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1

Serum gamma glutamyl transferase and alkaline phosphatase in acute cholecystitis.  

UK PubMed Central (United Kingdom)

BACKGROUND: The serum level of gamma glutaryl transferase and alkaline phosphatase is raised in acute calculus cholecystitis and common bile duct stone. However, the rise in serum level of these enzymes in acute cholecystitis implies stone in the common bile duct is not well studied. Thus, it may lead to retained CBD stone on one side and unnecessary CBD exploration on the other during emergency laparoscopic cholecystectomy. The objective of the study is to predict presence of CBD stone by assessing serum level of gamma-glutamyltransferase (gamma-GT)and alkaline phosphatase. METHODS: A prospective study was designed which included 40 patients with clinically diagnosed and radiologically confirmed acute cholecystitis and 40 patients who had choledocholithiasis with or without cholangitis. Their serumgamma glutaryl transferase and alkaline phosphatase were analyzed. RESULTS: Both acute cholecystitis and CBD pathology had significant increase in alkaline phosphatase (p-value: 0.05). However, in acute cholecystitis there was 1.69±0.118 fold increase and in CBD pathology there was 2.5±0.57 fold increase in alkaline phosphatase than normal.(130 IU /L). There was no statistically significant difference ingamma- GT in both acute cholecystitis and CBD pathology(p-value: 0.390). However it increases by 2.8±0.47fold in acute cholecystitis and by 2.2±0.16 in CBD pathology(p value: 0.627). CONCLUSIONS: Although there is rise in serum?-GT and alkaline phosphatase level in acute cholecystitis and CBD stone,only more than 2.5 fold rise in serum alkaline phosphatase level predicts CBD stone.

Thapa PB; Maharjan DK; Suwal B; Byanjankar B; Singh DR

2010-10-01

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

DEFF Research Database (Denmark)

BACKGROUND: The bone mineral content of premature infants at term is lower than in mature infants at the same postconceptional age. Serum alkaline phosphatase and serum phosphate are often used as indicators of bone mineralisation. OBJECTIVE: To analyse the association between bone mineral content and serum alkaline phosphatase and serum phosphate. METHODS: Serum alkaline phosphatase and phosphate were measured at weekly intervals during admission in 108 premature infants of gestational age below 32 weeks (mean (SD) gestational age 29 (2) weeks; mean (SD) birth weight 1129 (279) g). Bone mineral content was measured at term (mean gestational age 41 weeks) by dual energy x ray absorptiometry and corrected for body size. RESULTS: Serum alkaline phosphatase was significantly negatively associated with serum phosphate (p < 0.001). Bone mineral content was not associated with mean serum alkaline phosphatase (p = 0.8), peak serum alkaline phosphatase (p = 0.5), or mean serum phosphate (p = 0.2) at term. CONCLUSION: Routine measurements of serum alkaline phosphatase and serum phosphate are of no use in predicting bone mineralisation outcome in premature infants.

Faerk, J; Peitersen, Birgit

2002-01-01

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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; Hou Lingling; Lu Na; Mohamed Asrah A; Wang Tao; Huang Yaling

2012-01-01

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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 (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; Junichi Shizuku; Takashi Ohba; Takashi Kabaya; Kosaku Nitta

2011-01-01

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Serum alkaline phosphatase screening for vitamin D deficiency states  

International Nuclear Information System (INIS)

Objective: To determine whether serum vitamin D levels are correlated with serum levels of alkaline phosphatase or not. Study Design: Cross-sectional, observational study. Place and Duration of Study: Multi-centre study, conducted at Liaquat National Hospital and Medical College, National Medical Centre and Medicare Hospital, Karachi, from January to October 2009. Methodology: Patients attending the Orthopaedic OPDs with complaints of pain in different body regions and serum vitamin D/sub 3/ levels of greater or equal to 30 ng/ml were included in the study. Patients with vitamin D deficiency were further categorized into mild deficiency or insufficiency (vit. D/sub 3/ = 20-29 ng/ml), moderate deficiency (vit. D/sub 3/ = 5 - 19 ng/ml) and severe deficiency forms (vit. D/sub 3/

2012-01-01

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Alkaline phosphatase bone isoenzyme activity in serum in various degrees of micromorphometrically assessed renal osteopathy.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase bone isoenzyme activity in serum correlates significantly to micromorphometrically assessed parameters of bone apposition as well as resorption. Though as both processes appear to be coupled in renal osteopathy, bone isoenzyme activity seems to reflect mainly hyperosteoid states. The detection of abnormal findings is impressively better by bone isoenzyme analysis as compared with total alkaline phosphatase activity; pathological values are found even at the initial phase of osteopathy in early renal failure. The determination of alkaline phosphatase bone isoenzyme activity in serum is a sensitive and reliable diagnostic tool in assessing the beginning and degree of metabolic bone disease.

Siede WH; Seiffert UB; Bundschuh F; Malluche HH; Schoeppe W

1980-06-01

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Outcome predictability of serum alkaline phosphatase in men with pre-dialysis CKD  

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Background. Serum alkaline phosphatase (ALP) increases in patients with chronic kidney disease (CKD) and high-turnover bone disease. ALP may represent an adjunct marker of high bone turnover devoid of drawbacks of serum parathyroid hormone (PTH), and it may also be associated with cardiovascular cal...

Kovesdy, Csaba P.; Ureche, Vitalie; Lu, Jun L.; Kalantar-Zadeh, Kamyar

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Relationship of serum and saliva calcium, phosphorus and alkaline phosphatase with dry mouth feeling in menopause.  

UK PubMed Central (United Kingdom)

OBJECTIVES: The aim of this study was to compare serum and saliva calcium, phosphorus and alkaline phosphatase of menopausal women with/without dry mouth (DM) feeling. BACKGROUND: The composition of saliva in menopause women with/without DM feeling is different. Some of these differences are in hormones that are related to bone turnover. METHODS: A case-control study was carried out on 60 selected menopausal women aged 45-79 years with or without DM feeling (30 as case, 30 as control), conducted at the Clinic of Oral Medicine, Tehran University of Medical Sciences. The phosphorus concentration was measured by photometrical measurement of the blue colour formed after the addition of ammonium molybdate and stannous chloride; calcium was measured by Arsenazo reaction; and alkaline phosphatase by the pNPP-AMP method. Statistical analysis of Student's t-test was used. RESULTS: The mean serum phosphorus and alkaline phosphatase, stimulated and unstimulated saliva calcium and alkaline phosphatase levels were significantly higher in the menopausal women suffering from DM. There were no significant differences between groups regarding saliva phosphorus and serum calcium concentration. CONCLUSION: Calcium, phosphorus and alkaline phosphatase appear associated with DM feeling in menopause.

Agha-Hosseini F; Mirzaii-Dizgah I; Moosavi MS

2012-06-01

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

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Full Text Available 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 concentration and alkaline phosphatase activity in children 1-15 years old in different growth and development period and of different sexes and to fortify the influence of growth and development dynamics on biohumoral status in healthy male and female children. Material and methods. We evaluated 117 healthy children of both sexes from 1-15 years of age and divided them into three age groups: 1-5, 6-10 and 11-15 years. We followed the serum calcium and phosphorus concentration and alkaline phosphatase activity in different groups and in different sexes. Results and conclusion. Our investigation found significantly higher values of serum calcium in boys than in girls with no important changes between the age groups and significantly higher values of serum phosphorus in the youngest age group in all children and in different sexes with no important sex differences. Alkaline phosphatase activity followed the growth spurt and was the biggest in 6-10 years group in girls and in 11-15 years group in boys.

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  

International Nuclear Information System (INIS)

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; Ghalaut Veena; Gupta Prem; Arora Puneet; Tandon S

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.

Nishimura, H.; Harada, T.; Nawata, J.; Hayakawa, M.; Nishioka, M.; Takemoto, T.; Yokoyama, T.; Takahashi, M.

1982-12-01

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

International Nuclear Information System (INIS)

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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Alkaline phosphatase, serum phosphate, and incident cardiovascular disease and total mortality in older men.  

UK PubMed Central (United Kingdom)

OBJECTIVE: We have examined the association between serum phosphate and alkaline phosphatase (ALP) with incident cardiovascular disease (CVD) outcomes and total mortality in older men. APPROACH AND RESULTS: A prospective study of 3381 men, aged 60 to 79 years, without a history of myocardial infarction or stroke followed up for an average 11 years during which there were 605 major CVD events (fatal coronary heart disease and nonfatal myocardial infarction, stroke, and CVD death) and 984 total deaths. ALP but not serum phosphate was associated with increased risk of coronary heart disease and overall CVD events which persisted after adjustment for CVD risk factors and markers of inflammation and after exclusion of men with chronic kidney disease (adjusted hazard ratio per SD, 1.19 [1.05, 1.34]; P=0.007 and 1.10 [1.01, 1.21]; P=0.04). In contrast, serum phosphate was only associated with increased CVD mortality owing to noncoronary heart disease or stroke causes (adjusted hazard ratio per SD, 1.35 [1.01, 1.83]; P=0.04). Both raised phosphate and ALP were associated with significantly increased total mortality after full adjustment and exclusion of men with chronic kidney disease. CONCLUSIONS: ALP but not serum phosphate is associated with coronary heart disease risk in elderly men. High levels of ALP and serum phosphate are both associated with increased total mortality.

Wannamethee SG; Sattar N; Papcosta O; Lennon L; Whincup PH

2013-05-01

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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; M.I. Qureshi; M.M. Jafari

2007-01-01

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Serum alkaline phosphatase isoenzymes in SD rats detected by polyacrylamide-gel disk electrophoresis.  

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Serum alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. In the present study, we assessed the usefulness of a commercially available polyacrylamide-gel disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co. Ltd., Tokyo, Japan) for identifying serum ALP isoenzymes in rats of the Sprague-Dawley strain (SD rats), which are commonly used in toxicity studies. We also examined age-related changes in serum ALP isoenzymes in SD rats. In order to identify the origin of each ALP isoenzyme, tissue ALP extracts from the liver, bone and small intestine (SI) and serum samples were treated with neuraminidase, antiintestinal ALP antibody, ALP inhibitor levamisole, and/or wheat germ agglutinin. It became clear that pretreatment of serum with neuraminidase is necessary for rat serum ALP isoenzyme analysis. The kit revealed that the main serum ALP isoenzymes in fasted 8-week-old intact rats were bone- and SI-derived and they tended to decrease with age. Serum liver-derived isoenzyme was slightly detected in both sexes of all ages examined, but it greatly increased in cholestasis model rats with bile-duct ligation, and rats of this model also had large molecular ALP detected in the stacking gel, suggesting hepatic damage. High-molecular intestinal ALP isoenzyme was slightly observed at the most cathodal side of the resolving gel. These results suggest that the present method is a useful tool for detecting serum ALP isoenzymes in SD rats and that concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity. PMID:22500783

Hatayama, Kazuhisa; Ichikawa, Yuko; Nishihara, Yoshito; Goto, Ken; Nakamura, Daichi; Wakita, Atsushi; Kobayashi, Jyunichi

2012-05-01

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Extremely elevated activity of serum alkaline phosphatase in gestational diabetes: a case report.  

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We report a case of a 25-year-old pregnant woman with gestational diabetes and increased activity (25-fold) of placental isozyme of alkaline phosphatase. Abdominal ultrasonographic scan revealed no hepatobiliary disease. After delivery, the alkaline phosphatase level decreased but did not return to the reference range. Similar abnormalities were found in the patient's first-degree relatives, which included a mother and a sister. PMID:14981410

Wojcicka-Bentyn, Janina; Czajkowski, Krzysztof; Sienko, Jacek; Grymowicz, Monika; Bros, Magdalena

2004-02-01

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Association between absolute tumor burden and serum bone-specific alkaline phosphatase in canine appendicular osteosarcoma.  

UK PubMed Central (United Kingdom)

BACKGROUND: In dogs with appendicular osteosarcoma (OSA), increased pretreatment serum bone-specific alkaline phosphatase (BALP) activity is a negative prognostic factor, associated with shorter disease-free intervals and survival times, but a biologic basis for observed differential serum BALP activities in canine OSA patients remains incompletely defined. OBJECTIVE: Serum BALP activity will correlate with absolute tumor burden in dogs with OSA. ANIMALS: This study included 96 client-owned dogs with appendicular OSA. METHODS: In canine OSA cell lines, the expression and membranous release of BALP was evaluated in vitro. The correlation between serum BALP activity and radiographic primary tumor size was evaluated in OSA-bearing dogs. In dogs developing visceral OSA metastases, serial changes in serum BALP activities were evaluated in relation to progression of macroscopic metastases, and visceral metastatic OSA cells were evaluated for BALP expression. RESULTS: In vitro, BALP expression was not associated with either tumorigenic or metastatic phenotype, rather the quantity of membranous BALP released was proportional with cell density. In dogs devoid of macroscopic metastases, there was a positive correlation between serum BALP activity and absolute primary tumor size. In dogs with progressive OSA metastases, serum BALP activity increased and coincided with the development of macroscopic metastases. OSA cells derived from visceral metastatic lesions retained BALP expression. CONCLUSIONS AND CLINICAL IMPORTANCE: Tumor burden is a determinant of serum BALP activity in dogs with appendicular OSA. The association between increased pretreatment BALP activity and negative clinical prognosis may simply be attributed to greater initial tumor burden, and consequently more advanced tumor stage.

Sternberg RA; Pondenis HC; Yang X; Mitchell MA; O'Brien RT; Garrett LD; Helferich WG; Hoffmann WE; Fan TM

2013-07-01

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Correlation of serum alkaline phosphatase with clinicopathological characteristics of patients with oesophageal cancer.  

Science.gov (United States)

Oesophageal cancer is endemic in some regions of the Islamic Republic of Iran and efforts have made to find factors that play a role in its prognosis. We retrospectively examined the correlation of serum alkaline phosphatase (ALP) levels with several clinicopathological characteristics of 207 cases of oesophageal carcinoma. The mean ALP level in patients with lymph node involvement was significantly higher [141 (SD 77) U/L] than with node negative cancers [116 (SD 63) U/L]. Patients with ALP levels 165 U/L were 3.29 times more likely to have lymph node involvement than patients with ALP levels < or = 165 U/L. There was no statistically significant correlation between ALP level and sex, age, tumour histological type, site and size of tumour, depth of penetration, distant metastasis, degree of differentiation, presence of lymphatic invasion and presence of simultaneous multiple cancers. Elevated ALP in patients with oesophageal cancer may predict lymph node involvement. PMID:22276495

Aminian, A; Karimian, F; Mirsharifi, R; Alibakhshi, A; Hasani, S M; Dashti, H; Jahangiri, Y; Ghaderi, H; Meysamie, A

2011-11-01

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ALP (Alkaline Phosphatase) Test  

Science.gov (United States)

... 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 ...

 
 
 
 
21

Isolated increase in serum alkaline phosphatase after liver transplantation: risk factors and outcomes analysis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Isolated increase in serum alkaline phosphatase (IISAlp) is frequently observed in liver transplant recipients visiting outpatient clinics. However, whether the increase is associated with risk factors or poor survival is unknown. METHODS: We retrospectively reviewed the medical records of liver transplant recipients who were followed up during 1999-2009 and had IISAlp 1 month after liver transplantation, which was sustained for at least 6 months. Clinical parameters, survival, and risk factors were analyzed and compared between recipients who survived longer than 6 months after transplantation. RESULTS: Among 307 liver transplant recipients, 44 had IISAlp. Compared with the control group, the patients with IISAlp were more frequently of the pediatric population, recipients of female donor or living-related partial liver grafts, and found to have biliary-related pretransplant disorders, lower body weight, and shorter warm ischemic time (P < 0.01). One patient with IISAlp died of acute myeloid leukemia during the follow-up period. The mean time to observation of IISAlp after liver transplantation was 6.3 ± 0.8 months. The mean follow-up duration was 5.5 ± 0.2 years. Stepwise multivariate analysis showed that being a pediatric or living-related liver transplant recipient was an independent risk factor for IISAlp, with adjusted hazard ratios (95% confidence interval) of 5.41 (2.59-11.28) and 3.0 (0.98-9.27), respectively. CONCLUSIONS: Therefore, being a pediatric or living-related liver transplant recipient was an independent risk factor for IISAlp. However, IISAlp was not associated with poor survival after liver transplantation. Hence, patients who have undergone liver transplantation do not require frequent routine examination of serum alkaline phosphatase levels.

Ho CM; Ho MC; Shau WY; Hu RH; Lai HS; Wu YM; Lee PH

2013-01-01

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Associations between serum bone-specific alkaline phosphatase activity, biochemical parameters, and functional polymorphisms of the tissue-nonspecific alkaline phosphatase gene in a Japanese population.  

UK PubMed Central (United Kingdom)

INTRODUCTION: We had demonstrated that single nucleotide polymorphism (787T>C) in the tissue-nonspecific ALP (TNSALP) gene was associated with the bone mineral density (BMD). BMD was the lowest among TNSALP 787T homozygotes (TT-type) and highest among TNSALP 787T>C homozygotes (CC-type) in postmenopausal women. In the present study, we investigated the effects of the TNSALP genotype on associations among serum bonespecific alkaline phosphatase (BAP), serum calcium, and phosphorus in healthy young Japanese subjects. METHODS: Young healthy adult subjects (n=193) were genotyped for the polymorphism, and we measured the levels of serum BAP, serum calcium, and phosphorus. Dietary nutrient intakes were calculated based on 3-day food records before the day of blood examinations. RESULTS: Grouped by the TNSALP genotype, a significant negative correlation between serum BAP and phosphorus was observed in 787T>C homozygotes (CC-type), but not in heterozygotes (TCtype), nor in 787T homozygotes (TT-type). CONCLUSIONS: In the present study, we revealed that the single nucleotide polymorphism 787T>C in the TNSALP gene had effects on the correlation between serum BAP and phosphorus in young adult subjects. These results suggest that variation in TNSALP may be an important determinant of phosphate metabolism. Our data may be useful for planning strategies to prevent osteoporosis.

Sogabe N; Tanabe R; Haraikawa M; Maruoka Y; Orimo H; Hosoi T; Goseki-Sone M

2013-01-01

23

[Effect of varying alloxan concentration on serum levels of glucose, calcium and alkaline phosphatase in the rat  

UK PubMed Central (United Kingdom)

Two groups of rats were injected intravenously 20 and 40 mg alloxan/kg of body weight respectively. It was shown that different doses of alloxan induced the increase of glucose concentration, activity of alkaline phosphatase and decrease of calcium level in the rat serum. The above changes more expressed in the groups of rats that received the higher dose of alloxan.

Stefanov MV

1993-07-01

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The effect of growth hormone treatment on serum bone alkaline phosphatase in growth hormone deficient children  

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Full Text Available Introduction: Growth hormone (GH) promotes longitudinal growth in children with growth hormone deficiency. Serum bone alkaline phosphatase (BALP) has been considered to be a good marker of bone formation with narrow circadian variation. The aim of the study was to determine the effect of GH treatment on serum BALP levels in children with idiopathic GH deficiency. Material and methods: A total of 26 prepubertal children (10 girls) with GH deficiency in mean age 9.06±1.24 yrs were enrolled in the study. They were treated with daily sc injections of GH (0.5 IU/kg/week). Serum BALP levels were measured using enzyme immunoassay (Alkaphase-B kit, Metra Biosystems) at baseline, 3 and 6 months of GH treatment. Results: A significant increase in serum BALP levels after 3 months of GH treatment was observed. No significant differences between BALP levels after 3 and 6 months of GH therapy were stated. Baseline serum BALP levels correlated positively with the height velocity during the 3 months period before the GH treatment initiation (r=0.63, p?0.05). The BALP increase after 3 months of treatment correlated with height velocity in that period (r=0.69, p?0.05). The change in BALP after 3 months of GH treatment correlated with the improvement in the height after 3 and 6 months (r=0.76, p?0.05) as well as after 12 months of GH treatment (r=0.77, p?0.05). Conclusions: We conclude that serum BALP appears to be a good early predictor of the growth responses to GH therapy in GH-deficient children.

Maria Korpal-Szczyrska; Anna Balcerska

2008-01-01

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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; E.A.C Nwanze

2004-01-01

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Persistent limb pain and raised serum alkaline phosphatase the earliest markers of subclinical hypovitaminosis D in Kashmir.  

Science.gov (United States)

The present study was an attempt to assess the cause of persistent pain in lower limbs among children from Kashmir. The study was conducted on one hundred children attending Paediatric out-patient department of Sher-i-Kashmir Institute of Medical Sciences, Srinagar. All the children were in the age group of 5 to 14 years. They showed markedly raised levels of serum alkaline phosphatase, whereas serum phosphorus, serum calcium levels and antistreptolycin O-titres were normal in 93% cases. None of them had any rheumatic or rheumatoid pathology. Among 15 suspected clinical rickets only three were established radiologically. Dietary and socio-economic history revealed deficient vitamin D intake and less exposure to sun. It was hypothesized that sub-clinical vitamin D deficiency could be a major cause of persistent pain in lower limbs and raised serum alkaline phosphatase could be the earliest marker of vitamin D deficiency. It was confirmed by injecting single dose of vitamin D (3 lac I. U.) which relieved bone pain and lowered the levels of serum alkaline phosphatase to normal within 14 weeks of initiation of therapy. PMID:2620972

Masood, H; Narang, A P; Bhat, I A; Shah, G N

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Persistent limb pain and raised serum alkaline phosphatase the earliest markers of subclinical hypovitaminosis D in Kashmir.  

UK PubMed Central (United Kingdom)

The present study was an attempt to assess the cause of persistent pain in lower limbs among children from Kashmir. The study was conducted on one hundred children attending Paediatric out-patient department of Sher-i-Kashmir Institute of Medical Sciences, Srinagar. All the children were in the age group of 5 to 14 years. They showed markedly raised levels of serum alkaline phosphatase, whereas serum phosphorus, serum calcium levels and antistreptolycin O-titres were normal in 93% cases. None of them had any rheumatic or rheumatoid pathology. Among 15 suspected clinical rickets only three were established radiologically. Dietary and socio-economic history revealed deficient vitamin D intake and less exposure to sun. It was hypothesized that sub-clinical vitamin D deficiency could be a major cause of persistent pain in lower limbs and raised serum alkaline phosphatase could be the earliest marker of vitamin D deficiency. It was confirmed by injecting single dose of vitamin D (3 lac I. U.) which relieved bone pain and lowered the levels of serum alkaline phosphatase to normal within 14 weeks of initiation of therapy.

Masood H; Narang AP; Bhat IA; Shah GN

1989-10-01

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

Science.gov (United States)

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

2013-01-10

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Serum Alkaline Phosphatase and Phosphate in Cerebral Atherosclerosis and Functional Outcomes After Cerebral Infarction.  

UK PubMed Central (United Kingdom)

BACKGROUND AND PURPOSE: Higher serum alkaline phosphatase (ALP) and phosphate levels are associated with atherosclerotic disease and an increased risk of cardiovascular events. However, the association of ALP/phosphate with cerebral atherosclerosis and prognosis in patients with acute stroke is not well known. METHODS: In 1034 patients with first-ever acute cerebral infarction, levels of ALP and phosphate were compared with (1) cerebral atherosclerosis and (2) poor long-term functional outcomes as defined by the modified Rankin Scale >2 at 3 months after stroke onset. RESULTS: ALP levels were not associated with cerebral atherosclerosis. However, higher levels of ALP were associated with a poor functional outcome (adjusted odds ratio per 1 SD, 1.25; 95% confidence interval, 1.04-1.50). Phosphate was associated with neither cerebral atherosclerosis nor functional outcome. CONCLUSIONS: A higher level of ALP was not associated with cerebral atherosclerosis but was an independent prognostic factor for long-term functional outcome after acute cerebral infarction.

Kim J; Song TJ; Song D; Lee HS; Nam CM; Nam HS; Kim YD; Heo JH

2013-09-01

30

Serum Alkaline Phosphatase Levels in Healthy Children and Evaluation of Alkaline Phosphatasez-scores in Different Types of Rickets  

Science.gov (United States)

Objective: Serum alkaline phosphatase (ALP) levels show great variation with age and sex in children and adolescents. Additionally, different buffers used even in the same method cause variable results. This detail is not usually taken into account in the evaluation. We aimed to study pediatric age- and sex-specific reference ranges for ALP by colorimetric assay using p-nitrophenyl phosphate as substrate and diethanolamine as buffer and also to compare the ALP levels in patients with different types of rickets. Methods: 1741 healthy children and adolescents (904 girls) were included in the study for normative data. 77 different ALP measurements from 38 nutritional rickets (NR), 7 vitamin D-dependent rickets (VDDR) and 8 hypophosphatemic rickets (HR) patients were included. Results: Reference values for ALP were constructed. ALP levels demonstrated a tetraphasic course with two peaks at infancy and puberty. There was no difference in ALP levels between boys and girls until puberty. However, higher ALP levels were noted at 10-11 years in girls (p=0.02) and at 12-13, 14-15, 16-17 years in boys (p<0.001). ALP levels start to decline after age 12 and 14 in girls and boys, respectively. Serum ALP levels were highest in the VDDR group and lowest in the HR group (median z-score values in HR, VDDR and NR were 3.6, 10.4 and 6.5, respectively; p<0.001). Similarly, plasma parathormone(PTH) levels ranged from highest to lowest in the VDDR, NR and HR groups (median values: 525, 237 and 98 pg/mL, respectively; p<0.001). Conclusions: This normative data will provide a basis for better evaluation of ALP levels determined by the described method. Furthermore, use of z-scores gives a more precise assessment of changes in ALP levels in rickets and other bone disorders. Conflict of interest:None declared.

Topcu, Burcu; Gokce, Ibrahim; Guran, Tulay; Atay, Zeynep; Omar, Anjumanara; Akcay, Teoman; Bereket, Abdullah

2011-01-01

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

International Nuclear Information System (INIS)

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

1993-01-01

32

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

Energy Technology Data Exchange (ETDEWEB)

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).

Garnero, P.; Delmas, P.D.

1993-10-01

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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...

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

34

Alkaline phosphatase activity in blood serum of dogs exposed to a mixture of external ?-radiation and internal ?-radiation  

International Nuclear Information System (INIS)

[en] Alkaline phosphatase activity in dog blood serum was studied for two years following separate and combined exposure to gamma radiation (6.45 to 51.6 mc/kg) and inhaled submicron 239Pu oxide containing 25% 241Am in chronically effective amounts (approx. 7-10 kBq/kg). Alkaline phosphatase activity was of an ondulatory nature and the significance of changes depended on the kind and the level of radiation as well as the time lapsed from the start of the expose. With the combined exposure to gamma and alpha radiation in the doses used no enhancement of the effec twaas noted as compared with the action of each factor applied separately

1988-01-01

35

Canine osteosarcoma cell lines from patients with differing serum alkaline phosphatase concentrations display no behavioural differences in vitro.  

