partial EN to PN ratios. Five days later, the mice were sacrificed and tissue samples were collected. Bacterial translocation, the levels of lysozyme, mucin 2 (MUC2), and IAP were analyzed. The composition of intestinal microbiota was analyzed by 16S rRNA pyrosequencing. Compared with chow, total parenteral nutrition (TPN) resulted in a dysfunctional mucosal barrier, as evidenced by increased bacterial translocation (p microbiota (p intestinal microbiota.

  20. Variation of Alkaline Phosphatase Activity in Sediments of Shrimp Culture Ponds and Its Relationship with the Contents of C, N and P

    SU Yuepeng; MA Shen; DONG Shuanglin


    Nine enclosures (5 m × 5 m) were built in a Fenneropenaeus chinensis culture pond of Rushan Gulf in April, 2001.The probiotics and BIO ENERGIZER solution were applied for disparate treatments. Variations of alkaline phosphatase activity (APA) and its relationship with the contents of C, N and P in sediments were studied. Results show that APA of sediments increases from 3.096 nmol g-1min-1 to 5.407 nmol g-1nin-1 in culture period; the bacteria biomass is not the only factor to determine APA; the contents of total P and total organic carbon have a significant positive correlation with APA, while that of total nitrogen has a negative correlation. In addition, the contents of inorganic P and organic P are not regular with APA. By comparison, TOC shows a more significant coherence with APA, meaning that organic pollution in sediments affects APA remarkably.

  1. Multiwalled carbon nanotube modified screen-printed electrodes for the detection of p-aminophenol: Optimisation and application in alkaline phosphatase-based assays

    Lamas-Ardisana, Pedro Jose [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain); Queipo, Paula [Departamento de Fisica, Universidad de Oviedo, 33007 Oviedo, Asturias (Spain); Fanjul-Bolado, Pablo [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain); Costa-Garcia, Agustin [Departamento de Quimica Fisica y Analitica, Universidad de Oviedo, 33006 Oviedo, Asturias (Spain)], E-mail:


    Carboxylated multiwalled carbon nanotubes (MWCNT-COOH) were used to modify the working electrode surface of different screen-printed electrodes. The effect of this modification on the electrodic characteristics (double layer capacitance, electroactive area and heterogeneous rate constants for the electron transfer) was evaluated and optimized for the cyclic voltammetric determination of p-aminophenol. The enzymatic hydrolysis of p-aminophenylphosphate was employed for the quantification of alkaline phosphatase, one of the most important label enzymes in immunoassays. Finally, ELISA assays were carried out to quantify pneumolysin using this enzymatic system. Results obtained indicated that low superficial densities of MWCNT-COOH (0.03-0.06 {mu}g mm{sup -2}) yielded the same electrodic improvements but with better analytical properties.

  2. A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

    Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario


    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders.

  3. Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma

    Brasso, Klaus; Christensen, Ib Jarle; Johansen, Julia S.;


    BACKGROUND: To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC). METHODS: The biomarkers were determined by ELISAs in 153 metastatic PC patients before treatment with parenteral estrogen or total androgen...

  4. Calcium-phosphate biomineralization induced by alkaline phosphatase activity in Escherichia coli: localization, kinetics and potential signatures in the fossil record

    Cosmidis, Julie; Benzerara, Karim; Guyot, François; Skouri-Panet, Fériel; Duprat, Elodie; Férard, Céline; Guigner, Jean-Michel; Babonneau, Florence; Coelho, Cristina


    Bacteria are thought to play an important role in the formation of calcium-phosphate minerals composing marine phosphorites, as supported by the common occurrence of fossil microbes in these rocks. Phosphatase enzymes may play a key role in this process. Indeed, they may increase the supersaturation with respect to Ca-phosphates by releasing orthophosphate ions following hydrolysis of organic phosphorus. However, several questions remain unanswered about the cellular-level mechanisms involved in this model, and its potential signatures in the mineral products. We studied Ca-phosphate precipitation by different strains of Escherichia coli which were genetically modified to differ in the abundance and cellular localization of the alkaline phosphatase (PHO A) produced. The mineral precipitated by either E. coli or purified PHO A was invariably identified as a carbonate-free non-stoichiometric hydroxyapatite. However, the bacterial precipitates could be discriminated from the ones formed by purified PHO A at the nano-scale. PHO A localization was shown to influence the pattern of Ca-phosphate nucleation and growth. Finally, the rate of calcification was proved to be consistent with the PHO A enzyme kinetics. Overall, this study provides mechanistic keys to better understand phosphogenesis in the environment, and experimental references to better interpret the microbial fossil record in phosphorites.

  5. Enzymatic methods for the determination of pollution in seawater using salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius.

    Menzorova, Natalie I; Seitkalieva, Alexandra V; Rasskazov, Valerу A


    A new salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius (StAP) has been shown to have a unique property to hydrolyze substrate in seawater without loss of enzymatic activity. The enzyme has pH optimum at 8.0-8.5. Model experiments showed various concentrations of copper, zinc, cadmium and lead added to seawater or a standard buffer mixture to inhibit completely the enzyme activity at the concentrations of 15-150 μg/l. StAP sensitivity to the presence in seawater of metals, pesticides, detergents and oil products appears to be considerably less. Samples of seawater taken from aquatic areas of the Troitsy Bay of the Peter the Great Bay, Japan Sea have been shown to inhibit the enzyme activity; the same was shown for the samples of fresh waters. The phosphatase inhibition assay developed proved to be highly sensitive, technically easy-to use allowing to test a great number of samples.

  6. Loss of skeletal mineralization by the simultaneous ablation of PHOSPHO1 and alkaline phosphatase function: a unified model of the mechanisms of initiation of skeletal calcification.

    Yadav, Manisha C; Simão, Ana Maria Sper; Narisawa, Sonoko; Huesa, Carmen; McKee, Marc D; Farquharson, Colin; Millán, José Luis


    Endochondral ossification is a carefully orchestrated process mediated by promoters and inhibitors of mineralization. Phosphatases are implicated, but their identities and functions remain unclear. Alkaline phosphatase (TNAP) plays a crucial role promoting mineralization of the extracellular matrix by restricting the concentration of the calcification inhibitor inorganic pyrophosphate (PP(i)). Mutations in the TNAP gene cause hypophosphatasia, a heritable form of rickets and osteomalacia. Here we show that PHOSPHO1, a phosphatase with specificity for phosphoethanolamine and phosphocholine, plays a functional role in the initiation of calcification and that ablation of PHOSPHO1 and TNAP function prevents skeletal mineralization. Phospho1(-/-) mice display growth plate abnormalities, spontaneous fractures, bowed long bones, osteomalacia, and scoliosis in early life. Primary cultures of Phospho1(-/-) tibial growth plate chondrocytes and chondrocyte-derived matrix vesicles (MVs) show reduced mineralizing ability, and plasma samples from Phospho1(-/-) mice show reduced levels of TNAP and elevated plasma PP(i) concentrations. However, transgenic overexpression of TNAP does not correct the bone phenotype in Phospho1(-/-) mice despite normalization of their plasma PP(i) levels. In contrast, double ablation of PHOSPHO1 and TNAP function leads to the complete absence of skeletal mineralization and perinatal lethality. We conclude that PHOSPHO1 has a nonredundant functional role during endochondral ossification, and based on these data and a review of the current literature, we propose an inclusive model of skeletal calcification that involves intravesicular PHOSPHO1 function and P(i) influx into MVs in the initiation of mineralization and the functions of TNAP, nucleotide pyrophosphatase phosphodiesterase-1, and collagen in the extravesicular progression of mineralization.

  7. A rapid lateral flow immunoassay for the detection of tyrosine phosphatase-like protein IA-2 autoantibodies in human serum.

    Ingrid Kikkas

    Full Text Available Type 1 diabetes (T1D results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is subsequently captured on the anti-HA-Tag antibody-coated test line on the strip. The other site of the IA-2As is bound to biotinylated IA-2, allowing the complex to be visualized using colloidal gold nanoparticle-conjugated streptavidin. For this study, 35 serum samples from T1D patients and 44 control sera from non-diabetic individuals were analyzed with our novel assay and the results were correlated with two IA-2A ELISAs. Among the 35 serum samples from T1D patients, the IA-2A LFIA, the in-house IA-2A ELISA and the commercial IA-2A ELISA identified as positive 21, 29 and 30 IA-2A-positive sera, respectively. The major advantages of the IA-2A LFIA are its rapidity and simplicity.

  8. Targeting the active site of the placental isozyme of alkaline phosphatase by phage-displayed scFv antibodies selected by a specific uncompetitive inhibitor

    Kala Mrinalini


    Full Text Available Abstract Background The isozymes of alkaline phosphatase, the tissue non-specific, intestinal and placental, have similar properties and a high degree of identity. The placental isozyme (PLAP is an oncofetal antigen expressed in several malignancies including choriocarcinoma, seminoma and ovarian carcinoma. We had earlier attempted to isolate PLAP-specific scFv from a synthetic human immunoglobulin library but were unable to do so, presumably because of the similarity between the isozymes. In this work, we have employed a PLAP-specific uncompetitive inhibitor, L-Phe-Gly-Gly, to select isozyme specific scFvs. An uncompetitive inhibitor binds to the enzyme in the presence of substrate and stabilizes the enzyme-substrate complex. Several uncompetitive inhibitors have varying degrees of isozyme specificity for human alkaline phosphatase isozymes. A specific uncompetitive inhibitor would be able to unmask conformational differences between the otherwise very similar molecules. Also, such inhibitors would be directed to regions at/close to the active site of the enzyme. In this work, the library was first incubated with PLAP and the bound clones then eluted by incubation with L-Phe-Gly-Gly along with the substrate, para-nitro phenyl phosphate (pNPP. The scFvs were then studied with regard to the biochemical modulation of their binding, isozyme specificity and effect on enzyme activity. Results Of 13 clones studied initially, the binding of 9 was inhibited by L-Phe-Gly-Gly (with pNPP and 2 clones were inhibited by pNPP alone. Two clones had absolute and 2 clones had partial specificity to PLAP. Two clones were cross-reactive with only one other isozyme. Three scFv clones, having an accessible His6-tag, were purified and studied for their modulation of enzyme activity. All the three scFvs inhibited PLAP activity with the kinetics of competitive inhibition. Cell ELISA could demonstrate binding of the specific scFvs to the cell surface expressed PLAP

  9. Arbuscular mycorrhizal fungal diversity, root colonization, and soil alkaline phosphatase activity in response to maize-wheat rotation and no-tillage in North China.

    Hu, Junli; Yang, Anna; Zhu, Anning; Wang, Junhua; Dai, Jue; Wong, Ming Hung; Lin, Xiangui


    Monitoring the effects of no-tillage (NT) in comparison with conventional tillage (CT) on soil microbes could improve our understanding of soil biochemical processes and thus help us to develop sound management strategies. The objective of this study was to compare the species composition and ecological function of soil arbuscular mycorrhizal (AM) fungi during the growth and rotation of crops under NT and CT. From late June 2009 to early June 2010, 32 topsoil (0-15 cm) samples from four individual plots per treatment (CT and NT) were collected at both the jointing and maturation stages of maize (Zea mays L.) and wheat (Triticum aestivum L.) from a long-term experimental field that was established in an Aquic Inceptisol in North China in June 2006. The AM fungal spores were isolated and identified and then used to calculate species diversity indices, including the Shannon- Wiener index (H'), Evenness (E), and Simpson's index (D). The root mycorrhizal colonization and soil alkaline phosphatase activity were also determined. A total of 34 species of AM fungi within nine genera were recorded. Compared with NT, CT negatively affected the soil AM fungal community at the maize sowing stage, leading to decreases in the average diversity indices (from 2.12, 0.79, and 0.82 to 1.79, 0.72, and 0.74 for H', E, and D, respectively), root mycorrhizal colonization (from 28% to 20%), soil alkaline phosphatase activity (from 0.24 to 0.19 mg/g/24 h) and available phosphorus concentration (from 17.4 to 10.5 mg/kg) at the maize jointing stage. However, reductions in diversity indices of H', E, and D were restored to 2.20, 0.81, and 0.84, respectively, at the maize maturation stage. CT should affect the community again at the wheat sowing stage; however, a similar restoration in the species diversity of AM fungi was completed before the wheat jointing stage, and the highest Jaccard index (0.800) for similarity in the species composition of soil AM fungi between CT and NT was recorded at

  10. Detection of prostatic cancer by solid-phase radioimmunoassay of serum prostatic acid phosphatase. [/sup 125/I tracer technique

    Foti, A.; Cooper, J.F.; Herschman, H.; Malvaez, R.R.


    We compared our radioimmunoassay with the standard enzyme assay for prostatic acid phosphatase in the diagnosis of prostatic cancer. Serum samples from 50 controls, 113 patients with prostatic cancer, 36 with benign prostatic hyperplasia, 83 with other cancers, 20 with gastrointestinal disorders and 28 with total prostatectomies were randomized and studied by radioimmunoassay and enzyme assay. When the upper limit was set at 8.0 ng per milliliter (mean + 4 S.D.) the radioimmunoassay diagnosed prostatic cancer in 33, 79, 71 and 92 percent of the patients with Stage I, II, III and IV disease. In contrast, the enzyme assay detected elevations of enzyme in the serum of 12, 15, 29, and 60 percent respectively. No false-positive results were detected by either assay in normal controls but the radioimmunoassay test was positive in two patients with benign prostatic hyperplasia, in one patient after total prostatectomy, in nine with other cancers and in one of the group with gastrointestinal disorders. In contrast to the enzyme assay, the radioimmunoassay distinguished over half the cases of intracapsular prostatic cancer.

  11. Effects of Myriophyllum spicatum on Kinetic Characteristics of Alkaline Phosphatase in Water and Sediments%狐尾藻对水体和沉积物中碱性磷酸酶动力学特征的影响

    赵海超; 王圣瑞; 金相灿; 焦立新


    The enzyme mechanism of an inactivating eutrophic lake by submerged plants was determined by analyzing the kinetic parameter variations of alkaline phosphataae in the overlying water, interstitial water and sediments under indoor simulating conditions with the plant Myriophyllum spicatum. The results indicated that under the experimental condition Vmax of the alkaline phosphatase, the overlying water, the interstitial water and the sediments were all reduced by planting Myriophyllum spicatum. Myriophyllum spicatum inactivated the alkaline phosphatase in the overlying water and sediments obviously, and mainly inactivated the dissolved alkaline phosphatase Vmax in the interstitial water. The effect of Myriophyllum spicatum on alkaline phosphatase in the soil was higher than that in the sediment of same nutrition. The Vmax and Km of the alkaline phosphatase in the overlying water of the soil substrate were higher than those of the sediment substrate, and the Vmax and Km of the alkaline phosphatase in the soil were lower than those in the sediment. The Vmax of the alkaline phosphatase in the sediments ascended opposite to that in the overlying water. The alkaline phosphatase in the interstitial water had obvious seasonal variation and characteristics, and Vmax attained the highest value from July to August.%在室内模拟条件下栽培狐尾藻,通过对上覆水、间隙水和沉积物中碱性磷酸酶动力学参数变化的分析,揭示了沉水植物对湖泊富营养化影响的酶学机制.结果表明:在试验条件下,栽培狐尾藻使上覆水、间隙水和沉积物中的碱性磷酸酶的最大反应速率(Vmax)均有所降低;狐尾藻对上覆水和底质中碱性磷酸酶反应速率及亲和力的抑制作用比较明显,对间隙水主要是抑制溶解性碱性磷酸酶的Vmax;狐尾藻对土壤中碱性磷酸酶的影响比同一营养水平的沉积物大,与沉积物相比,土壤作底质时上覆水中碱性磷酸酶的Vmax和K(ms)(米氏

  12. Extracellular Matrix Proteins, Alkaline Phosphatase and Pyrophosphate as Molecular Determinants of Bone, Tooth, Kidney and Vascular Calcification

    McKee, Marc D.


    Progress in biomineralization research in recent years has identified, characterized and described functions for key noncollagenous extracellular matrix proteins regulating crystal growth in the skeleton and dentition. Some of these same proteins expressed in soft tissues undergoing pathologic calcification also inhibit ectopic crystal growth. In addition to extracellular matrix proteins regulating matrix mineralization, the enzyme tissue-nonspecific alkaline phosphatase—which is highly expressed by cells in mineralized tissues—cleaves pyrophosphate, an anionic small-molecule inhibitor of mineralization. Together with the required mineral ion availability necessary for crystal growth, these molecular determinants appear to function in limiting the spread of pathologic calcification seen in soft tissues such as blood vessels and kidneys. Osteopontin, in particular, is a potent calcification inhibitor that accumulates in mineralized tissues and in calcified deposits during vascular calcification and nephrolithiasis/urolithiasis. Additional research is required to establish the exact temporal sequence in which the molecular determinants of pathologic calcification appear relative to mineral crystal growth in different tissues, and to establish their relationship (if any) to the activation of osteogenic differentiation programs.

  13. Serum sialic acid and CEA concentrations in human breast cancer.

    Hogan-Ryan, A.; Fennelly, J J; Jones, M.; Cantwell, B; Duffy, M J


    The concentration of bound sialic acid in the sera of 56 normal subjects and 65 subjects with breast cancer was measured, in order to determine (1) whether serum sialic acid concentrations are raised in breast cancer and (2) whether the concentration of sialic acid in serum reflects tumour stage. The amount of sialic acid in serum was compared to serum carcinoembryonic antigen (CEA) values. Urinary hydroxyproline and serum alkaline phosphatase concentrations were used as indicators of bone an...

  14. Prokaryotic responses to ammonium and organic carbon reveal alternative CO2 fixation pathways and importance of alkaline phosphatase in the mesopelagic North Atlantic

    Federico Baltar


    Full Text Available To decipher the response of mesopelagic prokaryotic communities to input of nutrients, we tracked changes in prokaryotic abundance, extracellular enzymatic activities, heterotrophic production, dark dissolved inorganic carbon (DIC fixation, community composition (16S rRNA sequencing and community gene expression (metatranscriptomics in 3 microcosm experiments with water from the mesopelagic North Atlantic. Responses in 3 different treatments amended with thiosulfate, ammonium or organic matter (i.e. pyruvate plus acetate were compared to unamended controls. The strongest stimulation was found in the organic matter enrichments, where all measured rates increased >10-fold. Strikingly, in the organic matter treatment, the dark DIC fixation rates —assumed to be related to autotrophic metabolisms— were equally stimulated as all the other heterotrophic-related parameters. This increase in DIC fixation rates was paralleled by an up-regulation of genes involved in DIC assimilation via anaplerotic pathways. Alkaline phosphatase was the metabolic rate most strongly stimulated and its activity seemed to be related to cross-activation by nonpartner histidine kinases, and/or the activation of genes involved in the regulation of elemental balance during catabolic processes. These findings suggest that episodic events such as strong sedimentation of organic matter into the mesopelagic might trigger rapid increases of originally rare members of the prokaryotic community, enhancing heterotrophic and autotrophic carbon uptake rates, ultimately affecting carbon cycling. Our experiments highlight a number of fairly unstudied microbial processes of potential importance in mesopelagic waters that require future attention.

  15. A fluorometric assay for alkaline phosphatase activity based on β-cyclodextrin-modified carbon quantum dots through host-guest recognition.

    Tang, Cong; Qian, Zhaosheng; Huang, Yuanyuan; Xu, Jiamin; Ao, Hang; Zhao, Meizhi; Zhou, Jin; Chen, Jianrong; Feng, Hui


    A convenient, reliable and highly sensitive assay for alkaline phosphatase (ALP) activity in the real-time manner is developed based on β-cyclodextrin-modified carbon quantum dots (β-CD-CQDs) nanoprobe through specific host-guest recognition. Carbon quantum dots were first functionalized with 3-aminophenyl boronic acid to produce boronic acid-functionalized CQDs, and then further modified with hydropropyl β-cyclodextrins (β-CD) through B-O bonds to form β-CD-CQDs nanoprobe. p-Nitrophenol phosphate disodium salt is used as the substrate of ALP, and can hydrolyze to p-nitrophenol under the catalysis of ALP. The resulting p-nitrophenol can enter the cavity of β-CD moiety in the nanoprobe due to their specific host-guest recognition, where photoinduced electron transfer process between p-nitrophenol and CQDs takes place to efficiently quench the fluorescence of the probe. The correlation between quenched fluorescence and ALP level can be used to establish quantitative evaluation of ALP activity in a broad range from 3.4 to 100.0U/L with the detection limit of 0.9U/L. This assay shows a high sensitivity to ALP even in the presence of a very high concentration of glucose. This study demonstrates a good electron donor/acceptor pair, which can be used to design general detection strategy through PET process, and also broadens the application of host-guest recognition for enzymes detection in clinical practice.

  16. Evaluation of Anti-aging Compounds Using the Promoters of Elastin and Fibrillin-1 Genes Combined with a Secreted Alkaline Phosphatase Reporter in Normal Human Fibroblasts.

    Lin, Chih-Chien; Yang, Chao-Hsun; Kuo, Wan-Ting; Chen, Cheng-Yu


    Elastic fibers are major constituents of the extracellular matrix (ECM) in dynamic tissues in the human body, and regulation of elastin and fibrillin-1 expression mediates the formation of these fibers. Traditional assays for the measurement of elastin and fibrillin-1, such as western blotting, Luna staining and immunostaining, are relatively complex and time-consuming. Thus, a relatively simple assay system that also provides rational results is urgently needed. In the study, we aimed to develop a human cell-based assay system that can be used to analyze functional compounds using the promoters of elastin (ELN) and fibrillin-1 (FBN1) genes integrated with a secreted alkaline phosphatase (SEAP) reporter in normal human fibroblast cells. We used this system to assess anti-aging compounds. We used several regulators of elastinogenesis, including retinol, coenzyme Q10, deoxyArbutin and Elestan(TM) (Manilkara multinervis leaf extract), to verify the efficacy of this assay system. Our results demonstrate that this assay system can be used as a fast and realistic method for identifying anti-aging components for future use in foods, cosmetics and drugs.

  17. Naked-eye sensitive detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) based on a horseradish peroxidase catalytic colorimetric system with Cu(ii).

    Shi, Dongmin; Sun, Yue; Lin, Lin; Shi, Chunjun; Wang, Guangfeng; Zhang, Xiaojun


    In this paper, a novel colorimetric method for the detection of alkaline phosphatase (ALP) and pyrophosphate (PPi) was designed based on a Cu(2+)-horseradish peroxidase (HRP)-3,3',5,5'-tetra-methylbenzidine (TMB)-H2O2 system. In the presence of ALP, l-ascorbic acid-2-phosphate (AAP) could be hydrolyzed to ascorbic acid which could reduce Cu(2+) to Cu(+) to inhibit the enzymatic activity of HRP in the colorimetric system. The change in absorbance was found to be proportional to the ALP concentration with a linear detection range and a limit of detection of 5.4 mU mL(-1). In the presence of PPi, because Cu(2+) was chelated by PPi, the conversion of Cu(ii) by AA was effectively inhibited. The color of the HRP-TMB-H2O2 system with Cu(2+) showed blue. The HRP-TMB-H2O2 system with the Cu(2+) colorimetric system could also detect PPi with a satisfying result. In summary, this method possesses sensitivity, reproducibility, and cost-effectiveness without labelling and separation and the use of a colorimetric method is more in line with the requirements of on-site detection and green chemistry.

  18. Copper(II) complexes with cyanoguanidine and o-phenanthroline: Theoretical studies, in vitro antimicrobial activity and alkaline phosphatase inhibitory effect

    Martínez Medina, Juan J.; Islas, María S.; López Tévez, Libertad L.; Ferrer, Evelina G.; Okulik, Nora B.; Williams, Patricia A. M.


    Calculations based on density functional methods are carried out for two Cu(II) complexes with cyanoguanidine (cnge) and o-phenanthroline (o-phen): [Cu(o-phen)2(cnge)](NO3)2ṡ2H2O (1) and [Cu(o-phen)(cnge)(H2O)(NO3)2] (2). The calculated geometrical parameters are in agreement with the experimental values. The results of Atoms in Molecules (AIM) topological analysis of the electron density indicate that the Cu-N(phen) bonds in complex (1) have lower electron density, suggesting that those bonds are stronger in complex (2). Moreover, the ionic character of the Cu-N bond in the complex (1) is slightly stronger than that in the complex (2) and this situation would explain the fact that only complex (2) was stable in water solution. For this reason, the antimicrobial and enzymatic assays were performed using complex (2). It is well known that the increased use of antibiotics has resulted in the development of resistant bacterial and fungal strains. In this context, the study of novel antimicrobial agents has an enormous importance and metal complexes represent an interesting alternative for the treatment of infectious diseases. The aim of this work is to prove the modification of some biological properties like antimicrobial activity or alkaline phosphatase inhibitory activity upon copper complexation.

  19. Alkaline phosphatases are involved in the response of Aedes aegypti larvae to intoxication with Bacillus thuringiensis subsp. israelensis Cry toxins.

    Stalinski, Renaud; Laporte, Frédéric; Després, Laurence; Tetreau, Guillaume


    Bacillus thuringiensis subsp. israelensis (Bti) is a natural pathogen of dipterans widely used as a biological insecticide for mosquito control. To characterize the response of mosquitoes to intoxication with Bti, the transcriptome profile of Bti-exposed susceptible Aedes aegypti larvae was analysed using Illumina RNA-seq. Gene expression of 11 alkaline phosphatases (ALPs) was further investigated by real time quantitative polymerase chain reaction and ALP activity was measured in the susceptible strain and in four strains resistant to a single Bti Cry toxin or to Bti. These strains were unexposed or exposed to their toxin of selection. Although all resistant strains constitutively exhibited a higher level of transcription of ALP genes than the susceptible strain, they showed a lower total ALP activity. The intoxication with different individual Cry toxins triggered a global pattern of ALP gene under-transcription in all the one-toxin-resistant strains but involving different specific sets of ALPs in each resistant phenotype. Most of the ALPs involved are not known Cry-binding proteins. RNA interference experiment demonstrated that reducing ALP expression conferred increased the survival of larvae exposed to Cry4Aa, confirming the involvement of ALP in Cry4Aa toxicity.

  20. Bifunctional coating based on carboxymethyl chitosan with stable conjugated alkaline phosphatase for inhibiting bacterial adhesion and promoting osteogenic differentiation on titanium

    Zheng, Dong; Neoh, Koon Gee; Kang, En-Tang


    In this work, alkaline phosphatase (ALP) was covalently immobilized on carboxymethyl chitosan (CMCS)-coated polydopamine (PDA)-functionalized Ti to achieve a bifunctional surface. Our results showed ∼89% reduction in Staphylococcus epidermidis adhesion on this surface compared to that on pristine Ti. The ALP-modified Ti supported cell proliferation, and significantly enhanced cellular ALP activity and calcium deposition of osteoblasts, human mesenchymal stem cells (hMSCs) and human adipose-derived stem cells (hADSCs). The extent of enhancement in the functions of these cells is dependent on the surface density of immobilized ALP. The substrate prepared using an ALP solution of 50 μg/cm2 resulted in 44%, 54% and 129% increase in calcium deposited by osteoblasts, hMSCs and hADSCs, respectively, compared to those cultured on pristine Ti. The ALP-modified substrates also promoted the osteogenic differentiation of hMSCs and hADSCs by up-regulating gene expressions of runt-related transcription factor 2 (RUNX2), osterix (OSX), and osteocalcin (OC) in the two types of stem cells. The surface-immobilized ALP was stable after being subjected to 1 h immersion in 70% ethanol and autoclaving at 121 °C for 20 min. However, the enzymatic bioactivity of the surface-immobilized ALP was reduced by about 50% after these substrates were immersed in phosphate buffered saline (PBS) or PBS containing lysozyme for 14 days.

  1. A single electrochemical biosensor for detecting the activity and inhibition of both protein kinase and alkaline phosphatase based on phosphate ions induced deposition of redox precipitates.

    Shen, Congcong; Li, Xiangzhi; Rasooly, Avraham; Guo, Linyan; Zhang, Kaina; Yang, Minghui


    Protein kinase (PKA) and alkaline phosphatase (ALP) are clinically relevant enzymes for a number of diseases. In this work, we developed a new simple electrochemical biosensor for the detection of the activity and inhibition of both PKA and ALP. One common feature of the PKA and ALP catalyzing process is that PKA can hydrolysis adenosine-5'-triphosphate (ATP) and ALP can hydrolysis pyrophosphate, both reactions produce phosphate ions, and the amount of phosphate ion produced is proportional to enzyme activity. Our assay is based on the principle that phosphate ions react with molybdate to form redox molybdophosphate precipitates on the electrode surface, thus generating electrochemical current. The detection limit for PKA and ALP were much lower than existing assays. The biosensor has good specificity and was used to measure drug-stimulated PKA from lysates of HeLa cells. We also evaluated the use of the biosensor as a screening tool for enzyme inhibitors. To the best of our knowledge, this is the first report of a biosensor capable of detecting the activity of both PKA and ALP. This tool has the potential to simplify PKA and ALP clinical measurement, thereby improving diagnostics of relevant diseases. It also may serve as the basis for a simple screening method for new enzyme inhibitors for disease treatment.

  2. The antioxidant effects of vitamin C on liver enzymes: aspartate aminotransferase, alanine aminotranferease, alkaline phosphatase and gamma-glutamyltransferase activities in rats under Paraquat insult

    Benjamin Nnamdi Okolonkwo


    Full Text Available Paraquat (PQ is a bipyridylium herbicide; applied around trees in orchards and between crop rows to control broad-leaved and grassy weeds. Its oxidation results in the formation of superoxides which causes damage to cellular components. In this study, we determined the antioxidant effect vitamin C has on the liver enzymes [aspartate aminotransferase (SGOT, alanine aminotranferease (SGPT, alkaline phosphatase (ALP, and gamma-glutamyltransferase (GGT] of rats under this toxic insult. Male rats in groups (A, B, C and D were intraperitoneally injected with different sublethal increasing doses (0, 0.02, 0.04 and 0.06 g/kg body weigh of PQ respectively on monthly basis. Subsequently, the subgroups (A2, B2, C2 and D2 were given orally, 200 mg/L vitamin C, while the subgroups A1, B1, C1, and D1, received only water. Four animals per subgroup were decapitated on monthly basis and blood samples taken for enzyme assay. The parameters studied were - SGOT, SGPT, ALP and GGT - liver enzymes. The dose and time dependent PQ toxicity effect resulted in highly elevated Liver enzymes activities. The subgroups on vitamin C had significantly lower enzyme activities when compared to the same subgroups on only PQ insult. But the values were high when compared to the control subgroups (A1 and A2. These results were indication that vitamin C when given at moderate doses and maintained for a longer period could be a life saving adjunct to toxic insult.

  3. Uptake of nickel from 316L stainless steel into contacting osteoblastic cells and metal ion interference with BMP-2-induced alkaline phosphatase.

    Mölders, Martina; Felix, Joachim; Bingmann, Dieter; Hirner, Alfred; Wiemann, Martin


    Bone cells contacting nickel (Ni)-containing implant materials may be affected by Ni species via disturbed signaling pathways involved in bone cell development. Here we analyze effects of the Ni-containing steel 316L and major metal constituents thereof on bone morphogenetic protein-2 (BMP-2)-induced alkaline phosphatase (ALP) of MC3T3-E1 cells. While cells grew normally on 316L, cellular Ni content increased 10-fold vs. control within 4 days. With respect to the major components of 316L, Ni2+ (3-50 microM) was most inhibitory to BMP-2-induced ALP, whereas even 50 microM Fe3+, Cr3+, Mo5+, or Mn2+ had no such effect. In line with this, BMP-2-induced ALP was significantly reduced in cells on 316L. This effect was not prevented by the metal ion chelator diethylenetriaminepentaacetic acid (DTPA). Instead, DTPA abolished the stimulatory effect of BMP-2 on ALP, pointing to chelatable metal ions involved. Zn2+, as one possible candidate, antagonized the Ni2+ inhibition of BMP-2-induced ALP in both MC3T3-E1 and human bone marrow stromal cells. Results show that cells contacting 316L steel are exposed to increased concentrations of Ni which suffice to impair BMP-2-induced ALP activity. Zn2+, as a competitor of this inhibition, may help to restore normal osteoblastic function and bone development under these conditions.

  4. Development of a nanobody-alkaline phosphatase fusion protein and its application in a highly sensitive direct competitive fluorescence enzyme immunoassay for detection of ochratoxin A in cereal.

    Liu, Xing; Xu, Yang; Wan, De-bin; Xiong, Yong-hua; He, Zhen-yun; Wang, Xian-xian; Gee, Shirley J; Ryu, Dojin; Hammock, Bruce D


    A rapid and sensitive direct competitive fluorescence enzyme immunoassay (dc-FEIA) for ochratoxin A (OTA) based on a nanobody (Nb)-alkaline phosphatase (AP) fusion protein was developed. The VHH (variable domain of heavy chain antibody) gene of Nb28 was subcloned into the expression vector pecan45 containing the AP double-mutant gene. The Nb28-AP construct was transformed into Escherichia coli BL21(DE3)plysS, and soluble expression in bacteria was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. Both the Nb properties and AP enzymatic activity were validated by colorimetric and fluorometric analysis. The 50% inhibitory concentration and the detection limit of the dc-FEIA were 0.13 and 0.04 ng/mL, respectively, with a linear range of 0.06-0.43 ng/mL. This assay was compared with LC-MS/MS, and the results indicated the reliability of Nb-AP fusion protein-based dc-FEIA for monitoring OTA contamination in cereal.

  5. High frequencies of elevated alkaline phosphatase activity and rickets exist in extremely low birth weight infants despite current nutritional support

    Parker Bruce R


    Full Text Available Abstract Background Osteopenia and rickets are common among extremely low birth weight infants (ELBW, Methods We evaluated all ELBW infants admitted to Texas Children's Hospital NICU in 2006 and 2007. Of 211 admissions, we excluded 98 patients who were admitted at >30 days of age or did not survive/stay for >6 weeks. Bone radiographs obtained in 32 infants were reviewed by a radiologist masked to laboratory values. Results In this cohort of 113 infants, P-APA was found to have a significant inverse relationship with BW, gestational age and serum phosphorus. In paired comparisons, P-APA of infants Conclusion Elevation of P-APA >600 IU/L was very common in ELBW infants. BW was significantly inversely related to both P-APA and radiologic rickets. No single value of P-APA was related to radiological findings of rickets. Given the very high risk of osteopenia and rickets among ELBW infants, we recommend consideration of early screening and early mineral supplementation, especially among infants

  6. Conformational changes in the bilirubin-human serum albumin complex at extreme alkaline pH

    Honoré, B; Frandsen, P C


    Light-absorption, c.d. and fluorescence of the bilirubin-albumin complex were investigated at extreme alkaline pH. Above pH 11.1 albumin binds the bilirubin molecule, twisted oppositely to the configuration at more neutral pH. On the basis of light-absorption it is shown that two alkaline...... between tryptophan-214 and bilirubin, and partly exposing the liganded bilirubin to the solvent. Udgivelsesdato: 1986-Jun-1...

  7. 碱性磷酸酶试剂开瓶的稳定性研究%Study on stability of alkaline phosphatase reagent after the packaging was opened

    段爱军; 段爱华


    Objective:To study the stability of alkaline phosphatase(ALP) after the packaging was opened and to provide the data for purchasing right amount reageng. Methods:The method of synchronous measurement was used and five kits(S0, S1, S2, S3, S4) were chosen. ALP routine test was performed using the kit S4 at eight a.m. in first day. The remaining reagent was covered tightly and preserved at 2-8 ℃. Then S3, S2, S1 and S0 kits were opened at the same time in second, third, fourth and fifth day using the same method, respectively. Five of the same parameters for ALP detection channel (ALP0, ALP1, ALP2, ALP3, ALP4, corresponding to S0, S1, S2, S3, S4) were also set in the instrument according to the reagent instruction. The detection channels were calibrated using the calibration materials(BioSino Bio-technology and Science Inc) in Selectra-E fully automatic biochemical analyzer. Results: Compared with the results obtained by S0, the results from S3 and S4 showed statistically significance(P0.05). Conclusion: The ALP reagent is stable in 48 hours after the packaging was opened and the laboratory should choose suitable packing size of reagent kit according to the dosage to use.%目的:研究碱性磷酸酶(alkaline phosphatase, ALP)试剂开瓶后的稳定性,为实验室合理购买试剂包规格提供依据。方法:采用同步测量设计方案,首先选择5个试剂盒,分别记为S0、S1、S2、S3、S4,第1天上午8点启用试剂S4进行ALP常规检测,未用完试剂盖紧盖,置2~8℃保存备用;第2天、第3天、第4天上午相同时间、相同方式启用S3、S2、S1;第5天启用试剂S0;在仪器上按试剂说明书设置5个相同参数的ALP检测通道,记为ALP0、ALP1、ALP2、ALP3、ALP4,对应试剂为S0、S1、S2、S3、S4;在荷兰威图Selectra-E全自动生化仪上用校准品(中生北控生物科技有限公司)校准各个通道;测量样品并进行结果比较分析。结果:试剂 S3、S4与试剂 S0

  8. Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B{sub 1} detection in cereal

    Shu, Mei [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Xu, Yang, E-mail: [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Liu, Xing [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); College of Food Science and Technology, Hainan University, No. 58 Renmin Avenue, Haikou 570228 (China); Li, Yanping; He, Qinghua [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Tu, Zhui [State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Fu, Jinheng [Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047 (China); Gee, Shirley J.; Hammock, Bruce D. [Department of Entomology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616 (United States)


    A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB{sub 1} was developed. The anti-idiotypic nanobody–alkaline phosphatase (Ab2β−Nb−AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB{sub 1} were 2.69 and 0.35 ng mL{sup −1}, respectively, with a linear range of 0.93–7.73 ng mL{sup −1}. The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL{sup −1}, and the IC{sub 50} was 0.89 ± 0.09 ng mL{sup −1} with a linear range of 0.29–2.68 ng mL{sup −1}. Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β−Nb−AP fusion protein based one-step competitive immunoassay for monitoring FB{sub 1} contamination in cereals. The Ab2β−Nb−AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems. - Highlights: • Ab2β−Nb−AP has the potential to replace chemically-coupled probes. • Ab2β−Nb−AP is homogeneous enzyme-labelled antigen can be prepared reproducibly. • We developed a green and rapid one-step competitive enzyme immunoassay. • The sensitivity of one-step CLIA was 9-folds higher than two-step ELISA.

  9. Mechanical loading induced expression of bone morphogenetic protein-2,alkaline phosphatase activity,and collagen synthesis in osteoblastic MC3T3-E1 cells

    LU Hong-fei; MAI Zhi-hui; XU Ye; WANG Wei; AI Hong


    Background Bone morphogenetic protein(BMP)-2,alkaline phosphatase(ALP),and collagen typeⅠ?are known to play a critical role in the process of bone remodeling.However,the relationship between mechanical strain and the expression of BMP-2,ALP,and COL-Ⅰ?in osteoblasts was still unknown.The purpose of this study was to investigate the effects of different magnitudes of mechanical strain on osteoblast morphology and on the expression of BMP-2,ALP,and COL-Ⅰ.Methods Osteoblast-like cells were flexed at four deformation rates(0,6%,12%,and 18% elongation).The expression of BMP-2 mRNA,ALP,and COL-Ⅰ?in osteoblast-like cells were determined by real-time quantitative reverse transcription polymerase chain reaction,respectively.The results were subjected to analysis of variance(ANOVA)using SPSS 13.0 statistical software.Results The cells changed to fusiform and grew in the direction of the applied strain after the mechanical strain was loaded.Expression level of the BMP-2,ALP,and COL-Ⅰ?increased magnitude-dependently with mechanical loading in the experimental groups,and the 12% elongation group had the highest expression(P<0.05).Conclusion Mechanical strain can induce morphological change and a magnitude-dependent increase in the expression of BMP-2,ALP,and COL-Ⅰ?mRNA in osteoblast-like cells,which might influence bone remodeling in orthodontic treatment.

  10. Boron Induces Early Matrix Mineralization via Calcium Deposition and Elevation of Alkaline Phosphatase Activity in Differentiated Rat Bone Marrow Mesenchymal Stem Cells

    Bent-al-hoda Movahedi Najafabadi


    Full Text Available Objective: Boron (B is essential for plant development and might be an essential micronutrient for animals and humans. This study was conducted to characterize the impact of boric acid (BA on the cellular and molecular nature of differentiated rat bone marrow mesenchymal stem cells (BMSCs. Materials and Methods: In this experimental study, BMSCs were extracted and expanded to the 3rd passage, then cultured in Dulbecco’s Modified Eagle’s Medium (DMEM complemented with osteogenic media as well as 6 ng/ml and 6 μg/ml of BA. After 5, 10, 15 and 21 days the viability and the level of mineralization was determined using MTT assay and alizarin red respectively. In addition, the morphology, nuclear diameter and cytoplasmic area of the cells were studied with the help of fluorescent dye. The concentration of calcium, activity of alanine transaminase (ALT, aspartate transaminase (AST, lactate dehydrogenase (LDH and alkaline phosphatase (ALP as well as sodium and potassium levels were also evaluated using commercial kits and a flame photometer respectively. Results: Although 6 μg/ml of BA was found to be toxic, a concentration of 6 ng/ml increased the osteogenic ability of the cell significantly throughout the treatment. In addition it was observed that B treatment caused the early induction of matrix mineralization compared to controls. Conclusion: Although more investigation is required, we suggest the prescription of a very low concentration of B in the form of BA or foods containing BA, in groups at high risk of osteoporosis or in the case of bone fracture.

  11. Induction of bone-type alkaline phosphatase in human vascular smooth muscle cells: roles of tumor necrosis factor-alpha and oncostatin M derived from macrophages.

    Shioi, Atsushi; Katagi, Miwako; Okuno, Yasuhisa; Mori, Katsuhito; Jono, Shuichi; Koyama, Hidenori; Nishizawa, Yoshiki


    Inflammatory cells such as macrophages and T lymphocytes play an important role in vascular calcification associated with atherosclerosis and cardiac valvular disease. In particular, macrophages activated with cytokines derived from T lymphocytes such as interferon-gamma (IFN-gamma) may contribute to the development of vascular calcification. Moreover, we have shown the stimulatory effect of 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) on in vitro calcification through increasing the expression of alkaline phosphatase (ALP), an ectoenzyme indispensable for bone mineralization, in vascular smooth muscle cells. Therefore, we hypothesized that macrophages may induce calcifying phenotype, especially the expression of ALP in human vascular smooth muscle cells (HVSMCs) in the presence of IFN-gamma and 1,25(OH)2D3. To test this hypothesis, we used cocultures of HVSMCs with human monocytic cell line (THP-1) or peripheral blood monocytes (PBMCs) in the presence of IFN-gamma and 1,25(OH)2D3. THP-1 cells or PBMCs induced ALP activity and its gene expression in HVSMCs and the cells with high expression of ALP calcified their extracellular matrix by the addition of beta-glycerophosphate. Thermostability and immunoassay showed that ALP induced in HVSMCs was bone-specific enzyme. We further identified tumor necrosis factor-alpha (TNF-alpha) and oncostatin M (OSM) as major factors inducing ALP in HVSMCs in the culture supernatants of THP-1 cells. TNF-alpha and OSM, only when applied together, increased ALP activities and in vitro calcification in HVSMCs in the presence of IFN-gamma and 1,25(OH)2D3. These results suggest that macrophages may contribute to the development of vascular calcification through producing various inflammatory mediators, especially TNF-alpha and OSM.

  12. Evaluation of a chemiluminescence method for measuring alkaline phosphatase activity in whole milk of multiple species and bovine dairy drinks: interlaboratory study.

    Salter, Robert S; Fitchen, John


    Alkaline phosphatase (ALP) is a ubiquitous enzyme in milk with time-temperature destruction similar to that of certain pathogens destroyed in pasteurization. Measurement of ALP to indicate proper pasteurization is a common practice. Recently the public health level for ALP was decreased to 350 mU/L, a level below the sensitivity of older colorimetric ALP methods. This study was conducted within the structure of the International Dairy Federation and the International Organization for Standardization to evaluate the reproducibility of the chemiluminescence method (Charm PasLite) for ALP at 50, 100, 350, and 500 mU/L in whole milk of multiple species to meet new regulations in the United States and proposed regulations in the European Union (EU). Fifteen laboratories from 8 countries evaluated bovine, goat, sheep, and buffalo milk, bovine skim milk, 20% fat cream, and 2% fat chocolate milk. At ALP levels of 350 and 500 mU/L, the average relative standard deviation for repeatability (RSDr) was 7.5%, and the average relative standard deviation of reproducibility was (RSDR) 15%. For ALP at 100 and 50 mU/L, the average RSDr values were 10.5 and 12.6%, respectively, and the average RSDR values were 18 and 25%, respectively. The limit of detection was 20 mU/L. Results are comparable to those obtained with other enzymatic photo-activated system methods such as the fluorometric method. Results indicate that the method is suitable for measuring ALP in the milk of multiple species and in dairy drinks at U.S. and proposed EU levels.

