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Alkaline phosphatase isoenzymes in feline serum using an agarose gel alkaline phosphatase kit method.  

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Total serum alkaline phosphatase (ALP) activity is the product of the combined activity of isoenzymes from a number of tissue sources. In this study, a commercially available kit for electrophoretic separation of ALP isoenzymes in an agarose gel was used to separate ALP isoenzymes in feline tissue extracts and serum. Five separate bands of ALP activity were identified. These bands were numbered 1 to 5 with band 1 having the most anodal migration. The tissue of origin corresponding to the migr...

Horney, B. S.; Farmer, A. J.; Mackensie, A.; Honor, D. J.; Buczkowski, S.

1992-01-01

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Probable levetiracetam-related serum alkaline phosphatase elevation  

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Full Text Available Abstract Background Levetiracetam (LEV is an antiepileptic drug with a favorable tolerability and safety profile with little or no effect on liver function. Case presentation Here, we reported an epileptic pediatric patient who developed a significant elevation in serum alkaline phosphatase level (ALP during LEV monotherapy. Moreover, the serum ALP level was surprisingly decreased to normal after LEV discontinuation. The Naranjo Adverse Drug Reaction Probability Scale score was 6, indicating firstly LEV was a probable cause for the increased serum ALP. Conclusions Cautious usage and concerns of the LEV-associated potential ALP elevation should be considered when levetiracetam is prescribed to epilepsy patients, especially pediatric patients.

Xiong Nian

2012-09-01

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Serum gamma-glutamyltransferase and alkaline phosphatase in rheumatoid arthritis.  

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Serum gamma-glutamyltransferase (GGT) and alkaline phosphatase (AP) were assayed in 98 consecutive patients with rheumatoid arthritis. Twenty-three patients had increased GGT activities and 45 an increased AP activity. Twelve patients showed an increase in both enzyme activities and AP isoenzyme studies were performed on seven of this group. In three subjects an increase in the bone isoenzyme was observed and in three others the increase in activity was attributed to the liver isoenzyme. The ...

1982-01-01

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Serum alkaline phosphatase assay with paired emitter detector diode.  

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A simple multicommutated flow system based on optoelectronic detector, three valves and peristaltic pump only has been developed for photometric determination of alkaline phosphatase activity in human serum. A miniaturized, compact flow-through detector dedicated to selective photometric detection of product formed in the course of the enzyme assay has been constructed using two paired light emitting diodes. The proposed analytical procedure based on kinetic methodology of enzyme activity detection and stopped-flow methodology of two-point measurements eliminates interferences caused by intense color of real samples and impurities present in commercial reagents. After optimization the system allows reproducible, mechanized analysis of human serum in relatively short time (8-9 samples per hour). Volume of serum required for single determination is 0.05mL only. The system validated with real clinical samples is useful for determination of enzyme activity in human serum at physiological and pathological levels. PMID:22817939

Strzelak, Kamil; Koncki, Robert; Tymecki, Lukasz

2012-07-15

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Fulminant hepatic failure with virtually undetectable serum alkaline phosphatase.  

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Wilson's disease presenting as fulminant hepatic failure is a rare presentation that carries a high morbidity and mortality. We report a young patient who developed fulminant hepatic failure as the initial manifestation of Wilson's disease. Virtually undetectable serum alkaline phosphatase provided the first clue to the diagnosis. Our patient underwent a successful liver transplantation which is the only effective treatment in patients with Wilsonian fulminant hepatic failure. In this report, we discuss laboratory clues to the diagnosis of this form of Wilson's disease. Clinicians should have a high suspicion of Wilson's disease as any delay in diagnosis can be catastrophic. PMID:24697127

Shaikh, Gulvahid; Shareef, Mohammed; Rahman, Sayed; Mustacchia, Paul; Rubinstein, Sofia; Walerstein, Steven

2014-01-01

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Serum Alkaline Phosphatase Levels and Mortality of Chronic Hemodialysis Patients  

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Full Text Available Objective: Alkaline phosphatase (ALP is considered a biomarker of high bone turnover in hemodialysis (HD patients with secondary hyperparathyroidism. This study was conducted to determine whether high serum ALP levels are associated with increased all-cause mortality of HD patients. Patients and Methods: This was a retrospective cohort study conducted at a single center. The subjects were 195 patients on chronic HD therapy who were followed up for a 5 years, and relationships between their baseline data and outcomes were assessed statistically. The serum ALP level was used as the predictor, and the primary end point was all-cause mortality. Results: Based on the median serum ALP of 236 IU/L, the subjects were divided into a low-ALP group (<236 IU/l and a high-ALP group (?236 IU/l. The high-ALP group was older and had a longer dialysis vintage, lower serum phosphorus concentrations, and higher serum parathyroid hormone levels, and they also had lower serum albumin levels and higher C-reactive protein values. In a multivariate Cox model in which the baseline serum ALP levels were used adjusted for age, gender, HD vintage, comorbidity, bone metabolism parameters, and serum liver enzyme levels, each doubling of the serum ALP level was associated with a significant increase in the hazard of all-cause mortality (hazard ratio 10.70, 95% CI 1.53 - 74.24. Conclusion: High baseline serum ALP levels are associated with increased mortality of HD patients, independent of bone metabolism parameters and serum liver enzyme levels. ALP is a potential target for the treatment of HD patients.

Tetsuri Yamashita

2011-09-01

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Wnt/?-Catenin Expression Does Not Correlate with Serum Alkaline Phosphatase Concentration in Canine Osteosarcoma Patients  

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Osteosarcoma is an aggressive malignancy of the bone and an increase in serum alkaline phosphatase concentration has clinical prognostic value in both humans and canines. Increased serum alkaline phosphatase concentration at the time of diagnosis has been associated with poorer outcomes for osteosarcoma patients. The biology underlying this negative prognostic factor is poorly understood. Given that activation of the Wnt signaling pathway has been associated with alkaline phosphatase expressi...

Piskun, Caroline M.; Muthuswamy, Anantharaman; Huelsmeyer, Michael K.; Thompson, Victoria; Stein, Timothy J.

2011-01-01

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Clinical significance of serum high-molecular-mass alkaline phosphatase, alkaline phosphatase-lipoprotein-X complex, and intestinal variant alkaline phosphatase.  

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This paper is a study to identify the clinical significance of high-molecular-mass alkaline phosphatase (ALP:E:C..3.1.3.1.), ALP-lipoprotein-X complex (LP-X) and intestinal variant ALP. We used cellulose acetate and agarose gels and techniques including wheat germ lectin, cetavlon-diethyl ether, thermostatability, neuraminidase and L-phenylalanine to improve the electrophoretic separation of the alkaline phosphatase isoenzymes. Patients' serum samples were electrophoresed from a diverse group of individuals ill with cholestasis, neoplastic disease metastatic to the liver, hepatocellular carcinoma, cirrhosis, diabetes mellitus, and chronic renal disease. Agarose gels provided better separation of ALP isoenzymes than cellulose acetate gels. The results also indicated that high-molecular mass ALP is present in patient's serum in conditions associated with cholestasis especially caused by hepatic malignancy. High-molecular mass ALP was frequently found to co-exist with the liver isoenzyme and LP-XALP complex. The intestinal variant was identified in patients with malignancy, cirrhosis, chronic renal disease and diabetes mellitus. Intestinal ALP coexisted concomitantly with a variant intestinal ALP. Intestinal variant ALP is most likely composed of intestinal ALP attached to a cellular membrane-binding domain, or may be an artifact produced by neuraminidase incubation. PMID:8046546

Wolf, P L

1994-01-01

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Correlation between MRI appearances and serum alkaline phosphatase levels in Osteogenic sarcoma  

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Introduction Magnetic resonance imaging is a pictorial depiction of the patho-anatomy of a bony lesion. As different parts of the tumor in Osteogenic sarcoma concurrently undergo various biological processes i.e. osteoblastic new bone formation, cell death, necrosis, bony destruction and revascularization etc., we hypothesized that the image seen in MRI could be used to assess the tumor behavior at that time. This study was done as a preliminary study with the aim to find whether the MRI pictures can have identifiable patterns and if present whether they can be linked to biological behavior. We could identify 2 distinct patterns in T2 weighted images which correlated well with serum alkaline phosphatase a serum marker and the duration of symptoms and so we are reporting our observations. Material and method T2 weighted MRI pictures of 15 cases of Osteogenic sarcoma were studied for identifiable patterns in matrix. These patterns if found were to be linked to biological behavior in the form of serum alkaline phosphatase levels and duration of symptoms. Results We could identify 2 unique patterns named by us as Group 1 Heterogeneous type (4 cases) which had a raised serum alkaline phosphatase level and had a history at presentation of less than 3 months duration. In Group 2 homogenous type (5 cases) the serum alkaline phosphatase levels were low and the cases presented after 6 months. As we could not identify any logical pattern in rest of the cases we labeled them as miscellaneous. Discussion MRI patterns can be used as markers of disease activity as there are 2 clear poles correlating well with serum alkaline phosphatase levels (high or low). Intermediate patterns may be the natural biological behavior and waxing and waning of the tumor disease activity.

Agrawal, Alok C.; Raza, H.K.T.; Sharma, Amit

2012-01-01

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Serum calcium and phosphorus concentration and alkaline phosphatase activity in healthy children during growth and development  

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Introduction. Many changes happen during growth and development in an organism as a result of important hormone changes, especially biohumoral ones. These changes make a problem when interpreting biochemical results in pediatric population. The most important changes are intensive calcium and phosphorus metabolic turnover in bone tissue with changes in alkaline phosphatase activity as a result of osteoblast activity. The aim of this study was to follow the serum calcium and phosphorus concent...

Savi? Ljiljana; Savi? Dejan

2008-01-01

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The effects of Zinc supplementation on serum zinc, alkaline phosphatase activity and fracture healing of bones  

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Objective was to determine the effect of zinc supplementation on callus information, serum zinc and alkaline phosphatase activity in humans. This randomized, double-blind, placebo controlled clinical trial was conducted on 60 patients with traumatic bone fracture referred to Shohada Hospital of Tabriz, Iran from August to December 2007. Subjects were randomly divided into 2 groups: cases (n=30), receiving one capsule of zinc sulfate consists of 50 mg zinc each day and the controls (n=30), receiving placebo for 60 days. Individual and clinical information was determined by a questionnaire: nutritional intake by 3 days food records at the beginning and the end of trial. Serum zinc and alkaline phosphatase was measured by atomic absorption spectroscopy and by enzymatic method. Callus information during fracture healing was evaluated by radiography of the bone. There was no significant difference in physical activity, gender, age, type of fractures and nutrient intake, between the 2 groups. The administration of zinc caused a significant elevation of serum zinc and alkaline phosphatase activity. Assessment of bone x-rays showed a significant progress in callus formation in cases compared to the controls. This study shows that zinc supplementation can stimulate fracture healing, however, it needs further study. (author)

2009-01-01

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Efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels in distinguishing exudates from transudates  

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Full Text Available The objective of present study was to evaluate the efficacy of pleural fluid alkaline phosphatase and its ratio to serum levels to classify pleural fluids. A total of 80 patients were divided in transudates and exudates on the basis of extensive clinical, radiological and biochemical evaluation. The efficacy of pleural fluid alkaline phosphatase (P ALP and pleural fluid / serum alkaline phosphatase ratio (P/S ALP assessment along with that of Light?s criteria to accurately classify transudates and exudates were analyzed. Up to 89% transudates misclassified by Light?s criteria were correctly classified by pleural fluid alkaline phosphatase (P ALP and pleural fluid/serum alkaline phosphatase ratio (P/S ALP evaluation. Similarly 92% exudates misclassified by Light?s criteria were correctly classified by pleural fluid alkaline phosphatase (P ALP and pleural fluid/serum alkaline phosphatase ratio (P/S ALP evaluation. By applying a cut off value of 40.0 IU for P ALP, a sensitivity of 85% and specificity of 75% was found. For P/S ALP, applying a cut off value of 0.25 a sensitivity of 85% and specificity of 80% was found. Both P ALP and P/S ALP had a PPV of 92%. However, their respective NPV were 63% and 70%.

Gupta K

2004-01-01

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Relation of serum alkaline phosphatase to liver scintigram in patients with hepatocellular carcinoma  

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Serum Alkaline Phosphatase (ALP) was studied in relation to liver scintigrams of 54 patients with hepatocellular carcinoma. The ALP activity was higher with larger tumors and in multiple tumors. Within the single tumor group, the activity was higher when the tumor was located in the hilum than in the periphery. The incidence of ALP-1 isoenzyme (bile ALP) roughly paralleled the total ALP activity. These results suggest that the variation of serum ALP seen in each individual patients with hepatocellular carcinoma reflects the volume of cholestatic liver tissue, which is changed by the number, size and localization of the tumor nodules in the liver. (orig.)

1982-01-01

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Relationship between serum heat-stable alkaline phosphatase level and pregnancy  

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Serum heat-stable alkaline phosphatase (HSAP) level in 649 cases of normal pregnancy and 164 cases of high-risk pregnancy is measured by radioimmunoassay (RIA). The results indicate that the HSAP level in normal pregnancy 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 fetal distress, the HSAP level in the mother's serum is also low. In 53 cases of intrahepatic cholestasis of pregnancy, the HSAP level is 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 pregnancy

1998-02-01

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Correlation Between Serum Level Parathormone, Alkaline Phosphatase, Calcium and Phosphorus of Patients Hemodialysis in Zahedan  

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Full Text Available Secondary hyperparathyroidism and its effects on bone tissue are among the most important complication of end-stage renal disease. In the present study, we investigated correlation between the serum parathormone level (PTH of hemodialysis men and women with calcium (Ca, phosphorus (Pi and alkaline phosphatase (ALP. We studied 30 chronic renal failure hemodialysis patients 16 men and 14 women, aged 22-66 years old (average 44 years old with dialysis duration of 5 months to 14 years. We measured the serum Ca., Pi and ALP in intervals of 3 months and serum PTH levels was measured in 3 month. Data analysis was performed using SPSS software. Our results showed that serum PTH and ALP levels were higher in women than in men (90% versus 70%, but abnormal range of serum Ca and Pi levels were higher in men then women (Ca : 8% versus 2%, Pi :58% versus 50%. Hemodialysis patients showed correlation between PTH and ALP (p<0.05, but the correlation of PTH with Ca and Pi levels was not statistically significant. No correlation was observed between PTH and ALP and Pi in men, however it was significant between PTH and Ca (p<0.08, r = - 0.63. The women showed correlation between PTH and ALP (p<0.05, but not between Ca and Pi levels. Based on the findings of this study, Secondary hyperparathyroidism and its effects on bone tissue were greater in women than men hemodialysis.

R. Saravani

2007-01-01

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Separation and quantification of corticosteroid-induced, bone and liver alkaline phosphatase isoenzymes in canine serum.  

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Quantifying alkaline phosphatase (ALP) isoenzymes in canine serum would provide a useful index in a clinical laboratory. To achieve this goal, we tested a semi-automatic assay combining wheat germ lectin (WGL) precipitation and chemical inhibition of isoenzymes of the TNS gene with levamisole to quantify bone ALP (BALP) and corticosteroid-induced ALP (CALP), respectively. The liver ALP (LALP) isoenzyme was then calculated from the equation: TALP = BALP + LALP + CALP BALP, LALP and CALP standards from serum of puppies, bile-duct ligated dogs and dogs on 4.4 mg/kg/day prednisolone for 30 days, respectively, were used. The suitability of standard sera was tested by affinity electrophoresis. Levamisole (4.2 mM) inhibits 98% of BALP and LALP but only 42% CALP. Multiplying measured CALP by 1.8 gives the total CALP value in serum. WGL precipitated 92.3% BALP, 23.3% LALP and 26.8% heated CALP standards. These values were used to adjust precipitated ALP to obtain the exact levels of BALP. WGL was then tested on pooled serum standards in which the relative proportions of all the ALPs were known and controlled. BALP was adequately quantified except when LALP and CALP levels were extremely high. The assay was also applicable under conditions resulting in high ALP. Therefore, combining WGL and levamisole inhibition provides an adequate separation and quantification of canine ALP isoenzymes. The method has great potential for diagnostic use and should be tested further for routine implementation. PMID:9465780

Syakalima, M; Takiguchi, M; Yasuda, J; Hashimoto, A

1997-12-01

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Serum Bone Alkaline Phosphatase in Assessing Illness Severity of Infected Neonates in the Neonatal Intensive Care Unit  

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Background: Infections can influence bone metabolism of neonates, which may lead to changes in some bone metabolism biomarkers. The purpose of this study was to determine whether serum bone alkaline phosphatase (BALP), osteocalcin (OC) and beta carboxy-terminal peptide of type I collagen (CTX), as specific biomarkers of bone metabolism, can be used to assess the severity of neonatal infections.

Zhang, Yaozong; Xue, Chenguang; Zhu, Tian; Du, Xiaolan; Su, Nan; Qi, Huabing; Yang, Jing; Shi, Yuan; Chen, Lin

2012-01-01

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Comparative Effects of Aluminium on Serum, Liver and Brain High and Low Molecular Weight Alkaline Phosphatase in Rats  

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Full Text Available The relationship between aluminium treatment and changes in the concentration of serum, liver and brain high- and low molecular weight alkaline phosphatase has been investigated in this manuscript. Results obtained showed that every other day intrapritoneally injection of 186 ?mol kg-1 of aluminium (AlCl3.6H2O, in male rats for 2 weeks resulted in decreasing the level of liver and brain alkaline phosphatase by 14.9 and 9.9%, respectively, whereas an elevation of serum levels of this enzyme by 21.1% was seen in comparison to untreated controls (p<0.05. Long-term exposure to 74.5 ?mol kg-1 of this salt, showed a statistically significant reduction in liver and brain levels of alkaline phosphatase by 15.8 and 12.3%, respectively and an increment in serum activity of the enzyme by 30.9% in compared to control group (p<0.05. Using gel filtration chromatography technique with sephacryl S300 showed that, in comparison to control groups, serum and liver homogenate from aluminium treated groups had a significant level of high molecular weight alkaline phosphatase, which might be considered as a potential biomarker for aluminium toxicity.

A.A. Moshtaghie

2006-01-01

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ALP (Alkaline Phosphatase) Test  

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... of this website will be limited. Search Help? ALP Share this page: Was this page helpful? Also ... Panel ; Bone Markers ; Alkaline Phosphatase Isoenzymes; Bone Specific ALP At a Glance Test Sample The Test Common ...

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Serum total and bone alkaline phosphatase levels and their correlation with serum minerals over the lifespan of sheep.  

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This study aimed to assess serum total alkaline phosphatase (ALP) and its bone isoform (BALP) levels during the ageing and in different physiologic states of sheep, in order to expand the knowledge about the variation of these biomarkers over the sheep lifespan. Ninety female sheep were divided into nine groups of various ages and physiological states (dry, lactation and pregnancy). Serum ALP, BALP and mineral levels were determined by commercial immunoassay, molecular absorbance spectrophotometry and chemical luminescence for BALP determination. Serum ALP and BALP decreased as sheep aged, and no statistically significant differences were obtained between ewes in different physiologic states. The continuous decline of serum BALP concentration along the sheep lifespan, namely in mature and old sheep, is a sign of decreasing bone turnover associated with ageing. Serum calcium concentrations increased slightly until 2 years of age and then showed a tenuous but statistically significant decrease in mature sheep, while serum phosphorus maintained an uninterrupted decrease as sheep matured. The knowledge of serum values of bone biomarkers throughout the sheep lifespan may be useful in preclinical orthopaedic research studies and for animal science studies using sheep. PMID:24334071

Sousa, Cristina P; Azevedo, Jorge T; Silva, Amélia M; Viegas, Carlos A; Reis, Rui L; Gomes, Manuela E; Dias, Isabel R

2014-06-01

 
 
 
 
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High serum alkaline phosphatase levels, a study in 181 Thai adult hospitalized patients  

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Full Text Available Abstract Background Alkaline phosphatase (ALP is an important enzyme mainly derived from the liver, bones and in lesser amounts from intestines, placenta, kidneys and leukocytes. An increase in ALP levels in the serum is frequently associated with a variety of diseases. This study was done in order to determine the diseases associated with a high ALP level among Thai adult hospitalized patients. Method A review was made of medical records of inpatients with high ALP level above 1000 IU/L in King Chulalongkorn Memorial Hospital, Thailand from January 1999 to December 1999. Excluded were cases of (a patients who have bone involvements with malignancies, (b pediatric patients younger than 15 years old and c HIV-seropositive patients. Results A total of 181 hospitalized patients with eligible medical records were identified (96 males and 85 females, mean age 49.4 ± 16.1 years. Their ALP levels ranging from 1,001 to 3,067 IU/L, these patients were divided into four groups. Conclusion High serum ALP levels in hospitalized patients were commonly found in three major groups having obstructive biliary diseases, infiltrative liver disease and sepsis. The study results were in accordance with previous reports in developed countries. Nonetheless, cholangiocarcionoma and some tropical diseases unique to our setting were also detected in these cases. where there was a marked elevation of serum ALP.

Wiwanitkit Viroj

2001-08-01

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Pesticide-induced Changes in Serum Levels of Acid Phosphatase, Alkaline Phosphates and Glutamate Oxaloacetate Transaminase in Rats  

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Full Text Available Three insecticides (Monitor, Talstar and Benzenehexachloride {BHC}, orally administered at the doses of 0.06, 0.5 and 0.2 mg kg 1 body weight respectively for 21 days affected the body weight and biochemical environment of blood and liver in the rats. Talstar treatment resulted in an increase in the body weight while Monitor and BHC treatment reduced the body weight. After 21 days of the Monitor treatment, activities of acid phosphatases increased (55% and alkaline phosphates decreased (2% in the serum while slight inhibition was observed in the serum glutamate oxaloacetate transaminase activity. Benzenehexachloride treatment resulted in increased serum levels of acid phosphatase, alkaline phosphatase and glutamate oxaloacetate transaminase (42, 2 and 18% respectively. Talstar treatment decreased the activities of acid phosphatase and alkaline phosphatase (76 and 5%, respectively while glutamate oxaloacetate transaminase activity was increased (25%. These data suggest that chronic exposure to Monitor, Talstar and BHC causes hepatocyte necrosis and increases the liver enzyme synthesis.

Muhammad Nasim Khan

2003-01-01

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Effect of Scoparia dulcis on Trypanosoma brucei Induced Alterations in Serum Transaminase, Alkaline Phosphatase and Bilirubin in the Rabbit  

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Full Text Available The present study reported the effect of S. dulcis on Trypanosome induced alterations in serum transaminases, Alkaline phosphatase and bilirubin in the rabbit. There were significant increases in Alkaline Phosphatase (ALP, Serum Glutamic Oxaloacetic Transaminase (SGOT, Serum Glutamic-Pyruvic Transaminase (SGPT, total bilirubin (T. Bil. and conjugated bilirubin (C. Bil. in infected animals relative to control. The values obtained for infected animals that were treated with Scoparia dulcis at a daily oral dose of 12.5 mg kg-1 body weight compared well with controls and were significantly lower than those observed for infected but untreated animals. These results suggest that S. dulcis effectively resists these Trypanosome induced changes in the rabbit.

N.E.J. Orhue

2004-01-01

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Alkaline phosphatase: an overview.  

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Alkaline phosphatase (ALP; E.C.3.I.3.1.) is an ubiquitous membrane-bound glycoprotein that catalyzes the hydrolysis of phosphate monoesters at basic pH values. Alkaline phosphatase is divided into four isozymes depending upon the site of tissue expression that are Intestinal ALP, Placental ALP, Germ cell ALP and tissue nonspecific alkaline phosphatase or liver/bone/kidney (L/B/K) ALP. The intestinal and placental ALP loci are located near the end of long arm of chromosome 2 and L/B/K ALP is located near the end of the short arm of chromosome 1. Although ALPs are present in many mammalian tissues and have been studied for the last several years still little is known about them. The bone isoenzyme may be involved in mammalian bone calcification and the intestinal isoenzyme is thought to play a role in the transport of phosphate into epithelial cells of the intestine. In this review, we tried to provide an overview about the various forms, structure and functions of alkaline phosphatase with special focus on liver/bone/kidney alkaline phosphatase. PMID:24966474

Sharma, Ujjawal; Pal, Deeksha; Prasad, Rajendra

2014-07-01

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Isozyme profile and tissue-origin of alkaline phosphatases in mouse serum.  

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Mouse serum alkaline phosphatase (ALP) is frequently measured and interpreted in mammalian bone research. However, little is known about the circulating ALPs in mice and their relation to human ALP isozymes and isoforms. Mouse ALP was extracted from liver, kidney, intestine, and bone from vertebra, femur and calvaria tissues. Serum from mixed strains of wild-type (WT) mice and from individual ALP knockout strains were investigated, i.e., Alpl(-/-) (a.k.a. Akp2 encoding tissue-nonspecific ALP or TNALP), Akp3(-/-) (encoding duodenum-specific intestinal ALP or dIALP), and Alpi(-/-) (a.k.a. Akp6 encoding global intestinal ALP or gIALP). The ALP isozymes and isoforms were identified by various techniques and quantified by high-performance liquid chromatography. Results from the WT and knockout mouse models revealed identical bone-specific ALP isoforms (B/I, B1, and B2) as found in human serum, but in addition mouse serum contains the B1x isoform only detected earlier in patients with chronic kidney disease and in human bone tissue. The two murine intestinal isozymes, dIALP and gIALP, were also identified in mouse serum. All four bone-specific ALP isoforms (B/I, B1x, B1, and B2) were identified in mouse bones, in good correspondence with those found in human bones. All mouse tissues, except liver and colon, contained significant ALP activities. This is a notable difference as human liver contains vast amounts of ALP. Histochemical staining, Northern and Western blot analyses confirmed undetectable ALP expression in liver tissue. ALP activity staining showed some positive staining in the bile canaliculi for BALB/c and FVB/N WT mice, but not in C57Bl/6 and ICR mice. Taken together, while the main source of ALP in human serum originates from bone and liver, and a small fraction from intestine (<5%), mouse serum consists mostly of bone ALP, including all four isoforms, B/I, B1x, B1, and B2, and two intestinal ALP isozymes dIALP and gIALP. We suggest that the genetic nomenclature for the Alpl gene in mice (i.e., ALP liver) should be reconsidered since murine liver has undetectable amounts of ALP activity. These findings should pave the way for the development of user-friendly assays measuring circulating bone-specific ALP in mouse models used in bone and mineral research. PMID:23313280

Halling Linder, Cecilia; Englund, Ulrika H; Narisawa, Sonoko; Millán, José Luis; Magnusson, Per

2013-04-01

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Serum alkaline phosphatase as a diagnostic criterion in experimental radiation disease  

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In the serum of full grown male Wistar rats irradiated with 60Co (100, 30a, 500 and 800 R) and in a control group the activity of serum alkaline phosphatase and its isoenzymes was investigated at various time intervals after irradiation. Changes in the general activity had a phasic character. A statistically significant drop of activity was found between the 2nd and 5th day after 500 R and permanently from the 2nd day after irradiation with 800 R. Changes in the activity of the intestinal and liver isoenzyme also had a phasic character. The activity of the intestinal isoenzyme declined significantly already on the first day after irradiation with 300 R and higher doses; the drop on the 2nd day after irradiation with 100 R was significant at the 5% level of significance. Investigation of the activity of the intestinal isoenzyme can be used as a rough biological dosimeter. The serum activity of the hepatic isoenzyme declined significantly on the 3rd to 5th day after irradiation with 500 and 800 R, which is in keeping with changes of the enzyme activity in the liver. After a dose of 800 R a progressing decline of activity of the bone isoenzyme was found even after 100 and 500 R. The findings are typical for the development of the gastrointestinal and osseous radiation syndrome. They may be used for the evaluation of the effect of radioprotective substances and therapeutic procedures and as a simple biochemical diagnostic and prognostic criterion of the action of ionizing radiation in experiments on rats. (author)

1976-01-01

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Effect of Scoparia dulcis on Trypanosoma brucei Induced Alterations in Serum Transaminase, Alkaline Phosphatase and Bilirubin in the Rabbit  

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The present study reported the effect of S. dulcis on Trypanosome induced alterations in serum transaminases, Alkaline phosphatase and bilirubin in the rabbit. There were significant increases in Alkaline Phosphatase (ALP), Serum Glutamic Oxaloacetic Transaminase (SGOT), Serum Glutamic-Pyruvic Transaminase (SGPT), total bilirubin (T. Bil.) and conjugated bilirubin (C. Bil.) in infected animals relative to control. The values obtained for infected animals that were treated with Scopa...

2004-01-01

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Serum alkaline phosphatase (SAP) activity following exposure to cadmium and/or 60Co gamma irradiation  

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Two hundred and sixteen male Sprague-Dawley (S-D) rats were assigned at random to nine groups of 24 rats each. Rats were injected with cadmium (Cd) intraperitoneally every 3 days for 29 days for a total of nine injections. Injection doses were 0, 1.0, or 2.5 mg Cd kg-1 body weight. Twenty-four hours after the last Cd injection (day 30), each rat received an acute whole-body 60Co gamma radiation dose of 0, 3.62, or 5.43 Gray (Gy) at a dose rate of 3.304 Gy min-1. Eight rats from each of 9 groups were sacrificed on day 1, 7, or 21. High dose radiation administered 24 hours following the last dosage of Cd caused significantly elevated serum alkaline phosphatase levels, whereas high dose cadmium caused the enzyme to be significantly depressed. When Cd and radiation were used as the co-insult, the combination of high Cd-high radiation was more effective than either cadmium or radiation alone, suggesting a previously reported cadmium metal protection against the radiation. Although the precise mechanism is unknown, they speculate that the protection afforded by cadmium against radiation might be attributed to different conformations of metal-induced metallothionein cysteine clusters. Further, these clusters are likely dependent upon conversion between conformational forms requiring specific levels of metal ion site occupancy

1987-01-01

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Pre-operative Serum Alkaline Phosphatase as a Predictor for Hypocalcemia Post-Parathyroid Adenectomy  

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Full Text Available Background. Post-operative hypocalcemia (POH may complicate parathyroidectomy for primary hyperparathyroidism. This study investigates the relationship between POH and pre-operative risk factors to identify a simple method to predict POH risk.Methods. Retrospective data on risk factors for 29 patients was collected for age, pre-operative serum calcium, alkaline phosphatase (ALP, parathyroid hormone (PTH, adenoma size, gender, and bisphosphonate pre-treatment. These were screened to exclude those with small effect sizes, and analyzed using Univariate General Linear Modeling (GLM with trough serum calcium (TSC as the dependent variable. The regression function of the significant variables against TSC was plotted with 95% CI fit lines. The cut-off regression value was read from the lower fit line for the threshold TSC of 2.0 mmol/L.Results. After screening, log-transformed age (r=0.600, ALP (r=-0.415, and PTH (r=-0.433 were entered into GLM analysis, which showed that only ALP was significant (p=0.016 Eta-squared=0.220. The GLM model had a partial Eta-squared of 0.559 with 98% observed power. The plot of TSC against log-ALP gave an ALP cut-off of 340 U/L.Conclusions. The study shows that there is a strong relationship between ALP and TSC, and that patients with a pre-operative ALP less than 340 U/L are unlikely to have symptomatic POH (100% sensitivity, 95% specificity. While vitamin D was not analyzed in this study, the ALP cut-off is conservative and should still screen out cases with severe vitamin D deficiency. We therefore recommend that pre-operative ALP be utilized to complement clinical protocols for POH management in parathyroid adenomectomy patients.

Seng Cheong Loke, Alvin Wai Kit Tan, Rinkoo Dalan, Melvin Khee-Shing Leow

2012-01-01

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Serum alkaline phosphatase activity is not a marker for neoplastic transformation of esophageal nodules in canine spirocercosis  

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BACKGROUND: Spirocerca lupi is a nematode of Canidae that matures within the esophageal wall to form fibroblastic nodules with potential for malignant transformation. Diagnosis is based on histopathologic examination, but false-negative results may be obtained from samples collected by endoscopy. Serum alkaline phosphatase (ALP) activity, frequently increased in hepatobiliary disease, is also increased in a variety of neoplastic conditions in dogs, including appendicular ost...

Mukorera, Varaidzo; Merwe, Liesel L.; Lavy, Eran; Aroch, Itamar; Dvir, Eran

2011-01-01

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The isoelectric focusing properties of serum alkaline phosphatase in disease and following prednisolone and phenylbutazone administration in the horse.  

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This study was undertaken to ascertain if the isoelectric focusing pattern of serum alkaline phosphatase (AP) from sick horses with high activity is useful for determining its tissue origin. The effect of oral prednisolone and phenylbutazone therapy on this enzyme in healthy horses was also investigated. The sick horses were divided into three groups: hepatic, intestinal and miscellaneous. All sera had approximately thirteen bands of AP activity when focused on agarose gels with a pH gradient...

Ellison, R. S.; Jacobs, R. M.

1990-01-01

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Assessment of the serum levels of bone alkaline phosphatase with a new immunoradiometric assay in patients with metabolic bone disease  

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The authors measured serum bone alkaline phosphatase (B-ALP) with a new immunoradiometric assay (IRMA) in a large sample of healthy controls comprising 173 women and 180 men, 20-88 yr of age, and in patients with metabolic bone disease. Using serum samples from patients with liver disease and patients with Paget's disease with elevated total alkaline phosphatase (T-ALP) as a source of, respectively, liver and bone isoenyzmes, they determined a liver cross-reactivity of the IRMA of 16% that was confirmed by electrophoresis of the circulating alkaline phosphatase isoenzymes. The IRMA was linear for serial sample dilutions, the recovery ranged from 89-110%, and the intra- and interassay variations were below 7% and 9%, respectively. B-ALP increased linearly with age in both sexes, and the mean B-ALP serum levels were not significantly different for women and men (11.3 ± 4.8 ng/mL for women; 11.0 ± 4.0 ng/mL for men). The increase in B-ALP after the menopause was significantly higher than that in T-ALP (+77% vs. +24%; P<0.001). When the values of postmenopausal women were expressed as the SD from the mean of premenopausal women, the mean Z scores were 2.2± 1.8 for B-ALP and 0.9 ± 1.3 for T-ALP (P<0.001 between the two)

1993-10-01

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Serum Proteins, Thyroid Hormones and Alkaline Phosphatase Concentrationsin Acute Experimental Trypanosoma congolense Infection in Yankasa Sheep Immunomodulated with Levamisole  

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Full Text Available Serum proteins, thyroid hormones and alkaline phosphatase concentrations were measured in Yankasa sheep experimentally infected with T. congolense. Parasitemia occurred in the T. congolense infected sheep immunomodulated with levamisole two days earlier than the infected group without immunodulation.Packed cell volume decreased significantly(p0.05 in the infected sheep with and without immunomodulation when compared to the controls throughout the period of the experiment. In general, levamisole administration did not appear to alter the infection when compared to the infected group without immunomodulation.

I.A. Lawal

2007-01-01

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Determination of the Relationship between Serum Calcium, Phosphor and Alkaline Phosphatase with Peripheral Giant Cell Granuloma in Iranian Patients  

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Peripheral giant cell granuloma (P.G.C.G) is an exophytic lesion with an approximate size of"n0.5-1.5 mm. It usually occurs on gingival and alveolar ridge of mandible particularly in molar and"npremolar region. The relation of serum calcium (Ca), Phosphor (P), and alkaline phosphatase (Alk) levels"nto P.G.C.G is yet controversial. In this descriptive study, 33 patients with P.G.C.G were chosen and"nserum Ca, P, and Alk levels compared with the normal range. In all patients...

Saheb-Jamei M; Hr,  abdossamadi

2000-01-01

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Influence of radioprotective agents on the activities of isoenzymes of blood serum alkaline phosphatase in irradiated dogs  

International Nuclear Information System (INIS)

Changes of the total serum activity of alkaline phosphatase (AP) and its bone and intestinal isoenzymes were studied in dogs ?-irradiated by 3.0 Gy. The activities of AP isoenzymes were determined by means of a heat inactivation-inhibition method. After irradiation the serum AP activities were lower in general. In case of the bone isoenzyme the decrease was most pronounced. The changes of the total AP activity and of the intestinal isoenzyme were less distinct. The i.m. administration of the radioprotective mixture cystamine with mexamine (24 mg/kg + 4 mg/kg) before irradiation led only to a moderate reduction of the radiation-altered AP values in the dogs. The present results indicate that the radioprotective way used here renders only an unsatisfactory protection for large laboratory animals against ionizing radiation. (orig.)

1980-01-01

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Application of ascorbic acid 2-phosphate as a new voltammetric substrate for alkaline phosphatase determination in human serum  

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Full Text Available An electrochemical assay of the enzyme alkaline phosphatase (ALP using ascorbic acid 2-phosphate (AAP as a new voltammetric substrate has been described in this paper. In the alkaline buffer solution the ALP enzymatic hydrolysis product of AAP was ascorbic acid (AA, which was an electro-active substance and had a sensitive differential pulse voltammetric (DPV oxidative response on glassy carbon electrode (GCE at +380 mV (versus Ag/AgCl, so the activity of ALP could be monitored voltammetrically of the oxidative peak current of AA. The electrochemical behaviours of AA were carefully studied and the AA standard solution could be measured by DPV method in the linear range from 10.0 to 1000.0 ?mol/L with the detection limit of 8.0 ?mol/L. The optimal conditions for ALP enzymatic reaction and the voltammetric detection were optimized. Under the optimal conditions the calibration curve for ALP assay exhibited a linear range from 0.4 to 2000.0 U/L with a detection limit of 0.3 U/L. This proposed method was further applied to determine the ALP content in healthy human serum and the results were in good agreement with the traditional p-nitrophenyl phosphate spectrophotometric method. The kinetic constants of enzymatic reaction were also investigated with the apparent kinetic constant Km as 2.77 mmol/L and the maximum velocity Vmax as 0.33 mol/min.

Wei Sun

2005-12-01

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Prognostic significance of serum alkaline phosphatase in osteosarcoma of the extremity treated with neoadjuvant chemotherapy: recent experience at Rizzoli Institute.  

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In 560 patients with high-grade osteosarcoma of the extremity treated with 5 different protocols of neoadjuvant chemotherapy at a single institution between 1983 and 1995, the pre-treatment serum alkaline phosphatase (SAP) was examined to evaluate whether the enzyme levels had a clinical value in predicting the course of the disease. SAP was normal in 302 (54%) patients and high in 258 (46%). High levels of SAP was observed significantly and independently more frequently in male patients over 14-years-old, and in tumours larger than 150 ml and of osteoblastic subtypes. The 5-year event-free survival (EFS) and overall survival (OS) for all patients were respectively 60 and 68%. With multivariate analysis only two factors were independently correlated with the 5-year EFS: SAP levels (p=0.002) and the grade of chemotherapy-induced necrosis (p=0.0001). The authors conclude that in planning randomized clinical trials of neoadjuvant treatment for osteosarcoma, patients should be stratified according to SAP levels, and that when tailoring the aggressiveness of postoperative chemotherapy to the risk of relapse, in addition to the histologic response to preoperative treatment, the SAP levels should also be considered. PMID:11748477

Bacci, Gaetano; Longhi, Alessandra; Ferrari, Stefano; Lari, Stefano; Manfrini, Marco; Donati, Davide; Forni, Cristiana; Versari, Michela

2002-01-01

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Alkaline phosphatase and transaminase activity in rat liver and blood serum at delayed periods following external ?-irradiation combined with internal exposure to plutonium 239  

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A study was made of activity of alkaline phosphatase and alanine- and aspartate aminotransferase in rat liver and blood serum at remote times after external ?-irradiation combined with internal exposure to 239Pu nitrate delivered in two chronically effective doses. The radionuclide was shown to be mainly responsible for the changes observed in activity of the enzymes under study. The degree to which the changes were manifest depended upon dose of plutonium administered

1986-01-01

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High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China  

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Abstract Background Osteosarcoma is a malignant tumor with high ability to form invasion and metastasis. Identifying prognostic factor in osteosarcoma is helpful to select those patients for more aggressive management. Our study evaluated serum alkaline phosphatase (ALP) cooperating with matrix metalloproteinase-9 (MMP-9) as an important prognostic predictor for local recurrence and distant metastasis of osteosarcoma. Methods 177 cases were included from the ost...

Han Ju; Yong Bicheng; Luo Canqiao; Tan Pingxian; Peng Tingsheng; Shen Jingnan

2012-01-01

40

Pesticide-induced Changes in Serum Levels of Acid Phosphatase, Alkaline Phosphates and Glutamate Oxaloacetate Transaminase in Rats  

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Three insecticides (Monitor, Talstar and Benzenehexachloride {BHC}), orally administered at the doses of 0.06, 0.5 and 0.2 mg kg 1 body weight respectively for 21 days affected the body weight and biochemical environment of blood and liver in the rats. Talstar treatment resulted in an increase in the body weight while Monitor and BHC treatment reduced the body weight. After 21 days of the Monitor treatment, activities of acid phosphatases increased (55%) and alkaline phosphates dec...

Muhammad Nasim Khan; Tahira Sarwar

2003-01-01

 
 
 
 
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Isoenzimas de fosfatasa alcalina en el suero de pacientes con insuficiencia renal / Alkaline phosphatase isoenzymes in the serum of patients with renal insufficiency  

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Full Text Available SciELO Spain | Language: Spanish Abstract in spanish Objetivo: estudiar la utilidad clínica de la determinación sérica de las isoenzimas de fosfatasa alcalina en pacientes con insuficiencia renal. Material y métodos: se midieron las isoenzimas de fosfatasa alcalina en un grupo de 58 pacientes: 22 con insuficiencia renal aguda (IRA) y 36 con fallo rena [...] l crónico (IRC) y sometidos a hemodiálisis, comparándose los resultados con los de una población de 30 adultos sanos. Las isoenzimas intestinal, ósea, hepática, macromolecular e intestinal variante se separaron por electroforesis sobre gel de agarosa, cuantificándose por densitometría. Resultados: la actividad total de fosfatasa alcalina se mostró significativamente aumentada en ambos grupos patológicos (p Abstract in english Objective: The aim of this study was to test the utility of serum alkaline phosphatase isoenzymes determination from patients with renal insufficiency. Material and methods: serum levels of alkaline phosphatase isoenzymes were determined in a group of 58 patients: 22 of them suffering acute renal in [...] sufficiency (ARI) and 36 with chronic renal failure (CRF) undergoing regular hemodialysis, results obtained were compared from a population of 30 healthy adults. Intestinal, bone, liver, macromolecular and intestinal variant isoenzymes, were separated by electrophoresis on agarose gel and quantified using a densitometer. Results: were found a significant increase the total alkaline phosphatase activity in both pathologic groups (p

Sánchez Navarro, Mª. R.; Fernández-Conde, Mª. E.; Blanco Martín, S.; Samaniego, C..

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Alkaline phosphatase: Distinguishing between tuberculous and nontuberculous pleural effusion  

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Full Text Available Objectives: To evaluate the value of pleural fluid alkaline phosphatase and pleural fluid/serum alkaline phosphatase ratio for the purpose of differentiating tuberculous from nontuberculous pleural effusion. Materials and Methods: A total of 60 indoor patients, admitted to our hospital, having pleural effusion and suffering from varying etiologies, were included in this study. According to the final diagnosis, these 60 patients were divided into two groups: Tuberculous (30 and nontuberculous (30 pleural effusion. Results: The mean pleural alkaline phosphatase and pleural fluid/serum alkaline phosphatase ratio was significantly higher in tuberculous compared to nontuberculous pleural effusion. ( P < 0.0001. In receiver operating characteristic curve analysis, sensitivity and specificity values were 90% and 80% for a cut-off value of 71 IU/L for pleural alkaline phosphatase activity; and were 90% and 86.66% for a cut-off value of 0.51 for pleural fluid/serum alkaline phosphatase ratio. Conclusion: From this study it is concluded that alkaline phosphatase activity remains a useful test in differentiation of tuberculous from nontuberculous pleural effusion.

Jadhav Ashish

2009-01-01

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Alkaline phosphatase: Distinguishing between tuberculous and nontuberculous pleural effusion  

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Objectives: To evaluate the value of pleural fluid alkaline phosphatase and pleural fluid/serum alkaline phosphatase ratio for the purpose of differentiating tuberculous from nontuberculous pleural effusion. Materials and Methods: A total of 60 indoor patients, admitted to our hospital, having pleural effusion and suffering from varying etiologies, were included in this study. According to the final diagnosis, these 60 patients were divided into two groups: Tuberculous (30) a...

2009-01-01

44

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

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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 1?g 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.

Luiz Augusto de Souza

2011-12-01

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Relationship between serum heat-stable alkaline phosphatase activity and blood pressure in patients with pre-eclampsia and eclampsia  

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Full Text Available Background : The objective of this study was to explore the relationship, if any, between theserum heat-stable alkaline phosphatase (HS-ALP activity and the blood pressure (BP of patients with pre-eclampsia and eclampsia. Method : The activity of HS-ALP was measured using the 4– nitrophenyl phosphate (4– NPP method after incubation at a high temperature of 65 0 C for exactly 30 minutes in one hundred normal pregnant women and in another one hundred with pre-eclampsia/eclampsia. The normal pregnant women were used as controls. The blood pressure (BP, systolic as well as diastolic was measured in each of the studied patient using desktop mercury sphygmomanometer. Results :In the patients with pre-eclampsia/eclampsia, it was found that the higher the systolic and diastolic BP, the higher is the activity of the HS-ALP. Conclusion : It can be concluded that the HS-ALP activity in patients with pre-eclampsia/eclampsia is positively related to the severity of the hypertension and therefore this could help in detecting early complication.

Aliyu I

2006-03-01

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Effects of "6"0Co gamma-ray local irradiation on rat liver on alkaline phosphatase, lactate dehydrogenase and catalase in the liver and serum  

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Rats were given a single exposure of various doses (0, 5, 50, 500, and 5000 rads) to local irradiation of "6"0Co ?-ray on liver. Activities of alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and catalase in the serum and liver were measured at various time intervals after irradiation. These results were summarized as follows; 1. ALP activity in the serum had no effect on irradiation up to 500 rads, but in the case of 5000 rads irradiation exhibited a marked loss from 4 days after irradiation. ALP activity in the liver to 5000 rads exposure on 7 days after irradiation increased, on the other hand in the serum decreased, and the patterns of ALP activities in the liver and serum to the irradiation doses were opposite. 2. LDH activity in the serum by exposure to 5, 500 and 5000 rads increased at 4 days after irradiation, but at 7 days significantly decreased. LDH activity in the liver to the irradiation doses on 7 days after irradiation did not markedly change, but in the serum it tended to be low in inverse proportion to the irradiation doses. 3. Catalase activity in the serum to 50 and 500 rads exposure increased at 4 days after irradiation and decreased at 7 days, but to 5000 rads exposure it decreased in the course of time. Catalase activity in the liver and serum on 7 days after irradiation were inversely proportional to irradiation doses. It is difficult that catalase activity makes a index of clinical irradiation effects, because catalase activity decrease under the various conditions, such as cancer, anemia, infection of bacterias and so on. Since activities of ALP and LDH increase in almost disease, decrease of ALP activity and decrease following temporary increase of LDH activity by irradiation may be able to become a clinical indicator on irradiation effects. (author)

1980-01-01

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The Role of Whole-Body FDG PET/CT, Tc 99m MDP Bone Scintigraphy, and Serum Alkaline Phosphatase in Detecting Bone Metastasis in Patients with Newly Diagnosed Lung Cancer  

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Bone scan (BS) and serum alkaline phosphatase (ALP) concentration are used to detect bone metastasis in malignancy, although whole-body fluoro-D-glucose positron emission tomography computed tomography (FDG PET/CT) is being used increasingly. But BS is still used for the detection of metastatic bone lesion. So we compared the usefulness of PET/CT, BS, and serum ALP in detecting bone metastases in patients with newly diagnosed lung cancer. The medical record database was queried to identify al...

Min, Joo-won; Um, Sang-won; Yim, Jae-jun; Yoo, Chul-gyu; Han, Sung Koo; Shim, Young-soo; Kim, Young Whan

2009-01-01

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Hepatic uptake of intestinal alkaline phosphatase. A morphological and kinetic study in the rat  

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The mucosa of the small intestine is extremely rich in the enzyme alkaline phosphatase. A considerable amount of this intestinal enzyme enters the general circulation via the thoracic duct. However, in man none or only a small amount of intestinal alkaline phosphatase is present in the serum, suggesting that this enzyme is efficiently eliminated from the circulation. A rapid removal of injected intestinal alkaline phosphatase from the circulation has been demonstrated in dog, cat and rat. How...

1980-01-01

49

High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China  

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Full Text Available Abstract Background Osteosarcoma is a malignant tumor with high ability to form invasion and metastasis. Identifying prognostic factor in osteosarcoma is helpful to select those patients for more aggressive management. Our study evaluated serum alkaline phosphatase (ALP cooperating with matrix metalloproteinase-9 (MMP-9 as an important prognostic predictor for local recurrence and distant metastasis of osteosarcoma. Methods 177 cases were included from the osteosarcoma patients treated at 1st Affiliated Hospital of Sun Yat-sen University (1999-2008. Pre-chemotherapy serum ALP (pre-ALP were studied and correlated with tumor recurrence, lung metastasis and patient survival. MMP-9 protein in tumor tissues was detected by immunohistochemistry and correlated with pre-ALP level. Results Pre-ALP were partitioned into normal, high, and very high groups, in each group the incidence of metastases was 12.2%, 21.2% and 34.6%, respectively (p = 0.007. In the three groups the mean disease-free survival (DFS was 57 ± 3.15, 28 ± 3.57 and 14 ± 3.35 months, respectively (p Conclusions Pre-ALP was an independent prognostic factor for the survival of osteosarcoma patients in south China, and correlated with MMP-9 expression and lung metastasis. ALP can also serve as a prognostic marker for treatment, and merit large-scale validation studies.