UK PubMed Central (United Kingdom)

Osteosarcoma is an aggressive malignancy and represents the most frequent primary bone malignancy of dogs and humans. Prognostic factors reported for osteosarcoma include tumour size, presence of metastatic disease and serum alkaline phosphatase (ALP) concentration at the time of diagnosis. To date, there have been no studies to determine whether the behaviour of osteosarcoma cells differ based on serum ALP concentration. Here, we report on the generation of six canine osteosarcoma cell lines from osteosarcoma-bearing dogs with differences in serum ALP concentration. To determine whether in vitro behaviour differs between primary osteosarcoma cell lines generated from patients with normal or increased serum ALP, assays were performed to evaluate proliferation, migration, invasion and chemosensitivity. There were no significant differences in cell proliferation, migration, invasion or chemosensitivity between cell lines associated with normal or increased serum ALP concentration.

Holmes KE; Thompson V; Piskun CM; Kohnken RA; Huelsmeyer MK; Fan TM; Stein TJ

2013-03-01

36

[Changes in the polymorphism of the blood serum alkaline phosphatase in chickens undergoing subacute lindane treatment  

UK PubMed Central (United Kingdom)

Investigated were a total of 123 birds of the White Leghorn breed and 50 birds of the White Plymouth Rock breed. Preliminary studies were carried out on the LD50 amount of lindane (151 mg/kg body mass) with 6 groups of chickens. The birds were divided into 4 subgroups for each breed. The first subgroup was a control one and was not given lindane, and the remaining subgroups were offered the following amounts of lindane in the course of 5 successive days: II - 1/20 LD50 (7.6 mg/kg daily); III - 1/10 LD50 (15.1 mg/kg); and IV - 1/5 LD50 (30.2 mg/kg). Each single one of the test birds was treated individually by means of a rubber tube and a syringe with a pure substance of lindane dissolved in oil. The polymorphism of alkaline phosphatase in the blood plasms of chickens was determined through horizontal electrophoresis in starch gel after the method of Gahne [14]. It was found that at subacute treatment of the birds a drop off was observed of the S1 fraction, and, as a result, the frequence of the homozygotic alkaline-phosphatase phenotypes FF and SS rose, while that of FS1 and SS1 dropped. Subacute treatment with part of the birds led to the appearance of an additional fraction F1, which accompanied the SS, SS1, and SS2 phenotypes only. The changes that took place as a result of the subacute treatment were only phenotypic ones as the F1 and S1 fractions had no hereditary character.

Iotsov S; Iotova I; Atanasov K; Sotirov L

1983-01-01

37

Modulators of intestinal alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Small molecule modulators of phosphatases can lead to clinically useful drugs and serve as invaluable tools to study functional roles of various phosphatases in vivo. Here, we describe lead discovery strategies for identification of inhibitors and activators of intestinal alkaline phosphatases. To identify isozyme-selective inhibitors and activators of the human and mouse intestinal alkaline phosphatases, ultrahigh throughput chemiluminescent assays, utilizing CDP-Star as a substrate, were developed for murine intestinal alkaline phosphatase (mIAP), human intestinal alkaline phosphatase (hIAP), human placental alkaline phosphatase (PLAP), and human tissue-nonspecific alkaline phosphatase (TNAP) isozymes. Using these 1,536-well assays, concurrent HTS screens of the MLSMR library of 323,000 compounds were conducted for human and mouse IAP isozymes monitoring both inhibition and activation. This parallel screening approach led to identification of a novel inhibitory scaffold selective for murine intestinal alkaline phosphatase. SAR efforts based on parallel testing of analogs against different AP isozymes generated a potent inhibitor of the murine IAP with IC50 of 540 nM, at least 65-fold selectivity against human TNAP, and >185 selectivity against human PLAP.

Bobkova EV; Kiffer-Moreira T; Sergienko EA

2013-01-01

38

A retrospective study on craniofacial fibrous dysplasia: Preoperative serum alkaline phosphatase as a prognostic marker?  

UK PubMed Central (United Kingdom)

BACKGROUND: Craniofacial fibrous dysplasia (CFD) often requires surgery to correct facial deformity and prevent functional impairment. However, recurrence is common, and there is no reliable prognostic biomarker. The aim of this paper is to evaluate the possibility of using preoperative alkaline phosphatase (ALP) as a prognostic marker for CFD. MATERIAL AND METHODS: Forty-nine patients with CFD who underwent surgery from 2000 to 2011 were selected. The relationship between preoperative ALP and age, gender, lesion type and prognosis was investigated. RESULTS: The recurrence rate was 31.8% in patients who received conservative bone contouring. Patients with recurrence did not show significantly higher levels and abnormal rates of ALP than patients without recurrence. Young patients and those with polyostotic CFD showed higher ALP levels than adults and those with monostotic CPD (P < 0.05). Although CFD patients showed higher levels and abnormal rates of ALP than the control group, significant levels were not reached (P > 0.05). No correlation between age, gender, type, ALP and recurrence could be established using the logistic regression model. CONCLUSION: Preoperative ALP may not be a reliable prognostic marker of CFD based on the findings in this study. Close follow-up is recommended after conservative bone contouring.

Ma J; Liang L; Gu B; Zhang H; Wen W; Liu H

2013-10-01

39

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

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Full Text Available 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.Methods: Sixty-three neonates in the NICU were enrolled in this study. The neonates were divided into infected group (n=33) and non-infected group (n=30). The scores for neonatal acute physiology-perinatal extension II (SNAPPE-II) were calculated and interleukin-6 (IL-6), procalcitonin (PCT), BALP, OC and CTX were measured among the neonates with or without infections, and among the infected neonates before and after treatment.Results: The serum BALP levels were lower in the infected group than that in the non-infected group (p<0.01). The serum BALP levels increased markedly in the infected neonates after treatment (p<0.01). The serum BALP levels were also inversely correlated with SNAPPE-II of infected neonates before and after treatment (r=-0.56, p<0.05; r=-0.37, p<0.05, respectively). In infected neonates, the differences between serum BALP levels before and after treatment were inversely correlated with those of IL-6 levels (p<0.05). There were no significant changes in the OC, CTX and PCT levels in the infected or non-infected group before and after treatment.Conclusion: Our data suggest that serum BALP level might be used as a marker for assessing the severity of illness in infected neonates.

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

2012-01-01

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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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Full Text Available 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 the level of Ca was in the"nnormal range. Phosphor (P) was in the normal range in all patients over 17 years old and 80% under 17"nyears. The level of ALK in 75.8 percent of the patients over 17 years and 90% under 17 years was in the"nnormal range."nIn conclusion, no relationship was found between serum changes and P.G.C.G. It means P.G.C.G can be"ncompeletly independent lesions from serum changes.

Saheb-Jamei M;  Abdossamadi HR

2000-01-01

 
 
 
 
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ESTIMATION OF SERUM ALKALINE PHOSPHATASE, CHOLESTEROL, CALCIUM AND PHOSPHORUS DURING PRE-LA YING AND LAYING CONDITIONS IN DIFFERENT STRAINS OF CHICKENS  

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Full Text Available In order to estimate serum alkaline phosphatase, cholesterol, calcium and phosphorus during pre-laying and laying reproductive conditions, 60 hens of Desi, Fayoumi, Cross (Rhode Island Red X Fayoumi} and Nick Chick strains were maintained for one year. Five random blood samples from each strain were collected and analyzed during both pre egg laying and egg laying physiological conditions. It was observed that alkaline phosphatase activity increased significantly (P<0.05) during laying condition. Serum cholesterol remained unaffected by both the strain difference and laying condition. Serum calcium and phosphorus levels increased (P<0.05) during laying condition. The interaction of strain and stage of laying condition was found to exert significant (P<0.05) effect on serum calcium levels. The study showed that the availability of calcium and phosphorus in requisite quantities be provided in diet of laying hens to ensure sustained and quality egg production.

Bashir Mahmood Bhatti, Tanzeela Talat and Rozina Sardar

2002-01-01

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Effect of One Period of Aerobic Exercise on Serum Levels of Alkaline Phosphatase and Osteocalcin in Patients with Type 2 Diabetes  

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Full Text Available Introduction: The purpose of this study was to investigate the effects of a ten week period of aerobic exercise training on serum alkaline phosphates and osteocalsin in type 2 diabetic patients. Methods: In a quasi-experimental trial study twenty one male patients with type 2 diabetes(40-50 years) were randomly divided into exercise(n=11) and control(n=10) groups. The exercise group underwent a 10-week aerobic exercise program(three sessions per week, 45-60 minutes each session, at 50-65% of heart rate reserve). VO2max, BMI, fasting blood glucose and serum insulin, alkaline phosphatase and osteocalcin were measured at baseline and after exercise program. Results: Exercise program resulted in a significant increase in VO2max and a significant decrease in BMI, fasting blood glucose and HbA1c in exercise group; however, no significant changes were found in the insulin concentration, alkaline phosphatase and osteocalcin. The bone formation markers and other measured variables did not show significant change in control group. Conclusion: These results suggest that aerobic exercise leads to glycemic improvement in type 2 diabetic patients, but does not affect serum levels of alkaline phosphatase and osteocalcin.

Khorshidi; Matinhomaee; Azarbayjani; Hossein-nezhad

2011-01-01

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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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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 us...

Aliyu I; Isah H; Afonja O

44

Serum Proteins and Alkaline Phosphatase Levels in Patients with Tuberous Sclerosis  

Science.gov (United States)

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

Fischer, M. H.; And Others

1974-01-01

45

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 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; Bardapurkar Jayashree; Jain Anuradha

2009-01-01

46

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

UK PubMed Central (United Kingdom)

INTRODUCTION: Laboratory and clinical data have implicated endotoxin as an important factor in the inflammatory response to cardiopulmonary bypass. We assessed the effects of the administration of bovine intestinal alkaline phosphatase (bIAP), an endotoxin detoxifier, on alkaline phosphatase levels in patients undergoing coronary artery bypass grafting. METHODS: A total of 63 patients undergoing coronary artery bypass grafting were enrolled and prospectively randomized. Bovine intestinal alkaline phosphatase (n=32) or placebo (n=31) was administered as an intravenous bolus followed by continuous infusion for 36 hours. The primary endpoint was to evaluate alkaline phosphatase levels in both groups and to find out if administration of bIAP to patients undergoing CABG would lead to endogenous alkaline phosphatase release. RESULTS: No significant adverse effects were identified in either group. In all the 32 patients of the bIAP-treated group, we found an initial rise of plasma alkaline phosphatase levels due to bolus administration (464.27±176.17 IU/L). A significant increase of plasma alkaline phosphatase at 4-6 hours postoperatively was observed (354.97±95.00 IU/L) as well. Using LHA inhibition, it was shown that this second peak was caused by the generation of tissue non specific alkaline phosphatase (TNSALP-type alkaline phosphatase). CONCLUSIONS: Intravenous bolus administration plus 8 hours continuous infusion of alkaline phosphatase in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass results in endogenous alkaline phosphatase release. This endogenous alkaline phosphatase may play a role in the immune defense system.

Kats S; Brands R; Hamad MA; Seinen W; Scharnhorst V; Wulkan RW; Schönberger JP; Oeveren Wv

2012-02-01

47

Serum levels of vitamin D metabolites, calcium, phosphorus, magnesium and alkaline phosphatase in Finnish women throughout pregnancy and in cord serum at delivery.  

Science.gov (United States)

Serum concentrations of 25(OH)D, 24,25(OH)2D, 1,25(OH)2D, total calcium, protein, phosphorus, magnesium and alkaline phosphatase were measured in two groups of Finnish women throughout pregnancy and in cord serum at delivery. The autumn group delivered in August-September and the spring group in February-March. There was strong seasonal variation in the 25(OH)D concentrations in both groups. Maternal values (mean +/- s.d.) at delivery were 44.3 +/- 20.8 nmol/l in autumn and 26.0 +/- 13.0 nmol/l in spring. Fetal concentrations were 28.8 +/- 14.3 and 18.3 +/- 11.3 nmol/l, respectively. In both mothers and infants low 25(OH)D values were measured in winter. In the autumn group 7 out of 21 mothers (33 per cent) and in the spring group 17 out of 36 mothers (47 per cent) had values below 17 nmol/l, which is the lowest winter reference value recorded in our laboratory. No significant seasonal variation was observed in dihydroxylated vitamin D metabolites, although 24,25(OH)2D values were a little higher in summer than in winter. Concentrations of 1,25(OH)2D tended to rise towards delivery. Corrected calcium, magnesium and phosphorus concentrations did not change during pregnancy. Fetal calcium and phosphorus concentrations were significantly (P less than 0.001) higher than maternal ones. The data indicate that many mothers and infants have poor vitamin D status in the latitude of Finland. Our results support the concept that vitamin D supplementation should be considered in Finland for pregnant women at least in winter. PMID:3488981

Kuoppala, T; Tuimala, R; Parviainen, M; Koskinen, T; Ala-Houhala, M

1986-07-01

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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; Benito Juarez Nunes Alves de Oliveira; Duvaldo Eurides; Ednaldo Carvalho Guimarães; Luiz Antônio Franco da Silva; Lorena Borges Alves; Ana Flávia Delben Pereira Arruda; Taís Andrade Dias

2011-01-01

49

Bone-specific alkaline phosphatase protein, total alkaline phosphatase activity and lactate dehydrogenase in sera of patients with sickle cell disease.  

UK PubMed Central (United Kingdom)

Serum bone-specific alkaline phosphatase protein (bAP) was evaluated as indicator of bone turnover by immunoradiometric assay (IRMA) in twenty patients with sickle cell disease and in twenty healthy control subjects. Serum bAP was also compared with serum total alkaline phosphatase activity and serum lactate dehydrogenase in the same group. The concentrations of serum bAP and serum lactate dehydrogenase were significantly higher in the study group than in the control group (p < 0.05, p < 0.01, respectively). The serum total alkaline phosphatase activity showed no significant difference with the control healthy subjects. There was no correlation between serum bAP and total alkaline phosphatase or lactate dehydrogenase levels in the patient group. In conclusion, serum bAP protein measured by IRMA can be considered a sensitive marker of bone turnover and could be especially useful as valuable non-invasive biochemical marker for identifying sickle cell patients with skeletal complications.

Bolarin DM

2001-01-01

50

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; Isah H; Afonja O

2006-01-01

51

Effects of 60Co gamma-ray local irradiation on rat liver on alkaline phosphatase, lactate dehydrogenase and catalase in the liver and serum  

International Nuclear Information System (INIS)

[en] Rats were given a single exposure of various doses (0, 5, 50, 500, and 5000 rads) to local irradiation of 60Co ?-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

52

Osteocalcin and bone-specific alkaline phosphatase in sickle cell haemoglobinopathies.  

UK PubMed Central (United Kingdom)

Osteocalcin or bone gamma-carboxyglutamic acid (gla) protein and Bone-specific alkaline phosphatase (b-AP) total protein levels were evaluated as indicators of bone turnover in twenty patients with sickle cell haemoglobinopathies and in twenty normal healthy individuals. The serum bone-specific alkaline phosphatase total protein level was measured by immunoradiometric (IRMA) method. The concentrations of serum bone-specific alkaline phosphatase total protein were higher in the study group than in the control group [P < 0.05]. The serum osteocalcin (BGP) showed no significant difference with the control healthy subjects. There was no correlation between the serum osteocalcin and serum bone-specific alkaline phosphatase total protein in the patient group. In conclusion, serum bone-specific alkaline phosphatase total protein determined or measured by IRMA can be considered a sensitive marker of bone turnover and could be especially useful as valuable non-invasive biochemical marker for identifying sickle cell patients with bone complications.

Azinge EC; Bolarin DM

2006-06-01

53

Osteocalcin and bone-specific alkaline phosphatase in sickle cell haemoglobinopathies.  

Science.gov (United States)

Osteocalcin or bone gamma-carboxyglutamic acid (gla) protein and Bone-specific alkaline phosphatase (b-AP) total protein levels were evaluated as indicators of bone turnover in twenty patients with sickle cell haemoglobinopathies and in twenty normal healthy individuals. The serum bone-specific alkaline phosphatase total protein level was measured by immunoradiometric (IRMA) method. The concentrations of serum bone-specific alkaline phosphatase total protein were higher in the study group than in the control group [P < 0.05]. The serum osteocalcin (BGP) showed no significant difference with the control healthy subjects. There was no correlation between the serum osteocalcin and serum bone-specific alkaline phosphatase total protein in the patient group. In conclusion, serum bone-specific alkaline phosphatase total protein determined or measured by IRMA can be considered a sensitive marker of bone turnover and could be especially useful as valuable non-invasive biochemical marker for identifying sickle cell patients with bone complications. PMID:17242729

Azinge, E C; Bolarin, D M

54

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...

Min, Joo-Won; Um, Sang-Won; Yim, Jae-Jun; Yoo, Chul-Gyu; Han, Sung Koo; Shim, Young-Soo; Kim, Young Whan

55

Serum proteins, trace metals and phosphatases in psoriasis  

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Full Text Available Serum proteins, zinc, copper, acid phosphatase (AcPase) and alkaline phosphatase (AlPase) were studied in both active and remission phases of psoriasis. Data were compared with healthy controls, ?1, ? and ? globulins were high in active phase while ?1 and ? globulins were at par in remission phase. Serum copper was low but zinc and alkaline phosphatase were significantly high in both active and remission phases of the disease. Acid phosphatase level was at par in all the experimental groups. Study suggests a positive correlation of globulin, zinc and Alpase in active and remission phase of psoriasis.

Bhatnagar M; Bapna A; Khare A

1994-01-01

56

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

2012-01-01

57

Effect of cadmium and aluminum on bone alkaline and acid phosphatases. [Rats  

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Despite the large differences between Cd and Al in their physico-chemical properties, it is interesting that the two metals eventually induce osteomalacia. Therefore, the authors attempted to investigate, in vitro, the effects of Cd and Al on acid and alkaline phosphatases in rat calvarium, and in addition, to measure serum alkaline phosphatase in adult animals given Cd or Al.

Sugawara, N.; Sadamoto, T.; Sugawara, C.

1983-10-01

58

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

Science.gov (United States)

Nipah virus (NiV), Paramyxoviridae, Henipavirus, is classified as a biosafety level (BSL) 4 pathogen, along with the closely related Hendra virus (HeV). A novel serum neutralization test was developed for measuring NiV neutralizing antibodies under BSL2 conditions using a recombinant vesicular stomatitis virus (VSV) expressing secreted alkaline phosphatase (SEAP) and pseudotyped with NiV F/G proteins (VSV-NiV-SEAP). A unique characteristic of this novel assay is the ability to obtain neutralization titers by measuring SEAP activity in supernatant using a common ELISA plate reader. This confers a remarkable advantage over the first generation of NiV-pseudotypes expressing green fluorescent protein or luciferase, which require expensive and specific measuring equipment. Using panels of NiV- and HeV-specific sera from various species, the VSV-NiV-SEAP assay demonstrated neutralizing antibody status (positive/negative) consistent with that obtained by conventional live NiV test, and gave higher antibody titers than the latter. Additionally, when screening sixty-six fruit bat sera at one dilution, the VSV-NiV-SEAP assay produced identical results to the live NiV test and only required a very small amount (2?l) of sera. The results suggest that this novel VSV-NiV-SEAP assay is safe, useful for high-throughput screening of sera using an ELISA plate reader, and has high sensitivity and specificity. PMID:22115786

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

2011-11-18

59

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

UK PubMed Central (United Kingdom)

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

Kaku Y; Noguchi A; Marsh GA; Barr JA; Okutani A; Hotta K; Bazartseren B; Fukushi S; Broder CC; Yamada A; Inoue S; Wang LF

2012-01-01

60

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

 
 
 
 
61

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; Hosseinkhani H; Ismail M; Domb AJ; Omar AR; Chong PP; Hong PD; Yu DS; Farber IY

2012-01-01

62

Bone-specific alkaline phosphatase protein, total alkaline phosphatase activity and lactate dehydrogenase in sera of patients with sickle cell disease.  

Science.gov (United States)

Serum bone-specific alkaline phosphatase protein (bAP) was evaluated as indicator of bone turnover by immunoradiometric assay (IRMA) in twenty patients with sickle cell disease and in twenty healthy control subjects. Serum bAP was also compared with serum total alkaline phosphatase activity and serum lactate dehydrogenase in the same group. The concentrations of serum bAP and serum lactate dehydrogenase were significantly higher in the study group than in the control group (p < 0.05, p < 0.01, respectively). The serum total alkaline phosphatase activity showed no significant difference with the control healthy subjects. There was no correlation between serum bAP and total alkaline phosphatase or lactate dehydrogenase levels in the patient group. In conclusion, serum bAP protein measured by IRMA can be considered a sensitive marker of bone turnover and could be especially useful as valuable non-invasive biochemical marker for identifying sickle cell patients with skeletal complications. PMID:11345404

Bolarin, D M

2001-01-01

63

Phytoplankton contribution to alkaline phosphatase activity  

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A centrifugation procedure was used to partition alkaline phosphatase activity (APA) and assimilated carbon (/sup 14/C) in phytoplankton from four Michigan lakes. In all cases, although assimilated carbon partitioned nearly quantitatively, APA did not. The partitioning disparity was used to compute APA contributions by phytoplankton, nonalgal particulate matter and dissolved enzyme. Maximum possible algal contribution to total AP was consistently < 34 percent, and in some cases was as low as 5 to 6 percent. Non-algal particulate APA typically comprised 15 to 73 percent of the total APA. Filtration techniques commonly used the assess algal contibution to APA may seriously overestimate the quantity of APA directly affiliated with algal cells.

Stewart, A.J. (Oak Ridge National Lab., TN); Wetzel, R.G.

1982-02-01

64

Negative modulation of alkaline phosphatase and creatine kinase by homobrassinolide  

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Full Text Available Homobrassinolide is a plant hormone implicated in plant growth and development. Its effect on animal metabolism was less known to date. We have investigated its effect on the marker enzymes such as alkaline phosphatase and creatine kinase in selected rat tissues-brain, heart, liver, kidney, skeletal muscle and testis. Homobrassinolide was administered (66 and 330ng/ Kg body weight) intradermally in male albino wistar strain rats and changes in alkaline phosphatase and creatine kinase activities were measured. An overall reduction in both the enzyme activities occurred within 2hr of administration with few exceptions. The reaction rate constants for the enzyme activities were in the order 10-7 mM/min for alkaline phosphatase and 10-3 mM/min for creatine kinase. Time course studies indicated a decrease in enzyme activities as a function of time. Elevated hemoglobin content correlated with rise in erythrocyte number. Blood glucose level decreased by a percentage of 15.7 and 21.7 compared to control with the administration of 10?g and 50?g homobrassinolide respectively. Serum cholesterol content showed 15% decrease and 25% increase compared to control following 10?g and 50?g homobrassinolide administration. We conclude that homobrassinolide inhibited both the enzymes in the tissues and produced erythrocytosis, leukocytosis and hypoglycemia, while cellular phosphorylation status remained principally affected by this oxysterol in rat. Even though the physiological and pathological significance of these observations is not clear, it is suggested that 28-HB enriched diets may not be appropriate for higher energy related work activities.Keywords: Alkaline phosphatase; Creatine kinase; Homobrassinolide; Oxysterol; Phosphorylation; Rate constant.

G. Nirmal Kumar; S Lakshmy; K Srikumar

2011-01-01

65

[Alkaline phosphatase in the blood and lymph in diffuse peritonitis  

UK PubMed Central (United Kingdom)

Activity of alkaline phosphatase was studied in blood and lymph of 52 dogs with experimental peritonitis and in 26 patients with the drained thoracic duct and in 26 patients without lymphodrainage. Pronounced enzyme intoxication was revealed. Under physiological conditions alkaline phosphatase is transported by both blood and lymphatic bed. In peritonitis the lymphatic route of the enzyme transport gets considerably greater. Drainage of the thoracic duct gives good detoxicating effect (decreased alkaline phosphatase activity).

Nikolaeva EV

1985-10-01

66

Modifications induced by ascorbic acid on alkaline phosphatase fluorescence.  

UK PubMed Central (United Kingdom)

Ascorbic acid, isoascorbic acid and dehydroascorbic acid quench the tryptophyl fluorescence of alkaline phosphatase. The quenching is protein aspecific, although its extent reflects the different inhibitory efficiency of the compounds. The kinetic inactivation and emission deactivation of alkaline phosphatase isoenzymes present also striking similarities. The fluorescence modifications, moreover, show a particular pattern, indicative of a transition phenomenon. The quenching effects displayed by the ascorbic system on alkaline phosphatase can then supply an interesting insight into other aspects of the inhibitor-enzyme interaction.

Martorana GE; Meucci E; Miggiano GA; Mordente A; Ursitti A; Castelli A

1984-09-01

67

Alkaline phosphatases in tissues and sera of cats.  

UK PubMed Central (United Kingdom)

The numbers and widths of bands of alkaline phosphatase (ALP) activity in polyacrylamide gels and the comparison of their electrophoretic mobility to that of a reference substance (Rf value) were found to be reliable aids in the identificaiton of various isoenzymes in in serum and organ extracts from cats. The hepatic isoenzyme was identified in sera of clinically normal adult cats, pregnant cats late in gestation, and cats with common bile duct occlusion. In addition to the hepatic isoenzyme, placental ALP was found late in gestation in sera from queens. Sera from kittens less than 15 weeks of age contained only the osseous ALP isoenzyme.

Everett RM; Duncan JR; Prasse KW

1977-10-01

68

Human placental alkaline phosphatase in liver and intestine  

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Three distinct forms of human alkaline phosphatase, presumably isozymes, are known, each apparently associated with a specific tissue. These are placental, intestinal, and liver (kidney and bone). The authors have used a specific immunoassay and HPLC to show that placental alkaline phosphatase is also present in extracts of liver and intestine in appreciable amounts.

Garattini, E.; Margolis, J.; Heimer, E.; Felix, A.; Udenfriend, S.

1985-09-01

69

Intestinal calcium and phosphate transport and intestinal alkaline phosphatase.  

UK PubMed Central (United Kingdom)

The aim of the study was to investigate the link, if any, between alkaline phosphatase activity and intestinal calcium and phosphate transport using the hypophysectomized (HX) rat model. Ionic transport was evaluated by the in situ ligated loop technique. Hypophysectomy (HX) resulted in a decrease in both duodenal and jejunal alkaline phosphatase activity but did not alter the active transport of calcium and phosphate. Vitamin D depletion (-D), suppressed intestinal transport in the HX rat without altering alkaline phosphatase activity. Repletion of 1,25(OH)2D3 in the -DHX rat resulted in an increase in active transport of calcium and phosphate without altering the alkaline phosphatase activity. Thus, using the HX animal model, we were able to differentiate intestinal alkaline phosphatase activity from the overall intestinal transport of calcium and phosphate.

Yeh JK; Aloia JF

1986-09-01

70

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

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

Jemmerson, R.; Low, M.G.

1987-09-08

71

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

International Nuclear Information System (INIS)

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

1987-09-08

72

Calcium pyrophosphate dihydrate (CPPD) crystal dissolution by alkaline phosphatase: interaction of alkaline phosphatase on CPPD crystals.  