  13. Display of E. coli Alkaline Phosphatase pIII or pVIII Fusions on Phagemid Surfaces Reveals Monovalent Decoration with Active Molecules

    Weichel, Michael; Jaussi, Rolf; Rhyner, Claudio; Crameri, Reto


    Active alkaline phosphatase of Escherichia coli (PhoA, EC was displayed via the leucine zipper element of the Jun-Fos heterodimer on the surface of filamentous phage and the kinetic parameters Km and kcat were determined. The phoA gene was cloned downstream of fos while jun was inserted upstream of pIII or pVIII, alternatively, in the pJuFo phagemid vector. Both fusion genes are regulated by independent lacZ promoters. PhoA displayed on the phagemid pIII surface exhibited a Km of 11.2 µM with 4-nitrophenyl phosphate as substrate, which is consistent with data published for soluble PhoA. Based on these data we calculated the decoration of pJuFo phagemid with PhoA using the minor and major coat proteins pIII and pVIII as fusion partners under variable inducing conditions. We found that, even if the promoters are fully induced at a concentration of 1000 µM IPTG, the phagemids display maximally one copy of PhoA-Fos-Jun-coat protein fusion, irrespective of whether the protein is presented via pIII or pVIII. However, since PhoA is displayed in a native-like fashion, as deduced from the kinetic parameters of the enzymatic reaction, the pJuFo technology provides a versatile tool for the functional screening of complex cDNA libraries displayed on the phagemids' surface. PMID:18949073

  14. Down regulation of a gene for cadherin, but not alkaline phosphatase, associated with Cry1Ab resistance in the sugarcane borer Diatraea saccharalis.

    Yunlong Yang

    Full Text Available The sugarcane borer, Diatraea saccharalis, is a major target pest of transgenic corn expressing Bacillus thuringiensis (Bt proteins (i.e., Cry1Ab in South America and the mid-southern region of the United States. Evolution of insecticide resistance in such target pests is a major threat to the durability of transgenic Bt crops. Understanding the pests' resistance mechanisms will facilitate development of effective strategies for delaying or countering resistance. Alterations in expression of cadherin- and alkaline phosphatase (ALP have been associated with Bt resistance in several species of pest insects. In this study, neither the activity nor gene regulation of ALP was associated with Cry1Ab resistance in D. saccharalis. Total ALP enzymatic activity was similar between Cry1Ab-susceptible (Cry1Ab-SS and -resistant (Cry1Ab-RR strains of D. saccharalis. In addition, expression levels of three ALP genes were also similar between Cry1Ab-SS and -RR, and cDNA sequences did not differ between susceptible and resistant larvae. In contrast, altered expression of a midgut cadherin (DsCAD1 was associated with the Cry1Ab resistance. Whereas cDNA sequences of DsCAD1 were identical between the two strains, the transcript abundance of DsCAD1 was significantly lower in Cry1Ab-RR. To verify the involvement of DsCAD1 in susceptibility to Cry1Ab, RNA interference (RNAi was employed to knock-down DsCAD1 expression in the susceptible larvae. Down-regulation of DsCAD1 expression by RNAi was functionally correlated with a decrease in Cry1Ab susceptibility. These results suggest that down-regulation of DsCAD1 is associated with resistance to Cry1Ab in D. saccharalis.

  15. Incorporation of alkaline phosphatase into layer-by-layer polyelectrolyte films on the surface of affi-gel heparin beads: physicochemical characterization and evaluation of the enzyme stability.

    Derbal, Lylia; Lesot, Hervé; Voegel, Jean Claude; Ball, Vincent


    The preparation of functionalized beads in the micrometer size range that can be used to probe the action of immobilized biomolecules on cell cultures during controlled periods of time is of fundamental importance in cell biology. However, the preparation and characterization of such particles is tedious because of their fast sedimentation. It is hence difficult to prepare such beads in a reproducible manner. This highlights the need to prepare an important batch of functionnalized particles and to store them under conditions where the loss of biological activity is minimized. The aim of this paper was to immobilize alkaline phosphatase (AP) as a model enzyme on the surface of Affi-gel heparin beads functionnalized by means of a layer-by-layer (LBL) film made of poly-l-glutamic (PGA) acid and poly-l-lysine (PLL). The enzyme has been adsorbed either on the top of the LBL film or embedded under five polyelectrolyte layers. When embedded, the enzyme was not released in buffer and retained more than 30% of its initial activity after 3 months of storage at 4 degrees C. However, when the enzyme was adsorbed on top of the LBL film, about 80% of the adsorbed enzyme was released in the buffer after a few days of storage. Longer storage did not lead to any further desorption and the remaining enzyme displayed the same evolution of its activity with time as the embedded enzyme. The time evolution of the enzyme activity on the beads is compared with that in solution alone and in the presence of PGA and PLL separately.

  16. Tetramisole and Levamisole Suppress Neuronal Activity Independently from Their Inhibitory Action on Tissue Non-specific Alkaline Phosphatase in Mouse Cortex.

    Nowak, Lionel G; Rosay, Benoît; Czégé, Dávid; Fonta, Caroline


    Tissue non-specific alkaline phosphatase (TNAP) may be involved in the synthesis of GABA and adenosine, which are the main inhibitory neurotransmitters in cortex. We explored this putative TNAP function through electrophysiological recording (local field potential ) in slices of mouse somatosensory cortex maintained in vitro. We used tetramisole, a well documented TNAP inhibitor, to block TNAP activity. We expected that inhibiting TNAP with tetramisole would lead to an increase of neuronal response amplitude, owing to a diminished availability of GABA and/or adenosine. Instead, we found that tetramisole reduced neuronal response amplitude in a dose-dependent manner. Tetramisole also decreased axonal conduction velocity. Levamisole had identical effects. Several control experiments demonstrated that these actions of tetramisole were independent from this compound acting on TNAP. In particular, tetramisole effects were not stereo-specific and they were not mimicked by another inhibitor of TNAP, MLS-0038949. The decrease of axonal conduction velocity and preliminary intracellular data suggest that tetramisole blocks voltage-dependent sodium channels. Our results imply that levamisole or tetramisole should not be used with the sole purpose of inhibiting TNAP in living excitable cells as it will also block all processes that are activity-dependent. Our data and a review of the literature indicate that tetramisole may have at least four different targets in the nervous system. We discuss these results with respect to the neurological side effects that were observed when levamisole and tetramisole were used for medical purposes, and that may recur nowadays due to the recent use of levamisole and tetramisole as cocaine adulterants.

  17. Ultrasensitive electroanalysis of low-level free microRNAs in blood by maximum signal amplification of catalytic silver deposition using alkaline phosphatase-incorporated gold nanoclusters.

    Si, Yanmei; Sun, Zongzhao; Zhang, Ning; Qi, Wei; Li, Shuying; Chen, Lijun; Wang, Hua


    An ultrasensitive sandwich-type analysis method has been initially developed for probing low-level free microRNAs (miRNAs) in blood by a maximal signal amplification protocol of catalytic silver deposition. Gold nanoclusters (AuNCs) were first synthesized and in-site incorporated into alkaline phosphatase (ALP) to form the ALP-AuNCs. Unexpectedly, the so incorporated AuNCs could dramatically enhance the catalysis activities of ALP-AuNCs versus native ALP. A sandwiched hybridization protocol was then proposed using ALP-AuNCs as the catalytic labels of the DNA detection probes for targeting miRNAs that were magnetically caught from blood samples by DNA capture probes, followed by the catalytic ligation of two DNA probes complementary to the targets. Herein, the ALP-AuNC labels could act as the bicatalysts separately in the ALP-catalyzed substrate dephosphorylation reaction and the AuNCs-accelerated silver deposition reaction. The signal amplification of ALP-AuNCs-catalyzed silver deposition was thereby maximized to be measured by the electrochemical outputs. The developed electroanalysis strategy could allow for the ultrasensitive detection of free miRNAs in blood with the detection limit as low as 21.5 aM, including the accurate identification of single-base mutant levels in miRNAs. Such a sandwich-type analysis method may circumvent the bottlenecks of the current detection techniques in probing short-chain miRNAs. It would be tailored as an ultrasensitive detection candidate for low-level free miRNAs in blood toward the diagnosis of cancer and the warning or monitoring of cancer metastasis in the clinical laboratory.

  18. Study of Topical Human Umbilical Cord Blood Serum in the Treatment of Alkaline Corneal Epithelial Wounds in Rabbit Model

    B Sharifi


    Full Text Available Introduction & Objective: One of the important functions of the cornea is to maintain normal vision by refracting light onto the lens and retina. This property is dependent in part on the ability of the corneal epithelium to undergo continuous renewal. Ocular surface failure which follows a variety of endogenous and exogenous precipitating factors, the most common being: chemical trauma, infection, alkaline burn, inflammation and hereditary conditions, lid or lash abnormalities, tear deficiency or reduced sensation. The core principal underpinning management strategy for ocular surface failure is establishing or promoting new growth of healthy conjunctiva and corneal epithelium. This process is mediated by many proteins that are inducers of corneal cell migration, proliferation, and differentiation. The current study was performed to investigate the efficacy of umbilical cord serum on alkaline corneal epithelial wound healing in the rabbit model. Materials & Methods: In this study conducted at Yasuj University of Medical Sciences in 2010, thirty two rabbits were randomly assigned into two equal groups. Central corneal alkali wound was formed in one eye of the rabbits by applying a 6-mm round filter paper, soaked in 1 N NaOH, for 60 seconds. Group one of animals received umbilical cord blood serum and group two received Sno*Tear in the eyes. The treatment was dosed 4 times a day with the eye drops, and epithelial wound closure was recorded using slit lamp. The data were analyzed to determine the rate of wound closure. Results: The mean wound radius closure rate was 0.77 mm/day (SD=0.013 for umbilical cord blood serum-treated eyes, 0.73 mm/day (SD=0.018 for artificial tear-treated eyes. Conclusion: This study shows that alkali-injured corneal epithelial wound heal faster when treated with umbilical cord blood serum than with artificial tear in rabbit model.

  19. The significance of early changes in serum alkaline-phosphatase levels following endocrine treatment in patients with advanced prostate cancer

    Aditya Pareek


    Conclusions: A significant proportion of patients with advanced carcinoma of the prostate have flare in SAP le-vels soon after initiating the treatment. We confirm that the flare activity in SAP levels is a negative prognostic indicator and it is possible to identify early treatment fail-ures by frequent measurements of SAP levels during first 6 weeks of commencing the therapy. The significance of SAP fare and androgen resistance is discussed.

  20. Serum total and bone alkaline phosphatase and tartrate-resistant acid phosphatase activities for the assessment of bone fracture healing in dogs

    C. Sousa


    Full Text Available O objetivo deste trabalho foi estudar o padrão de variação da atividade sérica da fosfatase alcalina total (tALP, da isoenzima óssea da fosfatase alcalina (BALP e da fosfatase ácida resistente ao tartarato (TRAP, assim como a variação da concentração dos minerais séricos durante o processo de cicatrização de fraturas ósseas no cão. A variação sérica destes marcadores do metabolismo ósseo foi avaliada em nove cães com fraturas diafisárias fechadas de ossos longos, submetidas a tratamento cirúrgico para osteosíntese. Durante o período pós-operatório, sete animais evoluíram no sentido de uma normal união óssea, sendo que dois deles desenvolveram um processo de não união óssea. Foram observados, relativamente à BALP, valores de actividade sérica mais elevados e com diferença estatística (P<0,05 no grupo de animais que evoluiu no sentido de uma normal união óssea, comparativamente ao grupo de animais que evoluiu no sentido do processo de não união. No grupo de animais que evoluiu para a completa união óssea foram, adicionalmente, observados valores diminuidos (P<0,05 da atividade sérica da TRAP, até ao dia 60 do período pós-operatório seguido de uma elevação estatisticamente significativa após este período. Em conclusão, os biomarcadores do metabolismo ósseo poderão vir a constituir um método auxiliar de diagnóstico na monitorização do processo de cicatrização de fracturas ósseas, possibilitando, a detecção precoce de complicações pós-operatórias.

  1. Development of a single-chain variable fragment-alkaline phosphatase fusion protein and a sensitive direct competitive chemiluminescent enzyme immunoassay for detection of ractopamine in pork

    Dong Jiexian; Li Zhenfeng; Lei Hongtao; Sun Yuanming [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Ducancel, Frederic [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Xu Zhenlin [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Boulain, Jean-Claude [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Yang Jinyi; Shen Yudong [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Wang Hong, E-mail: [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China)


    Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: Black-Right-Pointing-Pointer The scFv-AP fusion protein against ractopamine (RAC) was produced. Black-Right-Pointing-Pointer A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. Black-Right-Pointing-Pointer The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. Black-Right-Pointing-Pointer Recovery tests from pork samples were studied. Black-Right-Pointing-Pointer Good accuracy was obtained. - Abstract: A rapid, sensitive chemiluminescent enzyme immunoassay (CLEIA) for ractopamine (RAC) based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein was developed. The scFv gene was prepared by cloning the heavy- and light-chain variable region genes (V{sub H} and V{sub L}) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling V{sub H} and V{sub L} genes with a linker by means of splicing overlap extension polymerase chain reaction. The resulting scFv gene was inserted into the expression vector pLIP6/GN containing AP to produce the fusion protein in Escherichia coli strain BL21. The purified scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) protocol for detection of RAC. The average concentration required for 50% inhibition of binding and the limit of detection of the assay were 0.25 {+-} 0.03 and 0.02 {+-} 0.004 ng mL{sup -1}, respectively, and the linear response range extended from 0.05 to 1.45 ng mL{sup -1}. The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The results showed a good correlation between

  2. An in vitro study of alkaline phosphatase sensitivity to mixture of aflatoxin B1 and fumonisin B1 in the hepatopancreas of coastal lagoon wild and farmed shrimp Litopenaeus vannamei.

    Pérez-Acosta, Jesús A; Burgos-Hernandez, Armando; Velázquez-Contreras, Carlos A; Márquez-Ríos, Enrique; Torres-Arreola, Wilfrido; Arvizu-Flores, Aldo A; Ezquerra-Brauer, J Marina


    This study aimed to establish the combined effect of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on wild Litopenaeus vannamei hepatopancreas alkaline phosphatase (AP) activity compared with that of farmed shrimp. AP activity in hepatopancreas extract was confirmed by several specific inhibitor assays. AP activity of wild shrimp was higher than that of farmed shrimp (p vannamei was sensitive to the presence of both mycotoxins; however, AP is more sensitive to the combination of AFB1 + FB1 suggesting a possible synergistic or potentiating inhibitory effect.

  3. 骨形态发生蛋白7诱导骨膜细胞碱性磷酸酶的表达*%Bone morphogenetic protein-7 induces the expression of alkaline phosphatase in periosteal cells

    廖家成; 贝抗胜; 连银川


      背景:关于骨形态发生蛋白7作为刺激因子诱导细胞成骨的报道目前较少见。目的:观察骨膜细胞经骨形态发生蛋白7诱导后碱性磷酸酶的表达。  方法:取材于成人胫骨骨膜,常规细胞培养法行骨膜细胞体外培养,分为实验组和对照组,分别加入骨形态发生蛋白7加成骨细胞培养辅助剂和单纯成骨细胞培养辅助剂,相差显微镜观察骨膜细胞形态特征及超微结构。每组分别在第7,14,21天设3个时间点,每个时间点设3个样本,采用碱性磷酸酶试剂盒法检测成骨细胞特异性标志物碱性磷酸酶表达情况。  结果与结论:骨膜细胞经分组培养后,第7天时,实验组和对照组骨膜细胞均有明显增殖,碱性磷酸酶的可被检测出,但量不多,细胞外形为梭形,实验组比对照组检测的碱性磷酸酶数量稍多;第14天时,实验组及对照组骨膜细胞均显著增殖,细胞外形由梭形变为宽梭形,实验组比对照组检测的碱性磷酸酶数量明显增多。第21天时,实验组及对照组骨膜细胞均增殖,其中实验组细胞增殖明显,细胞外形为宽梭形,实验组比对照组检测的碱性磷酸酶数量显著增多。经过统计学分析由骨形态发生蛋白7诱导的骨膜细胞的成骨标志物碱性磷酸酶阳性率明显高于对照组(P OBJECTIVE:To study the expression of alkaline phosphatase of periosteal cel s after induced by bone morphogenetic protein-7 in vitro. METHODS:Periosteal cel s were obtained from adult tibial periosteum, and then the periosteal cel s were cultured by routine method in vitro. The cel s were divided into experimental group and control group, and then cultured with bone morphogenetic protein-7 plus osteoblast culture adjuvants and simple osteoblast culture adjuvants, respectively. The phase contrast microscope was used to observe the morphology and ultrastructure of periosteal

  4. Altered alkaline phosphatase activity in obese Zucker rats liver respect to lean Zucker and Wistar rats discussed in terms of all putative roles ascribed to the enzyme

    V. Bertone


    Full Text Available Biliary complications often lead to acute and chronic liver injury after orthotopic liver transplantation (OLT. Bile composition and secretion depend on the integrated action of all the components of the biliary tree, starting from hepatocytes. Fatty livers are often discarded as grafts for OLT, since they are extremely vulnerable to conventional cold storage (CS. However, the insufficiency of donors has stimulated research to improve the usage of such marginal organs as well as grafts. Our group has recently developed a machine perfusion system at subnormothermic temperature (20°C; MP20 that allows a marked improvement in preservation of fatty and even of normal rat livers as compared with CS. We sought to evaluate the response of the biliary tree of fatty liver to MP20, and a suitable marker was essential to this purpose. Alkaline phosphatase (AlkP, EC, frequently used as marker of membrane transport in hepatocytes and bile ducts, was our first choice. Since no histochemical data were available on AlkP distribution and activity in fatty liver, we have first settled to investigate AlkP activity in the steatotic liver of fatty Zucker rats (fa/fa, using as controls lean Zucker (fa/+ and normal Wistar rats. The AlkP reaction in Wistar rats was in accordance with the existing data and, in particular, was present in bile canaliculi of hepatocytes in the periportal region and midzone, in the canals of Hering and in small bile ducts but not in large bile ducts. In lean ZR liver the AlkP reaction in Hering canals and small bile ducts was similar to Wistar rat liver but hepatocytes had lower canalicular activity and besides presented moderate basolateral reaction. The difference between lean Zucker and Wistar rats, both phenotypically normal animals, could be related to the fact that lean Zucker rats are genotypically heterozygous for a recessive mutated allele. In fatty liver, the activity in ductules and small bile ducts was unchanged, but

  5. Comparison of alkaline phosphatase activity of MC3T3-E1 cells cultured on different Ti surfaces: modified sandblasted with large grit and acid-etched (MSLA), laser-treated, and laser and acid-treated Ti surfaces

    Li, Lin-Jie; Kim, So-Nam


    PURPOSE In this study, the aim of this study was to evaluate the effect of implant surface treatment on cell differentiation of osteoblast cells. For this purpose, three surfaces were compared: (1) a modified SLA (MSLA: sand-blasted with large grit, acid-etched, and immersed in 0.9% NaCl), (2) a laser treatment (LT: laser treatment) titanium surface and (3) a laser and acid-treated (LAT: laser treatment, acid-etched) titanium surface. MATERIALS AND METHODS The MSLA surfaces were considered as the control group, and LT and LAT surfaces as test groups. Alkaline phosphatase expression (ALP) was used to quantify osteoblastic differentiation of MC3T3-E1 cell. Surface roughness was evaluated by a contact profilometer (URFPAK-SV; Mitutoyo, Kawasaki, Japan) and characterized by two parameters: mean roughness (Ra) and maximum peak-to-valley height (Rt). RESULTS Scanning electron microscope revealed that MSLA (control group) surface was not as rough as LT, LAT surface (test groups). Alkaline phosphatase expression, the measure of osteoblastic differentiation, and total ALP expression by surface-adherent cells were found to be highest at 21 days for all three surfaces tested (P.05). CONCLUSION This study suggested that MSLA and LAT surfaces exhibited more favorable environment for osteoblast differentiation when compared with LT surface, the results that are important for implant surface modification studies. PMID:27350860

  6. Effects of Different Fertilizer on Soil Urease and Alkaline Phosphatase Activity%不同肥料对土壤脲酶和碱性磷酸酶活性的影响

    周俊国; 杨鹏鸣


    Ureas and alkaline phosphatase activity of soil in different fertilizer was studied in this experiment. The results shewed urea, chicken manure, straw increased soil urease activity by 15. 38 % , 55.94 % , 27.97 % and soil alkaline phosphatase activity by 2.31 % , 16.20 % , 10.42 % respectively compared with the control in the Ⅰ fertilization levrl. Three fertiliters byd soil urease activity by 24. 48 % ,67.83 % ,46.15 % and alkaline phosphatase activity by 9.72% , 28.01 % , 37.04 % reapectively in the Ⅱ fertilisation level. Urea, chicken manure, straw increased soil urease activity by 37.76 % , 86. 71 % , 62.24 % , and soil alkaline phosphatase activity by 28.47 % , 65.74 % , 58.10 % respectively in the Ⅲ fertilization level. The affect order for soil urease aclivity by 3 fertilizers in different fertilizer levels was chicken manure, straw,urea.The affecting order for soil alkaline phosphstase activity was chicken manure, stravt, urea in Ⅱ fertilization level. The affecting order for soil alkaline phoaphatase activity was straw, chicken manure, urea in Ⅰ and Ⅲ fertilization level.%本实验对不同施肥条件下土壤脲酶活性、碱性磷酸酶活性进行了系统研究.结果表明,和对照相比,尿素、鸡粪、秸秆在Ⅰ施肥水平上,使土壤脲(酶)活性分别增加15.38%、55.94%、27.97%,土壤碱性磷酸酶活性分别增加2.31%、16.20%、10.42%;在Ⅱ施肥水平上使土壤脲酶活性分别增加24.48%,67.83%,46.15%,使土壤碱性磷酸酶活性分别增加9.72%、28.01%和37.04%.在Ⅲ施肥水平上使土壤脲酶分别增加37.76%、86.71%、62.24%,土壤碱性磷酸酶活性分别增加28.47%、65.74%和58.10%.3种不同施肥水平对土壤腺酶活性影响顺序依次是鸡粪、秸秆、尿素.在Ⅱ施肥水平上,对土壤碱性磷酸酶活性的影响大小依次是鸡粪、秸秆、尿素.在Ⅰ、Ⅲ施肥水平上是秸秆、鸡粪、尿素.

  7. Analysis of detection results of bone alkaline phosphatase in 2 184 children in Baoji area%宝鸡地区2184例儿童骨碱性磷酸酶结果的分析

    乔正梅; 葛君琍; 赵秋剑; 张利强


    目的:评价骨碱性磷酸酶对儿童佝偻病早期诊断及疗效评价的临床价值,通过骨碱性磷酸酶的检测了解该地区儿童佝偻病的发病率。方法选择2013年1~12月来宝鸡市中心医院体检的2184例儿童,按年龄分成婴儿、幼儿、学龄前、学龄期4组,采集指血,采用北京中生金域诊断技术有限公司提供的骨源性碱性磷酸酶试剂盒检测末梢血中骨碱性磷酸酶水平。结果2184例受检儿童中,临床佝偻病(NBAP>250 U/L)的检出率为10.1%,且年龄越小检出率越高;亚临床佝偻病(NBAP 200~250 U/L)的检出率为65.5%,说明宝鸡地区大部分儿童处于亚健康状态;春冬季检出率高于夏秋季;NBAP 检出率无性别差异。结论儿童为生长发育较快时期,易引起维生素 D 缺乏,儿童骨碱性磷酸酶活性检测对维生素 D 缺乏引起的佝偻病的早期诊断有重要的临床价值。预防应从围产期开始,及时检测儿童骨碱性磷酸酶水平,以便早诊断、早治疗。%Objective To evaluate the clinical value of bone alkaline phosphatase in early diagnosis and treatment effect of chil-dren′rickets and to understand the incidence rate of rickets in children in the local region by the detection of neonatal bone alkaline phosphatase(NBAP).Methods 2 184 children with physical examination in our hospital from January to December 2013 were se-lected and divided into the infants,young children,preschool and school-age children groups according to age.The finger blood was collected for detecting the bone alkaline phosphatase in the peripheral blood by the bone alkaline phosphatase reagent kit provided by Beijing Jinyu diagnosis technology Co.,Ltd.Results Among detected 2 184 children,the detection rate of clinical rickets(NBAP>250 U/L)was 10.1 %,moreover the smaller the age,the higher the detection rate;the detection rate of subclinical rickets(NBAP 200~250 U/L)was 65

  8. ZZ亲和肽-碱性磷酸酶融合蛋白的制备与初步应用%Preparation of fusion protein between ZZ affinity peptide and alkaline phosphatase and its primary application

    鲍如梦; 赵爽佳; 薛敏; 杨洪鸣; 唐金宝


    目的:构建、表达ZZ亲和肽与碱性磷酸酶( AP)融合蛋白,并研究其生物学活性。方法:将ZZ亲和肽基因与AP基因重组,构建原核表达载体pEZZ-AP并利用大肠杆菌表达,金属离子亲和层析纯化目的蛋白,Western blot鉴定ZZ-AP生物学活性并初步应用于免疫细胞化学。结果:所构建重组质粒pEZZ-AP 经E.coli DH5α表达,SDS-PAGE结果显示目的蛋白相对分子量为62 kD,与理论值相近。 Western blot结果表明ZZ-AP融合蛋白既具有兔IgG抗体结合活性,又具有碱性磷酸酶活性,在细胞免疫组化应用中与AP标记的羊抗兔二抗显色模式与效果相似。结论:ZZ-AP融合蛋白构建成功,该融合蛋白具有兔IgG 抗体结合活性和碱性磷酸酶活性,细胞免疫组化应用结果表明该融合蛋白在免疫分析中具有潜在的应用价值。%Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic

  9. Pyridoxine-responsive seizures as the first symptom of infantile hypophosphatasia caused by two novel missense mutations (c.677T>C, p.M226T; c.1112C>T, p.T371I) of the tissue-nonspecific alkaline phosphatase gene.

    Baumgartner-Sigl, Sara; Haberlandt, Edda; Mumm, Steven; Scholl-Bürgi, Sabine; Sergi, Consolato; Ryan, Lawrence; Ericson, Karen L; Whyte, Michael P; Högler, Wolfgang


    Pyridoxine-responsive seizures (PRS) and the role of pyridoxine (PN, vitamin B(6)) in hypophosphatasia (HPP) are incompletely understood. Typically, PRS and HPP are rare, independent, metabolic disorders. In PRS, seizures resist standard anticonvulsants apart from PN, yet have a good prognosis. In HPP, inactivation of the tissue nonspecific isoenzyme of alkaline phosphatase (TNSALP) impairs skeletal mineralization and causes rickets in infants that can be fatal. Here, we report a 7-month-old girl, newly diagnosed with infantile HPP, who presented as a neonate with PRS but without bony abnormalities. Analysis of biogenic amines in cerebrospinal fluid (CSF) suggested brain pyridoxal 5'-phosphate (PLP) deficiency, although PLP in CSF was not decreased. She had normal cognitive milestones but failure to thrive and rickets. Nearly undetectable serum ALP activity, elevated plasma PLP and urinary phosphoethanolamine (PEA) and inorganic pyrophosphate (PPi) levels, hypercalcemia, hypercalciuria and nephrocalcinosis were consistent with infantile HPP. Only prednisolone reduced serum calcium levels. Despite improved growth and weight gain, she developed rib fractures and died from respiratory failure at age 9 months. Sequence analysis of the TNSALP gene revealed novel missense mutations in exon 7 (c.677T>C, p.M226T) and exon 10 (c.1112C>T, p.T371I). Our patient demonstrated that PRS in neonates may not necessarily be "idiopathic"; instead, such seizures can be caused by severe HPP that becomes clinically apparent later in infancy. The pathophysiology of PRS in HPP differs from the three other genetic defects known to cause PRS, but all may lead to brain PLP deficiency reducing seizure thresholds. All reported HPP patients with neonatal seizures died within 18 months of birth, suggesting that PRS is an indicator of HPP severity and lethal prognosis. We recommend that assessment of any neonate with PRS should include measurement of serum ALP activity.

  10. Age-related changes of serum tartrate-resistant acid phosphatase 5b and the relationship with bone mineral density in Chinese women

    Yue-juan QIN; Zhen-lin ZHANG; Hao ZHANG; Wei-wei HU; Yu-juan LIU; Yun-qiu HU; Miao LI; Jie-mei GU; Jin-wei HE


    Aim: Ostcoclastic activity is mainly assessed by measurement of urinary markers (eg C-terminal cross-linked telopeptides of type I collagen, N-terminal cross-linked telopeptides of type I collagen etc), the levels of which could often be affected by renal clearance. Recently, serum tartrate-resistant acid phosphatase 5b (TRACP5b) has been used as an alternative serum marker to evaluate osteoclastic activity. We investigated the age-related changes of TRACP5b level and its association with bone mineral density (BMD) in Chinese women. Methods: Seven-hundred and twenty-two Chinese mainland women aged 20-79 years were recruited in the study. Serum TRACP5b level was measured using immunoassay to evaluate the state of bone resorption. Bone mineral density (BMD) (g/cm2) at lumbar spine 1-4 and proximal femur were measured by duel-energy X-ray absorptiometry. Results: The serum TRACP5b level reached a bottom value in premenopausal women aged 30-39, gradually increased in women aged 40-49, rapidly rose in women aged 50-59, and culminated with a maximum value in women aged 60-69 before a slow drop in women aged 70-79. The average level of TRACPSb was significantly higher in postmenopausal women [(3.29±1.07) U/L] than in premenopausal women ([1.70±0.59] U/L). The levels of TRACP5b were inversely correlated with BMD at all measured sites (P<0.001). Furthermore, the level of TRACP5b was obviously higher in women with osteoporosis and osteopenia than those with normal bone mass (P<0.001). Conclusion: We have established the reference values of serum TRACPSb in Chinese mainland women, and found that postmenopausal women had higher TRACP5b concentration than younger women. The results showed that serum TRACPSb was a sensitive and useful parameter for the evaluation of age-related changes of bone absorption.

  11. The effects of fasting and refeeding on serum parathormone and calcitonin concentrations in young and old male rats.

    Kmieć, Z; Myśliwski, A; Wyrzykowska, M; Hoppe, A


    Although fasting and refeeding reveal the existence of age-related changes in carbohydrate and lipid metabolism, the effects of aging on mineral metabolism in refed animals are unknown. We therefore investigated hormonal regulation of calcium metabolism in young (4 months) and old (26 months) male rats fasted for 48 hours and then refed for 4 or 24 hours. Serum concentrations of total and ionized calcium and parathormone were similar in control young and old rats. Serum calcitonin level was higher, and the concentrations of albumin and inorganic phosphate and alkaline phosphatase activity were lower in fed old rats. In young fasted rats, the serum ionized and total calcium was decreased, and phosphate concentration was increased. In old rats, fasting resulted in the increase of serum parathormone level. Fasting reduced serum alkaline phosphatase activity to a similar extent in both age groups. In young rats, refeeding for 24h normalized serum calcium and phosphate levels and alkaline phosphatase activity, and decreased serum concentrations of PTH and calcitonin. In old refed rats, serum calcitonin concentration was raised by 77% compared to fed or fasted animals, whereas parathormone levels were normalized. Our results indicate that old fasted or refed rats maintain normal serum calcium concentration in a different way than young animals, possibly through the increase in serum levels of parathormone and/or calcitonin. Thus, dietary manipulations such as fasting and refeeding constitute an interesting model for the investigation of the effects of aging on the hormonal regulation of serum calcium level.

  12. A Chronoamperometric Screen Printed Carbon Biosensor Based on Alkaline Phosphatase Inhibition for W(VI Determination in Water, Using 2-Phospho-l-Ascorbic Acid Trisodium Salt as a Substrate

    Ana Lorena Alvarado-Gámez


    Full Text Available This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3, a repeatability of 9.4% (n = 3 and a detection limit of 0.29 ± 0.01 µM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case and a dynamic range from 0.6 to 30 µM. This study was performed by means of a Lineweaver–Burk plot, showing a mixed kinetic inhibition.

  13. A chronoamperometric screen printed carbon biosensor based on alkaline phosphatase inhibition for W(IV) determination in water, using 2-phospho-L-ascorbic acid trisodium salt as a substrate.

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia


    This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3), a repeatability of 9.4% (n = 3) and a detection limit of 0.29 ± 0.01 µM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case) and a dynamic range from 0.6 to 30 µM. This study was performed by means of a Lineweaver-Burk plot, showing a mixed kinetic inhibition.

  14. Stimulus Response of Au-NPs@GMP-Tb Core-Shell Nanoparticles: Toward Colorimetric and Fluorescent Dual-Mode Sensing of Alkaline Phosphatase Activity in Algal Blooms of a Freshwater Lake.

    Zhang, Xiaolei; Deng, Jingjing; Xue, Yumeng; Shi, Guoyue; Zhou, Tianshu


    In this study, we demonstrate a colorimetric and fluorescent dual-mode method for alkaline phosphatase activity (APA) sensing in freshwater lake with stimuli-responsive gold nanoparticles@terbium-guanosine monophosphate (Au-NPs@GMP-Tb) core-shell nanoparticles. Initially, the core-shell nanoparticles were fabricated based on Au-NPs decorated with a fluorescent GMP-Tb shell. Upon being excited at 290 nm, the as-formed Au-NPs@GMP-Tb core-shell nanoparticles emit green fluorescence, and the decorated GMP-Tb shell causes the aggregation of Au-NPs. However, the addition of ALP destroys GMP-Tb shell, resulting in the release of Au-NPs from the shell into the solvent. As a consequence, the aggregated Au-NPs solubilizes with the changes in the UV-vis spectrum of the dispersion, and in the meantime, the fluorescence of GMP-Tb shell turns off, which constitutes a new mechanism for colorimetric and fluorescent dual-mode sensing of APA. With the method developed here, we could monitor the dynamic change of APA during an algal bloom of a freshwater lake, both by the naked eye and further confirmed by fluorometric determination. This study not only offers a new method for on-site visible detection of APA but also provides a strategy for dual-mode sensing mechanisms by the rational design of the excellent optical properties of Au-NPs and the adaptive inclusion properties of the luminescent infinite coordination polymers.

  15. A Chronoamperometric Screen Printed Carbon Biosensor Based on Alkaline Phosphatase Inhibition for W(VI) Determination in Water, Using 2-Phospho-l-Ascorbic Acid Trisodium Salt as a Substrate

    Alvarado-Gámez, Ana Lorena; Alonso-Lomillo, María Asunción; Domínguez-Renedo, Olga; Arcos-Martínez, María Julia


    This paper presents a chronoamperometric method to determine tungsten in water using screen-printed carbon electrodes modified with gold nanoparticles and cross linked alkaline phosphatase immobilized in the working electrode. Enzymatic activity over 2-phospho-l-ascorbic acid trisodium salt, used as substrate, was affected by tungsten ions, which resulted in a decrease of chronoamperometric current, when a potential of 200 mV was applied on 10 mM of substrate in a Tris HCl buffer pH 8.00 and 0.36 M of KCl. Calibration curves for the electrochemical method validation, give a reproducibility of 5.2% (n = 3), a repeatability of 9.4% (n = 3) and a detection limit of 0.29 ± 0.01 μM. Enriched tap water, purified laboratory water and bottled drinking water, with a certified tungsten reference solution traceable to NIST, gave a recovery of 97.1%, 99.1% and 99.1% respectively (n = 4 in each case) and a dynamic range from 0.6 to 30 μM. This study was performed by means of a Lineweaver–Burk plot, showing a mixed kinetic inhibition. PMID:25621602

  16. Serum sialic acid and CEA concentrations in human breast cancer.

    Hogan-Ryan, A; Fennelly, J J; Jones, M; Cantwell, B; Duffy, M J


    The concentration of bound sialic acid in the sera of 56 normal subjects and 65 subjects with breast cancer was measured, in order to determine (1) whether serum sialic acid concentrations are raised in breast cancer and (2) whether the concentration of sialic acid in serum reflects tumour stage. The amount of sialic acid in serum was compared to serum carcinoembryonic antigen (CEA) values. Urinary hydroxyproline and serum alkaline phosphatase concentrations were used as indicators of bone and liver involvement. Erythrocyte sedimentation rate (ESR) was also measured. Significantly elevated serum sialic acid concentrations were found in breast cancer, and showed correlation with tumour stage. Serum sialic acid values did not correlate with CEA values. The results suggest that measurement of serum sialic acid concentrations may be of adjunctive value in assessing tumour stage.

  17. Detection and analysis on bone alkaline phosphatase activity of 2540 infants%2540例婴幼儿骨源性碱性磷酸酶活性检测分析



    目的:调查长兴地区0~5岁婴幼儿骨健康状况.方法:使用骨源性碱性磷酸酶(BAP)试剂盒对2009~2010年来长兴妇幼保健院门诊就诊的2 540例0~5岁婴幼儿进行检测.结果:在2 540例0~5岁婴幼儿中,BAP活性正常的1 215例,占总人数的47.83%;BAP活性处于预防水平的1 196例,占总人数的47.09%;BAP活性处于治疗水平的129例,占总人数的5.08%.结论:长兴地区约50.00%的婴幼儿骨骼处于亚健康状态,易患佝偻病,需及早预防.%Objective: To investigate the bone health status of 0 - 5 - year - old infants in Changxing county. Methods: Bone alkaline phosphatase ( BAP) activity of 2 540 infants aged 0-5 years who went to outpatient department of the hospital from 2009 to 2010 was detected by BAP kit. Results; Among2 540 infante aged 0 -5 years old, I 215 infants were found with normal BAP activity, accounting for 47. 83% ; BAP activity of 1 196 infants was at prevention level, accounting for 47.09% ; BAP activity of 129 infants was at treatment level, accounting for 5.08%. Conclusion: The bones of about 50.00% of the infants in Changxing county were subhealthy, the infants are susceptible to rickets, and prevention should be earned out early.

  18. Maternal antibiotic-induced early changes in microbial colonization selectively modulate colonic permeability and inducible heat shock proteins, and digesta concentrations of alkaline phosphatase and TLR-stimulants in swine offspring.

    Marie-Edith Arnal

    Full Text Available Elevated intake of high energy diets is a risk factor for the development of metabolic diseases and obesity. High fat diets cause alterations in colonic microbiota composition and increase gut permeability to bacterial lipopolysaccharide, and subsequent low-grade chronic inflammation in mice. Chronic inflammatory bowel diseases are increasing worldwide and may involve alterations in microbiota-host dialog. Metabolic disorders appearing in later life are also suspected to reflect changes in early programming. However, how the latter affects the colon remains poorly studied. Here, we hypothesized that various components of colonic physiology, including permeability, ion exchange and protective inducible heat shock proteins (HSP are influenced in the short- and long-terms by early disturbances in microbial colonization. The hypothesis was tested in a swine model. Offspring were born to control mothers (n = 12 or mothers treated with the antibiotic (ATB amoxicillin around parturition (n = 11. Offspring were slaughtered between 14 and 42 days of age to study short-term effects. For long-term effects, young adult offspring from the same litters consumed a normal or a palm oil-enriched diet for 4 weeks between 140 and 169 days of age. ATB treatment transiently modified maternal fecal microbiota although the minor differences observed for offspring colonic microbiota were nonsignificant. In the short-term, consistently higher HSP27 and HSP70 levels and transiently increased horseradish peroxidase permeability in ATB offspring colon were observed. Importantly, long-term consequences included reduced colonic horseradish peroxidase permeability, and increased colonic digesta alkaline phosphatase (AP and TLR2- and TLR4-stimulant concentrations in rectal digesta in adult ATB offspring. Inducible HSP27 and HSP70 did not change. Interactions between early ATB treatment and later diet were noted for paracellular permeability and concentrations of colonic

  19. QM/MM analysis suggests that Alkaline Phosphatase (AP) and Nucleotide pyrophosphatase/phosphodiesterase slightly tighten the transition state for phosphate diester hydrolysis relative to solution: implication for catalytic promiscuity in the AP superfamily

    Hou, Guanhua


    Several members of the Alkaline Phosphatase (AP) superfamily exhibit a high level of catalytic proficiency and promiscuity in structurally similar active sites. A thorough characterization of the nature of transition state for different substrates in these enzymes is crucial for understanding the molecular mechanisms that govern those remarkable catalytic properties. In this work, we study the hydrolysis of a phosphate diester, MpNPP−, in solution, two experimentally well-characterized variants of AP (R166S AP, R166S/E322Y AP) and wild type Nucleotide pyrophosphatase/phosphodiesterase (NPP) by QM/MM calculations in which the QM method is an approximate density functional theory previously parameterized for phosphate hydrolysis (SCC-DFTBPR). The general agreements found between these calculations and available experimental data for both solution and enzymes support the use of SCC-DFTBPR/MM for a semi-quantitative analysis of the catalytic mechanism and nature of transition state in AP and NPP. Although phosphate diesters are cognate substrates for NPP but promiscuous substrates for AP, the calculations suggest that their hydrolysis reactions catalyzed by AP and NPP feature similar synchronous transition states that are slightly tighter in nature compared to that in solution, due in part to the geometry of the bimetallic zinc motif. Therefore, this study provides the first direct computational support to the hypothesis that enzymes in the AP superfamily catalyze cognate and promiscuous substrates via similar transition states to those in solution. Our calculations do not support the finding of recent QM/MM studies by López-Canut and coworkers, who suggested that the same diester substrate goes through a much looser transition state in NPP/AP than in solution, a result likely biased by the large structural distortion of the bimetallic zinc site in their simulations. Finally, our calculations for different phosphate diester orientations and phosphorothioate diesters

  20. Field-Evolved Mode 1 Resistance of the Fall Armyworm to Transgenic Cry1Fa-Expressing Corn Associated with Reduced Cry1Fa Toxin Binding and Midgut Alkaline Phosphatase Expression.

    Jakka, Siva R K; Gong, Liang; Hasler, James; Banerjee, Rahul; Sheets, Joel J; Narva, Kenneth; Blanco, Carlos A; Jurat-Fuentes, Juan L


    Insecticidal protein genes from the bacterium Bacillus thuringiensis (Bt) are expressed by transgenic Bt crops (Bt crops) for effective and environmentally safe pest control. The development of resistance to these insecticidal proteins is considered the most serious threat to the sustainability of Bt crops. Resistance in fall armyworm (Spodoptera frugiperda) populations from Puerto Rico to transgenic corn producing the Cry1Fa insecticidal protein resulted, for the first time in the United States, in practical resistance, and Bt corn was withdrawn from the local market. In this study, we used a field-collected Cry1Fa corn-resistant strain (456) of S. frugiperda to identify the mechanism responsible for field-evolved resistance. Binding assays detected reduced Cry1Fa, Cry1Ab, and Cry1Ac but not Cry1Ca toxin binding to midgut brush border membrane vesicles (BBMV) from the larvae of strain 456 compared to that from the larvae of a susceptible (Ben) strain. This binding phenotype is descriptive of the mode 1 type of resistance to Bt toxins. A comparison of the transcript levels for putative Cry1 toxin receptor genes identified a significant downregulation (>90%) of a membrane-bound alkaline phosphatase (ALP), which translated to reduced ALP protein levels and a 75% reduction in ALP activity in BBMV from 456 compared to that of Ben larvae. We cloned and heterologously expressed this ALP from susceptible S. frugiperda larvae and demonstrated that it specifically binds with Cry1Fa toxin. This study provides a thorough mechanistic description of field-evolved resistance to a transgenic Bt crop and supports an association between resistance and reduced Cry1Fa toxin binding and levels of a putative Cry1Fa toxin receptor, ALP, in the midguts of S. frugiperda larvae.

  1. Crystallographic studies on shrimp alkaline phosphatase

    Backer, M.M.E. (Maaike Maria Eva)


    The earth has large cold areas, such as mountains, oceans and (ant)arctic regions, in which organisms have evolved to survive. This adaptation happens at a molecular level. The question is, how do proteins adjust such that they function at low temperatures? "Cold-active" or "cold-adapted" enzymes ha

  2. Crucial role of alkaline sphingomyelinase in sphingomyelin digestion: a study on enzyme knockout mice

    Zhang, Yao; Cheng, Yajun; Hansen, Gert H


    ) and KO mice were fed ³H-palmitic acid labeled SM together with milk SM by gavage. The lipids in intestinal content, intestinal tissues, serum, and liver were analyzed by TLC. In KO mice, nondigested ³H-SM in the intestinal content increased by 6-fold and the formation of ³H-ceramide decreased markedly....... The KO mice also showed significantly decreased radioactivity in liver and serum. Furthermore, alkaline phosphatase activity in the mucosa was reduced by 50% and histological comparison of two female littermates preliminarily suggested mucosal hypertrophy in KO mice. This study provides definite proof...