Han Ju

2012-02-01

50

Transient increase in total serum alkaline phosphatase predicts radiological response to systemic therapy in breast cancer patients with osteolytic and mixed bone metastases.  

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Assessment of the response of bone metastases to endocrine or chemotherapy is difficult, and true response rate is probably underestimated by UICC criteria. Biochemical markers of osteoblast activity, which is linked with bone healing, could be useful for early detection of treatment response. We studied changes in osteoblast function, assessed by serial serum total alkaline phosphatase (tALP) at 0, 1, 2, 3 months from the start of systemic therapy, in 31 patients bearing bone metastases from advanced breast cancer. After 1 month of endocrine or cytotoxic treatment, all responding patients (7/31) showed a significant rise in tALP (mean increase: 351 IU/l, p < 0.05) followed by a gradual decrease over the subsequent months (tALP flare). Transient increase in tALP was also found in 2/16 patients with stable disease who benefited from therapy. 2/8 patients with progressive disease showed a rise indistinguishable from responders, but the subsequent decrease was not apparent. These observations suggest that serum tALP profile is an earlier predictor of response than X-rays. PMID:8497373

Berruti, A; Osella, G; Raucci, C A; Roncari, A; Dogliotti, L

1993-01-01

51

Activity of alkaline phosphatase and its fractions in blood serum of rats with experimental hyperthyroidism, kept in a radionuclide contaminated area  

International Nuclear Information System (INIS)

In studies on male rats, with experimentally induced thyrotoxicosis a modifying effect of the stay in radioactively contaminated zone on the total alkaline phosphatase activity was shown. The data indicate disturbances in calcium-phosphoric metabolism, resulting in pronounced deviations from the normal range in the activity of thermolabile (bone) fraction (authors)

2002-01-01

52

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

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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 correlation between CXCR4 expression and serum ALP.Conclusion: CXCR4 siRNA/dextran-spermine nanoparticles appear to be highly effective, and may be suitable for further in vivo applications. Further research evaluation will be needed to determine the effect of CXCR4 silencing on serum ALP levels, which may be a useful marker to predict liver metastasis in colorectal cancer.Keywords: nanoparticles, cationized dextran, colorectal cancer, serum ALP enzyme, CXCR4, siRNA

Abedini F

2012-07-01

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Osteoblastic flare assessed by serum alkaline phosphatase activity is an index of short duration of response in prostate cancer patients with bone metastases submitted to systemic therapy.Gruppo Onco Urologico Piemontese (G.O.U.P).  

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A transient rise in serum alkaline phosphatase (ALP) activity (ALP flare) after androgen deprivation in prostate cancer patients with bone metastases has been previously correlated with both response to therapy and poor prognosis. In the present study we analyzed data coming from an Italian multicenter phase III, trial aimed to compare the efficacy of treatment with goserelin alone with that of goserelin plus mitomycin C. Sixty-seven bone metastatic patients were enrolled: 32 were treated wit...

Angeli, Alberto; Rocca Rossetti, Salvatore; Dogliotti, Luigi; Fontana, Dario; Berruti, Alfredo

1997-01-01

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Radioimmunoassay of human intestinal alkaline phosphatase  

International Nuclear Information System (INIS)

A new method of radioimmunoassay using the double antibody method for human intestinal alkaline phosphatase (ALP) was first elaborated. The following results were obtained: 1) In this system, the optimal antibody concentration is 10,000 times the dilution of the original anti-serum, and the optimal assay range is 0.5 to 25 ng. Enzymatic activity of 1 ng intestinal ALP is 4.1 King-Armstrong units. 2) In this system, the sera including intestinal ALP are divided to two groups. One group shows a dose response curve similar to that of purified intestinal ALP, and the other shows a lesser one. This reason is not clear. Hepatic ALP, osseous ALP and placental ALP in the sera show no response in this system. 3) In this system, the B/T value of 50 ?g of purified human placental ALP is almost equal to 1 ng of purified human intestinal ALP. Similarly, the B/T value of 50 ?g of purified human intestinal ALP is equal to almost 5 ng of purified human placental ALP. This shows that cross-reaction exists between intestinal and placental ALPs at high concentrations. (J.P.N.)

1976-01-01

55

Inhibition of alkaline phosphatase activity by glucose.  

Science.gov (United States)

Non-enzymatic glycosylation (NEG) of alkaline phosphatase (AP) was studied after short- and long-term incubation with glucose and other carbohydrates. Glucose and amino sugars clearly inhibited the enzyme activity; this was in contrast to reducing and non-reducing disaccharides, which had an enhancing effect. After AP had been incubated with 18 nmol/l glucose for 180 minutes (short-term incubation), a subsequent extensive dialysis revealed full recovery of the enzymatic activity. This, plus the demonstration of a [3H]sodium borohydride-reducible glucose-protein adduct, indicated that initially a labile aldimine (Schiff base) had been formed. Binding experiments with [14C]glucose and failure of dialysis to achieve a recovery of enzymatic activity after long-term incubation suggested that subsequently a stable ketoamine product had been formed. This was further confirmed by the thiobarbituric acid test, which revealed 0.65 nmol 5-hydroxymethylfurfural/mg protein for glycosylated AP compared to 0.11 for the non-glycosylated control. Preliminary results further suggest that NEG of AP also occurs in vivo. Streptozotocin diabetic rats had significantly lower serum AP activities than did non-diabetic controls (mean +/- SD: 153.7 +/- 28.4 vs. 760.5 +/- 95.7 U/l; p less than 0.001). Blood glucose levels and serum AP activity, which had been determined simultaneously during an oral glucose tolerance test, showed without exception an inverse relationship in each of 32 healthy children studied. The biological significance of these findings remains to be established. PMID:6627674

Pollak, A; Coradello, H; Leban, J; Maxa, E; Sternberg, M; Widhalm, K; Lubec, G

1983-09-15

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Radiation inactivation of bovine intestinal alkaline phosphatase  

Science.gov (United States)

The effects of 60Co ?-radiation on aqueous solutions of alkaline phosphatase have been studied. The primary radicals of water radiolysis, e -aq, OH· and H· all contribute to the observed inactivation with inactivating efficiencies of 0.019, 0.019 and 0.04, respectively; O -2 also causes inactivation (efficiency = 0.014). The radical anions (SCN) -2, (Br) -2 and (I) -2 cause inactivation at neutral pH and evidence is presented that cysteine and histidine residues are sites for radical anion reaction. Kinetic evidence suggests that inactivation is due to general denaturation of the protein, rather than destruction of the substrate binding site. Fractionation of the irradiated solution using FPLC following by analysis using fluorescence spectroscopy suggests that one process which leads to inactivation is the formation of alkaline phosphatase dimerized via tyrosine residues.

Hasan, N. M.; McCall, P. R.; Moore, J. S.; Power, D. M.

1994-03-01

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Radiation inactivation of bovine intestinal alkaline phosphatase  

International Nuclear Information System (INIS)

The effects of 60CO ?-radiation on aqueous solutions of alkaline phosphatase have been studied. The primary radicals of water radiolysis, eaq-, OH. and H. all contribute to the observed inactivation with inactivating efficiencies of 0.019, 0.019 and 0.04, respectively; O2- also causes inactivation (efficiency = 0.014). The radical anions (SCN)2-, (Br)2-, and (I)2- cause inactivation at neutral pH and evidence is presented that cysteine and histidine residues are sites for radical anion reaction. Kinetic evidence suggests that inactivation is due to general denaturation of the protein, rather than destruction of the substrate binding site. Fractionation of the irradiated solution using FPLC following by analysis using fluorescence spectroscopy suggests that one process which leads to inactivation is the formation of alkaline phosphatase dimerized via tyrosine residues. (author)

1994-03-01

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Alkaline phosphatase as a periodontal disease marker  

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Background: The potential of alkaline phosphatase (ALP) as an important diagnostic marker of gingival crevicular fluid (GCF) has been the subject to investigation since 1970. ALP is stored in specific granules and secretory vesicles of the neutrophils and is mainly released during their migration to the site of infection. It is also present in bacteria within dental plaque, osteoblasts and fibroblasts. It has, thus, become important to elucidate whether GCF levels of ALP are potential ...

Malhotra Ranjan; Grover Vishakha; Kapoor Anoop; Kapur Rupika

2010-01-01

59

The serum levels of Receptor Activator of Nuclear Factor-?B Ligand, bone-specific alkaline phosphatase, osteocalcin and osteoprotegerin do not correlate with the radiographically assessed severity of idiopathic hip and knee osteoarthritis.  

Science.gov (United States)

OBJECTIVES: Determination of the serum levels of Receptor Activator of Nuclear Factor-?b Ligand, bone-specific alkaline phosphatase, osteocalcin and osteoprotegerin in patients suffering from osteoarthritis of varying severity and healthy controls and correlation of these results with the patients' age and the radiographically assessed severity of the disease. DESIGN AND METHODS: Patients suffering from hip (n=58) or knee (n=117) osteoarthritis and matched controls (n=19) were enrolled in this study. Patients underwent physical examination and standard radiographic evaluation before blood sampling. RESULTS: The serum levels of osteoprotegerin were positively correlated with age in all groups, whereas those of osteocalcin in the 'knee' group only. Osteoarthritis' severity and location did not have a statistically significant impact on the mean serum level of any marker in both groups. CONCLUSIONS: Based on our results, none of the studied markers can serve as a surrogate for radiographic imaging in patients suffering from hip and knee osteoarthritis. PMID:20951121

Kenanidis, Eustathios ?; Potoupnis, Michael E; Papavasiliou, Kyriakos A; Pellios, Stauros; Sayegh, Fares E; Petsatodis, George E; Karatzas, Nikolaos; Kapetanos, George A

2011-02-01

60

Studies on Alkaline Phosphatase Activity in Amphibians : II. Changes in Alkaline Phosphatase Activity during Development in Rana catesbeiana  

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The changes in alkaline phosphatase activity during development in Rana catesbeiana were examined by electrophoresis and spectrophotometry in embryos and hatching and hatched tadpoles, and in various visceral organs, such as the liver, pancreas, kidney and intestine of tadpoles and froglets. Alkaline phosphatase first appeared at the hatching tadpole stage. The liver showed the first band of alkaline phosphatase at st. III. This band migrated at 2.3∿2.4cm from the origin. A new band havin...

2006-01-01

 
 
 
 
61

Nucleotide and amino acid sequences of human intestinal alkaline phosphatase: close homology to placental alkaline phosphatase  

International Nuclear Information System (INIS)

A cDNA clone for human adult intestinal alkaline phosphatase (ALP) [orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1] was isolated from a ?gt11 expression library. The cDNA insert of this clone is 2513 base pairs in length and contains an open reading frame that encodes a 528-amino acid polypeptide. This deduced polypeptide contains the first 40 amino acids of human intestinal ALP, as determined by direct protein sequencing. Intestinal ALP shows 86.5% amino acid identity to placental (type 1) ALP and 56.6% amino acid identity to liver/bone/kidney ALP. In the 3'-untranslated regions, intestinal and placental ALP cDNAs are 73.5% identical (excluding gaps). The evolution of this multigene enzyme family is discussed

1987-01-01

62

Ultrastructural localisation of alkaline phosphatase in adult human large intestine.  

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The localisation of alkaline phosphatase in human large intestine was investigated at the ultrastructural level. Alkaline phosphatase was found in the mature absorptive cells of the surface, upper, and middle crypt epithelium; the reaction product was localised in the glycocalyx coat of the microvilli, the Golgi apparatus, and the rough endoplasmic reticulum. Slight activity was found in the immature absorptive cells of the middle and lower regions of the crypts. The undifferentiated cells an...

1982-01-01

63

Evolution of alkaline phosphatase in marine species of Vibrio.  

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The evolution of alkaline phosphatase was studied in marine species of Vibrio. Two antisera prepared against purified alkaline phosphatases from Vibrio splendidus and Vibrio harveyi were used to estimate the amino acid sequence divergence of this enzyme in 51 strains belonging to nine species. The methods used were the quantitative microcomplement fixation technique and the Ouchterlony double-diffusion procedure. There was a high degree of congruence between the measurement of the amino acid ...

Woolkalis, M. J.; Baumann, P.

1981-01-01

64

Studies on Alkaline Phosphatase Activity in Amphibians : III. Effects of Inhibitor and Temperature on Alkaline Phosphatases in Japanese Amphibians  

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The effects of L-homoarginine, L-phenylalanine, L-phenylalanylglycylglycine and L-leucine and high temperatures on the alkaline phosphatases extracted from the skins, lungs, livers, kidneys, ovaries and small intestines of Rana nigromaculata, Rana japonica, Rana rugosa, Rana limnocharis, Rana catesbeiana, Rhacophorus schlegelii, Hyla arborea japonica, Bufo japonicus and Cynops pyrrhogaster were examined in order to confirm the differences in the stability of alkaline phosphatases to these inh...

2006-01-01

65

Evaluation of Testosterone and Alkaline Phosphatase Activity Changes in Epidydimis of Toxoplasma gondii Infected Rats  

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Full Text Available Objective: Toxoplasma gondii is a widespread protozoan parasite that infects a broad range of warm blooded animals and humans. The present study was investigated to evaluate testosterone, alkaline phosphatase activity and malondialdehyde in male rats experimentally infected by Toxoplasma gondii, RH strain.Material & Methods: Male Wistar rats (n=20 were allocated into two groups, group one (n=10 that received 0.6 cc tachyzoites of T. gondii intraperitoneally (I.P., and control group (n=10 that received nothing. Animals were kept in standard conditions. 30 days after inducing Toxoplasma infection, 5cc blood was collected for assessment of serum testosterone, alkaline phosphatase and malondialdehyde levels. Epididymis tissues of Rats in whole groups were removed and prepared for analysis.Results: Alkaline phosphatase, and Testosterone were significantly increased in group that was infected by T.gondii in comparison to control group (P0.05.Epididymis weights in toxoplasmosis group was significantly decreased in comparison to control group (P<0.05. Positive brown alkaline phosphatase were observed in epididym tissue of infected toxoplasma group in comparison to control group.Conclusion: This study showed that T. gondii has augmenter effects on alkaline phosphatase activity, testosterone and has harmful effect on epididymis tissue.

Fatemeh Afshari

2013-07-01

66

Effects of orally administered cadmium on alkaline phosphatase isoenzymes in rat tissues.  

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Changes of alkaline phosphatase in small intestine, liver, kidney, bone and serum of rats administered 100 ppm (890 nM) of Cd2+ in the drinking water were observed during a 12 months period. After 2 weeks of cadmium administration, decreases in bone and small intestine alkaline phosphatases in serum of Cd-exposed rats were observed by a polyacrylamide gradient gel electrophoresis and the alterations continued through the 9th month of administration. At one month, the activities of the alkaline phosphatase fractions, p-1 and p-2, obtained from Sephadex G-200 column gel filtration of bone extracts from Cd-exposed rats were 32% and 43% of those in the controls, respectively, whereas Cd accumulation in the bone was very low (9 nmol/g wet weight). After 3 months, osteoporotic changes of bone and erosion of submucosa layer of the small intestine were observed by light microscopy. Alkaline phosphatase in small intestine of the Cd-exposed rats was 60% of that in the controls after 3 months. At 12 months, the decreased activity of bone alkaline phosphatase in Cd-exposed rats recovered to the same level as activity in the non-exposed rats. Moreover, the activity in kidney of the Cd-exposed rats was 80% of that in the controls. However, histological conversion from osteoporotic to osteomalacic changes in bone and kidney lesions by Cd administration were not observed by light microscopy. Liver alkaline phosphatase activity of the Cd-exposed rats did not change even at 12 months, whereas 1.2 mumol of Cd per g wet weight accumulated in this organ. PMID:4093840

Kobayashi, S; Kimura, M

1985-10-01

67

Influence of glucose fluctuations on alkaline phosphatase activity.  

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It has been suggested previously that non-enzymatic glycosylation of certain enzymes results in decreased enzymatic activity. In order to examine the role of high blood glucose concentrations (BG) on serum alkaline phosphatase (AP) activity, we determined BG and serum AP in 32 healthy children during an oral glucose tolerance test (OGTT) and in 10 type I diabetic children at 30-min intervals for at least 150 min. A significant negative correlation was noted for BG and AP during the oral glucose load (r = -0.57, p less than 0.01). In diabetics, however, only children with marked BG fluctuations showed this inverse relationship between BG and AP. These observations could be explained by the formation of a Schiff base as the initial step of non-enzymatic glycosylation of AP. This assumption was further confirmed by in vitro experiments, in which AP was incubated with glucose resulting in decreased enzymatic activity. The dialyzable aldimine formed initially is subsequently stabilized as ketoamine after long-term incubation. PMID:6475451

Pollak, A; Schober, E; Coradello, H; Lischka, A; Levin, S; Waldhauser, F; Lubec, G

1984-01-01

68

Sensitive optical detection of alkaline phosphatase activity with quantum dots  

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A simple method has been developed to detect the activity of alkaline phosphatase (ALP) by the changing of fluorescence intensities of the quantum dots (QDs). In this system, the fluorescence intensities of the QDs were quenched by p-nitrophenol (pNP) which was produced in the process of ALP catalytic reaction. A series of linear calibration curves of the activity of ALP were obtained in different pH buffer solutions. The wide linear range was 3–1000 U L{sup ?1} and the detection limit was 3 U L{sup ?1} (S/N=3). Furthermore, the experimental conditions of biosensor were optimized, and anti-interference ability was presented. The activity of ALP was also detected in serum and the recovery of ALP in serum samples was more than 95%. The excellent performance of this biosensor indicates that it can be used in practice detection of ALP. -- Highlights: • A sensitive ALP biosensor is constructed based on QDs without complex processes. • The analysis processing is very convenient, simple and rapid. • The detection mechanism of the ALP biosensor is studied by XPS. • The paper proposes a feasible approach for some substrates or enzymes detecting.

Ren, Xiangling [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China); The State Key Laboratory of Bioelectronics, Southeast University, Nanjing 210096 (China); Chen, Zhenzhen; Chen, Xiaoying; Liu, Jing [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China); Tang, Fangqiong, E-mail: tangfq@mail.ipc.ac.cn [Laboratory of Controllable Preparation and Application of Nanomaterials, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, No. 29, Zhongguancun East Road, Haidian District, Beijing 100190 (China)

2014-01-15

69

Sensitive optical detection of alkaline phosphatase activity with quantum dots  

International Nuclear Information System (INIS)

A simple method has been developed to detect the activity of alkaline phosphatase (ALP) by the changing of fluorescence intensities of the quantum dots (QDs). In this system, the fluorescence intensities of the QDs were quenched by p-nitrophenol (pNP) which was produced in the process of ALP catalytic reaction. A series of linear calibration curves of the activity of ALP were obtained in different pH buffer solutions. The wide linear range was 3–1000 U L?1 and the detection limit was 3 U L?1 (S/N=3). Furthermore, the experimental conditions of biosensor were optimized, and anti-interference ability was presented. The activity of ALP was also detected in serum and the recovery of ALP in serum samples was more than 95%. The excellent performance of this biosensor indicates that it can be used in practice detection of ALP. -- Highlights: • A sensitive ALP biosensor is constructed based on QDs without complex processes. • The analysis processing is very convenient, simple and rapid. • The detection mechanism of the ALP biosensor is studied by XPS. • The paper proposes a feasible approach for some substrates or enzymes detecting

2014-01-01

70

Isolation and properties of extracellular alkaline phosphatase from Bacillus intermedius.  

Science.gov (United States)

Alkaline phosphatase (APase) was isolated from the culture liquid of the streptomycin-resistant strain of Bacillus intermedius S3-19 and purified as a homogeneous preparation by ion-exchange chromatography and FPLC. Electrophoresis and gel-filtration revealed that the active enzyme is a monomer with molecular weight of 46-47 kD. The enzyme possessed phosphomonoesterase and phosphodiesterase activities with maximal levels at pH 9.5 and 55 degreesC and was stable until 60 degreesC at pH 8.0-10.0. The isolated APase exhibits a broad specificity towards a wide variety of substrates. The effect of divalent metal ions and other reagents on its catalytic activities was studied. It was concluded that alkaline phosphatase of B. intermedius is similar to the secreted alkaline phosphatases from other Bacillus species in its physicochemical and catalytic properties. PMID:9864452

Sharipova, M R; Balaban, N P; Mardanova, A M; Nekhotyaeva, N V; Dementyev, A A; Vershinina, O A; Garusov, A V; Leshchinskaya, I B

1998-10-01

71

Identification of a butanol-extractable human placenta-specific antigen with alkaline phosphatase activity.  

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N-Butanol extracts of whole-term placenta from different individuals were prepared, and used as immunogens to raise heterologous hyperimmune sera in rabbits. Upon immunoelectrophoresis the anti-placenta antisera could recognize at least six antigenic components in the placental extract even after they had been completely absorbed with pooled male serum proteins. However, the antisera so absorbed, designated (-PMS) antisera, could still react strongly with several normal adult tissue extracts including kidney. Systematic and quantitative absorptions of the (-PMS) antisera were thus further carried out with individual butanol extracts of normal adult liver, lung, intestine, stomach, kidney, bone, pancreas, spleen, heart, cerebrum, cerebellum, breast, and packed red cells, as well as a composite extract containing equal amounts of each of the 13 adult tissue extracts. Of the six antigenic components in the placental extracts reacting with the (-PMS) antisera the only one which retained its reactivity with the antisera throughout exhaustive absorptions was associated with alkaline phosphatase activity. This immunologic and enzymologic identity was confirmed with homogeneous placental alkaline phosphatase. Extracts from each of three placentae injected into three pairs of rabbits all produced an identical antibody reaction with the unique determinant(s) of placental alkaline phosphatase. The same identity of precipitin reaction was also found with extracts of 14 other placentae against each of these antisera. It thus firmly establishes that placental alkaline phosphatase is a characteristic placenta-specific fetal protein. PMID:58937

Chang, C H; Angellis, D

1976-07-01

72

Evaluation of two new methods for routine measurement of alkaline phosphatase isoenzymes.  

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AIMS: To evaluate the performance of two new methods for the analysis of alkaline phosphatase (ALP) isoenzymes designed for use in the routine chemical pathology laboratory: pre-incubation with neuraminidase before agarose electrophoresis; and selective precipitation of the bone isoenzyme with wheat germ agglutinin (WGA). METHODS: Serum samples from 39 patients were analysed. Seventeen were from patients with liver disease, eight from patients with bone disease, and 14 from patients with norm...

1992-01-01

73

Osmoregulation of alkaline phosphatase synthesis in Escherichia coli K-12.  

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Alkaline phosphatase, the phoA product, is synthesized constitutively in phoR mutants. This constitutive synthesis, which is independent of phosphate control, varies with changes in the osmolarity of the growth medium; phoA expression increases with increasing osmolarity. Maximum expression of the osmoregulated genes phoA, ompC, and ompF was achieved by osmotic manipulation of minimal medium; complex media repressed their expression.

Villarejo, M.; Davis, J. L.; Granett, S.

1983-01-01

74

Asparagine-linked carbohydrate does not determine the cellular location of yeast vacuolar nonspecific alkaline phosphatase.  

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The nonspecific alkaline phosphatase of Saccharomyces sp. strain 1710 has been shown by phosphatase cytochemistry to be exclusively located in the vacuole, para-Nitrophenyl phosphate-specific alkaline phosphatase is not detected by this procedure because the activity of this enzyme is sensitive to the fixative agent, glutaraldehyde. To determine whether the oligosaccharide of nonspecific alkaline phosphatase is necessary to transport the enzyme into the vacuole, protoplasts were derepressed i...

Clark, D. W.; Tkacz, J. S.; Lampen, J. O.

1982-01-01

75

Trypanosoma rangeli: an alkaline ecto-phosphatase activity is involved with survival and growth of the parasite.  

Science.gov (United States)

The aim of this work was to investigate whether an alkaline ecto-phosphatase activity is present in the surface of Trypanosoma rangeli. Intact short epimastigote forms were assayed for ecto-phosphatase activity to study kinetics and modulators using ?-glycerophosphate (?-GP) and p-nitrophenyl phosphate (pNPP) as substrates. Its role in parasite development and differentiation was also studied. Competition assays using different proportions of ?-GP and pNPP evidenced the existence of independent and non-interacting alkaline and acid phosphatases. Hydrolysis of ?-GP increased progressively with pH, whereas the opposite was evident using pNPP. The alkaline enzyme was inhibited by levamisole in a non-competitive fashion. The Ca(2+) present in the reaction medium was enough for full activity. Pretreatment with PI-PLC decreased the alkaline but not the acid phosphatase evidence that the former is catalyzed by a GPI-anchored enzyme, with potential intracellular signaling ability. ?-GP supported the growth and differentiation of T. rangeli to the same extent as high orthophosphate (Pi). Levamisole at the IC50 spared significantly parasite growth when ?-GP was the sole source of Pi and stopped it in the absence of ?-GP, indicating that the alkaline enzyme can utilize phosphate monoesters present in serum. These results demonstrate the existence of an alkaline ecto-phosphatase in T. rangeli with selective requirements and sensitivity to inhibitors that participates in key metabolic processes in the parasite life cycle. PMID:23994113

Dos-Santos, André L A; Dick, Claudia F; Silveira, Thaís S; Fonseca-de-Souza, André L; Meyer-Fernandes, José R

2013-10-01

76

ALKALINE PHOSPHATASE ACTIVITY AS A MARKER OF DOG SEMEN FREEZABILITY  

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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.

KOSINIAK-KAMYSZ K.

2007-01-01

77

ALKALINE PHOSPHATASE ACTIVITY AS A MARKER OF DOG SEMEN FREEZABILITY  

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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.

K. KOSINIAK-KAMYSZ

2013-12-01

78

Specific Immunoassays for Placental Alkaline Phosphatase As a Tumor Marker  

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Human placental (hPLAP) and germ cell (PLAP-like) alkaline phosphatases are polymorphic and heat-stable enzymes. This study was designed to develop specific immunoassays for quantifying hPLAP and PLAP-like enzyme activity (EA) in sera of cancer patients, pregnant women, or smokers. Polyclonal sheep anti-hPLAP antibodies were purified by affinity chromatography with whole hPLAP protein (ICA-PLAP assay) or a synthetic peptide (aa 57–71) of hPLAP (ICA-PEP assay); the worki...

Stinghen, Se?rvio T.; Moura, Juliana F.; Zancanella, Patri?cia; Rodrigues, Giovanna A.; Pianovski, Mara A.; Lalli, Enzo; Arnold, Dodie L.; Minozzo, Joa?o C.; Callefe, Luis G.; Ribeiro, Raul C.; Figueiredo, Bonald C.

2006-01-01

79

Effect of aluminum phosphate on alkaline phosphatase activity of polyurethane foam immobilized cyanobacteria.  

Science.gov (United States)

The impact of insoluble phosphorus such as aluminum and rock phosphate on alkaline phosphatase activity of polyurethane foam immobilized cyanobacteria was assessed. Polyurethane foam immobilized Nodularia recorded the highest alkaline phosphatase activity of 9.04 (m. mol p-nitrophenol released h(-1) mg(-1) protein) in vitro. A higher concentration of aluminum phosphate was recorded a 25% reduction in alkaline phosphatase activity, ammonia content, and available phosphorus in culture filtrate of polyurethane foam immobilized cyanobacteria. In general, immobilized cyanobacteria exhibited a higher alkaline phosphatase activity in rock phosphate than aluminum phosphate. PMID:16897258

Ramalingam, N; Prasanna, B Gowtham

2006-09-01

80

Flow injection system for potentiometric determination of alkaline phosphatase inhibitors  

International Nuclear Information System (INIS)

A simple flow injection system for potentiometric detection of alkaline phosphatase (ALP) activity has been developed and adapted for determination of selected inhibitors of this enzyme. In this system monofluorophosphate (MFP) has been applied as a specific ALP substrate. The use of this substrate enables application of fluoride ion selective electrode (FISE) as a detector of the product of the enzyme catalyzed reaction. Moreover, chemical stability and low cost of MFP enables the use of the substrate as a component of the carrier. This way, fluoride ions contained in this substrate define and stabilize baseline signal generated by the detector. Effects from several potential ALP inhibitors and interfering species were studied and discussed. The system allows inhibitive detection of beryllium and vanadate ions at ppb levels with relatively high selectivity, short time of analysis and high throughput of the system (near 8 samples h-1)

2006-09-01

 
 
 
 
81

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

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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 2weeks 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. PMID:24935183

Ball, Vincent

2014-09-01

82

Alkaline phosphatase activity of water column fractions and seagrass in a tropical carbonate estuary, Florida Bay  

Science.gov (United States)

Few phosphorus-depleted coastal ecosystems have been examined for their ability to hydrolyze phosphomonoesters. We examined seasonal (August 2006-April 2007) alkaline phosphatase activity in Florida Bay, a phosphorus-limited shallow estuary, using fluorescent substrate at low concentrations (?2.0 ?M). In situ dissolved inorganic and organic phosphorus levels and phosphomonoester concentrations were also determined. Water column alkaline phosphatase activity was partitioned into two particulate size fractions (>1.2 and 0.2-1.2 ?m) and freely dissolved enzymes (Thalassia testudinum. Our results indicate: (1) potential alkaline phosphatase activity in Florida Bay is high compared to other marine ecosystems, resulting in rapid phosphomonoester turnover times (˜2 h). (2) Water column alkaline phosphatase activity dominates, and is split equally between particulate and dissolved fractions. (3) Alkaline phosphatase activity was highest during cyanobacterial blooms, but not when normalized to chl a. These results suggest that dissolved, heterotrophic and autotrophic alkaline phosphatase activity is stimulated by phytoplankton blooms. (4) The dissolved alkaline phosphatase activity is relatively constant, while the particulate activity is seasonally and spatially dynamic, typically associated with phytoplankton blooms. (5) Phosphomonoester concentrations throughout the bay are low, even though potential hydrolysis rates are high. We propose that bioavailable dissolved organic P is hydrolyzed by dissolved and microbial alkaline phosphatase enzymes in Florida Bay. High alkaline phosphatase activity in the bay is also promoted by long hydraulic residence times. This background activity is primarily driven by carbon and phosphorus limitation of microorganisms, and regeneration of enzymes associated with cell lysis. Pulses of inorganic phosphorus and labile organic phosphorus and nitrogen may stimulate autotrophs, particularly cyanobacteria, which in turn promote biological activity that increase alkaline phosphatase activity of both autotrophs and heterotrophs in the bay.

Koch, Marguerite S.; Kletou, Demetris C.; Tursi, Rosanna

2009-08-01

83

Paget’s Disease Of The Spine With Low Bone Alkaline Phosphatase  

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Full Text Available We present a case of 35-year-old man who was referred to us with a preliminary diagnosis of multiple spinal metastasis. Laboratory studies have shown a high serum alkaline phosphatase and a low bone alkaline phosphatase levels. Spinal magnetic resonance scans demonstrated involvement of T9, T11 and L3 vertebral bodies. The involved vertebrae appeared hypointense on T1- and hyperintense on T2-weighted images and a radionuclide bone scan has shown increased uptake in involved vertebral bodies. A transpedicular open biopsy was considered necessary for accurate diagnosis. Histopathologic evaluation of the specimen revealed typical “mosaic pattern” of Paget’s disease. After surgery, urinary deoxypridinoline and pridinoline levels were tested and were higher than normal. The patient was given oral doses of alendronate, 40 mg per day and follow-up magnetic resonance scans at 6 months and 2 years demonstrated improvement in the signal intensity of the involved vertebral bodies. This case not only shows that Paget’s disease can occur in the setting of low bone AP but also shows that the clinical improvement can be monitored by improvement in magnetic resonance signal.

Ferda CAGAVI

2005-06-01

84

How Should an Increase in Alkaline Phosphatase Activity Be Interpreted?  

International Nuclear Information System (INIS)

Low-level laser therapy, commonly known as LLLT, is the application of low power, monochromatic, and coherent light to injuries and lesions to stimulate healing and give pain relief. There are conflicting reports in the literature regarding the role of ALP. Objective: this study aimed to compare the cellular responses of wounded human skin fibroblasts exposed to doses of 0.5 J/cm2, 2.5 J/cm2, 5 J/cm2, or 16 J/cm2 using LLLT with a Helium-Neon laser (632.8 nm, 18.8 mW power output, 2.07 mW/cm2 power density, and 3.4 cm diameter spot size or area 9.1?cm2) to elucidate the role of alkaline phosphatase (ALP) in cell proliferation. Methods: cellular responses to laser irradiation were evaluated using ALP enzyme activity, LDH membrane integrity, neutral red for cell proliferation, optical density at 540?nm, and basic fibroblast growth factor (bFGF) expression. Results: results suggest that an increase in ALP is negatively correlated with cell growth depending on the concentration of growth factors in the medium. Results also indicate that an increase in ALP may be related to cellular damage. Conclusion: since the exact role of ALP is unknown, the ALP enzyme activity assay should be considered in conjunction with other cell proliferation assays such as neutral red, optical density, or more specifically bFGF expression.

2007-01-01

85

Characterization of the phosphatidylinositol-glycan membrane anchor of human placental alkaline phosphatase  

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Placental alkaline phosphatase (orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1) is a member of a diverse group of membrane proteins whose attachment to the lipid bilayer is mediated by a phosphatidylinositol-glycan. To investigate structural aspects of the glycolipid anchor, cultured WISH cells were used because, they produce the enzyme in abundant quantities. When cell suspensions were incubated with purified phosphatidylinositol-specific phospholipase C, most of the placental alkaline phosphatase was released from membranes in a hydrophilic form. On incubation of the cells with (/sup 14/C)ethanolamine, (/sup 14/C)myristic acid, or myo(/sup 3/H)inositol, each was incorporated into the phosphatase near the carboxyl terminus, showing that these components, which are found in other phosphatidylinositol membrane-linked proteins, are also present in placental alkaline phosphatase.

Howard, A.D.; Berger, J.; Gerber, L.; Familletti, P.; Udenfriend, S.

1987-09-01

86

Characterization of the phosphatidylinositol-glycan membrane anchor of human placental alkaline phosphatase  

International Nuclear Information System (INIS)

Placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] is a member of a diverse group of membrane proteins whose attachment to the lipid bilayer is mediated by a phosphatidylinositol-glycan. To investigate structural aspects of the glycolipid anchor, cultured WISH cells were used because, they produce the enzyme in abundant quantities. When cell suspensions were incubated with purified phosphatidylinositol-specific phospholipase C, most of the placental alkaline phosphatase was released from membranes in a hydrophilic form. On incubation of the cells with [14C]ethanolamine, [14C]myristic acid, or myo[3H]inositol, each was incorporated into the phosphatase near the carboxyl terminus, showing that these components, which are found in other phosphatidylinositol membrane-linked proteins, are also present in placental alkaline phosphatase

1987-01-01

87

Effects of /sup 45/Ca on murine skeletal muscle. 2. Alterations in acid and alkaline phosphatases and glucose 6-phosphatase  

Energy Technology Data Exchange (ETDEWEB)

Swiss albino mice were injected intraperitoneally with 3.7x10/sup 4/ Bq and 7.4x10/sup 4/ Bq /sup 45/Ca/g body weight. /sup 45/Ca-treated mice were sacrificed on days 1, 3, 5, 7, 14 and 28 and activities of acid phosphatase, alkaline phosphatase and glucose 6-phosphatase bioassayed in diaphragm and gastrocnemius. Activities of acid and alkaline phosphatases decreased after the 1st day of /sup 45/Ca treatment in both the muscles compared with the normal controls. These two enzymes apparently do not contribute to myofiber necrosis in irradiated skeletal muscle. Glucose 6-phosphatase levels increased in the two irradiated muscles and with 7.4x10/sup 4/ Bq /sup 45/Ca dose as much as 20-fold and 7-fold elevations are recorded in diaphragm and gastrocnemius, respectively, indicating a radiation-induced stimulation of inhibition of glucose 6-phosphatase channelization for energy generation. The possible role of elevated glucose 6-phosphatase levels in glycogen accumulation on account of radiations in skeletal muscle has been discussed.

Asotra, K.; Katoch, S.S.; Krishan, K.; Malhotra, R.K. (Himachal Pradesh Univ., Simla (India). Dept. of Bio-sciences)

1983-01-01

88

Subcellular localization of alkaline phosphatase in Bacillus licheniformis 749/C by immunoelectron microscopy with colloidal gold.  

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Subcellular distribution of the alkaline phosphatase of Bacillus licheniformis 749/C was determined by an immunoelectron microscopy method. Anti-alkaline phosphatase antibody labeled with 15- to 18-nm colloidal gold particles (gold-immunoglobulin G [IgG] complex) were used for the study. Both the plasma membrane and cytoplasmic material were labeled with the gold-IgG particles. These particles formed clusters in association with the plasma membrane; in contrast, in the cytoplasm the particles...

Tinglu, G.; Ghosh, A.; Ghosh, B. K.

1984-01-01

89

The Detection of Alkaline Phosphatase Using an Electrochemical Biosensor in a Single-Step Approach  

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Full Text Available A one-step, single use, disposable Alkaline Phosphatase (ALP biosensor has been developed. It is based on the detection of phenol produced by an ALP enzymatic reaction. It can operate at 25 °C in a pH 10 medium. It measures ALP of 0–300 IU/L. The permissible concentrations of glucose, ascorbic acid and urea without interference are 10 mM/L, 5 mg/L and 400 mg/L, respectively. Experimental results are compared to those obtained by spectrophotometric measurements in bovine serum. Excellent linearity between the biosensor outputs and the ALP concentrations exists. The agreement between the measurements of this biosensor and the spectrophotometer is also outstanding.

Chung-Chiun Liu

2009-10-01

90

The influence of ionizing radiation at the alkaline phosphatase activity and calcium 45 transport under hypothyroid conditions  

International Nuclear Information System (INIS)

The experiments with 2-month-old hypothyroid male rats (mercasolil 10 mg/kg per os during 21 days) have shown that total gamma-irradiation ( 0,5 Gy) caused phased changes of alkaline phosphatase - its reduction in the first dates of the investigation (3, 10 and 30 days), normal activity in a 90 days and repeated drop of the activity at the end of the experiment (180 days). In a 3 and 10 days after irradiation it was revealed the diminish of the calcium-accumulated ability of the duodenum mucous membrane in the hypothyroid rats. So it was concluded that combined action of radiation and mercasolil leaded to a decrease in calcium 45 transport in the duodenum and reduction in the activity of thermo labile isoenzyme of alkaline phosphatase in blood serum

1997-01-01

91

Human prostatic acid phosphatase directly stimulates collagen synthesis and alkaline phosphatase content of isolated bone cells  

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Human prostatic acid phosphatase (hPAP) directly enhances the differentiated characteristics of isolated bone cells in vitro. This enzyme, when added to cell cultures for 24 h in vitro stimulates collagen synthesis and the production of alkaline phosphatase. The effects are dose dependent, with statistically significant effects occurring from 0.1-100 nM hPAP. Concentrations higher than 100 nM do not evoke greater effects. The maximal effect of hPAP occurs between 12 and 24 h of exposure. The cells stimulated to the greatest degree are osteoprogenitor cells and osteoblasts. Fibroblasts isolated from the same tissue show a lesser sensitivity to hPAP. hPAP has no detectable effect on cell proliferation, as measured by radiolabeled thymidine incorporation or total DNA synthesis. None of the observations reported in this work can be attributed to contaminating proteins in the hPAP preparation. hPAP was radiolabeled with 125I and was used for affinity binding and cross-linking studies. Scatchard analysis of specific binding indicated the presence of 1.0 X 10(5) high affinity binding sites/cell, with a Kd of 6.5 nM. Cross-linking studies demonstrated the presence of one 320-kDa binding complex. The pH profile and kinetic determinations of Km and maximum velocity for hPAP were similar to those previously reported, except for the finding of positive cooperativity of the substrate with the enzyme under the conditions of our assay. We believe that the direct stimulation of bone-forming cells by hPAP may contribute to the sclerotic nature of skeletal bone around sites of neoplastic prostatic metastases and that the effect of the enzyme is probably mediated by a plasma membrane receptor

1991-01-01

92

CAN PREOPERATIVE SERUM ALKALINE PHOPHATASE AND SERUM CALCIUM BE USED AS A PREDICTORS OF POSTOPERATIVE TETANY  

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Full Text Available A total of 100 cases hyperthyroid patients were selected for this prospective study. The hyperthyoidism was confirmed by thyroid function tests and scintigraphy. Of these100 patients 60(60% patients had elevated serum alkaline phosphatase(ALPand decreased bone mineral density (BMD and 20(20% patients had normal ALP and normal BMD. All 100 patients underwent totalthyroidectomy. 20 patients with normal ALP did not show a shift in calcium levels and 80 patientswith elevated ALP showed shift in calcium levels (1 to 1.5 gm% 40 (40% patients with their calcium levels less than 9.5gm% and elevated ALP developed carpopedal spasm in postoperativeperiod. Hence we conclude that the combination of low normal calcium (< 9.5% and elevated ALP has a high predictive value for postoperative pedagogical spasm .

Chandrasekaran Maharajan

2012-11-01

93

The fate of purified radio-labelled alkaline phosphatase from the liver in the organism  

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Alkaline phosphatase (AP) from dog liver was enriched by a factor of 5.444 in various steps. Rabbit antiserum to the purified AP was produced; 125-I was used then to radiolabel the highly purified AP. Four dogs were cholecystectomized and subsequently received an extracorporal drainage of the bile ducts. Decrease rate of total radio-activity and of PBI in the serum was determined in one dog; likewise in three other dogs before and one week after occlusion of their main bile ducts. In addition, radioactivity above the organs was measured in some animals at short intervals. In the dogs with main bile duct drainage, bile was collected continuously for up to 70 h, samples were taken, and residual bile plus native dog bile were re-infused into the distal choledochus catheter. Total radioactivity, PBI and immunoprecipitability with antibodies were determined in the bile and serum samples. AP, GOT, CPT and bilirubin were determined in some serum samples. In addition, total radioactivity excreted by urine was established. Results show injected 125-I-AP to be rapidly stored in the liver and not to be excreted via bile to a decisive extent. The fact that 125-I-AP is not excreted via bile is further indicated by the identical decrease rate of injected 125-I-AP in the serum in dogs with and without main bile duct occlusion. Injected 125-I-AP appears to be metabolized very rapidly in the liver as is indicated by the rapid decrease of immuno precipitability of 125-I-AP in the serum. (orig./MG)

1981-01-01

94

Alkaline phosphatase: a possible treatment for sepsis-associated acute kidney injury in critically ill patients.  

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Acute kidney injury (AKI) is a common disease in the intensive care unit and accounts for high morbidity and mortality. Sepsis, the predominant cause of AKI in this setting, involves a complex pathogenesis in which renal inflammation and hypoxia are believed to play an important role. A new therapy should be aimed at targeting both these processes, and the enzyme alkaline phosphatase, with its dual mode of action, might be a promising candidate. First, alkaline phosphatase is able to reduce inflammation through dephosphorylation and thereby detoxification of endotoxin (lipopolysaccharide), which is an important mediator of sepsis. Second, adenosine triphosphate, released during cellular stress caused by inflammation and hypoxia, has detrimental effects but can be converted by alkaline phosphatase into adenosine with anti-inflammatory and tissue-protective effects. These postulated beneficial effects of alkaline phosphatase have been confirmed in animal experiments and two phase 2a clinical trials showing that kidney function improved in critically ill patients with sepsis-associated AKI. Because renal inflammation and hypoxia also are observed commonly in AKI induced by other causes, it would be of interest to investigate the therapeutic effect of alkaline phosphatase in these nephropathies as well. PMID:24462020

Peters, Esther; Heemskerk, Suzanne; Masereeuw, Rosalinde; Pickkers, Peter

2014-06-01

95

Decrease in mucosal alkaline phosphatase: a potential marker of intestinal reperfusion injury.  

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Intestinal ischemia necessitates rapid re-establishment of blood flow to prevent irreversible anoxic tissue damage. However, reperfusion results in additional injury as a consequence of the generation of oxygen free radicals. To date, no clear-cut marker to differentiate between ischemia versus reperfusion injury is available. In this regard, previous studies from our laboratory utilizing a rat in vitro lipid peroxidation model demonstrated that the generation of free radicals resulted in the inactivation of only the intestinal brush border alkaline phosphatase enzyme, with no effect on other membrane-bound digestive enzymes. Current studies were designed to assess the possibility of alkaline phosphatase being a specific marker of the reperfusion injury in canine and human ex vivo ischemia/reperfusion models. Small bowels harvested from canines and organ donors were subjected to ischemia followed by reperfusion. Brush border membrane enzymes, alkaline phosphatase, sucrase, maltase, and gamma-glutamyl transpeptidase were assayed in mucosal extracts from intestines with ischemia versus reperfusion. In both experimental models, there was no change in any enzyme activity with warm ischemia alone. In contrast, alkaline phosphatase activity was significantly decreased in both the canine and human reperfusion models, with no change in specific activities of sucrase, maltase, and gamma-glutamyl transpeptidase. Our data indicate that the alkaline phosphatase enzyme activity may represent a potential marker of intestinal reperfusion injury and may permit quantitative assessments of therapeutic interventions in human intestinal reperfusion injury. PMID:10218763

Sisley, A C; Desai, T R; Hynes, K L; Gewertz, B L; Dudeja, P K

1999-04-01

96

Control of placental alkaline phosphatase gene expression in HeLa cells: induction of synthesis by prednisolone and sodium butyrate  

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HeLa S3 cells produce an alkaline phosphatase indistinguishable from the enzyme from human term placenta. The phosphatase activity in these cells was induced by both prednisolone and sodium butyrate. Both agents stimulated de novo synthesis of the enzyme. The increase in phosphatase activity paralleled the increase in immunoactivity and biosynthesis of placental alkaline phosphatase. The fully processed phosphatase monomer in control, prednisolone-treated or butyrate-treated cells was a 64.5 K polypeptide, measured by both incorporation of L-[35S]methionine into enzyme protein and active-site labeling. The 64.5K polypeptide was formed by the incorporation of additional N-acetylneuraminic acid moieties to a precursor polypeptide of 61.5K. However, this biosynthetic pathway was identified only in butyrate-treated cells. In prednisolone-treated cells, the processing of 61.5K to 64.5K monomer was accelerated, and the presence of the 61.5 precursor could only be detected by either neuraminidase or monensin treatment. Phosphatase mRNA which comigrated with the term placental alkaline phosphatase mRNA of 2.7 kilobases was induced in the presence of either prednisolone or butyrate. Alkaline phosphatase mRNA is untreated HeLa S3 cells migrated slightly faster than the term placental alkaline phosphatase mRNA. Butyrate also induced a second still faster migrating alkaline phosphatase mRNA. Both prednisolone and butyrate increased the steady-state levels of placental alkaline phosphatase mRNA. The data indicate that the increase in phosphatase mRNA by prednisolone and butyrate resulted in the induction of alkaline phosphatase activity and biosynthesis in HeLa S3 cells. Furthermore, both agents induced the expression of different alkaline phosphatase gene transcripts without altering its protein product

1987-06-16

97

CASE REPORT: ALKALINE PHOSPHATASE, ?-GLUTAMYLTRANSFERASE, UREA AND CREATININE SERUM CONCENTRATION IN RABBITS (Oryctolagus cuniculus CONCENTRAÇÃO SÉRICA DE FOSFATASE ALCALINA, GAMA-GLUTAMIL TRANSFERASE, URÉIA E CREATININA EM COELHOS (Oryctolagus cuniculus  

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The enzymes ?-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 diagnóstico das hepatopatias. A uréia e a creatinina são excretadas através da urina. O aumento dos valores destes metabólitos em nível sérico é subsídio para diagnóstico de alteração da função renal em mamíferos. Procurou-se estabelecer, no presente estudo, valores de referência destas enzimas para coelhos (Oryctolagus cuniculus, visando subsidiar dados para o uso desses animais de laboratório em experimentos científicos. Para tanto, foi realizada colheita sangüínea de 45 animais, três amostras por coelho, totalizando 135 amostras, obtendo-se o soro por centrifugação imediata. Realizou-se o método colorimétrico e obtiveram-se valores de 36,44±10,66mg/dl de uréia, com mínimo de 9,24mg/dl e máximo de 66,06mg/dl; 0,94±0,22mg/dl para a creatinina, com mínimo de 0,51mg/dl e máximo de 1,53mg/dl e 72,41±29,68UI para fosfatase alcalina, com mínimo de 10,66UI e máximo de 167,39UI. A gama-glutamil transferase foi determinada pelo método cinético, revelando o valor de 6,85±3,31UI, mínimo de 2,0UI e máximo de 15,0UI.

PALAVRAS-CHAVES: Bioquímica sérica, coelhos, função hepática, função renal.

Mauren Picada Emanuelli

2008-04-01

98

Alkaline phosphatase and risk of stroke among Japanese: the Circulatory Risk in Communities Study (CIRCS).  