UK PubMed Central (United Kingdom)

OBJECTIVE: As alkaline phosphatase (ALP) can dissolve calcium pyrophosphate dihydrate (CPPD) crystals, and as dissolution is facilitated when the enzyme is proximate to the crystals, we studied the mechanism of ALP interaction with CPPD crystals in vitro. METHODS: ALP was incubated with CPPD crystals in an in vitro model system. Fluorescein isothiocyanate conjugated alkaline phosphatase (FITC-ALP), alkaline phosphatase product staining of calcium pyrophosphate dihydrate (CPPD) crystals and scanning electron microscopy were used to visualize ALP-CPPD crystal interactions. RESULTS: ALP preferentially binds to the small end faces (optical 010 faces) of CPPD crystals. Etch pits indicative of dissolution were demonstrated coexistent with ALP crystal binding and ALP pyrophosphohydrolytic activity. CONCLUSION: ALP binding to CPPD crystals is preferential for the smallest end faces (optical 010 faces). As ALP crystal binding is altered by ions but not by heat inactivation of ALP, ALP-CPPD crystal binding is considered a nonenzymatic mechanism distinct from ALP pyrophosphohydrolytic activity. Our study demonstrates that ALP binds and dissolves CPPD crystals in a stereoselective manner. This suggests that the CPPD crystal dissolution rate is limited by the availability of surface area on the crystal faces most susceptible to ALP binding.

Shinozaki T; Xu Y; Cruz TF; Pritzker KP

1995-01-01

73

A study of the alkaline and acid phosphatase activities in acute uranium intoxication  

International Nuclear Information System (INIS)

Comparative study of the ability of the sodium salt of diethylbarbituric acid and acetazolamide to protect the kidneys is conducted under conditions of acute uranium intoxication in rats. The parameters studied are alkaline and acid phosphatase activities in the serum and urine and phosphatase activity in the kidneys (histochemically as described by Gomori) followed up until the 30th day after the total uranyl acetate dose was reached (2 or 7 mg per kg bodyweight). Either compound exerted only minor effect on serum alkaline phosphatase activity. Sodium diethylbarbiturate induced distinct fluctuations in urinary alkaline phosphatase activity throughout the entire study period, but the differences never reached statistical significance. Acetazolamide caused essential decrease in urinary alkaline phosphatase activity. In either case renal tissue protection from the action of the uranyl ion may be suggested. This assumption is supported by the histochemical analysis. The compounds appeared to have no effect on serum acid phosphatase activity which showed high variability both in control and in treated rats. (Ch.K.)

1975-01-01

74

Alkaline phosphatase dissolves calcium pyrophosphate dihydrate crystals.  

UK PubMed Central (United Kingdom)

We have shown that yeast pyrophosphatase dissolves calcium pyrophosphate dihydrate (CPPD) crystals in solutions. In this investigation we demonstrate that alkaline phosphatase (ALP) effectively dissolves CPPD crystals in vitro. CPPD dissolution by ALP had a pH optimum of 7.4, which is the optimum pH for its pyrophosphatase (PPiase) activity. The CPPD dissolution and PPiase activity by ALP are magnesium dependent, whereas its phosphoester hydrolytic activity is not. Calcium, which inhibited the enzymatic CPPD dissolution and PPiase activity of ALP had no effect on its phosphoester hydrolytic activity. These data indicate that PPiase activity of ALP is responsible for CPPD dissolution and not its phosphoester hydrolytic activity. Matrix molecules such as proteoglycans and chondroitin sulfate had no effect on the enzymatic and nonenzymatic dissolution of CPPD crystals. ALP acted more effectively on CPPD crystals than on soluble pyrophosphate relative to yeast PPiase. Our data suggest that chondrocyte ALP may play an important role in the dissolution of CPPD crystals in cartilage.

Xu Y; Cruz TF; Pritzker KP

1991-10-01

75

Alkaline phosphatase as a periodontal disease marker  

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Full Text Available 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 measures of the inflammatory activity occurring in the adjacent periodontal tissues. Objective: The aim of this study was to assess the total activity of ALP in the GCF collected from healthy sites, sites with gingivitis and with chronic adult periodontitis. An attempt was also made to establish the correlation of ALP activity with plaque index, gingival index, bleeding index and probing depth. Materials and Methods: A total of 18 patients were divided into three groups: viz., healthy sites, Group I; gingivitis, Group II; chronic periodontitis, Group III. Clinical parameters like plaque index, bleeding index, gingival index and probing depth were recorded. The ALP level in GCF of all three groups was determined by spectrophotometric analysis. Results: Total enzyme activity of ALP was significantly higher in periodontitis as compared with that in healthy and gingivitis sites, and was significantly and positively correlated with probing depth. Conclusion: ALP can be considered as a periodontal disease marker as it can distinguish between healthy and inflamed sites. However, to better define its capacity for periodontal diagnosis, additional longitudinal studies are required.

Malhotra Ranjan; Grover Vishakha; Kapoor Anoop; Kapur Rupika

2010-01-01

76

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

International Nuclear Information System (INIS)

[en] 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

77

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

Energy Technology Data Exchange (ETDEWEB)

A cDNA clone for human adult intestinal alkaline phosphatase (ALP) (orthophosphoric-monoester phosphohydrolase (alkaline optimum); EC 3.1.3.1) was isolated from a lambdagt11 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.

Henthorn, P.S.; Raducha, M.; Edwards, Y.H.; Weiss, M.J.; Slaughter, C.; Lafferty, M.A.; Harris, H.

1987-03-01

78

An evaluation of the effect of age and the peri-parturient period on bone metabolism in dairy cows as measured by serum bone-specific alkaline phosphatase activity and urinary deoxypyridinoline concentration.  

UK PubMed Central (United Kingdom)

Various biochemical markers help to evaluate the state of bone turnover in humans and could be used during the peri-parturient period in dairy cows when calcium (Ca) metabolism changes dramatically. To investigate this, the peri-partum characteristics of serum bone-specific alkaline phosphatase (BAP) and urinary deoxypyridinoline (DPD) were investigated. Both serum BAP activity and urinary DPD concentrations were increased and demonstrated wide variability in younger animals, and these findings were consistent with other bone turnover markers. Around the time of parturition, serum Ca concentration and serum BAP activity in multiparous cows were significantly lower than in primiparous cows, but urinary DPD concentration was unchanged. The use of BAP as a bone formation marker appears to be valuable for evaluating bone remodelling status in cows, but the specificity of the test needs to be confirmed. The DPD/BAP ratio around parturition demonstrated a clear difference in bone turnover status between the two parity groups with multiparous cows demonstrating increased signs of bone resorption compared with primiparous cows, corresponding to the Ca requirement for milk production. In future studies, the applicability of the ratio of bone resorption marker to bone formation marker should be evaluated for bone turnover assessment.

Sato R; Onda K; Kato H; Ochiai H; Kawai K; Iriki T; Kaneko K; Yamazaki Y; Wada Y

2013-08-01

79

An evaluation of the effect of age and the peri-parturient period on bone metabolism in dairy cows as measured by serum bone-specific alkaline phosphatase activity and urinary deoxypyridinoline concentration.  

Science.gov (United States)

Various biochemical markers help to evaluate the state of bone turnover in humans and could be used during the peri-parturient period in dairy cows when calcium (Ca) metabolism changes dramatically. To investigate this, the peri-partum characteristics of serum bone-specific alkaline phosphatase (BAP) and urinary deoxypyridinoline (DPD) were investigated. Both serum BAP activity and urinary DPD concentrations were increased and demonstrated wide variability in younger animals, and these findings were consistent with other bone turnover markers. Around the time of parturition, serum Ca concentration and serum BAP activity in multiparous cows were significantly lower than in primiparous cows, but urinary DPD concentration was unchanged. The use of BAP as a bone formation marker appears to be valuable for evaluating bone remodelling status in cows, but the specificity of the test needs to be confirmed. The DPD/BAP ratio around parturition demonstrated a clear difference in bone turnover status between the two parity groups with multiparous cows demonstrating increased signs of bone resorption compared with primiparous cows, corresponding to the Ca requirement for milk production. In future studies, the applicability of the ratio of bone resorption marker to bone formation marker should be evaluated for bone turnover assessment. PMID:23422881

Sato, Reiichiro; Onda, Ken; Kato, Hajime; Ochiai, Hideharu; Kawai, Kazuhiro; Iriki, Tsunenori; Kaneko, Kazuyuki; Yamazaki, Yukio; Wada, Yasunori

2013-02-17

80

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; Amir Mahdi Imani; Sasan Najjari Asl; Hossein H.Farhang; Khazar Ghasempour; Ezzatzadeh; Nava Ainechi

2013-01-01

 
 
 
 
81

Altering of the metal specificity of Escherichia coli alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Analysis of sequence alignments of alkaline phosphatases revealed a correlation between metal specificity and certain amino acid side chains in the active site that are metal-binding ligands. The Zn(2+)-requiring Escherichia coli alkaline phosphatase has an Asp at position 153 and a Lys at position 328. Co(2+)-requiring alkaline phosphatases from Thermotoga maritima and Bacillus subtilis have a His and a Trp at these positions, respectively. The mutations D153H, K328W, and D153H/K328W were induced in E. coli alkaline phosphatase to determine whether these residues dictate the metal dependence of the enzyme. The wild-type and D153H enzymes showed very little activity in the presence of Co(2+), but the K328W and especially the D153H/K328W enzymes effectively use Co(2+) for catalysis. Isothermal titration calorimetry experiments showed that in all cases except for the D153H/K328W enzyme, a possible conformation change occurs upon binding Co(2+). These data together indicate that the active site of the D153H/K328W enzyme has been altered significantly enough to allow the enzyme to utilize Co(2+) for catalysis. These studies suggest that the active site residues His and Trp at the E. coli enzyme positions 153 and 328, respectively, at least partially dictate the metal specificity of alkaline phosphatase.

Wojciechowski CL; Kantrowitz ER

2002-12-01

82

Effect of dietary sorbitol on alkaline phosphatase and glucose-6-phosphatase in the mouse.  

UK PubMed Central (United Kingdom)

Mice fed a sorbitol-enriched diet show measurable increases of alkaline phosphatase, glucose-6-phosphatase, and glucose-6-phosphate in the liver. The increases are positively correlated with duration of the sorbitol feeding program. The results could imply that sorbitol promotes glycogen synthesis.

Zanobini A; Firenzuoli AM

1982-01-01

83

Radiation effects on alkaline phosphatase and glucose-6-phosphatase in anatomically different regions of mouse intestine  

International Nuclear Information System (INIS)

[en] The effect of gamma irradiation on alkaline phosphatase and glucose-6-phosphate has been studied in three anatomically different regions of the small intestine at a surface dose of 400 R. Both the enzymatic activities were shown to be enhanced in duodenum, jejunum and ileum 24 hours after irradiation. The activity of alkaline phosphatase on day 3 tended to be low as compared to day 1 post irradiation, but glucose-6-phosphatase continued to rise even after day 3. Maximum rise of glucose-6-phosphatase was observed in the jejunum. On day 9, alkaline phosphatase was diminished below the controls in the whole of intestine, but appeared to be normal on day 10. Glucose-6-phosphatase in duodenum and jejunum on the other hand was comparable to that of control mice; but in ileum, the activity of this enzyme was below the normal values. Physiological significance of these enzymes in intestine has been discussed. (orig.)[de] Die Wirkung von Gammastrahlung auf alkalische Phosphatase und Glucose-6-phosphatase wurde bei einer Oberflaechendosis von 400 R an drei anatomisch verschiedenen Teilen des Duenndarms untersucht. Eine Steigerung der Aktivitaet beider Enzyme in Duodenum, Jejunum und Ileum konnte 24 Stunden nach Bestrahlung festgestellt werden. Am dritten Tag war die Aktivitaet der alkalischen Phosphatase eher niedrig, verglichen mit dem ersten Tag nach der Bestrahlung; aber die Aktivitaet der Glucose-6-phosphatase stieg weiterhin an, selbst nach dem dritten Tag. Ihre maximale Zunahme wurde im Jejunum beobachtet. Am neunten Tag sank die alkalische Phosphatase im gesamten Darm unter den Kontrollwert, doch schien sie am zehnten Tag normal zu sein. Dagegen hatte die Glucose-6-phosphatase im Duodenum und Jejunum mit den Kontrollmaeusen vergleichbare Werte; im Ileum aber war die Aktivitaet dieses Enzyms unterhalb der Normalwerte. Die physiologische Bedeutung der beiden Enzyme im Darm wird eroertert. (orig.)

1975-01-01

84

Regulation of alkaline phosphatase: implications for calcium pyrophosphate dihydrate crystal dissolution and other alkaline phosphatase functions.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Alkaline phosphatase (ALP), an enzyme with pyrophosphatase (PPiase) activity can dissolve calcium pyrophosphate dihydrate (CPPD) crystals. We studied the effects of enzyme inhibitors such as bisphosphonates, orthovanadate, calcium, cadmium, and ascorbic acid on PPiase activity of ALP as well as on phosphate ester hydrolysis (Pase) activity and compared these effects to those on CPPD crystal dissolution. METHOD: An in vitro model system for crystal enzyme interaction was used to assess CPPD crystal dissolution. RESULTS: Bisphosphonates inhibited ALP Pase activity more than ALP PPiase activity at the same concentrations. Calcium inhibited ALP PPiase activity, but not ALP Pase activity. Orthovanadate and cadmium inhibited ALP PPiase activity more than ALP Pase at the same concentrations. The inhibition rates of ALP PPiase at the same concentrations were orthovanadate > cadmium > calcium. Although ALP Pase activity was not inhibited, at high concentrations, ascorbic acid slightly inhibited ALP PPiase activity. Bisphosphonates at high concentrations inhibited ALP CPPD crystal dissolution. The strong inhibitory effects of bisphosphonates on ALP CPPD crystal dissolution compared to those on ALP PPiase activity suggest that bisphosphonates inhibit crystal dissolution by their affinity for the CPPD crystal surface. Calcium, orthovanadate, and cadmium inhibited ALP CPPD dissolution. The inhibition rates of ALP CPPD dissolution at the same concentrations were cadmium > calcium > orthovanadate. Ascorbic acid at high concentrations enhanced ALP CPPD dissolution. CONCLUSION: These effects of different inhibitors on ALP PPiase and CPPD dissolution suggest that ALP CPPD crystal dissolution depends on binding of ALP CPPD crystals as well as the PPiase activity of the bound ALP. Because of its ubiquitous and broad phosphatase activity including PPiase activity, ALP may have a critical role in cell energy metabolism.

Shinozaki T; Pritzker KP

1996-04-01

85

Plasma intestinal alkaline phosphatase isoenzymes in neonates with bowel necrosis.  

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AIM--To determine if the intestinal isoenzymes of alkaline phosphatase (ALP) are biochemical markers of bowel necrosis in neonates. METHODS--Plasma ALP isoenzymes were measured in 22 babies with bowel necrosis, histologically confirmed, and in 22 matched controls. The isoenzymes were also measured i...

McLachlan, R; Coakley, J; Murton, L; Campbell, N

86

Clinical significance of an ultrafast alkaline phosphatase isoenzyme.  

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We have studied five patients who have exhibited an unusual alkaline phosphatase isoenzyme (ALP EC 3.1.3.1) migrating in an ultrafast position electrophoretically on cellulose acetate. This ALP isoenzyme has been identified in patients with benign and malignant liver diseases. In addition, a number ...

Koett, J; Howell, J; Wolf, P L

87

Bone alkaline phosphatase and mortality in dialysis patients.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVES: Serum alkaline phosphatase (AP) is associated with vascular calcification and mortality in hemodialysis patients, but AP derives from various tissues of origin. The aim of this study was to assess the effect of bone-specific AP (BAP) on morbidity and mortality in dialysis patients. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: From a prospective cohort study of incident dialysis patients in The Netherlands, all patients with measured BAP at 12 months after the start of dialysis (baseline) were included in the analysis (n = 800; mean age, 59 ± 15 years; mean BAP = 18 ± 13 U/L). By Cox regression analyses, we assessed the impact of BAP levels on short-term mortality (6 months) and longer-term mortality (4-year follow-up). RESULTS: High levels of BAP strongly affected short-term mortality. After adjustment for confounders, patients in the highest BAP tertile had a 5.7-fold increased risk of death within 6 months compared with patients in the lowest tertile. The effect applied to both cardiovascular and noncardiovascular mortality. Furthermore, high levels of BAP were associated with increased cardiovascular mortality in the longer term. In comparison with total AP, the effect sizes related to clinical outcomes were much higher for BAP. CONCLUSIONS: High levels of BAP were strongly associated with short-term mortality in dialysis patients, pointing out the important impact of bone turnover. Longitudinal assessments of BAP may be useful for the treatment monitoring in clinical practice in dialysis patients.

Drechsler C; Verduijn M; Pilz S; Krediet RT; Dekker FW; Wanner C; Ketteler M; Boeschoten EW; Brandenburg V

2011-07-01

88

SERUM PROTEINS, TRANSAMINASES AND PHOSPHATASES IN MALNUTRITION  

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Full Text Available The levels of serum tota1 protein, albumin, transaminases and phosphatases were estimated in a group of children with severe Marasmus or mild malnutrition in order to identify some of the associated deficiencies in these syndromes. The biochemical pattern was similar in the normal and malnourished children.

H. Mohammadiha; J. Nahani; M. Rafii; N. Mohagheghpour.

1976-01-01

89

Secreted phosphoprotein-24?kDa (Spp24) attenuates BMP-2-stimulated Smad 1/5 phosphorylation and alkaline phosphatase induction and was purified in a protective complex with alpha2 -Macroglobulins From Serum.  

UK PubMed Central (United Kingdom)

Secreted phosphoprotein-24?kDa (Spp24) binds cytokines of the bone morphogenetic protein/transforming growth factor-? (BMP/TGF?) superfamily and is one of the most abundant serum phosphoproteins synthesized by the liver. Little is known about how Spp24 binding affects BMP signal transduction and osteoblastic differentiation or how this labile protein is transported from the liver to remote tissues, such as bone. When Spp24 was administered to W-20-17 mesenchymal stem cells with rhBMP-2, short-term Smad1/5 phosphorylation was inhibited, intermediate-term alkaline phosphatase (ALP) induction was blunted, and long-term mineralization was unaffected. This supports the hypothesis that Spp24 proteolysis restricts the duration of its regulatory effects, but offers no insight into how Spp24 is transported intact from the liver to bone. When Spp24 was immunopurified from serum and subjected to native PAGE and Western blotting, a high molecular weight band of >500?kDa was found. Under reducing SDS-PAGE, a 24?kDa band corresponding to monomeric Spp24 was liberated, suggesting that Spp24 is bound to a complex linked by disulfide bonds. However, such a complex cannot be disrupted by 60?mM EDTA under non-reducing condition or in purification buffers containing 600?mM NaCl and 0.1% Tween-20 at pH 2.7-8.5. LC-MS/MS analysis of affinity-purified, non-reducing SDS-PAGE separated, and trypsin digested bands showed that the Spp24 was present in a complex with three ?(2) -macroglobulins (?(2) -macroglobulin [?(2) M], pregnancy zone protein [PZP] and complement C3 [C3]), as well as ceruloplasmin and the protease inhibitor anti-thrombin III (Serpin C1), which may protect Spp24 from proteolysis.

Zhao KW; Murray SS; Murray EJ

2013-02-01

90

A simple radioimmunoassay of human placental alkaline phosphatase (Regan isoenzyme) using specific antibody polymers.  

UK PubMed Central (United Kingdom)

Rabbit antiserum against highly purified high-molecular-weight B-variant of human placental alkaline phosphatase (M.W. 200,000) was rendered monospecific by absorption with polymerized pooled male serum proteins; the absorbed antiserum was then polymerized with ethyl chloroformate and used in radioimmunoassay as a stable solid-phase immunoabsorbent. Homogeneous preparation of the enzyme, with a specific activity of 477 mumoles phenol per mg per min, was also obtained by absorbing the chromatographically purified enzyme with polymerized rabbit antiserum directed to whole human serum proteins; the pure enzyme was then labeled with 125-I as the tracer retaining at least 80% of its antigenicity. Only a minute quantity of the polymerized antibody particles is required for each assay in admixture with the labeled and unlabeled enzyme. By adding a small amount of starch-gel particles before low-speed centrifugation, complete phase separation was achieved. The radioimmunoassay could detect 0.4 to 0.8 ng enzyme protein per tube, which is comparable to the sensitivity achieved by enzymic assays. However, radioimmunoassay is advantageous over the enzymic assay in being direct, specific (no interference by the nonplacental-type alkaline posphatase), and capable of detecting both catalytically active and inactive forms of the enzyme. Native variants of placental-type alkaline phosphatase including Regan isoenzyme and Nagao isoenzyme (D-phenotype of normal placental alkaline phosphatase), could thus be directly determined by this procedure in the clinical specimens.

Chang CH; Raam S; Angellis D; Doellgast G; Fishman WH

1975-07-01

91

Placental alkaline phosphatase in patients with seminoma measured by ELISA as compared to RIA.  

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Full Text Available Placental alkaline phosphatase (PLAP) has been shown to be a reliable tumor marker in the management of patients with seminoma. Radioimmunoassay (RIA) and Enzyme linked immunoassay (ELISA) procedures were compared for PLAP levels in serum of seminoma patients. The statistical analysis showed excellent correlationship between the two. It is evident from the results that performance of ELISA in terms of sensitivity specificity, precision, accuracy and reproducibility is equivalent to or better than RIA.

Redkar S; Damle S

1997-01-01

92

Placental alkaline phosphatase in patients with seminoma measured by ELISA as compared to RIA.  

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Placental alkaline phosphatase (PLAP) has been shown to be a reliable tumor marker in the management of patients with seminoma. Radioimmunoassay (RIA) and Enzyme linked immunoassay (ELISA) procedures were compared for PLAP levels in serum of seminoma patients. The statistical analysis showed excellent correlationship between the two. It is evident from the results that performance of ELISA in terms of sensitivity specificity, precision, accuracy and reproducibility is equivalent to or better than RIA. PMID:9491665

Redkar, S L; Damle, S R

1997-06-01

93

The relationship between glucose metabolism, metabolic syndrome, and bone-specific alkaline phosphatase: a structural equation modeling approach.  

Science.gov (United States)

Context: Serum alkaline phosphatase plays a role in vascular calcification. It is found in various tissues, whereas bone-specific alkaline phosphatase (BAP) more specifically reflects mineral metabolism. The relationship of serum alkaline phosphatase (total and bone-specific) with diabetes and metabolic syndrome (MetS), 2 major risk factors of vascular calcification, is largely unknown. Objective: We aimed to investigate the relationships between glucose metabolism, components of the MetS, and alkaline phosphatase. Design and Setting: This was a cross-sectional study of a nationally representative sample of the U.S. population in 1999 through 2004. Participants: Participants were 3773 nondiabetic participants of the National Health and Nutrition Examination Survey 1999-2004. Main Outcome Measures: We measured serum BAP and total alkaline phosphatase. Results: In multivariable linear regression, updated homeostasis model assessment (HOMA2) for insulin resistance (? = 0.068), HOMA2 for ?-cell function (? = 0.081), insulin (? = 0.065), mean arterial pressure (? = 0.15), and high-density lipoprotein (HDL)-cholesterol (? = 0.209) were positively associated with BAP, whereas HOMA2 for insulin sensitivity (? = -0.065) was negatively associated with BAP. On the other hand, only mean arterial pressure and HDL-cholesterol were significantly associated with total alkaline phosphatase. Moreover, a structural equation model revealed that hypertension, low HDL, and insulin resistance had significant direct effects on serum BAP levels, whereas obesity and inflammation might have indirect effects on serum BAP levels. The overall model showed very good fit to the data (comparative fit index = 0.995, root mean square error of approximation = 0.037, and standardized root mean square residual = 0.006). Conclusion: Glucose metabolism and MetS are significantly related to serum BAP levels. How BAP mediates vascular calcification in diabetes and MetS warrants further studies. PMID:23796564

Cheung, Ching-Lung; Tan, Kathryn C B; Lam, Karen S L; Cheung, Bernard M Y

2013-06-24

94

Characterization of alkaline phosphatase inactivation by ascorbic acid.  

UK PubMed Central (United Kingdom)

Ascorbic acid, isoascorbic acid and dehydroascorbic acid inhibit bovine kidney alkaline phosphatase activity. Ascorbic acid free radicals seem not to be involved. Dialysis does not make the inactivation reversible. A competitive mechanism can be inferred from experiments with phosphate and substrates, which block the activity decay. The influence of temperature, pH, other inhibitors and tertiary structure modifications on the inactivation process is also investigated.

Miggiano GA; Mordente A; Martorana GE; Meucci E; Castelli A

1984-09-01

95

Effect of magnesium on the properties of zinc alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase of Escherichia coli, isolated by procedures which do not alter its intrinsic metal content, contains 4.0 +/- 0.3 g-atoms of tightly bound zinc per mole (Kd less than 1 muM) and 1.3 +/- 0.2 g-atoms of magnesium per mole (Bosron, W.F., Kennedy, F.S., and Vallee, B.L. (1975), Biochemistry 14, 2275-2282). Importantly, the binding of magnesium is dependent both upon pH and zinc content. Hence, the failure to assign the maximal magnesium stoichiometry to enzyme isolated by conventional procedures may be considered a consequence of the conditions chosen for optimal bacterial growth and purification of the enzyme which are not the conditions for optimal binding of magnesium to alkaline phosphatase. Under the conditions employed for the present experimental studies, a maximum of six metal sites are available to bind zinc and magnesium, i.e., four for zinc and two for magnesium. Magnesium alone does not activate the apoenzyme, but it regulates the nature of the zinc-dependent restoration of catalytic activity to apophosphatase, increasing the activity of enzyme containing 2-g-atoms of zinc five-fold and that of enzyme containing 4-g-atoms of zinc 1.4-fold. Moreover, hydrogen-tritium exchange reveals the stabilizing effects of magnesium on the structural properties of phosphatase. However, neither the KM for substrate nor the phosphate binding stoichiometry and Ki are significantly altered by magnesium. Hence, magnesium, which is specificially bound to the enzyme, both stabilizes the dynamic protein structure and regulates the expression of catalytic activity by zinc in alkaline phosphatase.