  3. [Effect of VAM fungi on phosphatase activity in maize rhizosphere].

    Song, Y; Li, X; Feng, G


    The effect of VAM fungi on phosphatase activity in maize rhizosphere was examined by pot culture experiment, in which, three-compartment-pots were used, the central compartment being separated from the outer two by a nylon net with 30 microns mesh. Plants were harvested 70 days after planting. Soil acid and alkaline phosphatase were measured at different distances from root surface. The results showed that VAM increased the activities of soil acid and alkaline phosphatase in the rhizosphere. It was found that different phosphorous sources had different effects on phosphatase activity.

  4. [Phosphatase activity in Amoeba proteus at pH 9.0].

    Sopina, V A


    In the free-living amoeba Amoeba proteus (strain B), after PAAG disk-electrophoresis of the homogenate supernatant, at using 1-naphthyl phosphate as a substrate and pH 9.0, three forms of phosphatase activity were revealed; they were arbitrarily called "fast", "intermediate", and "slow" phosphatases. The fast phosphatase has been established to be a fraction of lysosomal acid phosphatase that preserves some low activity at alkaline pH. The question as to which particular class the intermediate phosphatase belongs to has remained unanswered: it can be both acid phosphatase and protein tyrosine phosphatase (PTP). Based on data of inhibitor analysis, large substrate specificity, results of experiments with reactivation by Zn ions after inactivation with EDTA, other than in the fast and intermediate phosphatases localization in the amoeba cell, it is concluded that only slow phosphatase can be classified as alkaline phosphatase (EC

  5. Effect of phenylmercuric acetate injections on phosphatase activity in chickens resistant and susceptible to Leukosis

    Miller, V.L.; Bearse, G.E.; Csonka, E.


    The weighted means of liver and kidney alkaline phosphatase activity was greater in three strains of chickens classified as susceptible to limphoid leukosis than in five strains classified as resistant. On the same basis, four strains classified as susceptible to Marek's disease had more liver alkaline phosphatase activity than four strains classified as resistant. The weighted means of liver and kidney acid phosphatase activity were not different among the same strains of chickens classified similarly. Kidney alkaline phosphatase activity was the most generally inhibited by phenylmercuric acetate injections, followed by liver acid and alkaline phosphatase. Kidney acid phosphatase activity was enhanced by phenylmercuric acetate injections in three strains of chickens classified as resistant to both lymphoid leukosis and Marek's disease. Liver acid phosphatase activity was depressed in three strains classed as resistant to lymphoid leukosis.

  6. Value of detection of calcium in whole blood and bone alkaline phosphatase in the diagnosis of rickets%全血钙和骨碱性磷酸酶检测在佝偻病诊断中的应用



    目的 探讨全血钙和骨碱性磷酸酶(BALP)在儿童维生素D缺乏性佝偻病中的诊断价值.方法 对755例体检儿童采末梢血,用全血干化学法和免疫浓缩技术检测BALP,全血钙检测采用原子吸收光谱分析法.结果 755例体检儿童中BALP≤200 U/L 138例,200~250 U/L 386例,≥250U/L231例.血钙<1.55 mmol/L 79例,≥1.55mmol/L676例.BALP异常检出率为81.7%(617/755),血钙异常检出率为10.5%(79/755).血钙在各年龄组和男女性别间差异均无统计学意义.BALP在男女性别间差异无统计学意义,在不同年龄组间差异有统计学意义,1岁以下组BALP异常检出率最高,3~5岁组最低.大部分儿童处于维生素D轻度缺乏或缺乏状态,且随着年龄增长发病率有逐渐降低的趋势.结论 BALP的检测在维生素D缺乏性佝偻病诊断中较血钙测定敏感,更能反映人体钙营养水平.为更好地预防、控制和治疗佝偻病,婴幼儿应定期做相关项目的健康体检.%Objective To investigate the diagnostic value of calcium in whole blood and bone alkaline phosphatase (BALP) in vitamin D deficiency rickets in children. Methods Peripheral blood was collected from 755 children. Levels of BALP were detected based on dry chemical and immune concentrated technique, and calcium was detected by atomic absorption spectroscopy. Results BALP levels were showed ≤200 U/L in 138 children, 200-250 U/L in 386 children, and ≥ 250 U/L in 231 children. Calcium was found <1.55 mmo1/L in 79 children and ≥ 1.55 mmol/L in 676 children. The detection rate of abnormal BALP was 81.7% (617/755), and that of abnormal calcium was 0.5% (79/755). Calcium showed no statistically significant difference between different age groups and gender groups. The BALP showed no statistically significant difference between males and females, but significant differences between different age groups. The detection rate of abnormal BALP was highest in age group that smaller than one

  7. 环境因子对湖泊沉积物碱性磷酸酶活性的影响%Impact of Environmental Factors on Alkaline Phosphatase Activity of Sediment in Lakes

    张翠英; 徐德兰; 万蕾; 琚淑明; 高明侠


    Alkaline phosphatase activity(APA) of sediment in Yunlong Lake and Luoma Lake was determined, and effects of physical and chemical characteristics of water, seasonal dynamics and aquatic plants on APA of different lakes were analyzed. Results showed that APA was significantly positive correlation with pH value (r=0.220, P<0.05) and temperature (r=0.478, P<0.01) respectively, but DO had little effect on APA Seasonal change of APA in Yunlong Lake was obvious, with the highest APA of 330.27 mg/(kg·h) in summer and the lowest of 154.65 mg/(kg·h) in spring. pH value varied from 8.12 to 8.50, temperature varied from 3.3 t to 26.5 ℃, DO varied from 6.84 mg/L to 8.47 mg/L. Aquatic plants had great effect on APA of Luoma Lake, APA of grass area was greater than that of no grass area, and seasonal change of APA was obvious in grass area. In brief, physical and chemical characteristics of water, seasonal dynamics and aquatic plants have great effect on APA.%文章对云龙湖和骆马湖2种不同类型湖泊沉积物的碱性磷酸酶进行了测定,分析了水体理化特性、季节变化及水生植物对湖泊沉积物碱性磷酸酶活性的影响,结果表明:pH值、温度对沉积物碱性磷酸酶活性影响较大,呈显著正相关(r=0.220,P<0.05; r=0.478,P<0.01),而DO值的影响较小,相关不显著;随着季节的变化,云龙湖沉积物碱性磷酸酶活性变化较明显,其变化规律为夏季>秋季>冬季>春季,夏季最高,为330.27 mg/(kg·h),春季最低,为154.65 mg/(kg·h);随着季节的变化,水体pH、DO变化较小,分别为8.12~8.50、6.84~8.47 mg/L,温度变化较大,为3.3~26.5℃;水生植物对骆马湖沉积物碱性磷酸酶活性影响较大,有草区均大于无草区且有草区碱性磷酸酶活性随季节变化显著.因此,水体理化特性、季节变化及水生植物等环境因子对不同湖泊沉积物中碱性磷酸酶活性有重要影响.

  8. Estudio de la fosfatasa ácida y alcalina en suelos de la Región Pampeana Norte del área sojera argentina Study of acid and alkaline phosphatase in soils of the Pampean North Region from argentine soybean area

    Leticia Andrea Fernández


    ón de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po.Transformation of organic phosphorus (Po into soluble inorganic phosphorus (Pi is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB and cultivated fungi (CF was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5 CFU g-1 soil, while alkaline activity was 5.80 10(5 CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5 mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

  9. Chromogranin A as serum marker for gastroenteropancreatic neuroendocrine tumors: a single center experience and literature review.

    Christoph J Auernhammer; Christine Spitzweg; Burkhard Göke; Hoffmann, Johannes N.; Herrmann, Karin A.; Alexander Haug; Michael Vogeser; Axel Kuttner; Michael Lauseker; Svenja Nölting


    The aim of this study was to assess the clinical sensitivities of the tumor markers chromogranin A (CgA), urinary 5-hydroxyindoleacetic acid (5-HIAA) and alkaline phosphatase (AP) in neuroendocrine tumors (NETs) of the GastroEnteroPancreatic-(GEP-) system depending on tumor primary location and metastatic spread. In a retrospective single-center series, sensitivities were evaluated in serum samples from 110 patients with midgut (n = 62) and pancreatic (n = 48) NETs. CgA levels were analyzed b...

  10. 骨碱性磷酸酶检测技术诊治小儿佝偻病在浙江农村推广的适宜性评估%Evaluation of diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection in township-level health organizations of Zhejiang rural areas

    朱雯; 朱炜; 杨再峰; 王红妹; 张扬; 沈毅; 李鲁; 姒健敏; 蒋健敏; 夏志俊; 蒋海瑛


    Objective: To evaluate the health technology of diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection in township-level health organizations of Zhejiang rural areas. Methods: The health technology was evaluated by the assessment model for selection and assessment of rural appropriate health technology in Zhejiang province. Results: The weighted score of first-level indicators were 8.05, 8.06, 6.32, 18.13, 18.88, 4.84 and 4.36 respectively. The total score was 68.64. Maturity: good; need: fair; convenience: good; safety: good; validity: very good; economic attributes: fair; sociality and ethicality: good. Conclusion: Diagnosis of infantile vitamin D deficiency rickets by bone alkaline phosphatase detection is suitable for dissemination and application in township-level health organizations in Zhejiang rural areas.%目的:对浙江省农村乡级卫生机构在小儿佝偻病诊治中应用骨碱性磷酸酶检测技术进行系统全面的评估.方法:使用"浙江省农村卫生适宜技术筛选评估指标体系"进行评估.结果:7个一级指标的加权分分别为8.05、8.06、6.32、18.13、18.88、4.84、4.36,总分为68.64;该技术成熟性较好;需要性一般;简便性较好;安全性较好;有效性很好;经济性一般;社会和伦理适应性较好.结论:该技术适宜在浙江省乡级农村卫生机构推广应用.

  11. Passive immunity transfer and serum constituents of crossbred calves

    Thaís G. Rocha


    Full Text Available Passive immunity transfer (PIT evaluation is an essential tool for the maintenance of healthy calves during the first months of life. Since lactation number and breed have been proven to influence immunoglobulin levels in colostrum, the aim of this study was to evaluate PIT from primiparous and multiparous Canchim cows to their calves. Blood samples were collected from the calves before colostrum intake and 1, 2, 7, 15 and 30 days thereafter, while colostrum samples from the cows were taken immediately after parturition. Activities of gamma-glutamyl transferase (GGT, alkaline phosphatase (ALP, and concentrations of total protein, albumin, globulins, immunoglobulin A (IgA, immunoglobulin G (IgG, total and ionized calcium, inorganic phosphorus, magnesium, sodium and potassium were evaluated in calves' serum and activities of GGT and ALP and concentrations of total protein, IgA and IgG were assessed in cow's colostrum whey. Immunoglobulins concentrations were evaluated by electrophoresis in polyacrylamide gels. Serum biochemistry evaluations revealed an increase in gamma-glutamyl transferase and alkaline phosphatase activities and in total protein, globulins, immunoglobulin A and immunoglobulin G levels in calves' serum after colostrum intake. Only total protein and light chain immunoglobulin G levels in colostrum whey were affected by the cows' lactation number. Phosphorus and magnesium levels in blood serum increased after colostrum intake, while sodium and potassium levels oscillated in the experimental period. PIT was influenced by the cows' lactation number but was efficient in both groups.

  12. Hemograma, bioquímica sérica e histologia da biópsia hepática de bovinos após administração de polpa cítrica Hemogram, serum biochemistry and hepatic histologic features in cattle after administration of citrus pulp

    N.J.F. Oliveira


    Full Text Available Hemogram and serum biochemistry (aspartate aminotransferase, alkaline phosphatase and gamma glutamiltransferase, total protein, urea, creatinine, calcium and phosphorus were performed weekly in five crossbreed bovine after consumption of a diet containing citrus pulp pellets (40%, for 43 days. Percutaneous hepatic biopsy and histologic evaluation were performed in each animal before and after consumption of the citrus pulp diet. Hemogram, the enzymes aspartate aminotransferase and gamma glutamiltransferase, urea and creatinine had normal levels at the end of the experiment. No histologic lesions were observed in liver samples before or after citrus pulp consumption. However, there was an increase of serum phosphorus and reduction of serum calcium (p<0.05, without hypercalcemia, after consumption of citrus pulp diet. There was also an increase in serum alkaline phosphatase (p<0.05, probably induced by bone isoenzyme.

  13. Phosphatase activity in the rhizosphere and root of mycorrhizal teak seedlings with three levels of NPK fertilization



    Full Text Available To examine the phosphatase alkaline activity of VA mycorrhizal fungi in the rizhosphere and in root, teak seedlings inoculated spores of VA mycorrhizal fungi were grown in sterilized soils. Teak seedlings were fertilized with NPK fertilizer consisting three levels, i.e. 0; 0.0625; 0.125 g per seedling. Phosphatase alkaline in rizhosphere was measured in terms of pNP on soil dry weight basis, meanwhile alkaline phosphatase activity in roots were quantified in using method developed by Tisserant. The results showed that alkaline phosphatase activity increased on inoculated seedlings compare to with uninoculated. NPK fertilization of 0.0625 g per seedling and inoculation on teak seedlings showed alkaline phosphatase activity in range 90-201 EU, and in roots indicated in range 14-72%. Gigaspora sp inoculation on teak seedlings was showing the highest of alkaline phosphatase activity. Increasing phosphatase alkaline activity relevant to hyphae growth, and increasing of root infection decreased alkaline phosphatase activity. Arbuscular mycorrhizal inoculation increased seedling dry weight.

  14. Serum chemistry profiles for Lechwe waterbucks (Kobus leche): variations with age and sex.

    Váhala, J; Kase, F


    1. Over an 8-year period, 19 biochemical parameters have been determined at various ages in the blood serum of 92 clinically healthy Lechwe waterbucks (Kobus leche), 33 males and 59 females. 2. Significant differences have been noted with age. In neonates, the lowest values of total proteins, glucose, creatinine, urea, AST, ALT and iron have been noted; the highest ones have been seen for cholesterol, alkaline phosphatase, calcium and phosphorus. 3. With regard to sex, raised values of glucose, urea, alkaline phosphatase and ALT, and lowered values of cholesterol, have been noted in juvenile females compared with males of the same age. 4. In adult females, higher levels of urea and cholesterol and lower levels of glucose, triglycerides and natrium have been recorded compared with males. 5. With sex and age, no significant changes have been found in the levels of GGT, magnesium, chlorides and copper. 6. Our findings are discussed with those abstracted from the literature for related species.

  15. [ATPase and phosphatase activity of drone brood].

    Bodnarchuk, L I; Stakhman, O S


    Most researches on insect enzymes concern carbohydrate and nitrogenous exchange. Data on ATPase activity for larval material of drone brood are absent in the available literature. The drone brood is one of the least investigated apiproducts. Allowing for the important role of ATPase in the vital functions of the insect cells our work was aimed at the study of ATPase of the drone blood activity and that of alkaline and acid phosphatases. When studying liophylised preparations of the drone brood homogenate we have found out high activity of Mg2+, Na+, K+-, Ca2+- and Mg2+-ATPase and of alkaline and acid phosphatase, that is the possible explanation of the high-intensity power and plastic processes proceeding during growth and development of larvae.

  16. Detection of phosphatase activity in aquatic and terrestrial cyanobacterial strains

    Babić Olivera B.


    Full Text Available Cyanobacteria, as highly adaptable microorganisms, are characterized by an ability to survive in different environmental conditions, in which a significant role belongs to their enzymes. Phosphatases are enzymes produced by algae in relatively large quantities in response to a low orthophosphate concentration and their activity is significantly correlated with their primary production. The activity of these enzymes was investigated in 11 cyanobacterial strains in order to determine enzyme synthesis depending on taxonomic and ecological group of cyanobacteria. The study was conducted with 4 terrestrial cyanobacterial strains, which belong to Nostoc and Anabaena genera, and 7 filamentous water cyanobacteria of Nostoc, Oscillatoria, Phormidium and Microcystis genera. The obtained results showed that the activity of acid and alkaline phosphatases strongly depended on cyanobacterial strain and the environment from which the strain originated. Higher activity of alkaline phosphatases, ranging from 3.64 to 85.14 μmolpNP/s/dm3, was recorded in terrestrial strains compared to the studied water strains (1.11-5.96 μmolpNP/s/dm3. The activity of acid phosphatases was higher in most tested water strains (1.67-6.28 μmolpNP/s/dm3 compared to the activity of alkaline phosphatases (1.11-5.96 μmolpNP/s/dm3. Comparing enzyme activity of nitrogen fixing and non-nitrogen fixing cyanobacteria, it was found that most nitrogen fixing strains had a higher activity of alkaline phosphatases. The data obtained in this work indicate that activity of phosphatases is a strain specific property. The results further suggest that synthesis and activity of phosphatases depended on eco-physiological characteristics of the examined cyanobacterial strains. This can be of great importance for the further study of enzymes and mechanisms of their activity as a part of cyanobacterial survival strategy in environments with extreme conditions. [Projekat Ministarstva nauke Republike

  17. Spatial-temporal Variations of Alkaline Phosphatase Activity in the Kuilei Lake and Their Influencing Factors%傀儡湖水体中碱性磷酸酶活性的时空变化及其影响因素

    张成艳; 成小英; 王建军; 张路; 杜应旸


    Alkaline phosphatase activity (APA) plays an important role in the biogeochemical cycling of phosphorus.The spatial-temporal distribution of APA and related environmental variables were studied in the Kuilei Lake.The research found significant seasonal and regional variations of APA in the water body,with lower APAs in outlet waters and higher in the estuarine.APAs were relatively low in winter (January),ranging from 2.06 to 17.33 nmol mL-1 h-1,and high in summer (July),ranging from 5.78 to 13.52 nmol mL-1 h-1.Correlation analyses indicated that APAs were significantly related to particulate organic nitrogen,total nitrogen and NO2--N.Other nitrogen forms (NO3--N and NH4+-N) were also related,though not significantly.The results implied that the factors related to nitrogen are the primary factors affecting APAs.%碱性磷酸酶在磷的生物地球化学循环过程中发挥着重要作用.以傀儡湖为研究区域,对其水体中碱性磷酸酶活性(Alkaline phosphatase activity,APA)及其理化性质的时空分布进行分析.结果表明,傀儡湖水体APA呈时空异质性:河道较低,河口则易出现较高的APA;全湖的APA存在较强的季节性变化,冬季(1月)最低,变化范围为2.06-17.33nmol mL-1h-1;夏季(7月)最高,变化范围为5.78-13.52 nmol mL-1 h-1.APA与环境因子相关性分析显示,APA与颗粒态有机氮浓度关系最为密切,与总氮含量的相关性其次,随后显著相关的因子为NO2--N,此外是NO3--N和NH4+-N.这一结果表明与氮元素有关的环境因子可能是影响APA的主要因素.


    S. Selvapandian Kirubaharan


    Full Text Available INTRODUCTION Vitamin D metabolites participate in the regulation of calcium homoeostasis and bone metabolism. Severe and prolonged vitamin D deficiency causes osteomalacia. Subclinical vitamin D deficiency is common in the elderly and may lead to development of secondary hyperparathyroidism and bone loss, for which reason it has been implicated in the pathogenesis of senile osteoporosis. AIM To determine the relationships between serum 25 (OH D, bone mineral density, and serum calcium and phosphorus in 50 postmenopausal women. MATERIALS AND METHODS A population-based cross-sectional study with a total of 50 healthy women aged over 50 years with amenorrhoea for 12 months were included. None of the women were using hormone replacement therapy. Measurements of bone mineral density were performed by dual-energy X-ray absorptiometry (DXA. Analysis of serum levels of 25(OH D, calcium, phosphate and alkaline phosphatase were done. RESULTS There exists an inverse correlation between BMD and 25(OH D levels (r = - 0.127, p>0.05 and that between 25 (OH D and phosphate levels among postmenopausal women (r = -0.167, p >0.05, which is not significant. No significant difference exists in the serum levels of calcium, alkaline phosphatase between vitamin D insufficient & deficient persons (p >0.05. CONCLUSION Elevated levels of serum 25(OH D were associated with decreased bone mineral density and increased bone turnover, but it was insignificant.

  19. Serum chemistry of the collared peccary (Tayassu tajacu).

    Lochmiller, R L; Grant, W E


    Values of serum biochemistry were obtained for 33 adult (16 male and 17 female) and six juvenile (four male and two female) collared peccaries collected by trapping and drugging animals from southern Texas during the period July through September 1982. Only cholesterol and aspartate aminotransferase concentrations differed significantly with respect to sex. Juvenile peccaries had significantly lower concentrations of total protein and globulins, but had higher concentrations of glucose, triglycerides, alkaline phosphatase, alanine aminotransferase, and calcium, and a higher albumin/globulin ratio than did adults. Effects of method of capture on biochemical attributes of serum from five gunshot and nine trapped adult peccaries collected from a single herd during March 1983 also were examined. Trapped peccaries had significantly higher levels of serum urea nitrogen, urea nitrogen/creatinine ratio, glucose, and chloride concentrations. Levels of uric acid, calcium, and potassium were significantly lower among the trapped animals.

  20. Serum cross-linked n-telopeptides of type 1 collagen (NTx in patients with solid tumors

    Fernando Jablonka

    Full Text Available CONTEXT AND OBJECTIVE: Cross-linked N-telopeptides of type I collagen (NTx increase in concentration in situations in which bone resorption is increased, such as osteoporosis and bone metastasis (BM. We aimed to evaluate the serum concentrations of NTx in a sample of patients with several types of solid tumors. DESIGN AND SETTING: Cross-sectional analytical study with a control group in a tertiary public hospital. METHODS: We performed the quantitative enzyme-linked immunosorbent assay (ELISA on serum NTx levels in 19 subjects without a history of cancer and 62 patients with various solid tumors who had been referred for a bone scan. Three experienced analysts read all bone scans. RESULTS: The serum NTx levels in patients with cancer and BM, with cancer but without BM and without cancer were 46.77 ± 2.58, 32.85 ± 2.05 and 22.32 ± 2.90 respectively (P < 0.0001. We did not find any significant correlations of serum NTx with age, gender, history of bone pain, tumor type and bone alkaline phosphatase levels. We found a significant correlation between serum NTx and alkaline phosphatase levels (R² = 0.08; P = 0.022. CONCLUSIONS: Serum NTx levels are significantly higher in patients with solid tumors and bone metastases than they are in patients without bone metastases and in normal controls.

  1. Effects of Metal Ions and Organic Solvents on Alkaline Phosphatase from Rice-field Eel%金属离子·有机溶剂对黄鳝碱性磷酸酶的影响

    黄毅; 唐云明


    [Objective]The mechanism of alkaline phcephatase(ALP) was studied to promote rice-field eel aquaculture industry. [Method] The effects of effectors such as multiple metal ions and organic solvents on ALP in viscera of rice-field eel. [ Result] Na+ and K+ didn't generate big influences on en-zyme activity ;Mg2+ and Ca2+ could promote ALP while Li+,Cu2+ and Zn2+ could restrain ALP enzyme activity. Both KPO2-4 and WO3-4 generated by en-zyme catalyzing disodium phenyl phosphate possessed strong inhibitory effects on emzyme, and 9.5 mmol/L HPO2-4 would make enzyme activity decline by 13% while 9.5 mmol/L WO3-4 would make enzyme decline by 34%. The inhibition types of them were both competitive inhibition on enzyme activity. The organic solvents such as methanol,ethanol,ethylene glycol,isopropanol all generated influences on ALP and the order according to their inhibitory effects was isopropanol > ethanol > methanol > ethylene glycol. [Conclusion] The influences of various effectors on ALP activity of rice-field eel were studied from dy-namics perspective to provide theoretical basis for further clarifying ALP mechanism.

  2. Establishment and assessment of two methods for quantitative detection of serum duck hepatitis B virus DNA

    Ya-Xi Chen; Ai-Long Huang; Zhen-Yuan Qi; Shu-Hua Guo


    AIM: To establish and assess the methods for quantitative detection of serum duck hepatitis B virus (DHBV) DNA by quantitative membrane hybridization using DHBV DNA probe labeled directly with alkaline phosphatase and fluorescence quantitative PCR (qPCR).METHODS: Probes of DHBV DNA labeled directly with alkaline phosphatase and chemiluminescent substrate CDP-star were used in this assay. DHBV DNA was detected by autoradiography,and then scanned by DNA dot-blot. In addition, three primers derived from DHBV DNA S gene were designed. Semi-nested primer was labeled by AmpliSensor. Standard curve of the positive standards of DHBV DNA was established after asymmetric preamplification, semi-nested amplification and on-line detection. Results from 100 samples detected separately by alkaline phosphatase direct-labeled DHBV DNA probe with dot-blot hybridization and digoxigeninlabeled DHBV DNA probe hybridization. Seventy samples of duck serum were tested by fluorescent qPCR and digoxigeninlabeled DHBV DNA probe in dot-blot hybridization assay and the correlation of results was analysed.RESULTS: Sensitivity of alkaline phosphatase direct-labeled DHBV DNA probe was 10 pg. The coincidence was 100%compared with digoxigenin-labeled DHBV DNA probe assay.After 30 cycles, amplification products showed two bands of about 180 bp and 70 bp by 20 g/L agarose gel electrophoresis.Concentration of amplification products was in direct proportion to the initial concentration of positive standards. The detection index was in direct proportion to the quantity of amplification products accumulated in the current cycle.The initial concentration of positive standards was in inverse proportion to the number of cycles needed for enough quantities of amplification products. Correlation coefficientof the results was (0.97, P<0.01) between fluorescent qPCRand dot-blot hybridization.CONCLUSION: Alkaline phosphatase direct-labeled DHBV DNA probe in dot-blot hybridization and fluorescent qPCR can be

  3. Observation on the activities of alpha-L-fucosidase,alkaline phosphatase and adenosine deaminase in pregnant women%妊娠女性α-L-岩藻糖苷酶、碱性磷酸酶和腺苷脱氨酶活性观察

    王玮玮; 孙伟才; 周惠玉


    目的:妊娠期女性观察血清α-L-岩藻糖苷酶(AFU)、碱性磷酸酶(ALP)及腺苷脱氨酶(ADA)活性的变化及其与妊娠周期的相关性。方法分别检测352例体检正常的不同妊娠期女性(妊娠组,其中早期妊娠210例、晚期妊娠142例)及322例因不孕就诊且尚未妊娠的体检正常的女性(非妊娠组)血清 AFU、ALP、ADA 活性。对所有对象均进行动态监测(早期妊娠组于妊娠第14、22、30及38周分别检测,晚期妊娠组于妊娠第38及40周检测,非妊娠组于就诊时、就诊后第8及24周检测)并做比较。结果妊娠组血清 ALP、AFU 活性明显高于非妊娠组(P =0.000),ADA 活性则低于非妊娠组(P =0.000)。晚期妊娠组血清 ALP、AFU 活性明显高于早期妊娠组(P =0.000),但 ADA 活性两组间差异无统计学意义(P >0.05)。对早期妊娠组的动态监测显示,随着妊娠周期的增加,AFU 及 ALP 活性逐渐升高,但 ADA 基本无变化。晚期妊娠组第38周及第40周血清 AFU、ALP、ADA 活性差异均无统计学意义(P >0.05)。对非妊娠组的动态监测显示,与就诊时比较,就诊后第8周 ALP、ADA 活性差异有统计学意义(P =0.000),就诊后第24周 ADA 活性差异有统计学意义;与就诊后第8周比较,就诊后第24周 ALP、AFU、ADA 活性差异均有统计学意义(P <0.05)。结论妊娠期女性血清 AFU 及 ALP 活性明显升高,且随孕周的增加,升高趋势越明显。%Objective To observe the activity changes of alpha-L-fucosidase (AFU),alkaline phosphatase (ALP) and adenosine deaminase (ADA)in pregnant women and their correlations with pregnant period.Methods The activities of AFU,ALP and ADA were determined in 352 pregnant women (210 cases with early-stage pregnancy and 142 cases with late-stage pregnancy)and 322 unpregnant women.Dynamic monitoring was performed (early-stage pregnancy

  4. Low serum concentrations of 25-hydroxyvitamin D in children and adolescents with systemic lupus erythematosus.

    Peracchi, O A B; Terreri, M T R A; Munekata, R V; Len, C A; Sarni, R O S; Lazaretti-Castro, M; Hilário, M O E


    We evaluated the concentrations of 25-hydroxyvitamin D [25(OH)D] in children and adolescents with juvenile systemic lupus erythematosus (JSLE) and associated them with disease duration and activity, use of medication (chloroquine and glucocorticoids), vitamin D intake, calcium and alkaline phosphatase levels, and bone mineral density. Thirty patients with JSLE were evaluated and compared to 30 healthy individuals, who were age and gender matched. Assessment was performed of clinical status, disease activity, anthropometry, laboratory markers, and bone mineral density. The 30 patients included 25 (83.3%) females and 16 (53.3%) Caucasians, with a mean age of 13.7 years. The mean age at diagnosis was 10.5 years and mean disease duration was 3.4 years. Mean levels of calcium, albumin, and alkaline phosphatase were significantly lower in patients with JSLE compared with controls (Pvitamin D levels vitamin D levels between 20 and 32 ng/mL. However, these values were not associated with greater disease activity, higher levels of parathormone, medication intake, or bone mineral density. Vitamin D concentrations were similar with regard to ethnic group, body mass index, height for age, and pubertal stage. Significantly more frequently than in controls, we observed insufficient serum concentrations of 25(OH)D in patients with JSLE; however, we did not observe any association with disease activity, higher levels of parathormone, lower levels of alkaline phosphatase, use of medications, or bone mineral density alterations.

  5. Serum biochemical profile of laying hens in the region of Araçatuba, SP
    Perfil bioquímico das galinhas poedeiras na região de Araçatuba-SP.

    Paulo César Ciarlini; Elisa Helena Giglio Ponsano; Lorrayne Bernegossi Polônio; Carolina Kimie Mori; Tatiana de Sousa Barbosa


    The establishment of reference values is extremely important for successful diagnosis and treatament. Considering that in most species the serum chemistry profile is influenced by race, climate and management, we decided to determine the values of aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid, creatinine, creatine kinase (CK), phosphatase alkaline (ALP), gamma-glutamyltransferase (GGT), total protein (TP) and albumin of Dekalb hens in the region of Araçatuba - SP...

  6. 秦皇岛近海褐潮高发区浮游植物的碱性磷酸酶活性分析%Alkaline phosphatase activity of phytoplankton in Qin-huangdao coastal waters with frequent brown tide occurrences

    刘晓红; 覃仙玲; 蔡阳扬; 欧林坚; 吕颂辉


    Alkaline phosphatase (AP) is a type of ectoenzyme that is expressed by phytoplankton under phosphorus (P) stress states and can hydrolyze dissolved organic P (DOP). AP has been widely used to indicate the P stress status of phytoplankton in the sea. In this study, the AP activity (APA) of phytoplankton during the brown tide of Aureococcus anophagefferens that occurred near the coastal waters of Qinghuangdao was investigated in July 2013. The P status of phytoplankton and the capability of phytoplankton in using DOP during the bloom were analyzed. The results show that the density of A. anophagefferens was as high as 108 cells/L when the bloom occurred in the coastal area of Qinhuangdao. DOP was the main P source for phytoplankton growth. The densities of A. ano-phagefferens showed positive correlations with NO3−, DOP, dissolved inorganic P, and so forth. Phytoplankton ex-pressed abundant AP, and the average APA in water reached as high as 217.72 nmol/(µg·h)chla ± 90.86 nmol/(µg·h)chla (350.44 nmol/(L·h) ± 130.57 nmol/(L·h)). APA increased significantly with phytoplankton biomass. Our results suggest that A. anophagefferens experienced severe P stress or even P limitation when the brown tide occurred. The bioavailability of P, particularly DOP, may play a key role in the occurrence and persistence of the brown tide of A. anophagefferens in the coastal waters of Qinhuangdao.%碱性磷酸酶(Alkaline phosphatase, AP)是浮游植物在磷胁迫状态下表达的一种水解有机磷源的胞外酶,可用于指示海区浮游植物的磷胁迫状态。本研究于2013年7月,对秦皇岛近海抑食金球藻(Aureococcus anophagefferens)褐潮发生期间浮游植物的碱性磷酸酶活性(AP activity, APA)进行研究,结合其他理化参数,分析藻华发生时浮游植物的磷营养状态及其对海水中磷源的水解与利用情况。结果表明,褐潮发生时,抑食金球藻细胞密度高达108个/L,溶解有机磷(Dissolved organic phosphorus

  7. 泥鳅仔稚鱼发育期间消化酶及碱性磷酸酶比活力的变化%Changes of digestive enzymes and alkaline phosphatase specific activities in Misgurnus anguillicaudatus during the development of early larval ontogeny

    张云龙; 樊启学; 彭聪; 胡培培; 宗克金; 宋林; 沈凡


    The specific activities of several digestive enzymes (pensin, trypsin, lipase, amylase) and alkaline phosphatase of Misgurnus anguillicaudatus were determined from hatching to 30 DAH ( days after hatching). Pepsin activity could not be detected during the entire period of the experiment. In contrast, trypsin performed a high level activity after hatching , and its specific activity increased significantly after first feeding, peaking at 6 DAH, and decreased abruptly after that. The variation of lipase and amylase shared a similar pattern, their specific activities had two obvious peaks around the time of the transition of endogenous to exdogenous feeding and the beginning of the transportation of larval stage to juvenile stage. From 2 to 6 DAH, the specific activity of alkaline phosphatase significantly increased and then declined to a roughly stable level. This study demonstrated that M. anguillicaudatus has only a structural not functional stomach during the larval stage. The period of 2 to 6 DAH was the stage of the maturation of digestive gut in M. anguillicaudatus larvae and the crucial step forward the digestion mode of adult M. anguillicaudatus. The steadiness of these specific activities suggested that M. anguillicaudatus was efficient to utilize carbohydrate and lipid in food.%研究了泥鳅(Misgurnus anguillicaudatus)从孵化至30 DAH(日龄,Days after hatching)几种消化酶及碱性磷酸酶比活力的变化情况.胃蛋白酶直至30 DAH仍未检出活性.而胰蛋白酶表现出较高的比活力,其比活力在初次摄食之后显著上升,6 DAH达到最大值之后开始显著降低(P<0.05).脂肪酶与淀粉酶的变化模式相似,在内源性营养向外源性营养转变及仔鱼向稚鱼转变这两个时间段出现两个高峰值.碱性磷酸酶比活力在2-6 DAH显著上升(P<0.05),之后开始下降并趋于平稳.研究表明,泥鳅在仔稚鱼阶段只具有结构性的胃而缺乏分泌细胞的分化.2-6 DAH是泥鳅仔鱼肠

  8. Alkaline Phosphatase Activity and Ⅰ Collagen Expression of Rat Dental Papilla Cells in Virto%体外诱导培养大鼠牙乳头细胞碱性磷酸酶和Ⅰ型胶原的表达

    曹莹; 方厂云; 吕亚林; 姚志刚


    目的:观察体外诱导培养的大鼠牙乳头细胞碱性磷酸酶和Ⅰ型胶原蛋白合成的变化.方法:原代培养法获得大鼠牙乳头细胞,在条件培养基中进行诱导培养,在不同时间进行钙结节染色,ALP染色和定量测定,ELISA法和RT-PCR法测定细胞Ⅰ型胶原蛋白的量和mRNA表达水平.结果:体外诱导培养的大鼠牙乳头细胞形成含钙化物的细胞结节,体外培养至12 d的诱导组细胞ALP高于对照组(P< 0.05),细胞内1型胶原表达量高于对照组(P<0.05),且Ⅰ型胶原mRNA表达水平上调(P<0.05);培养15d后诱导组细胞分泌的Ⅰ型胶原高于对照组(P<0.05).结论:大鼠牙乳头细胞在体外分化的过程中碱性磷酸酶活性明显增高,矿化能力增强,并不断合成和分泌Ⅰ型胶原形成胞外基质.%Objective: To investigate the alkaline phosphatase (ALP ) activity and type I collagen content of rat dental papillae cells (rDPC) cultured in odontoblast differentiation-inducing medium in vitro. Methods: Neonatal rat dental papillae cells were obtained from molar tooth germs and expanded in vitro. rDPC were cultured in the medium containing dexamethasone, ascorbic acid and β-glycerophosphate. Cell cultures were analysed for morphology, mineralization potential, alkaline phosphatase and Type I collagen using immunocytochemistry, ELISA and reverse transcriptase-polymerase chain reaction. Results: Cell mineral nodules of rDPC cultured in odontoblast differentiation- inducing medium were detect by Alizarin red S staining after day 7. ALP activity, type I collagen in cell culture supernatants and type I collagen mRNA expression of rDPC were all increased significantly compared with control group after day 12. Type I collagen concentrations in intracellular protein of differentiation-inducing cultured rDPC were significantly enhanced compared to rDPC in control group after day 15. Conclusion: Odontoblast differentiation-inducing treatment promoted the

  9. Influence of triethyl phosphate on phosphatase activity in shooting range soil: Isolation of a zinc-resistant bacterium with an acid phosphatase.

    Story, Sandra; Brigmon, Robin L


    Phosphatase-mediated hydrolysis of organic phosphate may be a viable means of stabilizing heavy metals via precipitation as a metal phosphate in bioremediation applications. We investigated the effect of triethyl phosphate (TEP) on soil microbial-phosphatase activity in a heavy-metal contaminated soil. Gaseous TEP has been used at subsurface sites for bioremediation of organic contaminants but not applied in heavy-metal contaminated areas. Little is known about how TEP affects microbial activity in soils and it is postulated that TEP can serve as a phosphate source in nutrient-poor groundwater and soil/sediments. Over a 3-week period, TEP amendment to microcosms containing heavy-metal contaminated soil resulted in increased activity of soil acid-phosphatase and repression of alkaline phosphatase, indicating a stimulatory effect on the microbial population. A soil-free enrichment of microorganisms adapted to heavy-metal and acidic conditions was derived from the TEP-amended soil microcosms using TEP as the sole phosphate source and the selected microbial consortium maintained a high acid-phosphatase activity with repression of alkaline phosphatase. Addition of 5mM zinc to soil-free microcosms had little effect on acid phosphatase but inhibited alkaline phosphatase. One bacterial member from the consortium, identified as Burkholderia cepacia sp., expressed an acid-phosphatase activity uninhibited by high concentrations of zinc and produced a soluble, indigo pigment under phosphate limitation. The pigment was produced in a phosphate-free medium and was not produced in the presence of TEP or phosphate ion, indicative of purple acid-phosphatase types that are pressed by bioavailable phosphate. These results demonstrate that TEP amendment was bioavailable and increased overall phosphatase activity in both soil and soil-free microcosms supporting the possibility of positive outcomes in bioremediation applications.

  10. Measurement and analysis of alkaline phosphatase and aspartate aminotransferase activity levels in gingival cervical fluid after treatment of telescope retained fixed bridge%套筒冠式固定桥基牙龈沟液中碱性磷酸酶和天冬氨酸转氨酶的检测分析

    武峰; 赵彬; 姚蔚


    Objective To explore the role of telescopic crown retainers fixed bridge in periodontal maintenance. Methods A total of 64 abutment teeth were selected from 16 patients and divided into experimental and control groups (n=32 each). The gingival index and sulcus bleeding index of abutments in the both two groups were detected at 6 and 12 months before and after repair. The alkaline phosphatase (ALP) and aspartate aminotransferase (AST) activity levels in gingival cervical fluid were measured using automatic biochemistry analyzer. Results There were significant differences in Gl and SBI between the experimental and control groups, as well as in ALP and AST activity levels (P<0.05). Conclusion Telescope retained fixed bridge might help protect the periodontal tissue of abutment teeth.%目的 探索套筒冠固位式固定桥对于牙周维护的作用.方法 选择16例患者共64颗基牙,分为实验组和对照组,每组32颗基牙.在修复前以及修复后6 、12个月,对基牙进行牙龈指数(GI)和龈沟出血指数(SBI)的检测,利用全自动生化分析仪检测基牙龈沟液中碱性磷酸酶(ALP)和天冬氨酸转氨酶(AST)活性水平.结果 在修复前与修复后12 个月实验组与对照组之间GI、SBI比较差异有统计学意义(P<0.05),ALP、AST活性水平差异也有统计学意义(P<0.05).结论 套筒冠式固定桥有利于基牙牙周组织的保护.

  11. Serum fibroblast growth factor 23, serum iron and bone mineral density in premenopausal women.

    Imel, Erik A; Liu, Ziyue; McQueen, Amie K; Acton, Dena; Acton, Anthony; Padgett, Leah R; Peacock, Munro; Econs, Michael J


    Fibroblast growth factor 23 (FGF23) circulates as active protein and inactive fragments. Low iron status increases FGF23 gene expression, and iron deficiency is common. We hypothesized that in healthy premenopausal women, serum iron influences C-terminal and intact FGF23 concentrations, and that iron and FGF23 associate with bone mineral density (BMD). Serum iron, iron binding capacity, percent iron saturation, phosphorus, and other biochemistries were measured in stored fasting samples from healthy premenopausal white (n=1898) and black women (n=994), age 20-55years. Serum C-terminal and intact FGF23 were measured in a subset (1631 white and 296 black women). BMD was measured at the lumbar spine and femur neck. Serum phosphorus, calcium, alkaline phosphatase and creatinine were lower in white women than black women (piron (piron. C-terminal FGF23 correlated inversely with iron (white women r=-0.134, piron iron predicted changes in C-terminal FGF23. Spine BMD correlated with iron negatively (r=-0.076, piron negatively (r=-0.119, piron did not relate to intact FGF23, but was inversely related to C-terminal FGF23. Intact FGF23 correlated with serum phosphorus. In weight-adjusted models, BMD was not related to intact FGF23, C-terminal FGF23 or iron. The influence of iron on FGF23 gene expression is not important in determining bone density in healthy premenopausal women.

  12. Effect of Heat Stress on the Intestinal Flora Structure and Alkaline Phosphatase Activities and mRNA Expression of Amino Acid Transporters of Layer%热应激对蛋鸡肠道菌群结构、碱性磷酸酶活性及氨基酸转运载体mRNA表达丰度的影响

    李永洙; 陈常秀; Yongquan Cui


    [目的]揭示热应激环境下蛋鸡肠道菌群结构、肠黏膜碱性磷酸酶活性和氨基酸转运载体mRNA表达量的变化机理。[方法]试验选择16周龄济宁百日鸡96只,随机分成对照组((24±1)℃;Ⅰ)和热应激((38±1)℃)组,各组设6个重复,每个重复8只,试验持续14 d。采用16S rDNA的PCR-DGGE技术和实时荧光定量PCR等手段,分析热应激2(Ⅱ)、7(Ⅲ)、14 d(Ⅳ)时,对十二指肠、空肠及回肠内容物菌群多样性和肠黏膜碱性磷酸酶活性以及氨基酸转运载体rBAT、y+LAT 1、CAT l mRNA基因表达的相对丰度变化规律。[结果]热应激7 d开始各肠段菌群多样性较为丰富,热应激7、14 d时空肠和回肠部位敏感乳杆菌(Lactobacillus agilis),回肠部位约氏乳杆菌(Lactobacillus johnsonii)、不可培养细菌(uncultured bacterium)和不可培养的拟杆菌属(uncultured Bacteroidalesbacterium)均末检测到;而热应激不同时间段空肠和回肠部位可检测到不可培养细菌、溃疡拟杆菌(Bacteroides helcogenes)、卵形拟杆菌(Bacteroides ovatus)、索氏志贺氏菌(Shigella sonnei);空肠和回肠部位黏膜上皮细胞表面的碱性磷酸酶活性与Ⅰ组比较显著下降(P<0.05);而空肠和回肠Ⅲ组的rBAT、y+LAT 1 mRNA表达丰度均最低,空肠在各热应激时段表达丰度变化幅度最大(P<0.05),回肠的CAT 1 mRNA表达丰度在Ⅲ、Ⅳ组与Ⅰ组比较影响更明显(P<0.01)。[结论]热应激对空肠和回肠部位微生物菌群影响较为明显,肠道微生物群落改变可导致肠道的消化吸收功能发生改变。%[Objective] The objective of this study is to reveal the influence mechanisms of heat stress affecting the intestinal flora structure of layer, the alkaline phosphatase activities of intestinal mucosa and the mRNA expression of amino acid transporters.[Method]A total of 96 Jining Bairi

  13. Dephosphorylation of Centrins by Protein Phosphatase 2C and

    Marie-Christin Thissen


    Full Text Available In the present study, we identified protein phosphatases dephosphorylating centrins previously phosphorylated by protein kinase CK2. The following phosphatases known to be present in the retina were tested: PP1, PP2A, PP2B, PP2C, PP5, and alkaline phosphatase. PP2C and were capable of dephosphorylating P-Thr138-centrin1 most efficiently. PP2C was inactive and the other retinal phosphatases also had much less or no effect. Similar results were observed for centrins 2 and 4. Centrin3 was not a substrate for CK2. The results suggest PP2C and to play a significant role in regulating the phosphorylation status of centrins in vivo.