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Although serum alkaline phosphatase (ALP) levels have been associated with mortality from all-cause and from either ischemic or hemorrhagic stroke, no study has been published of the associations between ALP and the incidence of stroke. We therefore examined the associations of ALP with risk of stroke among Japanese, stratified by drinking status because ALP is known as an enzyme affected by alcohol consumption. We conducted a prospective cohort study of 10,754 Japanese subjects (4098 men and 6656 women) aged 40-69 years and living in 4 communities under systematic surveillance for stroke incidence. During the 16-year follow-up, we documented 264 strokes (164 ischemic strokes and 69 hemorrhagic strokes) for men and 225 strokes (118 ischemic strokes and 89 hemorrhagic strokes) for women. There was a U-shaped association between ALP level and stroke incidence in both men and women, which was confined primarily to nondrinkers. For nondrinkers, higher ALP levels were associated with an elevated risk of ischemic stroke for men and of hemorrhagic stroke for women, whereas lower ALP levels were associated with elevated risks of ischemic and hemorrhagic strokes in both men and women. Our data indicate that not only higher, but also lower, serum ALP level may be a predictor for the risk of stroke in nondrinking men and women. PMID:22841505

Shimizu, Yuji; Imano, Hironori; Ohira, Tetsuya; Kitamura, Akihiko; Kiyama, Masahiko; Okada, Takeo; Ishikawa, Yoshinori; Shimamoto, Takashi; Yamagishi, Kazumasa; Tanigawa, Takeshi; Iso, Hiroyasu

2013-10-01

99

Radioimmunoassay for prostatic acid phosphatase in human serum. Methodologic aspects  

International Nuclear Information System (INIS)

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

1982-01-01

100

Radioimmunoassay for prostatic acid phosphatase in human serum. Methodologic aspects  

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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.

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))

1982-01-01

 
 
 
 
101

Additional Possibility of Data Analysis of Enzyme Inhibition and Activation. 5. Comparative Study of Temperature Activation of Calf Alkaline Phosphatase and Escherichia coli Alkaline Phosphatase  

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Full Text Available It was shown that simultaneous account of a course of change in the maximum reaction rate (V and the Michaelis constant (Km by plotting their vector representations in the three-dimensional KmVt coordinate system allows additional analysis of the dynamics of enzyme temperature activation. It also makes it possible to study the mechanism of enzyme action under varying temperature conditions of technological processes by use of such new parameters of enzyme activation as: a enzyme activation intensity, b the overall enzyme activation effect, c a geometrical portrait of enzyme activation. A comparative study of temperature activation of calf alkaline phosphatase and Escherichia coli alkaline phosphatase was performed by conventional and new methods of data processing.

V.I. Krupyanko

2005-01-01

102

Differentiating Intracellular from Extracellular Alkaline Phosphatase Activity in Soil by Sonication  

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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....

2013-01-01

103

Construction of an alkaline phosphatase-liposome system: a tool for biomineralization study.  

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Alkaline phosphatase is required for the mineralization of bone and cartilage. This enzyme is localized in the matrix vesicle, which plays a role key in calcifying cartilage. In this paper, we standardize a method for construction an alkaline phosphatase liposome system to mimic matrix vesicles and examine a some kinetic behavior of the incorporated enzyme. Polidocanol-solubilized alkaline phosphatase, free of detergent, was incorporated into liposomes constituted from dimyristoylphosphatidylcholine (DMPC), dilaurilphosphatidylcholine (DLPC) or dipalmitoylphosphatidylcholine (DPPC). This process was time-dependent and >95% of the enzyme was incorporated into the liposome after 4h of incubation at 25 degrees C. Although, incorporation was more rapid when vesicles constituted from DPPC were used, the incorporation was more efficient using vesicles constituted from DMPC. The 395nm diameter of the alkaline phosphatase-liposome system was relatively homogeneous and more stable when stored at 4 degrees C. Alkaline phosphatase was completely released from liposome system only using purified phosphatidylinositol-specific phospholipase C (PIPLC). These experiments confirm that the interaction between alkaline phosphatase and lipid bilayer of liposome is via GPI anchor of the enzyme, alone. An important point shown is that an enzyme bound to liposome does not lose the ability to hydrolyze ATP, pyrophosphate and p-nitrophenyl phosphate (PNPP), but a liposome environment affects its kinetic properties, specifically for pyrophosphate. The standardization of such system allows the study of the effect of phospholipids and the enzyme in in vitro and in vivo mineralization, since it reproduces many essential features of the matrix vesicle. PMID:12009304

Camolezi, Fernando L; Daghastanli, Katia R P; Magalhães, Prislaine P; Pizauro, João M; Ciancaglini, Pietro

2002-09-01

104

AVERRHOA CARAMBOLA (STAR FRUIT INDUCES HEPATIC ALKALINE PHOSPHATASE ACTIVITY IN RATS  

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Full Text Available The objective of the study is to examine the in vivo effect of star fruit juice at different storage conditions on the activity of alkaline phosphates in female Sprague Dawley (SD rats. A total of 20 healthy female rats weighing 180-200g were used for the experiment. All animals were divided into four groups with five animals per group (n=5. First control was served as control group. Second, third and forth groups were orally treated with a single dose daily of freshly prepared star fruit juice, juice stored for 1-hour and juice stored for 3-hour, respectively for 14 days. All animals were sacrificed at day-15 and various organs such as liver, kidney and heart were removed. All organs were homogenised and centrifuged to prepare cytosolic fraction as a source of alkaline phosphatase enzyme. Protein content of each organ was determined.  p-nitrophenol phosphate was used as a substrate to determine the activity of alkaline phosphatase using spectrophotometer. All results were analysed using Dunnett’s test. Based on the results obtained, a significant increase (P<0.01 of the activity of alkaline phosphatase in rat liver was observed in all star fruit juice treatment groups when compared to control rats. In conclusion, star fruit juice at different storage times was selectively induced the activity of alkaline phosphatase in rat liver but not in the heart and kidney.

Shamala F.

2011-01-01

105

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

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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.

Vijayaprasad K

2010-09-01

106

Effects of sodium nitroprusside activity of acid and alkaline invertases and alkaline phosphatase in lemongrass (Cymbopogon flexuosus Steud) Wats  

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Here we report the effects of SNP, a nitric oxide donor on sucrose metabolizing enzymes, acid and alkaline invertase (EC 3.2.1.26 and 3.2.1.153) and ubiquitous alkaline phosphatase (EC 3.1.3.1) in four lemongrass varieties viz., Krishna, Cauveri, Nima and Cheerharit. For the study, two 15 d lemongrass tillers were cut and immediately dipped into the test tubes containing SNP solution (5 mL) of variable strength (1 to 5 mM) and one without SNP (as control); kept for 4 h under mild sunlight. Th...

Deepak Ganjewala

2010-01-01

107

Electrophoresis of enzyme--monoclonal antibody complexes: studies of human placental alkaline phosphatase polymorphism.  

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Enzyme--monoclonal antibody complexes formed between six different monoclonal antibodies and the six phenotypes of human placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] that represent the homozygous and heterozygous combinations of the three common alleles have been examined by electrophoresis in starch, acrylamide, and agarose gels. Since the complexes formed retain full enzyme activity, they could be detected after gel electrophores...

Gogolin, K. J.; Slaughter, C. A.; Harris, H.

1981-01-01

108

High force eccentric exercise enhances serum tartrate-resistant acid phosphatase-5b and osteocalcin.  

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We investigated the effects of eccentric contractions (ECs) on bone metabolism markers and the relationship between bone metabolism and skeletal muscle related protein. Seventeen young untrained men were divided into two groups and performed either 60 or 30 maximal ECs. We measured serum levels of osteocalcin (OC), bone alkaline phosphatase, cross-linked N-telopeptide of type I collagen (NTx), and tartrate-resistant acid phosphatase 5b (TRACP-5b), growth hormone (GH), and insulin-like growth factor-1 (IGF-1). Blood samples were collected for up to five days after ECs. OC with 60 ECs were significantly higher than with 30 ECs (2 hours; pbone metabolism. Our results suggest that contraction-induced IGF-1 may activate OC and NTx in acute response. PMID:24583540

Tsuchiya, Y; Sakuraba, K; Ochi, E

2014-03-01

109

DNA polymorphism of alkaline phosphatase isozyme genes: Linkage disequilibria between placental and germ-cell alkaline phosphotase alleles  

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The use of human placental alkaline phosphatase (PLAP) cDNA as a probe allows the detection and identification of restriction DNA fragments derived from three homologous genes, i.e., intestinal alkaline phosphatase (AP), germ-cell AP (GCAP), and PLAP. In previous RFLP studies the authors have reported linkage disequilibria between an RsaI and two PstI (a and b) polymorphic restriction sites and electrophoretic types of PLAP. In this report they present evidence that, in spite of the strong correlation with PLAP types, PstI(b) is an RFLP of GCAP. The data indicate close linkage between the PLAP and GCAP loci. 18 refs., 2 figs., 3 tabs.

Beckman, G.; Beckman, L.; Sikstroem, C. (Univ. of Umea (Sweden)); Millan, J.L. (La Jolla Cancer Research Foundation, CA (United States))

1992-11-01

110

Cord blood calcium, phosphate, magnesium, and alkaline phosphatase gestational age-specific reference intervals for preterm infants  

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Full Text Available Abstract Background The objective was to determine the influence of gestational age, maternal, and neonatal variables on reference intervals for cord blood bone minerals (calcium, phosphate, magnesium and related laboratory tests (alkaline phosphatase, and albumin-adjusted calcium, and to develop gestational age specific reference intervals based on infants without influential pathological conditions. Methods Cross-sectional study. 702 babies were identified as candidates for this study in a regional referral neonatal unit. After exclusions (for anomalies, asphyxia, maternal magnesium sulfate administration, and death, relationships were examined between cord blood serum laboratory analytes (calcium, phosphate, magnesium, alkaline phosphatase, and albumin-adjusted calcium with gestation age and also with maternal and neonatal variables using multiple linear regression. Infants with influential pathological conditions were omitted from the development of gestational age specific reference intervals for the following categories: 23-27, 28-31, 32-34, 35-36 and > 36 weeks. Results Among the 506 preterm and 54 terms infants included in the sample. Phosphate, magnesium, and alkaline phosphatase in cord blood serum decreased with gestational age, calcium increased with gestational age. Those who were triplets, small for gestational age, and those whose mother had pregnancy-induced hypertension were influential for most of the analytes. The reference ranges for the preterm infants ? 36 weeks were: phosphate 1.5 to 2.6 mmol/L (4.5 to 8.0 mg/dL, calcium: 2.1 to 3.1 mmol/L (8.3 to 12.4 mg/dL; albumin-adjusted calcium: 2.3 to 3.2 mmol/L (9.1 to 12.9 mg/dL; magnesium 0.6 to 1.0 mmol/L (1.4 to 2.3 mg/dL, and alkaline phosphatase 60 to 301 units/L. Conclusions These data suggest that gestational age, as well as potentially pathogenic maternal and neonatal variables should be considered in the development of reference intervals for preterm infants.

Lyon Andrew W

2011-08-01

111

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

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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 this membrane passage in organ cultured intestinal mucosal explants. We found that in addition to a rapid uptake into the cytoplasm, a fraction of the fatty acid analogs were inserted directly into the brush border membrane. Furthermore, a brief exposure of microvillar membrane vesicles to a fat mixture mimicking a physiological solution of dietary mixed micelles, rearranged the lipid raft microdomain organization of the membranes. Thus, the fat mixture generated a low-density subpopulation of microvillar detergent resistant membranes (DRMs) highly enriched in alkaline phosphatase (AP). Since this GPI-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 microdomains of the brush border.

Hansen, Gert H; Rasmussen, Karina

2011-01-01

112

Activity of Follicular Fluid Phosphatases and Their Correlation with Levels of Serum Esteroidal Hormones and Gonadotropins  

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Full Text Available Introduction: The aim of this study was to evaluate the correlation between serum levels of ovarian and gonadotropin hormones, age and number of follicles with follicular alkaline and acid phosphatase activity in infertile women under controlled ovarian hyperstimulation. Methods: After collection of follicular fluid and calculation of the number of follicles, the specific activity of alkaline (ALP and acid phosphatase (ACP was determined according to the total protein in 19 women at the time of puncture. Also at that time, the levels of progesterone, estradiol, and follicle stimulating hormone (FSH and leuteinizing hormone (LH of their sera were measured. The correlation of follicular ALP and ACP with each serum hormone levels, women age and number of follicles was calculated using non-parametric analysis. Results: The ALP has a correlation with progesterone (P=0.01 levels but doesn’t have any correlation with the other factors. However, the ACP activity has a correlation not only with follicular number but also with estradiol and progesterone levels (P=0.05. Conclusion: Thus ACP activity is more affected by ovarian hormone than ALP and it can affect the ovarian microenvironment and oocyte development.

Sh Byranvand

2006-10-01

113

Membrane-bound alkaline phosphatase from ectopic mineralization and rat bone marrow cell culture.  

Science.gov (United States)

Cells from rat bone marrow exhibit the proliferation-differentiation sequence of osteoblasts, form mineralized extracellular matrix in vitro and release alkaline phosphatase into the medium. Membrane-bound alkaline phosphatase was obtained by method that is easy to reproduce, simpler and fast when compared with the method used to obtain the enzyme from rat osseous plate. The membrane-bound alkaline phosphatase from cultures of rat bone marrow cells has a MW(r) of about 120 kDa and specific PNPP activity of 1200 U/mg. The ecto-enzyme is anchored to the plasma membrane by the GPI anchor and can be released by PIPLC (selective treatment) or polidocanol (0.2 mg/mL protein and 1% (w/v) detergent). The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10. This fraction hydrolyzes ATP (240 U/mg), ADP (350 U/mg), glucose 1-phosphate (1100 U/mg), glucose 6-phosphate (340 U/mg), fructose 6-phosphate (460 U/mg), pyrophosphate (330 U/mg) and beta-glycerophosphate (600 U/mg). Cooperative effects were observed for the hydrolysis of PPi and beta-glycerophosphate. PNPPase activity was inhibited by 0.1 mM vanadate (46%), 0.1 mM ZnCl2 (68%), 1 mM levamisole (66%), 1 mM arsenate (44%), 10 mM phosphate (21%) and 1 mM theophylline (72%). We report the biochemical characterization of membrane-bound alkaline phosphatase obtained from rat bone marrow cells cultures, using a method that is simple, rapid and easy to reproduce. Its properties are compared with those of rat osseous plate enzyme and revealed that the alkaline phosphatase obtained has some kinetics and structural behaviors with higher levels of enzymatic activity, facilitating the comprehension of the mineralization process and its function. PMID:16798036

Simão, Ana Maria S; Beloti, Márcio M; Cezarino, Rodrigo M; Rosa, Adalberto Luiz; Pizauro, João M; Ciancaglini, Pietro

2007-04-01

114

Anopheles gambiae alkaline phosphatase is a functional receptor of Bacillus thuringiensis jegathesan Cry11Ba toxin.  

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Alkaline phosphatases (ALPs, EC 3.1.3.1) isolated from lepidopteran and dipteran species are identified as receptors for Cry1Ac and Cry11Aa toxins, respectively [Jurat-Fuentes, J. L., and Adang, M. J. (2004) Eur. J. Biochem. 7, 3127-3135; Fernandez, L. E., et al. (2006) Biochem. J. 396, 77-84]. In our study, an alkaline phosphatase cDNA (AgALP1) was cloned from the midgut of Anopheles gambiae larvae. The encoded 63 kDa protein has a predicted glycosylphosphatidylinositol (GPI) anchor omega-site ((526)Asp), an N-glycosylation site ((239)Asn-Leu-Thr), and an O-glycosylation site ((312)Ser). AgALP1(t) was expressed in Escherichia coli and used to prepare antiserum and to analyze the interaction of AgALP with mosquitocidal Cry11Ba toxin. Anti-AgALP serum localized AgALP to the apical brush border in the anterior and posterior midgut of larvae and detected a 65 kDa species on a blot of brush border membrane vesicles (BBMVs) protein prepared from larvae. ALP activity was released from larval BBMVs prepared by phosphatidylinositol-specific phospholipase C (PIPLC) treatment, and after separation by two-dimensional gel electrophoresis and blotting, a chain of doublet spots at 65 kDa was detected by anti-AgALP. A subset of these doublet spots bound Cry11Ba on a reprobed blot. Heterologously expressed AgALP1(t) bound [(125)I]Cry11Ba on dot blots and reduced the level of binding of [(125)I]Cry11Ba to brush border membrane vesicles by 41%, a percentage comparable to that of unlabeled Cry11Ba and aminopeptidase AgAPN2(t1) peptide. AgALP1(t) binds Cry11Ba toxin with a high affinity (23.9 nM) and shares a binding site on Cry11Ba with AgAPN2(t1). In bioassays against An. gambiae larvae, the presence of AgALP1(t) reduced larval mortality from 78 to 8%. We conclude that AgALP1 is a binding protein and a functional receptor for Cry11Ba toxin. PMID:19747003

Hua, Gang; Zhang, Rui; Bayyareddy, Krishnareddy; Adang, Michael J

2009-10-20

115

Effects of sodium nitroprusside activity of acid and alkaline invertases and alkaline phosphatase in lemongrass (Cymbopogon flexuosus Steud Wats  

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Full Text Available Here we report the effects of SNP, a nitric oxide donor on sucrose metabolizing enzymes, acid and alkaline invertase (EC 3.2.1.26 and 3.2.1.153 and ubiquitous alkaline phosphatase (EC 3.1.3.1 in four lemongrass varieties viz., Krishna, Cauveri, Nima and Cheerharit. For the study, two 15 d lemongrass tillers were cut and immediately dipped into the test tubes containing SNP solution (5 mL of variable strength (1 to 5 mM and one without SNP (as control; kept for 4 h under mild sunlight. The results revealed that moderate SNP concentration (2 mM was most effective, caused drastic reduction (40% in protein content in var. Nima followed by Krishna (33%, Cauveri (17% and Cheerharit (12%. In contrast, SNP (1 mM has impressively enhanced protein content in all the lemongrass varieties. The SNP (2 mM markedly inhibited the activity of acid invertase by 38% in Cheerharit, 35% Nima and 28% Cauveri whereas and alkaline invertase by 21, 28 and 24% respectively in var. Cheerharit, Nima and Krishna. Similarly, SNP (5 mM severely inhibited (~ 63% the activity of the ALP in lemongrass var. Cauveri and Nima, 50% in Krishna and relatively less 23% in Cheerharit as compared to the control. However, in var. Nima, 50% loss in ALP activity had already been occurred after 2 mM SNP treatment. These results primarily suggests that NO interferes sucrose metabolism by anonymously hindering the activity of acid and alkaline invertase and ubiquitous alkaline phosphatase in lemongrasses.

Deepak Ganjewala

2010-01-01

116

Método fosfatasa alcalina anti-fosfatasa alcalina para el diagnóstico de inmunodeficiencias celulares Alkaline phosphatase-anti-alkaline phosphatase method for the diagnosis of cell immunodeficiencies  

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Full Text Available Para el estudio de 25 pacientes con infecciones recurrentes y un grupo control de 25 individuos supuestamente sanos, se aplicó, en nuestro laboratorio, el método inmunocitoquímico de fosfatasa alcalina anti-fosfatasa alcalina, para la cuantificación de las principales subpoblaciones de linfocitos T identificados con los anticuerpos monoclonales: anti-CD3, anti-CD4 y anti-CD8. Se obtuvieron diferencias significativas para las subpoblaciones TCD3 y CD4 positivos (p For the study of 25 patients with recurrent infections and a control group of 25 supposedly healthy individuals, our Laboratory applied the immunocytochemical method of alkaline phosphatase - anti-alkaline phosphatase for the quantitation of the main T-lymphocyte subgroups identified with monoclonal antibodies:antiCD3, anti-CD4 and anti-CD8. There were significant differences in positive TCD3 and CD4 subsets (p<0,05. Because this is a low cost and quick method, it may be applied by other immunodiagnosis labs throughout the country

Berta B Socarrás Ferrer

2001-12-01

117

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

Energy Technology Data Exchange (ETDEWEB)

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.

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

1998-01-01

118

Rescue of Severe Infantile Hypophosphatasia Mice by AAV-Mediated Sustained Expression of Soluble Alkaline Phosphatase  

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Hypophosphatasia (HPP) is an inherited disease caused by a deficiency of tissue-nonspecific alkaline phosphatase (TNALP). The major symptom of human HPP is hypomineralization, rickets, or osteomalacia, although the clinical severity is highly variable. The phenotypes of TNALP knockout (Akp2-/-) mice mimic those of the severe infantile form of HPP. Akp2-/- mice appear normal at birth, but they develop growth failure, epileptic seizures, and hypomineralization and die by 20 days of age. Previou...

Matsumoto, Tae; Miyake, Koichi; Yamamoto, Seiko; Orimo, Hideo; Miyake, Noriko; Odagaki, Yuko; Adachi, Kumi; Iijima, Osamu; Narisawa, Sonoko; Milla?n, Jose? Luis; Fukunaga, Yoshitaka; Shimada, Takashi

2011-01-01

119

Bovine Intestinal Alkaline Phosphatase Attenuates the Inflammatory Response in Secondary Peritonitis in Mice  

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Lipopolysaccharide (LPS) contributes importantly to morbidity and mortality in sepsis. Bovine intestinal alkaline phosphatase (BIAP) was demonstrated to detoxify LPS through dephosphorylation. LPS injection combined with BIAP reduced inflammation and improved survival in various experimental settings. In this study, single-dose intravenous administration of BIAP (0.15 IU/g) was applied in a murine cecal ligation and puncture (CLP) model of polymicrobial sepsis. Saline was given as control (S ...

Veen, Suzanne Q.; Vliet, Arle?ne K.; Wulferink, Marty; Brands, Ruud; Boermeester, Marja A.; Gulik, Thomas M.

2005-01-01

120

Pseudohypophosphatasia: aberrant localization and substrate specificity of alkaline phosphatase in cultured skin fibroblasts.  

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We explored the biochemical basis for the disorder pseudohypophosphatasia (PsHYPT) in one patient by examining the substrate specificity and localization of alkaline phosphatase (ALP) in cultured dermal fibroblasts. Despite substantial ALP activity, in cell homogenates, toward the artificial substrate 4-methyl-umbelliferyl phosphate (4-MUP), there was a marked deficiency in ALP activity toward the natural substrates pyridoxal 5'-phosphate (PLP) and phosphoethanolamine (PEA), indicating altere...

Fedde, K. N.; Cole, D. E.; Whyte, M. P.

1990-01-01

 
 
 
 
121

Lipid accumulation and alkaline phosphatase activity in human preadipocytes isolated from different body fat depots  

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BACKGROUND: Alkaline phosphatase (ALP) controls intracellular lipid accumulation in human preadipocytes, but it is not known whether ALP is expressed in all body fat depots, or whether it has a similar role at all sites. DESIGN: Cross-sectional. SETTING AND SUBJECTS: Subjects undergoing breast reduction and abdominal fat biopsies operations at Charlotte Maxeke Johannesburg Academic Hospital. OUTCOME MEASURES: This study compared intracellular lipid accumulation and ALP activity in th...

Ali, A. T.; Ferris, W. F.; Penny, C. B.; Merwe, M. T.; Jacobson, B. F.; Paiker, J. E.; Crowther, N. J.

2013-01-01

122

Intestinal alkaline phosphatase is a gut mucosal defense factor maintained by enteral nutrition  

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Under conditions of starvation and disease, the gut barrier becomes impaired, and trophic feeding to prevent gut mucosal atrophy has become a standard treatment of critically ill patients. However, the mechanisms responsible for the beneficial effects of enteral nutrition have remained a mystery. Using in vitro and in vivo models, we demonstrate that the brush–border enzyme, intestinal alkaline phosphatase (IAP), has the ability to detoxify lipopolysaccharide and prevent bacterial invasion ...

Goldberg, Ross F.; Austen, William G.; Zhang, Xiaobo; Munene, Gitonga; Mostafa, Golam; Biswas, Shaluk; Mccormack, Michael; Eberlin, Kyle R.; Nguyen, John T.; Tatlidede, Hamit S.; Warren, H. Shaw; Narisawa, Sonoko; Milla?n, Jose L.; Hodin, Richard A.

2008-01-01

123

Plasma alkaline phosphatase as a sensitive indicator of age and skeletal development in wild coscoroba swans  

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Context. Recent studies have suggested that plasma alkaline phosphatase (ALP) can be used to assess skeletal development as well as health status in wild animals. Aims. However, the information about age-related dynamics of ALP in birds, especially in precocial species, is very scarce. Therefore, before ALP measurements can be effectively interpreted, it is necessary to determine its normal variation for each species, age group and sex. Methods. Here, we report total-ALP levels of free-living...

2010-01-01

124

Intestinal origin alkaline phosphatase activity in plasma for differential diagnosis of jaundice.  

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Intestinal origin alkaline phosphatase activity (ALP) in plasma was measured by a sensitive immunocapture assay in 104 jaundiced patients--84 with intrahepatic and 20 with post-hepatic jaundice. Increased enzyme activities were observed in those with intrahepatic disease and subnormal values in those with post-hepatic disease. At a discriminant level intestinal origin ALP showed a diagnostic sensitivity of 77% for intrahepatic cholestasis, with a diagnostic accuracy of 75% for its differentia...

1991-01-01

125

In vitro osteogenesis assays: influence of the primary cell source on alkaline phosphatase activity and mineralization.  

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In trabecular bone fracture repair in vivo, osteogenesis occurs through endochondral ossification under hypoxic conditions, or through woven bone deposition in the vicinity of blood vessels. In vitro osteogenesis assays are routinely used to test osteoblastic responses to drugs, hormones, and biomaterials for bone and cartilage repair applications. These cell culture models recapitulate events that occur in woven bone synthesis, and are carried out using primary osteoblasts, osteoblast precursors such as bone marrow-derived mesenchymal stromal cells (BMSCs), or various osteoblast cell lines. With time in culture, cell differentiation is typically assessed by examining levels of alkaline phosphatase activity (an early osteoblast marker) and by evaluating the assembly of a collagen (type I)-containing fibrillar extracellular matrix that mineralizes. In this review, we have made a comparative analysis of published osteogenic assays using calvarial cells, calvaria-derived cell lines, and bone marrow stromal cells. In all of these cell types, alkaline phosphatase activity shows similar progression over time using a variety of osteogenic and mineralizing media conditions; however, levels of alkaline phosphatase activity are not proportional to observed mineralization levels. PMID:18842361

Hoemann, C D; El-Gabalawy, H; McKee, M D

2009-06-01

126

Use of site-directed mutagenesis to elucidate the role of arginine-166 in the catalytic mechanism of alkaline phosphatase.  

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The guanidinium group of arginine-166 has been postulated to act as an electrophilic species during phosphorylation of alkaline phosphatase. Its role could be either to stabilize the developing negative charge on the oxygen of the leaving group or the pentacoordinate transition state or to help bind the -PO2-3 group. We have produced via site-directed mutagenesis two Escherichia coli alkaline phosphatase mutants (lysine-166 and glutamine-166) to test whether the guanidinium group plays a crit...

Butler-ransohoff, J. E.; Kendall, D. A.; Kaiser, E. T.

1988-01-01

127

Effect of MPG on the radiation induced changes in the intestinal activity of alkaline phosphatase in mice  

International Nuclear Information System (INIS)

Adult male Swiss mice were exposed to 250, 500 and 1000 R of gamma rays with or without a prior intraperitoneal injection of 20 mg/kg MPG. Alkaline phosphatase activity in the ileum was estimated at different post irradiation intervals. The changes in the enzyme activity was dose dependent. MPG reduced the radiation induced increase in the alkaline phosphatase activity of the intestine. (author)

1981-04-01

128

Structural studies of human alkaline phosphatase in complex with strontium: Implication for its secondary effect in bones  

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Strontium is used in the treatment of osteoporosis as a ranelate compound, and in the treatment of painful scattered bone metastases as isotope. At very high doses and in certain conditions, it can lead to osteomalacia characterized by impairment of bone mineralization. The osteomalacia symptoms resemble those of hypophosphatasia, a rare inherited disorder associated with mutations in the gene encoding for tissue-nonspecific alkaline phosphatase (TNAP). Human alkaline phosphatases have four m...

2006-01-01

129

The Study Of Serum Prostate Specific Antigen And Phosphatase Isoenzymes Activity As Diagnostic Parameters In Patients With Prostate Cancer In Nigeria  

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Full Text Available Serum activities of Acid Phosphatase (ACP and Prostatic Acid Phosphatase (PAP are still employed in most hospitals in Nigeria for the diagnosis of prostate cancer, because of lack of resources for prostate specific antigen (PSA assay. Serum PSA and activities of phosphatase isoenzymes ACP and PAP, Alkaline Phosphatase (ALP and Heat stable Alkaline Phosphatase (HSAP were studied in 71 apparently healthy male controls and 47 proven prostate cancer patients. There were statistically significant increases in the mean serum levels of PSA, PAP, ACP, ALP and HSAP in the prostate cancer patients compared to the controls (P<0.001. PSA level was increased above the cut-off level in 85.1% of patients, PAP in 66.0%, ACP in 57.5%, ALP in 34.0% and HSAP in 21.3% of cases. Serum levels of PSA, ACP and PAP were lower and of ALP and HSAP higher in patients with longer duration of the disease (P<0.05. The study confirms the relevance of PSA assay over ACP, PAP, ALP and HSAP in the diagnosis of prostate cancer patients. It highlights the need for the inclusion of PSA assay in hospitals for accurate diagnosis of prostatic carcinoma.

Igwe CU

2004-10-01

130

Ozone inhalation in rats: effects on alkaline phosphatase and lactic dehydrogenase isoenzymes in lavage and plasma  

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Ozone is found in urban and rural atmospheres and is produced from a variety of natural and man-made sources. Animal studies conducted at typical ambient levels result in reproducible morphological, biochemical and functional effects. Ozone damages type I epithelial cells, induces proliferation of type II cells and produces inflammation of the terminal bronchiolar-alveolar duct region. Ozone increases lung oxygen utilization and increases glutathione metabolism. Ozone increases airway resistance. The authors measured lactic dehydrogenase (LD) isoenzymes to ascertain the tissue giving rise to the increased LD activity in lavage. They also assayed acid phosphatase, alkaline phosphatase, creatine kinase activities, and protein levels since these parameters were increased in rat lung lavage after particulate exposure. They determined white cell differential and red cell morphology parameters because previous investigators reported that ozone increased neutrophil/lymphocyte ratio.

Nachtman, J.P.; Moon, H.L.; Miles, R.C.

1988-10-01

131

An Alkaline Phosphatase Reporter Gene Assay for Induction of CYP3A4 In Vitro  

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Full Text Available CYP3A4 probably has the broadest catalytic activity of any cytochrome P450. It is a crucial task to test new drug candidates in a reliable system for their ability to induce expression of this enzyme. Firstly, a total of 300 bp core distal enhancer of CYP3A4 XREM region (-7972/-7673 were amplified from human genomic DNA. The PCR product was then ligated into a human secretory alkaline phosphatase cDNA-containing reporter vector (pSEAP2-1creating pX-SEAP2 plasmid. Secondly, 1143 bp of the CYP3A4 proximal promoter region (-1203/-61 was amplified from the genomic DNA and then ligated into pX-SEAP2 plasmid DNA (between XREM and alkaline phosphatase gene, creating pXP-SEAP2 plasmid. Reporter constructs were then co-transfected with an hPXR expression vector into human liver and intestinal cells in culture. Xenobiotic modulation of CYP3A4 promoter activity was determined by chemiluminescent secretory alkaline phosphatase assay. Significant CYP3A4 induction atthe transcriptional level using three different cell lines and four classical CYP3A4 inducers was observed. Transfection of reporter constructs in HepG2, HuH7 and Caco-2 cells, in general, produced similar pattern of induction by the same drugs with the exception ofphenobarbital. The results suggest that, carefully designed reporter gene systems can provide a useful in vitro approach for characterization of possible CYP3A4 inducers.

Hossein Hamzeiy

2006-01-01

132

Phytoplankton and bacterial alkaline phosphatase activity in the northern Adriatic Sea  

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The importance of bacterial, phytoplankton and dissolved alkaline phosphatase activity (APA) in the northern Adriatic was investigated during 2006. In surface waters total APA increased from early spring (0.07–0.08 µmol/l/h) to late spring (up to 4.64 µmol/l/h) and remained relatively high during the summer (0.46–0.71 µmol/l/h1), due to an increase in specific phytoplankton (up to 30 nmol/µg C/h) and bacterial APA (up to 17.11 nmol/µg C/h). Activity of free enzymes was not important....

2010-01-01

133

Biocompatibility and Alkaline Phosphatase Activity of Phosphorylated Chitooligosaccharides on the Osteosarcoma MG63 Cell Line  

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Phosphorylated chitooligosaccharides (P-COS) were prepared using a H3PO4, P2O5, Et3PO4 and hexanol solvent system. The P-COS were characterized by Fourier Transform Infrared Spectroscopy (FT-IR), Thermo gravimetric-Differential Thermal Analyzer (TG-DTA), 13C NMR, 31P NMR, X-ray diffraction analysis, solubility studies, biocompatibility and Alkaline Phosphatase Activity (ALP). The results reveal that phosphorylation occurred at the C3 and C6 position of OH groups and the C2 position of NH2 gro...

Jayachandran Venkatesan; Ratih Pangestuti; Zhong-Ji Qian; BoMi Ryu; Se-Kwon Kim

2010-01-01

134

Properties of a constitutive alkaline phosphatase from strain 74A of the mold Neurospora crassa  

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A constitutive alkaline phosphatase was purified to apparent homogeneity as determined by polyacrylamide gel electrophoresis from mycelia of the wild strain 74A of the mold Neurospora crassa, after growth on acetate and in the presence of saturating amounts of inorganic phosphate (Pi) for 72 h at 30ºC. The molecular mass was 58 kDa and 56 kDa as determined by exclusion chromatography and SDS-PAGE, respectively. This monomeric enzyme shows an apparent optimum pH ranging from 9.5 to 10.5 and M...

Morales, A. C.; Nozawa, S. R.; Thedei Jr, G.; Maccheroni Jr, W.; Rossi, A.

2000-01-01

135

Differences in sialic acid residues among bone alkaline phosphatase isoforms: a physical, biochemical, and immunological characterization.  

Science.gov (United States)

High-performance liquid chromatography (HPLC) separates three human bone alkaline phosphatase (BALP) isoforms in serum; two major BALP isoforms, B1 and B2, and a minor fraction, B/I, which is composed on average of 70% bone and 30% intestinal ALP. The current studies were intended to identify an in vitro source of the BALP isoforms for physical, biochemical, and immunological characterizations. The three BALP isoforms were identified in extracts of human osteosarcoma (SaOS-2) cells, by HPLC, after separation by anion-exchange chromatography. All three BALP isoforms were similar with respect to freeze-thaw stability, solubility, heat inactivation, and inhibition by L-phenylalanine, L-homoarginine, and levamisole. The isoforms were also kinetically similar (i.e., maximal velocity and KM at pH 8.8 and pH 10.0). The isoforms differed, however, with respect to sensitivity to precipitation with wheat germ agglutinin (WGA), P < 0.001, but not Concanavalin A. At 3.0 mg/ml, WGA precipitated approximately 25% of B/I but more than 80% of B1 and B2. Molecular weights were estimated by native gradient gel electrophoresis: B/I, 126 kDa; B1, 136 kDa; and B2, 141 kDa. Desialylation with neuraminidase reduced the apparent sizes of B1 and B2 to 127 kDa (i.e., approximately to that of B/I). The total carbohydrate content was calculated to be 18 kDa, 28 kDa, and 33 kDa (i.e., 14%, 21%, and 23%) for the BALP isofonns, B/I, B1, and B2, respectively. The number of sialic acid residues was estimated to be 29 and 45, for each B1 and B2 homodimer, respectively. Apparent discrepancies between these estimates of molecular weight and estimates based on gel filtration chromatography were attributed to nonspecific interactions between carbohydrate residues and the gel filtration beads. All three BALP isoforms showed similar dose-dependent linearity in the commercial Alkphase-B and Tandem-MP Ostase immunoassays, r = 0.944 and r = 0.985, respectively (P < 0.001). In summary, our data indicate that B1 and B2 have more (or more reactive) sialic acid residues compared with B/I, which mainly explains the apparent differences in molecular weight. Future investigations will focus on the clinical and functional significance of the revealed differences in sialic acid residues. PMID:12232676

Magnusson, P; Farley, J R

2002-12-01

136

Distribution of alkaline and acid phosphatases in the duodenal wall of native blackgoats by using different fixatives  

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Full Text Available Ten duodeni of adult goat were fixed in chilled acetone, 80% ethyl alcohol, alcohol-formalin solution, alcohol bouinssolution and buffered neutral formalin solution. The distribution of alkaline and acid phosphatases noticed in absorptive andgoblet cells that lining the duodenal mucosa of black goat, but different in their intensity and distribution according to differentfixatives. The distribution of alkaline phosphatase in absorptive columnar cells that lining intestinal glands was more intensethan other cells, whereas the concentration of acid phosphatase was more intense in goblet cells than other cells in the mucosaof goat duodenum specially in samples fixed in chilled acetone and ethyl alcohol 80%. The study revealed that the sampleswere fixed with chilled acetone gave highest reaction for alkaline and acid phosphatases than other fixative samples. Noreaction for alkaline and acid phosphatases included some absorptive cells lining villi, all cells lining the lower parts ofintestinal glands, paneth cells and submucosal glands in different fixatives, except submucosal glands revealed positivereaction for acid phosphatase in samples fixed in chilled acetone and 80% ethyl alcohol, paneth cells reveal positive reaction for the same enzyme in samples fixed in 80% ethyl alcohol in all examined areas of the duodenum wall of the native blackgoat.

N. S. Ahmed

2010-01-01

137

Increased ethanol accumulation from glucose via reduction of ATP level in a recombinant strain of Saccharomyces cerevisiae overexpressing alkaline phosphatase  

Science.gov (United States)

Background The production of ethyl alcohol by fermentation represents the largest scale application of Saccharomyces cerevisiae in industrial biotechnology. Increased worldwide demand for fuel bioethanol is anticipated over the next decade and will exceed 200 billion liters from further expansions. Our working hypothesis was that the drop in ATP level in S. cerevisiae cells during alcoholic fermentation should lead to an increase in ethanol production (yield and productivity) with a greater amount of the utilized glucose converted to ethanol. Our approach to achieve this goal is to decrease the intracellular ATP level via increasing the unspecific alkaline phosphatase activity. Results Intact and truncated versions of the S. cerevisiae PHO8 gene coding for vacuolar or cytosolic forms of alkaline phosphatase were fused with the alcohol dehydrogenase gene (ADH1) promoter. The constructed expression cassettes used for transformation vectors also contained the dominant selective marker kanMX4 and S. cerevisiae ?-sequence to facilitate multicopy integration to the genome. Laboratory and industrial ethanol producing strains BY4742 and AS400 overexpressing vacuolar form of alkaline phosphatase were characterized by a slightly lowered intracellular ATP level and biomass accumulation and by an increase in ethanol productivity (13% and 7%) when compared to the parental strains. The strains expressing truncated cytosolic form of alkaline phosphatase showed a prolonged lag-phase, reduced biomass accumulation and a strong defect in ethanol production. Conclusion Overexpression of vacuolar alkaline phosphatase leads to an increased ethanol yield in S. cerevisiae.

2014-01-01

138

Inhibitors of tissue-nonspecific alkaline phosphatase: design, synthesis, kinetics, biomineralization and cellular tests.  

Science.gov (United States)

Chronic kidney disease (CKD) is associated with numerous metabolic and endocrine disturbances, including abnormalities of calcium and phosphate metabolism and an inflammatory syndrome. The latter occurs early in the course of CKD and contributes to the development and progression of vascular calcification. A few therapeutic strategies are today contemplated to target vascular calcification in patients with CKD: vitamin K2, calcimimetics and phosphate binders. However, none has provided complete prevention of vascular calcification and there is an urgent need for alternate efficient treatments. Recent findings indicate that tissue-nonspecific alkaline phosphatase (TNAP) may represent a very promising drug target due to its participation in mineralization by vascular smooth muscle cells. We report the synthesis of four levamisole derivatives having better inhibition property on TNAP than levamisole. Their IC50, Ki and water solubility have been determined. We found that the four inhibitors bind to TNAP in an uncompetitive manner and are selective to TNAP. Indeed, they do not inhibit intestinal and placental alkaline phosphatases. Survival MTT tests on human MG-63 and Saos-2 osteoblast-like cells have been performed in the presence of inhibitors. All the inhibitors are not toxic at concentrations that block TNAP activity. Moreover, they are able to significantly reduce mineralization in MG63 and Saos-2 osteoblast-like cells, indicating that they are promising molecules to prevent vascular calcification. PMID:24183741

Debray, Julien; Chang, Lei; Marquès, Stéphanie; Pellet-Rostaing, Stéphane; Le Duy, Do; Mebarek, Saida; Buchet, René; Magne, David; Popowycz, Florence; Lemaire, Marc

2013-12-15

139

[Comparison of daily alkaline phosphatase activity of a cyanobacterium (Microcystis aeruginosa) and a diatom (Synedra capitata)].  

Science.gov (United States)

Alkaline phosphatase activity (ALP) (EC: 3.1.3.1) presents a nycthemeral variation in both Microcystis aeruginosa (cyanobacterium) and Synedra capitata (diatom) species. Nevertheless, a comparative study reveals differences between the enzymatic behaviour of these two species. ALP is 33 times higher in cyanobacteria than in diatoms under similar experimental conditions. Microcystis aeruginosa presents therefore a larger capacity for mineralizing organic phosphorus per unit of biomass. Under LD (16:8) conditions, diatoms show a higher enzymatic activity during the day time (around 0.12 mumol pNPP/mn/mg); on the contrary, cyanobacterial enzymatic activity is rather low during the day time and rises at the beginning of night time (around 3.5 mumol pNPP/mn/mg). Finally, the mean of ALP of Synedra capitata is maximal (around 0.12 mumol pNPP/mn/mg) under total darkness (DD) while the mean of enzymatic activity is maximal (around 3.58 mumol pNPP/mn/mg) under permanent light (LL) for the cyanobacteria. These observed differences in the alkaline phosphatase activity between Microcystis aeruginosa and Synedra capitata might, to some extent, explain the observed alternances within the planktonic settlements between algae and cyanobacteria in hypereutrophic lakes such as the Grangent reservoir (Loire). PMID:9247024

Giraudet, H; Berthon, J L; Buisson, B

1997-06-01

140

A missense mutation in the human liver/bone/kidney alkaline phosphatase gene causing a lethal form of hypophosphatasia  

International Nuclear Information System (INIS)

Hypophosphatasia is an inherited disorder characterized by defective bone mineralization and a deficiency of serum and tissue liver/bone/kidney alkaline phosphatase (L/B/K ALP) activity. Clinical severity is variable, ranging from death in utero (due to severe rickets) to pathologic fractures first presenting in adult life. Affected siblings, however, are phenotypically similar. Severe forms of the disease are inherited in an autosomal recessive fashion; heterozygotes often show reduced serum ALP activity. The specific gene defects in hypophosphatasia are unknown but are thought to occur either at the L/B/K ALP locus or within another gene that regulates L/B/K ALP expression. The authors used the polymerase chain reaction to examine L/B/K ALP cDNA from a patient with a perinatal (lethal) form of the disease. They observed a guanine-to-adenine transition in nucleotide 711 of the cDNA that converts alanine-162 of the mature enzyme to threonine. The affected individual, whose parents are second cousins, is homozygous for the mutant allele. Introduction of this mutation into an otherwise normal cDNA by site-directed mutagenesis abolished the expression of active enzyme, demonstrating that a defect in the L/B/K ALP gene results in hypophosphatasia and that the enzyme is, therefore, essential for normal skeletal mineralization

1988-01-01

 
 
 
 
141

Clinical Characteristics of Primary Epstein Barr Virus Hepatitis with Elevation of Alkaline Phosphatase and ?-Glutamyltransferase in Children  

Science.gov (United States)

Purpose The aim of the present study was to evaluate the clinical characteristics of the primary Epstein-Barr virus (EBV) hepatitis with elevation of both serum alkaline phosphatase (ALP) and ?-glutamyltransferase (?-GT) levels in children. Materials and Methods A retrospective study was performed by reviewing of the medical records of 36 patients who were diagnosed with primary EBV hepatitis. The patients were divided into 2 groups: patients with elevated serum ALP and ?-GT levels (group 1) and patients without (group 2). Results The classic features of infectious mononucleosis (fever, pharyngitis and/or tonsillitis, and cervical lymphadenitis) were seen in 20 (57.1%) of group 1 patients and 18 (50.0%) of group 2 patients. Hepatitis with elevated serum ALP and ?-GT levels were present in 14 (38.9%) of the all patients. Of these patients, Jaundice occurred in only 2 (5.6%). The mean levels of aspartate aminotransferase and alanine aminotransferase (ALT) as well as the number of patients with ALT greater than 400 IU/L were significantly different between the groups (177 IU/L vs. 94 IU/L, 418 IU/L vs. 115 IU/L, and 50.0% vs. 13.6%; p=0.001, p=0.001, p=0.026, respectively). The mean duration of elevated serum ALT levels was 17.5 days in group 1 and 9.0 days in group 2 (p=0.013). All patients recovered fully without any chronic or serious complications. Conclusion Primary EBV hepatitis with predominant biochemical abnormalities of the elevation of ALP and ?-GT is frequent and mostly anicteric. This may represent a benign disease, but a delay in recovery of liver function as well.

Yang, Soo In; Geong, Jwa Hye

2014-01-01

142

Serum prostate-specific acid phosphatase: development and validation of a specific radioimmunoassay  

International Nuclear Information System (INIS)

We describe radioimmunoassay for human prostatic acid phosphatase [orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2] 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. 125I-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 [125I]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 ?g/liter and for 12 patients with advanced prostatic carcinoma between 100 and 500 ?g/liter. The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group

1978-01-01

143

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

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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+}.

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

2012-07-01

144

Effect of Zinc from Zinc Sulfate on Ewes` Weight, Milk Yield, Zn Concentrations in Serum and Serum Alkaline Phosphates Activity of Varamini Ewes  

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Full Text Available This experiment was conducted to investigate the effect of feeding supplemental zinc (zinc sulfate in different levels (0, 15 and 30 mg/kg on ewes weight, milk production, Zn concentrations in serum and serum alkaline phosphates activity. Thirty lactating Varaminni ewes were assigned to three experimental groups according to their live body weights, milk production and lambs sex in a completely randomized design. Ewes were fed a basal diet containing alfalfa, wheat straw, cottonseed meal, barley grain, wheat bran, cracked corn and vitamin-mineral supplements at 3.2% of BW to meet NRC requirements for protein, energy, macro minerals and micro minerals. The basal diet contained 15 mg/kg Zn and Zinc sulfate was added to the basal diet to supply 30 or 45 mg/kg of dietary zinc. Milk yielded, milk composition and ewes` weight was recorded at 7 and 21 days intervals respectively. Samples of the blood were taken three times (0, 35 and 64 for determination of Concentration of Zn, Cu and Fe, Na, K, Ca in serum. Also serum alkaline phosphates concentration of ewes was measured. Milk yield, milk composition and ewes` weight of ewes were not affected by supplemental zinc (p>0.05. Alkaline phosphatase concentration was increased with supplemental zinc linearly and this increase was significant (p<0.05. Blood mineral concentration was not affected by treatment (p>0.05.

A. Zali

2008-01-01

145

A Disposable Alkaline Phosphatase-Based Biosensor for Vanadium Chronoamperometric Determination  

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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.

Ana Lorena Alvarado-Gámez

2014-02-01

146

Fluoride stimulates [3H]thymidine incorporation and alkaline phosphatase production by human osteoblasts  

International Nuclear Information System (INIS)

The effect of sodium fluoride on alkaline phosphatase (ALP) release and [3H]thymidine uptake by human osteoblasts in culture was investigated. Sodium fluoride stimulated both ALP release and [3H]thymidine uptake at concentrations of sodium fluoride greater than 250 mumol/L. This stimulation was similar in magnitude to that induced by 1,25-dihydroxycholecalciferol. The fluoride-induced increase in ALP was inhibited by verapamil, a calcium channel blocker. We conclude that sodium fluoride stimulates osteoblasts to proliferate and to release ALP. This stimulation by fluoride is dependent on calcium influx. Fluoride-induced stimulation of human osteoblasts may be relevant to its effect in enhancing bone formation in patients with osteoporosis

1990-01-01

147

Near-infrared fluorescence probe for the determination of alkaline phosphatase.  

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Water-soluble CuInS2 quantum dots (QDs) were directly synthesized in an aqueous solution with mercaptopropionic acid (MPA) as stabilizers, and were functionalized using tryptophan molecules to form tryptophan-functionalized CuInS2 QDs (W-CuInS2 QDs) that had a strong fluorescence emission around 689 nm. The fluorescence of W-CuInS2 QDs could be quenched by Cu(2+) and then the addition of pyrophosphate (PPi) could effectively turn on the quenched fluorescence due to the strong interaction between Cu(2+) and PPi. The alkaline phosphatase (ALP) could catalyze the hydrolysis of PPi that would disassemble the complex of PPi-Cu(2+)-PPi. Therefore, the recovered fluorescence could be quenched again by the addition of ALP. In this paper, we developed a novel near-infrared fluorescence probe for the simple and convenient assay of ALP. PMID:24388906

Liu, Siyu; Pang, Shu; Na, Weidan; Su, Xingguang

2014-05-15

148

A case of polycythemia with low neutrophilic alkaline phosphatase and chromosome abnormalities in atomic bomb survivor  

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A case of mild polycythemia with low neutophilic alkaline phosphatase in a short-distance group was reported. The patient was exposed 1.4 km from the center of explosion (estimated exposure dose, 330 rad). He suffered from acute symptoms such as vomiting, diarrhea, increase in temperature, loss of hair, poor appetite, and hemorrhage. In an examination of a-bomb survivors in 1969, his erythrocyte count was 622 x 104/mm3 and his hemoglobin level was 18.3 gm/dl. Later his erythrocyte count was sometimes over 550 x 104/mm3. Upon admission to a hospital for a detailed examination, a slight increase in erythrocyte count and hemoglobin level and low NAP values were observed. Bone marrow findings revealed a slight increase in erythroblasts. Chromosomal analysis of bone marrow cells and peripheral lymphocytes revealed various abnormalities, seemingly related to exposure to radiation. Low NAPS values continued for a long time, and the patient remained healthy. (Tsunoda, M.)