Bosron WF; Anderson RA; Falk MC; Kennedy FS; Vallee BL

1977-02-01

96

As fosfatases alcalinas, transaminases e gama-glutamil-transferase séricas em pacientes epilépticos tratados com carbamazepina The serum alkaline phosphatases, transaminases, and gamma-glutamil transferases in epileptic patients treated with carbamazepine  

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Full Text Available INTRODUÇÃO: A carbamazepina é a droga utilizada no tratamento de pacientes com epilepsia parcial (ou focal) secundariamente generalizada. Apesar do uso terapêutico, este fármaco tem sido implicado no aumento das atividades séricas de algumas enzimas. Alguns autores descreveram valores de prevalência de 7,7%, 13% e 22% para aumento de atividade das fosfatases alcalinas séricas (FA ou EC 3.1.3.1). A divergência de resultados também foi encontrada para as atividades da gama-glutamil-transferase sérica (gama-glutamil transferase ou GGT ou EC 2.3.2.2). OBJETIVO: Assim, a meta desta pesquisa é determinar, dentre outros objetivos, a freqüência de alterações nas atividades das FA, GGT e transaminases (AST, aspartato-amino-transferase, EC 2.6.1.1; e ALT, alanina-amino-transferase, EC 2.6.1.2) de uma amostra de pacientes do ambulatório de epilepsia em Salvador, Bahia. MATERIAL E MÉTODOS: O desenho do estudo é descritivo do tipo série de casos, aprovado pelo Comitê de Ética local, no qual uma amostra de conveniência de 52 pacientes epilépticos de acompanhamento ambulatorial foi obtida sem interferência dos pesquisadores. Estes pacientes foram organizados por faixa etária de 12 a 30 e de 31 a 90 anos e, subdivididos por tempo de monoterapia com carbamazepina. As atividades séricas das enzimas GGT, FA, AST e ALT foram determinadas. RESULTADOS: As proporções de alterações por variáveis foram descritas: 42% para as FA, 18% para as GGT, 2% para as ALT e 12% para as AST, respectivamente. A faixa etária de 12 a 30 anos apresentou 56% de alterações nas FA enquanto que aquela de 31 a 90 anos, apenas 18%. CONCLUSÃO: Nós concluímos que as enzimas FA, GGT, AST e ALT apresentaram maiores freqüências de alterações de suas atividades naqueles pacientes com idade igual ou inferior a 30 anos, sendo que as FA apresentaram maiores valores.INTRODUCTION: Carbamazepine is the drug of choice used in the treatment of patients with partial (or focal) epilepsy with secondary generalization. Despite its therapeutical use, this drug has been implicated in the increase of serum activities in some enzymes. Some authors have described prevalence values of 7.7%, 13%, and 22% for the increase of activity of serum alkaline phosphatases (AF or EC 3.1.3.1). A divergence in the results was also found for the activities of the serum g-glutamil transferase (gamma-glutamil transferase or GGT, or EC 2.3.2.2). OBJECTIVE: Hence, among other objectives, the aim of this research is to determine the frequency of sample alterations in serum enzymatic activities of AF, GGT and transaminases (AST, aspartate amino-transferase, EC 2.6.1.1; and ALT, alanine-amino-transferase, EC 2.6.1.2) in epilepsy ward patients in Salvador, Bahia. MATERIAL AND METHODS: The design of the study is descriptive and it is a case series type. It has been approved by the local Ethics Committee. In this study, a convenience sample of 52 epileptic patients who receive ambulatory care was obtained without interference by the researchers. These patients were divided according to age groups of 12 to 30 years and 31 to 90 years, which were then subdivided according to the period of monotherapy with carbamazepine. The serum activities of the enzymes GGT, AF, AST and ALT were determined. RESULTS: The ratios of alterations per variables were described: 42% for the FA, 18% for the GGT, 2% for the ALT, and 12% for the AST respectively. The age group of 12 to 30 years presented 56% of alterations in the AF while the group of 31 to 90 years presented only 18% of alterations. CONCLUSION: We conclude that the enzymes AF, GGT, AST, and ALT presented higher frequencies of alterations of their activities in those patients with age equal to or below 30 years, while the AF presented higher values.

Helder Jacobina Santos; Antonio de Souza Andrade Filho; Olivia Lordelo Sanches; Tiago Spolador; Luís Erlon Araújo Rodrigues

2006-01-01

97

A specific alkaline phosphatase from Saccharomyces cerevisiae with protein phosphatase activity.  

Science.gov (United States)

In this paper, specific PHO13 alkaline phosphatase from Saccharomyces cerevisiae was demonstrated to possess phosphoprotein phosphatase activity on the phosphoseryl proteins histone II-A and casein. The enzyme is a monomeric protein with molecular mass of 60 kDa and hydrolyzes p-nitrophenyl phosphate with maximal activity at pH 8.2 with strong dependence on Mg2+ ions and an apparent Km of 3.6 x 10(-5) M. No other substrates tested except phosphorylated histone II-A and casein were hydrolyzed at any significant rate. These data suggest that the physiological role of the p-nitrophenyl phosphate-specific phosphatase may involve participation in reversible protein phosphorylation. 1988 Federation of European Microbiological Societies. PMID:9561742

Tuleva, B; Vasileva-Tonkova, E; Galabova, D

1998-04-01

98

A specific alkaline phosphatase from Saccharomyces cerevisiae with protein phosphatase activity.  

UK PubMed Central (United Kingdom)

In this paper, specific PHO13 alkaline phosphatase from Saccharomyces cerevisiae was demonstrated to possess phosphoprotein phosphatase activity on the phosphoseryl proteins histone II-A and casein. The enzyme is a monomeric protein with molecular mass of 60 kDa and hydrolyzes p-nitrophenyl phosphate with maximal activity at pH 8.2 with strong dependence on Mg2+ ions and an apparent Km of 3.6 x 10(-5) M. No other substrates tested except phosphorylated histone II-A and casein were hydrolyzed at any significant rate. These data suggest that the physiological role of the p-nitrophenyl phosphate-specific phosphatase may involve participation in reversible protein phosphorylation. 1988 Federation of European Microbiological Societies.

Tuleva B; Vasileva-Tonkova E; Galabova D

1998-04-01

99

Activation of Pyrococcus furiosus alkaline phosphatase by divalent metal ions.  

UK PubMed Central (United Kingdom)

Treatment of a hyperthermophilic enzyme, alkaline phosphatase from Pyrococcus furiosus (PfuAP), with EDTA completely deactivated PfuAP, indicating that the presence of one or more divalent metal ions is essential for its catalytic activity. Subsequent addition of various divalent metal ions to the apoprotein recovered the enzymatic activity and, in particular, the addition of Co(II) resulted in an over 50-fold increase in activity compared with PfuAP before EDTA treatment. Intriguingly, PfuAP with Co(II) exhibited weaker stability toward heat treatment, suggesting that Co(2+) destabilizes the tertiary structure of PfuAP at high temperature.

Minamihata K; Goto M; Kamiya N

2012-11-01

100

[Immunochemical properties of the extracellular hydrolases (protease and alkaline phosphatase) of Coccidioides immitis  

UK PubMed Central (United Kingdom)

Extracellular hydrolases (protease and alkaline phosphatase) of the coccidioidal fungus possessed antigenic properties and caused production of the corresponding antibodies. Phosphatase-antiphosphatase-substrate system apparently has future prospects for the elaboration of immunobiochemical methods for the diagnosis of coccidioidomycosis.

Klimova IM; Rogozhkina NM

1977-08-01

 
 
 
 
101

Alkaline phosphatase grafting on bioactive glasses and glass ceramics.  

UK PubMed Central (United Kingdom)

Bone integration of orthopaedic or dental implants and regeneration of damaged bone at the surgical site are still unresolved problems in prosthetic surgery. For this reason, biomimetic surfaces (i.e. both inorganic and biological bioactive surfaces) represent a challenge for bone implantation. In this research work a hydrolase enzyme (alkaline phosphatase) was covalently grafted to inorganic bioactive glass and glass ceramic surfaces, in order to impart biological bioactivity. The functionalized samples were analysed by means of X-ray photoelectron spectroscopy in order to verify enzyme presence on the surface. Enzyme activity was measured by means of UV-visual spectroscopy after reaction with the natural substrate. Scanning electron microscopy-energy-dispersive spectroscopy observations allowed monitoring of the morphological and chemical modification of the materials during the different steps of functionalization. In vitro inorganic bioactivity was investigated by soaking samples in simulated body fluid. Enzymatic activity of the samples was tested and compared before and after soaking. Enzymatic activity of the solution was monitored at different experimental times. This study demonstrates that alkaline phosphatase could be successfully grafted onto different bioactive surfaces while maintaining its activity. Presence of the enzyme in vitro enhances the inorganic bioactivity of the materials tested.

Verné E; Ferraris S; Vitale-Brovarone C; Spriano S; Bianchi CL; Naldoni A; Morra M; Cassinelli C

2010-01-01

102

As fosfatases alcalinas, transaminases e gama-glutamil-transferase séricas em pacientes epilépticos tratados com carbamazepina/ The serum alkaline phosphatases, transaminases, and gamma-glutamil transferases in epileptic patients treated with carbamazepine  

Scientific Electronic Library Online (English)

Full Text Available Abstract in portuguese INTRODUÇÃO: A carbamazepina é a droga utilizada no tratamento de pacientes com epilepsia parcial (ou focal) secundariamente generalizada. Apesar do uso terapêutico, este fármaco tem sido implicado no aumento das atividades séricas de algumas enzimas. Alguns autores descreveram valores de prevalência de 7,7%, 13% e 22% para aumento de atividade das fosfatases alcalinas séricas (FA ou EC 3.1.3.1). A divergência de resultados também foi encontrada para as atividade (more) s da gama-glutamil-transferase sérica (gama-glutamil transferase ou GGT ou EC 2.3.2.2). OBJETIVO: Assim, a meta desta pesquisa é determinar, dentre outros objetivos, a freqüência de alterações nas atividades das FA, GGT e transaminases (AST, aspartato-amino-transferase, EC 2.6.1.1; e ALT, alanina-amino-transferase, EC 2.6.1.2) de uma amostra de pacientes do ambulatório de epilepsia em Salvador, Bahia. MATERIAL E MÉTODOS: O desenho do estudo é descritivo do tipo série de casos, aprovado pelo Comitê de Ética local, no qual uma amostra de conveniência de 52 pacientes epilépticos de acompanhamento ambulatorial foi obtida sem interferência dos pesquisadores. Estes pacientes foram organizados por faixa etária de 12 a 30 e de 31 a 90 anos e, subdivididos por tempo de monoterapia com carbamazepina. As atividades séricas das enzimas GGT, FA, AST e ALT foram determinadas. RESULTADOS: As proporções de alterações por variáveis foram descritas: 42% para as FA, 18% para as GGT, 2% para as ALT e 12% para as AST, respectivamente. A faixa etária de 12 a 30 anos apresentou 56% de alterações nas FA enquanto que aquela de 31 a 90 anos, apenas 18%. CONCLUSÃO: Nós concluímos que as enzimas FA, GGT, AST e ALT apresentaram maiores freqüências de alterações de suas atividades naqueles pacientes com idade igual ou inferior a 30 anos, sendo que as FA apresentaram maiores valores. Abstract in english INTRODUCTION: Carbamazepine is the drug of choice used in the treatment of patients with partial (or focal) epilepsy with secondary generalization. Despite its therapeutical use, this drug has been implicated in the increase of serum activities in some enzymes. Some authors have described prevalence values of 7.7%, 13%, and 22% for the increase of activity of serum alkaline phosphatases (AF or EC 3.1.3.1). A divergence in the results was also found for the activities of t (more) he serum g-glutamil transferase (gamma-glutamil transferase or GGT, or EC 2.3.2.2). OBJECTIVE: Hence, among other objectives, the aim of this research is to determine the frequency of sample alterations in serum enzymatic activities of AF, GGT and transaminases (AST, aspartate amino-transferase, EC 2.6.1.1; and ALT, alanine-amino-transferase, EC 2.6.1.2) in epilepsy ward patients in Salvador, Bahia. MATERIAL AND METHODS: The design of the study is descriptive and it is a case series type. It has been approved by the local Ethics Committee. In this study, a convenience sample of 52 epileptic patients who receive ambulatory care was obtained without interference by the researchers. These patients were divided according to age groups of 12 to 30 years and 31 to 90 years, which were then subdivided according to the period of monotherapy with carbamazepine. The serum activities of the enzymes GGT, AF, AST and ALT were determined. RESULTS: The ratios of alterations per variables were described: 42% for the FA, 18% for the GGT, 2% for the ALT, and 12% for the AST respectively. The age group of 12 to 30 years presented 56% of alterations in the AF while the group of 31 to 90 years presented only 18% of alterations. CONCLUSION: We conclude that the enzymes AF, GGT, AST, and ALT presented higher frequencies of alterations of their activities in those patients with age equal to or below 30 years, while the AF presented higher values.

Santos, Helder Jacobina; Andrade Filho, Antonio de Souza; Sanches, Olivia Lordelo; Spolador, Tiago; Rodrigues, Luís Erlon Araújo

2006-03-01

103

Study on alkaline and acid phosphatase activity in acute uranium intoxication  

International Nuclear Information System (INIS)

The protective potential of diethyl barbituric acid sodium salt is studied, in comparison with that of acetazolamide, on kidneys under acute uranium intoxication. Experiments involved rats given intraperitoneal injections with uranyl acetate on 12 successive days up to a total dose of 0.5, 2.0 or 7.0 mg/kg. The resulting effects are measured by chemical assays of serum and urine for alkaline and acid phosphatase and histochemical assays for phosphatase activities in kidneys, kinetics being followed over a 30-day period after total dose administration. Protection of kidneys from toxic uranium effects was found to be of about the same degree with sodium diethyl barbiturate as with acetazolamide. (A.B.)

1975-01-01

104

Effect of cobalt on synthesis and activation of Bacillus licheniformis alkaline phosphatase.  

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The effect of CO2+ on the synthesis and activation of Bacillus licheniformis MC14 alkaline phosphatase has been shown by the development of a defined minimal salts medium in which this organism produces 35 times more (assayable) alkaline phosphatase than when grown in a low-phosphate complex medium ...

Spencer, D B; Chen, C P; Hulett, F M

105

Identification of a Regulated Alkaline Phosphatase, a Cell Surface-Associated Lipoprotein, in Mycobacterium smegmatis  

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Although alkaline phosphatases are common in a wide variety of bacteria, there has been no prior evidence for alkaline phosphatases in Mycobacterium smegmatis. Here we report that transposon insertions in the pst operon, encoding homologues of an inorganic phosphate transporter, leads to constitutiv...

Kriakov, Jordan; Lee, Sun hee; Jacobs, William R.

106

Alkaline phosphatase-catalyzed silver deposition for electrochemical detection.  

Science.gov (United States)

Alkaline phosphatase (AP) is one of the most used enzymatic labels for the development of ELISAs, immunosensors, DNA hybridization assays, etc. This enzyme catalyzes the dephosphorylation of a substrate into a detectable product usually quantified by optical or electrochemical measurements. This work is based on a substrate (3-indoxyl phosphate) that produces a compound able to reduce silver ions in solution into a metallic deposit, which is localized where the enzymatic label AP is attached. The deposited silver is electrochemically stripped into solution and measured by anodic stripping voltammetry. Its application to an enzymatic genosensor on streptavidin-modified screen-printed carbon electrodes for the detection of virulence nucleic acid determinants of autolysin gene, exclusively present on the genome of the human pathogen Streptococcus pneumoniae, is described. Compared with the direct voltammetric detection of indigo carmine, the anodic stripping voltammetry of silver ions is 14-fold more sensitive. PMID:17569504

Fanjul-Bolado, Pablo; Hernández-Santos, David; González-García, María Begoña; Costa-García, Agustín

2007-06-15

107

Alkaline phosphatase-catalyzed silver deposition for electrochemical detection.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase (AP) is one of the most used enzymatic labels for the development of ELISAs, immunosensors, DNA hybridization assays, etc. This enzyme catalyzes the dephosphorylation of a substrate into a detectable product usually quantified by optical or electrochemical measurements. This work is based on a substrate (3-indoxyl phosphate) that produces a compound able to reduce silver ions in solution into a metallic deposit, which is localized where the enzymatic label AP is attached. The deposited silver is electrochemically stripped into solution and measured by anodic stripping voltammetry. Its application to an enzymatic genosensor on streptavidin-modified screen-printed carbon electrodes for the detection of virulence nucleic acid determinants of autolysin gene, exclusively present on the genome of the human pathogen Streptococcus pneumoniae, is described. Compared with the direct voltammetric detection of indigo carmine, the anodic stripping voltammetry of silver ions is 14-fold more sensitive.

Fanjul-Bolado P; Hernández-Santos D; González-García MB; Costa-García A

2007-07-01

108

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

109

[Cellular components and placental alkaline phosphatase in Trypanosoma cruzi infection ].  

UK PubMed Central (United Kingdom)

Trypanosoma cruzi induces changes in the protein pattern of human placenta syncytiotrophoblast. Placental alkaline phosphatase (PLAP) is a glycoenzyme anchored to the membrane by a glycosyl-phosphatidylinositol molecule. PLAP activity and its presence was altered by the parasite in cultures of human placental villi and HEp2 cells with T.cruzi. The cells treated before the cultures with agents which affect PILAP or glycosyl-phosphatidylinositol (antibodies, PL-C, genistein, lithium) presented less parasitic invasion than the control ones. It was also observed a modification in the pattern of actine filaments of the host cells infected. We concluded that PLAP would participate in the process of T. cruzi invasion into placental syncitiotrophoblast cells, by a mechanism that involves hydrolysis of the glycosyl-phosphatidylinositol molecules, the activation of tyrosine kinase proteins, the increase of cytosolic calcium and the rearrangement of actine filaments of the host cells.

Sartori MJ; Mezzano L; Lin S; Repossi G; Fabro SP

2005-01-01

110

Elevation of serum acid phosphatase in cancers with bone metastasis  

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In patients with nonprostatic cancer, serum acid phosphatase activity is usually elevated when bone metastasis is present but not when bone metastasis is absent. The fraction responsible for serum enzyme elevation is a normal component of serum; it appears in gel electrophoresis as band 5; and is tartrate-resistant. It is suggested that the origin of acid phosphatase elevation is bone osteoclasts rather than cancer tissue, as is the case with prostatic carcinoma. Determination of serum acid phosphatase activity may be useful in the detection of bone metastasis.

Tavassoli, M. (Scripps Clinic and Research Foundation, La Jolla, CA); Rizo, M.; Yam, L.T.

1980-05-01

111

Lack of relationship between activity of intestinal alkaline phosphatase and calcium or phosphate absorption.  

UK PubMed Central (United Kingdom)

The effects of vitamin D3 and the aqueous extract of Solanum malacoxylon on intestinal alkaline phosphatase and tissue phosphate content were studied on rachitic chicks treated with large doses of ethane-1-hydroxy-1,1 diphosphonate (EHDP). The EHDP treatment blocks the increase of intestinal calcium or phosphate absorption induced by the vitamin D3, while it has no effects on the rise of intestinal alkaline phosphatase activity or the increment in tissue phosphate content. The lack of correlation between the increment of alkaline phosphatase and that of Ca or phosphate absorption in vitamin D3 plus EHDP treated chicks excludes a participation of the alkaline phosphatase in the mechanism of Ca or P intestinal absorption. The Ca or phosphorus absorption are elicited specifically by 1,25-(OH)2-D3, while alkaline phosphatase activity and phosphate tissue concentration respond to a broader spectrum of stimuli.

Asteggiano C; Tolosa N; Pereira R; Moreno J; Cañas F

1981-01-01

112

Lack of relationship between activity of intestinal alkaline phosphatase and calcium or phosphate absorption.  

Science.gov (United States)

The effects of vitamin D3 and the aqueous extract of Solanum malacoxylon on intestinal alkaline phosphatase and tissue phosphate content were studied on rachitic chicks treated with large doses of ethane-1-hydroxy-1,1 diphosphonate (EHDP). The EHDP treatment blocks the increase of intestinal calcium or phosphate absorption induced by the vitamin D3, while it has no effects on the rise of intestinal alkaline phosphatase activity or the increment in tissue phosphate content. The lack of correlation between the increment of alkaline phosphatase and that of Ca or phosphate absorption in vitamin D3 plus EHDP treated chicks excludes a participation of the alkaline phosphatase in the mechanism of Ca or P intestinal absorption. The Ca or phosphorus absorption are elicited specifically by 1,25-(OH)2-D3, while alkaline phosphatase activity and phosphate tissue concentration respond to a broader spectrum of stimuli. PMID:6316731

Asteggiano, C; Tolosa, N; Pereira, R; Moreno, J; Cañas, F

1981-01-01

113

Alkaline phosphatase, leucine aminopeptidase, and alanine aminotransferase activities with obstructive and toxic hepatic disease in cats.  

UK PubMed Central (United Kingdom)

The activities of serum alkaline phosphatase (serum ALP), leucine aminopeptidase (serum LAP), and alanine aminotransferase (serum ALT) were determined in 15 cats before and after treatment by 3 methods: common bile duct occlusion, left hepatic duct(s) occlusion, and carbon tetrachloride administration. Significant increases in serum ALP, LAP, and ALT activities occurred in all cats in the 3 groups. Sustained mean increases of ninefold in ALP and 13-fold in LAP occurred in the cats with common bile duct occlusion. Lesser mean increases of these enzymes (fourfold) occurred in the cats with partial biliary occlusion. Transient mean increases (100-fold) in ALT occurred in the carbon tetrachloride-treated cats. Urine ALP excretion was measured in 3 cats with common bile duct occlusion. There was no significant difference between rates of urine ALP excretion before and after common bile duct occlusion. Specific ALP activities of hepatic extracts from normal cats and biliary-obstructed cats were compared. Mean specific activity was onefold higher in liver from cats with common bile duct occlusion of 21 days' duration. The findings in the present studies were interpreted to indicate that serum ALP and LAP are useful to detect biliary occlusive disease in cats and, in conjunction with serum ALT, may be used to differentiate primary hepatodegenerative disease and biliary occlusive disease.

Everett RM; Duncan JR; Prasse KW

1977-07-01

114

Total and bone-specific alkaline phosphatases in haemodialysis patients with chronic liver disease.  

UK PubMed Central (United Kingdom)

OBJECTIVE: The Kidney Disease: Improving Global Outcomes "KDIGO" recommends regular sampling of bone turnover markers (BTMs) such as total alkaline phosphatases (t-ALP) and bone-specific alkaline phosphatase (b-ALP) in the case of haemodialysis (HD) patients. DESIGN AND METHODS: We present our results of the regular assessment of t-ALP, b-ALP, and PTH, obtained for existing HD patients with chronic liver disease (LD). RESULTS: 76 prevalent HD patients were examined. Linear regression showed that b-ALP and t-ALP levels were closely related (r²: 0.6; p<0.0001), even when the serum PTH level was <250 pg/mL (r²: 0.56; p<0.001). The b-ALP/t-ALP ratio was 0.07 ± 0.12 and correlated poorly with PTH levels (r²: 0.03; p=0.01). Both b-ALP and t-ALP levels did not correlated with PTH levels. CONCLUSION: Our results did not confirm the KDIGO recommendation for using b-ALP as BTM in the special cases of HD patients with LDs.

Jean G; Souberbielle JC; Zaoui E; Lorriaux C; Mayor B; Hurot JM; Deleaval P; Chazot C

2012-04-01

115

How Should an Increase in Alkaline Phosphatase Activity Be Interpreted?  

International Nuclear Information System (INIS)

[en] 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

116

Nanoceria particles as catalytic amplifiers for alkaline phosphatase assays.  

UK PubMed Central (United Kingdom)

We propose a novel system to enhance detection sensitivity of alkaline phosphatase (ALP) by using nanoceria particles as redox active catalytic amplifiers of ALP signals in electrochemical assays. The catalytic activity of nanoceria particles attributed to their dual oxidation state Ce+4/Ce+3 and high oxygen mobility enhanced oxidation of the products of the ALP catalyzed reaction. A suite of spectroscopic and electrochemical methods including UV-VIS spectroscopy, transmission electron microscopy (TEM), dynamic light scattering (DLS) and cyclic voltammetry (CV) were used to characterize the interaction of nanoceria with the ALP generated products. Spectrometric experiments demonstrate change in the oxidation state of nanoceria upon exposure to the hydrolytic products of ALP. Three enzymatically generated products of commonly used ALP substrates were detected at a screen printing electrode surface in the presence of nanoceria. Electrochemical experiments demonstrate signal amplification of the ALP activity assay by nanoceria for all three products, demonstrating remarkable sensitivity of this assay. The assay was optimized with respect to pH and buffer composition. Analytical characterization of the nanoceria based ALP activity assay was established using 1-naphthyl phosphate substrate. The proposed strategy can find wide spread applications in sensing schemes involving ALP.

Hayat A; Andreescu S

2013-09-01

117

Bone Alkaline Phosphatase in CKD-Mineral Bone Disorder.  

Science.gov (United States)

Overall and cardiovascular mortality in patients with chronic kidney disease (CKD) is greatly increased, without obvious current effective treatments. Mineral and bone disorder (MBD) is a common manifestation of CKD and contributes to the high risk of fracture and cardiovascular mortality in these patients. Traditionally, clinical management of CKD-MBD focused on attenuation of secondary hyperparathyroidism due to impaired renal activation of vitamin D and phosphate retention, although recently, adynamic forms of renal bone disease have become more prevalent. Definitive diagnosis was based on histologic (histomorphometric) analysis of bone biopsy material supported by radiologic changes and changes in levels of surrogate laboratory markers. Of these various markers, parathyroid hormone (PTH) has been considered to be the most sensitive and currently is the most frequently used; however, the many pitfalls of measuring PTH in patients with CKD increasingly are appreciated. We propose an alternative or complementary approach using bone alkaline phosphatase (ALP), which is directly related to bone turnover, reflects bone histomorphometry, and predicts outcomes in hemodialysis patients. Here, we consider the overall merits of bone ALP as a marker of bone turnover in adults with CKD-MBD, examine published bone histomorphometric data comparing bone ALP to PTH, and discuss possible pathogenic mechanisms by which bone ALP may be linked to outcomes in patients with CKD. PMID:23623575

Sardiwal, Sunita; Magnusson, Per; Goldsmith, David J A; Lamb, Edmund J

2013-04-23

118

Relation of oxidative stress, zinc and alkaline phosphatase in protein energy malnutrition.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To determine serum zinc (Zn), total anti-oxidant capacity (TAC), malondialdehyde (MDA), alkaline phosphatase (ALP) and albumin in protein energy malnutrition (PEM) and to analyse the appropriateness of using low weight-for-age for detecting childhood under-nutrition. METHODS: This study comprised 455 children (355 malnourished and 100 normal). They were classified according to the Nutrition Subcommittee of Indian Academy of Pediatrics, Z-Score Classification and Composite Index of Anthropometric Failure. Serum Zn, TAC, MDA, ALP and albumin levels were determined. RESULTS: The serum Zn, TAC, ALP and albumin levels were found to be significantly decreased and MDA levels were significantly increased in malnourished children as compared with control (P < 0.001). CONCLUSION: The significant increase in serum MDA concentration associated with the decrease in serum TAC, Zn and ALP in malnourished children suggest that these children were potentially susceptible to high oxidative stress. Current study also suggest that conventional measures of detecting under-nutrition (low weight-for-age) may be missing out a considerable proportion of undernourished children present in the population.

Jain A; Jadhav AA; Varma M

2013-02-01

119

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

Energy Technology Data Exchange (ETDEWEB)

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 were largely dispersed, and only a few clusters were found. The gold-IgG binding was quantitatively estimated by stereological analysis of labeled, frozen thin sections. This estimation of a variety of control samples showed that the labeling was specific for the alkaline phosphatase. Cluster formation of the gold -IgG particles in association with the plasma membrane suggests that existence of specific alkaline phosphatase binding sites (receptors) in the plasma membrane of B. licheniformis 749/C. 27 references, 6 figures, 1 table.