    Full Text Available The purpose of this study was to determine a simple and sensitive test for clinical diagnosis of various hepatic diseases. Therefore y-glutamyltranspeptidase (Y** GT, aspartate aminotransferase (ASAT, alanine aminotr¬ansferase (ALAT and alkaline phosphatase (ALP levels were measured in 29 healthy adults and 88 sera with various liver diseases. Table I represents the results, according to which y-GT activity increases in all of studied patients, especially in alcoholic liver disease and hepatobiliary dysfunction (13, 5, 3,10, 4."nThe data suggest that in liver disease it is better to estimate y-GT level in serum prior to other related enzymes.

  15. Direct determination of phosphatase activity from physiological substrates in cells.

    Zhongyuan Ren

    Full Text Available A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min(-1 mg(-1 for PPi, to 56 ± 11 nmol min(-1 mg(-1 for AMP, to 79 ± 23 nmol min(-1 mg(-1 for beta-glycerophosphate and to 73 ± 15 nmol min(-1 mg(-1 for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole--a TNAP inhibitor--served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes.

  16. Multiple myeloma: Changes in serum C-terminal telopeptide of collagen type I and bone-specific alkaline phosphatase can be used in daily practice to detect imminent osteolysis

    Lund, Thomas; Abildgaard, Niels; Andersen, Thomas L;


    Abstract Objective: Monitoring of bone disease in multiple myeloma is becoming increasingly important since bone protecting treatment with bisphosphonate is becoming restricted after the awareness of osteonecrosis of the jaw. Despite the potential of biochemical markers of bone remodeling...

  17. Serum Lp(a)levels in patients with liver disease

    Sharanabasappa M; Sudhakar Nayak


    Objective:The study was conducted to evaluate the effect of liver diseases on serum Lp(a)levels and also to study the relationship between Lp(a)levels with other lipid parameters and liver function tests in 32 hyperbil-irubinemia patients with total bilirubin >3mg/dL.The results were compared with 20 healthy age matched in-dividuals.Methods:Serum obtained from venous blood sample are used for estimating total cholesterol (TC), triglyceride(TG),high density lipoprotein cholesterol(HDL-c),low density lipoprotein cholesterol(LDL-c), very low density lipoprotein cholesterol(VLDL-c),total protein(TP),albumin(ALB),total bilirubin,direct bilirubin,aspartate amino tranferase(AST),alanine amino transferase(ALT),alkaline phosphatase(ALP), gamma glutamyl transferase (GGT),lipoprotein (a)[Lp(a)],serum phospholipids.Results:There was significant decrease in serum Lp(a)levels in liver disease patients and the decrease was directly correlated with reduced serum albumin levels and inversely with liver function parameters.Conclusion:Thus the present study indicates hepatic synthetic damage has possible biochemical basis for the reduction of serum Lp(a)lev-els.

  18. Alkaline phosphatases. Newer concepts in isoenzymes and clinical applications.

    Griffiths, J


    ALP is a profoundly ubiquitous enzyme that originates in many, if not all, mammalian tissues. The development of a simple ISEF technique has allowed reflection of at least 12 cellular components. Improvements in technique may add further zones of information. The aura of darkness surrounding ALP is being slowly dissipated.

  19. Serum enzymes levels and influencing factors in three indigenous Ethiopian goat breeds.

    Tibbo, M; Jibril, Y; Woldemeskel, M; Dawo, F; Aragaw, K; Rege, J E O


    Serum enzymes were studied in 163 apparently healthy goats from three indigenous goat breeds of Ethiopia. The effect of breed, age, sex and season on alanine aminotransferase (ALT) / glutamic pyruvic transaminase (GPT), aspartate aminotransferase (AST) / glutamic oxalacetic transaminases (GOT), alkaline phosphatase (ALP) and acid phosphatase (AcP) levels was assessed. The mean serum enzymes levels of the indigenous Arsi-Bale, Central Highland and Long-eared Somali goat breeds ranged from 14.0-20.2 iu L(-1) for ALT/GPT, from 43.2-49.3 iu L(-1) for AST/GOT, from 83.7-98.8 iu L(-1) for ALP, and from 2.99-4.23 iu L(-1) for AcP, were within the normal range for goats elsewhere. Breed had significant influence on AST/GOT values. Sex had significant effect on ALT/GPT for Arsi-Bale goats with higher values in males than females. Age was significant on all serum enzymes studied in the Arsi-Bale goats and on ALP in the Central Highland goats. Season had significant influence on all serum enzymes except for ALT/GPT in the Arsi-Bale goats. The serum enzyme levels of these indigenous goat breeds can be used as normal reference values for Ethiopian goat breeds adapted to similar agro-ecology and production system.

  20. Blood serum components and serum protein test of Hybro-PG broilers of different ages

    PRL Silva


    Full Text Available Blood serum samples of HYBRO PG broilers were analyzed, with 30 samples collected from 21-day-old broilers (G1, 30 from 35-day-old birds (G2, and 30 from 42-day-old birds (G3, with the aim of establishing normal values of some blood serum parameters. The activities of the enzymes gamma-glutamyl-transferase (GGT, aspartate aminotransferase (AST, creatine kinase (CK, alkaline phosphatase (ALP, and lactate dehydrogenase (LDH, serum levels of total calcium, calcium ion, phosphorus, sodium, potassium, magnesium, chlorides, creatinine, uric acid, triglycerides, cholesterol, total protein, albumin, total and indirect and direct bilirubin, and electrophoretic profile of serum proteins in acrylamide (SDS-PAGE and agarose gel were determined. There was no influence of age on total bilirubin and albumin levels. All the other evaluated parameters presented differences in at least one age group. Protein electrophoretic profile also changed as a function of age. The obtained results can be considered as normal for the studied ages, and therefore be used as references for the interpretation of laboratory exams of broilers of this genetic line in the evaluated ages.

  1. Serum osteoprotegerin (OPG in children with primary nephrotic syndrome

    Gamal B Mohamed


    Full Text Available A novel cytokine system secreted by osteoblast, osteoprotegerin (OPG and its ligand (OPGL regulates osteoclastogenesis. To determine the relation of the serum OPG levels in children with nephrotic syndrome (NS to the renal disease, we studied 30 patients with NS in comparison with 30 healthy children serving as controls. The study patients were divided into three equal groups: group 1 included newly diagnosed patients who were studied before and after a short course (one month of steroid therapy for the first time, group 2 included frequent relapsers (FR, and group 3 included infrequent relapsers (IFR. In addition to serum OPG (ELISA, osteocalcin (OC, parathormone (PTH, alkaline phosphatase (ALP, and 24- hour urinary Ca and proteins were measured. The NS patients revealed a significantly lower serum OPG and parameters of bone formation (ALP and OC and a significantly higher 24- hour urinary Ca than controls. A short course of glucocorticoids therapy for one month resulted in a significant decrease of serum OPG, ALP and OC levels and a significant increase of 24- hour urinary Ca, while serum PTH levels were not significantly affected by this the- rapy; the FR revealed a significantly lower serum level and a significantly higher 24- hour urinary Ca and serum PTH than the IFR. OPG had significant negative correlations with markers of disease activity and severity (ESR, serum cholesterol, 24- hour urinary protein and cumulative steroid dose, PTH and 24- hour urinary Ca. On the other hand, OPG had significant positive correlations with ALP, OC, and serum albumin. Low serum OPG, which is attributed to the renal disease and/or steroid therapy, may be an important factor contributing to bone resorption in NS. Studies of the protective effect of OPG administration against bone loss in NS are warranted.

  2. Expression of bacterial alkaline phosphatase-steroid receptor coactivator-1 fusion protein and its application in regulation of drug metabolism enzymes%BAP-SRC-1融合蛋白的表达及其在药物代谢酶调控研究中的应用

    陈亚坤; 陈枢青; 曾苏


    目的获得活性表达的细菌碱性磷酸酶(bacterial alkaline phosphatase,BAP)基因和人甾体受体辅活化因子-1(stero1d receptor coactivator-1,SRC-1)的融合蛋白,应用于辅活化因子与核受体的结合研究.方法从Escherichia coli JM83基因组和人肝总RNA中分别扩增获得BAP基因和SRC-1的186个氨基酸对应的基因序列(简称SRC186).用重组技术,构建BAP-SRC186-pET28a融合基因表达载体,在Escherichia coli Rosetta(DE3)中,以IPTG低温诱导表达.以对硝基苯磷酸盐(p-nitrophenyl-phos-phate,PNPP)为底物进行活性测定.活性表达的BAP-SRC186融合蛋白被应用于辅活化因子与核受体的结合研究.结果获得可溶性融合蛋白BAP-SRC186.该融合蛋白的BAP比活为(0.176±0.013 4)μmol·min-1·mg(pro).在利福平存在的情况下,BAP-SRC186能与孕烷X受体配体结合域(pregnane X receptor ligand binding domain,PXRLBD)发生利福平剂量依赖性的相互作用,作用强度通过BAP的显色反应可方便地检测.结论 BAP融合蛋白与核受体的结合研究为药物代谢酶调控的体外研究开辟了新的思路.

  3. Clinical significance of γ-glutamyl transferase,alkaline phosphatase,high-sensitivity C-reactive protein in elderly patients with subtrochanteric fractures%老年股骨粗隆骨折患者γ-谷氨酰转肽酶、碱性磷酸酶、超敏C反应蛋白联合检测的临床意义

    王涵; 叶玉芬; 周建球; 姚荣忠; 谭放花


    目的 探讨老年股骨粗隆骨折患者γ-谷氨酰转移酶(GGT)、碱性磷酸酶(ALP)、超敏C反应蛋白(hs-CRP)联合检测的临床意义.方法 回顾性分析67例老年股骨粗隆骨折患者的GGT、ALP、hs-CRP水平,并研究术后不同时间点各指标相对于术前的变化情况及不同降低水平的预后情况.结果 老年股骨粗隆骨折患者的GGT、ALP及hs-CRP水平高于健康对照者和其他部位骨折患者,且与病理参数有关;术后呈降低的趋势;老年股骨粗隆骨折患者中三项指标术后水平降幅较大者的Harris髋关节功能评分累积优良率高于降幅者低者,差异有统计学意义(P<0.01).结论 监测老年股骨粗隆骨折患者的血清GGT、ALP及hs-CRP水平及其术后的变化情况具有重要的临床意义.%Objective To explore the clinical significance of y glutamyl transferase(GGT) ,alkaline phosphatase(ALP) and high sensitivity C reactive protein(hs CRP) detection in elderly patients with subtrochanteric fractures. Methods GGT, ALP and hs CRP levels in 67 elderly patients with subtrochanteric fractures were retrospectively analyzed,and the changes of them before oper ation and at different postoperative time points as well as the prognosis of reducing level were also analyzed. Results GGT,ALP and hs CRP levels in patients with subtrochanteric fractures were higher than in health people and patients with other fractures,and were related to the pathological parameters. The three indicators showed a decreasing trend after operation. The Harris hip score ac cumulated good rates in high level decreasing groups were higher than those in low level decreasing groups(P<0. 01). Conclusion GGT,ALP and hs CRP levels and changes before and after surgery in elderly patients with subtrochanteric fractures might be with important clinical significance.

  4. Expression cloning of different bacterial phosphatase-encoding genes by histochemical screening of genomic libraries onto an indicator medium containing phenolphthalein diphosphate and methyl green.

    Riccio, M L; Rossolini, G M; Lombardi, G; Chiesurin, A; Satta, G


    A system for expression cloning of bacterial phosphatase-encoding genes has been developed, and its potential has been investigated. The system is based on histochemical screening of bacterial genomic libraries, constructed in an Escherichia coli multicopy plasmid vector, for phosphatase-producing clones using an indicator medium (named TPMG) made of Tryptose-Phosphate agar supplemented with the phosphatase substrate phenolphthalein diphosphate and the stain methyl green. To test the performance of this system, three genomic libraries were constructed from bacterial strains of different species which showed different patterns of phosphatase activity, and were screened using the TPMG medium. Following a partial screening, three different phosphatase-encoding genes (respectively encoding a class A non-specific acid phosphatase, an acid-hexose phosphatase and a non-specific alkaline phosphatase) were shotgun-cloned from the above libraries, indicating that the TPMG-based expression cloning system can be useful for rapid isolation of different bacterial phosphatase-encoding genes.

  5. Serum chemical values in hypothermic and rewarmed young calves.

    Olson, D P; South, P J; Hendrix, K


    Serum chemical values were determined in cold-stressed Holstein bull calves ranging from 1 to 7 days of age. The animals were anesthetized and cold-stressed until their core body temperature (colonic) was lowered 10 C. Animals were then rewarmed in warm water, with heat pads or heat lamps, or were allowed to recover naturally (unassisted) at room temperature. Blood samples were collected at selected intervals during cooling and recovery. Increases (P less than 0.05) were observed in the concentrations of glucose, calcium, phosphorus, iron, alkaline phosphatase, aspartate aminotransferase, lactate dehydrogenase, total protein, albumin, total globulin, serum urea nitrogen, uric acid, total bilirubin, indirect bilirubin, and cholesterol in the cold-stressed calves during cooling. Concentrations of chloride and insulin decreased (P less than 0.05) during the same period. Changes observed in many of the serum chemical values during rewarming were generally the reverse of the respective changes that occurred during cooling, although insulin values became exceedingly high in some cases midway or near the end of recovery. Serum enzyme values also remained high during most of recovery. Data did not indicate a clear advantage of one method of rewarming over the other methods used in terms of return of the serum chemical values to normal.

  6. [Effect of low-intensity 900 MHz frequency electromagnetic radiation on rat liver and blood serum enzyme activities].

    Nersesova, L S; Petrosian, M S; Gazariants, M G; Mkrtchian, Z S; Meliksetian, G O; Pogosian, L G; Akopian, Zh I


    The comparative analysis of the rat liver and blood serum creatine kinase, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and purine nucleoside phosphorylase post-radiation activity levels after a total two-hour long single and fractional exposure of the animals to low-intensity 900 MHz frequency electromagnetic field showed that the most sensitive enzymes to the both schedules of radiation are the liver creatine kinase, as well as the blood serum creatine kinase and alkaline phosphatase. According to the comparative analysis of the dynamics of changes in the activity level of the liver and blood serum creatine kinase, alanine aminotransferase, aspartate aminotransferase and purine nucleoside phosphorylase, both single and fractional radiation schedules do not affect the permeability of a hepatocyte cell membrane, but rather cause changes in their energetic metabolism. The correlation analysis of the post-radiation activity level changes of the investigated enzymes did not reveal a clear relationship between them. The dynamics of post-radiation changes in the activity of investigated enzyme levels following a single and short-term fractional schedules of radiation did not differ essentially.

  7. Serum osteocalcin is associated with dietary vitamin D, body weight and serum magnesium in postmenopausal women with and without significant coronary artery disease.

    Alissa, Eman M; Alnahdi, Wafa A; Alama, Nabeel; Ferns, Gordon A


    Osteoporosis and atherosclerosis often present atypically in postmenopausal women, making clinical recognition difficult. Prospective studies suggest independent associations between bone mass and vascular calcification through vitamin D deficiency as an established predictor of both conditions. We aimed to examine the relationship between serum osteocalcin and vitamin D status in postmenopausal women with and without angiographic evidence of coronary artery disease (CAD). One hundred and eighty postmenopausal women undergoing coronary angiography were selected sequentially from the Catheterization unit of King Abdulaziz University Hospital. Socio-demographic, anthropometric parameters and dietary habits were measured. Biochemical variables were estimated in blood samples. Half of the postmenopausal women did not have significant CAD, 24% had significant CAD in a single and/or double coronary vessels, 26% had significant CAD in three coronary vessels. Mean serum vitamin D concentrations showed that vitamin D deficiency was a common finding in the whole population. Vitamin D and calcium intakes were uniformly low in the study cohort. Serum osteocalcin was significantly correlated with dietary vitamin D in all subgroups (r=-0.172, pmagnesium, alkaline phosphatase, dietary vitamin D, and body weight were independent variables of serum osteocalcin level. In conclusion, elevated levels of serum C reactive protein and vitamin D were associated with low serum osteocalcin levels. Therefore, osteocalcin may be a potential cardiovascular risk marker. However, further studies are needed to clarify the pathophysiological processes underlying the relationship between serum osteocalcin level and atherosclerosis parameters.

  8. Glucose-6-phosphatase deficiency.

    Labrune Philippe; Gajdos Vincent; Eberschweiler Pascale; Hubert-Buron Aurélie; Petit François; Vianey-Saban Christine; Boudjemline Alix; Piraud Monique; Froissart Roseline


    Abstract Glucose-6-phosphatase deficiency (G6P deficiency), or glycogen storage disease type I (GSDI), is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, betw...

  9. Comparison of alkaline phosphatase in Bemisia tabaci B-biotype(Homoptera: Aleyrodidae) and Trialeurodes vaporariorum (Homoptera:Aleyrodidae) at different developmental stages%B型烟粉虱与温室白粉虱不同虫态的碱性磷酸酶性质比较

    严盈; 刘万学; 万方浩


    为了探明B型烟粉虱,Bemisia tobaci B-biotype和温室白粉虱Trialeurodes vaporariorum体内的碱性磷酸酶(alkaline phosphatase,ALP)在两者竞争替代中所起的作用,以对硝基苯磷酸二钠(pNPP)为底物,采用个体测定和群体测定的方法,研究比较了2种粉虱不同虫态中该酶的性质.结果表明:2种粉虱的碱性磷酸酶比活力在整个发育历期均逐渐增加,成虫期达到最大.温室白粉虱2至4龄若虫(伪蛹)期的碱性磷酸酶比活力分别是B型烟粉虱对应龄期酶比活力的2.58、2.68和3.14倍;B型烟粉虱雌雄成虫的碱性磷酸酶比活力分别是温室白粉虱雌雄成虫酶比活力的1.24和1.26倍,且2种粉虱雌虫的酶比活力显著大于其雄虫.2种粉虱2龄若虫到成虫的碱性磷酸酶最适pH均为7.8,最适温度均为47 ℃;在1龄若虫中均未能检测到该酶活性.测定并比较2种粉虱不同虫态碱性磷酸酶动力学特征参数的结果显示,温室白粉虱碱性磷酸酶在3、4龄若虫的亲和力以及在2,3,4龄若虫的酶蛋白浓度均显著大于B型烟粉虱的对应值,而在成虫期2种粉虱的亲和力、酶蛋白浓度无差异,B型烟粉虱的活化能显著小于温室白粉虱.据此推测,B型烟粉虱利用碱性磷酸酶在若虫期进行组织骨化和生长发育不如温室白粉虱,但羽化为成虫后利用其进行解毒代谢则可能强于温室白粉虱.

  10. Research on Phosphatases of Belladona Leaves and Their Purification (Part 1

    M. Khorsand


    Full Text Available Belladona leaves as well as all other studied leaves contains two distinct phosphatase fractions belonging respectively to types II and IIIi the major parts of these enzymes is extraetible by water. It was not possible to extract the non soluble fraction which is solidly retained by the cellular constituents. Phosphatase II does not differ from other phosphatnses of the same type. Whereas phosphatase III is distinetely different from enzymes of the same type of vegetal or animal origins. It is activated by bivalent metallic ions which are specific activators of the alkaline phcspbatnses: Mg-Zn-Ni and Co.

  11. Function-Based Metagenomic Library Screening and Heterologous Expression Strategy for Genes Encoding Phosphatase Activity.

    Villamizar, Genis A Castillo; Nacke, Heiko; Daniel, Rolf


    The release of phosphate from inorganic and organic phosphorus compounds can be mediated enzymatically. Phosphate-releasing enzymes, comprising acid and alkaline phosphatases, are recognized as useful biocatalysts in applications such as plant and animal nutrition, bioremediation and diagnostic analysis. Metagenomic approaches provide access to novel phosphatase-encoding genes. Here, we describe a function-based screening approach for rapid identification of genes conferring phosphatase activity from small-insert and large-insert metagenomic libraries derived from various environments. This approach bears the potential for discovery of entirely novel phosphatase families or subfamilies and members of known enzyme classes hydrolyzing phosphomonoester bonds such as phytases. In addition, we provide a strategy for efficient heterologous phosphatase gene expression.

  12. [Phosphatase activity in Amoeba proteus at low pH].

    Sopina, V A


    In free-living Amoeba proteus (strain B), three forms of tartrate-sensitive phosphatase were revealed using PAGE of the supernatant of ameba homogenates obtained with 1% Triton X-100 or distilled water and subsequent staining of gels with 2-naphthyl phosphate as substrate (pH 4.0). The form with the highest mobility in the ameba supernatant was sensitive to all tested phosphatase activity modulators. Two other forms with the lower mobilities were completely or significantly inactivated not only by sodium L-(+)-tartrate, but also by L-(+)-tartaric acid, sodium orthovanadate, ammonium molybdate, EDTA, EGTA, o-phospho-L-tyrosine, DL-dithiotreitol, H2O2, 2-mercaptoethanol, and ions of heavy metals - Fe2+, Fe3+, and Cu2+. Based on results of inhibitory analysis, lysosome location in the ameba cell, and wide substrate specificity of these two forms, it has been concluded that they belong to nonspecific acid phosphomonoesterases (AcP, EC This AcP is suggested to have both phosphomonoesterase and phosphotyrosyl-protein phosphatase activitis. Two ecto-phosphatases were revealed in the culture medium, in which amebas were cultivated. One of them was inhibited by the same reagents as the ameba tartrate-sensitive AcP and seems to be the AcP released into the culture medium in the process of exocytosis of the content of food vacuoles. In the culture medium, apart from this AcP, another phosphatase was revealed, which was not inhibited by any tested inhibitors of AcP and alkaline phosphatase. It cannot be ruled out that this phosphatase belong to the ecto-ATPases found in many protists; however, its ability to hydrolyze ATP has not yet been proven.

  13. Low serum concentrations of 25-hydroxyvitamin D in children and adolescents with systemic lupus erythematosus

    O.A.B. Peracchi


    Full Text Available We evaluated the concentrations of 25-hydroxyvitamin D [25(OHD] in children and adolescents with juvenile systemic lupus erythematosus (JSLE and associated them with disease duration and activity, use of medication (chloroquine and glucocorticoids, vitamin D intake, calcium and alkaline phosphatase levels, and bone mineral density. Thirty patients with JSLE were evaluated and compared to 30 healthy individuals, who were age and gender matched. Assessment was performed of clinical status, disease activity, anthropometry, laboratory markers, and bone mineral density. The 30 patients included 25 (83.3% females and 16 (53.3% Caucasians, with a mean age of 13.7 years. The mean age at diagnosis was 10.5 years and mean disease duration was 3.4 years. Mean levels of calcium, albumin, and alkaline phosphatase were significantly lower in patients with JSLE compared with controls (P<0.001, P=0.006, and P<0.001, respectively. Twenty-nine patients (97% and 23 controls (77% had 25(OHD concentrations lower than 32 ng/mL, with significant differences between them (P<0.001. Fifteen patients (50% had vitamin D levels <20 ng/mL and 14 had vitamin D levels between 20 and 32 ng/mL. However, these values were not associated with greater disease activity, higher levels of parathormone, medication intake, or bone mineral density. Vitamin D concentrations were similar with regard to ethnic group, body mass index, height for age, and pubertal stage. Significantly more frequently than in controls, we observed insufficient serum concentrations of 25(OHD in patients with JSLE; however, we did not observe any association with disease activity, higher levels of parathormone, lower levels of alkaline phosphatase, use of medications, or bone mineral density alterations.

  14. Biomarkers of Prostatic Cancer: An Attempt to Categorize Patients into Prostatic Carcinoma, Benign Prostatic Hyperplasia, or Prostatitis Based on Serum Prostate Specific Antigen, Prostatic Acid Phosphatase, Calcium, and Phosphorus.

    Sarwar, Shahana; Adil, Mohammed Abdul Majid; Nyamath, Parveen; Ishaq, Mohammed


    Prostatitis, BPH, and P.Ca are the most frequent pathologies of the prostate gland that are responsible for morbidity in men. Raised levels of PSA are seen in different pathological conditions involving the prostate. PAP levels are altered in inflammatory or infectious or abnormal growth of the prostate tissue. Serum calcium and phosphorus levels were also found to be altered in prostate cancer and BPH. The present study was carried out to study the levels of PSA, PAP, calcium, and phosphorus in serum of patients with Prostatitis, BPH, or P.Ca and also to evaluate the relationship between them. Males in the age group of 50-85 years with LUTS disease symptoms and with PSA levels more than 4 ng/mL were included. A total of 114 patients were analyzed including 30 controls. Prostatitis in 35.7% of cases, BPH in 35.7% of the cases, and P.Ca in 28.57% of the cases were observed. Thus, the nonmalignant cases constitute a majority. PSA, a marker specific for prostatic conditions, was significantly high in all the diseases compared to controls. A rise in serum PSA and PAP indicates prostatitis or, in combination with these two tests, decreased serum calcium shows advanced disease.

  15. Biomarkers of Prostatic Cancer: An Attempt to Categorize Patients into Prostatic Carcinoma, Benign Prostatic Hyperplasia, or Prostatitis Based on Serum Prostate Specific Antigen, Prostatic Acid Phosphatase, Calcium, and Phosphorus

    Sarwar, Shahana; Nyamath, Parveen; Ishaq, Mohammed


    Prostatitis, BPH, and P.Ca are the most frequent pathologies of the prostate gland that are responsible for morbidity in men. Raised levels of PSA are seen in different pathological conditions involving the prostate. PAP levels are altered in inflammatory or infectious or abnormal growth of the prostate tissue. Serum calcium and phosphorus levels were also found to be altered in prostate cancer and BPH. The present study was carried out to study the levels of PSA, PAP, calcium, and phosphorus in serum of patients with Prostatitis, BPH, or P.Ca and also to evaluate the relationship between them. Males in the age group of 50–85 years with LUTS disease symptoms and with PSA levels more than 4 ng/mL were included. A total of 114 patients were analyzed including 30 controls. Prostatitis in 35.7% of cases, BPH in 35.7% of the cases, and P.Ca in 28.57% of the cases were observed. Thus, the nonmalignant cases constitute a majority. PSA, a marker specific for prostatic conditions, was significantly high in all the diseases compared to controls. A rise in serum PSA and PAP indicates prostatitis or, in combination with these two tests, decreased serum calcium shows advanced disease. PMID:28168057

  16. Biomarkers of Prostatic Cancer: An Attempt to Categorize Patients into Prostatic Carcinoma, Benign Prostatic Hyperplasia, or Prostatitis Based on Serum Prostate Specific Antigen, Prostatic Acid Phosphatase, Calcium, and Phosphorus

    Shahana Sarwar


    Full Text Available Prostatitis, BPH, and P.Ca are the most frequent pathologies of the prostate gland that are responsible for morbidity in men. Raised levels of PSA are seen in different pathological conditions involving the prostate. PAP levels are altered in inflammatory or infectious or abnormal growth of the prostate tissue. Serum calcium and phosphorus levels were also found to be altered in prostate cancer and BPH. The present study was carried out to study the levels of PSA, PAP, calcium, and phosphorus in serum of patients with Prostatitis, BPH, or P.Ca and also to evaluate the relationship between them. Males in the age group of 50–85 years with LUTS disease symptoms and with PSA levels more than 4 ng/mL were included. A total of 114 patients were analyzed including 30 controls. Prostatitis in 35.7% of cases, BPH in 35.7% of the cases, and P.Ca in 28.57% of the cases were observed. Thus, the nonmalignant cases constitute a majority. PSA, a marker specific for prostatic conditions, was significantly high in all the diseases compared to controls. A rise in serum PSA and PAP indicates prostatitis or, in combination with these two tests, decreased serum calcium shows advanced disease.

  17. Relationships between serum osteoprotegerin, matrix metalloproteinase-2 levels and bone metabolism in postmenopausal women

    DAI Yi; SHEN Lin


    Background Serum osteoprotegerin (OPG) and matrix metalloproteinase-2 (MMP-2) have been shown to play a role in bone metabolism by degrading the bone matrix. The present study was undertaken to compare OPG and MMP-2 with bone mineral density and three markers (alkaline phosphatase (AKP), calcium and phosphorus) in postmenopausal women in Wuhan.Methods Serum OPG, MMP-2, and AKP of 78 Chinese postmenopausal women aged 48 to 65 were measured using enzyme-linked immunosorbent assay (ELISA). Bone mineral density was measured with dual energy X-ray absorptiometry (DEXA), and serum calcium and phosphorus were measured by auto biochemical analysis.Results Serum OPG and MMP-2 concentrations were significantly higher in postmenopausal women with osteoporosis ((127.6±6.3) ng/L; (1388±121) μg/L)) than those in age-matched normal controls ((72.3±2.4) ng/L; (1126±141) μg/L,P<0.01). Negative relationships were found between serum OPG, MMP-2 levels and bone mineral density in osteoporotic women. Adjusted by age and body mass index (BMI), the correlation of MMP-2 with bone mineral density of the neck of the femur disappeared. In osteoporotic women, negative correlations between OPG, MMP-2 levels and serum calcium were found (r=-0.216; r=-0.269, P<0.05), but positive correlations between OPG and serum AKP, serum phosphorus (r=0.235; r=0.124, P<0.05).Conclusions Significant correlations exist between serum OPG, MMP-2 levels and bone metabolism in high bone turnover of postmenopausal osteoporotic women. The concentrations of serum OPG and MMP-2 increase possibly as a concomitant event in the high bone turnover state, such as postmenopausal osteoporosis. Therefore serum OPG and MMP-2 could be used as indicators for the bone metabolism in postmenopausal osteoporotic women.

  18. Radioprotective effect of Panax ginseng on the phosphatases and lipid peroxidation level in testes of Swiss albino mice

    Kumar M.; Sharma M.K.; Saxena P.S.; Kumar A. [Rajasthan Univ., Jaipur (India)


    The Panax ginseng has been used as traditional medicine for past several years among oriental people. The present investigation has been made to assess the radioprotective efficacy of ginseng root extract in the testicular enzymes of Swiss albino mice. The Swiss albino mice were divided into different groups. Ginseng treated group: The animals were administered 10 mg/kg body weight ginseng root extract intraperitoneal (i.p.). Radiation treated group: The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 cm. Combination group: Animals were administered ginseng extract continuously for 4 d and on 4th day they were irradiated to 8 Gy gamma radiation after 30 min of extract administration. The animals from above groups were autopsied on day 1, 3, 7, 14 and 30. Biochemical estimations of acid and alkaline phosphatases and Lipid peroxidation (LPO) in testes were done. In ginseng treated group acid and alkaline phosphatases activity and LPO level did not show any significant alteration. In irradiated animals there was a significant increase in acid phosphatase activity and LPO level. However, significant decline in alkaline phosphatase activity was observed. The treatment of ginseng before irradiation causes significant decrease in acid phosphatase and LPO level and significant increase in alkaline phosphatase activity. One of the cause of radiation damage is lipid peroxidation. Due to lipid peroxidation, lysosomal membrane permeability alters and thus results in release of hydrolytic enzymes. So, an increase in acid phosphatase was noticed after radiation treatment. The alkaline phosphatase activity is associated with membrane permeability and different stages of spermatogenesis. Due to membrane damage and depletion of germ cells of testes after irradiation the enzyme activity was decreased. Ginseng markedly inhibits lipid peroxidation. It acts in indirect fashion to protect radical processes by inhibition of initiation of

  19. Restoration of parathyroid function after change of phosphate binder from calcium carbonate to lanthanum carbonate in hemodialysis patients with suppressed serum parathyroid hormone.

    Inaba, Masaaki; Okuno, Senji; Nagayama, Harumi; Yamada, Shinsuke; Ishimura, Eiji; Imanishi, Yasuo; Shoji, Shigeichi


    Control of phosphate is the most critical in the treatment of chronic kidney disease with mineral and bone disorder (CKD-MBD). Because calcium-containing phosphate binder to CKD patients is known to induce adynamic bone disease with ectopic calcification by increasing calcium load, we examined the effect of lanthanum carbonate (LaC), a non-calcium containing phosphate binder, to restore bone turnover in 27 hemodialysis patients with suppressed parathyroid function (serum intact parathyroid hormone [iPTH] ≦ 150 pg/mL). At the initiation of LaC administration, the dose of calcium-containing phosphate binder calcium carbonate (CaC) was withdrawn or reduced based on serum phosphate. After initiation of LaC administration, serum calcium and phosphate decreased significantly by 4 weeks, whereas whole PTH and iPTH increased. A significant and positive correlation between decreases of serum calcium, but not phosphate, with increases of whole PTH and iPTH, suggested that the decline in serum calcium with reduction of calcium load by LaC might increase parathyroid function. Serum bone resorption markers, such as serum tartrate-resistant acid phosphatase 5b, and N-telopeptide of type I collagen increased significantly by 4 weeks after LaC administration, which was followed by increases of serum bone formation markers including serum bone alkaline phosphatase, intact procollagen N-propeptide, and osteocalcin. Therefore, it was suggested that LaC attenuated CaC-induced suppression of parathyroid function and bone turnover by decreasing calcium load. In conclusion, replacement of CaC with LaC, either partially or totally, could increase parathyroid function and resultant bone turnover in hemodialysis patients with serum iPTH ≦ 150 pg/mL.

  20. Carbon and Nitrogen Sources Influence Tricalcium Phosphate Solubilization and Extracellular Phosphatase Activity by Talaromyces flavus.

    Stefanoni Rubio, P J; Godoy, M S; Della Mónica, I F; Pettinari, M J; Godeas, A M; Scervino, J M


    The aim of this work was to study phosphate (P) solubilization (and the processes involved in this event) by Talaromyces flavus (BAFC 3125) as a function of carbon and/or nitrogen sources. P solubilization was evaluated in NBRIP media supplemented with different carbon (glucose, sorbitol, sucrose, and fructose) and nitrogen (L-asparagine, urea, ammonium sulfate (AS), and ammonium nitrate (AN) combinations. The highest P solubilization was related to the highest organic acid production (especially gluconic acid) and pH drop for those treatments where glucose was present. Also P solubilization was higher when an inorganic nitrogen source was supplemented to the media when compared to an organic one. Although not being present an organic P source, phosphatase activity was observed. This shows that P mineralization and P solubilization can occur simultaneously, and that P mineralization is not induced by the enzyme substrate. The combination that showed highest P solubilization was for AN-glucose. The highest acid phosphatase activity was for AS-fructose, while for alkaline phosphatase were for AS-fructose and AN-fructose. Acid phosphatase activity was higher than alkaline. P solubilization and phosphatase activity (acid and alkaline) were influenced by the different carbon-nitrogen combinations. A better understanding of phosphate-solubilizing fungi could bring a better use of soil P.

  1. Phosphatase Activity of Microbial Populations in Different Milk Samples in Relation to Protein and Carbohydrate Content

    Sosanka Protim SANDILYA


    Full Text Available Cattle milk is a rich source of protein, carbohydrate, vitamins, minerals and all other major and micro nutrients. At a moderate pH, milk is an excellent media for the growth of microbes and thus, intake of raw milk is precarious. In this study, attempt was made for a qualitative study of eight raw milk samples of different varieties of cow and goat milk, collected from Jorhat district of Assam, India, on the basis of nutritional value and microbial population. The highest microbial population was found in the milk collected from cross hybrid variety of cow, whereas microbial contamination was the least in Jersey cow milk. Samples of C1 (Jersey cow variety showed presence of the highest amount of protein and carbohydrate content as compared to the others. Almost all the milk samples showed positive acid and alkaline phosphatase activity. Maximum acid phosphatase activity was observed in cross hybrid cow milk, whereas local cow milk exhibited the highest alkaline phosphatase activity. Phosphatase activity did not show any co-relationship with microbial population of the milk samples. Similarly, the protein and carbohydrate content of the samples did not have any significant impact on both acid and alkaline phosphatase activity.

  2. [Serum enzyme alterations during diabetic ketoacidosis. Prospective study of the behaviour of AP, AsAT, AIAT, and GGPT in 24 cases (author's transl)].

    Pérez-Jiménez, F; Muñoz, R; Jiménez Perepérez, J; Jiménez Alonso, J; Grilo, A; Benito, P


    Because of the scarcity of data on the changes of serum enzymes during diabetic ketoacidosis, the authors have prospectively studied the alterations of gamma-glutamyl transpeptidase (GGTP), alkaline phosphatase (AP), aspartate aminotransferase (AsAT), and alanine aminotransferase (AIAT) in this metabolic disturbance. The most significant finding was the frequent increase of AP activity on admission, with a systematic fall of the serum levels within the first 24 hours (p less than 0.001). This phenomenon was negatively correlated with patients' age (p less than 0.05), and the same occurred with the bone isoenzyme of AP (p less than 0.01). The remaining enzymes studies were always normal, thus suggesting that any increase of the serum levels of GGTP, AsAT, and AIAT found in a patient with diabetic ketoacidosis must arise the suspicion of an associated disturbance.

  3. Fabrication of titanium dioxide nanotube array and effects of its osteoblast proliferation and alkaline phosphatase activity%二氧化钛纳米管的制备及其对成骨细胞增殖和碱性磷酸酶活性的影响

    于卫强; 蒋欣泉; 张益琳; 张富强


    Objective To investigate preparation parameters of TiO_2 nanotube layer with anodization,and to evaluate the osteoblast activity on TiO_2 nanotube layer in vitro.Methods Titanium dioxide nanotube layer was grown using anodization method.TiO_2 nanotube layers with different structure were obtained by controlling the voltage and time of anodization and rinsing process after anodizatian.Pure titanium without anodization was used as control.Osteoblasts were cultivated on the anodizated TiO_2 nanotube,and the growth of osteoblasts was then evaluated and analyzed.Results The voltage,duration of anodization and rinsing process following anodization were key factors to affect morphology of TiO_2 nanotube layer.The area of osteoblast cultured on TiO_2 nanotube layer was larger than that on pure titanium.Results from methylthiazol tetrazolium(MTT) test showed that the proliferation of osteoblasts after 96 h cultivation on TiO_2 nanotube layer(0.62±0.02) was significantly higher than that on unanodizated titanium (0.55±0.03,P<0.05).Three weeks later,the alkaline phosphatase(ALP) activity on TiO_2 nanotube layer[(130.8±5.1) A405/mg]was significantly higher than that on unanodizated titanium[(109.6±4.5) A_(405)/mg,P<0.05].Conclusions The structure of TiO_2 nanotube layer was greatly associated with anedization voltage.The TiO_2 nanotube layer had a positive effect on osteoblast behavior.%目的 探讨阳极氧化过程中不同工艺条件对TiO_2纳米管形貌的影响,并对TiO_2纳米管对成骨细胞增殖和碱性磷酸酶相对活性的影响进行初步评定.方法 采用阳极氧化法在钛基底表面制备TiO_2纳米管,通过控制阳极氧化电压(5、15、20和25 V)、作用时间(1.5和3 h)以及阳极化后冲洗工艺(是否行超声处理),得到不同结构的TiO_2纳米管,扫描电镜观察TiO_2纳米管管径和管长.以未行阳极氧化的钛试件为对照组,以行阳极氧化(阳极氧化电压为15 V,作用时间为3 h,反应后行超

  4. Effect of two different doses of oral cholecalciferol supplementation on serum 25-hydroxy-vitamin D levels in healthy Indian postmenopausal women: A randomized controlled trial

    Niti Agarwal


    Full Text Available Aim: To compare the effect of two different doses (500 and 1000 IU/day of oral vitamin D3 (cholecalciferol on serum 25-hydroxy vitamin D [25(OHD] levels in apparently healthy postmenopausal Indian women. Materials and Methods: Serum 25(OHD, calcium with albumin, phosphorus, and alkaline phosphatase were measured in 92 apparently healthy postmenopausal women. The subjects were randomly assigned to one of the three groups and received supplementation for 3 months each. Each group received 1000 mg calcium carbonate daily while groups B and C received 500 and 1000 IU of cholecalciferol in addition, respectively. The tests were repeated after 3 months. Results: At baseline, 83.7% subjects had vitamin D deficiency (≤20 ng/mL. The difference in the percentage change in mean serum 25(OHD levels from baseline in group A (-30.5 ± 5.3%, group B (+8.9 ± 19.7%, and in group C (+97.8 ± 53.3% was statistically significant (P 20 ng/mL was achieved in 4.7% (1/21, 16% (4/25, and 66.67% (12/18 subjects in groups A, B, and C, respectively. No significant change was found in serum calcium, phosphorus, and alkaline phosphatase levels at 3 months in either of the groups from baseline. Conclusions: Standard dose of cholecalciferol available in "calcium tablets" (250 IU per 500 mg calcium carbonate is not adequate for achieving optimum serum 25(OHD levels in Indian postmenopausal women. Higher dose of vitamin D supplementation with 1000 IU/day (500 IU per 500 mg calcium carbonate daily is superior to the standard dose therapy. For achievement of optimum serum 25(OHD levels (>30 ng/mL in Indian postmenopausal women, still higher doses of vitamin D are likely to be required.

  5. The diagnostic value of alpha-fetoprotein, α-L-fucosidase,gamma-glutamyltransferase, alkaline phosphatase and carbohydrate antigen 19-9 in the diagnosis of primary hepatocellular carcinoma%甲胎蛋白、α-L-岩藻糖苷酶、γ-谷氨酰转移酶、碱性磷酸酶及糖类抗原19-9联合检测在原发性肝癌中的诊断价值



    Objective To investigate the diagnostic value of alpha-fetoprotein (AFP),α-L-fucosidase(AFU),gamma-glutamyltransferase(GGT),alkaline phosphatase (ALP) and carbohydrate antigen 19-9(CA19-9) in the diagnosis of primary hepatocellular carcinoma(PHC).Methods Seventy-eight PHC patients as PHC group,76 patients with other liver diseases as other liver diseases group and 60 healthy volunteers as control group.The level of serum AFP,AFU,GGT,ALP,CA19-9 and positive rate in each group were compared.Results The level of AFP,AFU,GGT,ALP,CA19-9 in PHC group were (276.3 ±72.6)μg/L,(63.1 ± 11.7) U/L,(268.5 ±57.2) U/L,(293.4 ±61.7) U/L and (56.9 ± 12.5) μg/L,in other liver diseases group were (81.6 ± 12.1) μg/L,(28.3 ±7.3) U/L,(54.9 ±7.8) U/L,(116.5 ± 23.8) U/L and (27.8 7.1) μ g/L,in control group were (13.8 ± 2.7) μ g/L,(12.6 ± 3.9) U/L,(12.3 ± 3.2) U/L,(47.2 ± 11.3) U/L and (12.9 ± 3.4) μ g/L.The level of above index in PHC group and other liver diseases group were significantly higher than those in control group,while the level of above index in PHC group were significantly higher than those in other liver diseases group (P < 0.01 or < 0.05).The positive rate of AFP,AFU,GGT,ALP,CA19-9 in PHC group were 62.8%(49/78),79.5%(62/78),83.3% (65/78),85.9% (67/78),66.7% (52/78),and significantly higher than those in other liver diseases group[31.6%(24/76),32.9%(25/76),27.6%(21/76),22.4%(17/76),15.8 %(12/76)].The positive rate of AFP in control group was 0,the positive rate of AFU,GGT,ALP,CA19-9 in control group were 1.7% (1/60),3.3% (2/60),1.7% (1/60),8.3% (5/60),which significantly lower than those in other two groups.The positive rate of combined detection in PHC group was 100.0%(78/78),which significantly higher than that in other liver diseases group [40.8%(31/76)] and control group[8.3%(5/60)](P< 0.01).Conclusion The combined detection of AFP,AFU,GGT,ALP,CA19-9 in PHC diagnosis has a higher sensitivity,and thus to make

  6. Anodes for alkaline electrolysis

    Soloveichik, Grigorii Lev


    A method of making an anode for alkaline electrolysis cells includes adsorption of precursor material on a carbonaceous material, conversion of the precursor material to hydroxide form and conversion of precursor material from hydroxide form to oxy-hydroxide form within the alkaline electrolysis cell.