1978-01-01

149

Intestinal alkaline phosphatase is a gut mucosal defense factor maintained by enteral nutrition  

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Under conditions of starvation and disease, the gut barrier becomes impaired, and trophic feeding to prevent gut mucosal atrophy has become a standard treatment of critically ill patients. However, the mechanisms responsible for the beneficial effects of enteral nutrition have remained a mystery. Using in vitro and in vivo models, we demonstrate that the brush–border enzyme, intestinal alkaline phosphatase (IAP), has the ability to detoxify lipopolysaccharide and prevent bacterial invasion across the gut mucosal barrier. IAP expression and function are lost with starvation and maintained by enteral feeding. It is likely that the IAP silencing that occurs during starvation is a key component of the gut mucosal barrier dysfunction seen in critically ill patients.

Goldberg, Ross F.; Austen, William G.; Zhang, Xiaobo; Munene, Gitonga; Mostafa, Golam; Biswas, Shaluk; McCormack, Michael; Eberlin, Kyle R.; Nguyen, John T.; Tatlidede, Hamit S.; Warren, H. Shaw; Narisawa, Sonoko; Millan, Jose L.; Hodin, Richard A.

2008-01-01

150

A human monoclonal antibody specific to placental alkaline phosphatase, a marker of ovarian cancer.  

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Placental alkaline phosphatase (PLAP) is a promising ovarian cancer biomarker. Here, we describe the isolation, affinity-maturation and characterization of two fully human monoclonal antibodies (termed B10 and D9) able to bind to human PLAP with a dissociation constant (Kd) of 10 and 30 nM, respectively. The ability of B10 and D9 antibodies to recognize the native antigen was confirmed by Biacore analysis, FACS and immunofluorescence studies using ovarian cancer cell lines and freshly-frozen human tissues. A quantitative biodistribution study in nude mice revealed that the B10 antibody preferentially localizes to A431 tumors, following intravenous administration. Anti-PLAP antibodies may serve as a modular building blocks for the development of targeted therapeutic products, armed with cytotoxic drugs, radionuclides or cytokines as payloads. PMID:24247025

Ravenni, Niccolò; Weber, Marcel; Neri, Dario

2014-01-01

151

Reduced activity of alkaline phosphatase due to host-guest interactions with humic superstructures.  

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Nuclear Magnetic Resonance (NMR) spectroscopy was applied to directly study the interactions between the alkaline phosphatase enzyme (AP) and two different humic acids from a volcanic soil (HA-V) and a Lignite deposit (HA-L). Addition of humic matter to enzyme solutions caused signals broadening in (1)H-NMR spectra, and progressive decrease and increase of enzyme relaxation (T1 and T2) and correlation (?C) times, respectively. Spectroscopic changes were explained with formation of ever larger weakly-bound humic-enzyme complexes, whose translational and rotational motion was increasingly restricted. NMR diffusion experiments also showed that the AP diffusive properties were progressively reduced with formation of large humic-enzyme complexes. The more hydrophobic HA-L affected spectral changes more than the more hydrophilic HA-V. (1)H-NMR spectra also showed the effect of progressively greater humic-enzyme complexes on the hydrolysis of an enzyme substrate, the 4-nitrophenyl phosphate disodium salt hexahydrate (p-NPP). While AP catalysis concomitantly decreased NMR signals of p-NPP and increased those of nitrophenol, addition of humic matter progressively and significantly slowed down the rate of change for these signals. In agreement with the observed spectral changes, the AP catalytic activity was more largely inhibited by HA-L than by HA-V. Contrary to previous studies, in which humic-enzyme interactions were only indirectly assumed from changes in spectrophotometric behavior of enzyme substrates, the direct measurements of AP behavior by NMR spectroscopy indicated that humic materials formed weakly-bound host-guest complexes with alkaline phosphatase, and the enzyme catalytic activity was thereby significantly inhibited. These results suggest that the role of extracellular enzymes in soils may be considerably reduced when they come in contact with organic matter dissolved in the soil solution. PMID:23953249

Mazzei, Pierluigi; Oschkinat, Hartmut; Piccolo, Alessandro

2013-11-01

152

Identification and characterization of an extracellular alkaline phosphatase in the marine diatom Phaeodactylum tricornutum.  

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In phosphorus-deficient conditions, Phaeodactylum tricornutum releases an alkaline phosphatase (PtAPase) to the medium that is readily detectable by activity staining. Nucleic acid and amino acid sequence of this alkaline phosphatase (APase) was identified by performing proteomic analysis and database searches. Sequence alignment suggests that PtAPase belongs to the PhoA family, and it possesses key residues at the Escherichia coli PhoA active site. Quantitative PCR results indicate that the induction of APase mRNA transcription is very sensitive to phosphorus availability and population growth. The molecular mass of native PtAPase (148 kDa) determined by gel filtration chromatography indicates that PtAPase, like most PhoA, is homodimeric. Zn and Mg ions are essential cofactors for most PhoA enzymes; however, PtAPase activity did not require Zn ions. In fact, 5 mM Zn²?, Mo²?, Co²?, Cd²?, or Cu²? inhibited its enzymatic activity, whereas 5 mM Mn²?, Mg²?, or Ca²? enhanced its enzymatic activity. The responses of PtAPase to divalent metal ions were different from those of most PhoAs, but were similar to the PhoA in a marine bacterium, Cobetia marina. Phylogenetic analysis shows that homologs of PhoA are also present in other diatom species, and that they clustered in a unique branch away from other PhoA members. PtAPase may represent a novel class of PhoA that helps diatoms to survive in the ocean. Quantification of the PtAPase mRNA may help monitor the physiological condition of diatoms in natural environments and artificial bioreactors. PMID:23358911

Lin, Hung-Yun; Shih, Chi-Yu; Liu, Hung-Chun; Chang, Jeng; Chen, Ying-Lan; Chen, Yet-Ran; Lin, Han-Tso; Chang, Yu-Yung; Hsu, Chun-Hua; Lin, Han-Jia

2013-08-01

153

Robotic implementation of assays: tissue-nonspecific alkaline phosphatase (TNAP) case study.  

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Laboratory automation and robotics have "industrialized" the execution and completion of large-scale, enabling high-capacity and high-throughput (100 K-1 MM/day) screening (HTS) campaigns of large "libraries" of compounds (>200 K-2 MM) to complete in a few days or weeks. Critical to the success these HTS campaigns is the ability of a competent assay development team to convert a validated research-grade laboratory "benchtop" assay suitable for manual or semi-automated operations on a few hundreds of compounds into a robust miniaturized (384- or 1,536-well format), well-engineered, scalable, industrialized assay that can be seamlessly implemented on a fully automated, fully integrated robotic screening platform for cost-effective screening of hundreds of thousands of compounds. Here, we provide a review of the theoretical guiding principles and practical considerations necessary to reduce often complex research biology into a "lean manufacturing" engineering endeavor comprising adaption, automation, and implementation of HTS. Furthermore we provide a detailed example specifically for a cell-free in vitro biochemical, enzymatic phosphatase assay for tissue-nonspecific alkaline phosphatase that illustrates these principles and considerations. PMID:23860647

Chung, Thomas D Y

2013-01-01

154

Pst I restriction fragment length polymorphism of the human placental alkaline phosphatase gene in normal placentae and tumors  

International Nuclear Information System (INIS)

The structure of the human placental alkaline phosphatase gene from normal term placentae was studied by restriction enzyme digestion and Southern blot analysis using a cDNA probe to the gene for the placental enzyme. The DNA digests fall into three distinct patterns based on the presence and intensity of an extra 1.1-kilobase Pst I Band. The extra 1.1-kilobase band is present in 9 of 27 placenta samples, and in 1 of these samples the extra band is present at double intensity. No polymorphism was revealed by digestion with restriction enzymes EcoRI, Sma I, BamHI, or Sac I. The extra Pst I-digestion site may lie in a noncoding region of the gene because no correlation was observed between the restriction fragment length polymorphism and the common placental alkaline phosphatase alleles identified by starch gel electrophoresis. In addition, because placental alkaline phosphatase is frequently re-expressed in neoplasms, the authors examined tissue from ovarian, testicular, and endometrial tumors and from BeWo choriocarcinoma cells in culture. The Pst I-DNA digestion patterns from these cells and tissues were identical to those seen in the normal ovary and term placentae. The consistent reproducible digestion patterns seen in DNA from normal and tumor tissue indicate that a major gene rearrangement is not the basis for the ectopic expression of placental alkaline phosphatase in neoplasia

1987-01-01

155

Value of preoperative bone and liver scans and alkaline phosphatase in the evaluation of breast cancer patients.  

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We reviewed the results of 133 bone scans and 63 liver scans (computed tomography, liver-spleen radionuclide scan, or ultrasonography) obtained for the preoperative evaluation of breast cancer patients. Information on the preoperative staging of breast cancer (TNM classification) was available in 131 of 133 patients. Bone scans had a low preoperative yield as only 4 of 133 patients (3%) had a positive bone scan that correlated with the results of plain films. Only 1 of 63 patients had a liver scan suggestive of possible metastasis. We also found that the alkaline phosphatase level was not a good predictor of bone or liver metastases in breast cancer patients. In 103 patients with normal bone scans, the majority (54%) had elevated alkaline phosphatase levels; conversely, 9 of 30 patients (30%) with abnormal scans had normal alkaline phosphatase levels. Furthermore, 51 of 63 patients (81%) with elevated alkaline phosphatase levels had normal liver scans. Approximately $74,000 was spent on these liver and bone scans. PMID:8427400

Brar, H S; Sisley, J F; Johnson, R H

1993-02-01

156

[Leucine arylamidase, lactate dehydrogenase and alkaline phosphatase activity of the urine of normal subjects of infant age].  

Science.gov (United States)

Urinary activity of Leucine arylamidase, lactate dahydrogenase and Alkaline phosphatase in 14 healt subjects, ranging from 2 to 10 years are described. Some correlations between enzymatic activities, ratios enzymatic activities/creatininuria and enzymatic activities/dayly proteic clearance are investigated. PMID:7375016

Camerini, G; Castaldi, G; Menegatti, E

1980-04-01

157

Pst I restriction fragment length polymorphism of the human placental alkaline phosphatase gene in normal placentae and tumors  

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The structure of the human placental alkaline phosphatase gene from normal term placentae was studied by restriction enzyme digestion and Southern blot analysis using a cDNA probe to the gene for the placental enzyme. The DNA digests fall into three distinct patterns based on the presence and intensity of an extra 1.1-kilobase Pst I Band. The extra 1.1-kilobase band is present in 9 of 27 placenta samples, and in 1 of these samples the extra band is present at double intensity. No polymorphism was revealed by digestion with restriction enzymes EcoRI, Sma I, BamHI, or Sac I. The extra Pst I-digestion site may lie in a noncoding region of the gene because no correlation was observed between the restriction fragment length polymorphism and the common placental alkaline phosphatase alleles identified by starch gel electrophoresis. In addition, because placental alkaline phosphatase is frequently re-expressed in neoplasms, the authors examined tissue from ovarian, testicular, and endometrial tumors and from BeWo choriocarcinoma cells in culture. The Pst I-DNA digestion patterns from these cells and tissues were identical to those seen in the normal ovary and term placentae. The consistent reproducible digestion patterns seen in DNA from normal and tumor tissue indicate that a major gene rearrangement is not the basis for the ectopic expression of placental alkaline phosphatase in neoplasia.

Tsavaler, L.; Penhallow, R.C.; Kam, W.; Sussman, H.H.

1987-07-01

158

Leucine Aminopeptidase, ?-Glucosidase and Alkaline Phosphatase Activity Rates and Their Significance in Nutrient Cycles in Some Coastal Mediterranean Sites  

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In aquatic microbial ecology, knowledge of the processes involved in the turnover of organic matter is of utmost importance to understand ecosystem functioning. Microorganisms are major players in the cycling of nutrients (nitrogen, phosphorus) and carbon, thanks to their enzymatic activities (leucine aminopeptidase, LAP, alkaline phosphatase, AP, and ?-glucosidase, ?-GLU) on organic polymers (proteins, organic phosphates and polysaccharides, respectively). Estimates of the decomposition ra...

Gabriella Caruso

2010-01-01

159

Prognostic role of serum phosphatases in management of head-neck and cervix carcinoma  

International Nuclear Information System (INIS)

Serum enzymes assays have been performed for the diagnosis of cancer on the basis of particular organ involvement, stage of cancer and in following course of disease progression or regression by onco physician. Some serum enzymes like aspartate, alanine transaminase, CEA, ferritin T-glutamyl transaminase and alanine phosphatase after irradiation have been studied by various workers

2002-11-15

160

Biological Apatite Formed from Polyphosphate and Alkaline Phosphatase May Exchange Oxygen Isotopes from Water through Carbonate  

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The oxygen isotopic composition in bone mineral phosphate is known to reflect the local water composition, environmental humidity, and diet1. Once ingested, biochemical processes presumably equilibrate PO43- with "body water" by the many biochemical reactions involving PO43- 2. Blake et al. demonstrated that enzymatic release of PO43- from organophosphorus compounds, and microbial metabolism of dissolved orthophosphate, significantly exchange the oxygen in precipitated apatite within environmental water3,4, which otherwise does not exchange with water at low temperatures. One of the enzymes that can cleave phosphates from organic substrates is alkaline phosphastase5, the enzyme also associated with bone mineralization. The literature often states that the mineral in bone in hydroxylapatite, however the mineral in bone is carbonated apatite that also contains some fluoride6. Deprotonation of HPO32- occurs at pH 12, which is impossibly high for biological system, and the predominate carbonate species in solution at neutral pH is HCO3-. To produce an apatite mineral without a significant hydroxyl content, it is possible that apatite biomineralization occurs through a polyphosphate pathway, where the oxygen atom required to transform polyphosphate into individual phosphate ions is from carbonate: [PO3-]n + CO32- -> [PO3-]n-1 + PO43- + CO2. Alkaline phosphatase can depolymerise polyphosphate into orthophosphate5. If alkaline phosphatase cleaves an oxygen atom from a calcium-carbonate complex, then there is no requirement for removing a hydrogen atom from the HCO3- or HPO43- ions of body water to form bioapatite. A mix of 1 mL of 1 M calcium polyphosphate hydogel, or nano-particles of calcium polyphosphate, and amorphous calcium carbonate were reacted with alkaline phosphatase, and maintained at neutral to basic pH. After two weeks, carbonated apatite and other calcium phosphate minerals were identified by powder x-ray diffraction. Orthophosphate and unreacted polyphosphate species were detected by Raman and IR spectroscopy. The oxygen isotope data of the reactants and products will also be presented. The possibility that carbonate acts as an intermediate reagent, transferring the oxygen from water to phosphate in biological apatite mineral formation may explain why biological apatite exhibits a significant carbonate content, and how this mineral is formed with an insignificant hydroxyl content. 1 Kohn, M.J., and Cerling, T.E. Rev Mineral Geochem 2002 (48) 455 2 Kolodny, Y., Luz, B., Navon, O. Earth Planet Sci Lett 1983 (64) 398 3 Blake, R.E., O'Neil, J.R., Garcia, G.A. Geochim et Cosmochim Acta 1997 (61) 4411 4 Blake, R.E., Alt, J.C., and Martini, A.M. PNAS 2001 (98) 2148-2153 5 Liang, Y., and Blake, R.E. Geochim Cosmochim Acta 2009 (73) 3782) 6 Pasteris, J.D. et al. Biomaterials 2004 (35) 229 7 Omelon et al., PLoS ONE 2009 4(5), e5634

Omelon, S. J.; Stanley, S. Y.; Gorelikov, I.; Matsuura, N.

2011-12-01

 
 
 
 
161

Conjugated polyelectrolyte-based real-time fluorescence assay for alkaline phosphatase with pyrophosphate as substrate.  

Science.gov (United States)

The fluorescence of the anionic, carboxylate-substituted poly(phenylene ethynylene) polymer PPECO2 is quenched very efficiently via the addition of 1 equiv of Cu(2+). Addition of pyrophosphate (PPi) into the weakly fluorescent solution of PPECO2 and Cu(2+) induces recovery of the polymer's fluorescence; the recovery occurs because PPi complexes with Cu(2+), effectively sequestering the ion so it cannot bind to the carboxylate groups of the polymer. A calibration curve was developed that relates the extent of fluorescence recovery to [PPi], making the PPECO2-Cu(2+) system a sensitive and selective turn-on sensor for PPi. Using the PPECO2-Cu(2+) system as the signal transducer, a real-time fluorescence turn-off assay for the enzyme alkaline phosphatase (ALP) using PPi as the substrate is developed. The assay operates with [PPi] in the micromolar range, and it offers a straightforward and rapid detection of ALP activity with the enzyme present in the nanomolar concentration range, operating either in an end point or real-time format. Kinetic and product inhibition parameters are derived by converting time-dependent fluorescence intensity into PPi (substrate) concentration, thus allowing calculation of the initial reaction rates (v(o)). Weak, nonspecific fluorescence responses are observed concomitant to addition of other proteins to the assay solution; however, the signal response to ALP is demonstrated to arise from the ALP catalyzed hydrolysis of PPi to phosphate (Pi). PMID:18855416

Liu, Yan; Schanze, Kirk S

2008-11-15

162

Cloning and characterization of a cDNA coding for mouse placental alkaline phosphatase  

International Nuclear Information System (INIS)

Mouse alkaline phosphatase was partially purified from placenta. Data obtained by immunoblotting analysis suggested that the primary structure of this enzyme has a much greater homology to that of human and bovine liver ALPs than to the human placental isozyme. Therefore, a full-length cDNA encoding human liver-type ALP was used as a probe to isolate the mouse placental ALP cDNA. The cloned mouse cDNA is 2459 base pairs long and is composed of an open reading frame encoding a 524-amino acid polypeptide that contains a putative signal peptide of 17 amino acids. Homology at the amino acid level of the mouse placental ALP is 90% to the human liver isozyme but only 55% to the human placental counterpart. RNA blot hybridization results indicate that the mouse placental ALP is encoded by a gene identical to the gene expressed in mouse liver, kidney, and teratocarcinoma stem cells. This gene is therefore evolutionarily highly conserved in mouse and human

1987-01-01

163

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

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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.

I.R. Inyang

2011-05-01

164

Alkaline phosphatase-positive cells isolated from human hearts have mesenchymal stem cell characteristics  

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Full Text Available Tissue-specific resident cells have been identified as a promising population of progenitor cells for cell-based therapies. We describe here the isolation from adult human hearts of tissue nonspecific alkaline phosphatase-positive cells (ALPL+ cells with mesenchymal stem cell (MSC characteristics. Samples from 24 adult cadaveric donors were obtained from a valve bank. Mean total ischemia time was 21.5 ± 9.1 hours. The success rate for the isolation of human heart-derived cells by the explant culture technique was 70% for the right auricle (14 of 20 trials and 33% for the right ventricle (7 of 21 trials. The total auricle-derived cell population (TAD was used for the purification of ALPL+ cells. TAD and ALPL+ cells expressed markers for MSC and pericytes. TAD cells and ALPL+ cells differentiated into adipocytes, osteoblasts and chondroblasts, and ALPL+ cells expressed markers of these three lineages more strongly than TAD cells, as shown by RT-PCR. This population therefore has a greater potential for differentiation into mesechymal lineages than TAD cells. Both cell populations express some markers of cardiac progenitors, such as GATA4, CD117 and VEGF. ALPL+ cells expressed troponin T and ABCG2, which are also markers of the cardiac lineage. Heart samples from tissue banks could be considered as sources of MSC with putative commitment towards cardiac lineages, even after prolonged ischemia times.

Alessandra Melo de Aguiar

2011-10-01

165

Electrochemical immunosensor of tumor necrosis factor ? based on alkaline phosphatase functionalized nanospheres.  

Science.gov (United States)

A novel immunosensor for sensitive detection of tumor necrosis factor ? was reported. First of all, gold nanoparticles were uniformly assembled on the surface of poly (styrene-acrylic acid) nanospheres, which was used as the matrix to conjugate alkaline phosphatase (ALP). And then, the obtained composite was used as multi-enzyme functionalized label for immunoassay. Biocompatible polyaniline doped with poly (acrylic acid) was electro-polymerized at the glass carbon electrode to construct the matrix for the immobilization of antibody TNF-?. After the sandwich immunoreaction, the labeled ALP was used to hydrolyze ?-naphthyl phosphate to produce the electroactive ?-naphthol, which could be amperometrically detected. The results showed that the electrochemical signals were proportional to the logarithm of the antigen concentration in the range of 0.02-200.00 ng/mL with the detection limit of 0.01 ng/mL. The developed immunoassay showed high sensitivity, acceptable stability and reproducibility, which might have potentially broad applications in protein diagnostics and bioassay. PMID:20378330

Yin, Zhengzhi; Liu, Yan; Jiang, Li-Ping; Zhu, Jun-Jie

2011-01-15

166

Intestinal alkaline phosphatase promotes gut bacterial growth by reducing the concentration of luminal nucleotide triphosphates.  

Science.gov (United States)

The intestinal microbiota plays a pivotal role in maintaining human health and well-being. Previously, we have shown that mice deficient in the brush-border enzyme intestinal alkaline phosphatase (IAP) suffer from dysbiosis and that oral IAP supplementation normalizes the gut flora. Here we aimed to decipher the molecular mechanism by which IAP promotes bacterial growth. We used an isolated mouse intestinal loop model to directly examine the effect of exogenous IAP on the growth of specific intestinal bacterial species. We studied the effects of various IAP targets on the growth of stool aerobic and anaerobic bacteria as well as on a few specific gut organisms. We determined the effects of ATP and other nucleotides on bacterial growth. Furthermore, we examined the effects of IAP on reversing the inhibitory effects of nucleotides on bacterial growth. We have confirmed that local IAP bioactivity creates a luminal environment that promotes the growth of a wide range of commensal organisms. IAP promotes the growth of stool aerobic and anaerobic bacteria and appears to exert its growth promoting effects by inactivating (dephosphorylating) luminal ATP and other luminal nucleotide triphosphates. We observed that compared with wild-type mice, IAP-knockout mice have more ATP in their luminal contents, and exogenous IAP can reverse the ATP-mediated inhibition of bacterial growth in the isolated intestinal loop. In conclusion, IAP appears to promote the growth of intestinal commensal bacteria by inhibiting the concentration of luminal nucleotide triphosphates. PMID:24722905

Malo, Madhu S; Moaven, Omeed; Muhammad, Nur; Biswas, Brishti; Alam, Sayeda N; Economopoulos, Konstantinos P; Gul, Sarah Shireen; Hamarneh, Sulaiman R; Malo, Nondita S; Teshager, Abeba; Mohamed, Mussa M Rafat; Tao, Qingsong; Narisawa, Sonoko; Millán, José Luis; Hohmann, Elizabeth L; Warren, H Shaw; Robson, Simon C; Hodin, Richard A

2014-05-15

167

Effects of parathyroid hormone and calcitonin on alkaline phosphatase activity and matrix calcification in rabbit growth-plate chondrocyte cultures  

Energy Technology Data Exchange (ETDEWEB)

The effects of PTH and calcitonin (CT) on the expression of mineralization-related phenotypes by chondrocytes were examined. In cultures of pelleted growth-plate chondrocytes. PTH caused 60-90% decreases in alkaline phosphatase activity, the incorporation of {sup 45}Ca into insoluble material, and the calcium content during the post-mitotic stage. These effects of PTH were dose-dependent and reversible. In contrast, CT increased alkaline phosphatase activity, {sup 45}Ca incorporation into insoluble material, and the calcium content by 1.4- to 1.8-fold. These observations suggest that PTH directly inhibits the expression of the mineralization-related phenotypes by growth-plate chondrocytes, and that CT has the opposite effects.

Kato, Y.; Shimazu, A.; Nakashima, K.; Suzuki, F.; Jikko, A.; Iwamoto, M. (Osaka Univ. (Japan))

1990-07-01

168

PURIFICATION AND CHARACTERIZATION OF ALKALINE PHOSPHATASE FROM DOLICHOS LAB-LAB AND ITS INVITRO DEPHOSPHORYLATION ACTIVITY ON NUCLEIC ACIDS  

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Full Text Available Phosphatase serves several functions in plant metabolism including growth governance, phosphorous level control, starch breakdown etc. Alkaline phosphatases, acting at an alkaline pH 8, are a significant class of enzymes that catalyze release of phosphate esters especially. This enzyme study is so far limited only to animal source and partly to microbial sources, in terms of clinical research. Although it has been identified that plant as a source of this enzyme may be exploited, there always has been a challenge on the isolation and characterization of this enzyme and how pure it can be. This paper partly addresses the above problem, where the enzyme has been isolated from the seeds Dolichos lab-lab plant characterized and its purity was checked by HPLC. The purity obtained was 98% and the enzyme has been further analyzed for its activity on nucleic acids, which gave promising and positive results.

Praveen Kumar Vemuri et al

2012-09-01

169

An immunohistological study of testicular germ cell tumours using two different monoclonal antibodies against placental alkaline phosphatase.  

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Using two monoclonal antibodies directed against placental alkaline phosphatase (H17E2 and D20L) the immunohistological staining of testicular germ cell tumours was compared with that of a wide range of normal and malignant tissues. All seminomas and malignant teratomas tested gave strong positive labelling with H17E2 but were either negative or only patchily positive with D20L. Neither antibody gave any positive reaction on the normal tissues tested. All other malignancies were negative with...

Epenetos, A. A.; Travers, P.; Gatter, K. C.; Oliver, R. D.; Mason, D. Y.; Bodmer, W. F.

1984-01-01

170

Modulation of placental alkaline phosphatase activity and cytokeratins in human HN-1 cells by butyrate, retinoic acid, catecholamines and histamine.  

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The effects of butyrate and retinoic acid in combination with catecholamines or histamine on the HN-1 human head and neck squamous carcinoma cell line were investigated analysing cell proliferation, placental alkaline phosphatase (PLAP) activity, and relative cytokeratin content. Butyrate inhibited cell proliferation in agar, whereas retinoic acid induced a small inhibitory effect. Butyrate enhanced PLAP activity in a time related manner in contrast to retinoic acid, which had no significant ...

Bijman, J. T.; Wagener, D. J.; Rennes, H.; Wessels, J. M.; Ramaekers, F. C.; Den Broek, P.

1987-01-01

171

Cloning and Expression of the Alkaline Phosphatase Gene from the Persian Type Culture Collection Escherichia coli K-12  

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The structural gene for alkaline phosphatase (phoA) of E. coli K-12 strain obtained from the Persian type culture collection (PTCC 1268) was cloned into pTZ57R plasmid as cloning vector and pGEM-3Z plasmid as expression vector, respectively. The recombinant plasmids were confirmed by different restriction enzymes and determination of the nucleotide sequence. Protein expression was induced by isopropyl -D thiogalactopyranoside (IPTG) and was analyzed using polyacrylamide gel electrophoresis (P...

2005-01-01

172

Arginine Coordination in Enzymatic Phosphoryl Transfer: Evaluation of the Effect of Arg166 Mutations in Escherichia coli Alkaline Phosphatase†,‡  

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Arginine residues are commonly found in the active sites of enzymes catalyzing phosphoryl transfer reactions. Numerous site-directed mutagenesis experiments establish the importance of these residues for efficient catalysis, but their role in catalysis is not clear. To examine the role of arginine residues in the phosphoryl transfer reaction, we have measured the consequences of mutations to arginine 166 in Escherichia coli alkaline phosphatase on hydrolysis of ethyl phosphate, on individual ...

O Brien, Patrick J.; Lassila, Jonathan Kyle; Fenn, Timothy D.; Zalatan, Jesse G.; Herschlag, Daniel

2008-01-01

173

Effect of sex and age on the activities of lactate dehydrogenase and alkaline phosphatase in the lungs of rats.  

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Since toxicity studies among different laboratories generally involve rats of different sex and age, this study was conducted to investigate the effect of sex, age and animal to animal variation in the activities of lactate dehydrogenase and alkaline phosphatase from bronchoalveolar lavage fluid, bronchoalveolar cell lysate and lung homogenate. Correlation between numbers of bronchoalveolar cells recovered from lungs and enzyme activity in bronchoalveolar cell lysate or lung homogenate supern...

Lopez, A.; Yong, S.; Sharma, A.; Morwood-clark, M.; Lillie, L. E.; Albassam, M.

1986-01-01

174

Identification of a highly mobilizable subset of human neutrophil intracellular vesicles that contains tetranectin and latent alkaline phosphatase.  

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Tetranectin, a protein recently identified in a wide variety of human secretory cells (Christensen, L., and I. Clemmensen. 1989. Histochemistry. 92:29-35) was found to colocalize with latent alkaline phosphatase activity in fractions well separated from azurophil granules, specific granules, gelatinase-containing granules, and plasma membranes when postnuclear supernatants of nitrogen-cavitated neutrophils were fractionated on discontinuous Percoll density gradients. Stimulation of intact neu...

Borregaard, N.; Christensen, L.; Bejerrum, O. W.; Birgens, H. S.; Clemmensen, I.

1990-01-01

175

High-resolution analysis of Zn2+ coordination in the alkaline phosphatase superfamily by EXAFS and x-ray crystallography  

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Comparisons among evolutionarily related enzymes offer opportunities to reveal how structural differences produce different catalytic activities. Two structurally-related enzymes, E. coli alkaline phosphatase (AP) and X. axonopodis nucleotide pyrophosphatase/phosphodiesterase (NPP) have nearly identical binuclear Zn2+ catalytic centers, but show tremendous differential specificity for hydrolysis of phosphate monoesters or phosphate diesters. To determine if there are differences in Zn2+ coord...

Bobyr, Elena; Lassila, Jonathan K.; Wiersma-koch, Helen I.; Fenn, Timothy D.; Lee, Jason J.; Nikolic-hughes, Ivana; Hodgson, Keith O.; Rees, Douglas C.; Hedman, Britt; Herschlag, Daniel

2012-01-01

176

Localization of Alkaline Phosphatase and Cathepsin D during Cell Restoration after Colchicine Treatment in Primary Cultures of Fetal Rat Hepatocytes  

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Localization of alkaline phosphatase (ALP) and cathepsin D (CAPD) in primary cultures of fetal rat hepatocytes was examined using double immunofluorescent staining in order to investigate the relationship between lysosome movement and the fate of ALP during cell restoration after microtubule disruption by colchicine. At 3 hr and 24 hr after colchicine treatment, numerous coarse dots containing ALP were observed throughout the cytoplasm, and some of these showed colocalization with CAPD. At 48...

Chida, Kohsuke; Taguchi, Meiko

2011-01-01

177

Coupling technique of self-ordered ring and phosphorimetry for the determination of alkaline phosphatase and diseases prediction  

Science.gov (United States)

Rhodamine S could emit strong and stable room temperature phosphorescence (RTP) on polyamide membrane (PAM) in the presence of heavy atom perturber Pb2+. When Rhodamine S-piperidine solution was dropped on PAM, the red (Rhod.S)n-P-SOR (Rhod.S, (Rhod.S)n, P and SOR refer to alizarin red S, multiple Rhod.S molecules, piperidine and self-ordered ring, respectively) formed on PAM, leading to the enhancement of room temperature phosphorimetry (RTP) intensity (Ip, 117.2) of (Rhod.S)n-P-SOR system, which was 2.4 times higher than that without SOR (Ip, 48.1). Wheat germ agglutinin (WGA) was labelled with (Rhod.S)n-P-SOR by the -NH- of Rhod.S reacting with the -COOH of WGA to form WGA-(Rhod.S)n-P-SOR. The formation of WGA-AP-WGA-(Rhod.S)n-P-SOR in the affinity adsorption (AA) reaction carried out between the -COOH of WGA in WGA-(Rhod.S)n-P-SOR and the -NH2 of alkaline phosphatase (AP) caused the RTP intensity (?Ip) of the WGA-AP-WGA-(Rhod.S)n-P-SOR system 7.8 times larger than that without (Rhod.S)n-P-SOR. Therefore, the coupling technique of SOR and solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace AP has been established. This method possessed good selectivity, high sensitivity (Detection limit (L.D) was 3.4 × 10-16 g mL-1) and accuracy, and it has been applied to the determination of trace AP in human serum and the forecast of human diseases, and the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Besides, the mechanism of the coupling technique for the determination of AP was discussed.

Zhang, Li Hong; Zheng, Zhi Yong; Jiang, Shu-Lian; Cui, Ma-Lin; Jiao, Li; Lin, Xuan; Cai, Wen-Lian; Lin, Shao-Qin; liu, Jia-Ming

2012-11-01

178

Studies on alkaline and acid phosphatase activity of neutrophil leukicytes, 2  

International Nuclear Information System (INIS)

With a view to analyzing the inhibiting effect of anticancer drugs and irradiation on hematopoiesis in rabbits neutrophil (pseudoeosinophil) counts and the neutrophilic activities of alkaline phosphatase (AP) and acid phosphatase (SP) were serially followed up after drug administration or irradiation. The enzym activity was estimated histochemically, using azo-dye staining. Each rabbit was given cyclophosphamid (CP) (25mg/kg x 10, at intervals of 5 - 7 days ; 50mg/kg x 5, every day; or 100mg/kg x 1, i.m.), Thio-TEPA (4mg/kg x 1, i.m.), Vinblastin (VBT) (1mg/kg x 1, i.v.), 6MP (25mg/kg x 1, p.o.), or Mitomycin C (MMC) (1.5mg/kg x 1, i.v.). The results obtained were as follows : 1) The neutrophil counts became slightly elevated at 24 hrs, reached their nadir at 48 to 72 hrs, and recovered to normal in 5 to 6 days thereafter, except with 6 MP which produced no significant change but for a temporary elevation after dosages. 2) Except in the group administrated 6MP, which caused no significant hematorogical changes, the AP changes were similar in all of the animal groups : after temporary depression, it became elevated for 5 to 6 days, and recovered to normal about 9 days thereafter. 3) SP showed no changes in the 25mg/kg x 10 CP and the 6MP groups, it became elevated in 2 or 3 days after the administration of MMC, VBT, or Thio-TEPA to recover to normal in 5 to 10 days thereafter. 4) 60Co irradiation (1,000 rad/whole body x 1) led to a temporary ascent in phil count followed by a descent from the 6th day on, and then a slow recovery to normal. AP was elevated from the third to the sixth days, and, after a depression on the tenth day, it returned to normal 24 days after irradiation, while SP showed a continued elevation from the 2nd to the 13th day. (author)

1983-01-01

179

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

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Full Text Available 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 105 of domestic proveniance containing polychlorinated biphenyls (PCB for a period of 30 days. This preparation is equivalent to the foreign preparation Aroclor 1254. A dose of 100 µg/kg of Delor 105 was given to the animals of the experimental group. These animals were euthanised on day 17 postpartum (n=4 i. e. 5 days from the end of a 30-day period of application; on day 25 postpartum (n=5 i.e. 17 days from the last application of PCB; on day 34 postpartum (n=5, which was equivalent to day 28 from the last application. The ewes from the control group were euthanised on day 17 (n=3, day 25 (n=4 and on day 34 (n-4 postpartum. When evaluating alkaline phosphatase (ALP activity in the glandular cells of the endometrium in the control group, a statistically significant increase (P<0.01 was observed on day 25 and on day 34 (P<0.001 compared to day 17 postpartum. No statistically significant differences in alkaline phosphatase (ALP activity were observed (P>0.05 in the experimental group. The mean values of its activity in the observed period were below the level of values of day 17 in the control group. Acidic phosphatase activity in the glandular cells of the ewes' endometrium showed a statistically conclusive increase between day 17 and day 25 as well as day 34 postpartum (P<0.001. Acidic phosphatase density in the experimental group of ewes showed no statistically marked change (P>0.05 at the observed intervals postpartum. The discussion is focused on PCB effect on the activity of alkaline and acidic phosphatase in the glandular cells of the endometrium of ewes in the puerperal period.

Valocky I.

2005-01-01

180

Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation  

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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.

Ciancaglini P.

2006-01-01

 
 
 
 
181

Gingival crevicular fluid flow rate and alkaline phosphatase level as potential marker of active tooth movement.  

Science.gov (United States)

Background: Gingival Crevicular Fluid (GCF) changes occur during orthodontic tooth movement and this could serve as a potential indicator to the response to active treatment. Aim: The objective of the study is to assess the changes in the GCF volume and the levels of Alkaline Phosphatase (ALP) during early phase of tooth movement. Methods: 20 patients requiring all first premolar extractions were selected and treated with conventional straight wire mechanotherapy. Canine retraction was done using Nitinol closed coil springs. Maxillary canine on one side acted as experimental site while the contralateral canine acted as control. GCF was collected from around the canines before initiation of retraction, 1 hour after initiating canine retraction, 1 day, 7 days, 14 days and 21 days. GCF volume and the ALP levels were estimated and compared with the control side. Results: The results showed statistically significant changes in the GCF volume and ALP levels on the 7(th), 14(th) and 21(st) days at the experimental sides. The peak in the activity occurred on the 14(th) day of initiation of retraction. The GCF volume and ALP levels did not show any significant variations at the control sites where no retraction was done. Conclusions: It can be concluded that GCF volume and ALP levels may serve as an indicator to assess tooth movement dynamics in orthodontic therapy. Based on the available data and further studies, ALP levels in GCF may aid in developing a reliable non-invasive chair side test for assessing the prognosis and progress of orthodontic therapy. PMID:24984665

Alfaqeeh, S A; Sukumaran, Anil

2014-06-01

182

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Alkaline phosphatase (ALP) is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart h [...] omogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4). Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 ± 3.43 nmol p-nitrophenol·mg protein-1·min-1): mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern) by immunofluorescence. ALP was inhibited a) strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84%, respectively) and b) less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11%, respectively). ?-estradiol and caffeine (0.5 and 2 mM) had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36%, respectively). Propranolol (2 mM) tended to activate ALP activity and corticosterone activated (18%) and inhibited (13%) (0.5 and 2 mM, respectively). We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.

A., Mota; P., Silva; D., Neves; C., Lemos; C., Calhau; D., Torres; F., Martel; H., Fraga; L., Ribeiro; M.N.M.P., Alçada; M.J., Pinho; M.R., Negrão; R., Pedrosa; S., Guerreiro; J.T., Guimarães; I., Azevedo; M.J., Martins.

183

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity  

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Full Text Available Alkaline phosphatase (ALP is important in calcification and its expression seems to be associated with the inflammatory process. We investigated the in vitro acute effects of compounds used for the prevention or treatment of cardiovascular diseases on total ALP activity from male Wistar rat heart homogenate. ALP activity was determined by quantifying, at 410 nm, the p-nitrophenol released from p-nitrophenylphosphate (substrate in Tris buffer, pH 10.4. Using specific inhibitors of ALP activity and the reverse transcription-polymerase chain reaction, we showed that the rat heart had high ALP activity (31.73 ± 3.43 nmol p-nitrophenol·mg protein-1·min-1: mainly tissue-nonspecific ALP but also tissue-specific intestinal ALP type II. Both ALP isoenzymes presented myocardial localization (striated pattern by immunofluorescence. ALP was inhibited a strongly by 0.5 mM levamisole, 2 mM theophylline and 2 mM aspirin (91, 77 and 84%, respectively and b less strongly by 2 mM L-phenylalanine, 100 mL polyphenol-rich beverages and 0.5 mM progesterone (24, 21 to 29 and 11%, respectively. ?-estradiol and caffeine (0.5 and 2 mM had no effect; 0.5 mM simvastatin and 2 mM atenolol activated ALP (32 and 36%, respectively. Propranolol (2 mM tended to activate ALP activity and corticosterone activated (18% and inhibited (13% (0.5 and 2 mM, respectively. We report, for the first time, that the rat heart expresses intestinal ALP type II and has high total ALP activity. ALP activity was inhibited by compounds used in the prevention of cardiovascular pathology. ALP manipulation in vivo may constitute an additional target for intervention in cardiovascular diseases.

A. Mota

2008-07-01

184

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

Science.gov (United States)

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 ?-sheet core with 19 surrounding ?-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? 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 comprising 329 C atoms from completely buried residues, whereas that of VAP comprised 264 C atoms, which may maintain the stability of HaAP under low-salt conditions. These characteristics of HaAP may be responsible for its unique functional adaptation permitting activity over a wide range of salt concentrations.

Arai, Shigeki; Yonezawa, Yasushi; Ishibashi, Matsujiro; Matsumoto, Fumiko; Adachi, Motoyasu; Tamada, Taro; Tokunaga, Hiroko; Blaber, Michael; Tokunaga, Masao; Kuroki, Ryota

2014-01-01

185

Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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 i [...] nto 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.

Ciancaglini, P.; Simão, A.M.S.; Camolezi, F.L.; Millán, J.L.; Pizauro, J.M..

186

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

Science.gov (United States)

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 ?-sheet core with 19 surrounding ?-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(?) 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 comprising 329 C atoms from completely buried residues, whereas that of VAP comprised 264 C atoms, which may maintain the stability of HaAP under low-salt conditions. These characteristics of HaAP may be responsible for its unique functional adaptation permitting activity over a wide range of salt concentrations. PMID:24598750

Arai, Shigeki; Yonezawa, Yasushi; Ishibashi, Matsujiro; Matsumoto, Fumiko; Adachi, Motoyasu; Tamada, Taro; Tokunaga, Hiroko; Blaber, Michael; Tokunaga, Masao; Kuroki, Ryota

2014-03-01

187

Ontogeny and distribution of alkaline and acid phosphatases in the digestive system of California halibut larvae (Paralichthys californicus).  

Science.gov (United States)

Studies aimed to assess the digestive physiology of marine fish larvae under culture conditions are important to further understand the functional characteristics and digestive capacities of the developing larvae. Most studies to date concentrate on intestinal lumen digestion and little attention to the absorption process. Thus, the objectives of this study were to histochemically detect and quantify some of the enzymes responsible for absorption and intracellular digestion of nutrients in the anterior and posterior intestine of California halibut larvae. Alkaline and acid phosphatases were detected from the first days post-hatch (dph). Alkaline phosphatase maintained a high level of activity during the first 20 dph in both intestinal regions. Thereafter, a clear intestinal regionalization of the activity was observed with the highest levels occurring in the anterior intestine. Acid phosphatase activity gradually increased in both intestinal regions during development, and a regionalization of the activity was not observed until late in development, once the ocular migration began. Highest levels were observed in the anterior intestine at the end of metamorphosis concomitant with the stomach development. The results from this study show some morphological and physiological changes are occurring during larval development and a clear regionalization of the absorption process as the larvae develops. These ontological changes must be considered in the elaboration of diets according to the digestive capacity of the larvae. PMID:23508366

Zacarias-Soto, Magali; Barón-Sevilla, Benjamín; Lazo, Juan P

2013-10-01

188

A novel glycosylphosphatidylinositol-anchored alkaline phosphatase dwells in the hepatic duct of the pearl oyster, Pinctada fucata.  

Science.gov (United States)

Alkaline phosphatases are ubiquitous enzymes involved in many important biological processes. Mammalian tissue-nonspecific alkaline phosphatase (TNAP) has long been thought to play an important role in bone mineralization. In this study, we identified a full-length cDNA encoding a potential alkaline phosphatse from pearl oyster Pinctada fucata by RT-PCR and RACE and designated the encoded protein as PFAP. The sequence of PFAP shares an overall similarity of 67% with that of human TNAP. Prediction and analysis of its secondary and tertiary structure revealed that the PFAP contains two mammalian-specific regions, the crown domain, involved in collagen binding, and the calcium binding domain, which hint its potential ability to participate in biomineralization. RT-PCR and in situ hybridization showed that the PFAP mRNA distributes specifically in the hepatic duct of the digestive diverticula. These findings implied its possible role in calcium absorption and transportation. In vivo, PFAP could be specifically released by phosphatidylinositol-specific phospholipase C (PIPLC), suggesting it is glycophosphatidylinositol-anchored to the plasma membrane. Therefore, a human growth hormone-PFAP fusion was constructed to locate the cleavage/attachment site. Immunofluorescent labeling and immunoblotting showed that Asn-477 is the cleavage/attachment site and the 25-residue peptide COOH-terminal to Asn-477 is removed during glycophosphatidylinositol anchoring. This research will hopefully pave the way to illustrate the role PFAP plays in calcium transportation related to pearl biomineralization. PMID:17624576

Xie, Li-Ping; Wu, Yuan-Tai; Dai, Yi-Ping; Li, Qing; Zhang, Rong-Qing

2007-01-01

189

Induction of rat alkaline phosphatase isozymes bearing a glycan-phosphatidylinositol anchor modified by in vivo treatment with a benzimidazole derivative linked to ethylbenzene.  

Science.gov (United States)

Serum alkaline phosphatase (ALP) is detected in soluble-form as a result of translocation from the membrane site by cleavage at the glycosyl-phosphatidylinositol moiety (GPI anchor). It is known that membrane-bound ALP (mALP) can be detected in serum in certain pathological and physiological conditions, and that it can be solubilized in vitro to soluble-ALP (sALP) by phosphatidylinositol-specific phospholipase C (PIPLC), phospholipase D, bile salt, detergent, etc. We observed a marked increase in ALP activity in the serum of rats given a benzimidazole derivative by gavage, and detected it as slow-migrating ALPs (SM-ALPs), which were mALP-like but resistant to PIPLC and n-butanol treatment on disc PAGE. On the other hand, ficin treatment made SM-ALPs shift to the sALP position. The molecular size of the SM-ALPs was smaller than that of sALP on sodium dodecyl sulphide-polyacrylamide slab-gel electrophoresis (SDS-PAGE), and immunoreactivity revealed the intestinal type. SM-ALPs were also detected in the duodenum and jejunum. The main sugar chain structure of SM-ALPs was the biantennary complex-type, which was coincided with intestinal sALP sugar chain. These results suggest that intestinal ALPs induced by the benzimidazole derivative were modified in their C-terminus or GPI anchor region and modification of this region may also participate in translocation into the bloodstream. PMID:11079373

Harada, T; Koyama, I; Sato, K; Komoda, T

2000-10-01

190

Potentiating role of IGFBP-2 on IGF-II-stimulated alkaline phosphatase activity in differentiating osteoblasts.  

Science.gov (United States)

The insulin-like growth factor (IGF) system plays an important role in the autocrine and paracrine regulation of bone formation and remodeling. The aim of this study was to evaluate the role of the autocrine IGF system during osteogenic differentiation in rat tibial osteoblasts (ROB) in culture. In this in vitro model, the stages of osteogenesis studied were S1, corresponding to the onset of alkaline phosphatase (AP) expression (days 0-3); S2, coincident with the peak of AP expression in differentiation culture conditions (days 4-6), and S3, corresponding to the onset of mineral deposition in the extracellular matrix (days 7-9). The results showed that conditioned medium of ROB contains greater amounts of IGF-II than IGF-I at all differentiation stages. Both peptides showed the highest concentrations on day 3 of differentiation (end of S1). All IGF-binding proteins (IGFBPs), except IGFBP-1 and -6, were detected, and IGFBP-2 was the most abundant IGFBP present in the conditioned media, and its degradation increased from S1 to S3. By semiquantitative RT-PCR, IGF-I and IGF-II were highly expressed on days 3 and 6, whereas IGFBP-2 was constantly expressed. We focused our study on the role of IGF-II and IGFBP-2 on the synthesis of AP, an early marker of osteoblast maturation. The results showed that a significant increase in AP expression was induced by IGF-II added to the differentiating osteoblasts continuously or in S1 but not in S2 or S3. IGFBP-2 was able to potentiate endogenous and exogenous IGF-II-dependent stimulation of AP activity, and its proteolytic degradation in late stages of osteogenesis (S2 and S3) was highly correlated with the increase of active matrix metalloproteinase-2 in the CM and with the decreased efficacy of IGF-II action. These data suggest that IGFBP-2, at nearly equimolar concentration with IGF-II, plays a potentiating role in IGF-II action on ROB differentiation in vitro. PMID:14665441

Palermo, Claudia; Manduca, Paola; Gazzerro, Elisabetta; Foppiani, Luca; Segat, Daniela; Barreca, Antonina

2004-04-01

191

Combined influence of temperature and metal ions on the level of activity of alkaline phosphatase of intestinal mucosa of Acipenseridae  

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Full Text Available The combined influence of divalent metal ions (Mn, Fe, Co, Ni, Cu and Zn and temperature on the level of alkaline phosphatase activity of the mucous membrane of the Acipenseridae is shown. The dependence of the response of the enzyme to the action of metal ions according to their position in the periodic table of chemical elements is presented. This dependence is kept during the change of incubation temperature as well, but only at low temperatures the activating effect of metals is maximum in the early period at high temperatures the inhibitory effects of metals is maximum at the end of the period.

Bednyakov Dmitriy Andreevich

2012-11-01

192

Pst I restriction fragment length polymorphism of the human placental alkaline phosphatase gene in normal placentae and tumors.  