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

1984-08-01

120

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

International Nuclear Information System (INIS)

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 were largely dispersed, and only a few clusters were found. The gold-IgG binding was quantitatively estimated by stereological analysis of labeled, frozen thin sections. This estimation of a variety of control samples showed that the labeling was specific for the alkaline phosphatase. Cluster formation of the gold -IgG particles in association with the plasma membrane suggests that existence of specific alkaline phosphatase binding sites (receptors) in the plasma membrane of B. licheniformis 749/C. 27 references, 6 figures, 1 table

1984-01-01

 
 
 
 
121

Alkaline phosphatase activity in plasma and liver of rats submitted to chronic exposure to fluoride  

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Full Text Available The aim of this study was to compare the effect of fluoride (F) on alkaline phosphatase activity in the liver and plasma of the rats. Four groups of male Wistar rats (n=6), which received drinking water containing 5, 15 or 50 ppm F or deionized water (control) throughout the experiment were included in the study. The animals were euthanized and had their tissues and blood plasma collected for the analysis of fluoride and alkaline phosphatase. There was an increase in F concentration in most tissues in the animals treated with higher F concentrations, except for the heart. The alkaline phosphatase assay showed an increase in the activity in the liver and blood plasma of the animals treated with fluoride concentrations of 15 and 50 ppm (p<0.05). This study suggested that F at a concentration of 50 ppm in drinking water promotes increased the activity of alkaline phosphatase in the liver and blood plasma.

Mileni da Silva Fernandes; Flávia Godoy Iano; Vivian Rocia; Marcela Mitsuko Yanai; Aline de Lima Leite; Tatiana Almeida Furlani; Marília Afonso Rabelo Buzalaf; Rodrigo Cardoso de Oliveira

2011-01-01

122

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

123

Polyoxometalates as potent and selective inhibitors of alkaline phosphatases with profound anticancer and amoebicidal activities.  

Science.gov (United States)

The biological significance of polyoxometalates is well renowned owing to their anticancer, antiviral and antibiotic properties. Here another therapeutic aspect of polyoxometalates has been explored as alkaline phosphatase inhibitors along with the remarked anticancer and amoebicidal properties. Synthesis and inhibitory studies of a set of seven polyoxotungstates against two major isozymes of alkaline phosphatase i.e. tissue specific and tissue non-specific alkaline phosphatase revealed their promising activity as alkaline phosphatase inhibitors. All compounds exhibited alkaline phosphatase inhibitory potency in nanomolar ranges. For tissue specific alkaline phosphatase, Na(10)[H(2)W(12)O(42)]·27H(2)O (A6) was found to be the most potent inhibitor (K(i) value 313 ± 7 nM), while for tissue non-specific alkaline phosphatase Na(33)[H(7)P(8)W(48)O(184)]·92H(2)O (A3) showed the highest inhibition potency (K(i) values 135 ± 10 nM). Moreover cytotoxicity evaluation of these compounds against lung carcinoma cells and immortalized human corneal epithelial cells demonstrated their anticancer potential with no cytotoxic effects on normal human cell lines. All anticancer drugs result in an impaired immune system and such immunocompromised persons become vulnerable to opportunistic infections specially Acanthamoeba which causes granulomatous amoebic encephalitis (GAE) which almost always results in death. The exclusive property of our tested polyoxotungstates is their strong amoebicidal activity against Acanthamoeba. Hence the study reveals a new window towards cancer therapy with the combined control of elevated levels of alkaline phosphatase and granulomatous amoebic encephalitis in cancer patients. PMID:23044850

Raza, Rabia; Matin, Abdul; Sarwar, Sundas; Barsukova-Stuckart, Maria; Ibrahim, Masooma; Kortz, Ulrich; Iqbal, Jamshed

2012-12-21

124

Polyoxometalates as potent and selective inhibitors of alkaline phosphatases with profound anticancer and amoebicidal activities.  

UK PubMed Central (United Kingdom)

The biological significance of polyoxometalates is well renowned owing to their anticancer, antiviral and antibiotic properties. Here another therapeutic aspect of polyoxometalates has been explored as alkaline phosphatase inhibitors along with the remarked anticancer and amoebicidal properties. Synthesis and inhibitory studies of a set of seven polyoxotungstates against two major isozymes of alkaline phosphatase i.e. tissue specific and tissue non-specific alkaline phosphatase revealed their promising activity as alkaline phosphatase inhibitors. All compounds exhibited alkaline phosphatase inhibitory potency in nanomolar ranges. For tissue specific alkaline phosphatase, Na(10)[H(2)W(12)O(42)]·27H(2)O (A6) was found to be the most potent inhibitor (K(i) value 313 ± 7 nM), while for tissue non-specific alkaline phosphatase Na(33)[H(7)P(8)W(48)O(184)]·92H(2)O (A3) showed the highest inhibition potency (K(i) values 135 ± 10 nM). Moreover cytotoxicity evaluation of these compounds against lung carcinoma cells and immortalized human corneal epithelial cells demonstrated their anticancer potential with no cytotoxic effects on normal human cell lines. All anticancer drugs result in an impaired immune system and such immunocompromised persons become vulnerable to opportunistic infections specially Acanthamoeba which causes granulomatous amoebic encephalitis (GAE) which almost always results in death. The exclusive property of our tested polyoxotungstates is their strong amoebicidal activity against Acanthamoeba. Hence the study reveals a new window towards cancer therapy with the combined control of elevated levels of alkaline phosphatase and granulomatous amoebic encephalitis in cancer patients.

Raza R; Matin A; Sarwar S; Barsukova-Stuckart M; Ibrahim M; Kortz U; Iqbal J

2012-12-01

125

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

Directory of Open Access Journals (Sweden)

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.

Joanne H. Wang; Kevin Wang; Brandon Bartling; Chung-Chiun Liu

2009-01-01

126

Modeling catalytic promiscuity in the alkaline phosphatase superfamily.  

Science.gov (United States)

In recent years, it has become increasingly clear that promiscuity plays a key role in the evolution of new enzyme function. This finding has helped to elucidate fundamental aspects of molecular evolution. While there has been extensive experimental work on enzyme promiscuity, computational modeling of the chemical details of such promiscuity has traditionally fallen behind the advances in experimental studies, not least due to the nearly prohibitive computational cost involved in examining multiple substrates with multiple potential mechanisms and binding modes in atomic detail with a reasonable degree of accuracy. However, recent advances in both computational methodologies and power have allowed us to reach a stage in the field where we can start to overcome this problem, and molecular simulations can now provide accurate and efficient descriptions of complex biological systems with substantially less computational cost. This has led to significant advances in our understanding of enzyme function and evolution in a broader sense. Here, we will discuss currently available computational approaches that can allow us to probe the underlying molecular basis for enzyme specificity and selectivity, discussing the inherent strengths and weaknesses of each approach. As a case study, we will discuss recent computational work on different members of the alkaline phosphatase superfamily (AP) using a range of different approaches, showing the complementary insights they have provided. We have selected this particular superfamily, as it poses a number of significant challenges for theory, ranging from the complexity of the actual reaction mechanisms involved to the reliable modeling of the catalytic metal centers, as well as the very large system sizes. We will demonstrate that, through current advances in methodologies, computational tools can provide significant insight into the molecular basis for catalytic promiscuity, and, therefore, in turn, the mechanisms of protein functional evolution. PMID:23728154

Duarte, Fernanda; Amrein, Beat Anton; Kamerlin, Shina Caroline Lynn

2013-06-03

127

Intestinal alkaline phosphatase inhibits the proinflammatory nucleotide uridine diphosphate.  

UK PubMed Central (United Kingdom)

Uridine diphosphate (UDP) is a proinflammatory nucleotide implicated in inflammatory bowel disease. Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor capable of inhibiting intestinal inflammation. We used the malachite green assay to show that IAP dephosphorylates UDP. To study the anti-inflammatory effect of IAP, UDP or other proinflammatory ligands (LPS, flagellin, Pam3Cys, or TNF-?) in the presence or absence of IAP were applied to cell cultures, and IL-8 was measured. UDP caused dose-dependent increase in IL-8 release by immune cells and two gut epithelial cell lines, and IAP treatment abrogated IL-8 release. Costimulation with UDP and other inflammatory ligands resulted in a synergistic increase in IL-8 release, which was prevented by IAP treatment. In vivo, UDP in the presence or absence of IAP was instilled into a small intestinal loop model in wild-type and IAP-knockout mice. Luminal contents were applied to cell culture, and cytokine levels were measured in culture supernatant and intestinal tissue. UDP-treated luminal contents induced more inflammation on target cells, with a greater inflammatory response to contents from IAP-KO mice treated with UDP than from WT mice. Additionally, UDP treatment increased TNF-? levels in intestinal tissue of IAP-KO mice, and cotreatment with IAP reduced inflammation to control levels. Taken together, these studies show that IAP prevents inflammation caused by UDP alone and in combination with other ligands, and the anti-inflammatory effect of IAP against UDP persists in mouse small intestine. The benefits of IAP in intestinal disease may be partly due to inhibition of the proinflammatory activity of UDP.

Moss AK; Hamarneh SR; Mohamed MM; Ramasamy S; Yammine H; Patel P; Kaliannan K; Alam SN; Muhammad N; Moaven O; Teshager A; Malo NS; Narisawa S; Millán JL; Warren HS; Hohmann E; Malo MS; Hodin RA

2013-03-01

128

Alkaline phosphatase and arterial structure and function in hypertensive African men: the SABPA study.  

UK PubMed Central (United Kingdom)

BACKGROUND: Vascular calcification is believed to be due to the conversion of vascular smooth muscle cells into osteoblast-like cells and is associated with mortality. Since hypertension and related mortality in Africans is a concern, we investigated associations between a marker of osteoblastic activity, alkaline phosphatase (ALP), and measures of arterial structure and function in hypertensive African men. METHODS: This study included 79 participants. We conducted 24h ambulatory blood pressure and carotid intima-media thickness (cIMT) measurements. cIMT was obtained with an intra-observer variability of 0.04 mm and the cross-sectional wall area (CSWA) was calculated. ALP was measured in serum. RESULTS: ALP was within its reference range (101.6 vs. 30.0-120.0 U/L), however cIMT was higher when this group was stratified and compared to gender and age-specific reference values. In univariate and partial regressions, and confirmed with multiple regression analyses, 24h systolic blood pressure (?=0.289, p=0.018), 24h pulse pressure (?=0.387, p=0.002), but not 24h diastolic blood pressure (?=0.073, p=0.58), were positively associated with ALP. In addition, mean cIMT (?=0.322, p=0.006) and CSWA (?=0.285, p=0.013) also correlated positively with ALP after adjusting for significant covariates, and after excluding participants with diabetes, renal dysfunction or a HIV positive status. CONCLUSION: Serum alkaline phosphatase is adversely associated with measures of arterial structure and function in hypertensive African men.

Schutte R; Huisman HW; Malan L; van Rooyen JM; Smith W; Glyn MC; Mels CM; Fourie CM; Malan NT; Schutte AE

2013-09-01

129

Identification of novel chromone based sulfonamides as highly potent and selective inhibitors of alkaline phosphatases.  

Science.gov (United States)

A new series of structurally diverse chromone containing sulfonamides has been developed. Crystal structures of three representative compounds (2a, 3a and 4a) in the series are reported. All compounds were screened for their inhibitory potential against alkaline phosphatases (ALPs). Two main classes of ALP isozymes were selected for this study, the tissue non-specific alkaline phosphatase (TNALP) from bovine and porcine source and the tissue-specific intestinal alkaline phosphatases (IALPs) from bovine source. All sulfonamide compounds had a marked preference for IALP (K(i), up to 0.01 ± 0.001 ?M) over TNALPs. Kinetics studies of the compounds showed competitive mode of inhibition. Molecular docking studies were carried out in order to characterize the selective inhibition of the compounds. An additional interesting aspect of these chromone sulfonamides is their inhibitory activity against ecto-5'-nucleotidase enzyme. PMID:23831808

al-Rashida, Mariya; Raza, Rabia; Abbas, Ghulam; Shah, Muhammad Shakil; Kostakis, George E; Lecka, Joanna; Sévigny, Jean; Muddassar, Muhammad; Papatriantafyllopoulou, Constantina; Iqbal, Jamshed

2013-06-19

130

Stability of an extreme halophilic alkaline phosphatase from Halobacterium salinarium in non-conventional medium.  

Science.gov (United States)

Alkaline p-nitrophenylphosphate phosphatase from the halophilic archaeon Halobacterium salinarum (earlier halobium) was solubilised in organic medium using reversed micelles of hexadecyltrimethylammonium bromide in cyclohexane, with 1-butanol as co-surfactant. The stability of alkaline p-nitrophenylphosphate phosphatase in this system was studied at different conditions, w(0) ([H(2)O]/[surfactant]), salt concentration, with and without Mn(+2). At all the conditions assayed, alkaline p-nitrophenylphosphate phosphatase was more stable in reversed micelles than in bulk aqueous solution (at 25 degrees C). The stabilisation effect of the reversed micelles was dramatic when the enzyme was dialysed against Mn(+2)-free buffer since the enzyme lost all the activity within 90 min in aqueous medium, but it retained approximately 72% of the initial enzymatic activity for 90 min in reversed micelles. PMID:11334667

Marhuenda-Egea, F C; Piera-Velázquez, S; Cadenas, C; Cadenas, E

2001-05-18

131

Stability of an extreme halophilic alkaline phosphatase from Halobacterium salinarium in non-conventional medium.  

UK PubMed Central (United Kingdom)

Alkaline p-nitrophenylphosphate phosphatase from the halophilic archaeon Halobacterium salinarum (earlier halobium) was solubilised in organic medium using reversed micelles of hexadecyltrimethylammonium bromide in cyclohexane, with 1-butanol as co-surfactant. The stability of alkaline p-nitrophenylphosphate phosphatase in this system was studied at different conditions, w(0) ([H(2)O]/[surfactant]), salt concentration, with and without Mn(+2). At all the conditions assayed, alkaline p-nitrophenylphosphate phosphatase was more stable in reversed micelles than in bulk aqueous solution (at 25 degrees C). The stabilisation effect of the reversed micelles was dramatic when the enzyme was dialysed against Mn(+2)-free buffer since the enzyme lost all the activity within 90 min in aqueous medium, but it retained approximately 72% of the initial enzymatic activity for 90 min in reversed micelles.

Marhuenda-Egea FC; Piera-Velázquez S; Cadenas C; Cadenas E

2001-05-01

132

Alkaline phosphatase activity in plasma and liver of rats submitted to chronic exposure to fluoride  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The aim of this study was to compare the effect of fluoride (F) on alkaline phosphatase activity in the liver and plasma of the rats. Four groups of male Wistar rats (n=6), which received drinking water containing 5, 15 or 50 ppm F or deionized water (control) throughout the experiment were included in the study. The animals were euthanized and had their tissues and blood plasma collected for the analysis of fluoride and alkaline phosphatase. There was an increase in F co (more) ncentration in most tissues in the animals treated with higher F concentrations, except for the heart. The alkaline phosphatase assay showed an increase in the activity in the liver and blood plasma of the animals treated with fluoride concentrations of 15 and 50 ppm (p

Fernandes, Mileni da Silva; Iano, Flávia Godoy; Rocia, Vivian; Yanai, Marcela Mitsuko; Leite, Aline de Lima; Furlani, Tatiana Almeida; Buzalaf, Marília Afonso Rabelo; Oliveira, Rodrigo Cardoso de

2011-12-01

133

Identification of novel chromone based sulfonamides as highly potent and selective inhibitors of alkaline phosphatases.  

UK PubMed Central (United Kingdom)

A new series of structurally diverse chromone containing sulfonamides has been developed. Crystal structures of three representative compounds (2a, 3a and 4a) in the series are reported. All compounds were screened for their inhibitory potential against alkaline phosphatases (ALPs). Two main classes of ALP isozymes were selected for this study, the tissue non-specific alkaline phosphatase (TNALP) from bovine and porcine source and the tissue-specific intestinal alkaline phosphatases (IALPs) from bovine source. All sulfonamide compounds had a marked preference for IALP (Ki, up to 0.01 ± 0.001 ?M) over TNALPs. Kinetics studies of the compounds showed competitive mode of inhibition. Molecular docking studies were carried out in order to characterize the selective inhibition of the compounds. An additional interesting aspect of these chromone sulfonamides is their inhibitory activity against ecto-5'-nucleotidase enzyme.

Al-Rashida M; Raza R; Abbas G; Shah MS; Kostakis GE; Lecka J; Sévigny J; Muddassar M; Papatriantafyllopoulou C; Iqbal J

2013-08-01

134

Bacteriophage PBSX-induced deletion mutants of Bacillus subtilis 168 constitutive for alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Mutants of Bacillus subtilis with deletions extending from the PBSX prophage, and in some cases removing pro(AB) and metC, have been found to be constitutive for vegetatively synthesized alkaline phosphatase. Such deletions were isolated by selecting for heat-resistant derivatives of a strain carrying a xhi-1479 mutation causing heat-inducibility of the defective phage PBSX. These deletions remove the phoS gene, a regulatory gene for alkaline phosphatase; it is concluded that the phoS gene product exerts negative control on alkaline phosphatase synthesis. Deletion mapping, combined with previously published linkage data, indicates a gene order of PBSX-phoS-pro(AB)-metC.

Piggot PJ; Buxton RS

1982-04-01

135

A Novel Hypothesis for an Alkaline Phosphatase 'Rescue' Mechanism in the Hepatic Acute Phase Immune Response.  

UK PubMed Central (United Kingdom)

The liver isoform of the enzyme alkaline phosphatase (AP) has been used classically as a serum biomarker for hepatic disease states such as hepatitis, steatosis, cirrhosis, drug-induced liver injury, and hepatocellular carcinoma. Recent studies have demonstrated a more general anti-inflammatory role for AP, as it is capable of dephosphorylating potentially deleterious molecules such as nucleotide phosphates, the pathogenic endotoxin lipopolysaccharide (LPS), and the contact clotting pathway activator polyphosphate (polyP), thereby reducing inflammation and coagulopathy systemically. Yet the mechanism underlying the observed increase in liver AP levels in circulation during inflammatory insults is largely unknown. This paper hypothesizes an immunological role for AP in the liver and the potential of this system for damping generalized inflammation along with a wide range of ancillary pathologies. Based on the provided framework, a mechanism is proposed in which AP undergoes transcytosis in hepatocytes from the canalicular membrane to the sinusoidal membrane during inflammation and the enzyme's expression is upregulated as a result. Through a tightly controlled, nucleotide-stimulated negative feedback process, AP is transported in this model as an immune complex with immunoglobulin G by the asialoglycoprotein receptor through the cell and secreted into the serum, likely using the receptor's State 1 pathway. The subsequent dephosphorylation of inflammatory stimuli by AP and uptake of the circulating immune complex by endothelial cells and macrophages may lead to decreased inflammation and coagulopathy while providing an early upstream signal for the induction of a number of anti-inflammatory gene products, including AP itself.

Pike AF; Kramer NI; Blaauboer BJ; Seinen W; Brands R

2013-07-01

136

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; P.V. Krupyanko; V.V. Belozerov

2005-01-01

137

Stereochemistry of phospho group transfer catalyzed by a mutant alkaline phosphatase  

International Nuclear Information System (INIS)

The stereochemical course of the phospho group transfer catalyzed by mutant (S102C) alkaline phosphatase from Escherichia coli was investigated by using 31P nuclear magnetic resonance spectroscopy. Transphosphorylation from 4-nitrophenyl (R/sub P/)-17O, 16O, 18O]phosphate to (S)-propane-1,2-diol occurs with overall retention of configuration at phosphorus. This result is consistent with the view that the hydrolysis of substrates by this mutant enzyme proceeds by way of a covalent phosphoenzyme intermediate in the same manner as the wild-type alkaline phosphatase.

1988-01-01

138

Stereochemistry of phospho group transfer catalyzed by a mutant alkaline phosphatase  

Energy Technology Data Exchange (ETDEWEB)

The stereochemical course of the phospho group transfer catalyzed by mutant (S102C) alkaline phosphatase from Escherichia coli was investigated by using /sup 31/P nuclear magnetic resonance spectroscopy. Transphosphorylation from 4-nitrophenyl (R/sub P/)-/sup 17/O, /sup 16/O, /sup 18/O)phosphate to (S)-propane-1,2-diol occurs with overall retention of configuration at phosphorus. This result is consistent with the view that the hydrolysis of substrates by this mutant enzyme proceeds by way of a covalent phosphoenzyme intermediate in the same manner as the wild-type alkaline phosphatase.

Butler-Ransohoff, J.E.; Kendall, D.A.; Freeman, S.; Knowles, J.R.; Kaiser, E.T.

1988-06-28

139

The Effect of Tamoxifen Treatment During 24 Months On Alkaline Phosphatase Concentration In Breast Cancer Patients  

Directory of Open Access Journals (Sweden)

Full Text Available Determination of alkaline phosphatase (ALP) isoenzyme activity is useful for the diagnosis and clinical evaluation of cancer patients, including breast cancer. 21 women were randomized to receive tamoxifen, probably the most widely used endocrine treatment for breast cancer worldwide, for 2 years, and 21 women constituted the control group. Control group were treated without Tamoxifen considering placebo effect. Alkaline phosphatase level was significantly lower in tamoxifen treated female breast cancer patients when compared with those of the treated patients with placebo. Results indicates that the tamoxifen decrease the level of ALP in first three months and then stabilize this level in the following months.

Betul Akcesme

2013-01-01

140

Inhibition of acid, alkaline, and tyrosine (PTP1B) phosphatases by novel vanadium complexes.  

UK PubMed Central (United Kingdom)

In the course of our investigations of vanadium-containing complexes for use as insulin-enhancing agents, we have generated a series of novel vanadium coordination complexes with bidentate ligands. Specifically we have focused on two ligands: anthranilate (anc(-)), a natural metabolite of tryptophan, and imidizole-4-carboxylate (imc(-)), meant to mimic naturally occurring N-donor ligands. For each ligand, we have generated a series of complexes containing the V(III), V(IV), and V(V) oxidation states. Each complex was investigated using phosphatase inhibition studies of three different phosphatases (acid, alkaline, and tyrosine (PTP1B) phosphatase) as prima facia evidence for potential use as an insulin-enhancing agent. Using p-nitrophenyl phosphate as an artificial phosphatase substrate, the levels of inhibition were determined by measuring the absorbance of the product at 405nm using UV/vis spectroscopy. Under our experimental conditions, for instance, V(imc)(3) appears to be as potent an inhibitor of alkaline phosphatase as sodium orthovanadate when comparing the K(cat)/K(m) term. VO(anc)(2) is as potent an inhibitor of acid phosphatase and tyrosine phosphatase as the Na(3)VO(4). Thus, use of these complexes can increase our mechanistic understanding of the effects of vanadium in vivo.

McLauchlan CC; Hooker JD; Jones MA; Dymon Z; Backhus EA; Greiner BA; Dorner NA; Youkhana MA; Manus LM

2010-03-01

 
 
 
 
141

Inhibition of acid, alkaline, and tyrosine (PTP1B) phosphatases by novel vanadium complexes.  

Science.gov (United States)

In the course of our investigations of vanadium-containing complexes for use as insulin-enhancing agents, we have generated a series of novel vanadium coordination complexes with bidentate ligands. Specifically we have focused on two ligands: anthranilate (anc(-)), a natural metabolite of tryptophan, and imidizole-4-carboxylate (imc(-)), meant to mimic naturally occurring N-donor ligands. For each ligand, we have generated a series of complexes containing the V(III), V(IV), and V(V) oxidation states. Each complex was investigated using phosphatase inhibition studies of three different phosphatases (acid, alkaline, and tyrosine (PTP1B) phosphatase) as prima facia evidence for potential use as an insulin-enhancing agent. Using p-nitrophenyl phosphate as an artificial phosphatase substrate, the levels of inhibition were determined by measuring the absorbance of the product at 405nm using UV/vis spectroscopy. Under our experimental conditions, for instance, V(imc)(3) appears to be as potent an inhibitor of alkaline phosphatase as sodium orthovanadate when comparing the K(cat)/K(m) term. VO(anc)(2) is as potent an inhibitor of acid phosphatase and tyrosine phosphatase as the Na(3)VO(4). Thus, use of these complexes can increase our mechanistic understanding of the effects of vanadium in vivo. PMID:20071031

McLauchlan, Craig C; Hooker, Jaqueline D; Jones, Marjorie A; Dymon, Zaneta; Backhus, Emily A; Greiner, Bradley A; Dorner, Nicole A; Youkhana, Mary A; Manus, Lisa M

2009-12-11

142

Activation and specificity of alkaline phosphatase of a mineralizing collagen-rich system.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase from tibia tendon of Meleagris gallopavo L. was highly purified. The enzyme activation by different ions was measured. Mg2+ showed a high activation with a broader spectrum of phosphomonoester hydrolization. The in vivo Mg2+ concentration was an optimum for in vitro activation.

Althoff J; Quint P; Höhling HJ

1978-06-01

143

Nucleotide sequence and characterization of the gene for secreted alkaline phosphatase from Lysobacter enzymogenes.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Lysobacter enzymogenes produces an alkaline phosphatase which is secreted into the medium. The gene for the enzyme (phoA) was isolated from a recombinant lambda library. It was identified within a 4.4-kb EcoRI-BamH1 fragment, and its sequence was determined by the chain termination method. The struc...

Au, S; Roy, K L; von Tigerstrom, R G

144

Purification and characterization of two extracellular alkaline phosphatases from a psychrophilic Arthrobacter isolate  

Energy Technology Data Exchange (ETDEWEB)

Two extracellular, heat-labile alkaline phosphatases were purified from a psychrophilic Arthrobacter isolate, D10. The enzymes were active over different pH ranges, used distinct substrates, and had different kinetic properties. Each enzyme reacted specifically to its own antibody during immunoblot analysis. One had both monopbosphatase and diesterase activities. 18 refs., 4 figs., 1 tab.

Prada, P. de; Brenchley, J.E. [Pennsylvania State Univ., University Park, PA (United States)

1997-07-01

145

Yeast permeabilization as a tool for measurement of in situ enzyme activity: localization of alkaline phosphatase.  

UK PubMed Central (United Kingdom)

The biochemical and ultracytochemical localization of alkaline phosphatases in permeabilized cells of Saccharomyces cerevisiae 257 has been studied. The treatment with non-ionic surfactant Triton X-100 allows the penetration of the substrate into intact yeast cells and thus provides detailed detection of the enzyme activity in ultracytochemical studies.

Spasova D; Galabova D

1998-05-01

146

Yeast permeabilization as a tool for measurement of in situ enzyme activity: localization of alkaline phosphatase.  