  7. Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus

    Gilbert Christophe


    Full Text Available Abstract Background Reversible phosphorylation events within a polymerisation complex have been proposed to modulate capsular polysaccharide synthesis in Streptococcus pneumoniae. Similar phosphatase and kinase genes are present in the exopolysaccharide (EPS biosynthesis loci of numerous lactic acid bacteria genomes. Results The protein sequence deduced from the wzb gene in Lactobacillus rhamnosus ATCC 9595 reveals four motifs of the polymerase and histidinol phosphatase (PHP superfamily of prokaryotic O-phosphatases. Native and modified His-tag fusion Wzb proteins were purified from Escherichia coli cultures. Extracts showed phosphatase activity towards tyrosine-containing peptides. The purified fusion protein Wzb was active on p-nitrophenyl-phosphate (pNPP, with an optimal activity in presence of bovine serum albumin (BSA 1% at pH 7.3 and a temperature of 75°C. At 50°C, residual activity decreased to 10 %. Copper ions were essential for phosphatase activity, which was significantly increased by addition of cobalt. Mutated fusion Wzb proteins exhibited reduced phosphatase activity on p-nitrophenyl-phosphate. However, one variant (C6S showed close to 20% increase in phosphatase activity. Conclusion These characteristics reveal significant differences with the manganese-dependent CpsB protein tyrosine phosphatase described for Streptococcus pneumoniae as well as with the polysaccharide-related phosphatases of Gram negative bacteria.

  8. Structural Genomics of Protein Phosphatases

    Almo,S.; Bonanno, J.; Sauder, J.; Emtage, S.; Dilorenzo, T.; Malashkevich, V.; Wasserman, S.; Swaminathan, S.; Eswaramoorthy, S.; et al


    The New York SGX Research Center for Structural Genomics (NYSGXRC) of the NIGMS Protein Structure Initiative (PSI) has applied its high-throughput X-ray crystallographic structure determination platform to systematic studies of all human protein phosphatases and protein phosphatases from biomedically-relevant pathogens. To date, the NYSGXRC has determined structures of 21 distinct protein phosphatases: 14 from human, 2 from mouse, 2 from the pathogen Toxoplasma gondii, 1 from Trypanosoma brucei, the parasite responsible for African sleeping sickness, and 2 from the principal mosquito vector of malaria in Africa, Anopheles gambiae. These structures provide insights into both normal and pathophysiologic processes, including transcriptional regulation, regulation of major signaling pathways, neural development, and type 1 diabetes. In conjunction with the contributions of other international structural genomics consortia, these efforts promise to provide an unprecedented database and materials repository for structure-guided experimental and computational discovery of inhibitors for all classes of protein phosphatases.

  9. Effects of antiserums of cadherin, aminopeptidase N and alkaline phosphatase on the toxicities of Cry1Ac and Cry2Aa in Helicoverpa armigera (Lepidoptera: Noctuidae)%棉铃虫中肠钙粘蛋白、氨肽酶N及碱性磷酸酯酶的抗血清对Cry1Ac和Cry2Aa毒力的影响

    王冰洁; 袁向东; 赵曼; 刘臣; 陈琳; 梁革梅


    [Aim] Cry1A and Cry2A toxins play insecticidal roles by specifically binding with receptor proteins on insect midgut,and are widely used in Bt transgenic crops now.This study aims to further understand the action mechanisms of Cry2A and the roles of the functional receptor proteins of Cry1 A in the toxicity of Cry2A.[Methods] We firstly extracted brush border membrane vesicles (BBMV) of midgut,prepared antibody and antiserums of cadherin (CAD),aminopeptidase N (APN),and alkaline phosphatase (ALP) in Helicoverpa armigera.Then,after detecting the antiserums of these three receptor proteins in BBMV by Western blot,we compared the effects of antiserums of these three receptor proteins on the toxicities of Cry1Ac and Cry2Aa in the susceptible and Cry1Ac-resistant H.armigera (BtR) by the antibody blocking technology.[Results] For the susceptible H.armigera,the antiserums of these three receptor proteins not only significantly reduced the toxicity of Cry1 Ac,but also remarkably reduced the toxicity of Cry2Aa.Among them,anti-APN antiserum had the biggest impact on the toxicity of Cry1Ac,causing the mortality of H.armigera larvae to be reduced by 84.44%,and anti-ALP antiserum had the greatest effect on the toxicity of Cry2Aa,causing the larval mortality to be reduced by 71.04% compared with the control (without treatment by antiserum).The toxicity of Cry1Ac to Cry1Ac-resistant H.armigera (BtR) was obviously reduced and the toxicity of Cry2Aa also became less to these resistant larvae.The influences of the antiserums of these three receptors on the toxicity of Cry1Ac to the Cry1Acresistant strain of H.armigera (BtR) were smaller than those to the susceptible strain.Especially,the inhibition percentage of anti-CAD and-APN antiserums to the toxicity of Cry1 Ac decreased significantly.The effects of anti-CAD and-ALP antiserums on the toxicity of Cry2Aa to the Cry1Ac-resistant strain (BtR) and the susceptible strain were not significantly different,but the anti-APN antiserum

  10. Performance and Serum Hepatic Enzymes of Hy-Line W-36 Laying Hens Intoxicated with Dietary Carbon Tetrachloride

    Hadavi A


    Full Text Available An experiment was conducted to study the effects of carbon tetrachloride (CCl4 on post-peak performance and serum enzymes of Hy-Line W-36 laying hens from 32-36 weeks of age. The experiment was carried out with a total of 192 laying hens in a completely randomized block design. During the experiment laying hens were allocated to 4 groups consisted of T1 no CCl4 as control diet, T2, T3 and T4 control diet supplemented with 1, 3 and 5 mL CCl4/100 g diet, respectively. Each experimental group was divided into 6 blocks of 8 hens each. Egg production, cracked egg percentage and feed intake were recorded weekly. Blood samples were taken from wing veins of hens at the middle and end of the experiment to measure serum hepatic enzymes of alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase. Data showed that in comparison with the control group, the inclusion of CCl4 to the diets had no significant effect on performance parameters. However, by increasing the level of CCl4, egg production was linearly decreased and feed intake was linearly increased (P < 0.05. The effect of CCl4 on cracked eggs was significant and this effect was linearly increased (P < 0.05. Dietary supplementation of 3 and 5 mL CCl4 elevated the serum concentration of hepatic enzymes of alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase, linearly (P < 0.0001. In conclusion, the dietary supplementation of CCl4 has the ability to decrease the performance and egg quality. CCl4 is also a potent hepatic toxicity inducer and may damage liver hepatocytes. Therefore, the level of 3 mL CCl4 was assigned as the one had the maximum negative effect on serum hepatic enzymes concentration (maximum liver damage alongside the minimum negative effect on laying hen performance for further studies.

  11. Glucose-6-phosphatase deficiency

    Labrune Philippe


    Full Text Available Abstract Glucose-6-phosphatase deficiency (G6P deficiency, or glycogen storage disease type I (GSDI, is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, between the ages of 3 to 4 months by symptoms of hypoglycemia (tremors, seizures, cyanosis, apnea. Patients have poor tolerance to fasting, marked hepatomegaly, growth retardation (small stature and delayed puberty, generally improved by an appropriate diet, osteopenia and sometimes osteoporosis, full-cheeked round face, enlarged kydneys and platelet dysfunctions leading to frequent epistaxis. In addition, in GSDIb, neutropenia and neutrophil dysfunction are responsible for tendency towards infections, relapsing aphtous gingivostomatitis, and inflammatory bowel disease. Late complications are hepatic (adenomas with rare but possible transformation into hepatocarcinoma and renal (glomerular hyperfiltration leading to proteinuria and sometimes to renal insufficiency. GSDI is caused by a dysfunction in the G6P system, a key step in the regulation of glycemia. The deficit concerns the catalytic subunit G6P-alpha (type Ia which is restricted to expression in the liver, kidney and intestine, or the ubiquitously expressed G6P transporter (type Ib. Mutations in the genes G6PC (17q21 and SLC37A4 (11q23 respectively cause GSDIa and Ib. Many mutations have been identified in both genes,. Transmission is autosomal recessive. Diagnosis is based on clinical presentation, on abnormal basal values and absence of hyperglycemic response to glucagon. It can be confirmed by demonstrating a deficient activity of a G6P system component in a liver biopsy. To date, the diagnosis is most

  12. Glucose-6-phosphatase deficiency.

    Froissart, Roseline; Piraud, Monique; Boudjemline, Alix Mollet; Vianey-Saban, Christine; Petit, François; Hubert-Buron, Aurélie; Eberschweiler, Pascale Trioche; Gajdos, Vincent; Labrune, Philippe


    Glucose-6-phosphatase deficiency (G6P deficiency), or glycogen storage disease type I (GSDI), is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, between the ages of 3 to 4 months by symptoms of hypoglycemia (tremors, seizures, cyanosis, apnea). Patients have poor tolerance to fasting, marked hepatomegaly, growth retardation (small stature and delayed puberty), generally improved by an appropriate diet, osteopenia and sometimes osteoporosis, full-cheeked round face, enlarged kydneys and platelet dysfunctions leading to frequent epistaxis. In addition, in GSDIb, neutropenia and neutrophil dysfunction are responsible for tendency towards infections, relapsing aphtous gingivostomatitis, and inflammatory bowel disease. Late complications are hepatic (adenomas with rare but possible transformation into hepatocarcinoma) and renal (glomerular hyperfiltration leading to proteinuria and sometimes to renal insufficiency). GSDI is caused by a dysfunction in the G6P system, a key step in the regulation of glycemia. The deficit concerns the catalytic subunit G6P-alpha (type Ia) which is restricted to expression in the liver, kidney and intestine, or the ubiquitously expressed G6P transporter (type Ib). Mutations in the genes G6PC (17q21) and SLC37A4 (11q23) respectively cause GSDIa and Ib. Many mutations have been identified in both genes,. Transmission is autosomal recessive. Diagnosis is based on clinical presentation, on abnormal basal values and absence of hyperglycemic response to glucagon. It can be confirmed by demonstrating a deficient activity of a G6P system component in a liver biopsy. To date, the diagnosis is most commonly confirmed

  13. A critical evaluation of a specific radioimmunoassay for prostatic acid phosphatase

    Goldenberg, S.L.; Silver, H.K.; Sullivan, L.D.; Morse, M.J.; Archibald, E.L.


    A radioimmunoassay (RIA) method for acid phosphatase detection was compared to a standard enzyme assay using sera from 210 normal volunteers and 285 patients with prostatic disease. Statistical and clinical comparisons were made between defined subgroups. All 55 normal females had RIA detectable serum acid phosphatase, implying that this assay cannot be entirely specific for enzyme of prostatic origin. Urinary catheterization did not affect acid phosphatase levels. In all stages of carcinoma there were more acid phosphatase elevations by the RIA method than enzyme method, but neither assay could differentiate intercapsular cancer from benign prostatic hyperplasia. A small number of patients with biopsy proven negative nodules had marginally elevated values, suggesting an obligation for closer follow-up. The RIA method may be superior for monitoring patients with more advanced malignancy. Additional practical advantages of the RIA include relative simplicity and elimination of the special serum handling required for the enzyme assay.

  14. Alkaline battery operational methodology

    Sholklapper, Tal; Gallaway, Joshua; Steingart, Daniel; Ingale, Nilesh; Nyce, Michael


    Methods of using specific operational charge and discharge parameters to extend the life of alkaline batteries are disclosed. The methods can be used with any commercial primary or secondary alkaline battery, as well as with newer alkaline battery designs, including batteries with flowing electrolyte. The methods include cycling batteries within a narrow operating voltage window, with minimum and maximum cut-off voltages that are set based on battery characteristics and environmental conditions. The narrow voltage window decreases available capacity but allows the batteries to be cycled for hundreds or thousands of times.

  15. A contribution for the definition of serum chemistry values in captive adults Antillean manatees (Trichechus manatus manatus Linnaeus, 1758).

    Silva, F M O; Vergara-Parente, J E; Gomes, J K N; Teixeira, M N; Lima, R P


    Serum chemistry analyses represents a fundamental tool for the diagnosis and understanding of diseases in marine mammals. Although several studies are being conducted within the field of clinical pathology, haematological and serum chemistry data for Antillean manatees are deficient. The purpose of this study was to determine serum chemistry values for captive Antillean manatees within the CMA/Ibama facility in Brazil. Serum samples were obtained from five captive adult Antillean manatees fed with seagrass and analysed for aspartate aminotransferase, alanine aminotransferase, bilirubin, alkaline phosphatase, urea, creatinine, glucose, triglycerides, cholesterol, total protein, albumin, globulin, phosphate, chloride, calcium and uric acid. Blood chemistry parameters were determined using a semi-automatic analyzer. Maximum, minimum, mean and standard deviations were calculated for each serum chemistry parameter. Differences on the values of males and females were verified using an unpaired Student's t-test. All the parameters analysed were similar between sexes, with exception of AP, which was higher in females (191.43 +/- 31.86 U/l). Alanine aminotransferase and uric acid values for Trichechus manatus manatus are reported for the first time in this paper. This study is the first to report serum chemistry parameter values for long-term captive male and female Antillean manatees. Therefore, the lower values of albumin, phosphate, chloride, cholesterol and triglycerides obtained here highlight the importance of clinical pathology during health monitoring of captive marine mammals.

  16. Uranium in alkaline rocks

    Murphy, M.; Wollenberg, H.; Strisower, B.; Bowman, H.; Flexser, S.; Carmichael, I.


    Geologic and geochemical criteria were developed for the occurrence of economic uranium deposits in alkaline igneous rocks. A literature search, a limited chemical analytical program, and visits to three prominent alkaline-rock localities (Ilimaussaq, Greenland; Pocos de Caldas, Brazil; and Powderhorn, Colorado) were made to establish criteria to determine if a site had some uranium resource potential. From the literature, four alkaline-intrusive occurrences of differing character were identified as type-localities for uranium mineralization, and the important aspects of these localities were described. These characteristics were used to categorize and evaluate U.S. occurrences. The literature search disclosed 69 U.S. sites, encompassing nepheline syenite, alkaline granite, and carbonatite. It was possible to compare two-thirds of these sites to the type localities. A ranking system identified ten of the sites as most likely to have uranium resource potential.

  17. Clinical retrospective analysis of the relationship between serum calcium levels and apathetic hyperthyroidism%淡漠型甲状腺功能亢进症与血钙水平关系的临床回顾分析

    吴伟华; 孙振杰; 孙丽芳; 匡洪宇


    分析典型甲状腺功能亢进症(甲亢)与淡漠型甲亢患者血钙水平的差异,发现淡漠型甲亢血钙较典型甲亢显著增高,血清骨源性碱性磷酸酶(BAP)则显著降低(均P<0.01).淡漠型甲亢临床表现不典型的原因可能与血钙过度增高有关.%The difference in serum calcium levels between patients with typical hyperthyroidism and apathetic hyperthyroidism was analyzed. Serum calcium levels in patients with apathetic hyperthyroidism were higher than those in patients with typical hyperthyroidism, while bone-specific alkaline phosphatase levels were lower. The atypical manifestation of apathetic hyperthyroidism may be due to the significant elevation of serum calcium level.

  18. Phosphate-solubility and phosphatase activity in Gangetic alluvial soil as influenced by organophosphate insecticide residues.

    Majumder, Shyam Prasad; Das, Amal Chandra


    An experiment was conducted under laboratory conditions to investigate the effect of four organophosphate insecticides, viz. monocrotophos, profenophos, quinalphos and triazophos at their field application rates (0.75, 1.0, 0.5 and 0.6 kg a.i.ha(-1), respectively), on the growth and activities of phosphate solubilizing microorganisms in relation to availability of insoluble phosphates in the Gangetic alluvial soil of West Bengal, India. The proliferation of phosphate solubilizing microorganisms was highly induced with profenophos (38.3%), while monocrotophos exerted maximum stimulation (20.8%) towards the solubility of insoluble phosphates in soil. The phosphatase activities of the soil (both acid phosphatase and alkaline phosphatase) were significantly increased due to the incorporation of the insecticides in general, and the augmentation was more pronounced with quinalphos (43.1%) followed by profenophos (27.6%) for acid phosphatase, and with monocrotophos (25.2%) followed by profenophos (16.1%) for alkaline phosphatase activity in soil. The total phosphorus was highly retained by triazophos (19.9%) followed by monocrotophos (16.5%), while incorporation of triazophos and quinalphos manifested greater availability of water soluble phosphorus in soil.

  19. Proliferation,apoptosis and expression of tissue non-specific alkaline phosphatase in cells of stratum intermedium during amelogenesis in postnatal stage of BALB/c mouse%小鼠釉质发育过程中成釉器中间层细胞增殖、凋亡以及组织非特异性碱性磷酸酶的表达

    唐开亮; 李纾; 肖长杰; 王建; 于兰; 于西佼


    目的:观察小鼠釉质发育过程中成釉器中间层细胞的增殖、凋亡和其组织非特异性碱性磷酸酶(tissue nonspecific alkaline phosphatase,TNSALP)的表达,探讨中间层在釉质形成过程中的可能作用.方法:取出生后1、3、5、7、9、11、15d的BALB/c小鼠56只,解剖分离含下颌第一磨牙区的下颌骨,脱钙,制片,HE染色进行牙釉质发育情况的组织学观察,分别采用原位末端标记法(TUNEL法)、SP免疫组织化学技术、PV二步法观察中间层细胞的凋亡及检测Bcl-2、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),并对TNSALP进行组织学定位.通过Image-pro plus 6.0图像分析软件.对图像进行半定量分析.采用SPSS13.0软件包进行t检验、单因素方差分析及SNK-q检验.结果:出生后1d.中间层细胞增殖细胞核抗原由比成釉细胞表达高的强阳性表达逐渐降低;至7d时,其表达为阴性,而且中间层细胞随釉质的发育逐渐凋亡.中间层细胞先于成釉细胞表达TNSALP,出生后5d时即呈现强阳性染色,而后其表达又逐步减弱;成釉细胞7d时开始出现阳性,且呈逐渐增强趋势.结论:中间层细胞的增殖和凋亡及其TNSALP的表达与成釉细胞具有相关性,提示中间层细胞有可能参与成釉细胞的分化及釉质形成.

  20. Milk-derived proteins and minerals alter serum osteocalcin in prepubertal boys after 7 days

    Mark, A.B.; Hoppe, Camilla; Michaelsen, K.F.


    We have previously shown that at equal protein content, milk, but not meat, decreased bone turnover in boys. This suggested that milk-derived components are important for bone metabolism. In the present study, we hypothesized that milk-derived proteins (whey and casein) affect bone turnover during...... growth depending on the content of milk minerals (calcium and phosphorus). This was a randomized, parallel, double-blind study. Eight-year-old boys (n = 57) received 1 of 4 milk drinks: whey protein with low or high content of minerals, or casein protein with low or high content of minerals. The amount...... of whey and casein was identical to their content in 1.5 L of milk. We measured serum osteocalcin (sOC), bone-specific alkaline phosphatase, and C-terminal telopeptides of type 1 collagen (immunoassay) and estimated dietary intake (3-day weighed food record) at baseline and after 7 days. Only s...

  1. The effects of hydroalcoholic extract of Portulaca Oleracea on the serum concentreation of Hepatic enzymes in Rats

    Ali Zarei


    Full Text Available Background Hyperlipidemia can be cause a variety of diseases such as atherosclerosis, diabetes and fatty liver and subsequent liver enzyme increases. The Portulaca Oleracea plant has hypoglycemic and hypolipidemic properties. Therefore, in this study the effect of Portulca Oleracea herb extract on serum liver enzymes including aspartate aminotransferase (SGOT or AST, alanine aminotransferase (SGPT or ALT and alkaline phosphatase (ALP in rats were studied. Materials and Methods: In this experimental study, 60 Wistar rats were divided into 6 groups (n=10. Control group with normal diet, fat diet group and other groups, the experimental group received the same diet plus fat Portulaca oleracea extract maximum dose (800, the mean dose of (400, and a minimum dose of (200 mg / kg or intraperitoneally injection (ip and sort of Atorvastatin (10 mg kg. After the end of this period (21 days, blood sampling was performed and collected data were analyzed using the t and Tukey test, and SPSS software version 11.5. Results Comparison of statistical results indicated that alanine aminotransferase (ALT and Alkaline phosphatase (ALP increase in the control group that received only fatty foods, while the experimental groups received extract of Portulaca Oleracea, and groups receiving Atorvastatin had reduced levels of liver enzymes. Conclusion: Regarding hypoglycemic and hypolipidemic antioxidant activity of the extract and its effect on reducing liver enzymes, plant extracts can be recommended to improve liver function.

  2. The role of phosphatases in the initiation of skeletal mineralization.

    Millán, José Luis


    Endochondral ossification is a carefully orchestrated process mediated by promoters and inhibitors of mineralization. Phosphatases are implicated, but their identities and functions remain unclear. Mutations in the tissue-nonspecific alkaline phosphatase (TNAP) gene cause hypophosphatasia, a heritable form of rickets and osteomalacia, caused by an arrest in the propagation of hydroxyapatite (HA) crystals onto the collagenous extracellular matrix due to accumulation of extracellular inorganic pyrophosphate (PPi), a physiological TNAP substrate and a potent calcification inhibitor. However, TNAP knockout (Alpl(-/-)) mice are born with a mineralized skeleton and have HA crystals in their chondrocyte- and osteoblast-derived matrix vesicles (MVs). We have shown that PHOSPHO1, a soluble phosphatase with specificity for two molecules present in MVs, phosphoethanolamine and phosphocholine, is responsible for initiating HA crystal formation inside MVs and that PHOSPHO1 and TNAP have nonredundant functional roles during endochondral ossification. Double ablation of PHOSPHO1 and TNAP function leads to the complete absence of skeletal mineralization and perinatal lethality, despite normal systemic phosphate and calcium levels. This strongly suggests that the Pi needed for initiation of MV-mediated mineralization is produced locally in the perivesicular space. As both TNAP and nucleoside pyrophosphohydrolase-1 (NPP1) behave as potent ATPases and pyrophosphatases in the MV compartment, our current model of the mechanisms of skeletal mineralization implicate intravesicular PHOSPHO1 function and Pi influx into MVs in the initiation of mineralization and the functions of TNAP and NPP1 in the extravesicular progression of mineralization.

  3. Serum 5'nucleotidase activity in rats: a method for automated analysis and criteria for interpretation.

    Carakostas, Michael C.; Power, Richard J.; Banerjee, Asit K.


    A manual kit for determining serum 5'nucleotidase (5'NT, EC activity was adapted for use with rat samples on a large discrete clinical chemistry analyzer. The precision of the method was good (within-run C.V. = 2.14%; between-run C.V. = 5.5%). A comparison of the new automated method with a manual and semi-automated method gave regression statistics of y = 1.18X -3.66 (Sy. x = 4.54), and y = 0.733X + 1.97 (Sy. x = 1.69), respectively. Temperature conversion factors provided by the kit manufacturer for human samples were determined to be inaccurate for converting results from rat samples. Analysis of components contributing to normal variation in rat serum 5'NT activity showed age and sex to be major factors. Increased serum 5'NT activity was observed in female rats when compared to male rats beginning at about 5 to 6 weeks of age. An analysis of variance of serum 5'NT, alkaline phosphatase, and GGT activities observed over a 9-week period in normal rats suggests several advantages for 5'NT as a predictor of biliary lesions in rats.

  4. Alteration in serum and bone component findings induced in streptozotocin-diabetic rats is restored by zinc acexamate.

    Uchiyama, Satoshi; Yamaguchi, Masayoshi


    The effect of zinc acexamate in streptozotocin (STZ)-induced diabetic rats was investigated. Rats received a single subcutaneous administration of STZ (6.0 mg/100 g body weight), and the animals were orally administered once daily for 14 days with zinc acexamate (2.5, 5 or 10 mg/100 g body weight). The administration of STZ caused a significant increase in serum glucose, triglyceride and calcium levels and a significant decrease in body weight, serum zinc and inorganic phosphorus levels, indicating diabetic condition. Moreover, calcium content, alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the femoral-diaphyseal and -metaphyseal tissues were significantly reduced in STZ-diabetic rats. The change in these serum and bone components of STZ-diabetic rats was significantly restored by the oral administration of zinc acexamate (2.5, 5 or 10 mg Zn/100 g body weight). The restoration of bone components was not seen by the oral administration of zinc sulfate (2.5 mg Zn/100 g) for 14 days. Moreover, when the femoral-diaphyseal and -metaphyseal tissues obtained at 14 days after STZ administration were cultured for 48 h in a medium containing either vehicle or zinc acexamate (10(-5) M), the femoral calcium content and alkaline phosphatase activity were significantly increased in vitro. The effect of zinc acexamate was completely abolished in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis. The present study demonstrates that the oral administration of zinc acexamate has a preventive effect on STZ-induced diabetic condition in rats, and that it can restorate bone loss of STZ-induced diabetes in vivo.

  5. Biomarkers for the activation of calcium metabolism in dairy cows: elevation of tartrate-resistant acid phosphatase activity by lowering dietary cation-anion difference is associated with the prevention of milk fever.

    Kurosaki, Naotoshi; Yamato, Osamu; Sato, Jun; Naito, Yoshihisa; Mori, Fuminobu; Imoto, Seiichi; Maede, Yoshimitsu


    In our previous study, it was demonstrated that the administration of anion salts, which slightly lower the dietary cation-anion difference (DCAD), in the prepartum period is safe and effective for preventing milk fever in multiparous cows. In the present study, several biomarkers, which might show activation of Ca metabolism, were analyzed using stored samples in the previous study to investigate the mechanism of the preventive effect on milk fever by lowering DCAD. Changes in bone-specific alkaline phosphatase activity, osteocalcin and insulin-like growth factor I concentrations in serum were almost the same among the three groups of multiparous cows with or without the oral administration of anion salts, while the levels of these serum biomarkers in the group of primiparous cows (heifer group) were much higher compared with those in the three multiparous groups throughout the experimental period. Urinary deoxypyridinoline excretion was not a useful biomarker for dairy cows because it hardly changed during the peripartum period in all groups. However, serum tartrate-resistant acid phosphatase (TRAP) activity, which is known as a biomarker of osteoclast activity, was well associated with the administration of anion salts lowering DCAD because among the three multiparous groups, only the group of multiparous cows fed the anion salts (anion group) showed an increased level, which rose to the level in the heifer group, and was markedly higher than those in the other control groups of multiparous cows. The increased activity of serum TRAP in the anion group suggested that Ca in the plasma pool was mobilized smoothly from bone-bound Ca via mature osteoclasts at parturition, which might be due to prior activation under mild acidosis induced by slightly lowering DCAD. Therefore, TRAP was the best biomarker to monitor the activation of Ca metabolism in dairy cows fed anion salts.

  6. Alkaline earth metal thioindates

    Ivanov-Ehmin, B.N.; Ivlieva, V.I.; Filatenko, L.A.; Zajtsev, B.E.; Kaziev, G.Z.; Sarabiya, M.G.


    Alkaline earth metal thioindates of MIn/sub 2/S/sub 4/ composition were synthesized by interaction of alkaline earth metal oxoindates with hydrogen sulfide during heating. Investigation into the compounds by X-ray analysis showed that calcium compound crystallizes in cubic crystal system and strontium and barium compounds in rhombic crystal system. Lattice parameters and the number of formula units were determined. Thioindates of M/sub 3/In/sub 2/S/sub 6/ composition were synthesized, their individuality was shown.

  7. Serum fluoride and sialic acid levels in osteosarcoma.

    Sandhu, R; Lal, H; Kundu, Z S; Kharb, S


    Osteosarcoma is a rare malignant bone tumor most commonly occurring in children and young adults presenting with painful swelling. Various etiological factors for osteosarcoma are ionizing radiation, family history of bone disorders and cancer, chemicals (fluoride, beryllium, and vinyl chloride), and viruses. Status of fluoride levels in serum of osteosarcoma is still not clear. Recent reports have indicated that there is a link between fluoride exposure and osteosarcoma. Glycoproteins and glycosaminoglycans are an integral part of bone and prolonged exposure to fluoride for long duration has been shown to cause degradation of collagen and ground substance in bones. The present study was planned to analyze serum fluoride, sialic acid, calcium, phosphorus, and alkaline phosphatase levels in 25 patients of osteosarcoma and age- and sex-matched subjects with bone-forming tumours other than osteosarcoma and musculo-skeletal pain (controls, 25 each). Fluoride levels were analyzed by ISE and sialic acid was analyzed by Warren's method. Mean serum fluoride concentration was found to be significantly higher in patients with osteosarcoma as compared to the other two groups. The mean value of flouride in patients with other bone-forming tumors was approximately 50% of the group of osteosarcoma; however, it was significantly higher when compared with patients of group I. Serum sialic acid concentration was found to be significantly raised in patients with osteosarcoma as well as in the group with other bone-forming tumors as compared to the group of controls. There was, however, no significant difference in the group of patients of osteosarcoma when compared with group of patients with other bone-forming tumors. These results showing higher level of fluoride with osteosarcoma compared to others suggesting a role of fluoride in the disease.

  8. Effects of synthetic detergents on in vivo activity of tissue phosphatases and succinic dehydrogenase from Mystus vittatus.

    Mohan, D; Verma, S R


    African catfish (Mystus vittatus) were exposed to three sub-lethal concentrations of Swascofix E45 (13.8, 9.2 and 4.6 mg/l) and Swascol 3L (69.3, 46.2 and 23.1 mg/l) for 15 and 30 days, and their effects on alkaline and acid phosphatase, and succinic dehydrogenase in liver, kidney and intestine were measured. The enzymes were found to be inhibited in all the tissues. Maximum inhibition (38.44%) was observed in liver alkaline phosphatase activity after 30 days with the highest concentration of Swascofix E45 and the lowest inhibition (0.118%) was found in kidney acid phosphatase activity with the lowest concentration of Swascol 3L after 15 days. Insignificant enzyme stimulation in some cases was also observed.

  9. Effects of synthetic detergents on in vivo activity of tissue phosphatases and succinic dehydrogenase from Mystus vittatus

    Mohan, D.; Verma, S.R.


    African catfish (Mystus vittatus) were exposed to three sub-lethal concentrations of Swascofix E45 (13.8, 9.2 and 4.6 mg/l) and Swascol 3L (69.3, 46.2 and 23.1 mg/l) for 15 and 30 days, and their effects on alkaline and acid phosphatase, and succinic dehydrogenase in liver, kidney and intestine were measured. The enzymes were found to be inhibited in all the tissues. Maximum inhibition (38.44%) was observed in liver alkaline phosphatase activity after 30 days with the highest concentration of Swascofix E45 and the lowest inhibition (0.118%) was found in kidney acid phosphatase activity with the lowest concentration of Swascol 3L after 15 days. Insignificant enzyme stimulation in some cases was also observed.

  10. The Increased Level of Serum p53 in Hepatitis B-Associated Liver Cirrhosis

    Parisa Shahnazari


    Full Text Available Background: The ability of tumour suppressor protein p53 (P53 to regulate cell cycle processes can be modulated by hepatitis B virus (HBV. While preliminary evidences indicates the involvement of protein-x of HBV (HBx in altering p53 DNA binding, no further data have been accumulated for the significance of serum p53 in chronic hepatitis B virus infected patients. Methods: 72 non-cirrhotic and 19 cirrhotic patients infected by HBV were enrolled for the analysis in this study. Enzyme linked immunosorbent assay (ELISA was performed to study the concentrations of serum p53 protein. The tertiary structures of HBx and P53 were docked by Z-dock and Hex servers for in-silico protein-protein interaction analysis. Results: There was a significant association between the serum p53 and cirrhosis (OR=1.81 95% CI: 1.017-3.2, P=0.044. Cirrhotic patients had higher level of serum p53 compare with chronic infection of HBV (1.98±1.22 vs. 1.29±0.72 U/ml, P=0.05. No evidence of correlation was seen between the different variables such as age, gender, log viral load, serum alkaline phosphatase (ALP and alanine aminotransferase (ALT with serum p53. Tertiary model shows that the amino acid residues from Arg110 to Lys132 of the N-terminal of P53 which is critical for ubiquitination, are bonded to a region in N- terminal of HBx amino acid residues from Arg19 to Ser33. Conclusion: There is an increase in serum p53 in HBV-related cirrhosis patients. In this case, HBx might be responsible for such higher concentration of p53 through HBx-p53 protein-protein interaction, as is shown by molecular modeling approach.

  11. Phage & phosphatase: a novel phage-based probe for rapid, multi-platform detection of bacteria.

    Alcaine, S D; Pacitto, D; Sela, D A; Nugen, S R


    Genetic engineering of bacteriophages allows for the development of rapid, highly specific, and easily manufactured probes for the detection of bacterial pathogens. A challenge for novel probes is the ease of their adoption in real world laboratories. We have engineered the bacteriophage T7, which targets Escherichia coli, to carry the alkaline phosphatase gene, phoA. This inclusion results in phoA overexpression following phage infection of E. coli. Alkaline phosphatase is commonly used in a wide range of diagnostics, and thus a signal produced by our phage-based probe could be detected using common laboratory equipment. Our work demonstrates the successful: (i) modification of T7 phage to carry phoA; (ii) overexpression of alkaline phosphatase in E. coli; and (iii) detection of this T7-induced alkaline phosphatase activity using commercially available colorimetric and chemilumiscent methods. Furthermore, we demonstrate the application of our phage-based probe to rapidly detect low levels of bacteria and discern the antibiotic resistance of E. coli isolates. Using our bioengineered phage-based probe we were able to detect 10(3) CFU per mL of E. coli in 6 hours using a chemiluminescent substrate and 10(4) CFU per mL within 7.5 hours using a colorimetric substrate. We also show the application of this phage-based probe for antibiotic resistance testing. We were able to determine whether an E. coli isolate was resistant to ampicillin within 4.5 hours using chemiluminescent substrate and within 6 hours using a colorimetric substrate. This phage-based scheme could be readily adopted in labs without significant capital investments and can be translated to other phage-bacteria pairs for further detection.

  12. Growth and extracellular phosphatase activity of arbuscular mycorrhizal hyphae as influenced by soil organic matter

    Joner, E.J.; Jakobsen, I.


    Two experiments were set up to investigate the influence of soil organic matter on growth of arbuscular mycorrhizal (AM) hyphae and concurrent changes in soil inorganic P, organic P and phosphatase activity. A sandy loam soil was kept for 14 months under two regimes (outdoor where surplus...... differing in organic matter were placed in six parallel hyphal compartments (HC) separated from the RC with a 37 mu m mesh. In the first experiment, using Glomus caledonium, hyphal length densities were measured in the HC after 31 days. Added straw increased hyphal length densities by 34 and 62% for soil...... length density was twice as high in soil with added straw compared to the two other treatments. Mycorrhizal colonization resulted in lower activity of acid phosphatase in the HC for two out of three treatments. Alkaline phosphatase activity was only decreased by mycorrhiza in soil without organic matter...

  13. Correlation of Serum Leptin Level with Bone Mineral Density and Bone Turnover Markers in Chinese Adolescent Dancers



    Objective To investigate plasma leptin concentrations in adolescent female dancers and to determine whether leptin has some effects on their bone mineral density (BMD) and bone turnover markers. Methods Sixty dancers aged 15-17 years and 77 healthy controls were enrolled in the study. Bone mineral density (BMD) and body composition were detected by dual energy X-ray absorptiometry. Serum leptin concentrations were measured by radioimmunoassay (RIA). Two bone turnover markers, bone-specific alkaline phosphatase (BAP) and tartrate-resistant acid phosphatase(TRACP), were determined by ELISA. Results The dancers had a lower fat mass and a lower leptin level than the controls, while they had a relatively higher BMD of the total body and legs after adjustment for BMI and age. The levels of bone resorption and formation of markers were higher in the dancers than in the controls. Leptin was positively correlated with BMI, body weight, fat mass, and percentage of body fat. In dancers, Leptin was positively correlated with the BMD of the total body and the left leg. However, after adjustment for BMI, no correlation of serum leptin concentrations with BMD values was found in either dancers or controls. Nor correlation was found between leptin and bone turnover markers after adjustment for BMI. Conclusion The leptin profile is different between the controls and the dancers with a lower BMI and a lower fat mass. Circulating plasma leptin level depends on BMI and is not a direct determinant of BMD in Chinese adolescent dancers.

  14. Prostatic acid phosphatase by radioimmunoassay

    Lindholm, G.R.; Stirton, M.S.; Liedtke, R.J.; Batjer, J.D.


    Prostatic acid phosphatase values in 98 patients with prostatic carcinoma were measured by a commmercial radioimmunoassay (RIA) and by enzymatic assay. Forty-three carcinomas were staged by rigorous pathological criteria. Patients (N = 129) with benign prostatic hyperplasia were the control group. At 94% specificity, sensitivities of the RIA vs the enzymatic assay for clinically staged patients were as follows: stage A, 22% vs 6%; B, 29% vs 10%; C, 52% vs 38%; and D, 87% vs 80%. However, none of the seven patients with pathological stage A and B disease had a positive test result, and we suggest that variability in staging criteria accounts for the discrepant sensitivity claims reported. Prostatic acid phosphatase RIA should not be used for screening but as an adjunct for staging known prostatic carcinoma.

  15. Short-term bisphosphonate treatment reduces serum 25(OH) vitamin D3 and alters values of parathyroid hormone, pentosidine, and bone metabolic markers

    Kamimura, Mikio; Uchiyama, Shigeharu; Nakamura, Yukio; Ikegami, Shota; Mukaiyama, Keijiro; Kato, Hiroyuki


    This study aimed to clarify the effects of short-term bisphosphonate (BP) administration in Japanese osteoporotic patients retrospectively. Daily minodronate (MIN) at 1 mg/day (MIN group) or weekly risedronate (RIS) at 17.5 mg/week (RIS group) was primarily prescribed for each patient. We analyzed the laboratory data of 35 cases (18 of MIN and 17 of RIS) before the start of treatment and at 4 months afterward. The changes in 25(OH)D3, whole parathyroid hormone (PTH), serum pentosidine, and the bone turnover markers urinary cross-linked N-telopeptide of type I collagen (NTX), serum tartrate-resistant acid phosphatase (TRACP)-5b, bone-specific alkaline phosphatase (BAP), and undercarboxylated osteocalcin were evaluated. Overall, serum 25(OH)D3 was significantly decreased from 21.8 to 18.4 ng/mL at 4 months, with a percent change of −14.7%. Whole PTH increased significantly from 23.4 to 30.0 pg/mL, with a percent change of 32.1%. Serum pentosidine rose from 0.0306 to 0.0337 μg/mL, with a percent change of 15.2%. In group comparisons, 25(OH)D3 and pentosidine showed comparable changes in both groups after 4 months of treatment, whereas whole PTH became significantly more increased in the MIN group. All bone turnover markers were significantly decreased at 4 months in both groups. Compared with the RIS group, the MIN group exhibited significantly larger value changes for urinary NTX, serum TRACP-5b, and BAP at the study end point. This study demonstrated that serum 25(OH)D3 became significantly decreased after only 4 months of BP treatment in Japanese osteoporotic patients and confirmed that MIN more strongly inhibited bone turnover as compared with RIS.

  16. The diagnostic and prognostic value of serum bone Gla protein (osteocalcin) in patients with recurrent breast cancer

    Kamby, C; Egsmose, C; Söletormos, G


    bone survey. The sites of recurrence were bone (61%), bone marrow (46%), soft tissue (52%), lung (13%), pleura (11%), liver (4%), and brain (2%). Radiology and bone biopsy served as key diagnoses as to the presence or absence of bone metastases. The diagnostic efficiency of B-scan and S-AP was greater...... than that of S-BGP, and the result of BGP measurement was associated with neither extent nor number of bone metastases. However, the BGP values were significantly lower in patients who had visceral metastases, and the median duration of survival after recurrence was 13 months for patients with low S......Serum bone Gla protein (S-BGP), a marker of bone metabolism, was measured in 60 patients included in a staging programme for recurrent breast cancer. Other diagnostic procedures comprised S-alkaline phosphatase (S-AP), bone scan (B-scan), bilateral iliac crest bone marrow biopsies, and radiological...

  17. Alkaline broadening in Stars

    De Kertanguy, A


    Giving new insight for line broadening theory for atoms with more structure than hydrogen in most stars. Using symbolic software to build precise wave functions corrected for ds;dp quantum defects. The profiles obtained with that approach, have peculiar trends, narrower than hydrogen, all quantum defects used are taken from atomic database topbase. Illustration of stronger effects of ions and electrons on the alkaline profiles, than neutral-neutral collision mechanism. Keywords : Stars: fundamental parameters - Atomic processes - Line: profiles.

  18. Serum parameters predict the severity of ultrasonographicifndingsinnon-alcoholic fatty liver disease

    Mohsen Razavizade; Raika Jamali; Abbas Arj; Hamidreza Talari


    BACKGROUND: Controversy exists about the correlation between liver ultrasonography and serum parameters for evaluating the severity of liver involvement in non-alcoholic fatty liver disease (NAFLD). This study was designed to determine the association between liver ultrasonography staging in NAFLD and serum parameters correlated with disease severity in previous studies; and set optimal cut-off points for those serum parameters correlated with NAFLD staging at ultrasonography, in order to differentiate ultrasonographic groups (USGs). METHODS: This cross-sectional study evaluated outpatients with evidence of NAFLD in ultrasonography referred to a general hospital. Those with positive viral markers, abnormal serum ceruloplasmin or gamma-globulin concentrations were excluded. A radiologist performed the ultrasonography staging and stratiifed the patients into mild, moderate, and severe groups. Fasting serum alanine aminotransferase (ALT), aspartate aminotransferase, alkaline phosphatase, triglyceride (TG), high and low density lipoprotein (HDL, LDL), and cholesterol were checked. RESULTS:Two hundred and forty-ifve patients with a mean age (±standard deviation) of 41.63(±11.46) years were included. There were no signiifcant differences when mean laboratory concentrations were compared between moderate and severe USGs. Therefore, these groups were combined to create revised USGs ("mild"versus"moderate or severe"). There were associations between the revised USGs, and ALT, TG, HDL levels, and diabetes mellitus [odds ratios=2.81 (95%conifdence interval (CI):1.37-5.76), 2.48 (95%CI:1.29-4.78), 0.36 (95%CI:0.18-0.74), and 5.65 (95%CI:2.86-11.16) respectively;all P values CONCLUSIONS: Serum ALT, TG, and HDL concentrations seem to be associated with the staging by liver ultrasonography in NAFLD. They might be used to predict the staging of liver ultrasonography in these patients.

  19. Serum enzyme activities following long-distance running: comparison between Ethiopian and white athletes.

    Eliakim, A; Nemet, D; Shenkman, L


    Ethiopian runners are famous for their achievements in long-distance running. The recent immigration of Ethiopians to Israel provided an opportunity to compare some physiological variables between elite Ethiopian and white Israeli runners. Six Ethiopian and five white Israeli runners, aged 20 to 40 years, were studied before and after an 11 km race. Venous blood was sampled from each runner prior to the race, and 1 h, 48 h, and 5 days following the race. The activities of creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) were measured, and levels of serum electrolytes, urea, creatinine, phosphorus, albumin, cholesterol, and alkaline phosphatase were also determined. CK activity rose 2.6-fold with a peak 5 days after the race. LDH and AST levels rose as well (1.4-1.3-fold, respectively). Significant elevations also occurred in serum phosphorus, uric acid, and creatinine concentrations 1 h after the race. In contrast to previous studies in which higher enzyme activities were reported in blacks, we did not detect any difference in serum enzyme values between black and white runners.

  20. [False positive serum des-gamma-carboxy prothrombin after resection of hepatocellular carcinoma].

    Hiramatsu, Kumiko; Tanaka, Yasuhito; Takagi, Kazumi; Iida, Takayasu; Takasaka, Yoshimitsu; Mizokami, Masashi


    Measurements of serum concentrations of des-gamma-carboxy-prothrombin (PIVKA-II) are widely used for diagnosing hepatocellular carcinoma (HCC). Recently, when we evaluated the correlation of PIVKA-II between two commercially available PIVKA-II immunoassay kits (Lumipulse f vs. Picolumi) to introduce it in our hospital, false high values of PIVKA-II were observed in Lumipulse assay. Four(4%) of 100 serum samples showed false high values, and all of them were obtained from patients less than 2 month after curative resection of HCC. Examining additional 7 patients with HCC resection, serum samples from the 5 patients had the same trend. To elucidate the non-specific reaction by Lumipulse assay which utilized alkaline phosphatase (ALP) enzymatic reaction, inhibition assays by various absorbents such as inactive ALP and IgM antibodies were performed. Excess of inactive ALP reduced the high values of PIVKA-II. Note that anti-bleeding sheets (fibrinogen combined drug), which included bovine thrombin, were directly attached on liver of all patients with HCC resection in this study. As the sheets also contaminate ALP and probably produce IgM antibodies to ALP, the IgM may cross-react with anti-PIVKA-II antibodies directly. Taken together, it was suggested that produced antibodies against ALP derived from anti-bleeding sheets led false high values of PIVKA-II in the patients with HCC resection.