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The structure of the human placental alkaline phosphatase gene from normal term placentae was studied by restriction enzyme digestion and Southern blot analysis using a cDNA probe to the gene for the placental enzyme. The DNA digests fall into three distinct patterns based on the presence and intensity of an extra 1.1-kilobase Pst I band. The extra 1.1-kilobase band is present in 9 of 27 placenta samples, and in 1 of these samples the extra band is present at double intensity. No polymorphism...

Tsavaler, L.; Penhallow, R. C.; Kam, W.; Sussman, H. H.

1987-01-01

193

Changes in Biliary (High-Molecular-Mass) and Liver Isoforms of Alkaline Phosphatase After Baboon-to-Human Liver Transplantation  

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We report a case of hyperphosphatasemia in a 35-year-old patient with hepatitis B who underwent an orthotopic xenogeneic liver transplant. Marked increases in total alkaline phosphatase (ALP; EC 3.1.3.1) activity began 5 days posttransplantation (six times human normal) and increased to ?17 times normal at day 11. Increased ALP persisted for >40 days and steadily increased to 75 times normal in the patient's last 30 days. Gel electrophoresis detected both liver (LALP) and biliary (high-mole...

Mercer, Donald; Tang, Mary; Marino, Ignazio R.; Demetris, Anthony; Fung, John; Starzl, Thomas; Warty, Vijay

1994-01-01

194

Use of an alkaline phosphatase-labeled synthetic oligonucleotide probe for detection of Campylobacter jejuni and Campylobacter coli.  

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A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to detect either 5 ng of C. jejuni DNA or 10(5) CFU of bacteria. The SNAP could also detect DNA extracted from 10(5) CFU in mock-infected stool samples. The SNAP detected C. jejuni and C. coli but showed no reactivity with C. laridis, C. fetus subsp. fetus, C. fetus subsp. venerealis, C. fennelliae,...

1990-01-01

195

The effect of rectal examination on serum acid phosphatase levels in benign and malignant prostatic disese  

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Serum acid phosphatase was measured in patients with enlarged benign and malignant prostate before and after rectal examination. Amongst the patients with benign glands, rectal examination did not produce any significant false elevation of the enzyme. Rectal examination, however, caused a rise in the enzyme level in a few untreated cancer patients and in cancer patients who had become refractory to hormonal therapy. This rise would help rather than mislead in the diagnosis of malignant prosta...

1982-01-01

196

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

Energy Technology Data Exchange (ETDEWEB)

We describe radioimmunoassay for human prostatic acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) 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 ..mu..g/liter and for 12 patients with advanced prostatic carcinoma between 100 and 500 ..mu..g/liter. The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group.

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

1978-11-01

197

Stability of lactate dehydrogenase, aspartate aminotransferase, alkaline phosphatase and tartrate resistant acid phosphatase in human saliva and gingival crevicular fluid in the presence of protease inhibitor  

Directory of Open Access Journals (Sweden)

Full Text Available The stability of aspartate aminotransferase (AST, lactate dehydrogenase (LDH, tartrate resistant acid phosphatase (TRAP and alkaline phosphatase (ALP activities from saliva and gingival crevicular fluid (GCF with and without the addition of protease inhibitor (PI at room temperature (RT; 25°C, 4°C and -20°C were investigated. AST, LDH, TRAP and ALP activities in saliva and GCF (n=9 with and without the addition of PI were assayed at 0 (control, 12, 24, 48, 72 h, one and two weeks. A paired t-test showed there were a significant differences (p<0.05 between LDH and TRAP activities in saliva, in the presence and without PI at all temperatures. ALP activity exhibited a significant difference in activity (p<0.05 in the presence and without PI at RT while no significant differences were observed at 4ºC and -20ºC. A significant difference (p<0.05 was observed in AST, LDH and TRAP activities (GCF at RT and 4ºC in the presence and without PI. We conclude that PI is essential for maintaining stable enzyme activities in saliva and GCF.

Kasim Nurfathiha Abu

2013-01-01

198

Factors Affecting Alkaline Phosphatase Activity of the Marine Cyanobacterium Lyngbya majuscula  

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Some environmental factors affecting phosphatase activity of the cyanobacterium Lyngbya majuscula found on the coral reefs in the Red Sea were investigated. Phosphatase activity of L. majuscula was restricted only to phosphomonoesterase (PMEase) without any detectable level of phosphodiesterase (PDEase) during all experiments. The maximum enzyme activity was obtained at 35°C (69.1 pNP ?mol mg-1 dry wt. h-1) and at pH 10 (89.1 mol pNP mg...

Al-shehri, Abdulrahman M.

2006-01-01

199

Maltol complexes of vanadium (IV) and (V) regulate in vitro alkaline phosphatase activity and osteoblast-like cell growth  

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Vanadium compounds have been found to possess insulin- and growth factor-mimetic effects. In consequence, these derivatives are potentially useful as effective oral therapeutic agents in diabetic patients. However, their use has been limited by various toxic side-effects and by the low solubility of different derivatives. Recently, vanadium complexes with maltol, a sugar used as a common food additive, have been synthesised and investigated in animals, showing possible insulin-mimetic effects with low toxic side-effects. In the present study we have investigated the effect of bis(maltolato)oxovanadium (IV) (BMOV) and bis(maltolato)dioxovanadium (V) (BMV) on bone cells in culture as well as their direct effect on alkaline phosphatase in vitro. A comparison was also made with the action of vanadate and vanadyl cation. Vanadium compounds regulated cell proliferation in a biphasic manner with similar potencies. Osteoblast differentiation, assessed by alkaline phosphatase activity, was found to be dose-dependent, with the inhibitory effect being stronger for vanadate and BMOV than for vanadyl and BMV. All vanadium compounds directly inhibited bovine intestinal ALP with a similar potency. Thus, maltol vanadium derivatives behave in a similar way to vanadate and vanadyl in osteoblast-like UMR 106 cells in culture. (orig.)

Barrio, D.A.; Braziunas, M.D. [Catedra de Bioquimica Patologica, Universidad Nacional de la Plata (Argentina); Etcheverry, S.B. [Catedra de Bioquimica Patologica, Universidad Nacional de la Plata (Argentina)]|[CEQUINOR, Facultad de Ciencias Exactas, Universidad Nacional de la Plata (Argentina); Cortizo, A.M. [CEQUINOR, Facultad de Ciencias Exactas, Universidad Nacional de la Plata (Argentina)

1997-12-31

200

Whole blood staining in suspension for nonspecific esterase and alkaline phosphatase analyzed with a Technicon H-1.  

Science.gov (United States)

We adapted previously published methods for nonspecific esterase and alkaline phosphatase staining of white blood cells in suspension for use on a Technicon H-1 hematology analyzer. The objective was to develop a semiautomated method using whole blood that could be employed on a large scale for hematology laboratory applications, including toxicology studies, measurement of neutrophil left shift, and cytochemical classification of myeloid leukemias. The nonspecific esterase method uses the pararosaniline stain, generating the unstable substrate from two stable precursors. Whole blood is added to the substrate plus dye mix. Next, acid lysis and fixation steps destroy red cells and stabilize the monocyte staining. The alkaline phosphatase stain employs a stable naphthyl phosphate substrate and fast blue B coupling dye. The red cells are lysed with a pH 10.3 propanediol buffer, and the white blood cells are then stabilized with formalin fixation. For both methods the staining is performed off-line, and the sample is then diluted with propanediol to match the refractive index of the sheath on the H-1 analyzer, before aspiration into the direct cytometry port. A cytogram of scattered versus absorbed light is obtained. The number of cells staining and the intensity of the stain can be quantified from the cytogram. PMID:1693116

Ross, D W; Bishop, C; Henderson, A; Kaplow, L

1990-01-01

 
 
 
 
201

Bacillus cereus Phosphopentomutase Is an Alkaline Phosphatase Family Member That Exhibits an Altered Entry Point into the Catalytic Cycle  

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Bacterial phosphopentomutases (PPMs) are alkaline phosphatase superfamily members that interconvert {alpha}-D-ribose 5-phosphate (ribose 5-phosphate) and {alpha}-D-ribose 1-phosphate (ribose 1-phosphate). We investigated the reaction mechanism of Bacillus cereus PPM using a combination of structural and biochemical studies. Four high resolution crystal structures of B. cereus PPM revealed the active site architecture, identified binding sites for the substrate ribose 5-phosphate and the activator {alpha}-D-glucose 1,6-bisphosphate (glucose 1,6-bisphosphate), and demonstrated that glucose 1,6-bisphosphate increased phosphorylation of the active site residue Thr-85. The phosphorylation of Thr-85 was confirmed by Western and mass spectroscopic analyses. Biochemical assays identified Mn{sup 2+}-dependent enzyme turnover and demonstrated that glucose 1,6-bisphosphate treatment increases enzyme activity. These results suggest that protein phosphorylation activates the enzyme, which supports an intermolecular transferase mechanism. We confirmed intermolecular phosphoryl transfer using an isotope relay assay in which PPM reactions containing mixtures of ribose 5-[{sup 18}O{sub 3}]phosphate and [U-{sup 13}C{sub 5}]ribose 5-phosphate were analyzed by mass spectrometry. This intermolecular phosphoryl transfer is seemingly counter to what is anticipated from phosphomutases employing a general alkaline phosphatase reaction mechanism, which are reported to catalyze intramolecular phosphoryl transfer. However, the two mechanisms may be reconciled if substrate encounters the enzyme at a different point in the catalytic cycle.

Panosian, Timothy D.; Nannemann, David P.; Watkins, Guy R.; Phelan, Vanessa V.; McDonald, W. Hayes; Wadzinski, Brian E.; Bachmann, Brian O.; Iverson, Tina M. (Vanderbilt)

2011-09-15

202

Bacillus cereus phosphopentomutase is an alkaline phosphatase family member that exhibits an altered entry point into the catalytic cycle.  

Science.gov (United States)

Bacterial phosphopentomutases (PPMs) are alkaline phosphatase superfamily members that interconvert ?-D-ribose 5-phosphate (ribose 5-phosphate) and ?-D-ribose 1-phosphate (ribose 1-phosphate). We investigated the reaction mechanism of Bacillus cereus PPM using a combination of structural and biochemical studies. Four high resolution crystal structures of B. cereus PPM revealed the active site architecture, identified binding sites for the substrate ribose 5-phosphate and the activator ?-D-glucose 1,6-bisphosphate (glucose 1,6-bisphosphate), and demonstrated that glucose 1,6-bisphosphate increased phosphorylation of the active site residue Thr-85. The phosphorylation of Thr-85 was confirmed by Western and mass spectroscopic analyses. Biochemical assays identified Mn(2+)-dependent enzyme turnover and demonstrated that glucose 1,6-bisphosphate treatment increases enzyme activity. These results suggest that protein phosphorylation activates the enzyme, which supports an intermolecular transferase mechanism. We confirmed intermolecular phosphoryl transfer using an isotope relay assay in which PPM reactions containing mixtures of ribose 5-[(18)O(3)]phosphate and [U-(13)C(5)]ribose 5-phosphate were analyzed by mass spectrometry. This intermolecular phosphoryl transfer is seemingly counter to what is anticipated from phosphomutases employing a general alkaline phosphatase reaction mechanism, which are reported to catalyze intramolecular phosphoryl transfer. However, the two mechanisms may be reconciled if substrate encounters the enzyme at a different point in the catalytic cycle. PMID:21193409

Panosian, Timothy D; Nannemann, David P; Watkins, Guy R; Phelan, Vanessa V; McDonald, W Hayes; Wadzinski, Brian E; Bachmann, Brian O; Iverson, Tina M

2011-03-11

203

Variations of alkaline phosphatase activity and P fractions in sediments of a shallow Chinese eutrophic lake (Lake Taihu)  

International Nuclear Information System (INIS)

The distribution of alkaline phosphatase activity (APA) and P fractions in sediment cores and the relationship between them were studied in a shallow Chinese freshwater lake (Lake Taihu). Sediment cores were collected from four sites, characterized by different degrees of eutrophication in June 2004. Sediment P was fractionated into Fe/Al-P, Ca-P, organic P (OP), inorganic P (IP) and total P (TP). The former two species made the largest contribution to the sediment P pool. Results show that trophic status and hydrological conditions have great impact on the APA of the sediments. The order of the APA in sediments was conjectured to be: macrophyte dominated lake > transitional lake > algal dominated lake. APA profiles follow a similar downcore decreasing trend. There was a positive relationship between the APA and the TP, IP. The multiple linear regression equation of the APA and P fractions is: APA = -97 + 0.768TP - 0.985Fe/Al-P. - Characteristics of the alkaline phosphatase activity and P fractions in sediments of different trophic status lake were studied in Lake Taihu

2007-11-01

204

Effect of vanadate, a potent alkaline phosphatase inhibitor, on 45Ca and 32P/sub i/ uptake by matrix vesicle-enriched fractions from chicken epiphyseal cartilage  

International Nuclear Information System (INIS)

Vanadate was tested in a hydroxyapatite-seeded ion uptake system to determine possible direct effects on mineral formation. The effect of vanadate on vesicle mineral ion uptake was complex; low dosages of vanadate (2-20 ?M) were stimulatory to Ca2+ uptake, but were inhibitory to P/sub i/. Higher dosages (>67 ?M) were inhibitory to both ions. The effect of vanadate on ion uptake was strongly influenced by the stage of vesicle loading; major effects were seen during the lag and early uptake phases, and minimal effects were seen in the terminal stages. Concentrations of vanadate highly inhibitory to vesicle ion uptake had minimal effects on ion accretion by a hydroxyapatite-seeded system. Inhibition of alkaline phosphatase activity by vanadate broadly paralleled inhibition of P/sub i/ and Ca2+ uptake; however, at low vanadate concentrations, inhibition of P/sub i/ uptake closely paralleled that of alkaline phosphatase. The data indicate that vanadate binds with high affinity to P/sub i/-loading sites, blocking initial P/sub i/ uptake. The close parallelism between inhibition of early P/sub i/ uptake and of alkaline phosphatase activity supports the concept that alkaline phosphatase is involved in P/sub i/ transport during the early stages of matrix vesicle ion loading. However, the fact that only about half of the P/sub i/ uptake was affected by vanadate, despite the progressive inhibition of alkaline phosphatase activity, indicates that alkaline phosphatase is not solely responsible for P/sub i/ uptake by the matrix vesicle-enriched fraction

1984-03-25

205

[Biochemical and histochemical studies of the alkaline phosphatase and adenosine triphosphatase levels in the lymph nodes of experimental animals after noise exposure].  

Science.gov (United States)

The authors examined biochemically and histochemically the activity of alkaline phosphatase and adenosinetriphosphatase in lymph nodules of experimental animals, living under the conditions of continuous noise action (3 and 5 months) at a level of 95 decibels A for 3 hours daily in the morning. There were phase changes in the activity of the examined enzymes, which revealed considerable stability even after stopping the contact of the organism with noise factor. The manifested inhibition of the activity of alkaline phosphatase and partly of adenosinetriphosphatase suggested that the cells of lymph tissue revealed disturbances in the metabolic processes, which caused exhaustion of their protective function. PMID:6454569

Softova, E; Zlateva, M; Iankova, T

1981-01-01

206

Factors Affecting Alkaline Phosphatase Activity of the Marine Cyanobacterium Lyngbya majuscula  

Directory of Open Access Journals (Sweden)

Full Text Available Some environmental factors affecting phosphatase activity of the cyanobacterium Lyngbya majuscula found on the coral reefs in the Red Sea were investigated. Phosphatase activity of L. majuscula was restricted only to phosphomonoesterase (PMEase without any detectable level of phosphodiesterase (PDEase during all experiments. The maximum enzyme activity was obtained at 35°C (69.1 pNP ?mol mg-1 dry wt. h-1 and at pH 10 (89.1 mol pNP mg-1 dry wt. h-1. The activity was markedly inhibited by Fe, Zn, Na, K and P ions at high concentrations (1 and 10 mM, but lower concentrations (0.01 and 0.1 mM enhanced this activity. The highest concentration of Ca and Mg ions (10 mM inhibited the enzyme activity, while lower concentrations stimulated this activity. Salinity had a marked effect on the enzyme activity, with a maximum obtained at 5‰.

Abdulrahman M. Al-Shehri

2006-01-01

207

Enzymatic labeling of a single chain variable fragment of an antibody with alkaline phosphatase by microbial transglutaminase.  

Science.gov (United States)

Functional cross-linking of a single chain Fv fragment of anti-hen egg-white lysozyme antibody (scFv) and alkaline phosphatase (AP) was explored using microbial transglutaminase (MTG) from Streptomyces mobaraensis. A specific peptidyl linker for MTG was genetically fused to the N-terminus of each protein and the resultant proteins were obtained separately by bacterial expression. The recombinant peptide-tagged scFv and AP were site-specifically cross-linked by MTG through the extra peptidyl linkers in vitro, which mainly yielded the heterodimer (i.e., scFv-AP conjugate). The enzymatic cross-linking reaction had little influence on either the antigen-binding ability of the scFv moiety or the enzymatic activity of the AP moiety of the conjugate, allowing use within an enzyme-linked immunosorbent assay. The results obtained suggest that the enzymatic approach with MTG facilitates the posttranslational construction of functional fusion proteins. PMID:15112292

Takazawa, Takeshi; Kamiya, Noriho; Ueda, Hiroshi; Nagamune, Teruyuki

2004-05-20

208

Effect of Zn(II) and Mg(II) on phosphohydrolytic activity of rat matrix-induced alkaline phosphatase.  

Science.gov (United States)

Rat matrix-induced alkaline phosphatase is an enzyme which requires magnesium and zinc ions for its maximal activity. Two Zn(II) ions and one Mg(II) ion are bound to each subunit of native dimeric enzyme. The presence of magnesium ion (10-100 microM) or zinc ion (7-20 nM) alone is sufficient to stimulate apoenzyme activity. However maximal activity (264 U/mg) requires the presence of both ions. Binding of Zn(II) ions to the Mg(II) binding site causes a strong inhibition of the apoenzyme while the binding of Mg(II) on Zn(II) binding site is not sufficient to stimulate PNPPase activity of the apoenzyme. Binding of both ions to the enzyme molecule did not change the apparent dissociation constant for PNPP hydrolysis. PMID:2611837

Ciancaglini, P; Pizauro, J M; Grecchi, M J; Curti, C; Leone, F A

1989-01-01

209

Fluorescence detection of adenosine-5'-triphosphate and alkaline phosphatase based on the generation of CdS quantum dots.  

Science.gov (United States)

We have developed an analytical method to detect adenosine-5'-triphosphate (ATP) and alkaline phosphatase (ALP) based on the generation of CdS quantum dots (QDs). We demonstrated that Cd(2+) cation reacts with S(2-) anion to generate fluorescent CdS QDs in the presence of some certain amount of ATP. With increase in the ATP concentration, the fluorescence intensity of CdS QDs was also enhanced. ATP can be converted into adenosine by the dephosphorylation of ALP, so that the generation of CdS QDs would be inhibited in the presence of ALP. Therefore, this novel analysis system could be applied to assay ATP and ALP based on the growth of fluorescent CdS QDs. PMID:24833001

Liu, Siyu; Wang, Xinyan; Pang, Shu; Na, Weidan; Yan, Xu; Su, Xingguang

2014-05-27

210

Investigating the kinetics of paramagnetic-beads linked alkaline phosphatase enzyme through microchannel resistance measurement in dielectric microchip.  

Science.gov (United States)

Real time monitoring of electrolyte resistance changes during hydrolysis of 4-nitrophenylphosphate (pNPP) by alkaline phosphatase (ALP) bound on paramagnetic-beads was performed into a small dielectric channel. The reaction kinetic fit with a non-competitive substrate-inhibition equation. Michaelis-Menten apparent constant, KM(app), was determined as 0.33±0.06mM and the maximum apparent rate, Vmax(app) as 98±5pMs(-1). The detection limits were 15fM for ALP and 0.75mM for pNPP. This miniaturized device constitutes a powerful tool for analysis of interaction between ligands. PMID:24613971

Faure, Mathilde; Sotta, Bruno; Gamby, Jean

2014-08-15

211

Nicotine inhibits collagen synthesis and alkaline phosphatase activity, but stimulates DNA synthesis in osteoblast-like cells  

International Nuclear Information System (INIS)

Use of smokeless tobacco is associated with various oral lesions including periodontal damage and alveolar bone loss. This study was performed to test the effects of nicotine on bone-forming cells at concentrations that occur in the saliva of smokeless tobacco users. Confluent cultures of osteoblast-like cells isolated from chick embryo calvariae were incubated for 2 days with nicotine added to the culture medium (25-600 micrograms/ml). Nicotine inhibited alkaline phosphatase in the cell layer and released to the medium, whereas glycolysis (as indexed by lactate production) was unaffected or slightly elevated. The effects on medium and cell layer alkaline phosphatase were concentration dependent with maximal inhibition occurring at 600 micrograms nicotine/ml. Nicotine essentially did not affect the noncollagenous protein content of the cell layer, but did inhibit collagen synthesis (hydroxylation of [3H]proline and collagenase-digestible protein) at 100, 300, and 600 micrograms/ml. Release of [3H]hydroxyproline to the medium was also decreased in a dose-dependent manner, as was the collagenase-digestible protein for both the medium and cell layer. In contrast, DNA synthesis (incorporation of [3H]thymidine) was more than doubled by the alkaloid, whereas total DNA content was slightly inhibited at 600 micrograms/ml, suggesting stimulated cell turnover. Morphologic changes occurred in nicotine-treated cells including rounding up, detachment, and the occurrence of numerous large vacuoles. These results suggest that steps to reduce the salivary concentration of nicotine in smokeless tobacco users might diminish damaging effects of this product on alveolar bone

1991-01-01

212

Nicotine inhibits collagen synthesis and alkaline phosphatase activity, but stimulates DNA synthesis in osteoblast-like cells  

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Use of smokeless tobacco is associated with various oral lesions including periodontal damage and alveolar bone loss. This study was performed to test the effects of nicotine on bone-forming cells at concentrations that occur in the saliva of smokeless tobacco users. Confluent cultures of osteoblast-like cells isolated from chick embryo calvariae were incubated for 2 days with nicotine added to the culture medium (25-600 micrograms/ml). Nicotine inhibited alkaline phosphatase in the cell layer and released to the medium, whereas glycolysis (as indexed by lactate production) was unaffected or slightly elevated. The effects on medium and cell layer alkaline phosphatase were concentration dependent with maximal inhibition occurring at 600 micrograms nicotine/ml. Nicotine essentially did not affect the noncollagenous protein content of the cell layer, but did inhibit collagen synthesis (hydroxylation of ({sup 3}H)proline and collagenase-digestible protein) at 100, 300, and 600 micrograms/ml. Release of ({sup 3}H)hydroxyproline to the medium was also decreased in a dose-dependent manner, as was the collagenase-digestible protein for both the medium and cell layer. In contrast, DNA synthesis (incorporation of ({sup 3}H)thymidine) was more than doubled by the alkaloid, whereas total DNA content was slightly inhibited at 600 micrograms/ml, suggesting stimulated cell turnover. Morphologic changes occurred in nicotine-treated cells including rounding up, detachment, and the occurrence of numerous large vacuoles. These results suggest that steps to reduce the salivary concentration of nicotine in smokeless tobacco users might diminish damaging effects of this product on alveolar bone.

Ramp, W.K.; Lenz, L.G.; Galvin, R.J. (Univ. of Louisville, KY (USA))

1991-05-01

213

Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots  

Scientific Electronic Library Online (English)

Full Text Available SciELO Mexico | Language: English Abstract in spanish Dos isoformas de fosfatasas obtenidas de raíz de frijol (Phaseolus vulgaris L.) mostraron un incremento en la actividad en respuesta a la deficiencia de fosfato. Una de ellas (APIII) se purificó a través de una cromatografía de intercambio iónico y una electroforesis preparativa. La masa molecular e [...] stimada para APIII fue de 35 kDa tanto por SDS-PAGE como por filtración molecular, sugiriendo que la enzima activa es monomérica. APIII se clasificó como una fosfatasa alcalina basada en sus requerimientos de pH 8 para catálisis. Esta enzima es activa sobre un amplio espectro de sustratos como polifosfato, glucose 1-fosfato y fosfoenolpiruvato, aunque muestra preferencia por pirofosfato. Su actividad se inhibe completamente por molibdato, vanadato y fosfato, aunque es inhibida parcialmente por fluoruro. Aún cuando los cationes divalentes no fueron escenciales para su actividad, la hidrólisis de pirofosfato se incrementó notablemente en presencia de Mg2+. Abstract in english 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 ma [...] ss 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 phosphoenol-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 pyrophosphatase activity of this enzyme, the hydrolysis of pyrophosphate increased substantially in the presence of Mg2+.

Lorena, Morales; Natalia, Gutiérrez; Vanessa, Maya; Carmen, Parra; Eleazar, Martínez-Barajas; Patricia, Coello.

214

Assay Format as a Critical Success Factor for Identification of Novel Inhibitor Chemotypes of Tissue-Nonspecific Alkaline Phosphatase from High-Throughput Screening  

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The tissue-nonspecific alkaline phosphatase (TNAP) isozyme is centrally involved in the control of normal skeletal mineralization and pathophysiological abnormalities that lead to disease states such as hypophosphatasia, osteoarthritis, ankylosis and vascular calcification. TNAP acts in concert with the nucleoside triphosphate pyrophosphohydrolase-1 (NPP1) and the Ankylosis protein to regulate the extracellular concentrations of inorganic pyrophosphate (PPi), a potent inhibitor of mineralizat...

Chung, Thomas D. Y.; Eduard Sergienko; José Luis Millán

2010-01-01

215

The effect of 1,25-dihydroxycholecalciferol on the multiple forms of alkaline phosphatase and the sialic acid incorporation into microsomes of chick duodenum  

International Nuclear Information System (INIS)

Polyacrylamide disc gel electrophoresis of n-butanol solubilized alkaline phosphatase from chick duodenum revealed that the change of alkaline phosphatase induced by 1,25-(OH)2D3 involved the transformation of desialoenzyme to sialoenzyme. The initial stimulation by 1,25-(OH)2D3 of the incorporation of sialic acid into duodenal microsomes corresponded with the initial increase in calcium absorption. After this initial stimulation, there was a rapid decline in sialic acid incorporation into microsomes decreasing below control levels at 24 hr. Calcium concentration in the microsomes followed a pattern similar to the incorporation of sialic acid into microsomes. The depressed sialic acid incorporation was reversed by the addition of calcium in vitro. These results suggest that the initial action of 1,25-(OH)2D3 is to change the membrane permeability to calcium and to change the subcellular distribution of calcium in the small intestine. The accumulated calcium in the microsomes then stimulates the sialic acid incorporation into desialoenzyme. This results in the changes of isozyme pattern of alkaline phosphatase, viz, the transformation of desialoenzyme to sialoenzyme. The transformed alkaline phosphatase might be one of the factors involved more directly in the regulation of calcium transport in intestine. (auth.)

1977-01-01

216

CONTROL OF ALKALINE PHOSPHATASE ACTIVITY IN C3H10T1/2 CELLS: ROLE OF RETINOIC ACID AND CELL DENSITY  

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The enzyme alkaline phosphatase (AP) has been shown to be lost or inappropriately expressed during carcinogenesis in some tissues. ecause retinoic acid (RA) appears to play a role in the normal regulation of the enzyme (RA up-regulates AP in a variety of cell types) we have sugge...

217

Kinetic comparison of tissue non-specific and placental human alkaline phosphatases expressed in baculovirus infected cells: application to screening for Down's syndrome  

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Full Text Available Abstract Background In humans, there are four alkaline phosphatases, and each form exibits a characteristic pattern of tissue distribution. The availability of an easy method to reveal their activity has resulted in large amount of data reporting correlations between variations in activity and illnesses. For example, alkaline phosphatase from neutrophils of mothers pregnent with a trisomy 21 fetus (Down's syndrome displays significant differences both in its biochemical and immunological properties, and in its affinity for some specific inhibitors. Results To analyse these differences, the biochemical characteristics of two isozymes (non specific and placental alkaline phosphatases were expressed in baculovirus infected cells. Comparative analysis of the two proteins allowed us to estimate the kinetic constants of denaturation and sensitivity to two inhibitors (L-p-bromotetramisole and thiophosphate, allowing better discrimination between the two enzymes. These parameters were then used to estimate the ratio of the two isoenzymes in neutrophils of pregnant mothers with or without a trisomy 21 fetus. It appeared that the placental isozyme represented 13% of the total activity of neutrophils of non pregnant women. This proportion did not significantly increase with normal pregnancy. By contrast, in pregnancies with trisomy 21 fetus, the proportion reached 60–80% of activity. Conclusion Over-expression of the placental isozyme compared with the tissue-nonspecific form in neutrophils of mother with a trisomy 21 fetus may explain why the characteristics of the alkaline phosphatase in these cells is different from normal. Application of this knowledge could improve the potential of using alkaline phosphatase measurements to screen for Down's syndrome.

Grozdea Jean J

2002-01-01

218

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

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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...

Vijayaprasad K; Ravichandra K; Vasa A.A.K; Suzan S

2010-01-01

219

Formation of a vitamin B-12-serum complex on heating at alkaline pH  

International Nuclear Information System (INIS)

The binding of vitamin B-12 to serum proteins during heating at alkaline pH was investigated by gel filtration of serum supplemented with cyano[57Co]-cobalamin. Heating for 5 min at 1000C destroyed most of the vitamin B-12 binding activity of serum but, with further heating, the vitamin B-12 became incorporated into a complex that did not correspond in molecular size to the original vitamin B-12 binding proteins. Radioassay of vitamin B-12 in heated serum showed correspondingly first an increase then a progressive decrease in the apparent vitamin B-12 level suggesting that, on heating, vitamin B-12 was initially released then subsequently complexed by the serum. The formation of complexed vitamin B-12 was abolished by the presence of the reducing agent dithiothreitol during the heating step. (Auth.)

1979-04-16

220

AVERRHOA CARAMBOLA (STAR FRUIT) INDUCES HEPATIC ALKALINE PHOSPHATASE ACTIVITY IN RATS  

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The objective of the study is to examine the in vivo effect of star fruit juice at different storage conditions on the activity of alkaline phosphates in female Sprague Dawley (SD) rats. A total of 20 healthy female rats weighing 180-200g were used for the experiment. All animals were divided into four groups with five animals per group (n=5). First control was served as control group. Second, third and forth groups were orally treated with a single dose daily of freshly ...

Chin J. H.; Teh C. C.; Khoo, Z. Y.; Shamala F.

2011-01-01

 
 
 
 
221

Identification of the genetic locus for the structural gene and a new regulatory gene for the synthesis of repressible alkaline phosphatase in Saccharomyces cerevisiae  

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Two lines of evidence showed that the PHO8 gene encodes the structure of repressible, nonspecific alkaline phosphatase in Saccharomyces cerevisiae: (I) the enzyme produced by a temperature-sensitive pho8 mutant at the permissive temperature (25/sup 0/C) was more thermolabile than that of the wild-type strain, and (II) the PHO8 gene showed a gene dosage effect on the enzyme activity. The pho8 locus has been mapped on chromosome IV, 8 centimorgans distal to rna3. A new mutant carrying the pho9 gene was isolated which lacks repressible alkaline phosphatase, but has the normal phenotype for the synthesis of repressible acid phosphatase. The pho9 gene segregated independently of all known pho-regulatory genes and did not show the gene dosage effect on repressible alkaline phosphatase activity. The pho9/pho9 diploid hardly sporulated and showed no commitment to intragenic recombination when it was inoculated on sporulation medium. Hence the pho9 mutant has a phenotype similar to the pep4 mutant, which was isolated as a pleiotropic mutant with reduced levels of proteinases A and B carboxypeptidase Y. An allelism test indicated that pho9 and pep4 are allelic.

Kaneko, Y.; Toh-e, A.; Oshima, Y.

1982-02-01

222

Stimulation by parathyroid hormone of 45Ca2+ uptake in osteoblast-like cells: Possible involvement of alkaline phosphatase  

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We have investigated the actions of human PTH [hPTH-(1-34)] on the association of 45Ca2+ with two human (SaOS-2 and MG-63) and two rat (ROS 17/2.8 and UMR-106) osteoblast-like cell types. In SaOS-2 cells, hPTH-(1-34) binds to specific membrane receptors to activate adenylate cyclase. Treatment of SaOS-2 cells with hPTH-(1-34) resulted in an increase in 45Ca2+ uptake, in a dose-dependent fashion, up to 2- to 4-fold above control values. The increase was first evident at 10 min and persisted for at least 30 min. Treatment with nimodipine, a calcium channel antagonist, was without effect on the stimulatory action of PTH. A similar enhancement of cell-associated 45Ca2+ was observed when the cells were incubated with vasoactive intestinal peptide, which acts via different receptors to activate adenylate cyclase in SaOS-2 cells. Treatment with (Bu)2cAMP also induced an increase in cell-associated 45Ca2+. Pretreatment of SaOS-2 cells with hPTH-(1-34) for 4 h, which induced homologous desensitization to a second challenge with the same peptide for stimulation of cAMP production, did not attenuate the further enhancement of cell-associated 45Ca2+ by a second treatment with hPTH-(1-34). We then examined a possible relationship between alkaline phosphatase (ALPase) and 45Ca2+ uptake. SaOS-2 cells contained high levels of alkaline phosphatase activity and continuously released the enzyme into the medium. Release was enhanced by treatment with hPTH-(1-34) for 10 min. Incubation of cells with levamisole (an inhibitor of the liver/bone/kidney type of ALPase) resulted in a rapid decrease in basal and PTH-stimulated 45Ca2+ uptake, while treatment with L-Phe-Gly-Gly was without effect. Treatment of the cells with ALPase (bovine kidney) enhanced 45Ca2+ uptake. In MG-63 cells, a stimulatory effect of hPTH-(1-34) on cell-associated 45Ca2+ was also observed; however, hPTH-(1-34) did not stimulate cAMP production in MG-63 cells

1990-01-01

223

Comparison of Salivary Ion Activity Product for Hydroxyapatite (IPHA, Alkaline Phosphatase and Buffering Capacity of Adults According to Age and Caries Severity  

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Full Text Available Statement of Problem: Tooth caries is influenced by different biochemical characteristics of saliva. As hydroxyapatite is the main component of enamel, salivary ion activity product for hydroxyapatite (IPHA as well as alkaline phosphatase may be attributed to dental caries. Purpose: The aim of the present study was to compare salivary buffering capacity, alkaline phosphatase and IPHA of adults according to the dental caries and age. Materials and Method: One hundred and twenty 19 to 44 years old male individuals were divided into four groups according to the dental caries rate and age: group 1: 19-35 years old low dental caries (DMFT <5; group 2: 19-35 years old high dental caries (DMFT 5<; group 3: 35-44 years old low dental caries (DMFT <11 and 35-44 years old high dental caries (DMFT 11<. Five millilitre of unstimulated saliva was collected, and then buffering capacity, the level of alkaline phosphatase activity and IPHA was determined for each sample. Data was analyzed by soft ware SPSS using two-way ANOVA, Friedman and Mann-Whitney tests.Results: Mean and standard deviation of buffering capacity of group 1 to 4 was 2.66±0.54, 2.64±0.56, 2.70±0.70 and 2.26±0.82, respectively. The difference was not significance (p= 0. 305. Mean and standard deviation of alkaline phosphatase activity of group 1 to 4 was 5.82±2.91, 5.30±1.52, 4.77±1.82 and 4.55±1.61, respectively. There was no significant difference (p= 0.692. Mean and standard deviation of IPHA of group 1 to 4 was 29.39±0.61, 29.51±0.76, 29.14±0.56 and 29.75±0.75, respectively. The difference was significant (p= 0.049.Conclusion: Based on the results of the present study, buffering capacity and the level of alkaline phosphatase couldn’t affect dental caries, independently. However, the higher value of IPHA may be attributed to the higher dental caries rate. Ageing decreases alkaline phosphatase activity.

Vahedi M.

2012-12-01

224

Alkaline Phosphatase Activity : an overlooked player on the phosphate behavior in macrotidal estuaries  

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The non-conservative behavior of phosphate within the estuarine salinity gradient is essentially assigned to physico-chemical processes, such as desorption at low salinity and to benthic exchanges. Microbial phosphatase activity (APA), generally related to phosphate deficiency, is seldom studied in phosphate rich estuarine waters. In order to address the impact of microbial activity (bacterial abundance, production BSP, APA) on phosphate behavior, we studied these activities on a seasonal basis within the salinity gradient of two macrotidal estuaries presenting different levels of suspended solids. Whatever the season the Charente estuary is characterized by high levels of Suspended Particulate Matter (SPM > 1g.L-1), particularly in the Maximum Turbidity Zone (MTZ) located at the 5-10 psu. In this area characterized by high BSP and APA there is a significant increase of PO4 levels especially during summer. In the Aulne estuary the particle load is significantly lower (1/10) but high BSP and APA are equally recorded. In the highly turbid waters of the Charente estuary, active phytoplankton is virtually absent as pheopigments constitute up to 80% of the total pigments, particularly in the MTZ, therefore APA may essentially have a bacterial origin. In the Aulne estuary attached bacteria are dominant, both in numbers and production, and their distribution along the haline gradient perfectly follows those of APA and phosphate levels. These observations, associated with the very close relationships observed between APA, SPM and BSP, suggest that APA derive mainly from bacterial (attached) origin and operate at the expense of particulate phosphorus and hence contribute to PO4 regeneration, especially in spring and summer. Finally, as APA increased as PO4, whereas the reverse is observed in both fresh and marine waters, an original scheme for APA regulation, related to the large dominance of attached bacteria can be described for the estuarine waters.

Delmas, Daniel; Labry, Claire; Youenou, Agnes; Quere, Julien; Auguet, Jean Christophe; Montanie, Helene

2014-05-01

225

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

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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.

McKee, Marc D.

2008-09-01

226

Alkaline phosphatase as a reporter of sigma(S) levels and rpoS polymorphisms in different E. coli strains.  

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sigma(S) is responsible for the transcriptional regulation of genes related to protection against stresses and bacterial survival and it accumulates in the cell under conditions of stress, such as nutrient limitation. An increase in the levels of sigma(S) causes a reduction in the expression of genes that are transcribed by RNA polymerase associated with the principal sigma factor, sigma(70). phoA, that encodes alkaline phosphatase (AP) is expressed under phosphate shortage conditions, and is also repressed by sigma(S). Here we show that in a Pi-limited chemostat, accumulation of rpoS mutations is proportional to the intrinsic level of sigma(S) in the cells. Acquisition of mutations in rpoS relieves repression of the PHO genes. We also devised a non-destructive method based on the rpoS effect on AP that differentiates between rpoS (+) and rpoS mutants, as well as between high and low-sigma(S) producers. Using this method, we provide evidence that sigma(S) contributes to the repression of AP under conditions of Pi excess and that AP variation among different strains is at least partly due to intrinsic variation in sigma(S) levels. Consequently, a simple and non-destructive AP assay can be employed to differentiate between strains expressing different levels of sigma(S) on agar plates. PMID:17665171

Spira, Beny; Ferenci, Thomas

2008-01-01

227

Arginine Coordination in Enzymatic Phosphoryl Transfer: Evaluation of the Effect of Arg166 Mutations in Escherichia Coli Alkaline Phosphatase  

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Arginine residues are commonly found in the active sites of enzymes catalyzing phosphoryl transfer reactions. Numerous site-directed mutagenesis experiments establish the importance of these residues for efficient catalysis, but their role in catalysis is not clear. To examine the role of arginine residues in the phosphoryl transfer reaction, we have measured the consequences of mutations to arginine 166 in Escherichia coli alkaline phosphatase on hydrolysis of ethyl phosphate, on individual reaction steps in the hydrolysis of the covalent enzyme-phosphoryl intermediate, and on thio substitution effects. The results show that the role of the arginine side chain extends beyond its positive charge, as the Arg166Lys mutant is as compromised in activity as Arg166Ser. Through measurement of individual reaction steps, we construct a free energy profile for the hydrolysis of the enzyme-phosphate intermediate. This analysis indicates that the arginine side chain strengthens binding by ?3 kcal/mol and provides an additional 1-2 kcal/mol stabilization of the chemical transition state. A 2.1 (angstrom) X-ray diffraction structure of Arg166Ser AP is presented, which shows little difference in enzyme structure compared to the wild-type enzyme but shows a significant reorientation of the bound phosphate. Altogether, these results support a model in which the arginine contributes to catalysis through binding interactions and through additional transition state stabilization that may arise from complementarity of the guanidinum group to the geometry of the trigonal bipyramidal transition state

2008-07-22

228

Relation of fatty acid composition in lead-exposed mallards to fat mobilization, lipid peroxidation and alkaline phosphatase activity  

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The increase of n-6 polyunsaturated fatty acids (PUFA) in animal tissues has been proposed as a mechanism of Pb poisoning through lipid peroxidation or altered eicosanoids metabolism. We have studied fatty acid (FA) composition in liver and brain of mallards (Anas platyrhynchos) feeding for three weeks on diets containing combinations of low or high levels of vitamin E (20 or 200 UI/kg) and Pb (0 or 2 g/kg). Saturated FA, n-6 PUFA and total concentrations of FA were higher in livers of Pb-exposed mallards, but not in their brains. The percentage of n-6 PUFA in liver and brain was slightly higher in Pb-exposed mallards. The increase of n-6 PUFA in liver was associated with increased triglycerides and cholesterol in plasma, thus could be in part attributed to feed refusal and fat mobilization. The hepatic ratios between adrenic acid (22:4 n-6) and arachidonic acid (20:4 n-6) or between adrenic acid and linoleic acid (18:2 n-6) were higher in Pb exposed birds, supporting the existing hypothesis of increased fatty acid elongation by Pb. Among the possible consequences of increased n-6 PUFA concentration in tissues, we found increased lipid peroxidation in liver without important histopathological changes, and decreased plasma alkaline phosphatase activity that may reflect altered bone metabolism in birds.

Mateo, R.; Beyer, W.N.; Spann, J.W.; Hoffman, D.J.

2003-01-01

229

Characterization of the tryptophan residues of Escherechia coli alkaline phosphatase by phosphorescence and optically detected magnetic resonance spectroscopy.  

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The phosphorescence and zero field optically detected magnetic resonance (ODMR) of the tryptophan (Trp) residues of alkaline phosphatase from Escherechia coli are examined. Each Trp is resolved optically and identified with the aid of the W220Y mutant and the terbium complex of the apoenzyme. Trp(109), known from earlier work to be the source of room-temperature phosphorescence (RTP), emits a highly resolved low-temperature phosphorescence (LTP) spectrum and has the narrowest ODMR bands observed thus far from any protein site, revealing a uniquely homogeneous local environment. The decay kinetics of Trp(109) at 1.2 K reveals that the major triplet population (70%) undergoes inefficient crystallike spin-lattice relaxation by direct interaction with lattice phonons, the remainder being relaxed efficiently by local disorder modes. The latter population is smaller than is typical for protein sites, suggesting an unusual degree of local rigidity and order consistent with the long-lived RTP. Trp(220) emits a broader LTP spectrum originating to the blue of Trp(109). It has typically broad ODMR bands consistent with local heterogeneity. The LTP of Trp(268) has an ill-defined origin blue shifted relative to Trp(220) and ODMR frequencies consistent with a greater degree of solvent exposure. Trp(268) has noticeable dispersion of its decay kinetics, consistent with quenching at the triplet level by a nearby disulfide residue. PMID:11732924

Ghosh, S; Misra, A; Ozarowski, A; Stuart, C; Maki, A H

2001-12-11

230

Sub-cellular localisation of alkaline phosphatase activity in the cytoplasm of tammar wallaby (Macropus eugenii) neutrophils and eosinophils.  

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Alkaline phosphatase (ALP) has been used in studies of neutrophil morphology and function as a marker for identifying different granule populations. In human neutrophils, ALP is found within secretory vesicles, a rapidly mobilisable vesicle population important for upregulating membrane receptors during early activation. Intra-cellular ALP activity in the heterophils of rabbits and guinea pigs, in contrast, is found only in secondary granules. The neutrophils and eosinophils of tammar wallabies (Macropus eugenii) have previously been reported to contain large amounts of ALP activity when stained using routine cytochemical techniques. To define the subcellular location of ALP in this species, cell suspensions were examined using cerium chloride cytochemistry and transmission electron microscopy (TEM). ALP was found in 2 distinct cytoplasmic compartments. One compartment displayed morphology consistent with a subpopulation of secondary granules while a second tubulo-vesicular population appeared similar to the secretory vesicles of human neutrophils. Thin tubular vesicles containing ALP were also identified within the cytoplasm of tammar wallaby eosinophils. Large numbers of ALP-containing vesicles have not been recognised previously in eosinophils and this may represent a novel cytoplasmic compartment. In both cell types, ALP-containing structures showed alteration in morphology following stimulation with N-formyl-Met-Leu-Phe (fMLP) and PMA. PMID:21596444

Hulme-Moir, K Lisa; Clark, Phillip

2011-07-15

231

Influence of dissolved humic materials on carbon assimilation and alkaline phosphatase activity in natural algal-bacterial assemblages  

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Mixed natural assemblages of algae and bacteria exhibited lower rates of /sup 14/C assimilation and high rates of dissimilation of recent photosynthate when amended with low concentrations of unfractioned dissolved humic material (DHM). The extent of the inhibition or stimulation was greatest in the smaller (1-5 ..mu..m) assemblage particles. In different algal-bacterial assemblages, additions of DHM markedly enhanced community alkaline phosphatase activity (APA), particularly under low light regimes. DHM of low apparent molecular weight was much more stimulatory to both /sup 14/C assimilation and APA than DHM of high apparent molecular weight, supporting the belief that DHM molecular weight is an important determinant of DHM interactive capacity. Addition of phosphate enhanced the disparity in rates of /sup 14/C assimilation of samples incubated under low and high light regimes, increased the rates of /sup 14/C assimilation, and depressed APA. There were indications of interactions between DHM and phosphorus in several experiments. Two hypotheses were invoked to explain increases in APA in response to DHM.

Stewart, A.J.; Wetzel, R.G.

1982-01-01

232

Expression of SEAP (secreted alkaline phosphatase) by baculovirus mediated transduction of HEK 293 cells in a hollow fiber bioreactor system.  

Science.gov (United States)

A BacMam baculovirus was designed in our laboratory to express the reporter protein secreted alkaline phosphatase (SEAP) driven by the immediate early promoter of human cytomegalovirus promoter (CMV). In vitro tests have been carried out using this recombinant baculovirus to study the secreted protein in two cell lines and under various culture conditions. The transductions were carried out on two commonly used mammalian cell lines namely the human embryonic kidney (HEK 293A) and Chinese hamster ovary (CHO-K1). Initial studies clearly demonstrated that the transient expression of SEAP was at least 10-fold higher in the HEK 293 cells than the CHO cells under equivalent experimental conditions. Factorial design experiments were done to study the effect of different parameters such as cell density, MOI, and the histone deacetylase inhibitor, trichostatin A concentration. The multiplicity of infection (MOI) and the cell density were found to have the most impact on the process. The enhancer trichostatin A also showed some positive effect. The production of secreted protein in a batch reactor was studied using the Wave disposable bioreactor system. A semi-continuous perfusion process was developed to extend the period of gene expression in mammalian cells using a hollow fiber bioreactor system (HFBR). The growth of cells and viability in both systems was monitored by offline analyses of metabolites. The expression of recombinant protein could be maintained over an extended period of time up to 30 days in the HFBR. PMID:18499293

Jardin, B A; Zhao, Y; Selvaraj, M; Montes, J; Tran, R; Prakash, S; Elias, C B

2008-06-30

233

Arginine Coordination in Enzymatic Phosphoryl Transfer: Evaluation of the Effect of Arg166 Mutations in Escherichia Coli Alkaline Phosphatase  

Energy Technology Data Exchange (ETDEWEB)

Arginine residues are commonly found in the active sites of enzymes catalyzing phosphoryl transfer reactions. Numerous site-directed mutagenesis experiments establish the importance of these residues for efficient catalysis, but their role in catalysis is not clear. To examine the role of arginine residues in the phosphoryl transfer reaction, we have measured the consequences of mutations to arginine 166 in Escherichia coli alkaline phosphatase on hydrolysis of ethyl phosphate, on individual reaction steps in the hydrolysis of the covalent enzyme-phosphoryl intermediate, and on thio substitution effects. The results show that the role of the arginine side chain extends beyond its positive charge, as the Arg166Lys mutant is as compromised in activity as Arg166Ser. Through measurement of individual reaction steps, we construct a free energy profile for the hydrolysis of the enzyme-phosphate intermediate. This analysis indicates that the arginine side chain strengthens binding by {approx}3 kcal/mol and provides an additional 1-2 kcal/mol stabilization of the chemical transition state. A 2.1 {angstrom} X-ray diffraction structure of Arg166Ser AP is presented, which shows little difference in enzyme structure compared to the wild-type enzyme but shows a significant reorientation of the bound phosphate. Altogether, these results support a model in which the arginine contributes to catalysis through binding interactions and through additional transition state stabilization that may arise from complementarity of the guanidinum group to the geometry of the trigonal bipyramidal transition state.

O' Brien, P.J.; Lassila, J.K.; Fenn, T.D.; Zalatan, J.G.; Herschlag, D.