Science.gov (United States)

The biochemical and ultracytochemical localization of alkaline phosphatases in permeabilized cells of Saccharomyces cerevisiae 257 has been studied. The treatment with non-ionic surfactant Triton X-100 allows the penetration of the substrate into intact yeast cells and thus provides detailed detection of the enzyme activity in ultracytochemical studies. PMID:9679325

Spasova, D; Galabova, D

147

Fluorotyrosine Alkaline Phosphatase from Escherichia coli: Preparation, Properties, and Fluorine-19 Nuclear Magnetic Resonance Spectrum  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Alkaline phosphatase (EC 3.1.3.1) containing m-flurotyrosine has been prepared from E. coli grown in the presence of m-flurotyrosine. The kinetic properties of the m-fluorotyrosine enzyme measured with p-nitrophenylphosphate at pH 8.0 and dinitrophenylphosphate at pH 5.5 are essentially the same as ...

Sykes, Brian D.; Weingarten, Harold I.; Schlesinger, Milton J.

148

Fluorotyrosine Alkaline Phosphatase from Escherichia coli: Preparation, Properties, and Fluorine-19 Nuclear Magnetic Resonance Spectrum  

Science.gov (United States)

Alkaline phosphatase (EC 3.1.3.1) containing m-flurotyrosine has been prepared from E. coli grown in the presence of m-flurotyrosine. The kinetic properties of the m-fluorotyrosine enzyme measured with p-nitrophenylphosphate at pH 8.0 and dinitrophenylphosphate at pH 5.5 are essentially the same as those of normal alkaline phosphatase. However, the ability of the m-fluorotyrosine protein to refold active enzyme after acid denaturation, while unchanged at pH 5.8, was markedly decreased at pH 7.6. This result implies that the tyrosines must be in their protonated form for the protein to refold, reassociate, and take on zinc. The 19F nuclear magnetic resonance spectrum of m-fluorotyrosine alkaline phosphatase contains resolved resonances corresponding to different chemical environments for each m-fluorotyrosine in the folded protein. This demonstrates that 19F nuclear magnetic resonance spectroscopy of enzymes specifically labeled with 19F, even with enzymes as large as alkaline phosphatase (molecular weight, 86,000), will provide a very valuable probe for conformational changes in proteins. Images

Sykes, Brian D.; Weingarten, Harold I.; Schlesinger, Milton J.

1974-01-01

149

Rapid identification of Vibrio vulnificus on nonselective media with an alkaline phosphatase-labeled oligonucleotide probe.  

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An oligonucleotide DNA probe (VVAP) was constructed from a portion of the Vibrio vulnificus cytolysin gene (hylA) sequence and labeled with alkaline phosphatase covalently linked to the DNA. Control and environmental isolates probed with VVAP showed an exact correlation with results obtained with a ...

Wright, A C; Miceli, G A; Landry, W L; Christy, J B; Watkins, W D; Morris, J G

150

Optical Algal Biosensor using Alkaline Phosphatase for Determination of Heavy Metals  

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A biosensor is constructed to detect heavy metals from inhibition of alkaline phosphatase (AP) present on the external membrane of Chlorella vulgaris microalgae. The microalgal cells are immobilized on removable membranes placed in front of the tip of an optical fiber bundle inside a homemade microc...

Durrieu, Claude; Tran-Minh, Canh

151

Structural analysis of human adult and fetal alkaline phosphatases by cyanogen bromide peptide mapping.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The adult and fetal forms of human intestinal alkaline phosphatase (ALPase; orthophosphoric-monoester phosphohydrolase, EC 3.1.3.1) are indistinguishable by a variety of analytical procedures. However, they differ electrophoretically and can be differentiated by binding studies with monoclonal antib...

Vockley, J; D'Souza, M P; Foster, C J; Harris, H

152

Measurement of alkaline phosphatase of intestinal origin in plasma by p-bromotetramisole inhibition.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

L-p-bromotetramisole was used to inhibit non-intestinal alkaline phosphatase (of liver or bone origin) (EC 3.1.3.1; ALP) in plasma, and intestinal ALP was measured from the uninhibited activity. The method of determination is convenient and correlated well with measurement by immunocapture assay. If...

Kuwana, T; Rosalki, S B

153

Benign familial hyperphosphatasaemia as a cause of unexplained increase in plasma alkaline phosphatase activity.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

AIMS--To consider a possible genetic origin for the persistent unexplained increase in plasma alkaline phosphatase (ALP) in five non-related patients referred over an 18 month period. METHODS--Plasma ALP isoenzyme activities were measured in patients and their first degree relatives. RESULTS--In eac...

Rosalki, S B; Foo, A Y; Dooley, J S

154

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

Wang, Joanne H.; Wang, Kevin; Bartling, Brandon; Liu, Chung-Chiun

155

Alkaline phosphatase induces the mineralization of sheets of collagen implanted subcutaneously in the rat.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To determine whether alkaline phosphatase (ALP) can cause the mineralization of collagenous matrices in vivo, bovine intestinal ALP was covalently bound to slices of guanidine-extracted demineralized bovine dentin (DDS). The preparations were implanted subcutaneously over the right half of the rat s...

Beertsen, W; van den Bos, T

156

Pyrophosphate-regulated Zn(2+)-dependent DNAzyme activity: an amplified fluorescence sensing strategy for alkaline phosphatase.  

Science.gov (United States)

In this work, based on the fact that pyrophosphate (PPi) could regulate the activity of Zn(2+)-dependent DNAzyme, we for the first time report a fluorescence turn-on sensing system for alkaline phosphatase (ALP) with improved sensitivity via nonprotein-enzymatic signal amplification. A catalytic and molecular beacon (CAMB) design was employed to further improve its sensitivity. Taking advantage of the strong interactions between PPi and the Zn(2+), the cofactor Zn(2+) was caged, and the DNAzyme activity was effectively inhibited. The introduction of ALP, however, could catalyze the hydrolysis of PPi and release free Zn(2+), resulting in the activation of DNAzyme to catalyze the cleavage of the molecular beacon substrate with a remarkable increase of fluorescent signal. These optimized designs together allow a high sensitivity for ALP, with a detection limit of 20 pM observed, much lower than previously reported methods. It has also been used for detection of ALP in human serum with satisfactory results, demonstrating its potential applications in clinical diagnosis. PMID:23891797

Kong, Rong-Mei; Fu, Ting; Sun, Ni-Na; Qu, Feng-Li; Zhang, Shu-Fang; Zhang, Xiao-Bing

2013-07-05

157

A gold nanoparticles-based colorimetric assay for alkaline phosphatase detection with tunable dynamic range.  

UK PubMed Central (United Kingdom)

In this report, a simple and label-free colorimetric assay was developed for detecting alkaline phosphatase (ALP). Based on the conjugated gold nanoparticle/adenosine triphosphate (AuNP/ATP) sensing system, this assay is highly sensitive and selective. In this system, ATP induces the aggregation of cetyltrimethylammonium bromide (CTAB)-capped AuNPs and ALP stimulates the disaggregation of AuNPs by converting ATP into adenosine through an enzymatic dephosphorylation reaction. Hence, the presence of ALP can be visually observed (gray-to-red color change) and monitored by the shift of the surface plasmon resonance (SPR) absorption band of AuNPs. Furthermore, the dynamic range of the method can be varied by addition of different metal ions (e.g. 100-600unit/L to 5.0-100unit/L and 0.2-20unit/L in the presence of Ca(2+) and Pb(2+), respectively). The feasibility of this sensitive and specific assay with a tunable dynamic range was demonstrated to be consistent even in human serum samples.

Li CM; Zhen SJ; Wang J; Li YF; Huang CZ

2013-05-01

158

Alkaline phosphatase ALPPL-2 is a novel pancreatic carcinoma-associated protein.  

UK PubMed Central (United Kingdom)

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy with a very low median survival rate. The lack of early sensitive diagnostic markers is one of the main causes of PDAC-associated lethality. Therefore, to identify novel pancreatic cancer biomarkers that can facilitate early diagnosis and also help in the development of effective therapeutics, we developed RNA aptamers targeting pancreatic cancer by Cell-systematic evolution of ligands by exponential enrichment (SELEX) approach. Using a selection strategy that could generate aptamers for 2 pancreatic cancer cell lines in one selection scheme, we identified an aptamer SQ-2 that could recognize pancreatic cancer cells with high specificity. Next, by applying 2 alternative approaches: (i) aptamer-based target pull-down and (ii) genome-wide microarray-based identification of differentially expressed mRNAs in aptamer-positive and -negative cells, we identified alkaline phosphatase placental-like 2 (ALPPL-2), an oncofetal protein, as the target of SQ-2. ALPPL-2 was found to be ectopically expressed in many pancreatic cancer cell lines at both mRNA and protein levels. RNA interference-mediated ALPPL-2 knockdown identified novel tumor-associated functions of this protein in pancreatic cancer cell growth and invasion. In addition, the aptamer-mediated identification of ALPPL-2 on the cell surface and cell secretions of pancreatic cancer cells supports its potential use in the serum- and membrane-based diagnosis of PDAC.

Dua P; Kang HS; Hong SM; Tsao MS; Kim S; Lee DK

2013-03-01

159

EXPRESSION OF ALKALINE PHOSPHATASE DURING OSTEOGENIC DIFFERENTIATION OF RAT BONE MARROW STROMAL CELLS  

Directory of Open Access Journals (Sweden)

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

AKBARI M; NIKBAKHT M; SOBHANI A.GH

2001-01-01

160

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.

Fenton Tanis R; Lyon Andrew W; Rose M Sarah

2011-01-01

 
 
 
 
161

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

Science.gov (United States)

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

162

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

UK PubMed Central (United Kingdom)

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.

Hua G; Zhang R; Bayyareddy K; Adang MJ

2009-10-01

163

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.

M Salehnia; A Al Yassin; M Agha Hosseini; L Safdarian; A Khademi; H Saidi; Z Rezaieyan; Sh Byranvand

2006-01-01

164

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

UK PubMed Central (United Kingdom)

BACKGROUND AND AIMS: Intestinal alkaline phosphatase (IAP) is a gut mucosal defense factor known to dephosphorylate lipopolysaccharide (LPS); however, the role of IAP in the gut response to luminal bacteria remains poorly defined. We investigated immune responses of wild-type (WT) and IAP-knockout (IAP-KO) mice to LPS and Salmonella typhimurium challenges. METHODS: Cryostat sectioning and standard indirect immunohistochemical staining for major histocompatibility complex (MHC) class II molecules were performed on liver tissue from WT and IAP-KO mice. WT and IAP-KO mice were orally gavaged with S. typhimurium; bacterial translocation to mesenteric nodes, liver, and spleen was determined by tissue homogenization and plating. In other experiments, WT and IAP-KO mice received intraperitoneal injections of LPS, with subsequent quantification of complete blood counts and serum interleukin (IL)-6 by enzyme-linked immunosorbent assay (ELISA). WT and IAP-KO whole blood were plated and stimulated with LPS and Pam-3-Cys, followed by cytokine assays. RESULTS: Immunohistologic liver examinations showed increased expression of MHC class II molecules in IAP-KO mice. Following S. typhimurium challenge, WT mice appeared moribund compared with IAP-KO mice, with increased bacterial translocation. WT mice had >50% decrease (P<.005) in platelets and 1.8-fold (P<.05) increased serum IL-6 compared with IAP-KO mice in response to LPS injections. IAP-KO whole-blood stimulation with LPS and Pam-3-Cys resulted in increased IL-6 and tumor necrosis factor (TNF)-alpha secretion compared with WT. CONCLUSIONS: IAP-KO mice exhibit characteristics consistent with local LPS tolerance. Whole-blood response of IAP-KO mice did not reflect systemic tolerance. These data suggest that IAP is a local immunomodulating factor, perhaps regulating LPS-toll-like receptor 4 (TLR4) interaction between commensal microflora and intestinal epithelium.

Chen KT; Malo MS; Beasley-Topliffe LK; Poelstra K; Millan JL; Mostafa G; Alam SN; Ramasamy S; Warren HS; Hohmann EL; Hodin RA

2011-04-01

165

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

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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

166

Light microscope and electron microscope alkaline phosphatase cytochemistry of rat bone marrow leukocytes.  

UK PubMed Central (United Kingdom)

An enzyme cytochemical method yielding an osmiophilic reaction product, visible at both the light and electron microscope levels, has been applied to the study of alkaline phosphatase in rat bone marrow cells. The enzyme is present in both eosinophils and, in much smaller amounts, in neutrophils. In both cases it is present on the plasma membrane, and in eosinophils intracellular aggregations of reaction product are also seen. The specific granules in both cell types fail to react and the enzyme is first detectable at the promyelocyte stage. Thus the enzyme is demonstrable before specific granule formation begins in the neutrophil, indicating that they are not a significant site of alkaline phosphatase activity in the rat.

Williams DM; Gillett R; Linder JE

1979-02-01

167

Differential release of membrane-bound alkaline phosphatase isoenzymes from tumor cells by bromelain.  

UK PubMed Central (United Kingdom)

A rapid enzymatic method is presented which results in the selective release of cell-surface alkaline phosphatase isoenzymes. Treatment of suspensions of human tumor cell lines with the proteolytic enzyme bromelain released certain alkaline phosphatase isoenzymes into low-molecular-weight, catalytically active forms. Cells which expressed term placental or intestinal isoenzymes were equally susceptible to this treatment. A cell line which expressed early placental isoenzyme, however, was unaffected by bromelain as indicated by complete recovery of activity in the treated membrane fraction. Successful solubilization of immunologically reactive enzyme allows quantitation of levels of cell-surface enzyme in response to modulators of gene expression. Moreover, the observed selective solubilization of isoenzymes by bromelain may be of general use in analyses of the physical association between other biologically important surface proteins and the lipid components of the cell membrane.

Kottel RH; Hanford WC

1980-06-01

168

Distribution of placental alkaline phosphatase (PLAP) gene frequencies in Andhra Pradesh (south India).  

UK PubMed Central (United Kingdom)

The data available on the distribution of variants, phenotype and gene frequencies of different placental isoenzymes, especially for caste populations of Andhra Pradesh, are very scanty. In the present study, phenotype and gene frequency distributions of placental alkaline phosphatase (PLAP) of two caste groups namely, Lambada and Perika of Andhra Pradesh, South India, are reported. All the common phenotypes and the single case of a rare phenotype have been observed in the Lambada group.

Swamy TV; Kumar NS

1996-08-01

169

Bio-nanocapsules for signal enhancement of alkaline phosphatase-linked immunosorbent assays.  

UK PubMed Central (United Kingdom)

The bio-nanocapsules displaying about 240 molecules of immunoglobulin G Fc-binding Z domains (ZZ-BNCs) enhanced the signals of enzyme-linked immunosorbent assay by tethering the Fc regions of secondary antibodies (Abs), which were eliminated using high-molecular mass enzymes (e.g., alkaline phosphatase). By way of optimizing the distance between enzymes and Abs, ZZ-BNCs improved sensitivity independently of enzymes.

Iijima M; Yamamoto M; Yoshimoto N; Niimi T; Kuroda S

2013-01-01

170

Absent or extremely low neutrophil alkaline phosphatase activity levels in patients with myelodysplastic syndromes.  

Science.gov (United States)

Three patients with myelodysplastic syndrome (MDS) had absent or extremely low levels of neutrophil alkaline phosphatase (NAP) activity (arbitrarily defined as an NAP score <10). All patients showed varying degrees of hypogranulation in neutrophil morphology. The NAP activity levels transiently normalized following the administration of granulocyte colony-stimulating factor (G-CSF) in two cases. No patients experienced any severe infectious episodes. These results suggest that NAP activity is not central to the neutrophil function. PMID:23411705

Yoshida, Yataro; Katsurada, Tatsuya; Oguma, Shigeru; Nakabo, Yukiharu; Yoshinaga, Noriyoshi; Kawahara, Masahiro; Kawabata, Hiroshi

2013-02-15

171

Absent or extremely low neutrophil alkaline phosphatase activity levels in patients with myelodysplastic syndromes.  

UK PubMed Central (United Kingdom)

Three patients with myelodysplastic syndrome (MDS) had absent or extremely low levels of neutrophil alkaline phosphatase (NAP) activity (arbitrarily defined as an NAP score <10). All patients showed varying degrees of hypogranulation in neutrophil morphology. The NAP activity levels transiently normalized following the administration of granulocyte colony-stimulating factor (G-CSF) in two cases. No patients experienced any severe infectious episodes. These results suggest that NAP activity is not central to the neutrophil function.

Yoshida Y; Katsurada T; Oguma S; Nakabo Y; Yoshinaga N; Kawahara M; Kawabata H

2013-01-01

172

Studies on Excessive Daytime Sleepiness (EDS) and Alkaline Phosphatase Activity in Smoker and Alcoholic Person  

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Full Text Available Normal 0 false false false EN-US X-NONE X-NONE /* 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-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin-top:0in; mso-para-margin-right:0in; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0in; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} The alkaline phosphatase activity in saliva may be related to the excessive daytime sleepiness (EDS) to alcoholic, smoker and healthy subjects. 29 Saliva sample was collected from alcoholic categories, 22 from normal healthy and 11 from smoker categories. Estimation of level of enzyme activity (alkaline phosphatase) was identified by DGKC KIT.  Alkaline phosphatase activity was more observe in alcoholic, smoker than healthy person. 90% Alcoholic, 40% smoker and 20% healthy person were EDS.

Jayant Kumar; Shashi Shekhar Kumar; R K Pradhan

2011-01-01

173

Association between reduced levels of alkaline phosphatase and survival times of patients with primary sclerosing cholangitis.  

UK PubMed Central (United Kingdom)

BACKGROUND & AIMS: Ursodeoxycholic acid (UDCA) has not been shown to stop progression of primary sclerosing cholangitis (PSC). However, patients with primary biliary cirrhosis treated with UDCA whose levels of alkaline phosphatase (ALP) decrease have longer survival times than patients whose levels do not decrease. We compared survival times between patients with PSC treated with UDCA or placebo, with and without decreased levels of ALP. METHODS: We collected data from patients enrolled in the Scandinavian PSC UDCA trial. Patients were randomly assigned to groups given UDCA (17-23 mg/kg/day, n = 97) or placebo (n = 101) from 1996-2001 and were followed until 2010. End points were death, liver transplantation, or cholangiocarcinoma. They were considered to be biochemical responders if they had serum levels of ALP that were normal or reduced by ?40% after 1 year in the trial (regardless of whether they received UDCA or placebo). Numbers of patients surviving until the study end point were compared by using the Kaplan-Meier method. RESULTS: There were no differences in survival at the end of the study between patients given UDCA or placebo (P = .774, log-rank); 26 patients in the UDCA group and 29 in the placebo group reached an end point. On the basis of ALP levels, there were 79 responders and 116 nonresponders overall. Of patients given UDCA, significantly more biochemical responders survived for 10 years than nonresponders (P = .03, log-rank). However, differences remained significant regardless of group assignment; overall, patients with reductions in ALP level survived longer than patients without reductions in ALP (P = .0001, log-rank). CONCLUSIONS: There is no significant difference in long-term survival between patients with PSC given UDCA (17-23 mg/kg/day) or placebo for 5 years. However, patients who have reduced or normal levels of ALP have longer survival times, regardless of whether they receive UDCA or placebo.

Lindström L; Hultcrantz R; Boberg KM; Friis-Liby I; Bergquist A

2013-07-01

174

Immunolocalisation of testicular tumor using radiolabelled monoclonal antibody to placental alkaline phosphatase  

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Tumour associated monoclonal antibody against placental alkaline phosphatase (H17E2) was radiolabelled vith Indium-111 and Iodine-123 and administered intravenously in 33 patients with primary and/or metastatic testicular tumour, as well as in 8 patients who were in complete remission after surgical excision of the tumour. The presence of a tumour was confirmed and correlated well with conventional diagnostic techniques and, in addition, the antibody scan revealed the presence of active disease in 2 patients with negative conventional imaging and with elevated serum markers. In addition, in one patient the CD produced a false positive result where the antibody scan was negative. Finally, the absence of tumour was confirmed in all 8 cases of patients in complete remission. All patients studied with Indium-labelled antibody had odservable concentrations of the radiolabel in the liver (estimated to be approximately 30% of the administred dose), as well as in the kidneys and spleen. The patients studied with the Iodine-123 labelled antibody had odservable concentrations in the thyroid gland and the stomach. The best images were seen at 48 and 24 hrs after the Indium and Iodine radiolabelled antibody respectively. No human anti-mouse antibody was detected in any of our patients, even in those who received 2 and 3 administrations, with the highest amount of administred protein being 800 ?g. No toxicity was encountered in any of our patients in 4 months of follow-up. This method may be of clinical value in patients with testicular neoplasms and represents a new addition to current imaging techniques. A positive scan indicates the definite presence of a tumour. Antibody scans can contribute to the staging and long-term monitoring of patients for the presence of recurrent testicular tumours. A prospective study should be performed in order to define the overall sensitivity and specificity of this method

1990-01-01

175

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

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Full Text Available Ten duodeni of adult ram were fixed in chilled acetone, 80% ethyl alcohol, formol- alcohol solution, alcoholic bouinssolution and neutral buffered formalin solution. The distribution of alkaline and acid phosphatases were similar in theirlocation but different in their intensity and distribution according to different fixative The distribution of alkaline phosphatasein absorptive columnar cell was more intense than in goblet cells, whereas the concentration of acid phosphatase was moreintense in goblet cells than in absorptive cells in the mucosa of sheep duodenum. The study revealed that the samples wasfixed with chilled acetone gave highest reaction for alkaline and acid phosphatases than other fixative samples. No reaction foralkaline phosphatase include the lower parts of intestinal glands, paneth cells and sub mucosal glands in different fixative,whereas, paneth cells and sub mucosal glands revealed wreaked reaction for acid phosphatase in samples fixed in 80% ethylalcohol and chilled acetone respectively in duodenum of native sheep.

N. S. Ahmed

2010-01-01

176

Histochemical localization of alkaline phosphatase in the uterus of rats: response to a few indigenous plant extracts.  

Science.gov (United States)

Localization of alkaline phosphatase in the uterine luminal and glandular epithelium of rats under the influence of 50% ethanolic and benzene extracts of three indigenous plants viz. Embelia ribes Burm. (dried berries), Artobotrys odoratissimus Linn. (fresh green leaves) and Hibiscus rosasinensis Linn. (flowers) has been studied histochemically. 75 and 150 mg/kg doses of 50% ethanolic extracts of E. ribes increased the intensity of reaction for alkaline phosphatase in both luminal and glandular epithelium, whereas extracts of A. odorantissimus and H. rosa-sinensis could not elicit any significant positive staining in luminal and glandular epithelium for alkaline phosphatase. Intense positive reaction for alkaline phosphatase due to E. ribes extract has been correlated with its estrogenic mode of action. PMID:3577595

Mathur, R

177

Histochemical localization of alkaline phosphatase in the uterus of rats: response to a few indigenous plant extracts.  

UK PubMed Central (United Kingdom)

Localization of alkaline phosphatase in the uterine luminal and glandular epithelium of rats under the influence of 50% ethanolic and benzene extracts of three indigenous plants viz. Embelia ribes Burm. (dried berries), Artobotrys odoratissimus Linn. (fresh green leaves) and Hibiscus rosasinensis Linn. (flowers) has been studied histochemically. 75 and 150 mg/kg doses of 50% ethanolic extracts of E. ribes increased the intensity of reaction for alkaline phosphatase in both luminal and glandular epithelium, whereas extracts of A. odorantissimus and H. rosa-sinensis could not elicit any significant positive staining in luminal and glandular epithelium for alkaline phosphatase. Intense positive reaction for alkaline phosphatase due to E. ribes extract has been correlated with its estrogenic mode of action.

Mathur R

1986-07-01

178

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

UK PubMed Central (United Kingdom)

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

Yan Y; Peng L; Liu WX; Wan FH; Harris MK

2011-01-01

179

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

Science.gov (United States)

Bemisia tabaci (Gennadius) B-biotype and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) often coexist on greenhouse-grown vegetable crops in northern China. The recent spread of B. tabaci B-biotype has largely replaced T. vaporariorum, and B-biotype now overlaps with T. vaporariorum where common hosts occur in most invaded areas. The impact of the B-biotype on the agro eco system appears to be widespread, and involves the ability to compete with and perhaps replace other phytophages like T. vaporariorum. An emerging hypothesis is that the B-biotype is physiologically superior due at least in part to an improved ability to metabolically utilize the alkaline phosphatase pathway. To test this hypothesis, alkaline phosphatase activity was studied in the B-biotype and T. vaporariorum after feeding on a number of different hosts for a range of durations, with and without host switching. Alkaline phosphatase activity in T. vaporariorum was 1.45 to 2.53-fold higher than that of the B-biotype when fed on tomato for 4 and 24 h, or switched from tomato to cotton and cabbage for the same durations. However, alkaline phosphatase activity in the B-biotype was 1.40 to 3.35-fold higher than that of T. vaporariorum when the host switching time was ?72 and ?120 h on the same plant. Both short-term (4 h) and long-term (72 h) switching of plant hosts can significantly affect the alkaline phosphatase activity in the two species. After ?120 h, feeding on tomato and cotton alkaline phosphatase activity in the B-biotype was significantly higher than that of T. vaporariorum. It was shown that alkaline phosphatase aids the species feeding on different plant species, and that the B-biotype is physiologically superior to T. vaporariorum in utilizing the enzyme compared to T. vaporariorum over longer periods of feeding. PMID:21521136

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

2011-01-01

180

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 (pSEAP2-1)creating 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; Mohammad Ali Eghbal

2006-01-01

 
 
 
 
181

Cloning and characterization of the Cry1Ac-binding alkaline phosphatase (HvALP) from Heliothis virescens.  

UK PubMed Central (United Kingdom)

Membrane-bound alkaline phosphatases (mALPs, EC 3.1.3.1) in the insect midgut have been reported as functional receptors for Cry toxins from the bacterium Bacillus thuringiensis. We previously reported the identification of HvALP in the midgut of Heliothis virescens larvae as a Cry1Ac-binding protein that is down-regulated in Cry1Ac-resistant insects. To further characterize HvALP, we localized mALP protein to foregut and midgut tissues using anti-mALP serum and then cloned five mALPs from H. virescens larval midgut. All five clones displayed high levels of sequence identity (above 90%), suggesting that they may represent allelic variants, and grouped with other lepidopteran mALPs in sequence alignments. All these cloned ALPs were predicted to contain a glycosylphosphatidylinositol (GPI) anchor and were named HvmALP1-5. We expressed two of the most diverse HvmALPs in a heterologous system to test binding of Cry1Ac and recognition by HvALP cross-reacting antiserum. Our data highlight the importance of glycosylation for Cry1Ac binding to HvALP and suggest that, depending on glycosylation, all the identified HvmALPs may be synonymous with HvALP, the Cry1Ac-binding phosphatase identified in H. virescens midgut epithelium.

Perera OP; Willis JD; Adang MJ; Jurat-Fuentes JL

2009-04-01

182

Cloning and characterization of the Cry1Ac-binding alkaline phosphatase (HvALP) from Heliothis virescens.  