  1. Derivation and characterization of human embryonic germ cells: serum-free culture and differentiation potential.

    Hua, Jinlian; Yu, Haisheng; Liu, Sheng; Dou, Zhongying; Sun, Yadong; Jing, Xiaoqi; Yang, Chunrong; Lei, Anmin; Wang, Huayan; Gao, Zhimin


    This study examined the effects of a chemically defined culture medium supplement, knock-out serum replacement (KSR), on the growth and differentiation of human embryonic germ cells (hEgc) and found that the efficiency of the initial establishment of hEGC lines in KSR medium was significantly higher than in fetal calf serum (FCS) medium. The percentage of undifferentiated hEGC colonies growing in KSR medium was significantly higher than in FCS-based medium (P embryonic germ cell-like morphology. They showed normal and stable diploid karyotype and expressed alkaline phosphatase (AP), stage-specific embryonic antigens (SSEA) and other specific markers of pluripotent cells. In addition, hEGC could form simple and cystic embryoid bodies (EB) that consisted of various cell types including neural, epithelial and rhythmically beating cardiac cells, even sperm-like and oocyte-like cells. Tumour-like outgrowths were formed in nude mice and found to contain a variety of cell types, including uterine epithelium, adipocytes, squamous tissue and skin structures. In conclusion, an appropriate serum-free culture system has been developed for the establishment of hEGC lines. This may provide an in-vitro model to study differentiation and can be used as a potential source of therapy for infertility and regenerative medicine.

  2. The Antagonistic Effect of Mycotoxins Deoxynivalenol and Zearalenone on Metabolic Profiling in Serum and Liver of Mice

    Jian Ji


    Full Text Available Metabolic profiling in liver and serum of mice was studied for the combined toxic effects of deoxynivalenol (DON and zearalenone (ZEN, through gas chromatography mass spectrum. The spectrum of serum and liver sample of mice, treated with individual 2 mg/kg DON, 20 mg/kg ZEN, and the combined DON + ZEN with final concentration 2 mg/kg DON and 20 mg/kg ZEN for 21 days, were deconvoluted, aligned and identified with MS DIAL. The data matrix was processed with univariate analysis and multivariate analysis for selection of metabolites with variable importance for the projection (VIP > 1, t-test p value < 0.05. The metabolic pathway analysis was performed with MetaMapp and drawn by CytoScape. Results show that the combined DON and ZEN treatment has an obvious “antagonistic effect” in serum and liver tissue metabolic profiling of mice. The blood biochemical indexes, like alkaline phosphatase, alanine transaminase, and albumin (ALB/globulin (GLO, reveal a moderated trend in the combined DON + ZEN treatment group, which is consistent with histopathological examination. The metabolic pathway analysis demonstrated that the combined DON and ZEN treatment could down-regulate the valine, leucine and isoleucine biosynthesis, glycine, serine and threonine metabolism, and O-glycosyl compounds related glucose metabolism in liver tissue. The metabolic profiling in serum confirmed the finding that the combined DON and ZEN treatment has an “antagonistic effect” on liver metabolism of mice.

  3. The Antagonistic Effect of Mycotoxins Deoxynivalenol and Zearalenone on Metabolic Profiling in Serum and Liver of Mice

    Ji, Jian; Zhu, Pei; Cui, Fangchao; Pi, Fuwei; Zhang, Yinzhi; Li, Yun; Wang, Jiasheng; Sun, Xiulan


    Metabolic profiling in liver and serum of mice was studied for the combined toxic effects of deoxynivalenol (DON) and zearalenone (ZEN), through gas chromatography mass spectrum. The spectrum of serum and liver sample of mice, treated with individual 2 mg/kg DON, 20 mg/kg ZEN, and the combined DON + ZEN with final concentration 2 mg/kg DON and 20 mg/kg ZEN for 21 days, were deconvoluted, aligned and identified with MS DIAL. The data matrix was processed with univariate analysis and multivariate analysis for selection of metabolites with variable importance for the projection (VIP) > 1, t-test p value < 0.05. The metabolic pathway analysis was performed with MetaMapp and drawn by CytoScape. Results show that the combined DON and ZEN treatment has an obvious “antagonistic effect” in serum and liver tissue metabolic profiling of mice. The blood biochemical indexes, like alkaline phosphatase, alanine transaminase, and albumin (ALB)/globulin (GLO), reveal a moderated trend in the combined DON + ZEN treatment group, which is consistent with histopathological examination. The metabolic pathway analysis demonstrated that the combined DON and ZEN treatment could down-regulate the valine, leucine and isoleucine biosynthesis, glycine, serine and threonine metabolism, and O-glycosyl compounds related glucose metabolism in liver tissue. The metabolic profiling in serum confirmed the finding that the combined DON and ZEN treatment has an “antagonistic effect” on liver metabolism of mice. PMID:28075412

  4. Serum Predictors of Percent Lean Mass in Young Adults.

    Lustgarten, Michael S; Price, Lori L; Phillips, Edward M; Kirn, Dylan R; Mills, John; Fielding, Roger A


    Lustgarten, MS, Price, LL, Phillips, EM, Kirn, DR, Mills, J, and Fielding, RA. Serum predictors of percent lean mass in young adults. J Strength Cond Res 30(8): 2194-2201, 2016-Elevated lean (skeletal muscle) mass is associated with increased muscle strength and anaerobic exercise performance, whereas low levels of lean mass are associated with insulin resistance and sarcopenia. Therefore, studies aimed at obtaining an improved understanding of mechanisms related to the quantity of lean mass are of interest. Percent lean mass (total lean mass/body weight × 100) in 77 young subjects (18-35 years) was measured with dual-energy x-ray absorptiometry. Twenty analytes and 296 metabolites were evaluated with the use of the standard chemistry screen and mass spectrometry-based metabolomic profiling, respectively. Sex-adjusted multivariable linear regression was used to determine serum analytes and metabolites significantly (p ≤ 0.05 and q ≤ 0.30) associated with the percent lean mass. Two enzymes (alkaline phosphatase and serum glutamate oxaloacetate aminotransferase) and 29 metabolites were found to be significantly associated with the percent lean mass, including metabolites related to microbial metabolism, uremia, inflammation, oxidative stress, branched-chain amino acid metabolism, insulin sensitivity, glycerolipid metabolism, and xenobiotics. Use of sex-adjusted stepwise regression to obtain a final covariate predictor model identified the combination of 5 analytes and metabolites as overall predictors of the percent lean mass (model R = 82.5%). Collectively, these data suggest that a complex interplay of various metabolic processes underlies the maintenance of lean mass in young healthy adults.

  5. [Protein phosphatases: structure and function].

    Bulanova, E G; Budagian, V M


    The process of protein and enzyme systems phosphorylation is necessary for cell growth, differentiation and preparation for division and mitosis. The conformation changes of protein as a result of phosphorylation lead to increased enzyme activity and enhanced affinity to substrates. A large group of enzymes--protein kinases--is responsible for phosphorylation process in cell, which are divided into tyrosine- and serine-threonine-kinases depending on their ability to phosphorylate appropriate amino acid residues. In this review has been considered the functional importance and structure of protein phosphatases--enzymes, which are functional antagonists of protein kinases.

  6. Antioxidants, cadmium-induced toxicity, serum biochemical and the histological abnormalities of the kidney and testes of the male Wistar rats.

    Obianime, A W; Roberts, I I


    The effect of different doses of cadmium [CD] on some biochemical, hormonal and histopathological parameters of the liver, kidney and testes of the Wistar rate were investigated. Cadmium in the dose range 0-40 mg/kg while causing a time-and dose-dependent decrease of the basal serum levels of alkaline phosphatase [ALP] also caused a dose-dependent increase in the serum concentration of the acid and prostatic acid phosphatases. The value of the ALP changed from 148.7+/-1.0 IU/L in the control to 53.7+/-0.098 at 40 mg/kg of cadmium. While the ACP and ACPT changed from 32.6+/-0.72 and 7 Units in the control to 54 and 17 units respectively at 40 mg/kg of CD. Furthermore cadmium also caused positively correlated dose-and time-dependent destruction of the histology of the liver, kidney and testes. These were characterized by vascular congestion, vacuolation, destruction of the seminal epithelial layers, focal necrosis of nucleus, oedema of the seminal epithelia layers, focal necrosis of nucleus, oedema of the seminiferous tubules and reduction of spermatogenesis. CD also caused granular and eosinophilic cytoplasm, enlargement of sinusoids with kupffer cells, haemorrhage and apoptosis of cells. Finally pre-treatment with vitamin C [0.0015/kg], vitamin E [1.51/g] and selenium [0.25 mg] which on their own had little or no effects on the serum basal phosphatases, hormonal and histological stability caused a reversal of the cadmium-induced biochemical, hormonal and histological toxicities of the liver, kidney and testes. These results may be explained by the oxidational/antioxidational properties of these xenobiotics and their mechanisms of actions.

  7. 75 FR 80826 - Compliance Policy Guide Sec. 527.300 Dairy Products-Microbial Contaminants and Alkaline...


    ... HUMAN SERVICES Food and Drug Administration Compliance Policy Guide Sec. 527.300 Dairy Products... Compliance Policy Guide Sec. 527.300 Dairy Products-- Microbial Contaminants and Alkaline Phosphatase... other indicators of inadequate pasteurization or post-pasteurization contamination of dairy...

  8. Radiographic imaging of metabolic bone disorders in consonance with biochemical parameters

    Parveen Chandna


    Conclusions: In all the osteoporotic cases irrespective to age of patients, values of serum calcium, serum alkaline phosphatase, serum phosphorus, serum potassium, serum sodium, serum calcidiol were within normal limits.

  9. The salubrious effect of tamoxifen [correction of Tamaxifen] on serum marker enzymes, glycoproteins, and lysosomal enzymes level in breast cancer woman.

    Thangaraju, M; Rameshbabu, J; Vasavi, H; Ilanchezhian, S; Vinitha, R; Sachdanandam, P


    Tumour markers correlate strongly with prognosis based on tumour burden and surgical resectability. If chemotherapy is extremely effective in certain stage of the disease, the sensitive marker may be of great use in monitoring disease response and drug treatment. Hence, this study was launched to evaluate the changes in tumour marker enzymes like lactate dehydrogenase (LDH), glutamate oxaloacetate transaminase (SGOT), glutamate pyruvate transaminase (SGPT), alkaline phosphatase, and acid phosphatase in before and after 3 and 6 months tamoxifen treated breast cancer patients. In addition, the changes in serum glycoproteins viz., hexose, hexosamine, and sialic acid and lysosomal enzymes such as N-acetyl-beta-D-glucosaminidase, beta-D-galactosidase, and beta-D-glucuronidase were analysed in these patients. These values were compared with their age matched healthy control subjects. At 6 months evaluation, the tamoxifen treated postmenopausal breast cancer women showed a statistically significant decreased (p sialic acid and N-acetyl-beta-D-glucosaminidase, beta-D-galactosidase, and beta-D-glucuronidase were decreased significantly (p breast cancer as well as the bone demineralisation in postmenopausal breast cancer women. Thus, tamoxifen may also have its antitumour activity through its beneficial effects on tumour marker enzymes and serum proteins in breast cancer women.

  10. 骨碱性磷酸酶 N-端骨钙素99Tcm-亚甲基二膦酸盐骨显像与早期骨代谢改变%The change of bone alkaline phosphatase,N-osteocalcin and 99Tcm-MDP bone imaging on bone metabolism in early stage

    罗裕旋; 吴传安; 杨伟康; 何小媚; 魏建伟; 梁瑞金; 余树林


    目的:探讨骨碱性磷酸酶(BALP)、N-端骨钙素(BGP)、99 Tc m-亚甲基二膦酸盐(MDP)骨显像与早期骨代谢改变的关系。方法通过注射地塞米松磷酸钠针剂(DX)得到骨质疏松兔,注射时间为6周。将兔分为3组,正常对照组、骨质疏松对照组(DX 注射量为2 mg/kg,1周2次)、骨量减少组(DX 注射量为1 mg/kg,1周1次)。分别检测3组的核素99 Tc m-MDP 骨显像、病理切片骨组织学、骨密度以及血清 BALP 与 BGP。对第3腰椎、膝关节、股骨头、股骨中段以及第4尾椎进行感兴区(ROI)半定量检测作比较。结果骨质疏松对照组与骨量减少组相比,在核素骨显像中只有股骨中段对99 Tc m-MDP 摄取的 ROI 比值上升(P 0.05);但骨密度以及血清 BALP 差异具有统计学意义(P 0.05).But the bone density and serum BALP results showed significant difference (P <0.05).Conclusion 9 9 Tc m-MDP bone imaging showed,the ROI ratio of femoral head,third lumbar,knee joint and fourth tail verte-brae ROI are increased.The ROI ratio in the middle part of the femur is not increased,it might be the time of bone mass reduction,the BGP support conclusion is more meaningful,and can provide reference for early preven-tion and treatment.

  11. 饲料锌含量对方格星虫稚虫生长性能、体成分、体腔液中锌含量及碱性磷酸酶活性的影响%Effects of Dietary Zinc Content on Growth Performance, Body Composition, Coelomic Fluid Zinc Content and Alkaline Phosphatase Activity of Juvenile Peanut Worm, Sipunculus nudus Linnaeus

    许明珠; 张琴; 童潼; 董兰芳; 杨家林; 蒋艳; 黄国强


    This trial was conducted to investigate the effects of dietary zinc content on growth performance, body composition, coelomic fluid zinc content and alkaline phosphatase ( AKP ) activity of juvenile peanut worm, Sipunculus nudus Linnaeus ( S. nudus ) . Juvenile peanut worm with the average body weight of (14.54±0.10) mg were fed six isonitrogenous and isoenergetic microparticle diets with different supplemental levels of zinc ( zinc methionine as zinc source) for 8 weeks, and the measured zinc contents in diets were 9.3, 31.7, 49.9, 90.1, 168.6 and 326.5 mg/kg, respectively. Each diet had three replicates and each replicate had 400 juvenile peanut worm. The results showed that the dietary zinc content had significant effects on weight gain rate ( WGR) , specific growth rate ( SGR) and survival ratio ( SR) of S. nudus ( P0.05). Dietary zinc content significantly affected coelomic fluid zinc content and AKP activity of S. nudus ( P<0.05) . With the increase of dietary zinc content, the coelomic fluid zinc content of S. nudus was sustained rise, when the dietary zinc content was 326.5 mg/kg, it produced the maximum value. Moreover, the coelomic fluid AKP activity was firstly increased and then tended to be stable, when the dietary zinc content was 49.9 mg/kg, it produced the maximum value. With WGR as the evaluating index, the suitable dietary zinc content of juvenile peanut worm is 41.93 mg/kg by regression analysis.%以蛋氨酸锌为锌源,在等氮等能的基础饲料中添加不同水平的锌,配制实际锌含量分别为9.3、31.7、49.9、90.1、168.6和326.5 mg/kg 的6种试验饲料,饲喂平均体重为(14.54±0.10) mg的方格星虫稚虫8周,用以研究饲料锌含量对方格星虫稚虫生长、体成分、体腔液中锌含量及碱性磷酸酶活性的影响。每种试验饲料设3个重复,每个重复饲喂400尾方格星虫稚虫。结果表明:饲料锌含量对方格星虫的成活率、增重率和特定

  12. Biomineralization of Uranium by PhoY Phosphatase Activity Aids Cell Survival in Caulobacter crescentus

    Yung, M C [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Jiao, Y [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)


    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  13. Biomineralization of uranium by PhoY phosphatase activity aids cell survival in Caulobacter crescentus.

    Yung, Mimi C; Jiao, Yongqin


    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  14. An alkaline element

    Arita, T.; Murakami, K.; Okha, K.


    A cathode with a dual layer active mass is installed in the disk shaped alkaline silver and zinc element. The first layer, which is turned towards the anode, contains 85 parts Ag2O, 5 parts electrolytic MnO2 and 10 parts graphite. The second layer, which contacts the bottom of the element, contains 35 parts Ag2O, 60 parts electrolytic MnO2 and 5 parts graphite. The electrical capacity of the first and second layers is 60 and 40, respectively. The first layer may be discharged with a high current density and the second layer with less current density. The element has high characteristics with comparatively low cost.

  15. A Study to Assess the Validity of Estimation of Serum Ostase Level in Hyperthyroid and Hypothyroid Cases

    Boruah, Polina; Hajong, Ranendra; Nath, Chandan Kumar; Barman, Bhupen; Chutia, Happy; Sarma, Kalyan


    Introduction One of the more specific assessments of the metabolic status of bone in normal and in disease conditions is the measurement of bone specific alkaline phosphatase or ostase. The measurement of serum ostase has several advantages over the measurements of other bone parameter. Because of its relatively long half-life, in-vivo (1 to 3days), it is relatively unaffected by diurnal variation. Aim To find the correlation of serum ostase level in hyper and hypothyroid cases and also to study the validity of routine estimation of serum ostase in hyper and hypothyroid cases so as to monitor the base level bone health on presentation. Materials and Methods Serum ostase level was studied in 74 patients with disorder of thyroid function. Serum ostase level, Thyroid Stimulating Hormone (TSH), FT3, FT4 levels were estimated by chemiluminescent technique. The instrument used was Beckman- coulter Access 2. A total of 39 patients were hypothyroid, 31 were hyperthyroid and 4 patients had subclinical hyperthyroidism. Results The serum ostase level was found to be elevated above 40 μg/L in 26 of the cases and above 16 μg/L but below 40μg/L in 5 cases of hyperthyroidism along with decrease in Bone Mineral Density (BMD). Serum ostase level was found to be directly proportional to the serum FT3 level (Normal range of serum ostase is 8-16 μg/L). Conclusion From this study, an inference can be drawn that a routine estimation of serum ostase level in hyperthyroid cases will help in proper monitoring of decrease bone turnover as indicated by increase serum ostase level. Besides, the estimation of serum ostase level in hyperthyroid cases it is found to be valid in this study, which can turn to be an important guiding parameter to the treating physician to formulate necessary protocols and guidelines for prophylaxis, treatment and to monitor the response to therapy in cases of reduced bone turnover related to hyperthyroid state. PMID:27790421

  16. Diagnostic value of prostatic acid phosphatase as determined by radioimmunoassay

    Wirth, M.P.; Osterhage, H.R.; Ackkermann, R.


    Serum concentrations of prostatic acid phosphatase (PAP) were determined with 4 different radioimmunoassays and with the standard enzymatic method (p-nitrophenylphosphate) in 35 patients with prostatic carcinoma. Staging of localized tumors was based on histopathological evaluation after radial prostatectomy and pelvic lymphnode dissection (pTsub(1-3), pN/sub 0/). In tumor lesions Tsub(1-2) N/sub 0/ M/sub 0/ elevated PAP-serum concentrations were found by RIA-determination in only one patient. Increased PAP serum levels were observed in 43-78% of carcinomas stage T/sub 3/ N/sub 0/ M/sub 0/ and in 54-83% in stage Tsub(2-4) Nsub(x) M/sub 1/ tumors, depending on the test kit used for the PAP determination. Concentrations for PAP obtained with the 4 different RIA-kits used, varied significantly and thus are not comparable. No false positive results were observed in sera of 9 patients with benign prostatic hyperplasia. Elevated PAP serum levels were found in a significantly higher frequency when determined by radioimmunoassay than by the enzymatic method. The results clearly indicate, that PAP is of no value for early recognition of carcinoma of the prostate even when measured by radioimmunoassay. However, the RIA-method seems to be of clinical importance in estimating the course of advanced local and metastasizing carcinoma of the prostate.

  17. Effects of Dietary Zinc Oxide and a Blend of Organic Acids on Broiler Live Performance, Carcass Traits, and Serum Parameters

    BG Sarvari


    Full Text Available ABSTRACT This experiment was carried out to evaluate the effect of different dietary supplementation levels of zinc oxide and of an organic acid blend on broiler performance, carcass traits, and serum parameters. A total of 2400 one-day-old male Ross 308 broiler chicks, with average initial body weight 44.21±0.19g, was distributed according to a completely randomized design in a 2 x 3 factorial arrangement. Six treatments, consisting of diets containing two zinc oxide levels (0 and 0.01% of the diet and three organic acid blend levels (0, 0.15, and 0.30% were applied, with eight replicates of 50 birds each. The experimental diets were supplied ad libitum for 42 days. There were significant performance differences among birds fed the different zinc oxide and organic acid blend levels until 42 d of age (p<0.01. The result of this experiment showed that the organic acid blend did not affect feed intake, but zinc oxide increased feed intake. Carcass traits were not influenced by the experimental supplements. Zinc oxide supplementation increased serum alkaline phosphatase level (p<0.01. The organic acid blend reduced serum cholesterol and triglyceride levels (p<0.05. No interactions were found between zinc oxide and the organic acid blend for none of the evaluated parameters. We concluded that zinc oxide and the evaluated organic acid blend improve broiler performance.

  18. Comparative study of haematological, serum electrolyte and nonelectrolyte parameters of male and female Persian sturgeon (Acipenser persicus) brood stocks

    ADEL Milad; PALANISAMY Satheesh Kumar; SHAFIEI Shafigh; FAZLI Hasan; ZORRIEHZAHRA Mohammad Jalil


    Sturgeons are the most important principal market species in the Caspian Sea. In the present study, measurement of blood parameters was carried out with the aim of evaluating the normal value of hematological and serum biochemical parameters of brood stocksAcipenser persicus during spawning season. Blood samples were collected from 36 brood stocks ofA. persicus (18 males and 18 females) during the spawning season. Higher value of red blood cells, white blood cells, haemoglobin, haematocrit was observed in male breeders with significant differences between female breeders (p>0.05). Haemoglobin and haematocrit value in this study was within the range of 8.70 to 9.2 g/dL and 29.73% to 30.26%, respectively. The statistically significant differences between mean corpuscular volumes, mean corpuscular haemoglobin, lymphocyte and neutrophil percentages, aspartate aminotransferase, alkaline phosphatase and creatine kinase were observed in the male and female brood stocks. It is concluded from this study, some of the hematological and serum biochemical parameters of male and female A. persicus breeders were significantly varied from each other. In comparative investigation with earlier reports, the values of red blood cells, haemoglobin and haematocrit are highly varied due to age variations of fish. However, further studies are necessary to understand the impact of sexual, age, size, length, and season and habitat variation. Serum biochemical parameters can be used for confirming the maturity ofA. persicus and monitoring any changes in the water quality parameters and soil.

  19. An elevated serum miR-141 level in patients with bone-metastatic prostate cancer is correlated with more bone lesions

    Hai-Liang Zhang; Xiao-Jian Qin; Da-Long Cao; Yao Zhu; Xu-Dong Yao; Shi-Lin Zhang; Bo Dai


    The skeleton is the most common metastatic organ in patients with prostate cancer (PCa).Non-invasive biomarkers that can facilitate the detection and monitoring of bone metastases are highly desirable.We designed this study to assess the expression patterns of serum miR-141 in patients with bone-metastatic PCa.Serum samples were collected to measure the miR-141 level in 56 patients,including six with benign prostatic hyperplasia (BPH),20 with localized PCa and 30 with bone-metastatic PCa (10 with hormone-naive PCa,10 with hormone-sensitive PCa and 10 with hormone-refractory PCa).A bone scan was performed for each patient with PCa to assess the number of bone lesions.The quantification of serum miR-141 levels was assayed by specific TaqMan qRT-PCR.The results showed that serum miR-141 levels were elevated in patients with bone metastasis (P<0.001).There was no statistically significant difference in the serum miR-141 levels between patients with BPH and patients with localized PCa.Using Kendall's bivariate correlation test,both the Gleason score and the number of bone-metastatic lesions were found to correlate with serum miR-141 levels (P=0.012 and P<0.001,respectively).The serum miR-141 level was found to be positively correlated with alkaline phosphatase (ALP) level in patients with skeletal metastasis,using Pearson's bivariate correlation test.No relationship was found between the serum miR-141 level and the serum prostate-specific antigen (PSA) level.We concluded that serum miR-141 levels are elevated in patients with bone-metastatic PCa and that patients with higher levels of serum miR-141 developed more bone lesions.Furthermore,serum miR-141 levels are correlated with serum ALP levels but not serum PSA levels.

  20. Serum biochemistry of captive and free-ranging gray wolves (Canis lupus)

    Constable, Peter; Hinchcliff, Ken; Demma, Nick; Callahan, Margaret; Dale, B.W.; Fox, Kevin; Adams, Layne G.; Wack, Ray; Kramer, Lynn


    Normal serum biochemistry values are frequently obtained from studies of captive sedentary (zoo) or free-ranging (wild) animals. It is frequently assumed that values from these two populations are directly referable to each other. We tested this assumption using 20 captive gray wolves (Canis lupus) in Minnesota, USA, and 11 free-ranging gray wolves in Alaska, USA. Free-ranging wolves had significantly (P<0.05) lower sodium, chloride, and creatine concentrations and significantly higher potassium and blood urea nitrogen (BUN) concentrations; BUN to creatine ratios; and alanine aminotransferase, aspartate aminotransferase, and creatine kinase activities relative to captive wolves. Corticosteroid-induced alkaline phosphatase activity (a marker of stress in domestic dogs) was detected in 3 of 11 free-ranging wolves and in 0 of 20 captive wolves (P = 0.037). This study provides clear evidence that serum biochemical differences can exist between captive and free-ranging populations of one species. Accordingly, evaluation of the health status of an animal should incorporate an understanding of the potential confounding effect that nutrition, activity level, and environmental stress could have on the factor(s) being measured.

  1. Changes in selected hematology and serum biochemistry in Turkish Angora cats (Felis catus during growth period

    Ozkan Simsek


    Full Text Available The purpose of the present study was to determine the changes in selected hematology and serum biochemistry of Angora cats (Felis catus during growth period. A total of 32 Angora cats (16 adults and 16 kittens were used in this study. Blood samples were collected from the animals, and were analyzed for white blood cells, red blood cells, hemoglobin, packed cell volume, mean corpuscular volume (MCV, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, granulocytes, monocytes and lymphocytes numbers. In the serum, alanine aminotransferase, aspartate aminotransferase (AST, alkaline phosphatase (ALP, gamma glutamyl transferase (GGT, lactate dehydrogenase (LDH, creatinine kinase (CK, total cholesterol, glucose, triglyceride, urea, creatinine, total protein, albumin, Ca, Mg, Pi levels were determined. Monocyte level was found higher, and ALP, LDH, CK activities and Pi levels were lower in adult cats as compared to the kittens. MCV was lower and GGT and AST activities, and glucose level were higher in kittens of 1.5-3 months old than in kittens of >3 months. Concentrations of total cholesterol and Mg were higher in kitten (1.5-3 months old than in adult cats. In conclusion, age related effects on hematological and biochemical blood parameters have been determined for the first time in Angora cats.

  2. Relationships between serum Omentin-1 levels and bone mineral density in older men with osteoporosis

    Li Yang; Xin-Lan Zhao; Bin Liao; Ai-Ping Qin


    Objective: To investigate the correlation between serum Omentin-1 levels and the presence of osteoporosis in older men. Methods: Serum Omentin-1, bone turnover biochemical markers, and bone mineral density (BMD) were determined in 45 older men with osteoporosis or 45 older men without osteoporosis (65e70 years old). Results: Omentin-1 levels were increased in older men with osteoporosis, and the differences remained significant after con-trolling for fat mass. Omentin-1 was negatively correlated with BMD. In a multiple linear stepwise regression analysis, Omentin-1, lean mass, but not fat mass, were independent predictors of BMD for the combined group. Significant negative correlations between Omentin-1 and bone-specific alkaline phosphatase (BAP) and bone cross-linked N-telopeptides of typeⅠcollagen (NTX) were found. Omentin-1 was also independently associated with BMD and bone turnover markers in older men with osteoporosis and control groups that were considered separately. Conclusions: Omentin-1 is an independent predictor of BMD in older men with osteoporosis, and it is negatively correlated with bone turnover biochemical markers. It is suggested that Omentin-1 may exert a negative effect on bone mass through the regulation of the osteoblast differentiation in the older men with osteoporosis.

  3. Bioprecipitation of uranium from alkaline waste solutions using recombinant Deinococcus radiodurans

    Kulkarni, Sayali; Ballal, Anand; Apte, Shree Kumar, E-mail:


    Highlights: • Deinococcus radiodurans was genetically engineered to overexpress alkaline phosphatase (PhoK). • Deino-PhoK bioprecipitated U efficiently over a wide range of input U concentration. • A maximal loading of 10.7 g U/g of biomass at 10 mM input U was observed. • Radioresistance and U precipitation by Deino-PhoK remained unaffected by γ radiation. • Immobilization of Deino-PhoK facilitated easy separation of precipitated U. -- Abstract: Bioremediation of uranium (U) from alkaline waste solutions remains inadequately explored. We engineered the phoK gene (encoding a novel alkaline phosphatase, PhoK) from Sphingomonas sp. for overexpression in the radioresistant bacterium Deinococcus radiodurans. The recombinant strain thus obtained (Deino-PhoK) exhibited remarkably high alkaline phosphatase activity as evidenced by zymographic and enzyme activity assays. Deino-PhoK cells could efficiently precipitate uranium over a wide range of input U concentrations. At low uranyl concentrations (1 mM), the strain precipitated >90% of uranium within 2 h while a high loading capacity of around 10.7 g U/g of dry weight of cells was achieved at 10 mM U concentration. Uranium bioprecipitation by Deino-PhoK cells was not affected in the presence of Cs and Sr, commonly present in intermediate and low level liquid radioactive waste, or after exposure to very high doses of ionizing radiation. Transmission electron micrographs revealed the extracellular nature of bioprecipitated U, while X-ray diffraction and fluorescence analysis identified the precipitated uranyl phosphate species as chernikovite. When immobilized into calcium alginate beads, Deino-PhoK cells efficiently removed uranium, which remained trapped in beads, thus accomplishing physical separation of precipitated uranyl phosphate from solutions. The data demonstrate superior ability of Deino-PhoK, over earlier reported strains, in removal of uranium from alkaline solutions and its potential use in

  4. Total serum bile acid as a potential marker for the diagnosis of cholangiocarcinoma without jaundice.

    Sombattheera, Sutthikan; Proungvitaya, Tanakorn; Limpaiboon, Temduang; Wongkham, Sopit; Wongkham, Chaisiri; Luvira, Vor; Proungvitaya, Siriporn


    Diagnosis of cholangiocarcinoma (CCA) is difficult when patients do not show jaundice. The aim of this study was to examine the feasibility of using the total serum bile acid (TSBA) level as an aid for the diagnosis of CCA in patients without jaundice. For this purpose, TSBA of the following groups were measured using a Beckman Synchron CX4 clinical chemistry analyzer: 60 cases of CCA with total serum bilirubin ≤2 mg/dL (low total bilirubin group, LTB); 32 cases of CCA with total serum bilirubin >2 mg/dL (high total bilirubin group, HTB); and 115 healthy controls. Liver function parameters such as serum cholesterol, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were also examined. The results showed that the TSBA of both LTB and HTB groups of the CCA patients were significantly higher than that of the healthy controls. Also, significant correlation was observed between TSBA and total bilirubin levels in the HTB group of CCA patients. However, no such correlation was seen in the LTB group. The cut-off value of TSBA was determined for the LTB group of CCA patients using the receiver operating characteristic curve analysis, and it was 6.05 μmol/L with the sensitivity and specificity of 46.7% and 84.4%, respectively. In addition, the ALP level was correlated well with the TSBA level and ALP in HTB group was significantly higher than that of LTB group. Moreover, the combination of high TSBA and high ALP levels gave higher specificity up to 97.4%. TSBA might be useful for the diagnosis of CCA patients without jaundice.

  5. Xeno-free culture condition for human bone marrow and umbilical cord matrix-derived mesenchymal stem/stromal cells using human umbilical cord blood serum

    Esmaeli, Azadeh; Moshrefi, Mojgan; Shamsara, Ali; Eftekhar-vaghefi, Seyed Hasan; Nematollahi-mahani, Seyed Noureddin


    Background: Fetal bovine serum (FBS) is widely used in cell culture laboratories, risk of zoonotic infections and allergic side effects create obstacles for its use in clinical trials. Therefore, an alternative supplement with proper inherent growth-promoting activities is demanded. Objective: To find FBS substitute, we tested human umbilical cord blood serum (hUCS) for proliferation of human umbilical cord matrix derived mesenchymal stem cells (hUC-MSCs) and human bone marrow-derived mesenchymal cells (hBM-MSCs). Materials and Methods: Umbilical cord blood of healthy neonates, delivered by Caesarian section, was collected and the serum was separated. hUC-MSCs and hBM-MSCs were isolated and characterized by assessment of cell surface antigens by flow cytometry, alkaline phosphatase activity and osteogenic/adipogenic differentiation potential. The cells were then cultured in Iscove's Modified Dulbecco's Medium (IMDM) by conventional methods in three preparations: 1- with hUCS, 2- with FBS, and 3- without serum supplements. Cell proliferation was measured using WST-1 assay, and cell viability was assessed by trypan blue staining. Results: The cells cultured in hUCS and FBS exhibited similar morphology and mesenchymal stem cells properties. WST-1 proliferation assay data showed no significant difference between the proliferation rate of either cells following hUCS and FBS supplementation. Trypan blue exclusion dye test also revealed no significant difference for viability between hUCS and FBS groups. A significant difference was detected between the proliferation rate of stem cells cultured in serum-supplemented medium compared with serum-free medium. Conclusion: Our results indicate that human umbilical cord serum can effectively support proliferation of hBM-MSCS and hUC-MSCs in vitro and can be used as an appropriate substitute for FBS, especially in clinical studies. PMID:27738658

  6. 晚期妊娠妇女血清α-L-岩藻糖苷酶和碱性磷酸酶检测及临床意义%Detaction serum alpha-L-fucosidase, alkaline phosphatase and its clinical significance for women in late pregnancy

    王婧婧; 王文燕; 常永超


    目的 测定晚期妊娠妇女血清中α-L-岩藻糖苷酶和碱性磷酸酶的含量,以探讨其临床意义.方法 实验组169例妊娠期妇女,对照组137例健康育龄妇女,分别检测血清AFU、ALP含量.结果 实验组血清AFU、ALP含量较对照组高,差异有统计学意义(P<0.01),且随孕周数增加AFU、ALP也有所增加.结论 血清AFU、ALP含量测定可以作为晚期妊娠妇女监测胎儿、胎盘的指标.

  7. Phosphate solubilizing bacteria and alkaline phosphatase activity in coastal waters off Trivandrum

    Mamatha, S.S.; Gobika, A.; Janani, P.

    responds to increase in temperature measured in these waters. Our studies suggest that the activity off coastal waters is stimulated by fall in levels of ambient phosphate concentrations. Detailed studies would help us to establish the threshold...

  8. Construction of single chain Fv antibody against transferrin receptor and its protein fusion with alkaline phosphatase

    Dao-Feng Yang; Hui-Fen Zhu; Zhi-Hua Wang; Guan-Xin Shen; De-Ying Tian


    AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by SfiⅠ and NotⅠ, it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E. coli TG1. The positive colonies were screened by colony PCR and their expressions were induced by IPTG. ScFv gene was gained by digesting ScFv expression vector pUC19/119 with Sfi I and NotⅠ restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E. coli TG1. The positive colonies were selected by bacterial colony PCR. The expression of fusion protein (scFv-AP) was induced by IPTG. Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku. Immunofluorescent assay (IFA) demonstrated its reactivity with TfR. The molecular weight of scFv-AP was 75 ku. Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity.CONCLUSION: We have successfully prepared the antihuman TfR scFv and constructed the fusion protein of scFv and AP. It is promising for immunological experiments.

  9. Amino acid sequence of the cold-active alkaline phosphatase from Atlantic cod (Gadus morhua)

    Asgeirsson, Bjarni; Nielsen, Berit Noesgaard; Højrup, Peter


    -linked glycosylation sites were found. The glycan structure was determined as complex biantennary in type with fucose and sialic acid attached, although a trace of complex tri-antennary structure was also observed. A three-dimensional model was obtained by homology modelling using the human placental AP scaffold. Cod...

  10. In situ hybridization of cytokine mRNA using alkaline phosphatase-labelled oligodeoxynucleotide probes

    Clausen, Bettina Hjelm; Fenger, Christina; Finsen, B.


    In situ hybridization is a powerful tool for visualizing cellular gene expression in morphologically preserved brain tissue giving precise information on the regional expression of specific mRNA sequences in cells of diverse phenotype. Here, we describe a sensitive, simple, and robust method using...

  11. Alkaline phosphatase predicts relapse in chronic hepatitis C patients with end-of-treatment response

    Gerd; Bodlaj; Rainer; Hubmann; Karim; Saleh; Tatjana; Stojakovic; Georg; Biesenbach; Jrg; Berg


    AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genoty...

  12. The significance of amperometric detection of alkaline phosphatase in colorectal cancer diagnostics

    Belkin, Anton; Freynd, Genrietta; Katsnelson, Mikhail


    Colorectal cancer (CRC) is one of the most common cancers in the world; it takes the second place in oncological morbidity. The ALP activity of intestinal epithelial differentiation marker (ALP) was investigated in surgical material and colon biopsies of 47 patients with the electrochemical method using biosensors (biochips). The average current obtained in the studies of colorectal cancer tissues was lower than in the studies of intact colon mucosa. Histology of tumors matched the differentiation types/stages of adenocarcinomas. The study of ALP activity in the surgical material and biopsies of colon tumors can become one of the most useful methods for evaluating the functional atypia in tumor tissue.

  13. Quantitation of Alkaline Phosphatase Isoenzymes Using Agarose Containing Wheat Germ Lectin


    Trizma (50 mmol)/barbital (14 mmol)/sodium barbital (50 mmol) buffer at pH 8.9 is available prepackaged from Sigma (710-1). Dissolve one 10 g vial in...materials. The separation of AP isoenzymes requires that the buffer pH be maintained at 8.9 +0.1. Electrophoresis 53 54 conducted using trizma ...21 Electrophoresis Buffer ........................... 21 AMP-Metal Ion Buffer ............................. 21 Agarose

  14. The Effects of Short-Term Intensive Exercise on Levels of Liver Enzymes and Serum Lipids in Kick Boxing Athletes

    Ömer Kaynar


    Full Text Available Objective: In this study, it was aimed to evaluate the ef­fects of short-term intensive exercise on liver enzymes and serum lipid levels with kick boxing athletes. Methods: 23 voluntary athletes who were between the ages of 15-46 and who engaged in kick–boxing have tak­en place this study. Athletes were made to do 45 minutes of warming-up, breathing, and stretching and 50 minutes of technical and tactical practices and then they were made to do a training match, which is equal to a 2 min­utes 3 circuits (1 minute rest kick-box match. In venous blood samples which were taken from athletes before and after training, aspartate aminotransferase (AST, alanine aminotransferase (ALT, alkaline phosphatase (ALP and gamma glutamine transpeptidase (GGT, enzyme activity and total cholesterol, high-density lipoprotein-cholesterol (HDL-C, low density lipoprotein-cholesterol (LDL-C and triglycerides serum levels were analyzed via spectropho­tometric method in Beckman Coulter AU 5800 auto ana­lyzer. Body composition measurements of athletes were made with Tanita TBF 300 brand device, which works with bio-impedance analysis (BIA system. Results: As a result of our study, statistically increases in serum ALT, AST, ALP and GGT enzyme activities and in serum total cholesterol, HDL-C and LDL-C levels were detected following short-term intensive exercise, but no significant difference was observed in TG levels after in­tensive exercise. Conclusion: The blows to the abdomen during kickbox­ing sports competitions result in increased liver enzymes and increased serum lipids may occur to meet energy de­mand of the body during exercise.

  15. The influence of starvation and water deprivation upon certain serum enzyme activity in chicks in the first week after hatching

    Markovska-Gosic Bozidarka


    Full Text Available In the past decade the determination of enzyme activity in blood serum, organs and tissues gained an innocuous meaning in the veterinary medicine, especially in the early stages of diagnosis of different metabolism abnormalities. However, the experience gained from the human medicine, without a detail analyses could not be applied on domestic animals and poultry, because of the fact that the enzyme activity in the cells of different animal tissues and organs differs from those of the humans. This explains the different diagnostic meaning of the enzyme activity in animals and humans. Up to date, an extensive research of the enzyme status is done, in order to gain information for their importance in the biochemical processes in the organism, disturbances in the cell membrane transport, disease diagnosis and nutritional disorders.The influence of different stress factors and conditions upon enzyme activity both in serum and tissues is still one of the major topics in many researches. In the first week after hatch important physiological changes occur in chick organism. In the same time, in the first 24-48 hours, they are exposed to the influence of different stressors such as transportation to the hatchery along with feed and water deprivation, what gives the assumption that changes do occur in the serum enzyme activity in the first days after hatch. As literature data on the influence of food and water deprivation upon serum enzyme activity are poor and often very contradictory, this paper will try to contribute to the extensive research on the topic and clear some misunderstandings. For that purpose serum activities of the following enzymes were analysed: alanine aminotransferase, aspartat aminotarnsferase, gamma glutamile transferase, actate dehydrogenase and alkaline phosphatase.

  16. Phosphatase activity in sandy soil influenced by mycorrhizal and non-mycorrhizal cover crops

    Alceu Kunze


    Full Text Available Cover crops may difffer in the way they affect rhizosphere microbiota nutrient dynamics. The purpose of this study was to evaluate the effect of mycorrhizal and non-mycorrhizal cover crops on soil phosphatase activity and its persistence in subsequent crops. A three-year experiment was carried out with a Typic Quartzipsamment. Treatments were winter species, either mycorrhizal black oat (Avena strigosa Schreb or the non-mycorrhizal species oilseed radish (Raphanus sativus L. var. oleiferus Metzg and corn spurry (Spergula arvensis L.. The control treatment consisted of resident vegetation (fallow in the winter season. In the summer, a mixture of pearl millet (Pennisetum americanum L. with sunnhemp (Crotalaria juncea L. or with soybean (Glycine max L. was sown in all plots. Soil cores (0-10 cm and root samples were collected in six growing seasons (winter and summer of each year. Microbial biomass P was determined by the fumigation-extraction method and phosphatase activity using p-nitrophenyl-phosphate as enzyme substrate. During the flowering stage of the winter cover crops, acid phosphatase activity was 30-35 % higher in soils with the non-mycorrhizal species oilseed radish, than in the control plots, regardless of the amount of P immobilized in microbial biomass. The values of enzyme activity were intermediate in the plots with corn spurry and black oat. Alkaline phosphatase activity was 10-fold lower and less sensitive to the treatments, despite the significant relationship between the two phosphatase activities. The effect of plant species on the soil enzyme profile continued in the subsequent periods, during the growth of mycorrhizal summer crops, after completion of the life cycle of the cover crops.

  17. Purification, enzymatic properties, and active site environment of a novel manganese(III)-containing acid phosphatase.

    Sugiura, Y; Kawabe, H; Tanaka, H; Fujimoto, S; Ohara, A


    A new manganese-containing acid phosphatase has been isolated and crystallized from sweet potato tubers. The pure enzyme contains one atom of manganese per Mr = 110,000 polypeptide and shows phosphatase activity toward various phosphate substrates. The pH optimum of the enzyme was 5.8 and the enzyme activity was inhibited by Cu2+, Zn2+, Hg2+, AsO43-, and MoO42-. This stable metalloenzyme is red-violet in color with an intense absorption band at 515 nm (epsilon - 2460). Our electronic, circular dichroism, and electron spin resonance findings strongly indicate that the Mn-valence state of the native enzyme is trivalent. When the Mn-enzyme is excited by the 5145 A line of Ar+ laser, prominent Raman lines at 1230, 1298, 1508, and 1620 cm-1 were detected. This Raman spectrum can probably be interpreted in terms of internal vibration of a coordinated tyrosine phenolate anion. The tryptophan-modified enzyme showed a positive Raman band at 370 cm-1, which is preferentially assigned to a Mn(III)-S streching mode. The modification of the Mn-enzyme by N-bromosuccinimide led to a large decrease in the fluorescence intensity of 335 nm which was dominated by its tryptophan residues within a considerable hydrophobic environment. The acid phosphatase activity was significantly decreased by the tryptophan modification. With respect to the active site donor sets, the Mn(III)-containing acid phosphatase is distinctly different from the Zn(II)-containing alkaline phosphatase. Of interest is also the appreciable similarity of some enzymatic and spectroscopic properties between the present enzyme and uteroferrin.