2009-05-22

234

CONSUMPTION OF PHOSPHORUS AND PRODUCTION OF ALKALINE PHOSPHATASE DUR-ING GROWTH OF Streptomyces coelicolor IN A RICH MEDIUM  

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Full Text Available The growth of Strteptomyces coelicolor in a complex medium containing yeast extract, malt extract and glucose exhibited a biphasic mode of biomass accumulation. The first phase was associated with rapid biomass formation, phosphorus utilisation and lack of pigment production. The second stage was marked by slower growth and pigment formation. The demarcation between these phases appeared to result from the depletion of phosphorus in the media, which in turn allowed the red prodigiosin-like pigment to be expressed. Biomass formation appeared to involve the accumulation of phosphorus in the cells reaching a maximum level of 3.7 % of the dry weight. The depletion of phosphorus provoked the increase in alkaline phosphatase activity, which in turn produced a transient release of phosphorus into the medium, presumably from stored cellular phosphorus. Although glucose was consumed during growth, it did not constitute the only carbon source as the appearance of significant amounts of ammonium ions in the broth indicated deamination of amino compounds.

Ismini Nakouti and Glyn Hobbs

2012-02-01

235

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

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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 ?(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. PMID:23174485

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

2013-04-01

236

Prognostic significance of acid and alkaline phosphatases in head and neck cancer during radiotherapy  

International Nuclear Information System (INIS)

The present study is an attempt to observe the prognostic value of ACP and ALP in head and neck cancer patients. This study has been carried out on 30 patients with proven malignancies of head and neck cancer stage II, III and IV, at Shree Bhagwan Mahaveer Cancer Hospital and Research Center, Jaipur. Patients of head and neck cancers were classified according to UICC classification. The blood samples were collected and serum ACP and ALP levels were estimated at three intervals during treatment i.e. before therapy, mid therapy (25-30 Gt exposure) and after completion (50-70 Gt exposure) of radiotherapy. The levels of levels of ACP and ALP before therapy in head and neck were higher which later declined during treatment and continued decreasing till the completion of the therapy

2004-12-01

237

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

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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 ...

Benjamin Nnamdi Okolonkwo; Edna Ogechi Nwachuku

2013-01-01

238

Comparación del ultramicrométodo inmunocitoquímico (UMICIQ con el de la fosfatasa alcalina-anti fosfatasa alcalina (APAAP para la cuantificación de subpoblaciones linfocitarias T Comparison of the immunocytochemical ultramicromethod and the alkaline phosphatase - anti-alkaline phosphatase method for the quantification of T lymphocyte subsets  

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Full Text Available Se realizó un estudio comparativo entre el método inmunoenzimático que emplea el complejo fofatasa alcalina-anti fofatasa alcalina (APAAP y el ultramicrométodo inmunocitoquímico (UMICIQ utilizado para la detección de marcadores antigénicos y cuantificación de subpoblaciones de linfocitos T. Se estudiaron los antígenos celulares CD3, CD4 y CD8 en 30 individuos adultos supuestamente sanos. Al compararse los resultados por ambas técnicas, se encontraron diferencias estadísticamente significativas para una p A comparative study was conducted on the immunoenzymatic method based on the alkaline phosphatase - anti-alkaline phosphatase complex and the immunocytochemical ultramicromethod used for detecting antigen markers and for the quantification of T-lymphocyte subsets Cell antigens CD3, CD4 and CD8 from 30 apparently healthy adults were studied. When comparing the outcome of both techniques, statistically significant differences were found, (p< 0,05. It was concluded that although the diagnostic efficiency of both methods are similar, the alkaline phosphatase - anti-alkaline phosphatase method is quicker, more economical and less laborious than the immunocytochemical ultramicromethod, so it is recommended as a procedure of choice for these cell studies

Beatriz Socarrás Ferrer

2002-04-01

239

The effect of irradiation on the mRNA expression of type I collagen and alkaline phosphatase in the MC3T3-E1 osteoblastic cell line  

International Nuclear Information System (INIS)

To investigate the effects of irradiation on the phenotypic expression of the MC3T3-E1 osteoblastic cell line, particularly an the expression of type I collagen and alkaline phosphatase mRNA. Cells were irradiated with a single dose of 0.5, 1, 2, 4, and 8 Gy at a dose rate of 5.38 Gy/min using a cesium 137 irradiator. The specimens were then harvested and RNA extraction was carried out at 1 and 3 days after irradiation. The extracted RNA strands were reverse-transcribed and the resulting cDNA fragments were amplified by PCR. The irradiated cells demonstrated a dose-dependent increase in type I collagen mRNA expression relative to the control group, with a maximum level of type I collagen mRNA expression occurring at 8 Gy. The degree of type I collagen mRNA expression increased significantly at 1 day after irradiation, but little differences were found between the control group and at the 3rd day. The amount of alkaline phosphatase mRNA expression increased significantly at 1 and 3 days after irradiation in the 1 Gy exposed group compared with the control group. The amount of type I collagen and alkaline phosphatase mRNA expression increased significantly 1 day after irradiation when compared with the control group.

2003-03-01

240

Detection of prostatic cancer by solid-phase radioimmunoassay of serum prostatic acid phosphatase  

International Nuclear Information System (INIS)

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

1977-12-22

 
 
 
 
241

Ovarian cancer stem-like cells show induced translineage-differentiation capacity and are suppressed by alkaline phosphatase inhibitor.  

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Spheroid formation is one property of stem cells-such as embryo-derived or neural stem cells-that has been used for the enrichment of cancer stem-like cells (CSLCs). However, it is unclear whether CSLC-derived spheroids are heterogeneous or whether they share common embryonic stemness properties. Understanding these features might lead to novel therapeutic approaches. Ovarian carcinoma is a deadly disease of women. We identified two types of spheroids (SR1 and SR2) from ovarian cancer cell lines and patients' specimens according to their morphology. Both types expressed stemness markers and could self-renew and initiate tumors when a low number of cells were used. Only SR1 could differentiate into multiple-lineage cell types under specific induction conditions. SR1 spheroids could differentiate to SR2 spheroids through epithelial-mesenchymal transition. Alkaline phosphatase (ALP) was highly expressed in SR1 spheroids, decreased in SR2 spheroids, and was absent in differentiated progenies in accordance with the loss of stemness properties. We verified that ALP can be a marker for ovarian CSLCs, and patients with greater ALP expression is related to advanced clinical stages and have a higher risk of recurrence and lower survival rate. The ALP inhibitor, levamisole, disrupted the self-renewal of ovarian CSLCs in vitro and tumor growth in vivo. In summary, this research provides a plastic ovarian cancer stem cell model and a new understanding of the cross-link between stem cells and cancers.This results show that ovarian CSLCs can be suppressed by levamisole. Our findings demonstrated that some ovarian CSLCs may restore ALP activity, and this suggests that inhibition of ALP activity may present a new opportunity for treatment of ovarian cancer. PMID:24280306

Liu, Kuei-Chun; Yo, Yi-Te; Huang, Rui-Lan; Wang, Yu-Chi; Liao, Yu-Ping; Huang, Tien-Shuo; Chao, Tai-Kuang; Lin, Chi-Kang; Weng, Shao-Ju; Ma, Kuo-Hsing; Chang, Cheng-Chang; Yu, Mu-Hsien; Lai, Hung-Cheng

2013-12-01

242

Strontium ranelate stimulates the activity of bone-specific alkaline phosphatase: interaction with Zn(2+) and Mg (2+).  

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Strontium ranelate (SR) is an orally administered and bone-targeting anti-osteoporotic agent that increases osteoblast-mediated bone formation while decreasing osteoclastic bone resorption, and thus reduces the risk of vertebral and femoral bone fractures in postmenopausal women with osteoporosis. Osteoblastic alkaline phosphatase (ALP) is a key enzyme involved in the process of bone formation and osteoid mineralization. In this study we investigated the direct effect of strontium (SR and SrCl2) on the activity of ALP obtained from UMR106 osteosarcoma cells, as well as its possible interactions with the divalent cations Zn(2+) and Mg(2+). In the presence of Mg(2+), both SR and SrCl2 (0.05-0.5 mM) significantly increased ALP activity (15-66 % above basal), and this was dose-dependent in the case of SR. The stimulatory effect of strontium disappeared in the absence of Mg(2+). The cofactor Zn(2+) also increased ALP activity (an effect that reached a plateau at 2 mM), and co-incubation of 2 mM Zn(2+) with 0.05-0.5 mM SR showed an additive effect on ALP activity stimulation. SR induced a dose-dependent decrease in the Km of ALP (and thus an increase in affinity for its substrate) with a maximal effect at 0.1 mM. Co-incubation with 2 mM Zn(2+) further decreased Km in all cases. These direct effects of SR on osteoblastic ALP activity could be indicating an alternative mechanism by which this compound may regulate bone matrix mineralization. PMID:24737106

Fernández, Juan Manuel; Molinuevo, Maria Silvina; McCarthy, Antonio Desmond; Cortizo, Ana Maria

2014-06-01

243

Leucine Aminopeptidase, ?-Glucosidase and Alkaline Phosphatase Activity Rates and Their Significance in Nutrient Cycles in Some Coastal Mediterranean Sites  

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In aquatic microbial ecology, knowledge of the processes involved in the turnover of organic matter is of utmost importance to understand ecosystem functioning. Microorganisms are major players in the cycling of nutrients (nitrogen, phosphorus) and carbon, thanks to their enzymatic activities (leucine aminopeptidase, LAP, alkaline phosphatase, AP, and ?-glucosidase, ?-GLU) on organic polymers (proteins, organic phosphates and polysaccharides, respectively). Estimates of the decomposition rates of organic polymers are performed using fluorogenic compounds, whose hydrolysis rate allow us to obtain information on the “potential” metabolic activity of the prokaryotic community. This paper refers the enzyme patterns measured during recent oceanographic cruises performed in some coastal Mediterranean sites, not yet fully investigated in terms of microbial biogeochemical processes. Mean enzyme activity rates ranged from 5.24 to 5558.1 nM/h, from 12.68 to 244.73 nM/h and from 0.006 to 9.51 nM/h for LAP, AP and ?-GLU, respectively. The highest LAP and AP activity rates were measured in the Gulf of Milazzo (Tyrrhenian Sea) and in the Straits of Messina, in association with the lowest bacterioplankton abundance; in contrast, the lowest ones were found in the northern Adriatic Sea. ?-GLU was more active in the Straits of Messina. Activity rates were analysed in relation to the main environmental variables. Along the northern Adriatic coastal side affected by the Po river, significant inverse relationships linked LAP and AP with salinity, pointing out that fluvial inputs provided organic substrates for microbial metabolism. Both in the Gulf of Manfredonia and in the Straits of Messina, LAP and AP levels were inversely related with the concentration of nitrate and inorganic phosphorus, respectively. In the Gulf of Milazzo, high cell-specific AP measured in spite of phosphorus availability suggested the role of this enzyme not only in phosphorus, but also in carbon release.

Caruso, Gabriella

2010-01-01

244

Leucine Aminopeptidase, ?-Glucosidase and Alkaline Phosphatase Activity Rates and Their Significance in Nutrient Cycles in Some Coastal Mediterranean Sites  

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Full Text Available In aquatic microbial ecology, knowledge of the processes involved in the turnover of organic matter is of utmost importance to understand ecosystem functioning. Microorganisms are major players in the cycling of nutrients (nitrogen, phosphorus and carbon, thanks to their enzymatic activities (leucine aminopeptidase, LAP, alkaline phosphatase, AP, and ?-glucosidase, ?-GLU on organic polymers (proteins, organic phosphates and polysaccharides, respectively. Estimates of the decomposition rates of organic polymers are performed using fluorogenic compounds, whose hydrolysis rate allow us to obtain information on the “potential” metabolic activity of the prokaryotic community. This paper refers the enzyme patterns measured during recent oceanographic cruises performed in some coastal Mediterranean sites, not yet fully investigated in terms of microbial biogeochemical processes. Mean enzyme activity rates ranged from 5.24 to 5558.1 nM/h, from 12.68 to 244.73 nM/h and from 0.006 to 9.51 nM/h for LAP, AP and ?-GLU, respectively. The highest LAP and AP activity rates were measured in the Gulf of Milazzo (Tyrrhenian Sea and in the Straits of Messina, in association with the lowest bacterioplankton abundance; in contrast, the lowest ones were found in the northern Adriatic Sea. ?-GLU was more active in the Straits of Messina. Activity rates were analysed in relation to the main environmental variables. Along the northern Adriatic coastal side affected by the Po river, significant inverse relationships linked LAP and AP with salinity, pointing out that fluvial inputs provided organic substrates for microbial metabolism. Both in the Gulf of Manfredonia and in the Straits of Messina, LAP and AP levels were inversely related with the concentration of nitrate and inorganic phosphorus, respectively. In the Gulf of Milazzo, high cell-specific AP measured in spite of phosphorus availability suggested the role of this enzyme not only in phosphorus, but also in carbon release.

Gabriella Caruso

2010-03-01

245

Immobilization of alkaline phosphatase on solid surface through self-assembled monolayer and by active-site protection.  

Science.gov (United States)

Retaining biological activity of a protein after immobilization is an important issue and many studies reported to enhance the activity of proteins after immobilization. We recently developed a new immobilization method of enzyme using active-site protection and minimization of the cross-links between enzyme and surface with a DNA polymerase as a model system. In this study, we extended the new method to an enzyme with a small mono-substrate using alkaline phosphatase (AP) as another model system. A condition to apply the new method is that masking agents, in this case its own substrate needs to stay at the active-site of the enzyme to be immobilized in order to protect the active-site during the harsh immobilization process. This could be achieved by removal of essential divalent ion, Zn2+ that is required for full enzyme activity of AP from the masking solution while active-site of AP was protected with p-nitrophenyl phosphate (pNPP). Approximately 40% of the solution-phase activity was acquired with active-site protected immobilized AP. In addition to protection active-site of AP, the number of immobilization links was kinetically controlled. When the mole fraction of the activated carboxyl group of the linker molecule in self-assembled monolayer (SAM) of 12-mercaptododecanoic acid and 6-mercapto-1-ethanol was varied, 10% of 12-mercaptododecanoic acid gave the maximum enzyme activity. Approximately 51% increase in enzyme activity of the active-site protected AP was observed compared to that of the unprotected group. It was shown that the concept of active-site protection and kinetic control of the number of covalent immobilization bonds can be extended to enzymes with small mono-substrates. It opens the possibility of further extension of the new methods of active-site protection and kinetic control of immobilization bond to important enzymes used in research and industrial fields. PMID:24738440

Gao, En-Feng; Kang, Kyung Lhi; Kim, Jeong Hee

2014-06-01

246

Comparative inhibition of human alkaline phosphatase and diamine oxidase by bromo-levamisole, cimetidine and various derivatives.  

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Analogues of bromo-levamisole and guanidine derivatives including cimetidine are examined in vitro in order to investigate their comparative inhibition, towards alkaline phosphatase (ALP) from human liver and diamine-oxidase (DAO) from human placenta. Bromo-levamisole, considered as a potent selective uncompetitive inhibitor of ALP (Ki, 2.8.10(-6) M at pH 10.5) is shown to be a noncompetitive inhibitor of DAO (Ki = 7.10(-4) M). According to the structure-inhibition relationship, the imidazole ring is important for ALP and DAO inhibition. The phenyl ring of bromo-levamisole is required for ALP inhibition but not for DAO inhibition, which is mediated mainly by aminoguanidine or guanidine groups. These results have allowed the selection of cimetidine, an H2-antagonist but also an immunomodulating compound, as inhibitor of these two enzymes. Cimetidine is an uncompetitive inhibitor of ALP (Ki = 3.2.10(-3) M at pH 10.5), and a good inhibitor of DAO (I50 = 3.8.10(-4) M). The Ki of ALP is commonly calculated at pH 10.5, but to study the role of the enzyme at the physiological pH, the inhibition has also been performed at pH 7.4. The Ki values are only slightly affected by this pH variation. So far several compounds, including levamisole, imidazole, theophylline and aminoguanidine are known to possess immunomodulating activities in vivo and/or in vitro and inhibit ALP and/or DAO. Therefore, it seems reasonable to assume that the inhibition of enzymes is involved in the immunomodulating effects of these drugs, when the ranges of active concentrations are similar for these properties. PMID:3143366

Metaye, T; Mettey, Y; Lehuede, J; Vierfond, J M; Lalegerie, P

1988-11-15

247

Combinations of nonlabeled, {sup 125}I-labeled, and anti-idiotypic antiplacental alkaline phosphatase monoclonal antibodies at experimental radioimmunotargeting  

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Purpose: Placental alkaline phosphatase (PLAP) is a membrane-bound oncofetal antigen that can be used for radioimmunotargeting. Preinjection of nonlabeled monoclonal anti-PLAP antibody (H7) and postinjection of monoclonal anti-idiotypic anti-PLAP antibody ({alpha}H7) were used in order to improve the localization efficacy of {sup 125}I-labeled H7. Material and Methods: A human cervix adenocarcinoma cell line (HeLa Hep 2) was inoculated subcutaneously in 24 nude mice. Repeated quantitative radioimmunoscintigraphic recordings were performed on 27 occasions in each of the 24 mice during the observation period which lasted for nearly 3 months. The tumor and nontumor doses were calculated according to the Medical International Radiation Dose Committee formula on the basis of the scintigraphic data. Results: All tumors were clearly visualized as early as one day after injection of {sup 125}I-labeled H7. The remaining radioactivity was exclusively located in the tumors at days 30-81. As much as 12-16% of the injected dose/g accumulated in the tumors during the first 2 days after injection, and remained stable at this high level for approximately 10 days in all investigated groups. Radioactivity in the whole body was rapidly eliminated during the same time period. The highest tumor/nontumor dose ratio was obtained after a single injection of {sup 125}I-labeled H7. Conclusion: Neither a preinjection of nonlabeled H7 nor a postinjection of {alpha}H7 nor a combination of both strategies resulted in improved tumor/nontumor dose ratios compared to a single injection of labeled H7. The monoclonal antibody H7 has a rapid and high uptake, combined with a prolonged retention time in the tumors. The kinetic properties of H7 are different form antibodies targeting intracellular tumor antigens. (orig.).

Rossi Norrlund, R.; Hietala, S.O.; Riklund Aahlstroem, K. [Univ. Hospital, Umeaa (Sweden). Dept. of Diagnostic Radiology; Holback, D.; Johansson, L. [Univ. Hospital, Umeaa (Sweden). Dept. of Radiation Physics

1997-11-01

248

Combinations of nonlabeled, 125I-labeled, and anti-idiotypic antiplacental alkaline phosphatase monoclonal antibodies at experimental radioimmunotargeting  

International Nuclear Information System (INIS)

Purpose: Placental alkaline phosphatase (PLAP) is a membrane-bound oncofetal antigen that can be used for radioimmunotargeting. Preinjection of nonlabeled monoclonal anti-PLAP antibody (H7) and postinjection of monoclonal anti-idiotypic anti-PLAP antibody (?H7) were used in order to improve the localization efficacy of 125I-labeled H7. Material and Methods: A human cervix adenocarcinoma cell line (HeLa Hep 2) was inoculated subcutaneously in 24 nude mice. Repeated quantitative radioimmunoscintigraphic recordings were performed on 27 occasions in each of the 24 mice during the observation period which lasted for nearly 3 months. The tumor and nontumor doses were calculated according to the Medical International Radiation Dose Committee formula on the basis of the scintigraphic data. Results: All tumors were clearly visualized as early as one day after injection of 125I-labeled H7. The remaining radioactivity was exclusively located in the tumors at days 30-81. As much as 12-16% of the injected dose/g accumulated in the tumors during the first 2 days after injection, and remained stable at this high level for approximately 10 days in all investigated groups. Radioactivity in the whole body was rapidly eliminated during the same time period. The highest tumor/nontumor dose ratio was obtained after a single injection of 125I-labeled H7. Conclusion: Neither a preinjection of nonlabeled H7 nor a postinjection of ?H7 nor a combination of both strategies resulted in improved tumor/nontumor dose ratios compared to a single injection of labeled H7. The monoclonal antibody H7 has a rapid and high uptake, combined with a prolonged retention time in the tumors. The kinetic properties of H7 are different form antibodies targeting intracellular tumor antigens. (orig.)

1997-11-01

249

High sequence variability, diverse subcellular localizations, and ecological implications of alkaline phosphatase in dinoflagellates and other eukaryotic phytoplankton  

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Full Text Available Alkaline phosphatase (AP is a key enzyme for phytoplankton to utilize dissolved organic phosphorus (DOP when dissolved inorganic phosphorus is limited. While three major types of AP and their correspondingly diverse subcellular localization have been recognized in bacteria, little is known about AP in eukaryotic phytoplankton such as dinoflagellates. Here, we isolated a full-length AP cDNA from a latest-diverging dinoflagellate genus Alexandrium, and conducted comparative analyses with homologs from a relatively basal (Amphidinium carterae and late-diverging (Karenia brevis lineage of dinoflagellates as well as other eukaryotic algae. New data and previous studies indicate that AP is common in dinoflagellates and most other major eukaryotic groups of phytoplankton. AP sequences are more variable than many other genes studied in dinoflagellates, and are divergent among different eukaryotic phytoplankton lineages. Sequence comparison to the other characterized APs suggests that dinoflagellates and some other eukaryotic phytoplankton possess the putative AP as phoA type, but some other eukaryotic phytoplankton seem to have other types. Phylogenetic analyses based on AP amino acid sequences indicated that the “red-type” eukaryotic lineages formed a monophyletic group, suggesting a common origin of their APs. As different amino acid sequences have been found to predictably determine different spatial distribution in the cells, which may facilitate access to different pools of DOP, existing computational models were adopted to predict the subcellular localizations of putative AP in the three dinoflagellates and other eukaryotic phytoplankton. Results showed different subcellular localizations of APs in different dinoflagellates and other lineages. The linkage between AP sequence divergence, subcellular localization and ecological niche differentiation requires rigorous experimental verification, and this study now provides a framework for such a future effort.

SenjieLin

2012-07-01

250

In vitro modulation of alkaline phosphatase activity of Saccharomyces cerevisiae grown in low or high phosphate medium  

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Full Text Available Our objective was to characterize the modulation of the activity of Saccharomyces cerevisiae alkaline phosphatases (ALPs by classic inhibitors of ALP activity, cholesterol and steroid hormones, in order to identify catalytic similarities between yeast and mammalian ALPs. S. cerevisiae expresses two ALPs, coded for by the PHO8 and PHO13 genes. The product of the PHO8 gene is repressible by Pi in the medium. ALP activity from yeast (grown in low or high phosphate medium homogenates was determined with p-nitrophenylphosphate as substrate, pH 10.4 (lPiALP or hPiALP, respectively. Activation of hPiALP was observed with 5 mM L-amino acids (L-homoarginine _ 186%, L-leucine _ 155% and L-phenylalanine - 168% and with 1 mM levamisole (122%; percentage values, in comparison to control, of recovered activity. EDTA (5 mM and vanadate (1 mM distinctly inhibited hPiALP (2 and 20%, respectively. L-homoarginine (5 mM had a lower activating effect on lPiALP (166% and was the strongest hPiALP activator. Corticosterone (5 mM inhibited hPiALP to 90%, but no effect was observed in low phosphate medium. Cholesterol, ß-estradiol and progesterone also had different effects on lPiALP and hPiALP. A concentration-dependent activation of lPiALP minus hPiALP was evident with all three compounds, most especially with ß-estradiol and cholesterol. These results do not allow us to identify similarities of the behavior of S. cerevisiae ALPs and any of the mammalian ALPs but allow us to raise the hypothesis of differential regulation of S. cerevisiae ALPs by L-homoarginine, ß-estradiol and cholesterol and of using these compounds to discriminate between S. cerevisiae lPiALP and hPiALP.

J Fernandes

2008-01-01

251

Red yeast rice stimulates osteoblast proliferation and increases alkaline phosphatase activity in MC3T3-E1 cells.  

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Red yeast (Monascus purpureus) is used as a traditional hypocholesterolemic dietary food component in Asia due to its bioactive component, lovastatin. Recently, new evidence suggesting that the statins in red yeast enhance bone formation has been reported, but more research is still needed in order to support these claims of osteogenic effects. Therefore, in this study, we hypothesized that red yeast rice (in which red yeast is fermented) can improve osteogenic function through osteoblast cell proliferation and differentiation. We studied the effect of methanol extract of red yeast rice powder (RYRP) on osteoblast proliferation and differentiation by measuring mitochondrial enzyme activity and bone marker alkaline phosphatase (ALP) activity, respectively. Osteoblast-like MC3T3-E1 cells were cultured in various concentrations of RYRP methanol extract (0.001-1 mg/mL) during the osteoblast differentiation period (1, 5, 10, and 15 days). As measured by 3-[4,5-dimethylthiazol-2-y]-2,5-diphenyltetrazolium bromide assay, RYRP extracts stimulated cell proliferation during a 24-hour period, compared to cooked white rice powder extract. The most pronounced effect was observed at the concentration range between 0.075 and 0.1 mg/mL. This RYRP stimulatory effect for cell proliferation was observed during the whole osteogenic period. Cellular (synthesized) ALP activity was increased at a RYRP extract concentration of 0.075 mg/mL during 15 days of culture, but the medium (secreted) ALP activity did not show any significant change. This cellular ALP activity stimulation by RYRP extract was confirmed by the staining of ALP activity on cell matrix layers for matrix calcification. The results imply that RYRP extract may increase osteogenic effect by stimulating cell proliferation and ALP activity in osteoblastic cells. PMID:20797483

Cho, Young-Eun; Alcantara, Ethel; Kumaran, Santhy; Son, Kun-Ho; Sohn, Ho-Yong; Lee, Jong-Hwa; Choi, Chung-Sig; Ha, Tae-Youl; Kwun, In-Sook

2010-07-01

252

Distinct expression of alkaline phosphatase activity in epilimnetic bacteria: Implication for persistent DOC consumption in a P-limited reservoir  

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In a P-deficient system, P availability usually controls the microbial activity and thus the ecosystem function. Thingstad et al. (1997) first addressed a 'Malfunctioning Microbial-loop' theory, which stated that low bacterial production (BP) caused by insufficient nutrient supply would result in DOC accumulation in an oligotrophic ecosystem. In this study we re-examined the theory by conducting seasonal patterns and correlations among soluble reactive phosphate (SRP) and DOC, microbial abundances (picocyanobacteria, bacteria, and heterotrophic nanoflagellate; HNF) and activities (primary production, bacterial production, and alkaline phosphatase activity; APA) coupled with enzyme-labeled fluorescence (ELF) assays on bacterioplankton in a subtropical reservoir sharing the common features, nitrate-replete and P-deficient, with most natural freshwater system during Oct 2007-Oct 2008. Persistently high APA was recorded during most of time, implying that the system was P-deficient. Size fractionated APA and ELF assay revealed that bacteria were the major APA contributor. However, significantly low epilimnion DOC was recorded during the stratified summer season accompanying with high BP and APA as well as high PP, implying that heterotrophic bacteria can well sustain in P-deficient system by utilizing DOP to rapidly lower down DOC under relatively high PP. Such findings oppose the 'Malfunctioning Microbial-loop' theory. On the other hand, strong epilimnetic DOC accumulation occurred in Oct 2007 under low light and low PP condition accompanying with high abundance of HNF, implying that HNF grazing may contribute to a certain degree of DOC accumulation. Correlation matrix supported our suggestions. This study testified the DOC dynamics in P-deficient ecosystem are tightly coupled with the source (PP and grazing) and sink (BP). We also suggested that in SRP-limited freshwater systems bacteria are capable of breaking down autochthonous DOC to reduce the chance of DOC accumulation during transport even dam construction continues to increase.

Tseng, Y.; Kao, S.; Shiah, F.

2013-12-01

253

Dissolved phosphorus pools and alkaline phosphatase activity in the euphotic zone of the western North Pacific Ocean.  

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Full Text Available We measured pools of dissolved phosphorus (P, including dissolved inorganic P (DIP, dissolved organic P (DOP and alkaline phosphatase (AP-hydrolyzable labile DOP (L-DOP, and kinetic parameters of AP activity (APA in the euphotic zone in the western North Pacific Ocean. Samples were collected from one coastal station in Sagami Bay, Japan, and three offshore stations between the North Pacific Subtropical Gyre (NPSG and the Kuroshio region. Although DIP concentrations in the euphotic zone at all stations were equally low, around the nominal method detection limit of 20 nmol L?1, chlorophyll a (Chl a concentrations were one order of magnitude greater at the coastal station. DOP was the dominant P pool, comprising 62–92% of total dissolved P at and above the Chl a maximum layer (CML. L-DOP represented 22–39% of the total DOP at the offshore stations, whereas it accounted for a much higher proportion (about 85% in the coastal surface layers. Significant correlations between maximum potential AP hydrolysis rates and DIP concentrations or bacterial cell abundance in the offshore euphotic zone suggest that major APA in the oligotrophic surface ocean is from bacterial activity and regulated largely by DIP availability. Although the range of maximum potential APA was comparable among the environmental conditions, the in situ hydrolysis rate of L-DOP in the coastal station was 10 times those in the offshore stations. L-DOP turnover time at the CML ranged from 4.5 d at the coastal station to 84.4 d in the NPSG. The ratio of the APA half saturation constant to the ambient L-DOP concentration decreased markedly from the NPSG to the coastal station. There were substantial differences in the rate end efficiency of DOP remineralization and its contribution as the potential P source between the low-phosphate/high biomass coastal ecosystem and the low-phosphate/low biomass oligotrophic ocean.

MasahiroSuzumura

2012-03-01

254

Utilidad de la fosfatasa alcalina urinaria como marcador precoz de lesión tubular renal / Utility of urinary alkaline phosphatase as early marker of renal tubular failure  

Scientific Electronic Library Online (English)

Full Text Available SciELO Argentina | Language: Spanish Abstract in spanish El objetivo de este estudio fue determinar la actividad de la fosfatasa alcalina urinaria (FALur) para evaluar precozmente lesión tubular y su utilidad diagnóstica. Los pacientes estudiados fueron: 20 Controles (C), 11 hipertensos (HTA), 23 diabéticos (DBT) y 34 con insuficiencia renal de diverso or [...] igen (IRDO). Se realizaron las determinaciones de: creatinina, clearence de creatinina (Jaffé cinético), FAL sérica y urinaria (cinético DGKC), microalbuminuria (inmunoturbidimétrico), proteiunuria (turbidimétrico), uroproteinograma, SDS-PAGE (al 12,5%) e isoenzimograma de FAL. La FAL sérica hallada fue normal, sin diferencia entre grupos y sin relación con el aumento de la FALur. El valor de corte recomendado para FALur fue de 8 UI/L. La FALur estuvo elevada en HTA e IRDO y normal en individuos con DBT. Los aumentos de FALur en IRDO se relacionaron con la lesión tubular estructural y en pacientes con HTA podrían relacionarse con alteración tubular precoz. Se propone la determinación de FALur para la detección temprana de lesión tubular ante falla renal establecida o en individuos con riesgo de desarrollarla, y se establece su utilidad en pacientes: - con DBT y HTA para seguimiento (junto a microalbuminuria y clearence de creatinina), - internados en riesgo de insuficiencia renal aguda: para orientar tratamientos, - con insuficiencia renal crónica: como indicador de lesión y pronóstico. Abstract in english The objective of this study was to determine the activity of urinary Alkaline Phosphatase (ALPur) to evaluate early tubular failure and its diagnostic usefulness. The patients studied were: 20 Controls (C), 11 with Hipertensión (HTA), 23 Diabetic (DBT) and 34 with renal Insufficiency of diverse orig [...] in (IRDO). The creatinine, creatinine clearence (kinetic Jaffé) serum and urinary ALP (kinetic DGKC), microalbuminuria (Immunoturbidimetric), proteiunuria (Turbidimetric), uroproteinogram, SDS-PAGE (12.5%) and ALP isoenzymes determinations were made. The results indicate that serum ALP was normal, without difference between groups, and no relation with the increase in ALPur. Recommended cut-off value of ALPur was 8 UI/L. ALPur was elevated in HTA and IRDO, and normal in DBT. Increases in ALPur in IRDO were related to the structural tubular injury, and those in HTA could be related to early tubular alteration. Determination of ALPur is proposed for early detection of tubular injury, before renal failure is established or when there is risk of developing it, establishing its usefulness in: - DBT and HTA patients: screening (together with microalbuminuria and creatinine clearence). - Hospitalized patients in risk of acute renal insufficiency: in order to orient treatments. - Patients with chronic renal insufficiency: as an indicator of injury and prognosis.

Di Carlo, María Beatriz; Gomez, Alejandra Gabriela; Madalena, Leticia Bibiana; Facio, María Laura; Pizzolato, Marco Antonio; Negri, Gustavo Alberto.

255

Serum levels of bone alkaline phosphatase in breast and prostate cancers with bone metastasis  

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BALP activity in the sera of metastatic patients of breast and prostate malignancy has increased significantly. Our studies with patients in India conform the earlier reports that BALP may have a useful complementary role in the early diagnosis of bone metastases.

Ramaswamy, Girija; Rao, Vasanti R.; Krishnamoorthy, Lakshmi; Ramesh, G.; Gomathy, R.; Renukadevi, D.

2000-01-01

256

In vitro modulation of alkaline phosphatase activity of Saccharomyces cerevisiae grown in low or high phosphate medium  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Our objective was to characterize the modulation of the activity of Saccharomyces cerevisiae alkaline phosphatases (ALPs) by classic inhibitors of ALP activity, cholesterol and steroid hormones, in order to identify catalytic similarities between yeast and mammalian ALPs. S. cerevisiae expresses two [...] ALPs, coded for by the PHO8 and PHO13 genes. The product of the PHO8 gene is repressible by Pi in the medium. ALP activity from yeast (grown in low or high phosphate medium) homogenates was determined with p-nitrophenylphosphate as substrate, pH 10.4 (lPiALP or hPiALP, respectively). Activation of hPiALP was observed with 5 mM L-amino acids (L-homoarginine _ 186%, L-leucine _ 155% and L-phenylalanine - 168%) and with 1 mM levamisole (122%; percentage values, in comparison to control, of recovered activity). EDTA (5 mM) and vanadate (1 mM) distinctly inhibited hPiALP (2 and 20%, respectively). L-homoarginine (5 mM) had a lower activating effect on lPiALP (166%) and was the strongest hPiALP activator. Corticosterone (5 mM) inhibited hPiALP to 90%, but no effect was observed in low phosphate medium. Cholesterol, ß-estradiol and progesterone also had different effects on lPiALP and hPiALP. A concentration-dependent activation of lPiALP minus hPiALP was evident with all three compounds, most especially with ß-estradiol and cholesterol. These results do not allow us to identify similarities of the behavior of S. cerevisiae ALPs and any of the mammalian ALPs but allow us to raise the hypothesis of differential regulation of S. cerevisiae ALPs by L-homoarginine, ß-estradiol and cholesterol and of using these compounds to discriminate between S. cerevisiae lPiALP and hPiALP.

J, Fernandes; R, Amorim; I, Azevedo; M.J, Martins.

257

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

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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±2.5>EMD/BG(64.78±3.04>EMD/HG(55.40±3.89?EMD/BM(54.75±4.17>BG (51.32±2.76>HG(49.92±2.4>BM(48.14±1.4>Control(46.29±1.39>EMD/CP (37.46±3.54>CP(32.12±1.49 Conclusion: There was no additive effect on osteoblast adhesion/ALP activity following exposure to an EMD/polymer combination. EMD/collagen positively influences osteoblast differentiation in a time dependent manner.

Thangakumaran S

2009-01-01

258

An evaluation on the activity level of Aspartate aminotransferase and Alkaline phosphatase nzymes in peri-implant sulcus fluid  

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Full Text Available Statement of Problem: The correlation between the activity of aspartate aminotransferase (AST and alkaline phosphatase (ALP enzymes in gingival sulcular fluid (GCF with inflammation and periodontal attachment loss has been proved, however there are not adequate studies about dental implants. Purpose: The aim of present study was to investigate the presence and activity level of AST & ALP and their correlation with pocket depth (PD and bleeding of peri-implant slcular fluid (PISF, and to evaluate the possibility of using these assessments as a diagnostic index in oral implantology. Material and Methods: In this study, 41 implants as test group and 41 contralateral teeth as control group, in 21 patients were evaluated. At first visit, the general information about implants and the values of pocket probing depth (PPD, modified sulcus bleeding index (mSBl and modified plaque index (mPI were recorded. At the second visit, samples of GCF/PISF were collected. AST & ALP activity was determined spectrophotometrically and data were analyzed by "t", "Mann-Whitney" tests and Pearson Spearman correlation coefficient."nResults: The results showed that there was a significant difference in the activity of AST between two study groups (P<0.0001. The average activity of ALP in test group was more than control group but the difference was not significant. After elimination of the confounding variables, the average AST in test group was 54.6 (S£=2.3 and in control groups was 44.8 (SE=2.3 (P=0.004. The average ALP in test group (SE=2.2 and in control (SE=2.2 were 36.6 and 35.4, respectively. Values of AST and ALP were positively correlated with other clinical parameters such as PD and mSBI which was significant in test group."nConclusion: The present study suggests that PISF analysis could be considered as a proper diagnostic strategy in the evaluation of dental implant success.

Paknegad M. Assistant Professor

2003-07-01

259

Kinetics and crystal structure of a mutant Escherichia coli alkaline phosphatase (Asp-369-->Asn): a mechanism involving one zinc per active site.  

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Using site-directed mutagenesis, an aspartate side chain involved in binding metal ions in the active site of Escherichia coli alkaline phosphatase (Asp-369) was replaced, alternately, by asparagine (D369N) and by alanine (D369A). The purified mutant enzymes showed reduced turnover rates (kcat) and increased Michaelis constants (Km). The kcat for the D369A enzyme was 5,000-fold lower than the value for the wild-type enzyme. The D369N enzyme required Zn2+ in millimolar concentrations to become...

Tibbitts, T. T.; Xu, X.; Kantrowitz, E. R.

1994-01-01

260

Production of placental alkaline phosphatase (PLAP) and PLAP-like material by epithelial germ cell and non-germ cell tumours in vitro.  

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Placental and placental-like alkaline phosphatase (PLAP) levels in the culture media of 87 cell lines of neoplastic and 'normal' origin were measured by a conventional immunosorbent enzymatic assay (IAEA) and by a new immunoradiometric assay (IRMA). The IRMA detected immunoreactive PLAP in 37 of 80 (46%) human epithelial and germ cell cultures, while the IAEA detected PLAP in only 25 (33%). Of the 52 non-germ cell tumour cultures, the IRMA detected expression in 24 (46%) and the IAEA in only ...

1994-01-01

 
 
 
 
261

Oxovanadium (iv) complexes with n/o- and o-donor ligands: their synthesis, characterization, semiempirical study and alkaline phosphatase activity (abstract)  

International Nuclear Information System (INIS)

Various N/O- and O-donor ligands and their oxovanadium complexes have been synthesized and characterized by different techniques such as FTIR, elemental analysis, thermogravimetery and conductometry. The IR data show the bidentate nature of the ligands and reveals hexa-coordinated geometry in the solid state which is also confirmed by semi-empirical study. Conductance measurements reveal the non-electrolytic nature of the complexes. These complexes have been checked for their alkaline phosphatase activity in the presence and absence of inhibitor which shows that by the addition of inhibitor the activity of enzyme decreases and at higher concentration it is completely inhibited. (author)

2011-01-01

262

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

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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 measurement of 25-hydroxy vitamin-D is recommended in epileptic children taking anti-epileptic dugs. Supplemental vitamin-D administration in such patients may be helpful.

Keyhani doost Z

2011-01-01

263

Effect of Zinc from Zinc Sulfate on Ewes` Weight, Milk Yield, Zn Concentrations in Serum and Serum Alkaline Phosphates Activity of Varamini Ewes  

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This experiment was conducted to investigate the effect of feeding supplemental zinc (zinc sulfate) in different levels (0, 15 and 30 mg/kg) on ewes weight, milk production, Zn concentrations in serum and serum alkaline phosphates activity. Thirty lactating Varaminni ewes were assigned to three experimental groups according to their live body weights, milk production and lambs sex in a completely randomized design. Ewes were fed a basal diet containing alfalfa, wheat straw, cottonseed meal, b...

2008-01-01

264

The expression of alkaline phosphatase, osteopontin, osteocalcin, and chondroitin sulfate during pectoral fin regeneration in Carassius auratus gibelio: a combined histochemical and immunohistochemical study.  

Science.gov (United States)

Dermal bone is an important component of the teleost fins, and its ability to regenerate after fin amputation appears to be unlimited. The organic bone matrix contain type I collagen fibers, proteoglycans enriched in chondroitin sulfate, and noncollagenous matrix protein such as osteocalcin, osteopontin, and osteonectin. These molecules are synthesized by fin osteoblasts. Inorganic components chiefly consist of calcium and phosphate that form crystals of hydroxyapatite. Fin rays are described as models to study ossification. Due to this, the identification of the components involved in the synthesis of the organic and inorganic components of lepidotrichial bone are of great interest for the analysis of skeletal disorders in fish ossification. The present study investigates expression of alkaline phosphatase, osteopontin, osteocalcin, and chondroitin sulfate during pectoral fin regeneration in Carassius auratus gibelio. Alkaline phosphatase reaction has been found in the epidermis covering the wound, proximal blastema, near the cells that surround newly-formed lepidotrichia matrix and the tips of regenerating fin rays. Osteopontin has been observed throughout the regeneration blastema but excluded from the scleroblasts lining the inner side of the lepidotrichia. Osteocalcin and chondroitin sulfate expression coincides with the onset of mineralization of lepidotrichial matrix, suggesting its involvement in bone mineralization. PMID:23302437

Stavri, Simona; Zarnescu, Otilia

2013-02-01

265

Assay Format as a Critical Success Factor for Identification of Novel Inhibitor Chemotypes of Tissue-Nonspecific Alkaline Phosphatase from High-Throughput Screening  

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Full Text Available The tissue-nonspecific alkaline phosphatase (TNAP isozyme is centrally involved in the control of normal skeletal mineralization and pathophysiological abnormalities that lead to disease states such as hypophosphatasia, osteoarthritis, ankylosis and vascular calcification. TNAP acts in concert with the nucleoside triphosphate pyrophosphohydrolase-1 (NPP1 and the Ankylosis protein to regulate the extracellular concentrations of inorganic pyrophosphate (PPi, a potent inhibitor of mineralization. In this review we describe the serial development of two miniaturized high-throughput screens (HTS for TNAP inhibitors that differ in both signal generation and detection formats, but more critically in the concentrations of a terminal alcohol acceptor used. These assay improvements allowed the rescue of the initially unsuccessful screening campaign against a large small molecule chemical library, but moreover enabled the discovery of several unique classes of molecules with distinct mechanisms of action and selectivity against the related placental (PLAP and intestinal (IAP alkaline phosphatase isozymes. This illustrates the underappreciated impact of the underlying fundamental assay configuration on screening success, beyond mere signal generation and detection formats.

José Luis Millán

2010-04-01

266

The lipid raft-bound alkaline phosphatase activity increases and the level of transcripts remains unaffected in liver of merosin-deficient LAMA2dy mouse.  

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Alkaline phosphatase (AP) and other proteins add glycosylphosphatidylinositol (GPI) before addressing to raft domains of the cell membrane. Our previous report showing an increased density of lipid rafts in muscle of dystrophic Lama2dy mice prompted us to compare livers of normal (NL) and dystrophic mice (DL) for their levels of rafts. With this aim, hepatic rafts were isolated as Triton X-100 resistant membranes, and identified by their abundance of flotillin-2, alkaline phosphatase (AP) and other raft markers. The comparable abundance of cholesterol and flotillin-2 in rafts of NL and DL contrasted with the double AP activity both in rafts of DL and whole DL. The AP mRNA level was the same in NL and DL. Sedimentation analysis profiles revealed AP activity of NL distributed between dimeric (dAP) and monomeric AP (mAP), whose proportions and lectin-binding extent changed in DL. The increased AP activity and changed AP glycosylation in DL, the prevalence of mAP in NL and the enhanced stability of dAP in DL demonstrated the critical role that glycosylation and oligomerization play for AP catalysis. The higher AP activity of DL probably arises from dystrophy-associated changes in glycosyl transferases, which alter AP glycosylation and subunit folding with profitable effects for AP stability and catalysis. PMID:24680793

Montenegro, María Fernanda; Moral-Naranjo, María Teresa; Campoy, Francisco J; Muñoz-Delgado, Encarnación; Vidal, Cecilio J

2014-06-01

267

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

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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.

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: dtakai-ind@umin.ac.jp [Department of Clinical Laboratory, The University of Tokyo Hospital, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8655 (Japan)

2010-08-27

268

Alkaline phosphatase activity and its relationship to inorganic phosphorus in the transition zone of the North-western African upwelling system  

Science.gov (United States)

The enzymatic activity of alkaline phosphatase (APA) was studied in the transition zone between the African upwelling system and the open ocean waters of the Canary Islands region. This region is recurrently dominated by the presence of upwelling filaments that may transport nutrient-enriched waters out into the open ocean before nutrients become exhausted by plankton. Turnover rates by APA were generally low in the whole region, but detectable in all the measurements carried out. On average, turnover rates were higher in the upwelling stations, and APA in those waters seemed to be mainly generated by heterotrophic bacteria to supply easily assimilable organic C. APA outside the upwelling area showed an inverse hyperbolic relationship with increasing phosphate, suggesting the presence of both constitutive and Pi-inducible APA. In these offshore waters, a threshold of 0.1 ?M of phosphate could be defined for the regulatory function of Pi on APA. Thus, APA in nutrient-poor waters seemed to be induced to compensate for Pi-deficiency. Turnover rates in the filaments showed basal (probably constitutive) levels, whereas they increased in the surrounding waters, where phosphate concentration presumably did not satisfy plankton P-demands. The fertilising effect of the filaments and associated cyclonic eddies extended to at least 175 km offshore, where basal alkaline phosphatase activities were still found. The magnitude of this effect depends probably on the intensity of upwelling events and the degree of recirculation of filament water back to the coastal jet.

Sebastián, Marta; Arístegui, Javier; Montero, María F.; Escanez, Jose; Xavier Niell, F.

2004-08-01

269

Serum Chemistries of 'Coturnix coturnix japonica' Given Dietary Manganese Oxide (Mn3O4).  

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Plasma creatinine and inorganic phosphorus were increased in manganese oxide (Mn3O4)-treated adult male Coturnix quail, but BUN, BUN/creatinine ratio, uric acid, and total calcium were decreased. Serum enzymes (alkaline phosphatase, glutamic oxaloacetic t...

F. W. Edens J. W. Laskey

1990-01-01

270

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

International Nuclear Information System (INIS)

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 capacity to the components from SCWPs of F. verticillioides, and enzyme-linked immunosorbent assays revealed that the detection limit of the fungus was below 10?2 ?g mL?1, superior to the scFv antibody. The fusion protein was able to detect fungal concentrations as low as 10?3 mg g?1 of maize grains in both naturally and artificially contaminated samples. Thus, the fusion can be applied in rapid and simple diagnosis of Fusarium contamination in field and stored grain or in food

2013-02-18

271

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

Energy Technology Data Exchange (ETDEWEB)

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 capacity to the components from SCWPs of F. verticillioides, and enzyme-linked immunosorbent assays revealed that the detection limit of the fungus was below 10{sup ?2} ?g mL{sup ?1}, superior to the scFv antibody. The fusion protein was able to detect fungal concentrations as low as 10{sup ?3} mg g{sup ?1} of maize grains in both naturally and artificially contaminated samples. Thus, the fusion can be applied in rapid and simple diagnosis of Fusarium contamination in field and stored grain or in food.

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: ycliao06@yahoo.com.cn [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)

2013-02-18

272

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

Science.gov (United States)

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. PMID:22743140

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

2013-03-01

273

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

International Nuclear Information System (INIS)

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 ?g mm-2) yielded the same electrodic improvements but with better analytical properties

2008-05-12

274

Copper(II) complexes of methimazole, an anti Grave's disease drug. Synthesis, characterization and its potential biological behavior as alkaline phosphatase inhibitor.  

Science.gov (United States)

Methimazole (MeimzH) is an anti-thyroid drug and the first choice for patients with Grave's disease. Two new copper(II) complexes of this drug: [Cu(MeimzH)(2)(NO(3))(2)]*0.5H(2)O and [Cu(MeimzH)(2)(H(2)O)(2)](NO(3))(2)*H(2)O were synthesized and characterized by elemental analysis, dissolution behavior, thermogravimetric analysis and UV-vis, diffuse reflectance, FTIR and EPR spectroscopies. As it is known that copper(II) cation can act as an inhibitor of alkaline phosphatase (ALP), the inhibitory effect of methimazole and its copper(II) complexes on ALP activity has also been investigated. PMID:20012553

Urquiza, Nora M; Manca, Silvia G; Moyano, María A; Dellmans, Raquel Arrieta; Lezama, Luis; Rojo, Teófilo; Naso, Luciana G; Williams, Patricia A M; Ferrer, Evelina G

2010-04-01

275

Validation of an analytical procedure for the determination of alkaline and alkaline- earth metals in mices´serum  

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Full Text Available The interaction between drugs or toxic substances with human body, and in particular those which transit natural barriers and get inside blood vessels, could to produce incorrect ions balance due to formation of aducts or for biological membrane damage and chemical alteration of ions in the surrounding. For this reason it is important to determine those elements in serum of test animals to establish experimental conditions for preclinic toxicological tests. In the present work, the results of design and validation of analytical procedures for the determination of sodium, potassium, calcium and magnesium from NMRI mouse’s serum using flame atomic absorption spectrometry. The procedures are recommended because they use a very little amount of serum sample, as well as their performance characteristics, like precision, trueness, sensibility and selectivity.