Science.gov (United States)

Membrane-bound alkaline phosphatases (mALPs, EC 3.1.3.1) in the insect midgut have been reported as functional receptors for Cry toxins from the bacterium Bacillus thuringiensis. We previously reported the identification of HvALP in the midgut of Heliothis virescens larvae as a Cry1Ac-binding protein that is down-regulated in Cry1Ac-resistant insects. To further characterize HvALP, we localized mALP protein to foregut and midgut tissues using anti-mALP serum and then cloned five mALPs from H. virescens larval midgut. All five clones displayed high levels of sequence identity (above 90%), suggesting that they may represent allelic variants, and grouped with other lepidopteran mALPs in sequence alignments. All these cloned ALPs were predicted to contain a glycosylphosphatidylinositol (GPI) anchor and were named HvmALP1-5. We expressed two of the most diverse HvmALPs in a heterologous system to test binding of Cry1Ac and recognition by HvALP cross-reacting antiserum. Our data highlight the importance of glycosylation for Cry1Ac binding to HvALP and suggest that, depending on glycosylation, all the identified HvmALPs may be synonymous with HvALP, the Cry1Ac-binding phosphatase identified in H. virescens midgut epithelium. PMID:19552892

Perera, Omaththage P; Willis, Jonathan D; Adang, Michael J; Jurat-Fuentes, Juan L

2009-02-07

183

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

184

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

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Full Text Available 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 group. FT-IR confirmed no decomposition in pyranose ring in P-COS even with heating and treatment in acidic conditions. The amorphous nature of P-COS was confirmed by X-ray diffraction analysis. Further, the biocompatibility and alkaline phosphatase activity of P-COS were checked against the osteosarcoma MG63 cell line at different concentrations and no cytotoxicity was observed. After 12 h and 24 h of incubation, the ALP activity of P-COS was higher compared with the control group. These results suggest that P-COS is a biocompatible material and in future P-COS could open up a number of promising pharmaceutical and clinical applications to mankind.

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

2010-01-01

185

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

UK PubMed Central (United Kingdom)

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).

Giraudet H; Berthon JL; Buisson B

1997-06-01

186

[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

187

Implication of BBM lipid composition and fluidity in mitigated alkaline phosphatase activity in renal cell carcinoma.  

Science.gov (United States)

Previous study has documented reduced alkaline phosphatase (ALP) activity in brush border membrane (BBM) isolated from renal cell carcinoma (RCC). Diminished activity of ALP is associated with alteration in both increased K(m) as well as decreased V(max) of enzyme suggests that there may be a change in the conformation of enzyme as well as decreased number of ALP active molecules. The present study was conducted to find out any role of BBM lipid composition and its fluidity in diminished activity of alkaline phosphatase in renal cell carcinoma. Total phospholipids and glycolipids were significantly augmented in BBM from RCC as compared to control. Fractional analysis of total phospholipids revealed significantly increased phosphatidylethanolamine. Decreased fractions of sphingomyelin and phosphatidylinositol were observed. Cholesterol-to-total phospholipid molar ratios in tumor BBM was a significantly lower in tumor BBM. A significant reduction in polarization and microviscosity was found in BBM from RCC. Therefore, we conclude that alteration in membrane lipid composition and fluidity may play a substantial role in reduced activity of ALP in RCC. PMID:22810501

Sharma, Ujjawal; Singh, Shrawan Kumar; Pal, Deeksha; Khajuria, Ragini; Mandal, Arup Kumar; Prasad, Rajendra

2012-07-19

188

Implication of BBM lipid composition and fluidity in mitigated alkaline phosphatase activity in renal cell carcinoma.  

UK PubMed Central (United Kingdom)

Previous study has documented reduced alkaline phosphatase (ALP) activity in brush border membrane (BBM) isolated from renal cell carcinoma (RCC). Diminished activity of ALP is associated with alteration in both increased K(m) as well as decreased V(max) of enzyme suggests that there may be a change in the conformation of enzyme as well as decreased number of ALP active molecules. The present study was conducted to find out any role of BBM lipid composition and its fluidity in diminished activity of alkaline phosphatase in renal cell carcinoma. Total phospholipids and glycolipids were significantly augmented in BBM from RCC as compared to control. Fractional analysis of total phospholipids revealed significantly increased phosphatidylethanolamine. Decreased fractions of sphingomyelin and phosphatidylinositol were observed. Cholesterol-to-total phospholipid molar ratios in tumor BBM was a significantly lower in tumor BBM. A significant reduction in polarization and microviscosity was found in BBM from RCC. Therefore, we conclude that alteration in membrane lipid composition and fluidity may play a substantial role in reduced activity of ALP in RCC.

Sharma U; Singh SK; Pal D; Khajuria R; Mandal AK; Prasad R

2012-10-01

189

Calcifying human aortic smooth muscle cells express different bone alkaline phosphatase isoforms, including the novel B1x isoform.  

UK PubMed Central (United Kingdom)

BACKGROUND: Vascular calcification, causing cardiovascular morbidity and mortality, is associated with hyperphosphatemia in chronic kidney disease (CKD). In vitro, phosphate induces transdifferentiation of vascular smooth muscle cells to osteoblast-like cells that express alkaline phosphatase (ALP). In vivo, raised serum ALP activities are associated with increased mortality. A new bone ALP isoform (B1x) has been identified in serum from CKD patients. The present study investigated the different ALP isoforms in calcifying human aortic smooth muscle cells (HAoSMCs). METHODS: HAoSMCs were cultured for 30 days in medium containing 5 or 10 mmol/l ?-glycerophosphate in the presence or absence of the ALP-specific inhibitor tetramisole. RESULTS: All known bone-specific ALP (BALP) isoforms (B/I, B1x, B1 and B2) were identified in HAoSMCs. ?-Glycerophosphate stimulated calcification of HAoSMCs, which was associated with increased BALP isoforms B/I, B1x and B2. Tetramisole inhibited the ?-glycerophosphate-induced HAoSMC calcification, which was paralleled by the inhibition of the B1x and B/I, but not the other isoforms. CONCLUSIONS: HAoSMCs express the four known BALP isoforms. B/I, B1x and B2 could be essential for soft tissue calcification. B/I and B1x were more affected by tetramisole than the other isoforms, which suggests different biological functions during calcification of HAoSMCs.

Haarhaus M; Arnqvist HJ; Magnusson P

2013-01-01

190

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

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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.

Weiss, M.J.; Cole, D.E.C.; Ray, K.; Whyte, M.P.; Lafferty, M.A.; Mulivor, R.A.; Harris, H. (Univ. of Pennsylvania School of Medicine, Philadelphia (USA))

1988-10-01

191

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

192

Mercaptopyruvate inhibits tissue-nonspecific alkaline phosphatase and calcium pyrophosphate dihydrate crystal dissolution.  

UK PubMed Central (United Kingdom)

OBJECTIVE: The enzymatic activities of tissue-nonspecific alkaline phosphatase (TNAP) including capacity to inhibit calcium pyrophosphate dihydrate (CPPD) crystal dissolution are known to be inhibited by endogenous amino acids, notably cysteine. As cysteine is recognized as a strong TNAP inhibitor, we investigated whether cysteine-related metabolites such as mercaptopyruvate (MPA) could show similar enzyme inhibition effects and, if so, whether these effects might be synergistic with cysteine at approximate physiologic concentrations of the amino acids. METHODS: We studied the inhibitory effects of MPA as well as MPA and cysteine combined in equimolar concentrations on TNAP's phosphatase, inorganic pyrophosphatase, and CPPD crystal dissolution activities. Kinetic parameters V(max), K(M), concentration for 50% inhibition (I(50)), inhibitor constant (K(I)), and specific activities calculated from initial velocity, Eadie-Hofstee, Simple, Dixon, and secondary plots were used to assess enzyme inhibition. RESULTS: MPA significantly inhibited TNAP's phosphatase and pyrophosphatase activities at 10x and 100x physiological concentrations. In the presence of calcium [Ca(2+)] and [Mg(2+)] = 1 mM, MPA inhibited uncompetitively TNAP's phosphatase activity and inhibited noncompetitively its pyrophosphatase activity. CPPD crystal dissolution activity was also inhibited. Cysteine and MPA together in equimolar concentrations inhibited TNAP enzyme activities and CPPD crystal dissolution much more effectively than MPA or cysteine alone, reducing CPPD dissolution to 38% of controls at approximate physiologic inhibitor concentrations. CONCLUSION: Endogenous amino acids like cysteine and its derivative MPA have the capacity to inhibit TNAP activities at physiologic concentrations. Downregulation of their inhibiting concentration in the cartilage interstitial fluid environment may provide a therapeutic avenue to controlled dissolution of CPPD crystal deposition in tissues.

Kannampuzha JV; Tupy JH; Pritzker KP

2009-12-01

193

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

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Full Text Available 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 (PAGE). The obtained results demonstrate a complete homology of the DNA sequence between the cloned alkaline phosphatase gene with the sequence present in the gene banks.

Hamid Mir Mohammad Sadeghi; Bahram Haghighi; Mohsen Bagheri

2005-01-01

194

Modulation of symbiont lipid A signaling by host alkaline phosphatases in the squid-vibrio symbiosis.  

UK PubMed Central (United Kingdom)

UNLABELLED: The synergistic activity of Vibrio fischeri lipid A and the peptidoglycan monomer (tracheal cytotoxin [TCT]) induces apoptosis in the superficial cells of the juvenile Euprymna scolopes light organ during the onset of the squid-vibrio symbiosis. Once the association is established in the epithelium-lined crypts of the light organ, the host degrades the symbiont's constitutively produced TCT by the amidase activity of a peptidoglycan recognition protein (E. scolopes peptidoglycan recognition protein 2 [EsPGRP2]). In the present study, we explored the role of alkaline phosphatases in transforming the lipid A of the symbiont into a form that changes its signaling properties to host tissues. We obtained full-length open reading frames for two E. scolopes alkaline phosphatase (EsAP) mRNAs (esap1 and esap2); transcript levels suggested that the dominant light organ isoform is EsAP1. Levels of total EsAP activity increased with symbiosis, but only after the lipid A-dependent morphogenetic induction at 12 h, and were regulated over the day-night cycle. Inhibition of total EsAP activity impaired normal colonization and persistence by the symbiont. EsAP activity localized to the internal regions of the symbiotic juvenile light organ, including the lumina of the crypt spaces where the symbiont resides. These data provide evidence that EsAPs work in concert with EsPGRPs to change the signaling properties of bacterial products and thereby promote persistent colonization by the mutualistic symbiont. IMPORTANCE: The potential for microbe-associated molecular patterns (MAMPs) to compromise host-tissue health is reflected in the often-used nomenclature for these molecules: lipopolysaccharide (LPS) is also called "endotoxin" and the peptidoglycan monomer is also called "tracheal cytotoxin" (TCT). With constant presentation of MAMPs by the normal microbiota, mechanisms to tolerate their effects have developed. The results of this contribution provide evidence that host alkaline phosphatases (APs) dephosphorylate and inactivate the symbiont MAMP lipid A. As such, APs work in synergy with a peptidoglycan recognition protein, which inactivates symbiont-exported TCT, to alter the symbiont MAMPs and promote persistence of the partnership. Not only may these activities serve to "tame" the MAMPs, but also the resulting products may themselves be important signals in persistent mutualisms. The finding of lipid A modification by APs in an invertebrate mutualism provides evidence that this specific strategy for dealing with symbiotic partners is conserved across the animal kingdom.

Rader BA; Kremer N; Apicella MA; Goldman WE; McFall-Ngai MJ

2012-01-01

195

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

196

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

International Nuclear Information System (INIS)

[en] 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 Mg2+.

2012-01-01

197

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

Science.gov (United States)

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

Grunwald, Sandra K.; Krueger, Katherine J.

2008-01-01

198

Enzymatic mineralization of hydrogels for bone tissue engineering by incorporation of alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase (ALP), an enzyme involved in mineralization of bone, is incorporated into three hydrogel biomaterials to induce their mineralization with calcium phosphate (CaP). These are collagen type I, a mussel-protein-inspired adhesive consisting of PEG substituted with catechol groups, cPEG, and the PEG/fumaric acid copolymer OPF. After incubation in Ca-GP solution, FTIR, EDS, SEM, XRD, SAED, ICP-OES, and von Kossa staining confirm CaP formation. The amount of mineral formed decreases in the order cPEG?>?collagen?>?OPF. The mineral:polymer ratio decreases in the order collagen?>?cPEG?>?OPF. Mineralization increases Young's modulus, most profoundly for cPEG. Such enzymatically mineralized hydrogel/CaP composites may find application as bone regeneration materials.

Douglas TE; Messersmith PB; Chasan S; Mikos AG; de Mulder EL; Dickson G; Schaubroeck D; Balcaen L; Vanhaecke F; Dubruel P; Jansen JA; Leeuwenburgh SC

2012-08-01

199

New fracture-labelling method: alkaline phosphatase in unstimulated human neutrophils.  

UK PubMed Central (United Kingdom)

We show a new freeze-fracture enzyme cytochemistry technique, i.e. fracture-labelling by enzyme cytochemistry. Freeze-fracture replication is carried out first and subsequently enzyme molecules in the split-membrane halves of cellular membranes are labelled with enzyme cytochemical markers. A replica-digestion treatment before the cytochemical reactions is a key step in this method. Triton X-100, saponin, and ultrasonication provided adequate cleaning of the replicas with good preservation of enzyme activity. Enzyme cytochemical fracture-labelling was applied to the study of alkaline phosphatase (ALPase)-positive granules in unstimulated human neutrophils. ALPase activity was found primarily on the exoplasmic membrane halves of intracellular small granules. Using X-ray microanalysis, we confirmed that the electron-dense deposit on replicas was the reaction product demonstrating ALPase activity. The results obtained by this method should provide unique information for the understanding of structure and function of biological membranes.

Takizawa T; Saito T

1997-01-01

200

Alkaline Phosphatase and CD34 Reaction of Deciduous Teeth Pulp Stem Cells  

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Full Text Available Endothelial progenitor cells from the pulp of milk teeth were isolated for use in clinical applications and tissue engineering. Normal deciduous teeth from children of 7 to 8 years of age, which more than half the tooth root was extracted, were selected from the dental centre. Cells from enzyme treated pulps were cultured and cells resulting from the fifth and eight subculture were combined for cell surface marker determination experiments. Cells were positive for CD34 marker with a total of 99/45%, determined by flowcytometry. Cells also demonstrated alkaline phosphatase (ALP) activity. From the developmental point of view, stem cells from the dental pulp seem to have derived from the neural crest, which our findings technically support this theory. In essence mobile progenitor cells from bone marrow of endothelial origin could also play a significant role in the derivation of dental pulp stem cells.

Fatemeh Abedini; Tahereh Foroutan; Leila Jahangiri

2007-01-01

 
 
 
 
201

Fluoride stimulates ( sup 3 H)thymidine incorporation and alkaline phosphatase production by human osteoblasts  

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The effect of sodium fluoride on alkaline phosphatase (ALP) release and ({sup 3}H)thymidine uptake by human osteoblasts in culture was investigated. Sodium fluoride stimulated both ALP release and ({sup 3}H)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.

Khokher, M.A.; Dandona, P. (Royal Free Hospital, London (England))

1990-11-01

202

Bezafibrate normalizes alkaline phosphatase in primary biliary cirrhosis patients with incomplete response to ursodeoxycholic acid.  

UK PubMed Central (United Kingdom)

BACKGROUND & AIMS: Ursodeoxycholic acid (UDCA) is the standard treatment for primary biliary cirrhosis (PBC) but excellent response is not observed in all cases. Since potential favourable effects of fibrates have been reported in short series with inconclusive results, we have carried out a pilot study to analyse the effects of bezafibrate in patients with suboptimal response to UDCA. METHODS: Thirty women (age 52.3 ± 2.3 years) treated with UDCA and abnormal alkaline phosphatase (AP) levels received bezafibrate (400 mg/d) for 1 year. Changes were measured every 3 months during the study period of 12 months, 3 months after discontinuation and 3 months after resuming bezafibrate. RESULTS: Two patients discontinued the treatment after few days, three at 6 and one at 9 months. Bezafibrate treatment resulted in a significant decrease in AP as early as 3 months. Normalization or decrease of AP below 1.5 times normal levels was observed in 13 and 4 patients respectively. There was also a significant decrease in ?-glutamyl transferase and alanine aminotransferase, cholesterol and triglyceride levels. Bezafibrate treatment resulted in significant improvement of pruritus. A rebound in liver biochemistries and pruritus occurred upon drug discontinuation, changes which improved again after resuming bezafibrate. Response to bezafibrate was associated with lower liver stiffness and severity of cholestasis. No severe adverse effects were observed. CONCLUSIONS: Combination treatment of bezafibrate and UDCA is associated with marked decrease or normalization of alkaline phosphatase as early as 3 months in patients with PBC. Better biochemical response was observed in patients with early disease and lower cholestasis.

Lens S; Leoz M; Nazal L; Bruguera M; Parés A

2013-07-01

203

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

UK PubMed Central (United Kingdom)

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.

Lin HY; Shih CY; Liu HC; Chang J; Chen YL; Chen YR; Lin HT; Chang YY; Hsu CH; Lin HJ

2013-08-01

204

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

UK PubMed Central (United Kingdom)

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.

Mazzei P; Oschkinat H; Piccolo A

2013-08-01

205

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

UK PubMed Central (United Kingdom)

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.

Chung TD

2013-01-01

206

Immobilization of alkaline phosphatase on magnetic particles by site-specific and covalent cross-linking catalyzed by microbial transglutaminase.  

UK PubMed Central (United Kingdom)

Bacterial alkaline phosphatase (BAP) was site-specifically and covalently immobilized on magnetic particles (MPs) using the enzymatic reaction of microbial transglutaminase (MTG). Immobilization efficiency was affected by the chemical surface treatment of MPs and immobilized BAP exhibited more than 90% of the initial activity after 10 rounds of recycling.

Moriyama K; Sung K; Goto M; Kamiya N

2011-06-01

207

Concentrating Autographa californica nuclear polyhedrosis virus and recombinant alkaline phosphatase from insect cells using a temperature-sensitive hydrogel  

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Recombinant alkaline phosphatase expressed in insect cells was concentrated by a factor of one and half times at a separation efficiency of 54.2% using hydrogel ultrafiltration. Enzyme concentration was confirmed by SDS-PAGE as well as by spectrophotometric measurement. Wild and recombinant Autograp...

Park, Jong Hwa; Park, Chang-Ho; Chung, In Sik

208

Dissolved Phosphorus Pools and Alkaline Phosphatase Activity in the Euphotic Zone of the Western North Pacific Ocean  

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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 f...

Suzumura, Masahiro; Hashihama, Fuminori; Yamada, Namiha; Kinouchi, Shinko

209

Lactoperoxidase-125I localization of salt-extractable alkaline phosphatase on the cytoplasmic membrane of Bacillus licheniformis.  

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Previous histochemical and biochemical localizations of alkaline phosphatase in Bacillus licheniformis MC14 have shown that the membrane-associated form of the enzyme is located on the inner surface of the cytoplasmic membrane, and soluble forms are located in the periplasmic space and in the growth...

Spencer, D B; Hulett, F M

210

Relation of neutrophil alkaline phosphatase activity to Fc IgG receptor development in human blood and bone marrow.  

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A possible correlation between Fc-IgG receptor expression and neutrophil alkaline phosphatase (NAP) activity was investigated in relation to maturation of granulocytes in human peripheral blood and bone marrow. NAP activity was studied in bone marrow from patients with normal peripheral blood NAP sc...

Scott, C S; Cordingley, R J; Roberts, B E; Bynoe, A G; Hough, D

211

Tissue-nonspecific alkaline phosphatase and plasma cell membrane glycoprotein-1 are central antagonistic regulators of bone mineralization  

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Osteoblasts mineralize bone matrix by promoting hydroxyapatite crystal formation and growth in the interior of membrane-limited matrix vesicles (MVs) and by propagating the crystals onto the collagenous extracellular matrix. Two osteoblast proteins, tissue-nonspecific alkaline phosphatase (TNAP) and...

Hessle, Lovisa; Johnson, Kristen A.; Anderson, H. Clarke; Narisawa, Sonoko; Sali, Adnan; Goding, James W.; Terkeltaub, Robert

212

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

213

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

Science.gov (United States)

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

Kubo, Keitaro; Yuki, Kazuhito; Ikebukuro, Toshihiro

2012-01-01

214

The effect of 50 kV X-ray irradiation on the alkaline phosphatase activity of growing rat bone  

International Nuclear Information System (INIS)

Alkaline phosphatase activity was decreased in tibial metaphysis of growing rats on the first day after 50 kV x-irradiation with 0.5-8.0 Gy. There were no differences in enzyme activity between the control and the irradiated metaphysis 30 days after irradiation. (author)

1981-01-01

215

Use of monoclonal antibodies to assign phenotypes to placental alkaline phosphatase from cultured human cell lines derived from tumors.  

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Monoclonal antibodies were used to type placental alkaline phosphatase (ALP) from cell lines established from malignant human tumors by incubating ALP extracts from the cells with antibodies of different allelic specificities and separating free from bound enzyme on polyacrylamide gel electrophoresi...

Wray, L K; Harris, H

216

Abnormal alkaline phosphatase of hepatic type in cerebrospinal fluid of a patient with intracranial metastasis from lung cancer.  

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High alkaline phosphatase (ALP) activity was found in the cerebrospinal fluid of a patient with intracranial metastases from adenocarcinoma of the lung. On agarose gel electrophoresis of the major ALP isoenzyme found in the cerebrospinal fluid, its mobility was different from those of the usual seru...

Hoshino, T; Kumasaka, K; Kawano, K; Koyama, I; Arai-Fujimori, Y; Yamagishi, F; Sakagishi, Y; Komoda, T

217

Chemiluminescent assay of alkaline phosphatase using dihydroxyacetone phosphate as substrate detected with lucigenin.  

Science.gov (United States)

A sensitive and simple chemiluminescent assay (CL) for alkaline phosphatase (ALP) using dihydroxyacetone phosphate or its ketal (DHAP or DHAP-ketal) was developed. New substrates were transformed to dihydroxyacetone (DHA) after they were hydrolysed by ALP, which reacts with lucigenin and produces strong chemiluminescence. Under the optimum assay condition, the detection limits were 3.8 x 10(-19) and 1.5 x 10(-18) moles of ALP, respectively. The coefficients of variation (CV) at each points on the standard curve were 0.8-5.4% and 1.8-7.1% (n = 6), respectively. The mechanism of lucigenin CL with DHA was investigated by ESR spectrometry using the spin-trapping method. The mechanism was speculated as follows: the O2(-) generated by the reaction of DHA and O2 in alkaline solution reacts with lucigenin, and then emit light. The proposed CL assay was applied to the enzyme immunoassay of 17beta-oestradiol, using ALP as a label enzyme. The measurable range of 17beta-oestradiol was 15-4000-pg/mL, and the proposed method was four times more sensitive than the colorimetric assay for ALP by using 4-nitrophenyl phosphate as substrate. PMID:11816056

Kokado, Amane; Arakawa, Hidetoshi; Maeda, Masako

218

Chemiluminescent assay of alkaline phosphatase using dihydroxyacetone phosphate as substrate detected with lucigenin.  

UK PubMed Central (United Kingdom)

A sensitive and simple chemiluminescent assay (CL) for alkaline phosphatase (ALP) using dihydroxyacetone phosphate or its ketal (DHAP or DHAP-ketal) was developed. New substrates were transformed to dihydroxyacetone (DHA) after they were hydrolysed by ALP, which reacts with lucigenin and produces strong chemiluminescence. Under the optimum assay condition, the detection limits were 3.8 x 10(-19) and 1.5 x 10(-18) moles of ALP, respectively. The coefficients of variation (CV) at each points on the standard curve were 0.8-5.4% and 1.8-7.1% (n = 6), respectively. The mechanism of lucigenin CL with DHA was investigated by ESR spectrometry using the spin-trapping method. The mechanism was speculated as follows: the O2(-) generated by the reaction of DHA and O2 in alkaline solution reacts with lucigenin, and then emit light. The proposed CL assay was applied to the enzyme immunoassay of 17beta-oestradiol, using ALP as a label enzyme. The measurable range of 17beta-oestradiol was 15-4000-pg/mL, and the proposed method was four times more sensitive than the colorimetric assay for ALP by using 4-nitrophenyl phosphate as substrate.

Kokado A; Arakawa H; Maeda M

2002-01-01

219

Hyperbilirubinemia in the early phase after allogeneic HSCT: prognostic significance of the alkaline phosphatase/total bilirubin ratio.  

UK PubMed Central (United Kingdom)

Hyperbilirubinemia in the early phase after allogeneic hematopoietic SCT (HSCT) is due to various causes. One of the most important causes of hyperbilirubinemia is veno-occlusive disease/sinusoidal obstructive syndrome (VOD/SOS). However, the prognosis of patients who are clinically diagnosed as SOS varies. We retrospectively evaluated 82 patients who underwent their first allogeneic HSCT. GVHD prophylaxis was a combination of short-term MTX and CsA (n=77) or tacrolimus (n=5). Thirty-three patients developed hyperbilirubinemia, with a bilirubin level of at least 2?mg/dL, within 20 days after HSCT. Of these patients, 24 were diagnosed as VOD/SOS using the modified Seattle criteria. Twenty-six recovered to a bilirubin level of <2?mg/dL. We focused on the serum alkaline phosphatase/total bilirubin ratio (ALP/TB) at the onset of hyperbilirubinemia and found that it significantly predicted the recovery from hyperbilirubinemia. OS was significantly higher in patients with a lower ALP/TB ratio (P=0.00056). In addition, a lower ALP/TB ratio was associated with better survival even in patients who were clinically diagnosed as SOS (P<0.001). The ALP/TB ratio at the onset of hyperbilirubinemia may be a useful predictor for the prognosis of hyperbilirubinemia and SOS early after HSCT.

Ashizawa M; Oshima K; Wada H; Ishihara Y; Kawamura K; Sakamoto K; Sato M; Terasako K; Machishima T; Kimura S; Kikuchi M; Nakasone H; Okuda S; Kako S; Kanda J; Yamazaki R; Tanihara A; Nishida J; Kanda Y

2013-01-01

220

Fluorescent Light-up Probe with Aggregation-Induced Emission Characteristics for Alkaline Phosphatase Sensing and Activity Study.  

UK PubMed Central (United Kingdom)

Fluorogens with aggregation-induced emission (AIE) characteristics have attracted intensified research interest in biosensing applications, and those with specific targeting ability are especially desirable. In this work, we designed and synthesized an AIE fluorescent probe by functionalizing a tetraphenylethylene (TPE) fluorogen with two phosphate groups (TPE-phos) for the detection of alkaline phosphatase (ALP) and its enzymatic activity based on the specific interaction between the probe and ALP. The probe is virtually non-fluorescent in aqueous media due to good water solubility. In the presence of ALP, the phosphate groups are cleaved through enzymatic hydrolysis, yielding a highly fluorescent product as a result of activated AIE process. This light-up probe shows excellent selectivity towards ALP among a group of proteins. The detection limit is found to be 11.4 pM or 0.2 U L-1 in Tris buffer solution with a linear quantification range of 3-526 U L-1. The assay is also successfully performed in diluted serum with a linear range up to 175 U L-1, demonstrating its potential application in clinical analysis of ALP levels in real samples. Furthermore, by conducting kinetic analysis of the enzyme using TPE-phos as the substrate, the kinetic parameter kcat/KM is determined to be 4.0 × 105 M-1 s-1, indicating a high efficiency of the substrate.