  18. Serum and seminal markers in the diagnosis of disorders of the genital tract of the dog: a mini-review.

    Gobello, Cristina; Castex, Gervasio; Corrada, Yanina


    Serum and seminal biologic substances that are produced either by normal or abnormal tissues of the organism and that can be used to diagnose pathological conditions are usually referred as markers. The aim of this article is to briefly review the most relevant clinical features of the main genital markers in the male dog: alkaline phosphatase (AP), carnitine and canine prostate-specific arginine esterase (CPSE). Carnitine and AP are markers for the presence of epididymal fluid in the ejaculate and their measurement in azoospermic dogs has been used as an indicator of tubular patency of the ductal network. Although AP is not present in high concentrations in the testis, this does not preclude the possibility that testicular cells might secrete some AP. If this were true, AP could also reflect, at least in some degree, germ cell function in this species. Prostate-specific arginine esterase, the major secretory product of the canine prostate, is a known marker of gland secretion in the dog. Tumor markers frequently used in human medicine, such as prostatic acid phosphatase and prostate-specific antigen, are is still controversial in the diagnosis of prostatic carcinoma of the dog. Although further research is necessary to define the exact role of CPSE, it seems to be a promising diagnostic tool in nonneoplasic canine prostatic disorders. Future studies should also address the quantitative relationship among serum and prostatic androgen levels, prostatic androgen-dependent problems and how these are affected by anti-androgen treatment. The aim of this article is to briefly review the most relevant clinical features of three main genital markers of the male dog.

  19. Biochemical and hormonal analysis of follicular fluid and serum of female dromedary camels (Camelus dromedarius) with different sized ovarian follicles.

    El-Bahr, S M; Ghoneim, I M; Waheed, M M


    The current study aimed to compare some biochemical and hormonal constituents in follicular fluids and serum of female dromedary camels with different sized ovarian follicles. Therefore, follicular fluids from follicles sized 1.1-1.5cm (n=10), 1.6-2.1cm (n=10) and 2.2-2.5cm (n=10) and sera were harvested from 20 female camels. The concentrations of ascorbic acid, glucose, cholesterol and activities of acid phosphatase (ACP) and alkaline phosphatase (ALP) were not changed significantly neither in follicular fluids of all follicle sizes nor in sera of female camels with different sized follicles. The concentrations of estradiol-17β (E2) in the follicular fluid of follicles sized 2.2-2.5cm were significantly lower (P<0.01) than its corresponding value in follicular fluid of other follicle sizes. The concentrations of progesterone (P4), tri-iodothyronine (T3), thyroxin (T4), cortisol and insulin-like growth factor-1 (IGF-1) remained comparable in follicular fluids of all examined different sized follicles. The concentrations of E2, P4, T3, T4, cortisol and IGF-1 were similar in the serum of camels with different sized follicles. Interestingly, mean concentrations of P4 and IGF-1 in follicular fluids were higher than their corresponding values in sera of camels with different sized follicles and the mean concentrations of glucose, cholesterol, ALP and cortisol in sera were higher than their corresponding values in follicular fluids of the examined camels. With the exception of E2, there were no significant differences in biochemical and hormonal constituents between follicular fluids from different sized follicles.

  20. Defining Starch Binding by Glucan Phosphatases

    Auger, Kyle; Raththagala, Madushi; Wilkens, Casper;


    Starch is a vital energy molecule in plants that has a wide variety of uses in industry, such as feedstock for biomaterial processing and biofuel production. Plants employ a three enzyme cyclic process utilizing kinases, amylases, and phosphatases to degrade starch in a diurnal manner. Starch...... is comprised of the branched glucan amylopectin and the more linear glucan amylose. Our lab has determined the first structures of these glucan phosphatases and we have defined their enzymatic action. Despite this progress, we lacked a means to quickly and efficiently quantify starch binding to glucan...... phosphatases. The main objective of this study was to quantify the binding affinity of different enzymes that are involved in this cyclic process. We established a protocol to quickly, reproducibly, and quantitatively measure the binding of the enzymes to glucans utilizing Affinity Gel Electrophoresis (AGE...

  1. Defining Starch Binding by Glucan Phosphatases

    Auger, Kyle; Raththagala, Madushi; Wilkens, Casper


    phosphatases. The main objective of this study was to quantify the binding affinity of different enzymes that are involved in this cyclic process. We established a protocol to quickly, reproducibly, and quantitatively measure the binding of the enzymes to glucans utilizing Affinity Gel Electrophoresis (AGE...... glucan phosphatases showed similar affinities for the short oligosaccharide β-cyclodextrin. We performed structure-guided mutagenesis to define the mechanism of these differences. We found that the carbohydrate binding module (CBM) domain provided a stronger binding affinity compared to surface binding...

  2. Titania nanotube delivery fetal bovine serum for enhancing MC3T3-E1 activity and osteogenic gene expression

    Peng, Jing, E-mail: [Airport College, Civil Aviation University of China, Tianjin 300300 (China); Zhang, Xinming, E-mail: [Tianjin Product Quality Inspection Technology Research Institute, Tianjin 300384 (China); School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China); Li, Zhaoyang [School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China); Liu, Yunde [School of Medical Laboratory, Tianjin Medical University, Tianjin 300203 (China); Yang, Xianjin [School of Materials Science and Engineering, Tianjin University, Tianjin 300072 (China)


    Titania nanotube (TNT) delivery of fetal bovine serum (FBS) was conducted on titanium (Ti) to enhance bone tissue repair. Scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS) showed FBS increased the tube wall thickness and decreased the tube diameter. Attenuated total reflectance Fourier transform infrared further confirmed that FBS completely covered the TNT and changed the surface composition. Water contact angle tests showed TNT/FBS possessed hydrophilic properties. Compared to original Ti, the TNT/FBS group had more attached osteoblasts after 2 h and enhanced filopodia growth at 0.5 h. Significantly, more osteoblasts were also observed on TNT/FBS after 7 d culturing. FBS was released steadily from TNT; about 70% of FBS had been released at 3 d and 90% at 5 d, as shown by the bicinchoninic acid method. TNT/FBS also enhanced subsequent osteoblast differentiation and gene expression; the quantum real-time polymerase chain reaction test showed that TNT/FBS up-regulated alkaline phosphatase and osteocalcin gene expression at 7 d and 14 d. Therefore, TNT/FBS delivered sustained in situ nutrition and enhanced osteoblast activity and osteogenic gene expression. - Highlights: • Fetal Bovine Serum (FBS) was filled in titania nanotube (TNT) structures. • FBS provided sustained-release in situ nutrition for surface osteoblast growth. • TNT/FBS enhanced osteoblast activity and osteogenic gene expression.

  3. Effects of Vitamin D Supplementation on Serum 25-Hydroxyvitamin D Concentrations in Cirrhotic Patients: A Randomized Controlled Trial

    Stefan Pilz


    Full Text Available Background: The liver is crucial for 25-hydroxyvitamin D (25(OHD metabolism, and vitamin D deficiency is highly prevalent in patients with cirrhosis and predicts adverse outcomes. We aimed to evaluate whether vitamin D supplementation in patients with cirrhosis is effective in increasing 25(OHD serum concentrations. Secondary outcome measures included liver function tests (aspartate aminotransferase (AST, alanine aminotransferase (ALT, gamma glutamyltransferase (GGT, and alkaline phosphatase (AP, albumin, International Normalized Ratio (INR, bilirubin, the liver fibrosis marker hyaluronic acid, and parameters of mineral metabolism including parathyroid hormone (PTH. Methods: This is a double-center, double-blind, placebo-controlled study conducted from December 2013 to May 2014 at the Medical University of Graz, and the hospital Hoergas-Enzenbach, Austria. We enrolled 36 consecutive patients with cirrhosis and 25(OHD concentrations below 30 ng/mL. Study participants were randomly allocated to receive either 2800 International Units of vitamin D3 per day as oily drops (n = 18 or placebo (n = 18 for 8 weeks. Results: Thirty-three study participants (mean (SD age: 60 (9 years; 21% females; 25(OHD: 15.6 (7.4 ng/mL completed the trial. The mean treatment effect (95% CI for 25(OHD was 15.2 (8.0 to 22.4 ng/mL (p < 0.001. There was no significant effect on any secondary outcome. Conclusions: In this randomized controlled trial, vitamin D supplementation increases 25(OHD serum concentrations, even in cirrhotic patients.

  4. Diagnosis value of serum ALP and osteocalcin in early prostate cancer bone metastasis

    Zong-Yong Cheng


    Objective:To explore the role and significance of the joint detection of serum alkaline phosphatase (ALP) and osteocalcin in prostate cancer bone metastases.Methods:A total of 87 cases of prostate cancer patients were diagnosed by radionuclide bone imaging, and 51 cases of them were included in the bone metastases group, while the other 36 cases were selected as the non-metastases group. Serum levels of ALP and osteocalcin of all patients were detected. The sensitivity, specificity, accuracy, likelihood ratio and the predictive value of patients in the two groups were analyzed. The sensitivity and specificity of the combined detection of ALP and osteocalcin and their expression levels in different bone metastases degrees were analyzed.Results:Serum ALP and osteocalcin levels of patients in metastases group were higher than those in non-metastases group and normal control group. In non-metastases group, the ALP level was higher than that in normal control group, while its osteocalcin level was lower than that in control group (P<0.05); The sensitivities of ALP and osteocalcin were 77.2% and 70.6%, respectively, and their specificities were 61.1% and 54.6%, respectively. The sensitivity and specificity became 93.3% and 82.33% in combined detection of ALP and osteocalcin, which was significantly higher than the single detection (P<0.05). The expression levels of ALP and osteocalcin increased with the increase of the metastases degrees (P<0.05). Conclusions:Combined detection of ALP and osteocalcin can be used in the early diagnosis of prostate cancer with improved diagnostic efficiency.

  5. Correlation of serum parathyroid hormone with mineral bone disease in chronic kidney disease patients

    Rajeshwari S Vhora


    Full Text Available Background: Mineral bone disease (MBD is a systemic disorder of mineral and bone metabolism due to chronic kidney disease (CKD. Bone disease in CKD is due to secondary hyperparathyroidism. Serum intact parathyroid hormone (iPTH level estimation is a potential noninvasive method for the diagnosis of MBD at early stage. Aim: Treating renal bone disease should be one of the primary aims of therapy for CKD. Evaluation of the biochemical parameters of CKD-MBD (primarily phosphorus, calcium, parathyroid hormone, and Vitamin D levels as early as CKD stage 3, and an assessment of bone status (by the best means available, should be used to guide treatment decisions. The adverse effects of high phosphorus intake relative to renal clearance (including stimulation of hyperparathyroidism precede hyperphosphatemia, which presents late in CKD. Early reduction of phosphorus load may ameliorate these adverse effects. Evidence that calcium load may influence progression of vascular calcification with effects on mortality, should also be considered when choosing the type and dose of phosphate binder to be used. MBD in CKD has high morbidity and mortality and hence it is important to detect it at an early stage. iPTH levels can be highly sensitive and it is one of the useful noninvasive biochemical parameters to detect MBD in CKD. Materials and Methods: This was an observational study carried out in a tertiary care teaching hospital. The study involved 60 patients of CKD. Detailed history, physical examination, and biochemical parameters were assessed in all of them. Results: There was a significant association between hypertension, diabetes with nephropathy, and highly significant association between serum iPTH and raised blood urea levels in MBD group, however there was no significant association between duration of CKD, hemoglobin, creatinine, uric acid, phosphorous, calcium, and alkaline phosphatase with MBD. Conclusions: MBD in CKD can be detected at early

  6. 2nd Generation Alkaline Electrolysis

    Yde, Lars; Kjartansdóttir, Cecilia Kristin; Allebrod, Frank;

    This report provides the results of the 2nd Generation Alkaline Electrolysis project which was initiated in 2008. The project has been conducted from 2009-2012 by a consortium comprising Århus University Business and Social Science – Centre for Energy Technologies (CET (former HIRC)), Technical...

  7. Protein-tyrosine phosphatases in zebrafish gastrulation

    van Eekelen, M.J.L.


    Protein tyrosine phosphorylation plays a key role in relaying external stimuli and signals into the cell towards the appropriate responses. This process is mediated by protein-tyrosine kinases adding a phosphor group to a tyrosine residue and protein-tyrosine phosphatases removing a phosphor group f

  8. Enzyme kinetic characterization of protein tyrosine phosphatases

    Peters, Günther H.J.; Branner, S.; Møller, K. B.


    Protein tyrosine phosphatases (PTPs) play a central role in cellular signaling processes, resulting in an increased interest in modulating the activities of PTPs. We therefore decided to undertake a detailed enzyme kinetic evaluation of various transmembrane and cytosolic PTPs (PTPalpha, PTPbeta...

  9. Alkaline resistant ceramics; Alkalimotstaandskraftiga keramer

    Westberg, Stig-Bjoern [Vattenfall Utveckling AB, Aelvkarleby (Sweden)


    Despite durability in several environments, ceramics and refractories can not endure alkaline environments at high temperature. An example of such an environment is when burning biofuel in modern heat and power plants in which the demand for increasing efficiency results in higher combustion temperatures and content of alkaline substances in the flue gas. Some experiences of these environments has been gained from such vastly different equipment as regenerator chambers in the glass industry and MHD-generators. The grains of a ceramic material are usually bonded together by a glassy phase which despite it frequently being a minor constituent render the materials properties and limits its use at elevated temperature. The damage is usually caused by alkaline containing low-melting phases and the decrease of the viscosity of the bonding glass phase which is caused by the alkaline. The surfaces which are exposed to the flue gas in a modern power plant are not only exposed to the high temperature but also a corroding and eroding, particle containing, gas flow of high velocity. The use of conventional refractory products is limited to 1300-1350 deg C. Higher strength and fracture toughness as well as durability against gases, slag and melts at temperatures exceeding 1700 deg C are expected of the materials of the future. Continuous transport of corrosive compounds to the surface and corrosion products from the surface as well as a suitable environment for the corrosion to occur in are prerequisites for extensive corrosion to come about. The highest corrosion rate is therefore found in a temperature interval between the dew point and the melting point of the alkaline-constituent containing compound. It is therefore important that the corrosion resistance is sufficient in the environment in which alkaline containing melts or slag may appear. In environments such as these, even under normal circumstances durable ceramics, such as alumina and silicon carbide, are attacked

  10. Impacts of Persian Gulf blackfin stonefish crude venom on the haematological factors and serum enzymes levels of laboratory rat

    Amir Vazirizadeh


    Full Text Available Background: One of the venomous fishes of Persian Gulf is a type of stonefish called blackfin stonefish (Pseudosynanceia melanostigma that is very poorly-known. This fish lives on the muddy bottoms of marine shallow waters and have venomous spines on its stone like body. The envenomation usually is happened by unwanted contact of human body to its body and the proteinic venom is injected into the skin. Upon the venom entrance to the body various symptoms and signs will occur that their intensity depends on the venom amount.The aim of study is the understanding of some pharmacological impacts of blackfin stonefish for future studies. Material and Methods: A total of 18 male laboratory rats were divided into three groups of control, second and third and envenomed by sub lethal doses (1/3 LD50 and ½ LD50 intravenously and the serum enzymes namely Creatine Phosphokinase, Lactate Dehydrogenase, Glutamate-Oxaloacetate Transaminase, Gamma-Glutamyl Transpeptidase , Alkaline Phosphatase and Amylase, serum electrolytes namely Sodium. Potassium and Calcium and also complete blood cells of control and experimental rats were measured. Results: The serum enzymes levels, potassium and calcium levels and withe blood cells count in envenomed rats by blackfin stonefish crude venom in compare with control rats had significant increase while the haemoglobin level, red blood cells count and also serum sodium level had significant decrease (p<0.001. Conclusion: The increase in hepatomascular enzyme levels, and the decrease in haemoglobin level can be a probable marker of the presence of rhabdomyolysis, haemolysis and also hepatotoxicity activities in the blackfin stonefish venom and that needs to the histopathologic studies in these systems.

  11. Prognostic significance of pretreatment serum levels of albumin, LDH and total bilirubin in patients with non-metastatic breast cancer.

    Liu, Xiaoan; Meng, Qing H; Ye, Yuanqing; Hildebrandt, Michelle A T; Gu, Jian; Wu, Xifeng


    Liver function tests (LFTs) have been reported as independent predictors of non-liver disease-related morbidity and mortality in general population and cancer patients. In this study, we evaluated the relationship between pretreatment serum LFTs and overall survival (OS) in non-metastatic Caucasian breast cancer patients. Seven LFTs, including albumin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase (LDH), total bilirubin and total protein, were measured in pretreatment serum from 2425 female Caucasian patients with newly diagnosed, histologically confirmed non-metastatic invasive breast cancer. Multivariate Cox model was used to estimate hazard ratio (HR) and 95% confidence interval (CI) for the association of individual LFTs with 5-year OS while adjusting for age, smoking status, pathological characteristics and treatment regimen. We found that serum albumin, LDH and total bilirubin were significantly associated with 5-year OS in multivariate Cox analyses. Patients with higher albumin level exhibited 45% reduced risk of death (HR = 0.55, 95% CI: 0.40-0.75) compared with those with lower albumin level. Patients with higher total bilirubin level had a nearly 40% reduction in the risk of death (HR = 0.62, 95% CI: 0.45-0.85) and patients with higher LDH levels had a 1.42-fold increased risk of death (HR = 1.42, 95% CI: 1.08-1.88). Furthermore, cumulative analysis showed a significant dose-response trend of significantly increasing risk of death with increasing number of unfavorable LFT levels. Our result highlighted the potential of using pretreatment serum levels of albumin, LDH and total bilirubin as prognostic factors for OS in patients with non-metastatic breast cancer.

  12. Serum sickness

    ... the problem should be stopped. Avoid using that medicine or antiserum in the future. ... Call your provider if you received medicine or antiserum in the last 4 weeks and have symptoms of serum sickness.

  13. Influence of physical and emotional activity on the metabolic profile of blood serum of race horses

    T. I. Bayeva


    Full Text Available In the article data are presented on dynamics of the level of indicators of metabolic profile of blood serum of race horses of the Ukrainian riding breed in the conditions of physical and emotional loading. Clinically healthy race horses were the object of  research. Blood was taken from the jugular vein to obtain serum and for further biochemical research. For the research 12 race horses from a training group were chosen. From time to time the animals took part in competitions; they were not specially used in races and were mostly used for the training of junior riders and sportsmen of different levels. Blood was taken in conditions of relative rest after ordinary training and after emotional stress during the entertainment performances when a large number of people were present and loud music was played. In the blood serum the following biochemical indicators were defined: whole protein, urea, creatinine, uric acid, total bilirubin and its fractions, glucose, cholestererol, triacylglycerol, calcium, ferrum, lactate, pyruvate, activity of the AlAT, SGOT, GGTP, LDH, an alkaline phosphatase – which makes it possible to determine reasonably accurately the adaptation potential of a horse under various types of loading. We established that during training and psychoemotional loading of racing horses of the training group of the Ukrainian riding breed, multidirectional changes in the level of biochemical indicators of blood serum occurred, which is evidence of stress in the metabolic processes in the animals’ organisms. Concentration of a biomarker of an oxidative stress, uric acid, increased after physical loading by 8.6%, and after emotional loading by 55.1%, which demonstrates that emotional stress had the more negative effect, indicating insufficient adaptation by the horses before demonstration performances. After physical loading, reaction of transamination in the horses’ liver cells intensified, and after emotional loading its intensity

  14. Short-term bisphosphonate treatment reduces serum 25(OH vitamin D3 and alters values of parathyroid hormone, pentosidine, and bone metabolic markers

    Kamimura M


    Full Text Available Mikio Kamimura,1 Shigeharu Uchiyama,2 Yukio Nakamura,2,3 Shota Ikegami,2 Keijiro Mukaiyama,2 Hiroyuki Kato2 1Center for Osteoporosis and Spinal Disorders, Kamimura Orthopaedic Clinic, Matsumoto, Japan; 2Department of Orthopaedic Surgery, Shinshu University School of Medicine, Matsumoto, Japan; 3Department of Orthopedic Surgery, Showa-Inan General Hospital, Komagane, Japan Abstract: This study aimed to clarify the effects of short-term bisphosphonate (BP administration in Japanese osteoporotic patients retrospectively. Daily minodronate (MIN at 1 mg/day (MIN group or weekly risedronate (RIS at 17.5 mg/week (RIS group was primarily prescribed for each patient. We analyzed the laboratory data of 35 cases (18 of MIN and 17 of RIS before the start of treatment and at 4 months afterward. The changes in 25(OHD3, whole parathyroid hormone (PTH, serum pentosidine, and the bone turnover markers urinary cross-linked N-telopeptide of type I collagen (NTX, serum tartrate-resistant acid phosphatase (TRACP-5b, bone-specific alkaline phosphatase (BAP, and undercarboxylated osteocalcin were evaluated. Overall, serum 25(OHD3 was significantly decreased from 21.8 to 18.4 ng/mL at 4 months, with a percent change of –14.7%. Whole PTH increased significantly from 23.4 to 30.0 pg/mL, with a percent change of 32.1%. Serum pentosidine rose from 0.0306 to 0.0337 µg/mL, with a percent change of 15.2%. In group comparisons, 25(OHD3 and pentosidine showed comparable changes in both groups after 4 months of treatment, whereas whole PTH became significantly more increased in the MIN group. All bone turnover markers were significantly decreased at 4 months in both groups. Compared with the RIS group, the MIN group exhibited significantly larger value changes for urinary NTX, serum TRACP-5b, and BAP at the study end point. This study demonstrated that serum 25(OHD3 became significantly decreased after only 4 months of BP treatment in Japanese osteoporotic patients and

  15. [Evaluation of serum PIVKA-II by Lumipulse PrestoII assay].

    Hiramatsu, Kumiko; Tanaka, Yasuhito; Takagi, Kazumi; Kani, Satomi; Goto, Takaaki; Takasaka, Yoshimitsu; Matsuura, Kentaro; Sugauchi, Fuminaka; Moriyama, Kazushige; Murakami, Hiroshi; Kitajima, Sachiko; Mizokami, Masashi


    Measurements of serum concentrations of Des-gamma-carboxy Prothrombin (PIVKA-II) are widely used for diagnosing hepatocellular carcinoma (HCC). Recently, in Lumipulsef assay, it was reported that antibodies against alkaline phosphatase (ALP) derived from anti bleeding sheets led false high values of PIVKA-II in the patients with HCC resection. To improve the previous issue, newly developed Lumipulse PrestoII assay was examined. (1) The assay was reliable and positively correlated with the previous assays (Lumipulse f and Picolumi, R = 0.997 and 0.994 (n=115), respectively). (2) Eleven cases, which had false high values of PIVKA-II by the Lumipulsef assay, were examined by the PrestoII assay with excess of inactive ALP. The false high values of 10 cases were improved, but only one was still high. False reactivity of this case was stronger than other cases, more effective adsorption was required. (3) Comparing the absorbent activity of inactive ALP among 6 different kinds, we found inactive ALP with much higher adsorbent activity. When this inactive ALP was applied to assay, false high values of PIVKA-II were improved in all 11 cases. In conclusion, the PrestoII assay, which applies the inactive ALP with high activity, is reliable and useful for clinical screening.

  16. Development of alkaline fuel cells.

    Hibbs, Michael R.; Jenkins, Janelle E.; Alam, Todd Michael; Janarthanan, Rajeswari; Horan, James L.; Caire, Benjamin R.; Ziegler, Zachary C.; Herring, Andrew M.; Yang, Yuan; Zuo, Xiaobing; Robson, Michael H.; Artyushkova, Kateryna; Patterson, Wendy; Atanassov, Plamen Borissov


    This project focuses on the development and demonstration of anion exchange membrane (AEM) fuel cells for portable power applications. Novel polymeric anion exchange membranes and ionomers with high chemical stabilities were prepared characterized by researchers at Sandia National Laboratories. Durable, non-precious metal catalysts were prepared by Dr. Plamen Atanassovs research group at the University of New Mexico by utilizing an aerosol-based process to prepare templated nano-structures. Dr. Andy Herrings group at the Colorado School of Mines combined all of these materials to fabricate and test membrane electrode assemblies for single cell testing in a methanol-fueled alkaline system. The highest power density achieved in this study was 54 mW/cm2 which was 90% of the project target and the highest reported power density for a direct methanol alkaline fuel cell.

  17. Assessment and kinetics of soil phosphatase in Brazilian Savanna systems



    Full Text Available The activity and kinetics of soil phosphatases are important indicators to evaluate soil quality in specific sites such as the Cerrado (Brazilian Savanna. This study aimed to determine the activity and kinetic parameters of soil phosphatase in Cerrado systems. Soil phosphatase activity was assessed in samples of native Cerrado (NC, no-tillage (NT, conventional tillage (CT and pasture with Brachiaria brizantha (PBb and evaluated with acetate buffer (AB, tris-HCl buffer (TB, modified universal buffer (MUB and low MUB. The Michaelis-Menten equation and Eadie-Hofstee model were applied to obtain the kinetic parameters of soil phosphatase using different concentrations of p-nitrophenol phosphate (p-NPP. MUB showed the lowest soil phosphatase activity in all soils whereas AB in NC and NT presented the highest. Low MUB decreased interferences in the assessment of soil phosphatase activity when compared to MUB, suggesting that organic acids interfere on the soil phosphatase activity. In NC and NT, soil phosphatase activity performed with TB was similar to AB and low MUB. Km values from the Michaels-Menten equation were higher in NC than in NT, which indicate a lower affinity of phosphatase activity for the substrate in NC. Vmax values were also higher in NC than in NT. The Eadie-Hofstee model suggests that NC had more phosphatase isoforms than NT. The study showed that buffer type is of fundamental importance when assessing soil phosphatase activity in Cerrado soils.

  18. The effects of aqueous extract of stevia plant (Stevia rebaudiana on serum concentration of vaspin and Angiopoietin-like Protein-3 in streptozotocin induced diabetic rats

    Samad Akbarzadeh


    Full Text Available Background: Considering the importance of the prevalence of diabetes which involves approximately 6% of the world's adult population, the need for low-calorie natural sweetener is felt more than ever. Recent studies have shown that hormones such as vaspin and Angiopoietin-like Protein3 (ANGPTL3 are associated with diabetes. The purpose of this study is to evaluate the effect of Stevia (Stevia rebaudiana plant (as a low calorie sugar on serum concentration of vaspin and ANGPTL3 in diabetic rats. Materials and Methods: In this case-control study, forty male wistar rats weighing 180-250 g were divided into 5 equal groups: control, diabetic control and doses of 250, 500 and 750 mg/kg/BW/day of Stevia extract treatment. Diabetes was induced by intravenous injection of Streptozotocin (60 mg/kg. After 5 days, the rats with glucose above 300 mg/dl were considered as diabetic. The Stevia treatment groups received 250, 500 and 750 mg of Stevia extract for thirty days. At the end of experiment, blood samples were obtained measurement of vaspin, ANGPTL3, triglycerides, cholesterol, HDL, glucose, insulin and ALP. Histological study of the pancreas and liver biopsy were also performed. The results of the treatment and control groups were analyzed by SPSS software and P<0.05 was considered statistically significant. Results: The level of alkaline phosphatase, insulin resistance index, glucose and triglyceride were decreased significantly in the groups 250 and 500 compared with the diabetic control group(p≤0.05. However, insulin levels, HOMA.B, vaspin, ANGPTL3 and weight of the rats in all treatment groups were not significantly different from the control diabetic group. There were no histological changes in pancreatic and liver tissue following Stevia treatment. Conclusion: Administration of Stevia extract via reduction in serum glucose, triglyceride and insulin resistance can be effective in lowering the blood sugar and lipid, also lowering concentrations

  19. Acid Phosphatase Development during Ripening of Avocado.

    Sacher, J A


    The activity and subcellular distribution of acid phosphatase were assayed during ethylene-induced ripening of whole fruit or thick slices of avocado (Persea americana Mill. var. Fuerte and Hass). The activity increased up to 30-fold during ripening in both the supernatant fraction and the Triton X-100 extract of the precipitate of a 30,000g centrifugation of tissue homogenates from whole fruit or slices ripening in moist air. Enzyme activity in the residual precipitate after Triton extraction remained constant. The development of acid phosphatase in thick slices ripened in moist air was similar to that in intact fruit, except that enzyme development and ripening were accelerated about 24 hours in the slices. The increase in enzyme activity that occurs in slices ripening in moist air was inhibited when tissue sections were infiltrated with solutions, by aspiration for 2 minutes or by soaking for 2 hours, anytime 22 hours or more after addition of ethylene. This inhibition was independent of the presence or absence of cycloheximide or sucrose (0.3-0.5m). However, the large decline in enzyme activity in the presence of cycloheximide, as compared with the controls, indicated that synthesis of acid phosphatase was occurring at all stages of ripening.

  20. Relationship between serum leptin levels and bone mineral density and bone metabolic markers in patients on hemodialysis

    Farokhlagha Ahmadi


    Full Text Available Leptin is the protein product of the obesity gene, which is produced in fat tissue. It was originally thought to be involved only in the regulation of food intake and energy balance. We aimed to investigate the relationship of serum leptin levels with bone mineral density (BMD and biochemical markers of bone turnover in patients on hemodialysis (HD. This study included 72 patients (43 males and 29 females, whose mean age was 55.1 ± 11.4 years, mean body mass index was 23.13 ± 2.75 kg/m 2 and mean duration on HD was 5 ± 3.4 years. The BMD values were calculated using dual-energy X-ray absorptiometry (DEXA at the femoral neck and lumbar spine. Blood samples were taken for leptin, intact parathyroid hormone (I-PTH, bone alkaline phosphatase (BAP, calcium (Ca, phosphate (P and albumin. The leptin levels were higher in females than in males (22.3 ± 19.6 vs 20.8 ± 23, but this difference was not significant. The serum leptin level had a strong positive correlation with Ca levels in the female patients (r = 0.659 and P = 0.01 and a negative correlation with albumin levels (r = -0.461 and P = 0.01. No correlation was found with age, BMI, duration on dialysis, BMD and serum levels of PTH, BAP and P for the entire patient group or either gender separately. The serum leptin level was significantly lower in females with PTH >300 pg/mL when compared with patients with PTH = 100-300 pg/mL (86 ± 85 vs 47 ± 48 (P = 0.011.Women with BAP <300 IU/L had significantly higher serum leptin than those with BAP 300-600 IU/L (P = 0.024. Women with Ca <8.5 mg/dL had significantly lower serum leptin levels compared with those with Ca levels of 8.5-10.5 mg/dL (P = 0.011. There was no significant difference between the two genders among variables such as age, BMI, duration on dialysis, serum leptin, I-PTH, Ca, P, BAP, albumin and BMD of the femoral neck and lumbar spine.

  1. Primary structure of rat secretory acid phosphatase and comparison to other acid phosphatases.

    Roiko, K; Jänne, O A; Vihko, P


    Overlapping cDNA clones encoding rat prostatic acid phosphatase (rPAP) were isolated by using two human prostatic acid phosphatase (hPAP)-encoding cDNAs to screen rat prostatic cDNA libraries. The isolated cDNAs encompassed a total of 1626 nucleotides (nt), of which 1143 nt corresponded to the protein coding sequence encoding a mature polypeptide of 350 amino acids (aa) and a 31-aa long signal peptide-like sequence. The deduced Mr of the mature rPAP was 40,599. RNA blot analysis indicated the presence of three mRNA species (4.9, 2.3 and 1.5 kb in size) in the rat prostate. The deduced aa sequences of rPAP and hPAP show 75% identity, whereas the similarity between rPAP and human lysosomal acid phosphatase (hLAP) is only 45%. Furthermore, the sequence similarity between rPAP and rat lysosomal acid phosphatase (rLAP) is 46% at the aa level. Similar to hPAP, but unlike hLAP and rLAP, the rPAP sequence lacks a membrane-anchoring domain indicating the secretory character of this phosphatase. All six cysteines present in the overlapping areas of the mature rPAP, hPAP, rLAP and hLAP proteins are positionally conserved, suggesting that these residues are important for the tertiary structure of acid phosphatases (APs). The previously reported active site residues, two arginines and one histidine, are also conserved in these APs.

  2. Age- and sex-related reference values for serum insulin concentration and its biological determinants in a French healthy population. The STANISLAS cohort.

    François, Aurélie; Maumus, Sandy; Vincent-Viry, Monique; Guéguen, René; Siest, Gérard; Visvikis, Sophie


    Insulin is involved in coronary heart disease through diabetes and metabolic syndrome. A great deal is known about insulin and its correlates, as well as factors related to changes in insulin. However, few studies consider the broad variety of correlates simultaneously. Therefore, the aims of the present study were to characterize the main factors of biological variation affecting serum insulin concentration and to establish reference limits of insulinemia in a presumably healthy French population. Insulin was measured using a microparticular enzymatic immunoassay. A total of 646 subjects aged 11-58 years from the STANISLAS cohort and divided into four groups of 162 males, 157 females, 163 boys and 164 girls, were included in the statistical analyses. In the whole population, serum insulin concentration varied from 0.80 to 54.60 microU/ml. Significant factors affecting insulin were age, gender, body mass index and glucose, in addition to alanine aminotransferase and high-density lipoprotein cholesterol in men, triglycerides and oral contraceptive use in women, and alkaline phosphatase in girls. In summary, we presented biological correlates of insulin in both healthy French male and female adults and children/adolescents and determined reference limits for insulin for each group. These results will contribute to a better interpretation of insulin data in further studies and laboratory investigations.

  3. A retrospective study of the clinical hematology and the serum biochemistry tests made on canine dirofilariasis cases in an animal hospital population in Bangkok, Thailand.

    Niwetpathomwat, Anuchai; Kaewthamasorn, Morakot; Tiawsirisup, Sonthaya; Techangamsuwan, Somporn; Suvarnvibhaja, Siram


    Heartworm disease, caused by the filarial nematode, Dirofilaria immitis, is a major, potentially life-threatening disease of dogs, with worldwide distribution and global significance. It is not only of veterinary importance but it also has zoonotic potential in many regions. It is considered as an endemic disease in Thailand, although clinical data about the disease is rarely reported. The objectives of this study were to characterize the clinical hematology and the biochemistry of canine dirofilariasis cases admitted to Chulalongkorn University, Small Animal Teaching Hospital in Bangkok, Thailand, from 2001-2003. All hematology and serum biochemistry parameter interpretations were based on reference values. A total of 1023 dogs were evaluated in this study. Dogs were divided into three groups based on their heartworm classification (microfilaremic, occult and negative). The major hematological findings in microfilaremic dogs were a mild to moderate anemia, mild to severe thrombocytopenia, marked leukocytosis, moderate to marked neutrophilia, eosinophilia and monocytosis. The most common serum biochemical abnormalities in microfilaremic dogs included increased alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase with a mean+/-SD of 311+/-299, 82+/-76 and 50+/-38 IU/L, respectively. It is likely that this parasitic infection led to the impairment of the hematological and biochemical status of the infected dogs. We believe that investigation of these laboratory based parameters, when associated with the clinical signs, is a very important approach to be considered in the routine clinical follow up, as well as being important for therapeutic evaluations.

  4. Effect of calcitriol, cinacalcet combined with nursing intervention on patients with MHD SHPT serum fibroblast growth factor 23

    Le Chen; Xiao-Yun Wu


    Objective:To investigate effect of calcitriol, cinacalcet combined with nursing intervention on maintenance hemodialysis (MHD) patients with secondary hyperparathyroidism (SHPT) serum fibroblast growth factor 23 (FGF23) effect.Methods:A total of 90 cases of SHPT in our hospital from March 2016 to March 2014 were selected and randomly divided into the observation group and the control group. The 2 groups were given nursing intervention on cinacalcet hydrochloride swallow when eating, starting dose is 25 mg/d, every 2 to 4 weeks according to the parathyroid hormone (iPTH), calcium (Ca), phosphorus (P) level adjustment dose, the maximum dose of not more than 75 mg/d, the observation group was treated with basic treatment in the Calcitriol Soft Capsules 0.25 g/d, 3 times/week, evaluate the effect of the 2 groups after 3 months of treatment. The 2 groups before and after treatment collected fasting peripheral venous blood, Ca, P were measured by colorimetric method, determination of iPTH and FGF23 in the serum and bone specific alkaline phosphatase ELISA (BSAP), serum creatinine automatic biochemistry analyzer (Scr), nitrogen (BUN), uremic quantitative detection and analysis of urea clearance index of single room urea kinetic model (KT/V). Results:Level of Ca, P of observation group after treatment increased and iPTH decreased more significantly compared with that of control group (P0.05), FGF23 and BSAP levels after treatment in observation group decreased more significantly than the control group (P<0.05). Conclusions: Calcitriol, cinacalcet combined with nursing intervention in the treatment of MHD patients with SHPT, can effectively reduce the PTH, FGF23, BSAP level, improve the clinical symptoms.

  5. Alkaline Water and Longevity: A Murine Study.

    Magro, Massimiliano; Corain, Livio; Ferro, Silvia; Baratella, Davide; Bonaiuto, Emanuela; Terzo, Milo; Corraducci, Vittorino; Salmaso, Luigi; Vianello, Fabio


    The biological effect of alkaline water consumption is object of controversy. The present paper presents a 3-year survival study on a population of 150 mice, and the data were analyzed with accelerated failure time (AFT) model. Starting from the second year of life, nonparametric survival plots suggest that mice watered with alkaline water showed a better survival than control mice. Interestingly, statistical analysis revealed that alkaline water provides higher longevity in terms of "deceleration aging factor" as it increases the survival functions when compared with control group; namely, animals belonging to the population treated with alkaline water resulted in a longer lifespan. Histological examination of mice kidneys, intestine, heart, liver, and brain revealed that no significant differences emerged among the three groups indicating that no specific pathology resulted correlated with the consumption of alkaline water. These results provide an informative and quantitative summary of survival data as a function of watering with alkaline water of long-lived mouse models.

  6. Interaction of alkali and alkaline earth ions with Ochratoxin A

    Poor, Miklos [Institute of Laboratory Medicine, University of Pecs, Pecs H-7624 (Hungary); Kunsagi-Mate, Sandor; Matisz, Gergely; Li, Yin; Czibulya, Zsuzsanna [Department of General and Physical Chemistry, University of Pecs, Pecs H-7624 (Hungary); Janos Szentagothai Research Center, Pecs H-7624 (Hungary); Peles-Lemli, Beata [Department of General and Physical Chemistry, University of Pecs, Pecs H-7624 (Hungary); Koszegi, Tamas, E-mail: [Institute of Laboratory Medicine, University of Pecs, Pecs H-7624 (Hungary)


    The effect of alkali and alkaline earth ions on the chemical equilibrium of mono- and dianionic forms of the mycotoxin Ochratoxin A (OTA) and their bonding onto the surface of Bovine Serum Albumin (BSA) have been investigated by fluorescence spectroscopy and fluorescence polarization techniques. Our results show that alkali metal ions shift the chemical equilibrium towards formation of dianionic form of OTA. Furthermore, the alkaline earth ions can compete with BSA for binding to OTA when these ions are present in millimolar concentrations. Our data also highlight the possibility that the 'free' fraction of OTA (not bound onto the surface of albumin) or at least a part of it is present in cation-bound form in body fluids. These observations are supported by stability constants and quantum-chemical calculations. Among the studied alkaline metal ions magnesium showed the highest affinity towards OTA under physiological conditions. Further research is required to analyze the potential significance of Mg{sup 2+}-OTA complex in cellular uptake and/or elimination of the toxin in the human body. - Highlights: Black-Right-Pointing-Pointer Fluorescence spectroscopy reveals cation-Ochratoxin A (OTA) interactions. Black-Right-Pointing-Pointer Alkali ions shift the equilibrium of OTA to formation of a dianionic structure. Black-Right-Pointing-Pointer Alkaline earth ions directly bind to OTA in the order: Mg{sup 2+}, Ca{sup 2+}, Ba{sup 2+}. Black-Right-Pointing-Pointer Quantum chemical calculations and logK values support our experimental data.

  7. Effects of dietary supplementation of fermented Ginkgo biloba L. residues on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets.

    Zhou, Hao; Wang, Chengzhang; Ye, Jianzhong; Chen, Hongxia; Tao, Ran


    This study evaluated the effects of fermented Ginkgo biloba L. residues (FGBLR) on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets. Pigs were allotted to five dietary treatments, including negative control (NC: antibiotic free basal diet), positive control (PC) (NC + 30 mg apramycin/kg) and FGBLR-50, 100, 150 (NC + 50, 100, 150 g FGBLR/kg). Pigs in FGBLR-100 and PC treatments showed increased final body weight, average daily gain, gain:feed and apparent total tract digestibility of dry matter, N and gross energy (P < 0.05) compared with NC, FGBLR-50 and FGBLR-150 treatments, In addition, pigs fed with FGBLR-100 diet showed higher serum total protein, albumin, alkaline phosphatase, glucose, hemoglobin, total iron, total iron binding capacity, superoxide dismutase and glutathione superoxide dismutase levels, and lower serum blood urea nitrogen, malondialdehyde, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, triglyceride and total cholesterol levels than those fed with PC and NC diets (P < 0.05). Moreover, feeding FGBLR-100 could increase levels of immunoglobulin G (IgG), IgA and IgM, as well as lymphocyte transformation rates, ratio of CD4+ to CD8+ cells and proportions of CD2+, CD4+, B, major histocompatibility complex (MHC)-I and MHC-II cells, and can decrease proportion of CD8+ cells in blood of piglets compared with PC and NC groups (P < 0.05). These results indicate that dietary supplementation with 10% of FGBLR showed greatest beneficial effects on growth performance, nutrient digestibility, serum biochemical parameters and immune function in weaned piglets, which were superior to antibiotic supplemental diets.

  8. Phytase, phosphatase activity and p-nutrition of soybean as influenced by inoculation of bacillus.

    Ramesh, A; Sharma, Sushil K; Joshi, O P; Khan, I R


    The efficiency of different Bacillus isolates on rhizosphere soil enzyme activities and P-nutrition of soybean was carried out under microcosm conditions. Significant increase in enzyme activities viz., fluorescein diacetate activity, phosphatase and phytase activity and consequent effects on P-nutrition were observed with the inoculation of Bacillus isolates over uninoculated control. Among the isolates, BD-3-1B, KHBD-6, BDKH-3, Bacillus amyloliquefacians, and Bacillus cereus were found to be promising. The phytic acid-P as a percentage of total P content in soybean seeds decreased with the inoculation of Bacillus isolates as compared to un-inoculated control. A decrease in phytic-P in soybean seeds not only results in better digestibility and increased feed efficiency. Pearson correlation studies revealed a significant positive association between acid, alkaline phosphatases, phytase activity on available P content in soil and P content in seeds with the inoculation of Bacillus isolates, indicating role of these enzymes in P mobilization and acquisition by soybean.

  9. Counter-regulatory phosphatases TNAP and NPP1 temporally regulate tooth root cementogenesis

    Laura E Zweifler; Mudita K Patel; Francisco H Nociti Jr; Helen F Wimer; Jose L Milla n; Martha J Somerman; Brian L Foster


    Cementum is critical for anchoring the insertion of periodontal ligament fibers to the tooth root. Several aspects of cementogenesis remain unclear, including differences between acellular cementum and cellular cementum, and between cementum and bone. Biomineralization is regulated by the ratio of inorganic phosphate (Pi) to mineral inhibitor pyrophosphate (PPi), where local Pi and PPi concentrations are controlled by phosphatases including tissue-nonspecific alkaline phosphatase (TNAP) and ectonucleotide pyrophosphatase/phosphodiesterase 1 (NPP1). The focus of this study was to define the roles of these phosphatases in cementogenesis. TNAP was associated with earliest cementoblasts near forming acellular and cellular cementum. With loss of TNAP in the Alpl null mouse, acellular cementum was inhibited, while cellular cementum production increased, albeit as hypomineralized cementoid. In contrast, NPP1 was detected in cementoblasts after acellular cementum formation, and at low levels around cellular cementum. Loss of NPP1 in the Enpp1 null mouse increased acellular cementum, with little effect on cellular cementum. Developmental patterns were recapitulated in a mouse model for acellular cementum regeneration, with early TNAP expression and later NPP1 expression. In vitro, cementoblasts expressed Alpl gene/protein early, whereas Enpp1 gene/protein expression was significantly induced only under mineralization conditions. These patterns were confirmed in human teeth, including widespread TNAP, and NPP1 restricted to cementoblasts lining acellular cementum. These studies suggest that early TNAP expression creates a low PPi environment promoting acellular cementum initiation, while later NPP1 expression increases PPi, restricting acellular cementum apposition. Alterations in PPi have little effect on cellular cementum formation, though matrix mineralization is affected.