Alvarez M

2008-04-01

276

Pretreatment of Rats with ?-tocopherol Alter Liver and Kidney Protein, Alkaline Phosphatase Activity and Phospholipid Profile after 24 Hour Intoxication with Cadmium  

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Full Text Available Cell membrane composition and fluidity are altered in diseases and previous reports suggest that membrane lipid is altered in heavy metal toxicity. This study was carried out to assess the effect of cadmium alone and its combination with different doses (75, 150 and 750 mg of ?-tocophenylacetate on the phospholipid profile and alkaline phosphatase activity in the kidney and liver of rats. The results obtained show that in these tissues, cadmium significantly (p<0.05 increased the levels of protein compared with the control, in the liver it significantly raised total lipid levels and decreased it in the kidney. Vitamin E in the form of ?-tocophenylacetate reversed these observations. Cadmium decreased alkaline phosphatase activity significantly (p<0.05 in both tissues; a trend that was counteracted by ?-tocophenylacetate supplementation in a dose dependent pattern. In both the liver and kidney, cadmium treatment reduced the levels of phosphatidylcholine (PC, phosphatidylethanolamine (PE and increased sphingomyelin (SGM. It raised the PC/PE and SGM/PC ratios, index of membrane fluidity. Vitamin E supplementation significantly reduced the SGM/PC ratios in a manner that appears to be dose related. In the liver the effect of the vitamin on the SGM/PC ratio range from about 400 to 300 to 800% in the 75, 150 and 750 mg supplemented groups, respectively. A similar trend was also observed in the kidney. We hypothesize that decreased membrane fluidity occasioned by increase in SGM/PC ratio occur in early cadmium toxicity and that vitamin E cushions the effect of cadmium by decreasing SGM/PC ratio.

G.E. Eriyamremu

2006-01-01

277

Influence of a Nigerian-like Diet on Calcium, Phosphate and Alkaline Phosphatase Levels in the Plasma and Bone of Cadmium Exposed Rats  

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Full Text Available The purpose of this study was to assess the role of a wholly compounded Nigerian diet on some indices of bone metabolism in oral cadmium toxicity. Nine weeks old Wistar albino rats were exposed to 100 ppm cadmium in their drinking water and fed a Nigerian-like diet (high in carbohydrate and fibre but low in protein for sixteen weeks. The study reveals that the Nigerian-like Diet (NLD caused less accumulation of cadmium in both the epiphysis and metaphysis of femur bones compared with the control diet. Plasma cadmium and phosphate levels did not change significantly with diet and cadmium exposure. Plasma alkaline phosphate activity was enhanced in rats fed with the NLD and cadmium exposure reduced the enzyme activity. The exposure to cadmium compared with the control significantly (p<0.05 reduced calcium and phosphate in both the epiphysis and metaphysis of the femur of rats, which was further enhanced by the NLD. The NLD increased alkaline phosphatase activity in both sections of the bone studied, though cadmium reduced the enzyme activity. The study shows that the NLD predispose rats to the debilitating effect of cadmium on bones by improving calcium and phosphate loss which promote bone resorption.

Samuel Ogheneovo Asagba

2006-01-01

278

Correlation of Serum Magnesium with Serum Parathormone Levels in Patients on Regular Hemodialysis  

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Secondary hyperparathyroidism (SHPT) is a common, important, and treatable complication of end-stage renal disease. This study was conducted to investigate the role of serum magnesium (Mg) in regulating the secretion of parathyroid hormone (PTH) by the parathyroid gland in patients on maintenance hemodialysis (HD). Pre-dialysis serum levels of calcium (Ca), phosphorus (P), Mg, alkaline phosphatase (ALP), intact serum PTH (iPTH), serum 25-hydroxy Vitamin D (25-OH Vit D) and plasma bicarbonate ...

Baradaran Azar; Nasri Hamid

2006-01-01

279

Estado nutricional, consumo de lácteos y niveles séricos de calcio, fósforo y fosfatasas alcalinas en escolares de Mérida / Nutritional status, consumption of dairy products and levels sericos of calcium, phosphorus, and alkaline phosphatases in schoolchildren of Mérida  

Scientific Electronic Library Online (English)

Full Text Available SciELO Venezuela | Language: Spanish Abstract in spanish Se realizó una investigación de Campo de Tipo Descriptiva Correlacional y de corte transversal para determinar el estado nutricional, consumo de lácteos y niveles séricos de calcio, fósforo, y fosfatasas alcalinas en escolares del 1er, 3er y 5to grado de la U.E "Rafael Antonio González" de la comuni [...] dad de Mesa Bolívar en el año 2007. La población estuvo conformada por la matricula escolar de 171 estudiantes. Se determinó la muestra con el método estratificado aleatorio simple, obteniéndose 47% de la matricula escolar, correspondiendo 80 niños distribuidos por grado: 21 niños en 1ero, 28 en 3ero y 31 en 5to, en edades comprendidas entre 6 a 12 años. Se determinó la cantidad y la frecuencia de consumo de productos lácteos para lo cual, se diseñó un cuestionario "ad hoc" contentivo de 10 ítems relacionados con la frecuencia de consumo, cantidad y tipo de lácteos. Se realizó evaluación nutricional a través de la Combinación de Indicadores (Peso para la Talla y Talla para la Edad) utilizando las tablas de Evaluación de la Organización Mundial de la Salud. Se determinaron los valores séricos de calcio, fósforo y fosfatasas alcalinas. Los escolares presentan 32,6% de malnutrición; tanto los niños (6-10 años y 11-12 años) como las niñas (8-12 años) presentaron un porcentaje de adecuación diario de calcio bajo (77,16%, 28,57% y 38,96%) respectivamente y 60% tienen hipocalcemia. Existe significancia estadística entre los niveles séricos de calcio y fósforo con el consumo diario promedio de calcio (p 0,05 y p 0,04). No hubo relación estadísticamente significativa entre el consumo de productos lácteos y el estado nutricional de los escolares. El estado nutricional de los escolares no depende del consumo diario de productos lácteos, sin embargo, dicho consumo si afecta los niveles séricos de calcio y fósforo. Abstract in english A cross-sectional descriptive correlational field research was conducted in order to determine the nutritional status, consumption of milk and serum levels of calcium, phosphorus, and alkaline phosphatase in students of 1st, 3rd and 5th grades of the "Rafael Antonio Gonzalez "school in Mesa Bolívar [...] in 2007. The population consisted of 171 students. We determined the sample with a simple random stratified method, yielding 47% of school enrollment, corresponding to 80 children distributed by grade: 21 children in 1st, 28 in 3rd, 31 in 5th, aged 6 to 12 years old. The amount and frequency of consumption of dairy products, with an "ad hoc" questionnaire designed containing 10 items related to the frequency of consumption, quantity and type of dairy product. Nutritional assessment was carried out by means of the combination of indicators (weight for height and height for age) using the tables of evaluation of the World Health Organization. Values were determined in serum calcium, phosphorus and alkaline phosphatase. The students had 32,6% of malnutrition, both boys (6-10 years and 11-12 years) and girls (8-12 years) had an adequate percentage of low calcium daily intake(77.16%, 28. 57% and 38.96%, respectively) and 60% had hypocalcemia. There is statistical significance between serum calcium and phosphorus with an average daily intake of calcium (p 0.05 and p 0.04). There was no statistically significant relationship between dairy products consumption and nutritional status of schoolchildren. The nutritional status of schoolchildren does not depend on daily consumption of dairy products, however, that consumption does affect serum calcium and phosphorus.

Rojas, Lizbeth; Bastardo, Gladys; Sanz, Belquis; Da Silva, G. Beatriz; Quintero de Rivas, Yurimay; Angarita, Coromoto; Prada Briceño, Maribel.

280

Tartrate-resistant acid phosphatase isoform 5b: a novel serum marker for monitoring bone disease in multiple myeloma.  

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Tartrate-resistant acid phosphatase isoform-5b (TRACP-5b), a new marker reflecting osteoclast activity, and osteoprotegerin (OPG) were measured in 121 patients with multiple myeloma (MM) at diagnosis, and in 63 of them during pamidronate administration, to define their correlation with the extent of bone disease and disease activity in MM. Radiographic evaluation of the skeleton, measurement of other markers of bone remodelling, including N-terminal cross-linking telopeptide of type-I collagen (NTX), bone alkaline phosphatase and osteocalcin and of markers of disease activity (beta2-microglobulin, paraprotein, interleukin-6 (IL-6), were also performed. Levels of TRACP-5b were increased (p <.0001), while OPG was decreased in MM patients compared to controls (p <.01). TRACP-5b levels were associated with the radiographically assessed severity of bone disease (p <.0001) as well as with levels of NTX, IL-6 and beta2-microglobulin (p <.001, for each biochemical parameter, respectively). The combination of pamidronate with VAD-chemotherapy produced a reduction in TRACP-5b, NTX, IL-6, paraprotein and beta2-microglobulin levels from the 2nd month of treatment, with no effect on bone formation and OPG. A strong correlation was observed between changes in TRACP-5b and changes in NTX, IL-6 and beta2-microglobulin, while TRACP-5b predicted the disease progression in 5 patients. These findings suggest that TRACP-5b is increased in MM, reflects the extent of myeloma bone disease and may have a predictive value. TRACP-5b has also proved to be very useful for monitoring antimyeloma treatment, which had no effect on OPG levels. PMID:12845688

Terpos, Evangelos; de la Fuente, Josu; Szydlo, Richard; Hatjiharissi, Evdoxia; Viniou, Nora; Meletis, John; Yataganas, Xenophon; Goldman, John M; Rahemtulla, Amin

2003-09-01

 
 
 
 
281

Stimulation by parathyroid hormone of sup 45 Ca sup 2+ uptake in osteoblast-like cells: Possible involvement of alkaline phosphatase  

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We have investigated the actions of human PTH (hPTH-(1-34)) on the association of 45Ca2+ with two human (SaOS-2 and MG-63) and two rat (ROS 17/2.8 and UMR-106) osteoblast-like cell types. In SaOS-2 cells, hPTH-(1-34) binds to specific membrane receptors to activate adenylate cyclase. Treatment of SaOS-2 cells with hPTH-(1-34) resulted in an increase in 45Ca2+ uptake, in a dose-dependent fashion, up to 2- to 4-fold above control values. The increase was first evident at 10 min and persisted for at least 30 min. Treatment with nimodipine, a calcium channel antagonist, was without effect on the stimulatory action of PTH. A similar enhancement of cell-associated 45Ca2+ was observed when the cells were incubated with vasoactive intestinal peptide, which acts via different receptors to activate adenylate cyclase in SaOS-2 cells. Treatment with (Bu)2cAMP also induced an increase in cell-associated 45Ca2+. Pretreatment of SaOS-2 cells with hPTH-(1-34) for 4 h, which induced homologous desensitization to a second challenge with the same peptide for stimulation of cAMP production, did not attenuate the further enhancement of cell-associated 45Ca2+ by a second treatment with hPTH-(1-34). We then examined a possible relationship between alkaline phosphatase (ALPase) and 45Ca2+ uptake. SaOS-2 cells contained high levels of alkaline phosphatase activity and continuously released the enzyme into the medium. Release was enhanced by treatment with hPTH-(1-34) for 10 min. Incubation of cells with levamisole (an inhibitor of the liver/bone/kidney type of ALPase) resulted in a rapid decrease in basal and PTH-stimulated 45Ca2+ uptake, while treatment with L-Phe-Gly-Gly was without effect. Treatment of the cells with ALPase (bovine kidney) enhanced 45Ca2+ uptake. In MG-63 cells, a stimulatory effect of hPTH-(1-34) on cell-associated 45Ca2+ was also observed; however, hPTH-(1-34) did not stimulate cAMP production in MG-63 cells.

Fukayama, S.; Tashjian, A.H. Jr. (Harvard School of Public Health, Boston, MA (USA))

1990-04-01

282

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

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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. Conclusion The results demonstrate the biochemical modulation of scFv binding. Also, the scFvs bound to the active site and denied the access to the substrate. The selection strategy could generate specific anti-enzyme antibodies to PLAP that can potentially be used for targeting, for modulating enzyme activity in in vitro and in vivo and as probes for the active site. This strategy also has a general application in selecting antibodies from combinatorial libraries to closely related molecules and conformations.

Kala Mrinalini

2005-12-01

283

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

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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.

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

1977-12-22

284

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

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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 transitions occur. The first alkaline transition takes place at pH between 11.3 and 11.8, co-operatively dissociating at least six protons. The second alkaline transition takes place at pH between 11.8 and 12.0. It probably implies a reversible unfolding of the albumin molecule, increasing the distance between tryptophan-214 and bilirubin, and partly exposing the liganded bilirubin to the solvent.

Honore, B; Frandsen, P C

1986-01-01

285

One-Step Detection of Aflatoxin-B(1) Using scFv-Alkaline Phosphatase-Fusion Selected from Human Phage Display Antibody Library  

DEFF Research Database (Denmark)

A unique human phage display library was used to successfully generate a scFv to the highly carcinogenic toxin aflatoxin B1. Such an antibody has major potential applications in therapy and diagnostics. To further exploit its analytical capacity, the scFv was genetically fused to alkaline phosphatase, thereby generating a novel and highly sensitive self-indicating reagent. The performance of this reagent was further characterized, demonstrating its efficacy. The sensitivity of scFv-AP fusion was three-fold better than that of the scFv form. The ability of this human library to generate antibodies to a small hapten was clearly demonstrated and this is linked to its intrinsic diversity, which exceeds many existing conventional human libraries. Our results indicate that demography may influence the diversity of the repertoire of the library in terms of its capacity to generate antibodies to specific targets. Equally, the approach demonstrated should also be applicable for other haptens and larger antigens.

Rangnoi, Kuntalee; Jaruseranee, Nanthnit

2011-01-01

286

Kinetic of Alkaline Phosphatase in Liver, Kidney, Intestine and Muscle tissue of Red Tilapia Cultured Under Mid-Hill Altitudes of Meghalaya: North Eastern India  

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Full Text Available The aim of the present research was to provide information on the activities of Alkaline Phosphatase (ALP in various organs of Red Tilapia cultured under mid-hill condition of Meghalaya in captive condition, where within a year the water temperature ranges from 15-22°C. From the present study, it has been observed that, under mid-hill conditions of Meghalaya, the ALP activity was more in kidney (3983.6609±24.6838 mg L-1 and the decreasing order of activity was recorded in liver (3761.9639±18.6786 mg L-1, intestine (3420.1118±443.3330 mg L-1 and muscle tissue (1969.1100±22.5985 mg L-1. The higher distribution of ALP in kidney and other organs implies that the fish can adapt to cold temperature of mid-hill altitude and can be taken up as a candidate species for mass scale culture in mid-hill condition to meet the nutritional requirements of the people.

Sullip K. Majhi

2006-01-01

287

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

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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.

Benjamin Nnamdi Okolonkwo

2013-01-01

288

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

Science.gov (United States)

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. PMID:17437303

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

2007-11-01

289

Aedes aegypti alkaline phosphatase ALP1 is a functional receptor of Bacillus thuringiensis Cry4Ba and Cry11Aa toxins.  

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Bacillus thuringiensis subs. israelensis produces at least three Cry toxins (Cry4Aa, Cry4Ba, and Cry11Aa) that are active against Aedes aegypti larvae. Previous work characterized a GPI-anchored alkaline phosphatase (ALP1) as a Cry11Aa binding molecule from the gut of A. aegypti larvae. We show here that Cry4Ba binds ALP1, and that the binding and toxicity of Cry4Ba mutants located in loop 2 of domain II is correlated. Also, we analyzed the contribution of ALP1 toward the toxicity of Cry4Ba and Cry11Aa toxins by silencing the expression of this protein though RNAi. Efficient silencing of ALP1 was demonstrated by real-time quantitative PCR (qPCR) and Western blot. ALP1 silenced larvae showed tolerance to both Cry4Ba and Cry11Aa although the silenced larvae were more tolerant to Cry11Aa in comparison to Cry4Ba. Our results demonstrate that ALP1 is a functional receptor that plays an important role in the toxicity of the Cry4Ba and Cry11Aa proteins. PMID:22728570

Jiménez, Alan I; Reyes, Esmeralda Z; Cancino-Rodezno, Angeles; Bedoya-Pérez, Leidy P; Caballero-Flores, Gustavo G; Muriel-Millan, Luis F; Likitvivatanavong, Supaporn; Gill, Sarjeet S; Bravo, Alejandra; Soberón, Mario

2012-09-01

290

Identification of a selective inhibitor of murine intestinal alkaline phosphatase (ML260) by concurrent ultra-high throughput screening against human and mouse isozymes.  

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Alkaline phosphatase (AP) isozymes are present in a wide range of species from bacteria to man and are capable of dephosphorylation and transphosphorylation of a wide spectrum of substrates in vitro. In humans, four AP isozymes have been identified-one tissue-nonspecific (TNAP) and three tissue-specific-named according to the tissue of their predominant expression: intestinal (IAP), placental (PLAP) and germ cell (GCAP) APs. Modulation of activity of the different AP isozymes may have therapeutic implications in distinct diseases and cellular processes. For instance, changes in the level of IAP activity can affect gut mucosa tolerance to microbial invasion due to the ability of IAP to detoxify bacterial endotoxins, alter the absorption of fatty acids and affect ectopurinergic regulation of duodenal bicarbonate secretion. To identify isozyme selective modulators of the human and mouse IAPs, we developed a series of murine duodenal IAP (Akp3-encoded dIAP isozyme), human IAP (hIAP), PLAP, and TNAP assays. High throughput screening and subsequent SAR efforts generated a potent inhibitor of dIAP, ML260, with specificity for the Akp3-, compared to the Akp5- and Akp6-encoded mouse isozymes. PMID:24412070

Ardecky, Robert J; Bobkova, Ekaterina V; Kiffer-Moreira, Tina; Brown, Brock; Ganji, Santhi; Zou, Jiwen; Pass, Ian; Narisawa, Sonoko; Iano, Flávia Godoy; Rosenstein, Craig; Cheltsov, Anton; Rascon, Justin; Hedrick, Michael; Gasior, Carlton; Forster, Anita; Shi, Shenghua; Dahl, Russell; Vasile, Stefan; Su, Ying; Sergienko, Eduard; Chung, Thomas D Y; Kaunitz, Jonathan; Hoylaerts, Marc F; Pinkerton, Anthony B; Millán, José Luis

2014-02-01

291

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

Science.gov (United States)

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)2?2H2O (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.

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

2014-01-01

292

Expanding the spectrum of phenotypes associated with germline PIGA mutations: a child with developmental delay, accelerated linear growth, facial dysmorphisms, elevated alkaline phosphatase, and progressive CNS abnormalities.  

Science.gov (United States)

Phosphatidyl inositol glycan (PIG) enzyme subclasses are involved in distinct steps of glycosyl phosphatidyl inositol anchor protein biosynthesis. Glycolsyl phosphatidyl inositol-anchored proteins have heterogeneous functions; they can function as enzymes, adhesion molecules, complement regulators and co-receptors in signal transduction pathways. Germline mutations in genes encoding different members of the PIG family result in diverse conditions with (severe) developmental delay, (neonatal) seizures, hypotonia, CNS abnormalities, growth abnormalities, and congenital abnormalities as hallmark features. The variability of clinical features resembles the typical diversity of other glycosylation pathway deficiencies such as the congenital disorders of glycosylation. Here, we report the first germline missense mutation in the PIGA gene associated with accelerated linear growth, obesity, central hypotonia, severe refractory epilepsy, cardiac anomalies, mild facial dysmorphic features, mildly elevated alkaline phosphatase levels, and CNS anomalies consisting of progressive cerebral atrophy, insufficient myelinization, and cortical MRI signal abnormalities. X-exome sequencing in the proband identified a c.278C>T (p.Pro93Leu) mutation in the PIGA gene. The mother and maternal grandmother were unaffected carriers and the mother showed 100% skewing of the X-chromosome harboring the mutation. These results together with the clinical similarity of the patient reported here and the previously reported patients with a germline nonsense mutation in PIGA support the determination that this mutation caused the phenotype in this family. PMID:24259184

van der Crabben, Saskia N; Harakalova, Magdalena; Brilstra, Eva H; van Berkestijn, Frédérique M C; Hofstede, Floris C; van Vught, Adrianus J; Cuppen, Edwin; Kloosterman, Wigard; Ploos van Amstel, Hans Kristian; van Haaften, Gijs; van Haelst, Mieke M

2014-01-01

293

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  

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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 vari...

Hou, Guanhua; Cui, Qiang

2012-01-01

294

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

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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

Parker Bruce R

2009-07-01

295

Relationship between placental alkaline phosphatase activity and cord blood glucose, albumin and neonatal birth weight at term Relación entre la actividad de la fosfatasa alcalina placentaria, glucosa y albúmina en sangre del cordón umbilical y el peso en recién nacidos a término  

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It has been observed that placental alkaline phosphatase (PAP) activity progressively rises as pregnancy advances, possibly, because of its increasing synthesis by placental tissue. The present investigation therefore, examines the relationship between placental alkaline phosphatase activity and the biochemical indices of foetal nutrition (cord blood glucose, albumin) and growth (neonatal birth weight). Placental and umbilical cord blood samples were collected from one hundred and five delive...

Innocent Onyesom; Adefunke Olukemi Opajobi; Ugochukwu Enyinanya Uzuegbu; Denis Oriero; Joseph Mordi; Prosper Ejiro Awhin; Enaholo Timothy

2009-01-01

296

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  

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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 fol...

2012-01-01

297

Cadherin, alkaline phosphatase, and aminopeptidase N as receptors of Cry11Ba toxin from Bacillus thuringiensis subsp. jegathesan in Aedes aegypti.  

Science.gov (United States)

Cry11Ba is one of the most toxic proteins to mosquito larvae produced by Bacillus thuringiensis. It binds Aedes aegypti brush border membrane vesicles (BBMV) with high affinity, showing an apparent dissociation constant (K(d)) of 8.2 nM. We previously reported that an anticadherin antibody competes with Cry11Ba binding to BBMV, suggesting a possible role of cadherin as a toxin receptor. Here we provide evidence of specific cadherin repeat regions involved in this interaction. Using cadherin fragments as competitors, a C-terminal fragment which contains cadherin repeat 7 (CR7) to CR11 competed with Cry11Ba binding to BBMV. This binding was also efficiently competed by the CR9, CR10, and CR11 peptide fragments. Moreover, we show CR11 to be an important region of interaction with Cry11Ba toxin. An alkaline phosphatase (AaeALP1) and an aminopeptidase-N (AaeAPN1) also competed with Cry11Ba binding to Ae. aegypti BBMV. Finally, we found that Cry11Ba and Cry4Ba share binding sites. Synthetic peptides corresponding to loops ?8, ?2-?3 (loop 1), ?8-?9, and ?10-?11 (loop 3) of Cry4Ba compete with Cry11Ba binding to BBMV, suggesting Cry11Ba and Cry4Ba have common sites involved in binding Ae. aegypti BBMV. The data suggest that three different Ae. aegypti midgut proteins, i.e., cadherin, AaeALP1, and AaeAPN1, are involved in Cry11Ba binding to Ae. aegypti midgut brush border membranes. PMID:21037295

Likitvivatanavong, Supaporn; Chen, Jianwu; Bravo, Alejandra; Soberón, Mario; Gill, Sarjeet S

2011-01-01

298

Cadherin, Alkaline Phosphatase, and Aminopeptidase N as Receptors of Cry11Ba Toxin from Bacillus thuringiensis subsp. jegathesan in Aedes aegypti?  

Science.gov (United States)

Cry11Ba is one of the most toxic proteins to mosquito larvae produced by Bacillus thuringiensis. It binds Aedes aegypti brush border membrane vesicles (BBMV) with high affinity, showing an apparent dissociation constant (Kd) of 8.2 nM. We previously reported that an anticadherin antibody competes with Cry11Ba binding to BBMV, suggesting a possible role of cadherin as a toxin receptor. Here we provide evidence of specific cadherin repeat regions involved in this interaction. Using cadherin fragments as competitors, a C-terminal fragment which contains cadherin repeat 7 (CR7) to CR11 competed with Cry11Ba binding to BBMV. This binding was also efficiently competed by the CR9, CR10, and CR11 peptide fragments. Moreover, we show CR11 to be an important region of interaction with Cry11Ba toxin. An alkaline phosphatase (AaeALP1) and an aminopeptidase-N (AaeAPN1) also competed with Cry11Ba binding to Ae. aegypti BBMV. Finally, we found that Cry11Ba and Cry4Ba share binding sites. Synthetic peptides corresponding to loops ?8, ?2-?3 (loop 1), ?8-?9, and ?10-?11 (loop 3) of Cry4Ba compete with Cry11Ba binding to BBMV, suggesting Cry11Ba and Cry4Ba have common sites involved in binding Ae. aegypti BBMV. The data suggest that three different Ae. aegypti midgut proteins, i.e., cadherin, AaeALP1, and AaeAPN1, are involved in Cry11Ba binding to Ae. aegypti midgut brush border membranes.

Likitvivatanavong, Supaporn; Chen, Jianwu; Bravo, Alejandra; Soberon, Mario; Gill, Sarjeet S.

2011-01-01

299

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

Science.gov (United States)

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. PMID:16915846

Salter, Robert S; Fitchen, John

2006-01-01

300

Mosaic expression of pluripotency-related proteins oct-3/4 and alkaline phosphatase in human pancreatic carcinoma cell PANC-1  

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Full Text Available Most current research on cancer stem cells (CSCs associated with human tumorshas focused on the molecular and cellular analysis of hematopoietic lineagemarkers (e.g., CD44, CD138, and CD 133, which can also serve as important CSC markers in a variety of cancers. However, these markers are generallyexpressed at late stages in embryonic development. Oct-3/4, a member of thefamily of POU-domain transcriptional factors, and alkaline phosphatase (ALP areknown to be expressed in the inner cell mass of blastocysts, germ cells, andpluripotent embryonic stem cells. We thus consider Oct-3/4 and ALP to bepromising markers for CSC. Herein, we examined expression of Oct-3/4 and ALPusing 6 established human pancreatic carcinoma cell lines. RT-PCR analysisrevealed the presence of Oct-3/4 and ALP mRNA in those cells.Immunocytochemical and cytochemical staining revealed that both Oct-3/4 andALP proteins are present as mosaics in PANC-1 cell line, one of those 6 cell lines(23% and 19%, respectively. However, Oct-3/4-positive PANC-1 cells did notexhibit overt ATP-binding cassette transporter G2 (ABCG2 activity, as revealedby Hoechst 33342 dye exclusion assay. Transfection of PANC-1 cells with anOct-3/4 promoter-directed, enhanced green fluorescent protein (EGFP constructconfirmed the presence of Oct-3/4-positive cells. These findings indicate that inPANC-1 cells there are at least 2 subset populations, namely Oct-3/4-positive andALP-positive cells. However, it remains unknown whether expression of these 2markers overlaps. Enrichment of Oct-3/4- or ALP-positive cells by gene transferand subsequent drug selection will be helpful for further characterization of thesecells as possible CSCs.

Masahiro Sato

2013-01-01

 
 
 
 
301

Characterization of soluble and membrane-bound alkaline phosphatase in Nilaparvata lugens and their potential relation to development and insecticide resistance.  

Science.gov (United States)

Two forms (soluble and membrane-bound) of alkaline phosphatases (ALPs) were found in the brown planthopper, Nilaparvata lugens. In order to further study ALPs in N. lugens, two putative ALP genes (Nl-ALP1 and Nl-ALP2) were identified in this pest. Both Nl-ALP1 and Nl-ALP2 show approximately the same degree of sequence identity (40-50%) to other insect soluble and membrane-bound forms of ALP. Correlation of ALP activity and mRNA levels at different developmental stages, or following application of 20-hydroxyecdysone (20E) and insecticide fenvalerate, suggests that Nl-ALP1 and Nl-ALP2 might encode a soluble (sALP) and a membrane-bound ALP (mALP), respectively. Nl-ALP1-specific antibody Nl1-I detected only a specific band in soluble protein preparations and Nl-ALP2 specific antibody Nl2-I only detected a specific band in insoluble protein preparations, which provided conclusive linkages between Nl-ALP1 and a sALP and between Nl-ALP2 and a m ALP. Then, Nl-ALP1 was denoted as Nl-sALP for a sALP and Nl-ALP2 was denoted as Nl-mALP for a mALP. Only sALP activity and Nl-sALP mRNA level were induced by 20E and fenvalerate, which was confirmed by the density of specific band detected by Nl1-I in Sus strain with or without fenvalerate treatment. Additionally, the sALP activity, as well as Nl-sALP mRNA level, was significantly higher in a fenvalerate resistant population, compared with Sus strain. These results indicate that the sALP is more responsive to chemical stimulus, such as hormone and insecticide, and might play dual roles in development and insecticide tolerance. © 2011 Wiley Periodicals, Inc. PMID:21769927

Wang, Zengxia; Liu, Shuhua; Yang, Baojun; Liu, Zewen

2011-07-18

302

Determination of trace alkaline phosphatase by affinity adsorption solid substrate room temperature phosphorimetry based on wheat germ agglutinin labeled with 8-quinolineboronic acid phosphorescent molecular switch and prediction of diseases  

Science.gov (United States)

The 8-quinolineboronic acid phosphorescent molecular switch (abbreviated as PMS-8-QBA. Thereinto, 8-QBA is 8-quinolineboronic acid, and PMS is phosphorescent molecular switch) was found for the first time. PMS-8-QBA, which was in the "off" state, could only emit weak room temperature phosphorescence (RTP) on the acetyl cellulose membrane (ACM). However, PMS-8-QBA turned "on" automatically for its changed structure, causing that the RTP of 8-QBA in the system increased, after PMS-8-QBA-WGA (WGA is wheat germ agglutinin) was formed by reaction between -OH of PMS-8-QBA and -COOH of WGA. More interesting is that the -NH 2 of PMS-8-QBA-WGA could react with the -COOH of alkaline phosphatase (AP) to form the affinity adsorption (AA) product WGA-AP-WGA-8-QBA-PMS (containing -NH-CO- bond), which caused RTP of the system to greatly increase. Thus, affinity adsorption solid substrate room temperature phosphorimetry using PMS-8-QBA as labelling reagent (PMS-8-QBA-AA-SSRTP) for the determination of trace AP was established. The method had many advantages, such as high sensitivity (the detection limit (LD) was 2.5 zg spot -1. For sample volume of 0.40 ?l spot -1, corresponding concentration was 6.2 × 10 -18 g ml -1), good selectivity (the allowed concentration of coexisting material was higher, when the relative error was ±5%), high accuracy (applied to detection of AP content in serum samples, the result was coincided with those obtained by enzyme-linked immunoassay), which was suitable for the detection of trace AP content in serum samples and the forecast of human diseases. Meanwhile, the mechanism of PMS-8-QBA-AASSRTP was discussed. The new field of analytical application and clinic diagnosis technique of molecule switch are exploited, based on the phosphorescence characteristic of PMS-8-QBA, the AA reaction between WGA and AP, as well as the relation between AP content and human diseases. The research results promote the development and interpenetrate among molecule switch technique, lectin science and SSRTP.

Liu, Jia-Ming; Gao, Hui; Li, Fei-Ming; Shi, Xiu-Mei; Lin, Chang-Qing; Lin, Li-Ping; Wang, Xin-Xing; Li, Zhi-Ming

2010-09-01

303

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  

International Nuclear Information System (INIS)

Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: ? The scFv-AP fusion protein against ractopamine (RAC) was produced. ? A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. ? The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. ? Recovery tests from pork samples were studied. ? 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 (VH and VL) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling VH and VL 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?1, respectively, and the linear response range extended from 0.05 to 1.45 ng mL?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 the data of dc-CLEIA and HPLC–MS (R2 > 0.99), indicating that the assay was an efficient analytical method for monitoring food safety.

2012-07-29

304

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  

Energy Technology Data Exchange (ETDEWEB)

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 the data of dc-CLEIA and HPLC-MS (R{sup 2} > 0.99), indicating that the assay was an efficient analytical method for monitoring food safety.

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: gzwhongd@63.com [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China)

2012-07-29

305

Polymer-anchored peroxo compounds of vanadium(V) and molybdenum(VI): synthesis, stability, and their activities with alkaline phosphatase and catalase.  

Science.gov (United States)

We generated a series of new polymer-bound peroxo complexes of vanadium(V) and molybdenum(VI) of the type [VO(O(2))(2)(sulfonate)]-PSS [PSS = poly(sodium 4-styrene sulfonate)] (PV(3)), [V(2)O(2)(O(2))(4)(carboxylate)VO(O(2))(2)(sulfonate)]-PSSM [PSSM = poly(sodium styrene sulfonate-co-maleate)] (PV(4)), [Mo(2)O(2)(O(2))(4)(carboxylate)]-PA [PA = poly(sodium acrylate)] (PMo(1)), [MoO(O(2))(2)(carboxylate)]-PMA [PMA = poly(sodium methacrylate)] (PMo(2)), and [MoO(O(2))(2)(amide)]-PAm [PAm = poly(acrylamide)] (PMo(3)) by reacting V(2)O(5) (for PV(3) and PV(4)) or H(2)MoO(4) (for PMo(1), PMo(2), and PMo(3)) with H(2)O(2) and the respective water-soluble macromolecular ligand at pH 5-6. The compounds were characterized by elemental analysis (CHN and energy-dispersive X-ray spectroscopy), spectral studies (UV-vis, IR, (13)C NMR, (51)V NMR, and (95) Mo NMR), thermal (TGA) as well as scanning electron micrographs (SEM), and EDX analysis. It has been demonstrated that compounds retain their structural integrity in solutions of a wide range of pH values and are approximately 100 times weaker as substrate to the enzyme catalase relative to H(2)O(2), its natural substrate. The effect of the title compounds, along with previously reported compounds [V(2)O(2)(O(2))(4)(carboxylate)]-PA (PV(1)) and [VO(O(2))(2)(carboxylate)]-PMA (PV(2)) on rabbit intestine alkaline phosphatase (ALP) has been investigated and compared with the effect induced by the free diperoxometallates viz. Na[VO(O(2))(2)(H(2)O)] (DPV), [MoO(O(2))(2)(glycine)(H(2)O)] (DMo(1)), and [MoO(O(2))(2)(asparagine)(H(2)O)] (DMo(2)). It has been observed that although all the compounds tested are potent inhibitors of the enzyme, the polymer-bound and neat complexes act via distinct mechanisms. Each of the macromolecular compounds is a classical noncompetitive inhibitor of ALP. In contrast, the action of neat pV and heteroligand pMo compounds on the enzyme function is consistent with a mixed type of inhibition. PMID:21786762

Boruah, Jeena Jyoti; Kalita, Diganta; Das, Siva Prasad; Paul, Saurav; Islam, Nashreen S

2011-09-01

306

Adsorption kinetics and dilatational rheological studies for the soluble and anchored forms of alkaline phosphatase at the air/water interface  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Este trabalho apresenta aspectos de equilíbrio e dinâmicos da adsorção na interface ar/líquido de duas formas de fosfatase alcalina de placa óssea de ratos: DSAP, solubilizada com tensoativo não-iônico (C12E9), contendo uma âncora de glicosilfosfatidilinositol (GPI), e PLSAP, com a porção hidrofóbic [...] a da âncora clivada por fosfolipase-C. A tensão superficial dinâmica, gamadyn, e o módulo de elasticidade superficial dilatacional, épsilon, foram determinados para soluções de PLSAP, DSAP e C12E9 pelo método de oscilação harmônica e análise do formato da gota eixo-simétrica. Cinéticas de adsorção revelaram que DSAP adsorve trinta vezes mais rapidamente que PLSAP, apresentando um mínimo, e, para PLSAP, a tensão superficial cai continuamente. Para o sistema DSAP/C12E9, épsilon atinge um máximo na concentração crítica de agregação (CAC), mas para PLSAP, épsilon diminui continuamente com a concentração. Soluções de C12E9 apresentam épsilonmais elevados, decrescentes com a concentração. Um modelo, baseado na influência da âncora GPI, é proposto para explicar os resultados obtidos. Abstract in english This work presents equilibrium and dynamic aspects for the adsorption at the air/liquid interface of two rat osseous plate alkaline phosphatase forms: DSAP, solubilized by a surfactant, C12E9, and containing a glycosylphosphatidylinositol (GPI) anchor; and PLSAP, resulting from phospholipase-C cleav [...] age of the hydrophobic portion of the GPI anchor. Dynamic surface tension, gammadyn, and surface elasticity modulus, epsilon, were determined for PLSAP, DSAP and pure C12E9 solutions using harmonic oscillation and axisymmetric drop shape analysis Adsorption kinetics studies revealed that DSAP adsorbs thirty times faster than PLSAP, presenting a minimum in the curve. For DSAP/ C12E9 mixed system, e increases with concentration and a maximum appears at the critical aggregation concentration (CAC). For PLSAP, a continuous decreasing with concentration for gammadyn and epsilon was observed. For pure C12E9 solution, the elasticity modulus increases with concentration and epsilon values are higher when compared to the mixed system. A model based on the influence of the GPI anchor is proposed.

Caseli, Luciano; Masui, Douglas C.; Furriel, Rosa Prazeres M.; Leone, Francisco Assis; Zaniquelli, Maria Elisabete D..

307

Site-specific mutations in the COOH-terminus of placental alkaline phosphatase: a single amino acid change converts a phosphatidylinositol- glycan-anchored protein to a secreted protein  

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Placental alkaline phosphatase (PLAP) is anchored in the plasma membrane by a phosphatidylinositol-glycan moiety (PI-glycan). PI-glycan is added posttranslationally to the nascent peptide chain after the removal of 29 amino acids from the COOH-terminus. The contribution of selected COOH-terminal amino acids to the signal for PI-glycan addition was tested by creating a fusion protein with the COOH-terminus of PLAP and a secreted protein and by mutagenesis of specific PLAP COOH- terminal amino ...

1992-01-01

308

Correlation of Serum Magnesium with Serum Parathormone in Regular Hemodialysis Patients  

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Full Text Available To consider the relationship of serum magnesium with the activity of the parathyroid gland in maintenance hemodialysis patients we designed a study to investigate the role of serum magnesium in regulating the parathyroid secretion. The study was conducted on patients undergoing maintenance hemodialysis treatment. Predialysis serum calcium, phosphorus, magnesium, alkaline phosphatase, intact serum PTH (iPTH, serum 25-hydroxy vitamin D (25-OH Vit D and plasma HCO3– were measured. The Urea Reduction Rate, duration and dosage of hemodialysis treatment were calculated also. In this study no significant correlation of serum magnesium with duration of hemodialysis treatment, alkaline phosphatase, plasma HCO3–, serum calcium and phosphorus patients were seen. In all patients a near significant inverse correlation of serum magnesium with iPTH (r = -0.30 , p = 0.079 was found, also a significant positive correlation of serum magnesium with serum 25-hydroxy vitamin D levels ( r = 0.40, p = 0.009 was seen. Earlier research concluded that some factors other than serum magnesium may be more important in the regulation of parahormone secretion in hemodialysis patients. A positive and strong association between serum magnesium with 25-hydroxy vitamin D level, needs to more attention to this aspect of hemodialysis patients.

Hamid Nasri

2006-01-01

309

Relação tireóide-gônadas e níveis plasmáticos de fósforo, cálcio e fosfatase alcalina em ratas Relationship between thyroid, gonads and plasmatic levels of phosphorus, calcium and alkaline phosphatase in rats  

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Full Text Available A relação tireóide-gônadas-metabolismo ósseo foi estudada em ratas Wistar adultas, castradas ou intactas e mantidas em estado hipertireóideo ou eutireóideo por períodos de 30, 60 e 90 dias. Foram utilizadas como características do metabolismo ósseo o cálcio, o fósforo e a atividade da fosfatase alcalina plasmáticos, correlacionando-os com os valores de estrógeno, de progesterona e de T4 livre. Verificou-se que o hipogonadismo e o hipertireoidismo alteram as características plasmáticas do metabolismo ósseo. O hipertireoidismo induz hiperfosfatemia e hipocalcemia, o hipogonadismo tem pouca influência sobre o fósforo, mas potencializa a hiperfosfatemia e a hipocalcemia desencadeadas pelo hipertireoidismo. Com relação à fosfatase alcalina, conclui-se que o hipertireoidismo reduz o efeito do hipogonadismo sobre a atividade dessa enzima.The interrelation between thyroid, gonads and osseous metabolism was studied in either intact or castrated adult female rats kept under hyperthyroidism or euthyroidism for 30, 60, or 90 days. Plasmatic levels of phosphorus, calcium, and alkaline phosphatase were measured to assess the osseous metabolism. These characteristics were correlated to the levels of estrogen, progesterone, and free T4. Either hypogonadism or hyperthyroidism interfered with the plasmatic characteristics of osseous metabolism. Hyperphosphatemia and hypocalcemia were induced by hyperthyroidism, whereas the hypogonadism had little effect on the levels of phosphorus, but it had a potencialization effect on the hyperphosphatemia and hypocalcemia induced by hyperthyroidism. The effect of hypogonadism on the alkaline phosphatase activity was reduced by the hyperthyroidism.

R. Serakides

2000-12-01

310

Serum tartrate-resistant acid phosphatase 5b activity as a prognostic marker of survival in breast cancer with bone metastasis  

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Full Text Available Abstract Background Serum tartrate-resistant acid phosphatase 5b (TRACP 5b activity is a marker of osteoclast number and is elevated in breast cancer (BC patients with extensive bone metastasis, which might in turn reflect the tumour burden. We tested the hypothesis that baseline serum TRACP 5b activity and its interval change are potential prognostic markers of survival in BC patients with bone metastasis. Methods We analyzed the data from previous prospective studies. A total of 100 patients with newly diagnosed bone metastasis were included. Cox proportional regression model was used to evaluate the correlation between the overall survival time (OS and baseline serum TRACP 5b activity and its interval changes. The least significant change (LSC of TRACP 5b was calculated from data obtained from 15 patients with early BC. Results Estrogen receptor status (Hazard Ratio (HR = 0.397; p = 0.003 and visceral metastasis (HR = 0.492; p = 0.0045 were significantly correlated with OS. The OS was significantly shorter in those patients with higher baseline TRACP 5b activity based on a cut-off value to delineate the highest tertile (HR = 3.524; p p = 0.0015. Conclusions We found that TRACP 5b activity and its interval change after treatment bore a prognostic role in BC patients with bone metastasis and a high baseline serum TRACP 5b activity. Further prospective phase II study is necessary to confirm these results.

Liu Hsin-Yi

2010-04-01

311

Effects of Aqueous Stem-Bark Extract of Momordica balsamina Linn on Some Serum Enzymes in Normal and Ethanol Fed Rats  

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Aqueous stem-bark extract of Momordica balsamina Linn was administered using stomach tubes to normal and alcohol fed rats to study the effect of the extract on organs and tissues by estimating the level of some serum enzymes. The extract was administered for two weeks at a dose of 0.56 mg/100 g body weight. The parameters studied include some serum enzymes (Prostatic and total acid phosphatase, alkaline phosphatase, alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT))...

Geidam, M. A.; Dauda, E.; Hamza, H. G.

2007-01-01

312

Serum and pancreatic juice carcinoembryonic antigen in pancreatic and biliary disease.  

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Serum and pancreatic juice carcinoembryonic antigen (CEA) concentrations were studied in a group of 144 patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) with a variety of benign and malignant pancreatic and biliary diseases. Serum CEA was found to be a poor diagnostic and discriminating marker for pancreatic disorders and was raised in obstructive jaundice from various causes correlating with serum alkaline phosphatase. A pancreatic juice CEA concentration of greater ...

Carr-locke, D. L.

1980-01-01

313

Relação tireóide-gônadas e níveis plasmáticos de fósforo, cálcio e fosfatase alcalina em ratas / Relationship between thyroid, gonads and plasmatic levels of phosphorus, calcium and alkaline phosphatase in rats  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese A relação tireóide-gônadas-metabolismo ósseo foi estudada em ratas Wistar adultas, castradas ou intactas e mantidas em estado hipertireóideo ou eutireóideo por períodos de 30, 60 e 90 dias. Foram utilizadas como características do metabolismo ósseo o cálcio, o fósforo e a atividade da fosfatase alca [...] lina plasmáticos, correlacionando-os com os valores de estrógeno, de progesterona e de T4 livre. Verificou-se que o hipogonadismo e o hipertireoidismo alteram as características plasmáticas do metabolismo ósseo. O hipertireoidismo induz hiperfosfatemia e hipocalcemia, o hipogonadismo tem pouca influência sobre o fósforo, mas potencializa a hiperfosfatemia e a hipocalcemia desencadeadas pelo hipertireoidismo. Com relação à fosfatase alcalina, conclui-se que o hipertireoidismo reduz o efeito do hipogonadismo sobre a atividade dessa enzima. Abstract in english The interrelation between thyroid, gonads and osseous metabolism was studied in either intact or castrated adult female rats kept under hyperthyroidism or euthyroidism for 30, 60, or 90 days. Plasmatic levels of phosphorus, calcium, and alkaline phosphatase were measured to assess the osseous metabo [...] lism. These characteristics were correlated to the levels of estrogen, progesterone, and free T4. Either hypogonadism or hyperthyroidism interfered with the plasmatic characteristics of osseous metabolism. Hyperphosphatemia and hypocalcemia were induced by hyperthyroidism, whereas the hypogonadism had little effect on the levels of phosphorus, but it had a potencialization effect on the hyperphosphatemia and hypocalcemia induced by hyperthyroidism. The effect of hypogonadism on the alkaline phosphatase activity was reduced by the hyperthyroidism.

R., Serakides; V.A., Nunes; E.F., Nascimento; C.M., Silva; A.F.C., Ribeiro.

314

Effects of Aqueous Stem-Bark Extract of Momordica balsamina Linn on Some Serum Enzymes in Normal and Ethanol Fed Rats  

Directory of Open Access Journals (Sweden)

Full Text Available Aqueous stem-bark extract of Momordica balsamina Linn was administered using stomach tubes to normal and alcohol fed rats to study the effect of the extract on organs and tissues by estimating the level of some serum enzymes. The extract was administered for two weeks at a dose of 0.56 mg/100 g body weight. The parameters studied include some serum enzymes (Prostatic and total acid phosphatase, alkaline phosphatase, alanine aminotransferase (ALAT and aspartate aminotransferase (ASAT, serum glucose, albumin and total protein. Results obtained shows that the stem-bark extract has hypoglycaemic effect in rats. The extract alone was observed to have significant effect on alkaline phosphatase. The level of albumin was insignificantly increased as well as those of ALAT and ASAT. Prostatic and total acid phosphatases were observed to be significantly increased in ethanol fed rats alone also.

M.A. Geidam

2007-01-01

315

Diagnosis of testicular carcinoma in situ, (intratubular- and micro-invasive) seminoma and embryonal carcinoma using direct enzymatic alkaline phosphatase reactivity on frozen histological sections.  

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Abstract Aims: Testis-sparing surgery might benefit quality of life, but can only be applied with histological examination for seminoma and embryonal carcinoma, and carcinoma in situ (CIS). Diagnosis is based on paraffin-embedded tissue, therefore a delay in further surgery is mostly unavoidable. Methods and Results: A total of 4,093 snap frozen samples and paraffin tissue of 1,500 patients were included. Besides standard H & e staining, the direct enzymatic alkaline phosphatas...

2011-01-01

316

Variation of pH in lysed platelet concentrates influence proliferation and alkaline phosphatase activity in human osteoblast-like cells.  

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Activated platelets release a multifaceted blend of growth factors that has stimulatory effects on mesenchymal cells, both in vitro and in vivo, which imply beneficial effects on wound repair and tissue regeneration. Previous studies on fibroblast cultures have revealed that more potent growth factors, with respect to cell proliferation, are released in acidic preparations of lysed platelet concentrates in comparison with neutral and alkaline preparations. The current study was intended to in...

Wahlstro?m, Ola; Linder, Cecilia; Kale?n, Anders; Magnusson, Per

2007-01-01

317

The dermatophyte Trichophyton rubrum secretes an EDTA-sensitive alkaline phosphatase on high-phosphate medium O dermatófito Trichophyton rubrum secreta uma fosfatase alcalina EDTA-sensível em meio contendo alta concentração de fosfato  

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Full Text Available In this communication, we show that the growth of isolate H6 of the dermatophyte Trichophyton rubrum on non-buffered medium and under saturating phosphate conditions is dependent on the initial growth pH, with an apparent optimum at pH 4.0. In addition, irrespective of the initial growth pH, the pH of the medium alteredduring cultivation reaching values that ranged from 8.3 to 8.9. Furthermore, this isolate synthesized and secreted almost the same levels of an alkaline phosphatase with an apparent optimum pH ranging from 9.0 to 10.0 when grown on both low- and high-phosphate medium. Also, this alkaline phosphatase is activated by Mg2+ and is EDTA-sensitive. On the other hand, the very low levels of the enzyme retained by the mycelium grown on buffered medium at pH 5.0-5.2 suggest that this enzyme is encoded by an alkaline gene, i.e., a gene responsive to ambient pH signaling.Nesta comunicação nós mostramos que o crescimento do isolado H6 do dermatófito T. rubrum em meio não tamponado e sob condição saturante de fosfato, é dependente do pH inicial de cultivo, com um ótimo aparente em pH 4,0. Além disto, independente do pH inicial, o pH do meio se altera durante o cultivo alcançando valores que variam de 8,3 a 8,9. Verificou-se também que este isolado sintetiza e secreta quase os mesmos níveis de fosfatase alcalina, com um ótimo de atividade aparente entre os valores de pH 9,0 e 10,0, independentemente da concentração de fosfato no meio. Também mostramos que essa fosfatase alcalina é inibida por EDTA e ativada por Mg2+. Por outro lado, o nível dessa enzima retida no micélio cultivado em meio tamponado em pH 5,0-5,2 é baixo, sugerindo que ela seja codificada por um gene alcalino, isto é, um gene responsivo à sinalização pelo pH ambiente.