  10. 成纤维细胞生长因子23与血液透析患者血磷和甲状旁腺激素浓度的关系%Relationship between serum level of fibroblast growth factor 23 and phosphorus and parathyroid hormone in dialysis patients

    徐丰博; 刘惠兰; 孙懿


    目的 观察维持性血液透析患者血中成纤维细胞生长因子23(fibroblast growth factor23,FGF23)浓度,探讨维持性血液透析(maintenance hemodialysis,MHD)患者FGF23的浓度及其与血磷、甲状旁腺激素(parathyroid hormone,PTH)浓度之间的关系.方法 选取稳定MHD患者50例,健康对照30人,透析患者根据PTH浓度分为A1组(PTH500 pg/mL).应用酶联免疫分析法测定血清FGF23和1,25-二羟活性维生素D3[1,25-dihydroxy-cholecalciferol,1,25-(OH)2D3]浓度,同时测定血清PTH、血钙(Ca2+)、磷(P3-)、碱性磷酸酶(alkaline phosphatase,ALP)、白蛋白(albumin,ALB)等指标.结果①透析患者血清Log FGF23水平显著高于对照组,Log1,25-(OH)2D3水平显著低于对照组,差异有统计学意义.②Log FGF23水平在PTH>500 pg/mL组显著高于PTH 500 pg/mL). Phosphate, calcium, alkaline phosphatase in their serum were determined too. The level of FGF23 and 1 ,25-(OH)2D3 in serum were detected by the method of ELISA. Results ①The Log FGF23 in MHD group was much higher than that of control, Logl ,25-(OH)2D3 in MHD group was lower than that of control. ②The Log FGF23 in group A3 was much higher than group Al. ③Log FGF23 was positively correlated with serum phosphorus, Log PTH and Log ALP, negatively correlated with Log 1 ,25-(OH)2D3.④ Multiple linear regression analysis revealed that the level of serum level of FGF23 in MHD patients was extremely high. Conclusion The serum level of FGF23 in MHD patients was extremely high.

  11. The effect of Stevia rebaudiana on serum omentin and visfatin level in STZ-induced diabetic rats.

    Akbarzadeh, Samad; Eskandari, Fatemeh; Tangestani, Hadis; Bagherinejad, Somaieh Tangerami; Bargahi, Afshar; Bazzi, Parviz; Daneshi, Adel; Sahrapoor, Azam; O'Connor, William J; Rahbar, Ali Reza


    Recently the role of adipocytokines in relationship to incidence of diabetes has been demonstrated. One of the medicinal plants that are used in the treatment of diabetes is stevia. This study investigates the effect of stevia on serum omentin and visfatin levels as novel adipocytokines in diabetic induced rats to find potential mechanisms for the anti hyperglycemic effect of stevia. Forty male wistar rats weighing 180-250 g were induced with diabetes by intraperitoneal injection of streptozotocin (STZ). The animals were divided into 5 groups of 8. Rats in group 1 (non-diabetic control) and group 2 (diabetic control) were treated with distilled water, and the rats in the treated groups, group 3 (T250), group 4 (T500), and group 5 (T750) were treated with stevia, gavaged every day at 9 a.m. in doses of 250, 500, and 750 mg/kg, respectively. At the end of the study significant reductions in fasting blood sugar (FBS), the homeostasis model assessment insulin resistance (HOMA-IR), triglyceride (TG), alkaline phosphatase (ALP), and Omentin level were found in groups 3 and 4 in comparison with group 2. Pancreatic histopathology slides demonstrated that stevia extract did not induce any increase in the number of β-cells. The conclusion is that prescription of stevia in the doses of 250 and 500 mg/kg/d decreases the omentin level indirectly via activating insulin sensitivity and lowering blood glucose in STZ-induced diabetic rats.

  12. Superior performance of liposomes over enzymatic amplification in a high-throughput assay for myoglobin in human serum.

    Edwards, Katie A; Meyers, Katherine J; Leonard, Barbara; Baeumner, Antje J


    Myoglobin is one of several cardiac markers which become elevated in the blood following an acute myocardial infarction and can aid in the diagnosis of a heart attack. Here, a sandwich immunoassay for myoglobin was developed, including a thorough optimization of fluorescent dye-encapsulating liposomes versus enzymatic amplification (alkaline phosphatase and horseradish peroxidase) at each step. The optimized microtiter plate-based assay was capable of detecting as low as 11.3 pg/mL myoglobin and was successfully applied for the quantification of myoglobin in human serum. In comparison to enzymatic approaches, the liposomes demonstrated lower limits of detection, significantly reduced limits of quantification, improved signal discrimination through substantial signal enhancement, and reduced assay time. Liposomes were stable and functional at ambient temperatures for over 400 days. Finally, ease of use was greater due to lack of reliance on additional reagents, non-time-based signal enhancement, and excellent photostability. Optimal conditions identified for enzymatic approaches can also be used for liposome amplification, which makes substitution of these liposomes into existing assays straightforward. Thus, the extensive studies carried out here suggest that liposomes may be incorporated into formats currently utilizing enzymatic enhanced fluorescence with a potential for increased performance on various levels.

  13. A clinical evaluation of the Cobas Fara clinical chemistry analyzer for some routine serum enzymes and glucose.

    Moses, G C; Lightle, G O; Tuckerman, J F; Henderson, A R


    The authors evaluated the Cobas FARA centrifugal analyzer with respect to pipetting precision and accuracy, instrument temperature, spectrophotometric response, and analytic performance for the assay of five serum enzymes and glucose. Spectrophotometric response, temperature response, pipetting precision, and accuracy were satisfactory. However, sufficient time must be allowed for cuvet contents to reach a stable temperature before measurements are made. Total day-to-day imprecision (within plus between run) was less than 5% (coefficient of variation) for aspartate and alanine aminotransferases (AST; Enzyme Commission classification number [EC] EC; and ALT; EC; alkaline phosphatase (AP; EC; gamma-glutamyltransferase (GGT; EC; lactate dehydrogenase (LD; EC; creatine kinase (CK; EC; and glucose assays. Results compare well with those obtained with other current clinical chemistry analyzers; correlation coefficients were greater than 0.993. Sample-to-sample carryover was negligible, and method linearity was satisfactory for all tests.

  14. Multiple forms of phosphatase from human brain: isolation and partial characterization of affi-gel blue nonbinding phosphatase activities.

    Cheng, L Y; Wang, J Z; Gong, C X; Pei, J J; Zaidi, T; Grundke-Iqbal, I; Iqbal, K


    Phosphatases extracted from a human brain were resolved into two main groups, namely affi-gel blue-binding phosphatases and affi-gel blue-nonbinding phosphatases. Affi-gel blue binding phosphatases were further separated into four different phosphatase activities, designated P1-P4, and described previously. In the present study we describe the affi-gel blue-nonbinding phosphatases which were separated into seven different phosphatase activities, designated P5-P11 by poly-(L-lysine)-agarose and aminohexyl Sepharose 4B chromatographies. These seven phosphatase activities were active toward nonprotein phosphoester. P7-P11 and to some extent P5 could also dephosphorylate a phosphoprotein. They displayed different enzyme kinetics. On the basis of activity peak, the apparent molecular mass as estimated by Sephadex G-200 column chromatography for P5 was 49 kDa; P6, 32 kDa; P7, 150 kDa; P8, 250 kDa; P9, 165 kDa; P10, 90 kDa and P11, 165 kDa. Immunoblot analysis indicated that P8-P11 may belong to PP2B family, whereas P7 may associate with PP2A. The phosphatases P7-P11 were found to be effective in the dephosphorylation of Alzheimer's disease abnormally hyperphosphorylated tau. The resulting dephosphorylated tau regained its activity in promoting the microtubule assembly, suggesting that P7-P11 might regulate the phosphorylation of tau protein in the brain.

  15. Zinc ions and alkaline pH alter the phosphorylation state of human erythrocyte membrane proteins

    Fennell, R.L. Jr.


    Since the phosphorylation state of the red cell membrane proteins in vitro is likely to be regulated by phosphorylation and dephosphorylation, this research was carried out to investigate the possible role of membrane-bound phosphatase activities. These studies were conducted with red blood cell ghosts and IOVs from normal individuals and from an individual with hereditary spherocytosis. In vitro phosphorylation with ({gamma}-{sup 32}P) ATP was conducted in the presence and the absence of Zn{sup ++}, or erythrocyte ghosts and IOVs were pretreated for 30 minutes at 37{degree}C and pH 7-11 in the presence and the absence of calf intestine alkaline phosphatase. The resulting phosphoproteins were analyzed by SDS-polyacrylamide gel electrophoresis, stained with Coomassie blue, and fluorographed. In the presence of Zn{sup ++}, the red blood ghosts, with or without pretreatment, demonstrated enhanced phosphorylation of membrane proteins, including band 4.2. Preincubation at pH 10 in the presence of absence of exogenous phosphatase further stimulates phosphorylation of these proteins. Under similar conditions, the erythrocyte membranes also demonstrated the ability to hydrolyze p-nitrophenyl phosphate and to remove {sup 32}P from red blood cell phosphoproteins.

  16. Effects icariin of on the alkline phosphatase activity of human periodontal ligament cells inhibited by LPS%淫羊藿苷对内毒素抑制牙周膜细胞ALP表达的影响

    姜瑛; 王祥; 许华; 刘英群


    Objective: To investigate the effects of icariin (ICA) on proliferation of huaman periodontal ligament cells (human periodontal ligament cells, hPDLC) and the alkaline phosphatase (alkaline phosphatase, ALP) activity inhibited by LPS. Method: HPDLC were cultured in virto and stimulated with icariin with different concentrations (0, 10-5、 10-6、 10-7、 10-8、 10-9 mol / L) for 96 hours.The ability of proliferation of HPDLC was detected by MTT method.The activity of alkaline phosphatase and the expression of alkaline phosphatase mRNA was seperately determined by p-Nitrophnyl phosphate (pNPP) method and RT-PCR after being treated with icariin at the concentration of 10-6 mol / L inhibited by LPS. Result: icariin had a dose-dependent effect on the proliferation the hPDLC in a suitable concentration range from l0-6-10-8 mol / L. The alkaline phosphatase activity was remarkably inhibited by incubation of PDLC with 10 μg / mL LPS and was improved in the presence of icariin at the concentration of 10-6 mol / L. Conclusion: Icariin have the effect on the proliferation of PDLC and improve the ALP activity inhibited by LPS, which could help for Peri—apical tissue regeneration.%目的:探讨淫羊藿苷对人牙周膜细胞(periodontal ligament cells,PDLC)增殖及对内毒素(Lipopolysaccharide,LPS)干扰下碱性磷酸酶(alkaline phosphatase,ALP)的影响.方法:原代培养PDLC,MTT法检测不同浓度(0、10-5~10-9 mol/L)淫羊藿苷对PDLC增殖的影响;RT-PCR、对硝基苯酚法测定淫羊藿苷对LPS抑制PDLC的ALP mRNA表达和分泌的影响.结果:淫羊藿苷在一定浓度下(10-6~10-7 mol/L)可促进PDLC增殖;10 μg/mL LPS可抑制PDLC的ALP活性;加入10-6 mol/L淫羊藿苷干扰后,对LPS抑制PDLC的ALP有拮抗作用,可提高其mRNA表达和活性.结论:淫羊藿苷在一定浓度时可促进PDLC增殖;可能通过拮抗LPS抑制其ALP的活性.

  17. Handbook of Indigenous Foods Involving Alkaline Fermentation

    Sarkar, P.K.; Nout, M.J.R.


    This book details the basic approaches of alkaline fermentation, provides a brief history, and offers an overview of the subject. The book discusses the diversity of indigenous fermented foods involving an alkaline reaction, as well as the taxonomy, ecology, physiology, and genetics of predominant m

  18. Structural mechanisms of plant glucan phosphatases in starch metabolism.

    Meekins, David A; Vander Kooi, Craig W; Gentry, Matthew S


    Glucan phosphatases are a recently discovered class of enzymes that dephosphorylate starch and glycogen, thereby regulating energy metabolism. Plant genomes encode two glucan phosphatases, called Starch EXcess4 (SEX4) and Like Sex Four2 (LSF2), that regulate starch metabolism by selectively dephosphorylating glucose moieties within starch glucan chains. Recently, the structures of both SEX4 and LSF2 were determined, with and without phosphoglucan products bound, revealing the mechanism for their unique activities. This review explores the structural and enzymatic features of the plant glucan phosphatases, and outlines how they are uniquely adapted to perform their cellular functions. We outline the physical mechanisms used by SEX4 and LSF2 to interact with starch glucans: SEX4 binds glucan chains via a continuous glucan-binding platform comprising its dual-specificity phosphatase domain and carbohydrate-binding module, while LSF2 utilizes surface binding sites. SEX4 and LSF2 both contain a unique network of aromatic residues in their catalytic dual-specificity phosphatase domains that serve as glucan engagement platforms and are unique to the glucan phosphatases. We also discuss the phosphoglucan substrate specificities inherent to SEX4 and LSF2, and outline structural features within the active site that govern glucan orientation. This review defines the structural mechanism of the plant glucan phosphatases with respect to phosphatases, starch metabolism and protein-glucan interaction, thereby providing a framework for their application in both agricultural and industrial settings.

  19. P depletion and activity of phosphatases in the rhizosphere of mycorrhizal and non-mycorrhizal cucumber (Cucumis Sativus L.)

    Joner, E.J.; Magid, J.; Gahoonia, T.S.;


    was sectioned in a freezing microtome and analyzed for extracellular acid (pH 5.2) and alkaline (pH 8.5) phosphatase activity as well as depletion of NaHCO-3-extractable inorganic P (P-i) and P-o. Roots and mycorrhizal hyphae depleted the soil of P-i but did not influence the concentration of P-o in spite......An experiment was set up to test the ability of arbuscular mycorrhizal (AM) roots and hyphae to produce extracellular phosphatases and to study the relationship between phosphatase activity and soil organic P (P-o). Non-mycorrhizal cucumber and cucumber in symbiosis with either of two mycorrhizal...... fungi were grown in a sandy loam-sand mixture in three-compartment pots. Plant roots were separated from two consecutively adjoining compartments, first by a 37 m mesh excluding roots and subsequently by a 0.45 m membrane excluding mycorrhizal hyphae. Soil from the two root-free compartments...


    Beata Kuziemska


    Full Text Available A study was carried out on soil following a two-year pot experiment that was conducted in 2009–2010, in three repetitions in Siedlce. The experiment included the following factors: 1 – amount of Ni in soil (0, 75, 150 and 225 mg·kg-1 soil by applying an aqueous NiSO4·7H2O solution; 2 – liming (0 and Ca according to 1 Hh as CaCO3; 3 – organic waste products (rye straw at a dose of 4 t·ha-1 and brown coal at a dose of 40 t·ha-1. In each experimental year, orchard grass was the test plant and four swaths were harvested. The activities of acidic and alkaline phosphatase, pH and the content of carbon in organic compounds were determined in the soil samples collected after each grass swath and in each experimental year. It was found that Ni at 75 mg·kg-1 soil activated the enzymes under study, whereas higher doses caused their statistically-confirmed inactivation. The lowest activity of the investigated enzymes was detected in soil supplemented with 225 Ni·kg-1 soil. Liming caused an increase in the activity of alkaline phosphatase and a reduction in the activity of acidic phosphatase. Straw and brown coal induced a substantial increase in the activity of both enzymes in the tested soil samples. Both liming and straw and carbon eliminated the negative effect of higher nickel doses on the activity of the enzymes under study.

  1. Inositol monophosphate phosphatase genes of Mycobacterium tuberculosis

    Parish Tanya


    Full Text Available Abstract Background Mycobacteria use inositol in phosphatidylinositol, for anchoring lipoarabinomannan (LAM, lipomannan (LM and phosphatidylinosotol mannosides (PIMs in the cell envelope, and for the production of mycothiol, which maintains the redox balance of the cell. Inositol is synthesized by conversion of glucose-6-phosphate to inositol-1-phosphate, followed by dephosphorylation by inositol monophosphate phosphatases (IMPases to form myo-inositol. To gain insight into how Mycobacterium tuberculosis synthesises inositol we carried out genetic analysis of the four IMPase homologues that are present in the Mycobacterium tuberculosis genome. Results Mutants lacking either impA (Rv1604 or suhB (Rv2701c were isolated in the absence of exogenous inositol, and no differences in levels of PIMs, LM, LAM or mycothiol were observed. Mutagenesis of cysQ (Rv2131c was initially unsuccessful, but was possible when a porin-like gene of Mycobacterium smegmatis was expressed, and also by gene switching in the merodiploid strain. In contrast, we could only obtain mutations in impC (Rv3137 when a second functional copy was provided in trans, even when exogenous inositol was provided. Experiments to obtain a mutant in the presence of a second copy of impC containing an active-site mutation, in the presence of porin-like gene of M. smegmatis, or in the absence of inositol 1-phosphate synthase activity, were also unsuccessful. We showed that all four genes are expressed, although at different levels, and levels of inositol phosphatase activity did not fall significantly in any of the mutants obtained. Conclusions We have shown that neither impA, suhB nor cysQ is solely responsible for inositol synthesis. In contrast, we show that impC is essential for mycobacterial growth under the conditions we used, and suggest it may be required in the early stages of mycothiol synthesis.

  2. Comparison of enzyme-linked immunosorbent assay and radioimmunoassay for prostate-specific acid phosphatase in prostatic disease

    Griffiths, J.; Rippe, D.F.; Panfili, P.R.


    Results of an enzyme-linked immunosorbent assay (ELISA) are compared with those of a standard radioimmunoassay (RIA) for detection and quantitation of prostate-specific acid phosphatase (EC in serum. Control subjects, patients with benign prostatic hyperplasia, and patients in all four clinical stages of prostatic adenocarcinoma were tested. The upper limit of normal (95%of the population) by the ELISA was 2.0, and by the RIA was 2.2 In prostatic a denocarcinoma stage I (not detectable by digital rectal examination), ELISA was slightly more sensitive than RIA, but sensitivity was still relatively low (20%). As tumor mass increased (stages II through IV), the frequency of increased concentrations of prostatic acid phosphatase in serum also increased. We confirmed this increase in circulating enzyme in some cases of benign prostatic hyperplasia and suggest that this finding is related to either acinar cytolysis or an increase in acini size and number. Although prostate-specific acid phosphatase is not a cancer-specific enzyme, we conclude that its measurement may be of considerable value in monitoring prostatic disease.

  3. Voltage sensitive phosphatases: emerging kinship to protein tyrosine phosphatases from structure-function research

    Kirstin eHobiger


    Full Text Available The transmembrane protein Ci-VSP from the ascidian Ciona intestinalis was described as first member of a fascinating family of enzymes, the voltage sensitive phosphatases (VSPs. Ci-VSP and its voltage-activated homologs from other species are stimulated by positive membrane potentials and dephosphorylate the head groups of negatively charged phosphoinositide phosphates (PIPs. In doing so, VSPs act as control centers at the cytosolic membrane surface, because they intervene in signaling cascades that are mediated by PIP lipids. The characteristic motif CX5RT/S in the active site classifies VSPs as members of the huge family of cysteine-based protein tyrosine phosphatases (PTPs. Although PTPs have already been well characterized regarding both, structure and function, their relationship to VSPs has drawn only limited attention so far. Therefore, the intention of this review is to give a short overview about the extensive knowledge about PTPs in relation to the facts known about VSPs. Here, we concentrate on the structural features of the catalytic domain which are similar between both classes of phosphatases and their consequences for the enzymatic function. By discussing results obtained from crystal structures, molecular dynamics simulations, and mutagenesis studies, a possible mechanism for the catalytic cycle of VSPs is presented based on that one proposed for PTPs. In this way, we want to link the knowledge about the catalytic activity of VSPs and PTPs.

  4. Different designs of kinase-phosphatase interactions and phosphatase sequestration shapes the robustness and signal flow in the MAPK cascade

    Sarma Uddipan


    Full Text Available Abstract Background The three layer mitogen activated protein kinase (MAPK signaling cascade exhibits different designs of interactions between its kinases and phosphatases. While the sequential interactions between the three kinases of the cascade are tightly preserved, the phosphatases of the cascade, such as MKP3 and PP2A, exhibit relatively diverse interactions with their substrate kinases. Additionally, the kinases of the MAPK cascade can also sequester their phosphatases. Thus, each topologically distinct interaction design of kinases and phosphatases could exhibit unique signal processing characteristics, and the presence of phosphatase sequestration may lead to further fine tuning of the propagated signal. Results We have built four architecturally distinct types of models of the MAPK cascade, each model with identical kinase-kinase interactions but unique kinases-phosphatases interactions. Our simulations unravelled that MAPK cascade’s robustness to external perturbations is a function of nature of interaction between its kinases and phosphatases. The cascade’s output robustness was enhanced when phosphatases were sequestrated by their target kinases. We uncovered a novel implicit/hidden negative feedback loop from the phosphatase MKP3 to its upstream kinase Raf-1, in a cascade resembling the B cell MAPK cascade. Notably, strength of the feedback loop was reciprocal to the strength of phosphatases’ sequestration and stronger sequestration abolished the feedback loop completely. An experimental method to verify the presence of the feedback loop is also proposed. We further showed, when the models were activated by transient signal, memory (total time taken by the cascade output to reach its unstimulated level after removal of signal of a cascade was determined by the specific designs of interaction among its kinases and phosphatases. Conclusions Differences in interaction designs among the kinases and phosphatases can

  5. Alkaline solution neutralization capacity of soil.

    Asakura, Hiroshi; Sakanakura, Hirofumi; Matsuto, Toshihiko


    Alkaline eluate from municipal solid waste (MSW) incineration residue deposited in landfill alkalizes waste and soil layers. From the viewpoint of accelerating stability and preventing heavy metal elution, pH of the landfill layer (waste and daily cover soil) should be controlled. On the other hand, pH of leachate from existing MSW landfill sites is usually approximately neutral. One of the reasons is that daily cover soil can neutralize alkaline solution containing Ca(2+) as cation. However, in landfill layer where various types of wastes and reactions should be taken into consideration, the ability to neutralize alkaline solutions other than Ca(OH)(2) by soil should be evaluated. In this study, the neutralization capacities of various types of soils were measured using Ca(OH)(2) and NaOH solutions. Each soil used in this study showed approximately the same capacity to neutralize both alkaline solutions of Ca(OH)(2) and NaOH. The cation exchange capacity was less than 30% of the maximum alkali neutralization capacity obtained by the titration test. The mechanism of neutralization by the pH-dependent charge can explain the same neutralization capacities of the soils. Although further investigation on the neutralization capacity of the soils for alkaline substances other than NaOH is required, daily cover soil could serve as a buffer zone for alkaline leachates containing Ca(OH)(2) or other alkaline substances.

  6. Relationship between serum leptin levels and bone mineral density and bone metabolic markers in patients on hemodialysis.

    Ahmadi, Farokhlagha; Salari, Sina; Maziar, Sima; Esfahanian, Fateme; Khazaeipour, Zahra; Ranjbarnovin, Neda


    Leptin is the protein product of the obesity gene, which is produced in fat tissue. It was originally thought to be involved only in the regulation of food intake and energy balance. We aimed to investigate the relationship of serum leptin levels with bone mineral density (BMD) and biochemical markers of bone turnover in patients on hemodialysis (HD). This study included 72 patients (43 males and 29 females), whose mean age was 55.1 ± 11.4 years, mean body mass index was 23.13 ± 2.75 kg/m 2 and mean duration on HD was 5 ± 3.4 years. The BMD values were calculated using dual-energy X-ray absorptiometry (DEXA) at the femoral neck and lumbar spine. Blood samples were taken for leptin, intact parathyroid hormone (I-PTH), bone alkaline phosphatase (BAP), calcium (Ca), phosphate (P) and albumin. The leptin levels were higher in females than in males (22.3 ± 19.6 vs 20.8 ± 23), but this difference was not significant. The serum leptin level had a strong positive correlation with Ca levels in the female patients (r = 0.659 and P = 0.01) and a negative correlation with albumin levels (r = -0.461 and P = 0.01). No correlation was found with age, BMI, duration on dialysis, BMD and serum levels of PTH, BAP and P for the entire patient group or either gender separately. The serum leptin level was significantly lower in females with PTH >300 pg/mL when compared with patients with PTH = 100-300 pg/mL (86 ± 85 vs 47 ± 48) (P = 0.011).Women with BAP leptin than those with BAP 300-600 IU/L (P = 0.024). Women with Ca leptin levels compared with those with Ca levels of 8.5-10.5 mg/dL (P = 0.011). There was no significant difference between the two genders among variables such as age, BMI, duration on dialysis, serum leptin, I-PTH, Ca, P, BAP, albumin and BMD of the femoral neck and lumbar spine.

  7. Trends of elevated parathormone serum titers in hemodialysis patients on intensive therapy for bone disease: A multicenter study

    Ayman Karkar


    Full Text Available To determine the prevalence of controlled parathyroid hormone (PTH serum levels with intensified therapy for chronic kidney disease mineral and bone disorder (CKD-MBD in the dialysis population, we studied 563 chronic hemodialysis patients recruited from three different dialysis centers from three different major cities in the Kingdom of Saudi Arabia. The trend of the routine monthly chemistries related to CKD-MBD was evaluated besides the whole-molecule PTH serum levels over 28 months (January 2011 to April 2013. The cost ratios of the medications to the estimated dialysis total cost were calculated. There were 323 (57.4% males in the study, and the mean age of the patients was 50.2 ± 15.2 years; 371 (65.9% patients were initiated on dialysis before 2011. The causes of the original kidney disease included diabetes mellitus in 163 (29% patients. Parathyroidectomy was performed in 23 (4.1% patients and only six (23% patients underwent the operation during the study period; most of the parathyroidectomies (69% were performed before 2011. The trend of the medians of monthly serum levels of calcium, phosphorus, albumin, bicarbonate, alkaline phosphatase, serum levels of PTH and vitamin D25 assays showed better control of the levels with time. The added cost of cinacalcet was more significant than the other drugs, including vitamin D and phosphate binders, but the cost was minimal in comparison with the whole dialysis bill. The ratios of the discontinuation rates to the total patient-months of treatment for the different drugs were in the range of 3-4% and mostly due to transient overdosing of medications. We conclude that the trends of the median serum levels of PTH and related minerals in the CKD patients in our dialysis patients suggested a good inclination toward control and prevention of the vascular calcifications prevalent in the CKD-MBD. The popularity of use of new drugs such as cinacalcet is promising and does not seem to add much to the

  8. Detection of serum AFB1-lysine adduct in Malaysia and its association with liver and kidney functions.

    Mohd Redzwan, S; Rosita, Jamaluddin; Mohd Sokhini, A M; Nurul 'Aqilah, A R; Wang, Jia-Sheng; Kang, Min-Su; Zuraini, Ahmad


    Aflatoxin is ubiquitously found in many foodstuffs and produced by Aspergillus species of fungi. Of many aflatoxin metabolites, AFB1 is classified by the International Agency for Research on Cancer (IARC) as group one carcinogen and linked to the development of hepatocellular carcinoma (HCC). The study on molecular biomarker of aflatoxin provides a better assessment on the extent of human exposure to aflatoxin. In Malaysia, the occurrences of aflatoxin-contaminated foods have been documented, but there is a lack of data on human exposure to aflatoxin. Hence, this study investigated the occurrence of AFB1-lysine adduct in serum samples and its association with liver and kidney functions. 5ml fasting blood samples were collected from seventy-one subjects (n=71) for the measurement of AFB1-lysine adduct, albumin, total bilirubin, AST (aspartate aminotransferase), ALT (alanine transaminase), ALP (alkaline phosphatase), GGT (gamma-glutamyl transpeptidase), creatinine and BUN (blood urea nitrogen). The AFB1-lysine adduct was detected in all serum samples (100% detection rate) with a mean of 6.85±3.20pg/mg albumin (range: 1.13-18.85pg/mg albumin). Male subjects (mean: 8.03±3.41pg/mg albumin) had significantly higher adduct levels than female subjects (mean: 5.64±2.46pg/mg albumin) (padduct levels greater than average (>6.85pg/mg albumin) had significantly elevated level of total bilirubin (padduct. This study provides a valuable insight on human exposure to aflatoxin in Malaysia. Given that aflatoxin can pose serious problem to the health, intervention strategies should be implemented to limit/reduce human exposure to aflatoxin. Besides, a study with a big sample size should be warranted in order to assess aflatoxin exposure in the general population of Malaysia.

  9. Effects of dietary yeastβ-glucan on nutrient digestibility and serum proifles in pre-ruminant Holstein calves

    MA Tao; TU Yan; ZHANG Nai-feng; GUO Jiang-peng; DENG Kai-dong; ZHOU Yi; YUN Qiang; DIAO Qi-yu


    This study aimed to investigate the effects of dietary supplementation of yeastβ-glucan on the nutrient digestibility and serum proifles in pre-ruminant Holstein calves. Forty-two neonatal Holstein calves ((39.6±4.2) kg) were randomly al otted to six groups, and each was offered one of the fol owing diets:a basal diet (control) or the basal diet supplemented with 25, 50, 75, 100 or 200 mg of yeastβ-glucan kg–1 feed (dry matter basis). The basal diet consisted of a milk replacer and a starter feed. The trial lasted for 56 d. Two digestibility trials were conducted from d 14 to 20 and from d 42 to 48. Blood samples were col ected on d 0, 14, 28 and 42 for serum proifle analyses. On d 56, three calves from each group were slaughtered, and intestinal samples were col ected to assess the vil ous height, crypt depth and mucosal thickness. Although feed intake was not affected by dietary treatment (P>0.05), the average daily gain (ADG) and gain-to-feed ratios were higher (P0.05). Compared with the control group, supplementation of yeastβ-glucan decreased (P0.05). The supplementation of yeastβ-glucan stimu-lated the enzymatic activity of alkaline phosphatase (ALP) (P<0.05) compared with the control group. The lysozyme (LYZ) concentration increased quadratical y (P<0.05) with increasing yeastβ-glucan levels. The results suggested that dietary supplementation of yeastβ-glucan at 75 mg kg–1 feed improved nutrient digestibility, enhanced immunity by increasing the immunoglobulin concentration and stimulating ALP, and exerted no adverse effects on metabolism in pre-ruminant calves.

  10. Relationship between Body Mass Composition, Bone Mineral Density, Skin Fibrosis and 25(OH Vitamin D Serum Levels in Systemic Sclerosis.

    Addolorata Corrado

    Full Text Available A reduced bone mineral density (BMD is observed in several rheumatic autoimmune diseases, including Systemic Sclerosis (SSc; nevertheless, data concerning the possible determinants of bone loss in this disease are not fully investigated. The aim of this study is to evaluate the relationship between BMD, body mass composition, skin sclerosis and serum Vitamin D levels in two subsets of SSc patients. 64 post-menopausal SSc patients, classified as limited cutaneous (lcSSc or diffuse cutaneous (dcSSc SSc, were studied. As control, 35 healthy post-menopausal women were recruited. Clinical parameters were evaluated, including the extent of skin involvement. BMD at lumbar spine, hip, femoral neck and body mass composition were determined by dual-energy X-ray absorptiometry. Serum calcium, phosphorus, alkaline phosphatase, urine pyridinium cross-links, intact parathyroid hormone and 25-hydroxyvitamin D (25OHD were measured. BMD at spine, femoral neck and total hip was significantly lower in SSc patients compared to controls. In dcSSc subset, BMD at spine, femoral neck and total hip was significantly lower compared to lcSSc. No differences in both fat and lean mass were found in the three study groups even if patients with dcSSc showed a slightly lower total body mass compared to healthy controls. Total mineral content was significantly reduced in dSSc compared to both healthy subjects and lcSSc group. Hypovitaminosis D was observed both in healthy post-menopausal women and in SSc patients, but 25OHD levels were significantly lower in dcSSc compared to lcSSc and inversely correlated with the extent of skin thickness. These results support the hypothesis that the extent of skin involvement in SSc patients could be an important factor in determining low circulating levels of 25OHD, which in turn could play a significant role in the reduction of BMD and total mineral content.

  11. Effect of non-phytate phosphorus levels and phytase sources on the growth performance, serum biochemical and tibial parameters of broiler chickens

    Xian R. Jiang


    Full Text Available A 3×3 fattorial arrangement with dietary non-phytate phosphorus (NPP levels and phytase sources (3- and 6-phytase was conducted to evaluate the effects of NPP levels, phytase sources and their possible interactions on growth performance, serum biochemical and tibia parameters of broiler chickens from hatch to 42 days of age. A total of 540 1-day-old Arbor Acres male broiler chicks were randomly allocated into nine dietary treatments, each containing 5 replicates pens with 12 birds per pen. Interaction was statistically significant in the performance till day 21 of trial, supplementation of low NPP diet decreased body weight (BW (P<0.001, depressed average daily gain (ADG (P<0.001 and deteriorated average daily feed intake (ADFI (P<0.001 over day 42. During the 8-to-21-day period, even if interaction between NPP levels and phytase sources was significant (P<0.01, BW, ADG and ADFI always increased due to dietary supplementation of phytase, with source not differing. Dietary high NPP enhanced serum calcium and P concentrations on day 21 and 42 (linear contrast, P<0.01, while decreased alkaline phosphatase (AKP activity on day 42 (linear contrast, P<0.001, and interaction was not significant. Both dietary sources of phytase decreased serum AKP activities on day 42 (P<0.001, and urea nitrogen content on day 21 (P<0.01 and 42 (P<0.001. Both phytase improved ash percentage on day 21 and P content in tibia at 21 and 42 days of age (P<0.001. The results confirmed that dietary supplementation of phytase may enhance P availability during the 8-to-21-day period. Nevertheless, no difference between the two phytase sources was observed.

  12. The Effects of Varying Concentrations of Dietary Protein and Fat on Blood Gas, Hematologic Serum Chemistry, and Body Temperature Before and After Exercise in Labrador Retrievers.

    Ober, John; Gillette, Robert L; Angle, Thomas Craig; Haney, Pamela; Fletcher, Daniel J; Wakshlag, Joseph J


    Optimal dietary protocols for the athletic canine are often defined by requirements for endurance athletes that do not always translate into optimal dietary interventions for all canine athletes. Prior research studying detection dogs suggests that dietary fat sources can influence olfaction; however, as fat is added to the diet the protein calories can be diminished potentially resulting in decreased red blood cell counts or albumin status. Optimal macronutrient profile for detection dogs may be different considering the unique work they engage in. To study a calorically low protein: high fat (18:57% ME), high protein: high fat (27:57% ME), and high protein: low fat (27:32% ME) approach to feeding, 17 dogs were provided various diets in a 3 × 3 cross over design. Dogs were exercised on a treadmill and blood was taken pre-exercise, immediately post-exercise, 10- and 20-min post-exercise to assess complete blood count, serum chemistry, blood gases, and cortisol; as well as rectal and core body temperature. Exercise induced a decrease in serum phosphorus, potassium, and increases in non-esterified fatty acids and cortisol typical of moderate exercise bouts. A complete and balanced high protein: high-fat diet (27:57% ME) induced decreases in serum cortisol and alkaline phosphatase. Corn oil top dressed low protein: high-fat diet (18:57% ME) induced a slightly better thermal recovery than a complete and balanced high protein: high fat diet and a high protein: low fat (27%:32% ME) diet suggesting some mild advantages when using the low protein: high fat diet that warrant further investigation regarding optimal protein and fat calories and thermal recovery.

  13. Structure-Function Analysis of the 3' Phosphatase Component of T4 Polynucleotide Kinase/phosphatase

    Zhu,H.; Smith, P.; Wang, L.; Shuman, S.


    T4 polynucleotide kinase/phosphatase (Pnkp) exemplifies a family of bifunctional enzymes with 5'-kinase and 3' phosphatase activities that function in nucleic acid repair. T4 Pnkp is a homotetramer of a 301-aa polypeptide, which consists of an N-terminal kinase domain of the P-loop phosphotransferase superfamily and a C-terminal phosphatase domain of the DxD acylphosphatase superfamily. The homotetramer is formed via pairs of phosphatase-phosphatase and kinase-kinase homodimer interfaces. Here we identify four side chains-Asp187, Ser211, Lys258, and Asp277-that are required for 3' phosphatase activity. Alanine mutations at these positions abolished phosphatase activity without affecting kinase function or tetramerization. Conservative substitutions of asparagine or glutamate for Asp187 did not revive the 3' phosphatase, nor did arginine or glutamine substitutions for Lys258. Threonine in lieu of Ser211 and glutamate in lieu of Asp277 restored full activity, whereas asparagine at position 277 had no salutary effect. We report a 3.0 A crystal structure of the Pnkp tetramer, in which a sulfate ion is coordinated between Arg246 and Arg279 in a position that we propose mimics one of the penultimate phosphodiesters (5'NpNpNp-3') of the polynucleotide 3'-PO(4) substrate. The amalgam of mutational and structural data engenders a plausible catalytic mechanism for the phosphatase that includes covalent catalysis (via Asp165), general acid-base catalysis (via Asp167), metal coordination (by Asp165, Asp277 and Asp278), and transition state stabilization (via Lys258, Ser211, backbone amides, and the divalent cation). Other critical side chains play architectural roles (Arg176, Asp187, Arg213, Asp254). To probe the role of oligomerization in phosphatase function, we introduced six double-alanine cluster mutations at the phosphatase-phosphatase domain interface, two of which (R297A-Q295A and E292A-D300A) converted Pnkp from a tetramer to a dimer

  14. The SHP-2 tyrosine phosphatase: Signaling mechanisms and biological functions


    Cellular biological activities are tightly controlled by intracellular signaling processes initiated by extracellular signals.Protein tyrosine phosphatases, which remove phosphate groups from phosphorylated signaling molecules, play equally important tyrosine roles as protein tyrosine kinases in signal transduction.SHP-2, a cytoplasmic SH2 domain containing protein tyrosine phosphatase, is involved in the signaling pathways of a variety of growth factors and cytokines. Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus, and is a critical intracellular regulator in mediating cell proliferation and differentiation.

  15. Short-term effect of vermicompost application on biological properties of an alkaline soil with high lime content from Mediterranean region of Turkey.

    Uz, Ilker; Tavali, Ismail Emrah


    This study was conducted to investigate direct short-term impact of vermicompost on some soil biological properties by monitoring changes after addition of vermicompost as compared to farmyard manure in an alkaline soil with high lime content from semiarid Mediterranean region of Turkey. For this purpose, mixtures of soil and organic fertilizers in different doses were incubated under greenhouse condition. Soil samples collected in regular intervals were analyzed for biological parameters including dehydrogenase, β-glucosidase, urease, alkaline phosphatase activities, and total number of aerobic mesophilic bacteria. Even though soil dehydrogenase activity appeared to be dose-independent based on overall evaluation, organic amendments were found to elevate dehydrogenase activity when sampling periods are evaluated individually. β-glucosidase, urease, alkaline phosphatase activity, and aerobic mesophilic bacterial numbers in vermicompost treatments fluctuated but remained significantly above the control. A slight but statistically significant difference was detected between organic amendments in terms of urease activity. Vermicompost appeared to more significantly increase bacterial number in soil. Clearly, vermicompost has a potential to be used as an alternative to farmyard manure to improve and maintain soil biological activity in alkaline calcareous soils from the Mediterranean region of Turkey. Further studies are needed to assess its full potential for these soils.

  16. Protein tyrosine phosphatases: structure-function relationships.

    Tabernero, Lydia; Aricescu, A Radu; Jones, E Yvonne; Szedlacsek, Stefan E


    Structural analysis of protein tyrosine phosphatases (PTPs) has expanded considerably in the last several years, producing more than 200 structures in this class of enzymes (from 35 different proteins and their complexes with ligands). The small-medium size of the catalytic domain of approximately 280 residues plus a very compact fold makes it amenable to cloning and overexpression in bacterial systems thus facilitating crystallographic analysis. The low molecular weight PTPs being even smaller, approximately 150 residues, are also perfect targets for NMR analysis. The availability of different structures and complexes of PTPs with substrates and inhibitors has provided a wealth of information with profound effects in the way we understand their biological functions. Developments in mammalian expression technology recently led to the first crystal structure of a receptor-like PTP extracellular region. Altogether, the PTP structural work significantly advanced our knowledge regarding the architecture, regulation and substrate specificity of these enzymes. In this review, we compile the most prominent structural traits that characterize PTPs and their complexes with ligands. We discuss how the data can be used to design further functional experiments and as a basis for drug design given that many PTPs are now considered strategic therapeutic targets for human diseases such as diabetes and cancer.

  17. The RCN1-encoded A subunit of protein phosphatase 2A increases phosphatase activity in vivo

    Deruere, J.; Jackson, K.; Garbers, C.; Soll, D.; Delong, A.; Evans, M. L. (Principal Investigator)


    Protein phosphatase 2A (PP2A), a heterotrimeric serine/threonine-specific protein phosphatase, comprises a catalytic C subunit and two distinct regulatory subunits, A and B. The RCN1 gene encodes one of three A regulatory subunits in Arabidopsis thaliana. A T-DNA insertion mutation at this locus impairs root curling, seedling organ elongation and apical hypocotyl hook formation. We have used in vivo and in vitro assays to gauge the impact of the rcn1 mutation on PP2A activity in seedlings. PP2A activity is decreased in extracts from rcn1 mutant seedlings, and this decrease is not due to a reduction in catalytic subunit expression. Roots of mutant seedlings exhibit increased sensitivity to the phosphatase inhibitors okadaic acid and cantharidin in organ elongation assays. Shoots of dark-grown, but not light-grown seedlings also show increased inhibitor sensitivity. Furthermore, cantharidin treatment of wild-type seedlings mimics the rcn1 defect in root curling, root waving and hypocotyl hook formation assays. In roots of wild-type seedlings, RCN1 mRNA is expressed at high levels in root tips, and accumulates to lower levels in the pericycle and lateral root primordia. In shoots, RCN1 is expressed in the apical hook and the basal, rapidly elongating cells in etiolated hypocotyls, and in the shoot meristem and leaf primordia of light-grown seedlings. Our results show that the wild-type RCN1-encoded A subunit functions as a positive regulator of the PP2A holoenzyme, increasing activity towards substrates involved in organ elongation and differential cell elongation responses such as root curling.

  18. Basic data on the hematology, serum biochemistry, urology, and organ weights of beagle dogs.

    Choi, So-Young; Hwang, Jae-Sik; Kim, Ill-Hwa; Hwang, Dae-Yeon; Kang, Hyun-Gu


    This study was conducted to provide basic data on physiological and hematological characteristics, and organ weights of beagle dogs. A total of 237 beagle dogs were used to determine differences in physiological and hematological parameters, and organ weights depending on sex and age. The respiratory rate of both sexes tended to increase as they grew older and the female heart rate was slightly higher than that of males. Male and female body weights increased rapidly to 33 weeks old followed by a gradual increase to 41-weeks-old. The relative weight of the brain was negatively correlated with body weight, whereas the weight of reproductive organs was positively correlated with body weight. The platelet count of female dogs was slightly higher than that of males. The red blood cell, hemoglobin, and hematocrit of both sexes increased non-significantly with age. In the leukocyte differential count, the neutrophils, and eosinophils of both sexes tended to increase as they grew older, whereas basophils, lymphocytes, and monocytes decreased. In the serum biochemical profiles, alkaline phosphatase was slightly higher in males than females, while the total cholesterol of female dogs at 9-months-old was higher than that of males at the same age. Other biochemical components, including alanine aminotransferase, blood urea nitrogen, creatinine, triglyceride, and total protein increased non-significantly with age in both sexes. To conclude, we observe no significant physiological or hematological differences with sex or age, although decreasing and increasing trends were detected with some parameters. These data provide valuable reference indices of the normal physiological and hematological characteristics of beagle dogs, which should prove useful in toxicological and pharmacological studies.

  19. Chromogranin A as Serum Marker for Gastroenteropancreatic Neuroendocrine Tumors: A Single Center Experience and Literature Review

    Christoph J. Auernhammer


    Full Text Available The aim of this study was to assess the clinical sensitivities of the tumor markers chromogranin A (CgA, urinary 5-hydroxyindoleacetic acid (5-HIAA and alkaline phosphatase (AP in neuroendocrine tumors (NETs of the GastroEnteroPancreatic-(GEP- system depending on tumor primary location and metastatic spread. In a retrospective single-center ser