Monica S. Ferreira-Nozawa

2003-06-01

318

SERUM CHEMISTRIES OF COTURNIX JAPONICA GIVEN DIETARY MANGANESE OXIDE (MN3O4)  

Science.gov (United States)

Plasma creatinine and inorganic phosphorus were increased in manganese oxide (Mn3O4)-treated adult male Coturnix quail, but BUN, BUN/creatinine ratio, uric acid, and total calcium were decreased. 2. Serum enzymes (alkaline phosphatase glutamic oxaloacetic transaminase, glutamic p...

319

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

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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...

Prl, Silva; Oc, Freitas Neto; Ac, Laurentiz; Om, Junqueira; Jj, Fagliari

2007-01-01

320

Effect of Refined Petroleum Product (Kerosene Flame and Fumes on Serum Enzyme Characteristics of Broiler Chickens  

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Full Text Available Enzyme serology of broiler chickens exposed to refined petroleum product (kerosene flame and fumes at varying distances over a period of 16 hrs daily for 56 days in a poultry house was evaluated. The burning of kerosene was simulated in a designed burner. The measured distances were 4m, 8m and 12m from the flame point. The control birds were located in a separate poultry building without the flame treatment. Proprietary broiler starter and finisher diets were fed ad libitum. Blood samples were taken at the 4th and 8th weeks for enzymological assay from each treatment. The enzymes assayed were Serum Glutamic Oxaloacetic Transaminase (SGOT, Serum Glutamic Pyruvic Transaminase (SGPT and Alkaline Phosphatase (ALP. No significant (P > 0.05 differences were observed in Serum Glutamic Pyruvic Transaminase (SGPT and Alkaline Phosphatase (ALP. Results also showed that there were significant differences (P < 0.05 in Serum Glutamic Oxaloacetic Transaminase (SGOT.

A.O. Amakiri

2008-01-01

 
 
 
 
321

Use of progesterone 11-glucuronide-alkaline phosphatase conjugate in a sensitive microtitre-plate enzymeimmunoassay of progesterone in milk and its application to pregnancy testing in dairy cattle.  

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A simple direct-addition microtitre plate enzymeimmunoassay (EIA) for progesterone in whole milk is described. The assay used antiserum raised against 11 alpha-hydroxyprogesterone 11-hemisuccinate (progesterone 11-hemisuccinate) and a heterologous label prepared by conjugation of 11 alpha-hydroxyprogesterone 11-glucuronide (progesterone 11-glucuronide) with alkaline phosphatase using an active ester procedure. The sensitivity, analytical recovery, linearity of response and precision of the assay compared favourably with radioimmunoassay (RIA). Results from EIA of milk samples were compared with determinations made after isolation of progesterone by HPLC (r = 0.910). Milk samples (200) were assayed by RIA at both the Milk Marketing Board and the Cattle Breeding Centre and the results were correlated with EIA performed at the Cattle Breeding Centre (r = 0.890 and r = 0.833 respectively). Calving data were obtained from a further 110 cows for which the milk progesterone EIA had provided a pregnancy test 24 days after AI; 46 cows were correctly identified as non-pregnant and 58 as pregnant and there were 4 false positive and 2 inconclusive results. PMID:3511256

Sauer, M J; Foulkes, J A; Worsfold, A; Morris, B A

1986-01-01

322

Cercospora beticola toxins. Part XVII. The role of the beticolin/Mg2+ complexes in their biological activity. Study of plasma membrane H(+)-ATPase, vacuolar H(+)-PPase, alkaline and acid phosphatases.  

Science.gov (United States)

Beticolin-1 and beticolin-2, yellow toxins produced by the phytopathogenic fungus Cercospora beticola, inhibit the plasma membrane H(+)-ATPase. Firstly, since beticolins are able to form complexes with Mg2+, the role of the beticolin/Mg2+ complexes in the inhibition of the plasma membrane proton pump has been investigated. Calculations indicate that beticolins could exist under several forms, in the H(+)-ATPase assay mixture, both free or complexed with Mg2+. However, the percentage inhibition of the H(+)-ATPase activity is correlated to the concentration of one single form of beticolin, the dimeric neutral complex Mg2H2B2, which appears to be the active form involved in the H(+)-ATPase inhibition. Secondly, since previous data suggested that beticolins could also be active against other Mg2(+)-dependent enzymes, we tested beticolin-1 on the vacuolar H(+)-PPase, which requires Mg2+ as co-substrate, and on the alkaline and acid phosphatases, which do not use Mg2+ as co-substrate. Only vacuolar H(+)-PPase is sensitive to beticolin-1, which suggests that beticolins are specific to enzymes that use a complex of Mg2+ as the substrate. The same Mg2H2B2 complex which is responsible of the plasma membrane H(+)-ATPase inhibition appears to be also involved in the inhibition of the vacuolar H(+)-PPase. PMID:8948473

Gomès, E; Gordon-Weeks, R; Simon-Plas, F; Pugin, A; Milat, M L; Leigh, R A; Blein, J P

1996-11-13

323

Yu Ping Feng San, an Ancient Chinese Herbal Decoction, Regulates the Expression of Inducible Nitric Oxide Synthase and Cyclooxygenase-2 and the Activity of Intestinal Alkaline Phosphatase in Cultures  

Science.gov (United States)

Yu Ping Feng San (YPFS), a Chinese herbal decoction comprising Astragali Radix (AR; Huangqi), Atractylodis Macrocephalae Rhizoma (AMR; Baizhu), and Saposhnikoviae Radix (SR; Fangfeng), has been used clinically to treat inflammatory bowel diseases (IBD). Previously, we demonstrated a dual role of YPFS in regulating cytokine release in cultured macrophages. In this study, we elucidated the anti-inflammatory effect of YPFS that is mediated through modulating the expression of three key enzymes involved in IBD: inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and intestinal alkaline phosphatase (IALP). In a lipopolysaccharide (LPS)-induced chronic-inflammation model of cultured murine macrophages, YPFS treatment suppressed the activation of iNOS and COX-2 expression in a dose-dependent manner. Conversely, application of YPFS in cultured small intestinal enterocytes markedly induced the expression of IALP in a time-dependent manner, which might strengthen the intestinal detoxification system. A duality of YPFS in modulating the expression of iNOS and COX-2 was determined here. The expression of iNOS and COX-2 in macrophages was induced by YPFS, and this activation was partially blocked by the NF-?B-specific inhibitor BAY 11-7082, indicating a role of NF-?B signaling. These YPFS-induced changes in gene regulation strongly suggest that the anti-inflammatory effects of YPFS are mediated through the regulation of inflammatory enzymes.

Du, Crystal Y. Q.; Choi, Roy C. Y.; Dong, Tina T. X.; Lau, David T. W.; Tsim, Karl W. K.

2014-01-01

324

Serum dickkopf-1 level in postmenopausal females: correlation with bone mineral density and serum biochemical markers.  

Science.gov (United States)

Objective. To assess serum level of Dickkopf-1 in postmenopausal females and its correlation with bone mineral density and serum biochemical markers. Methods. Bone densitometry, serum Dickkopf-1, calcium, phosphorus, and alkaline phosphatase were done in sixty postmenopausal females. Patients were divided according to T score into osteoporosis (group I), osteopenia (group II), and normal bone mineral density that served as controls. Results. There was highly significant increase in serum Dickkopf-1 levels in postmenopausal females with abnormal T score versus controls (P 0.05). Conclusion. Postmenopausal females may suffer from osteoporosis as evidenced by bone densitometry. Postmenopausal women with significantly increased serum Dickkopf-1 had more significant osteoporosis. Prolonged duration of menopause and increased serum Dickkopf-1 are important risk factors for the development and severity of osteoporosis. PMID:23878759

Ahmed, Sahar Fathi; Fouda, Neveen; Abbas, Amal Ahmed

2013-01-01

325

The Predictivity of Serum Biochemical Markers in Acute Biliary Pancreatitis  

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Background and Aim. There are no accurate methods of differentiating acute biliary pancreatitis. Obstructions of biliary ducts, idiopathic pancreatitis may be related with biliary origin which needs identification for acute treatment. We searched for the predictivity of biochemical markers in early acute biliary pancreatitis. Patients and Methods. Serum levels of AST (Aspartate Transaminase),ALT (Alanine Transaminase), ALP (Alkaline Phosphatase), GGT (Gamma Glutamyl Transferase), total bilir...

Gu?ngo?r, Bu?lent; C?ag?layan, Kas?m; Polat, Cafer; S?eren, Deniz; Erzurumlu, Kenan; Malazgirt, Zafer

2011-01-01

326

Serum biochemical values of farmed ostrich (Struthio camelus) in Botswana.  

Science.gov (United States)

Reference biochemical values for serum analytes of 126 clinically normal farmed ostriches on one farm in Botswana were established. These included sodium, potassium, chloride, total protein, albumin, urea, creatinine, uric acid, cholesterol, total bilirubin, conjugated bilirubin, glucose, triglyceride, calcium, phosphorus, manganese, copper, zinc, alkaline phosphatase, gamma glutamyl transferase and creatinine kinase. The values obtained in this study can be used as reference values. PMID:9809323

Mushi, E Z; Binta, M G; Chabo, R G; Isa, J F; Modisa, L

1998-09-01

327

Introducción del método inmunocitoquímico de la fosfatasa alcalina-antifosfatasa alcalina para la clasificación inmunológica de los Síndromes Linfo y Mieloproliferativos Agudos / Introduction of the alkaline phosphatase-alkaline antiphosphatase immunocytochemical method for the immunological classification of the acute lympho-and myeloproliferative syndromes  

Scientific Electronic Library Online (English)

Full Text Available SciELO Cuba | Language: Spanish Abstract in spanish Se realizó el inmunofenotipaje celular en 30 pacientes con el diagnóstico de síndromes linfo y mieloproliferativos agudos por el método inmunoenzimático fosfatasa alcalina-antifosfatasa alcalina (APAAp) introducido en nuestro laboratorio. Los marcadores estudiados fueron: CD3, CD5, CD7, CD10, CD13, [...] CD15, CD22, CD33, CD34 y CD41 mediante los anticuerpos monoclonales correspondientes, según cada caso. De las leucemias agudas, 16 resultaron ser leucemias linfoides (LLA) (53,3 %) y 12 mieloides (LMA) (40 %). Entre las LLA, el 50 % fue del fenotipo B y del resto, 1 caso del tipo T (LLA-T) (3,33 %). Un paciente se diagnosticó como leucemia aguda híbrida (LAH) (3,33 %) y el otro se clasificó como leucemia aguda indiferenciada (LAI) (3,33 %). Se concluye que el APAAP es un método más rápido y tan eficaz como otros métodos enzimáticos para la clasificación inmunológica de los síndromes linfo y mieloproliferativos Abstract in english The cellular immunophenotyping was carried out in 30 patients with the diagnosis of acute lympho- and myeloproliferative syndromes by the alkaline phosphatase-alkaline antiphosphatase immunoenzimatic method (APAA) introduced in our laboratory. The CD3, CD5, CD7, CD10; CDl3, CDl5, CD22, CD33 and CD41 [...] markers were studied by using the corresponding monoclonal antibodies, according to each case. Of the acute leukemias, 16 were lymphoid leukemias (ALL) (53.3 %) and 12 were myeloid leukemias (AML) (40 %). Among the ALL, 50 % were phenotype B and of the rest, 1 case was type T (ALL-T) (3.33 %). A patient was diagnosed acute hybrid leukemia (AHL) (3.33 %) and the other was classified as acute undifferentiated leukemia (AUL) (3.33 %). It is concluded that the APAA is faster and as efficient as other enzimatic methods for the immunologic classification of the lympho- and myeloproliferative syndromes

Berta B, Socarrás Ferrer; Vianed, Marsán Suárez; Miriam, Sánchez Segura; Consuelo, Macías Abraham.

328

Elevated levels of serum type I collagen C-telopeptide in patients with rapidly destructive osteoarthritis of the hip  

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We compared type I collagen degradation using serum cross-linking C-terminal telopeptide (ICTP) in 18 patients with rapidly destructive osteoarthrosis and in 20 patients with slowly progressive osteoarthrosis of the hip. The diagnosis was established by clinical examination and radiographic evaluation. Total hip arthroplasty was performed in all patients. Serum levels of ICTP, bone-specific alkaline phosphatase, osteocalcin and N-terminal propeptide were studied. Patients with rapidly destruc...

Berger, Christian E.; Kro?ner, Andreas; Stiegler, Helmar; Leitha, Thomas; Engel, Alfred

2005-01-01

329

Study of serum lipase, alpha-amylase and pancreatic amylose in gall-stone diseases.  

Science.gov (United States)

Silent gall-stone causes significant morbidity and mortality and its incidence in India as well as in whole world is on the rise. It has positive correlation with development of carcinoma gall bladder. So far no predictive study has been done to show its correlation with biochemical markers. The present study has been aimed to establish whether simple enzymatic markers can predict association with cholelithiasis. Study group has been selected from the patients attending general surgery OPD of a tertiary healthcare centre with complaints of vague abdominal pain, flatulence and dyspepsia. A total of 61 cases (male = 18, female = 43) were studied and data matched with age and sex matched control. The biochemical markers studied are serum alkaline phosphatase, serum lipase, serum alpha-amylase and serum pancreatic amylase. Patients with obstructive cholelithiasis, duct stones, pancreatic insufficiency and malignancy are excluded from the study. The results were analysed by Student's t-test. Alkaline phosphatase in all the above mentioned cases was not significantly different from the control group (40 female, 21 male healthy individuals). A significant association was found out with serum alpha-amylase (p gall-stone diseases which are in many cases silent preventing further complications and chances of Malignancy especially where alkaline phosphatase isinconclusive. PMID:22480101

Bera, Swati; Bhattacharyya, Swati; Ghose, Bikash C; Bera, Tapas; Mukhopadhyay, Surajit K; Saha, Mita

2011-09-01

330

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

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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 Resear...

Abedini F; Hosseinkhani H; Ismail M.; Aj, Domb; Ar, Omar; Pp, Chong; Pd, Hong; Ds, Yu; Iy, Farber

2012-01-01

331

Crystallographic studies on shrimp alkaline phosphatase  

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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 have been found to be more efficient at low temperatures and less stable than enzymes from organisms living at moderate temperatures (such as mammals). In this study, the structure of a cold-acti...

Backer, M. M. E.

2003-01-01

332

Radioimmunoassay of serum glycocholic acid, standard laboratory tests of liver function and liver biopsy findings: comparative study of children with liver disease.  

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Serum glycocholic acid (SGC) was measured by radioimmunoassay in 277 samples from 122 children with hepatobiliary disorders and from 23 healthy age-matched controls. In patients with hepatobiliary disease the SGC was more frequently abnormal (83%) than values for serum albumin (7%), prothrombin time (17%), bilirubin (22%), alkaline phosphatase (45%), aspartate transaminase (57%) and gammaglutamyl transpeptidase (63%). The cumulative frequency of abnormality of these six tests was equal to tha...

1982-01-01

333

Modification of some biochemical parameters and of the concentration of some serum ions in Cyprinus carpio L. species grown under different sanitary-veterinary conditions  

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The paper discusses the modifications produced in some biochemical indices as well as in the concentration of serum ions, in fry carp, during the period of active feeding, in two experimental ponds of the Iasi district, providing different conditions of ichtyo-pathological prevention. The average values of glucose and alkaline phosphatase from the blood, as well as those of cholesterol, ureic nitrogen, Na+, Ca2+ and K+ from the serum, have been determined after 120 days of experiment (June - ...

2008-01-01

334

Effects of Aqueous Stem Bark Extract of Cissus populnea on Some Serum Enzymes in Normal and Alloxan Induced Diabetic Rats  

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This study was undertaken to assess the effect of 100 mg kg-1 body weight of aqueous stem bark extract of Cissus populnea on serum enzyme levels in normal and alloxan induced diabetic rats. The four weeks experimental protocol involving intragastric administration of the extract revealed a significant increase (P<0.05) in the level of serum alkaline and total acid phosphatase only as a result of diabetes induction. The treatment with Cissus has also revealed a sign...

Geidam, M. A.; Adoga, G. I.; Sanda, F. A.

2004-01-01

335

Correlation of Serum Magnesium with Serum Parathormone Levels in Patients on Regular Hemodialysis  

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Full Text Available Secondary hyperparathyroidism (SHPT is a common, important, and treatable complication of end-stage renal disease. This study was conducted to investigate the role of serum magnesium (Mg in regulating the secretion of parathyroid hormone (PTH by the parathyroid gland in patients on maintenance hemodialysis (HD. Pre-dialysis serum levels of calcium (Ca, phosphorus (P, Mg, alkaline phosphatase (ALP, intact serum PTH (iPTH, serum 25-hydroxy Vitamin D (25-OH Vit D and plasma bicarbonate (HCO3 were measured. The Urea Reduction Rate as well as duration and dosage of HD treatment were noted. Our study did not show any significant correlation between serum Mg levels and duration of HD treatment, levels of serum ALP, and plasma HCO3, Ca and P. An inverse correlation, albeit insignificant, was found between the serum Mg levels and iPTH (r=-0.30 p=0.079; also, a significant positive correlation was found between serum Mg levels and serum 25-OH Vit D levels (r= 0.40 p= 0.009. Our findings are in agreement with previous data, which suggest that factors other than serum Mg are more important in the regulation of PTH secretion in HD patients. A positive and strong association between serum Mg with 25-OH Vit D needs to be studied in greater detail.

Baradaran Azar

2006-01-01

336

Evaluation of four reagents for delipidation of serum  

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Four reagents, Aerosil 380, Freon 113, Dextran sulfate 500-S, and a mixed organic solvent were tested for their abilities to produce optically clear, pooled human serum. Aerosil-380, a silicon dioxide, removed 95% of serum cholesterol and triglycerides, and 80% of the free fatty acids. A mixed organic solvent (n-butanol:diisopropyl ether) was equally effective, but also removed nearly all endogenous alkaline phosphatase and lactate dehydrogenase. Freon-113 and Dextran sulfate 500-S removed about half of the serum cholesterol and triglycerides. The serum content of several non-lipid components was unaffected by Aerosil-380, Freon-113, and Dextran sulfate treatments; however, the mixed organic solvent removed 69% of the endogenous calcium. Light scattering data revealed that treatment with all reagents except the mixed organic solvent resulted in optically-clear serum products.

Agnese, S.T.; Spierto, F.W.; Hannon, W.H.

1983-04-01

337

Efecto del tratamiento con praziquantel sobre la actividad de la fosfatasa alcalina, fosfatasa acida, superoxido dismutasa en extractos crudos y productos de excreción-secreción de gusanos de Schistosoma mansoni / Effect of Treatment with Praziquantel on the activity of alkaline phosphatase acid phosphatase, superoxide dismutase in Crude Extracts and Excretion-secretion Products of Schistosoma mansoni worms.  

Scientific Electronic Library Online (English)

Full Text Available SciELO Venezuela | Language: Spanish Abstract in spanish Venezuela se encuentra entre los países sudamericanos afectados por la esquistosomiasis y la quimioterapia con praziquantel (PZQ) es la principal estrategia de control. Se determino el efecto cuantitativo del tratamiento con praziquantel sobre la actividad de la Fosfatasa Alcalina (ALP), Fosfatasa A [...] cida (ACP) y Superoxido Dismutasa (SOD), en antígenos solubles (ASG) y productos de excreción-secreción (PESG) de gusanos hembras y machos condición control (ASGHc, ASGMc, PESGHc and PESGMc) o incubados con PZQ in vitro (ASGMpzq, ASGHpzq, PESGMpzq and PESGHpzq). Las proteínas totales se determinaron por colorimetría, la SOD y ACP mediante espectrofotometría y la ALP por fluorometría. Se encontró una mayor concentración de proteínas en las ASG de gusanos no tratados, y en las preparaciones obtenidas luego de la incubación con PZQ in vitro, en los PESG, un incremento en la actividad ACP en los ASG y PESG preparados con gusanos no-tratados, y una disminución de dicha actividad en los ASG y PESG tratados. La SOD, evidenció en los ASG una disminución estadísticamente significativa en los gusanos tratados. La concentración de la ALP disminuyó significativamente en los ASG y PESGH de gusanos tratados en relación a los gusanos no tratados. En conclusión, se observó una disminución en las proteínas totales, actividades enzimáticas ACP y SOD, y concentración de ALP, en ASG y PESG obtenidos con gusanos tratados. Abstract in english Venezuela is among South American countries affected by schistosomiasis and chemotherapy with praziquantel (PZQ) is the main control strategy. We determined the quantitative effect of treatment with PZQ on alkaline phosphatase activity (ALP), acid phosphatase (ACP) and superoxide dismutase (SOD) in [...] soluble antigens of worms (SWAP) and excretion-secretion products (EEP) of male and female worms (SMWAPc, SFWAPc, ESPWMc and ESPWHc) or incubated with PZQ in vitro (SMWAP PZQ, SFWAP PZQ, ESPWM PZQ and ESPWH PZQ). Total proteins were determined by colorimetry, SOD and ACP by spectrophotometry and fluorometry ALP. There was higher protein concentration in the untreated worms EG, and the preparations obtained after incubation with PZQ in vitro, in the EG, an increase in ACP activity in the EG and PG prepared with non-treated worms and a decrease of such activity on the EG and treated PG. On the other hand, SOD activity, the EG showed statistical significance in the treated worms. In the PG showed the same behavior, but those differences were not statistically significant. Similarly, there was a decrease in the concentration of ALP noticeable in the EG and worm PGh treated worms relative to untreated statistically significant. In conclusion, we observed a decrease in total protein, ACP and SOD enzyme activities and concentration of ALP, and EG in PG treated worms.

Emilia E, Barrios; Jesús, Rodríguez; Naim, Richani; Wolfan, Araque; Juan F, Quintana; Lisset, Sánchez.

338

Chromogranin A as Serum Marker for Gastroenteropancreatic Neuroendocrine Tumors: A Single Center Experience and Literature Review  

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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...

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

2012-01-01

339

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  

Directory of Open Access Journals (Sweden)

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.

N.J.F. Oliveira

2005-06-01

340

Serum neuron-specific enolase (S-NSE) and the prognosis in small-cell lung cancer (SCLC): a combined multivariable analysis on data from nine centres.  

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The influence of pretreatment serum neuron-specific enolase (S-NSE) in addition to more conventional prognostic factors on survival duration in small-cell lung cancer (SCLC) was investigated in 770 patients from nine centres in six countries. The other variables included stage of disease, performance status (PS), age, sex, serum lactate dehydrogenase (S-LDH), serum alkaline phosphatase (S-AP), and serum carcinoembryonic antigen (S-CEA). Increased values of S-NSE (> 12.5 micrograms-1 l) were o...

1996-01-01

 
 
 
 
341

Serum and urinary measurements of prostatic acid phosphatase (PAP and prostatic specific antigen (PSA in dogs Mensurações sérica e urinária de fosfatase ácida prostática e antígeno prostático específico em cães  

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Full Text Available Serum and urinary prostatic acid phosphatase (PAP and prostatic specific antigen (PSA from 20 dogs were measured. PAP and PSA tests were carried out in authomatized equipment with commercial kits used for humans. Mean PAP serum value was 0.7U/l and urinary 0.1U/l. Mean serum and urinary PSA were 0.005ng/dl and 0.004ng/dl, respectively. In vivo determination of these two biomarkers in dogs is a new form of diagnosis in veterinary medicine and these values should be correlated with the morphological lesion of the prostate gland.Realizaram-se mensurações sérica e urinária de fosfatase ácida prostática (PAP e antígeno prostático específico (PSA de 20 cães. Os testes de PAP e PSA foram feitos em um equipamento automatizado, com o uso de kits comerciais para humanos. A média de PAP sérico foi de 0,7U/l e urinário 0,U/l. As médias do PSA sérico e urinário foram 0,005ng/dL e 0,004ng/dl, respectivamente. A determinação do dois biomarcadores in vivo é uma nova opção de diagnóstico na medicina veterinária e os valores obtidos devem ser correlacionados com a lesão morfológica da próstata.

R.L. Amorim

2004-06-01

342

THE POSSIBLE EFFECT OF SILDENAFIL CITRATE AND FENUGREEK SEED POWDER ON ENHANCING SERUM TESTOSTERONE LEVELS IN ADULT MALE ALBINO RATS  

International Nuclear Information System (INIS)

Sildenafil citrate is a phosphodiesterase 5 inhibitor (PDE5) that increases cyclic guanosine monophosphate (cGMP) which improves vasodilatation and there is a hypothesis that fenugreek seeds have 3 mechanisms by which it may enhance serum testosterone levels. The present study aimed to evaluate the effect of sildenafil citrate and fenugreek seeds alone or in combination on serum testosterone levels in normal adult male albino rats. Besides, total protein, albumin, globulin, bilirubin levels and alanine transaminase, aspartate transaminase and alkaline phosphatase activities were evaluated. The present study claimed that single or combined treatment with sildenafil and fenugreek seed powder (FSP) may enhance serum testosterone levels in adult male albino rats

2008-01-01

343

Alterations in certain Enzymatic Activities in Liver and Serum of Male Rats Treated with Endotoxin Or Exposed To Gamma Radiation  

International Nuclear Information System (INIS)

This study was performed to determine the effects of endotoxins and gamma radiation on glutamic oxalacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), acid phosphatase (ACP) and alkaline phosphatase (ALP). Endotoxin (isolated from Escherichia coli, Serotype 055) was supplemented to rats by gavage with a dose level of 20 mg/kg /day for a period of four weeks. Whole body gamma irradiation was carried out by exposure of rats to 4 Gy delivered as 0.5 Gy twice weekly. The results demonstrated that endotoxin administration as well as exposure to gamma radiation produced significant increases in the activities of liver transaminases and acid phosphatase enzymes 1,2 and 4 weeks post treatment. In the serum of endotoxin treated rats, the activities of transaminases showed non-significant changes while significant increases were observed in the serum of irradiated rats

2004-01-01

344

Dephosphorylation of Centrins by Protein Phosphatase 2C ? and ?  

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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 ...

2009-01-01

345

Dephosphorylation of Centrins by Protein Phosphatase 2C ? and ?  

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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 wa...

2009-01-01

346

Gamma radiation effects on liofilized human serum  

International Nuclear Information System (INIS)

Human freeze dried serum was artificially contaminated with Flavobacterium sp. for studying the effects of gamma radiation of it. The radiobiological parameters of the contaminator were determined and the sterilization dose was set. The quality of the product irradiated at both, calculated sterilization dose (8.5 kGy) an another one about 25 kGy was determined. It was made according to: sterility testing, total proteins, pH enzymes (alanina-aminotransferase, aspartato-aminotransferase, alkaline phosphatase), protein electrophoresis, fast performance liquid chromatographic and effect on the cellular growth. From the latter was concluded that the calculated sterilization dose was adequate form keeping the biological properties and viability of the irradiated serum. Nevertheless, the dose of 25 k Gy was not adequate because of its dangerous effects on the cell culture

1995-01-01

347

Production of two phosphatases by Lysobacter enzymogenes and purification and characterization of the extracellular enzyme.  

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Lysobacter enzymogenes produces an extracellular phosphatase (EC. 3.1.3.1) during the stationary phase of growth. The cells also produce a cell-associated alkaline phosphatase. This enzyme is found in the particulate fraction of cell extracts and may be membrane bound. The production of both phosphatases, especially the extracellular enzyme, is reduced by inorganic phosphate. The extracellular phosphatase was purified to a specific activity of 270 U/mg primarily by chromatography on carboxyme...

Von Tigerstrom, R. G.

1984-01-01

348

Investigations of serum HPL during pregnancy using two different radioimmunoassays  

International Nuclear Information System (INIS)

The interassay investigations showed that it is absolutely necessary to standardize the HPL antisera as well as the standard sera, as it is otherwise impossible to compare and interpret the findings of different HPL radioimmunoassays. The investigations have shown that in addition to conventional clinical examinations and laboratory test methods (urine estriol determination, DHEAS-dehydroepiandrosterone sulphate test-, urine pregnandiol determination, and determination of heat-resisting alkaline serum phosphatase), HPL concentration determination is a parameter of the nutritive function of the placenta. (orig.)

1978-01-01

349

Rapamycin selectively alters serum chemistry in diabetic mice  

Science.gov (United States)

The study was undertaken to explore the effect of rapamycin, an anti-inflammatory agent, on the metabolic profile of type 2 diabetic mice. Seven-month-old diabetic db/db mice and their lean littermate non-diabetic controls (db/m) were randomized to receive control chow or chow mixed with rapamycin (2.24 mg/kg/day) (each group n =20, males and females) for 4 months and sacrificed. Serum samples were analyzed for the measurement of glucose, creatinine, blood urea nitrogen (BUN), alkaline phosphatase (ALP), alanine aminotransferase (ALT), total cholesterol, total triglyceride, and total protein, using the automated dry chemistry analysis. Rapamycin elevated serum glucose in female diabetic mice. Serum creatinine tended to be higher in diabetic mice but was not affected by rapamycin; there was no difference in BUN levels among the groups. Serum ALP was elevated in diabetic mice and rapamycin lowered it only in female diabetic mice; serum ALT levels were increased in female diabetic mice, unaffected by rapamycin. Serum total protein was elevated in diabetic mice of both genders but was not affected by rapamycin. Diabetic mice from both genders had elevated serum cholesterol and triglycerides; rapamycin did not affect serum cholesterol but decreased serum total triglycerides in male diabetic mice. We conclude that rapamycin elicits complex metabolic responses in aging diabetic mice, worsening hyperglycemia in females but improving ALP in female diabetic and total triglycerides in male diabetic mice, respectively. The metabolic effects of rapamycin should be considered while performing studies with rapamycin in mice.

Tabatabai-Mir, Hooman; Sataranatarajan, Kavithalakshmi; Lee, Hak Joo; Bokov, Alex F.; Fernandez, Elizabeth; Diaz, Vivian; Choudhury, Goutam Ghosh; Richardson, Arlan; Kasinath, Balakuntalam S.

2012-01-01

350

Effects of Aqueous Stem Bark Extract of Cissus populnea on Some Serum Enzymes in Normal and Alloxan Induced Diabetic Rats  

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Full Text Available This study was undertaken to assess the effect of 100 mg kg-1 body weight of aqueous stem bark extract of Cissus populnea on serum enzyme levels in normal and alloxan induced diabetic rats. The four weeks experimental protocol involving intragastric administration of the extract revealed a significant increase (P<0.05 in the level of serum alkaline and total acid phosphatase only as a result of diabetes induction. The treatment with Cissus has also revealed a significant increase (P<0.05 in the level of serum acid phosphatase. However, there were no significant differences in the levels of aspartate and alanine aminotranferases as a result of both diabetes induction and Cissus populnea treatment.

M.A. Geidam

2004-01-01

351

The cell surface associated phosphatase activity of Mycobacterium bovis BCG is not regulated by environmental inorganic phosphate.  

Science.gov (United States)

Non-specific phosphomonoesterase activities (alkaline phosphatase (EC 3.1.3.1) and acid phosphatase (EC 3.1.3.2)) were examined at the cell surface of Mycobacterium bovis BCG. Using p-nitrophenylphosphate as the substrate, peaks of phosphatase activity were detected at pH 6.0, pH 10.0 and pH 12.0, suggesting the presence of one acid phosphatase and two alkaline phosphatases with distinct optimum pH values. Contrary to the situation observed in several other microorganisms, the expression of these enzymes is not regulated by the environmental inorganic phosphate concentration. PMID:11179639

Braibant, M; Content, J

2001-02-20

352

The effect of C3H mouse mammary tumour on the levels of serum and urine analytes in vivo.  

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A study of C3H mice implanted with mammary tumours has shown that the levels of serum total protein, alanine transaminase and alkaline phosphatase are all lower than those found in normal mice, while aspartate transaminase is higher. Serum urea values were similar to normal levels, but creatinine was lower in males and higher in females. In the male mice, urine protein and urine N-acetyl-beta-D-glucosaminidase (NAG) activity were lower than in normal mice. Comparisons were made with age and s...

1985-01-01

353

Detection of phosphatase activity in aquatic and terrestrial cyanobacterial strains  

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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 ?molpNP/s/dm3, was recorded in terrestrial strains compared to the studied water strains (1.11-5.96 ?molpNP/s/dm3. The activity of acid phosphatases was higher in most tested water strains (1.67-6.28 ?molpNP/s/dm3 compared to the activity of alkaline phosphatases (1.11-5.96 ?molpNP/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 Srbije, br. III 43002

Babi? Olivera B.

2013-01-01

354

Serum 25-hydroxyvitamin D and biochemical markers of bone metabolism in patients with juvenile idiopathic arthritis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Our objective was to evaluate the concentrations of serum 25-hydroxyvitamin D [25(OH)D], serum calcium, serum phosphorus, alkaline phosphatase, and parathormone (PTH) in patients with polyarticular juvenile idiopathic arthritis (JIA) and to associate them with disease duration and activity, bone min [...] eral density and use of medications. In a cross-sectional and controlled study, 30 patients with polyarticular JIA were evaluated and compared to 30 healthy individuals matched for age and gender. Clinical status, anthropometry, laboratory markers in both patients and controls, and bone mineral density, only in the patients, were measured. Of the 30 patients included in the study, 23 (76.7%) were female and 16 (53.3%) non-Caucasian; mean age was 14 years (range = 4 to 20 years). Mean disease duration was 5 years (range = 1 to 12 years). The mean concentrations of serum albumin-corrected calcium (9.04 ± 0.41?mg/dL) and alkaline phosphatase (153.3 ± 100.1 IU) were significantly lower in patients with JIA than in controls (P

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

2013-01-11

355

Clinical Significance of Detection of Serum TBA and ALP in Diagnosis of Intrahepatic Cholestasis of Pregnancy  

International Nuclear Information System (INIS)

To investigate the clinical value of serum total bile acid (TBA) and alkaline phosphatase (ALP) in diagnosis of intahrpatic cholestasis of pregnancy (ICP), the serum levels of TBA, ALP and cholyglycine (CG) in 47 cases with intahrpatic cholestasis of pregnancy and 60 normal pregnant women were tested by biochemistry analysis and radioimmunoassay. The results showed that the serum levels of TBA and ALP in patients with intahrpatic cholestasis of pregnancy were significantly higher than that of normal pregnancy women. There was a positively correlation between TBA and ALP with CG. The combined determination of serum TBA and ALP could be useful in the diagnosis of intahrpatic cholestasis of pregnancy. Automatic biochemistry analysis of TBA and ALP is more simple and rapid than CG detected by radioimmunoassay,and it is suitable for clinical laboratory application. (authors)

2009-06-01

356

Influence of dietary calcium on chlordecone-induced biochemical changes in serum of rat.  

Science.gov (United States)

Male, Sprague-Dawley rats were treated with different (1, 10, 50, and 100 ppm) concentrations of chlordecone (Cd) in calcium-sufficient (Ca-S) or calcium-deficient (Ca-D) diet for 15 days. No significant changes in serum total proteins were observed. However, serum nonprotein nitrogen compounds (urea, uric acid, and creatinine) and glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, creatine kinase, and alkaline phosphatase were significantly increased at 50 and 100 ppm of Cd. Chlordecone induced more increase in these serum components of rats fed with Ca-D as compared to Ca-S diet. Increased serum nonprotein nitrogen compounds and enzymes indicate Cd-altered glomerular and hepatic functions. PMID:7504618

Chetty, K N; Walker, J; Brown, K; Ivie, G W

1993-10-01

357

Correlation of Serum Parathormone With Hypertension in Chronic Renal Failure Patients Undergoing Hemodialysis  

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Full Text Available Purpose: To consider the effect of serum parathormone on severity of hypertension in end-stage renal failure patients undergoing hemodialysis treatment. Patients and Methods: A cross-sectional study was performed on patients with end-stage renal disease undergoing maintenance hemodialysis treatment. Serum calcium, phosphorus, alkalene phosphatase, serum albumin and intact PTH levels were measured. Stratification of hypertensive patients was performed from stage one to three. A stage of zero means the absence of hypertension. Stages of hypertension were measured before treatment and at the beginning of the first hemodialysis treatment. Results: The total number of patients was 73 (F=28 M=45, including 58 non-diabetic (F=22 M=36 and 15 diabetic hemodialysis patients (F=6 M=9. The mean age of patients was 46.5±16 years. The mean period of time that patients had spent on hemodialysis was 21.5±23.5 months. Serum iPTH of total patients was 309±349 pg/ml and serum alkaline phosphatase of total patients was 413±348 IU/L. There was a significant positive correlation between the stages of hypertension and serum iPTH levels (r=0.200; p=0.045. There was no significant correlation between the stages of hypertension and serum alkalene phosphatase levels (r=0.135; p=0.128. A significant positive correlation between stages of hypertension with Ca x P products of patients (r = 0.231; p=0.027 was also seen. Conclusion: The relationship between serum iPTH and severity of hypertension in this group requires further research on nontraditional causes of hypertension in hemodialysis patients. Hypertension and secondary hyperparathyroidism both interact in the process of accelerated atherosclerosis in hemodialysis patients. This combination may aggravate the rapid progressive athrosclerosis process

Baradaran, A.

2006-01-01

358

Do distinct water chemistry, reservoir age and disturbance make any difference on phosphatase activity?  

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Alkaline phosphatase activity was assessed concomitantly with total phosphorus, orthophosphate and phosphomonoester concentrations in two meso-eutrophic reservoirs with distinct age and subjected to different kinds of environmental influence. Differences in conductivity, temperature and pH were found. However, during the study period alkaline phosphatase activity was similar in both reservoirs. Water colour was higher in S. Serrada Reservoir. This fact can be related to (1) reservoir age (2) ...

Geraldes, Ana Maria; Boavida, Maria-jose?

2003-01-01

359

Chaperon-like Activation of Serum-Inducible Tryptophanyl-tRNA Synthetase Phosphorylation through Refolding as a Tool for Analysis of Clinical Samples1  

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Tryptophanyl-tRNA synthetase (TrpRS) expression alters in colorectal (CRC), pancreatic (PC), and cervical (CC) cancers. Here, phosphorylation of unfolded TrpRS and its fragments is stimulated by human cancer sera (CS; n = 13) and serum of rabbit tumor induced by Rous sarcoma virus, unaffected by donor sera (NS; 11/15) and abolished by alkaline phosphatase. At 20 years of follow-up, serum-inducible TrpRS phosphorylation found years before healthy donors (3/15) diagnosed with PC, CRC, or leukem...

Paley, Elena L.

2011-01-01

360

Serum Zinc Values in Adult Patients Undergoing Bone Marrow Transplantation  

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Full Text Available "nIntroduction: Zinc (Zn deficiency can cause significant defects in cellular immunity. Hematopoietic stem cell transplantation (HSCT patients usually experience serious deficiencies of all components of the immune system. Therefore, the maintenance of a normal Zn status may be important in this group of patients. "nPatients and Methods: Serum Zn levels were analyzed in 55 patients during the HSCT period. As Zn-related factors, serum copper (Cu levels and alkaline phosphatase (ALP activity were also measured. "nResults: There was decrease in Zn values immediate post-transplant period (at day +10 when compared to pre-HSCT levels (P=0.06. In patients who developed hypozincemia, adverse events appeared to occur more frequently. "nConclusion: This study suggests that maintaining a normal Zn status can be important in HSCT patients and Zn deficiency may be a risk factor causing adverse effects.

M Hadjibabaie

2009-07-01

 
 
 
 
361

Alkaline perturbation  

International Nuclear Information System (INIS)

This session gathers 4 articles dealing with: effect of deviation from equilibrium on dissolution rate of smectite under hyper-alkaline condition (T. Sato, Y. Otani, H. Takayama, S. Yokoyama, C. Oda, A. Honda, T. Yoneda); the influence of high pH fluid circulation on the mechanical behaviour of compacted clayey soil (O. Cuisinier, F. Masrouri, M. Pelletier, F. Villieras) the alteration of montmorillonites in saline solutions (H.J. Herbert, J. Kasbohm); and the organic matter-metals (Ti, Cr, Fe) interactions at the Khushaym Matruk natural analogue, Central Jordan (M. Elie, I. Techer, L. Trotignon, H. Khoury, E. Salameh, D. Vandamme, P. Boulvais, S. Fourcade)

2007-09-17

362

COMPARISON OF SERUM YST/ASAT, ALAT AND ALP LEVELS IN HEPATIC DISEASES  

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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.

DR. M ZAHRAIE

1987-06-01

363

Correlation of Serum Parathormone with Hypertension in Chronic Renal Failure Patients Treated with Hemodialysis  

International Nuclear Information System (INIS)

To consider the correlation of serum parathromone on severity of hypertension in end stage renal disease (ESRD) patients on hemodialysis (HD). A cross-sectional study was done on patients with ESRD on treatment with maintenance HD. Levels of serum calcium, phosphorous, alkaline phosphatase, albumin and intact parathormone (iPTH) were measured. Stratification of hypertensive patients was done from stage one to three. The total number of patients studied was 73 (Females=28, Males=45), consisting of 58 non-diabetic (F=22, M=36) and 15 diabetic patients (F=6, M=9). The mean age of the study patients was 46.5+-16 years. The mean duration on HD of the study patients was 21.5+-232.5 months. The mean serum PTH of the study patients was 309+-349 pg/ml and the mean serum alkaline phosphatase was 413+-348 IU/L. There was a significant positive correlation between the stage of hypertension and serum PTH levels (r=0.200, p=0.045). Also, there was a significant positive correlation between stage of hypertension and calcium-phosphorus product (r=0.231, p=0.027). There was no significant correlation between stage of hypertension and serum ALP (r=0.135, p=0.128). Relationship between serum PTH and severity of hypertension in patients on HD needs to be studied in more detail. Hypertension and secondary hyperparathyroidism interact in the process of accelerated atherosclerosis in HD patients thus warranting appropriate measures to control hyperparathyrodism vigorously. (author)

2005-01-01

364

Phosphatase activities in pesticide-treated growing and developing cells of Dictyostelium discoideum.  

Science.gov (United States)

The effects of benzene hexachloride (BHC), an organochlorine pesticide, on cell growth and phosphatase activities were studied in the growing and developing stages of cells of the cellular slime mould Dictyostelium discoideum exposed to BHC (containing alpha-, beta-, delta- and gamma-isomers) at concentrations of > or = 60 micrograms ml-1. A significant increase in acid and alkaline phosphatase activities was recorded in both growing and differentiating Dictyostelium cells treated with different doses (> or = 60 micrograms/ml) of BHC. The cytotoxicity of BHC has been correlated with the stimulation of acid and alkaline phosphatase activities of this organism. PMID:7690788

Gayatri, R; Chatterjee, S

1993-01-01

365

Changes in uterine protein secretion during luteal and follicular phases and detection of phosphatases during luteal phase of estrous cycle in buffaloes (Bubalus bubalis).  

Science.gov (United States)

Changes in uterine proteins during different reproductive states and their functional significance though known in other species have not been established in buffaloes. An attempt has been made to unravel the changes in composition of buffalo uterine secretion with growth and regression of corpora-lutea during early, mid and late luteal and follicular phase of estrous cycle using gel filtration and electrophoresis techniques. Also the phosphatases activities in luteal phase uterine secretions have been studied. Gel filtration chromatography analysis revealed a protein peak in void volume of the column, the intensity of which was more in all the luteal phase samples than follicular phase samples. Alkaline phosphatase was also found eluted in the void volume. The other three uterus-specific peaks (Peaks V-VII) were detected below 13.7 kd molecular weight. There were at least five peaks of acid phosphatases activity in chromatogram. Silver staining of SDS-PAGE gel detected as many as 40 protein bands in the uterine fluid of which nine proteins were glycoproteins. Molecular weight (MW) comparison revealed the major protein band at 66 kd which could be serum albumin. Comparison of uterine proteins with serum protein bands revealed a 93.5 kd glycoprotein in buffalo serum that did not appear in uterine fluid and at least 11 uterus-specific protein bands (506, 470, 241, 114, 49, 38, 33, 26, 19.2, 16, and 14.3 kd). The 38 and 19.2 kd bands were luteal-stage specific. Intense periodic acid Schiff's (PAS) stained bands in uterine proteins compared to serum indicated glycosylation process in endometrial epithelial cells. The study suggested that buffalo uterine secretion contained mainly serum and several uterus-specific proteins of which few were luteal phase specific. Further study on characterizing the unique or most abundant proteins and defining their role in uterine functions would help to address the cause of low reproduction rate in buffaloes. PMID:16213013

Chandra Roy, Sudhir; Uma Suganthi, R; Ghosh, Jyotirmoy

2006-04-15

366

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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-t [...] ransferase (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.

PRL, Silva; OC, Freitas Neto; AC, Laurentiz; OM, Junqueira; JJ, Fagliari.

367

A sensitive enzyme-catalytic nanogold-resonance scattering spectral assay for alkaline phosphate.  

Science.gov (United States)

In pH 8.9 Tris-HCl buffer solutions, alkaline phosphatase (ALP) catalyzed the hydrolysis of ascorbic acid 2-phosphate (AAP) substrate to form ascorbic acid. Then H(3)PO(4) was added to stop the enzymatic reaction and HAuCl(4) was used to react with ascorbic acid to generate gold nanoparticles that exhibited a resonance scattering (RS) peak at 600 nm. Under the selected conditions, when the activity of ALP increased, the formed ascorbic acid and gold nanoparticles also increased. Thus, the RS intensity at 600 nm enhanced linearly. The linear range was 0.06-22 U/L, with a detection limit of 0.03 U/L. The ALP in serum was analyzed, and the results were in agreement with those of the fluorescence method. PMID:22113359

Jiang, Zhiliang; Wu, Meng; Liu, Gaosan; Liang, Aihui

2012-06-01

368

Research on Phosphatases of Belladona Leaves and Their Purification  

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Full Text Available Through experimentation with several leaves it has been possible for us to point out the existance of two different acid phosphatases. We have studied in more detail the phosphatases of belldon a leaves (Atropa Belladona L. Solanacees. The great part of the phosphatase activity is water extractable. We have compared the activity of the soluble fraction with that not directly extractable by means of water. The insoluble fraction could not be solubilized in a satisfaetC'fY m.anner.The digestion by papaine produced a slight solubilizing effect; on the other hand salt solutions, neutral or alkaline, or water glycerol mixtures had no solubilizing effect on the enzyme, It has been possible to demonstrate the existence of two different phosphatases in the insoluble fraction: the first of the type II,

M. Khorsand

1957-01-01

369

Dephosphorylation of Centrins by Protein Phosphatase 2C ? and ?  

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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.

Marie-Christin Thissen

2009-01-01

370

Serum MDA, Antioxidant Vitamins and Erythrocytic Antioxidant Enzymes in Chronic Alcoholic Liver Disease – A Case Control Study  

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Full Text Available Objectives: The study aims to estimate the changes in the serum levels of lipid peroxidation product malondialdehyde (MDA, non-enzymatic antioxidants: vitamin A, E and C and erythrocyte enzymatic antioxidants: superoxide dismutase (SOD and catalase(CAT in chronic alcoholic liver disease. Background: Alcohol consumption accounts for about 50% of patients death from end stage liver disease in India. The increased free radical and their metabolites decrease the plasma antioxidants status in chronic alcoholic liver disease (CALD. Method: The study comprised of 100 healthy persons as controls and 100 diagnosed patients of chronic alcoholic liver disease as cases. The estimation of serum MDA, vitamin A, E, C and erythrocyte enzymatic antioxidants SOD and CAT, were carried out along with liver function parameters like serum aspartate amino transferase (AST, serum alanine aminotransferase (ALT, serum alkaline phosphatase (AP, serum gamma glutamyl transferase (GGT, serum total protein, serum albumin, prothrombin time (PT and serum bilirubin. Statistical analysis was done using unpaired “t” test. Result: The levels of serum MDA were significantly increased in patients with CALD (P<0.01 while antioxidants were significantly reduced as compared to controls (P<0.01. Conclusion: Increased levels of lipid peroxides and reduced antioxidants suggest that, oxidative stress plays a vital role in pathogenesis of chronic alcoholic liver disease

Sunita Pujar

2011-10-01

371

The relationship between the degree of liver fibrosis and serum markers in patient with hepatic diseases  

International Nuclear Information System (INIS)

To study the relationship between the degree of liver fibrosis and serum markers in patients with hepatic diseases, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and ?-glutamyl transpeptidase (GGT), total bilirubin (TBIL), albumin (ALB), globulin (GLO), platelet (PLT), prothrombin time (PT), procollagen type III (PIIINP), hyaluronic acid (HA), laminin (LN) and collagen type IV in 114 patients with different causes hepatic disease were determined. The liver puncture biopsy was also carried out to determine the stages of liver fibrosis. The results showed that the serum albumin, globulin, platelet, prothrombin time, PIIINP, HA, collagen type IV had significant difference in different stages of liver fibrosis. The serum platelets and albumin levels were negatively correlated with the degree of liver fibrosis. The serum PT and GLO levels were positively correlated with time course of liver fibrosis. The serum PIIINP, HA and collagen type IV levels were positively correlated with degree of liver fibrosis. The serum albumin, globulin, prothrombin time, platelets, PIIINP, HA, collagen type IV were correlated with the progress of the liver fibros