Liang J; Kwok RT; Shi H; Tang BZ; Liu B

2013-08-01

 
 
 
 
221

Alkaline phosphatase and its isoenzyme activity for the evaluation of bone metabolism in children receiving anticonvulsant monotherapy.  

Science.gov (United States)

This study aimed to investigate whether carbamazepine, sodium valproate or phenobarbital as monotherapy in ambulatory epileptic children with adequate sun exposure have some effect on their bone metabolism based on the determination of total serum alkaline phosphatase (AP) levels and its bone isoenzyme activity. Blood samples were obtained from 118 epileptic children (37 on carbamazepine, 47 on sodium valproate and 34 on phenobarbital) and from corresponding healthy controls matched for age, gender and anthropometric parameters. AP and its liver, bone and intestinal isoenzyme levels, other common biochemical markers of bone and liver metabolism and drug levels were measured in the study participants. Patients on carbamazepine or phenobarbital had significantly elevated AP levels accompanied by increased bone and liver isoenzyme activity compared to controls. An increase of bone AP isoenzyme values, correlated with the duration of treatment ( r= 0.49, P= 0.002), was found in children on sodium valproate without, however, a concomitant significant elevation of total AP values. We conclude that children who receive antiepileptic drugs as monotherapy, even when residing in a Mediterranean country with adequate sunlight, may have their bone metabolism affected as indicated by the elevated levels of bone AP isoenzyme. This isoenzyme, but not total AP values, could therefore be used as a marker for the selection of patients who would be benefited by a thorough evaluation of their bone metabolism profile. PMID:12160665

Voudris, Konstantinos; Moustaki, Maria; Zeis, Petros M; Dimou, Stamatia; Vagiakou, Eleni; Tsagris, Basilios; Skardoutsou, Angeliki

2002-09-01

222

Fluorescent Light-up Probe with Aggregation-Induced Emission Characteristics for Alkaline Phosphatase Sensing and Activity Study.  

Science.gov (United States)

Fluorogens with aggregation-induced emission (AIE) characteristics have attracted intensified research interest in biosensing applications, and those with specific targeting ability are especially desirable. In this work, we designed and synthesized an AIE fluorescent probe by functionalizing a tetraphenylethylene (TPE) fluorogen with two phosphate groups (TPE-phos) for the detection of alkaline phosphatase (ALP) and its enzymatic activity based on the specific interaction between the probe and ALP. The probe is virtually nonfluorescent in aqueous media due to good water solubility. In the presence of ALP, the phosphate groups are cleaved through enzymatic hydrolysis, yielding a highly fluorescent product as a result of activated AIE process. This light-up probe shows excellent selectivity toward ALP among a group of proteins. The detection limit is found to be 11.4 pM or 0.2 U L(-1) in Tris buffer solution with a linear quantification range of 3-526 U L(-1). The assay is also successfully performed in diluted serum with a linear range up to 175 U L(-1), demonstrating its potential application in clinical analysis of ALP levels in real samples. Furthermore, by conducting kinetic analysis of the enzyme using TPE-phos as the substrate, the kinetic parameter kcat/KM is determined to be 5.1 × 10(5) M(-1) s(-1), indicating a high efficiency of the substrate. PMID:23957823

Liang, Jing; Kwok, Ryan Tsz Kin; Shi, Haibin; Tang, Ben Zhong; Liu, Bin

2013-08-30

223

Extracellular alkaline phosphatase is a sensitive marker for cellular stimulation and exocytosis in heterotroph cell cultures of Chenopodium rubrum.  

Science.gov (United States)

We investigated the response of extracellular phosphatase to heat shock in heterotrophic Chenopodium rubrum L. cell cultures. Surprisingly, in contrast to the generally used acid phosphatase, an extracellular alkaline phosphatase showed the most sensitive response. This phosphatase was characterized as a marker for cellular stimulation by its high correlations with induced changes of extracellular pH: 10microM nigericin (correlation coefficient r=0.91), 100microM salicylic acid (r=0.84), heat shock 5min 37 degrees C (r=0.79), and heat shock after pre-treatment with 5microM fusicoccin (r=0.92) or 0.5% ethanol (r=0.90). Cellular stimulation was estimated with concentrations of acids and bases, yielding similar levels of pH change (0.5 pH) in cell-free supernatant: salicylic acid (200microM), benzoic acid (600microM), HCl (140microM), NaOH (100microM), and KOH (100microM). The Golgi apparatus inhibitor Brefeldin A (200microM) reduced the heat-shock-induced phosphatase (-33%). The pH optimum of heat-shock-induced phosphatase was 3; however, there the proportion of constitutive phosphatase was higher than at pH 8-9.5, indicating different pH dependence of constitutive and induced activity. Thus, heat-shock-induced phosphatase was characterized by alkaline activity with inhibitors (10microM molybdate: -52%, 2.5mM phosphate: -64%, 10microM ZnCl(2): -82%), substrates (2.5mM, tyrosine phosphate: 255pkat g(-1), p-nitrophenyl phosphate: 92pkat g(-1), serine phosphate: 0, threonine phosphate: 0), Hill coefficient (nH=1.4) indicating two binding sites, and the extent of heat-shock stimulation (p-nitrophenyl phosphate: +190%, tyrosine phosphate: +180%). SDS-PAGE showed a correlation of alkaline phosphatase with the heat-shock-induced release of highly N-glycosylated 53kDa protein, detected by peroxidase-labeled concanavalin A affinoblotting after endoglycosidase H treatment. The 53kDa protein showed no in-gel phosphatase activity after SDS-PAGE and regeneration treatment, in contrast to a putative dimer (105kDa). PMID:18433930

Chaidee, Anchalee; Wongchai, Chatchawal; Pfeiffer, Wolfgang

2008-04-22

224

Activity of soil dehydrogenases, urease, and acid and alkaline phosphatases in soil polluted with petroleum.  

UK PubMed Central (United Kingdom)

This study was undertaken to (1) determine the effects of petroleum pollution on changes in the biochemical properties of soil and (2) demonstrate whether the application of compost, bentonite, and calcium oxide is likely to restore biological balance. Petroleum soil pollution at a dose ranging from 2.5 to 10 cm(3)/kg disturbed the biochemical balance as evidenced by inhibition of the activities of soil dehydrogenases (SDH), urease (URE), and acid phosphatase (ACP). The greatest change was noted in the activity of SDH, whereas the least change occurred in URE. Petroleum significantly increased the activity of soil alkaline phosphatase (ALP) in soil used for spring rape, whereas in soil used for oat harvest there was decreased ALP activity. The application of compost, bentonite, and calcium oxide to soil proved effective in mitigating the adverse effects of petroleum on the activities of soil enzymes. Soil enrichment with compost, bentonite, and calcium oxide was found to stimulate the activities of URE and ALP and inhibit the activity of ACP. The influence of bentonite and calcium oxide was greater than that of compost. Calcium oxide and, to a lesser extent, compost were found to increase the activity of SDH, whereas bentonite exerted the opposite effect, especially in the case of the main crop, spring rape. The activities of SDH, URE, and ACP were higher in soil used for rape than that for oats. In contrast the activity of ALP was higher in soil used for oats. Data thus indicate that compost and especially bentonite and calcium oxide exerted a positive effect on activities of some enzymes in soil polluted with petroleum. Application of neutralizing additives to soil restored soil biological balance by counteracting the negative influence of petroleum on activities of URE and ALP.

Wyszkowska J; Wyszkowski M

2010-01-01

225

ALKALINE PHOSPHATASE CmAP SYNTHESIS-DETERMINING 40Ph PLASMID, E. coli rosetta(DE3)/40Ph STRAIN - PRODUCER OF CHIMERIC PROTEIN, CONTAINING AMINO ACID SEQUENCE OF RECOMBINANT ALKALINE PHOSPHATASE CmAP, AND PRODUCTION METHOD THEREOF  

UK PubMed Central (United Kingdom)

FIELD: chemistry. ^ SUBSTANCE: disclosed is a plasmid which determines synthesis of alkaline phosphatase CmAP, containing a NcoI/SacI-fragment of plasmid pET-40b(+) (Novagen) and a DNA fragment of 1530 base pairs, which contains a chimeric gene consisting of a CmAP gene structural part which is adapted at the N-end for expression in E.coli cells, and a nucleotide which codes a specific sequence for TEV protease. Described is an E.coli Rosetta(DE3) strain which is transformed by said plasmid, and is a producer of chimeric protein, which contains an amino acid sequence of recombinant alkaline phosphatase CmAP. Disclosed is a method of obtaining recombinant alkaline phosphatase CmAP, comprising steps for: incubating said producer strain in a liquid broth LB for 12 hours at 20C, depositing bacterial cells by centrifuging, disintegration of the cell suspension in a buffer, centrifuging the extract, chromatography of the supernatant fluid on a column with a metal affinity resin, elution of the protein, concentrating active fractions by ultrafiltration, incubation with protease TEV, concentrating the protein solution and extracting the end product by gel filtration. ^ EFFECT: improved method. ^ 3 cl, 1 dwg, 3 ex

BALABANOVA LARISA ANATOL EVNA; RASSKAZOV VALERIJ ALEKSANDROVICH

226

Gingival crevicular fluid alkaline phosphatase activity as a non-invasive biomarker of skeletal maturation.  

UK PubMed Central (United Kingdom)

OBJECTIVES: To evaluate the gingival crevicular fluid (GCF) alkaline phosphatase (ALP) activity in growing subjects in relation to the stages of individual skeletal maturation. SETTING AND SAMPLE POPULATION: The Department of Biomedicine, University of Trieste. Seventy-two healthy growing subjects (45 women and 27 men; range, 7.8-17.7?years). MATERIALS AND METHODS: Double-blind, prospective, cross-sectional design. Samples of GCF were collected from each subject at the mesial and distal sites of both of the central incisors, in the maxilla and mandible. Skeletal maturation phase was assessed through the cervical vertebral maturation (CVM) method. Enzymatic activity was determined spectrophotometrically. RESULTS: The relationship between GCF ALP activity and CVM stages was significant. In particular, a twofold peak in enzyme activity was seen at the CS3 and CS4 pubertal stages, compared to the pre-pubertal stages (CS1 and CS2) and post-pubertal stages (CS5 and CS6), at both the maxillary and mandibular sites. No differences were seen between the maxillary and mandibular sites, or between the sexes. CONCLUSIONS: As an adjunct to standard methods based upon radiographic parameters, the GCF ALP may be a candidate as a non-invasive clinical biomarker for the identification of the pubertal growth spurt in periodontally healthy subjects scheduled for orthodontic treatment.

Perinetti G; Baccetti T; Contardo L; Di Lenarda R

2011-02-01

227

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

Directory of Open Access Journals (Sweden)

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

2011-01-01

228

Structural study on the carbohydrate moiety of human placental alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase purified from human placenta contains a single asparagine-linked sugar chain in one molecule. The sugar chain was quantitatively liberated as radioactive oligosaccharides from the polypeptide moiety by hydrazinolysis followed by N-acetylation and NaB3H4 reduction, and separated by paper electrophoresis into one neutral and two acidic fractions. By a combination of sequential exoglycosidase digestion and methylation analysis, the structures of oligosaccharides in the neutral fraction were confirmed to be as follows: Gal beta 1----4GlcNAc beta 1----2Man alpha 1----6(Gal beta 1----4GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4(+/- Fuc alpha 1----6)GlcNAc. The acidic oligosaccharide fractions were mixtures of mono- and disialyl derivatives of the neutral fraction. All the sialic acid residues of the sugar chains occur as the NeuAc alpha 2----3Gal group. In the case of monosialyl derivatives, the N-acetylneuraminic acid was exclusively linked to the Man alpha 1----3 arm.

Endo T; Ohbayashi H; Hayashi Y; Ikehara Y; Kochibe N; Kobata A

1988-01-01

229

Structural study on the carbohydrate moiety of human placental alkaline phosphatase.  

Science.gov (United States)

Alkaline phosphatase purified from human placenta contains a single asparagine-linked sugar chain in one molecule. The sugar chain was quantitatively liberated as radioactive oligosaccharides from the polypeptide moiety by hydrazinolysis followed by N-acetylation and NaB3H4 reduction, and separated by paper electrophoresis into one neutral and two acidic fractions. By a combination of sequential exoglycosidase digestion and methylation analysis, the structures of oligosaccharides in the neutral fraction were confirmed to be as follows: Gal beta 1----4GlcNAc beta 1----2Man alpha 1----6(Gal beta 1----4GlcNAc beta 1----2Man alpha 1----3)Man beta 1----4GlcNAc beta 1----4(+/- Fuc alpha 1----6)GlcNAc. The acidic oligosaccharide fractions were mixtures of mono- and disialyl derivatives of the neutral fraction. All the sialic acid residues of the sugar chains occur as the NeuAc alpha 2----3Gal group. In the case of monosialyl derivatives, the N-acetylneuraminic acid was exclusively linked to the Man alpha 1----3 arm. PMID:3360758

Endo, T; Ohbayashi, H; Hayashi, Y; Ikehara, Y; Kochibe, N; Kobata, A

1988-01-01

230

Unfolding and inactivation of abalone (Haliotis diversicolor) alkaline phosphatase during denaturation by guanidine hydrochloride.  

UK PubMed Central (United Kingdom)

Abalone, a kind of low poikilothermic invertebrate, is easily exposed to ocean environment stress. Since it is one of the important mariculture animals, the attention paid to the abalone study becomes increasing. Alkaline phosphatase (ALPase, EC 3.1.3.1) is a kind of zinc-contained metalloenzyme, which catalyzes the nonspecific hydrolysis of phosphate monoesters. Unfolding and inactivation of ALPase from abalone (Haliotis diversicolor) during denaturation by guanidine hydrochloride (GuHCl) of different concentrations has first been studied. The kinetic theory of the substrate reaction by enzyme was described by Tsou, which was applied to the study on ALPase's kinetic course of inactivation by GuHCl. The result showed that the inactivation of the enzyme by GuHCl was a slow, reversible reaction with fractional remaining activity. The microscopic rate constants were determined. The result, k(+0) > k'(+0), showed that the enzyme was protected by the substrate to a certain extent during guanidine denaturation. The changes of conformation of the enzyme in different concentrations of GuHCl have been studied by means of measuring the fluorescence spectra. The results showed that the inactivation occurred before the noticeable conformational changes of the enzyme molecule as a whole can be detected, which suggests that the active site of the enzyme has more flexibility than the whole enzyme molecule. These studies will facilitate the understanding of physiological and biochemical features of the H. diversicolor and will also help in the understanding of the abalone immune system.

Liao JH; Chen QX; Zhang Q; Yang Y; Shi Y

2009-08-01

231

A moderately thermostable alkaline phosphatase from Geobacillus thermodenitrificans T2: cloning, expression and biochemical characterization.  

Science.gov (United States)

A gene-encoding alkaline phosphatase (AP) from thermophilic Geobacillus thermodenitrificans T2, termed Gtd AP, was cloned and sequenced. The deduced Gtd AP protein comprises 424 amino acids and shares a low homology with other known AP (Gtd AP protein, without a predicted signal peptide of 30 amino acids, was successfully overexpressed in E. coli and purified as a hexa-His-tagged fusion protein. The pH and temperature optima for purified enzyme are 9.0 and 65 degrees C, respectively. The enzyme retained a high activity at 45-60 degrees C, while it could be quickly inactivated by a heat treatment at 80 degrees C for 15 min, exhibiting a half-life of 8 min at 70 degrees C. The K(m) and V (max) for pNPP were determined to be 31.5 muM and 430 muM/min at optimal conditions. A divalent cation is essential, with a combination of Mg2+ and Co2+ or Zn2+ preferred. The enzyme was strongly inhibited by 10 mM ethylenediaminetetraacetic acid (EDTA) and vanadate but highly resistant to urea and dithiothreitol. The properties of Gtd AP make it suitable for application in molecular cloning or amplification. PMID:18365148

Zhang, Yong; Ji, Chaoneng; Zhang, Xiaoxiao; Yang, Zhenxing; Peng, Jing; Qiu, Rui; Xie, Yi; Mao, Yumin

2008-03-26

232

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

Directory of Open Access Journals (Sweden)

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; Crisciele Kuligovski; Marise Teresinha Brenner Affonso da Costa; Marco Augusto Stimamiglio; Carmen Lúcia Kuniyoshi Rebelatto; Alexandra Cristina Senegaglia; Paulo Roberto Slud Brofman; Bruno Dallagiovanna; Samuel Goldenberg; Alejandro Correa

2011-01-01

233

Bacillus subtilis transcription regulator, Spo0A, decreases alkaline phosphatase levels induced by phosphate starvation.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase (APase) is induced as a culture enters stationary phase because of limiting phosphate. The results presented here show that expression of APase is regulated both negatively and positively. PhoP, a homolog of a family of bacterial transcription factors, and PhoR, a homolog of bacterial histidine protein kinases, are required for induction of APases when phosphate becomes limiting. The induction period lasts 2 to 3 h, after which the rate of APase accumulation is decreased. Mutant strains defective in the Spo0A transcription factor failed to decrease APase production. The consequent hyperinduction of APase in a spo0A strain was dependent on phoP and phoR. spo0B and spo0F strains also overexpressed APase, suggesting that phosphorylated Spo0A is required for repression of APase. An abrB mutant allele in the presence of the mutant spo0A allele in these strains did not significantly change the APase hyperinduction phenotype, demonstrating that Spo0A repression of abrB expression is not the mechanism by which Spo0A-P regulates APase expression. Our previous report that spo0A mutants do not express APases is in conflict with the present data. We show here that the previously used mutants and a number of commonly used spo0 strains, all of which have an APase deficiency phenotype, contain a previously unrecognized mutation in phoR.

Jensen KK; Sharkova E; Duggan MF; Qi Y; Koide A; Hoch JA; Hulett FM

1993-06-01

234

Bacillus subtilis transcription regulator, Spo0A, decreases alkaline phosphatase levels induced by phosphate starvation.  

Science.gov (United States)

Alkaline phosphatase (APase) is induced as a culture enters stationary phase because of limiting phosphate. The results presented here show that expression of APase is regulated both negatively and positively. PhoP, a homolog of a family of bacterial transcription factors, and PhoR, a homolog of bacterial histidine protein kinases, are required for induction of APases when phosphate becomes limiting. The induction period lasts 2 to 3 h, after which the rate of APase accumulation is decreased. Mutant strains defective in the Spo0A transcription factor failed to decrease APase production. The consequent hyperinduction of APase in a spo0A strain was dependent on phoP and phoR. spo0B and spo0F strains also overexpressed APase, suggesting that phosphorylated Spo0A is required for repression of APase. An abrB mutant allele in the presence of the mutant spo0A allele in these strains did not significantly change the APase hyperinduction phenotype, demonstrating that Spo0A repression of abrB expression is not the mechanism by which Spo0A-P regulates APase expression. Our previous report that spo0A mutants do not express APases is in conflict with the present data. We show here that the previously used mutants and a number of commonly used spo0 strains, all of which have an APase deficiency phenotype, contain a previously unrecognized mutation in phoR. PMID:8509330

Jensen, K K; Sharkova, E; Duggan, M F; Qi, Y; Koide, A; Hoch, J A; Hulett, F M

1993-06-01

235

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

Science.gov (United States)

The enzyme alkaline phosphatase (ALP) was recently proposed as an implant coating material in order to improve the biological performance of orthopedic and dental implants. The present study evaluated the in vivo bone response to electrosprayed coatings, consisting of ALP, calcium phosphate (CaP) or a combination thereof (composite coating: ALP+CaP) compared to non-coated controls (gritblasted and acid etched). A total of 80 implants (n=10) with a gap of 1.0mm, was implanted intramedullary and bilaterally into the femurs of 80 rats. After 1 and 4 weeks, bone response was evaluated qualitatively (histology) and quantitatively (histomorphometry). The results of this study show that all electrosprayed coatings (ALP, CaP, ALP+CaP) significantly improve osteoconduction compared to non-coated controls after 4 weeks of implantation, without significant differences among these coated groups. Consequently, the results indicate that ALP-coatings improve the osteogenic response to a comparable extent as CaP-coatings or an ALP+CaP composite coating. In conclusion, the current study proofs that ALP-coatings have potential as bone implant coatings, though long-term data remain to be obtained. From a clinical perspective, it was observed that the process of osteoconduction is related to positional determinants, which needs to be taken into account when analyzing data on bone response. PMID:19717187

Schouten, C; van den Beucken, Jeroen J J P; de Jonge, Lise T; Bronkhorst, Ewald M; Meijer, Gert J; Spauwen, Paul H M; Jansen, John A

2009-08-31

236

The effect of acute inflammation on total alkaline phosphatase activity in dogs  

Directory of Open Access Journals (Sweden)

Full Text Available The main purpose of this study was to investigate the effect of acute inflammation on total alkaline phosphatase (ALP) activity in dogs. In this study total ALP activity was determined in dogs with experimentally induced acute inflammation in order to characterize their potential value in this condition. For that, ALP concentrations were defined in plasmas from 9 mongrel male dogs (in an experimental group) and 6 mongrel male dogs (in a control group) at the age of 2 years and body weight 12-15 kg. The inflammation was reproduced by inoculation of 2 ml turpentine oil subcutaneously in lumbar region and same quantity saline in control dogs. Blood samples were collected into heparinized tubes before inoculation, then at hours 6, 24, 48, 72 and on days 7, 14, 21. The total ALP concentrations were determined with commercial kits (Human-GmbH, Germany) on an automatic biochemical analyzer (BS-3000 P, Sinnowa, LTD Nanjing China). The statistical analysis of the data was performed using one way analysis of variance (ANOVA), Statistica v.6.1 (StatSoft Inc., 2002). Statistically significant difference was not found between the groups, as well as within them. In conclusion, we can say that the total activity of ALP was not significantly affected in dogs with experimentally induced acute inflammation.

Zapryanova Dimitrinka

2013-01-01

237

Intestinal Alkaline Phosphatase Prevents Antibiotic-Induced Susceptibility to Enteric Pathogens.  

UK PubMed Central (United Kingdom)

OBJECTIVE:: To determine the efficacy of oral supplementation of the gut enzyme intestinal alkaline phosphatase (IAP) in preventing antibiotic-associated infections from Salmonella enterica serovar Typhimurium (S. Typhimurium) and Clostridium difficile. BACKGROUND:: The intestinal microbiota plays a pivotal role in human health and well-being. Antibiotics inherently cause dysbiosis, an imbalance in the number and composition of intestinal commensal bacteria, which leads to susceptibility to opportunistic bacterial infections. Previously, we have shown that IAP preserves the normal homeostasis of intestinal microbiota and that oral supplementation with calf IAP (cIAP) rapidly restores the normal gut flora. We hypothesized that oral IAP supplementation would protect against antibiotic-associated bacterial infections. METHODS:: C57BL/6 mice were treated with antibiotic(s) ± cIAP in the drinking water, followed by oral gavage of S. Typhimurium or C. difficile. Mice were observed for clinical conditions and mortality. After a defined period of time, mice were killed and investigated for hematological, inflammatory, and histological changes. RESULTS:: We observed that oral supplementation with cIAP during antibiotic treatment protects mice from infections with S. Typhimurium as well as with C. difficile. Animals given IAP maintained their weight, had reduced clinical severity and gut inflammation, and showed improved survival. CONCLUSION:: Oral IAP supplementation protected mice from antibiotic-associated bacterial infections. We postulate that oral IAP supplementation could represent a novel therapy to protect against antibiotic-associated diarrhea (AAD), C. difficile-associated disease (CDAD), and other enteric infections in humans.

Alam SN; Yammine H; Moaven O; Ahmed R; Moss AK; Biswas B; Muhammad N; Biswas R; Raychowdhury A; Kaliannan K; Ghosh S; Ray M; Hamarneh SR; Barua S; Malo NS; Bhan AK; Malo MS; Hodin RA

2013-04-01

238

Effects of root canal sealers on alkaline phosphatase in human osteoblastic cells.  

UK PubMed Central (United Kingdom)

INTRODUCTION: Histologic investigations have demonstrated that root canal sealers can induce mild to severe bone resorption. Alkaline phosphatase (ALP) is a membrane-bound glycoprotein, which is one of the osteogenic differentiation markers considered to indicate the formation of new bone. The aim of this study was to investigate the effects of an epoxy resin-based sealer AH26, a zinc oxide-eugenol-based sealer Canals, and a paste sealer N2 on the expression of ALP in human osteoblastic cell line U2OS cells. METHODS: Freshly mixed materials were filled in glass rings and eluted in 10 mL of culture medium for 1 day. Subsequently, various dilutions (final dilution: 1/2, 1/4, and 1/8) of these extraction media were prepared for this study. Cytotoxicity was measured by the almar blue dye assay. Gene expression of ALP was examined by using reverse transcription-polymerase chain reaction. ALP activity was further evaluated by using substrate assay. RESULTS: The results showed that AH26, Canals, and N2 were cytotoxic to U2OS cells in a concentration-dependent manner (P < .05). The exposure of U2OS cells to AH26 and N2 resulted in the down-regulation of ALP mRNA gene expression (P < .05). ALP activity was significantly suppressed by 3 root canal sealers (P < .05). CONCLUSIONS: The inhibition of ALP expression might play an important role in the pathogenesis of root canal sealer-induced periapical bone destruction.

Huang FM; Yang SF; Chang YC

2010-07-01

239

Novel electrochemical methodology for activity estimation of alkaline phosphatase based on solubility difference.  

UK PubMed Central (United Kingdom)

We propose a novel electrochemical detection system for alkaline phosphatase (ALP) activity using the difference in water and oil solubilities between the substrate, ferrocene ethyl phosphate ester (FcEtOPO(3)(2-)), and the enzymatic product, ferroceneethanol (FcEtOH). In this system, water droplets containing ALP and FcEtOPO(3)(2-) were placed on a Pt disk microelectrode and surrounded by a mineral oil. By the ALP-catalyzed reaction, FcEtOPO(3)(2-) was converted to FcEtOH, which was then transferred to the mineral oil from the water droplets with FcEtOPO(3)(2-) remaining in the water droplets. After partitioning FcEtOH from the water droplets, FcEtOPO(3)(2-) was detected at the Pt disk microelectrode to estimate the ALP activity. Using this novel system, the ALP activity of embryoid bodies was successfully detected. We believe that the present system will be widely applicable to ALP-based bioassays.

Ino K; Kanno Y; Arai T; Inoue KY; Takahashi Y; Shiku H; Matsue T

2012-09-01

240

Alkaline phosphatase as an indicator of true ejaculation in the rhinoceros.  

UK PubMed Central (United Kingdom)

The objective was to determine if seminal alkaline phosphatase (ALP) can serve as an indicator of true ejaculation in the rhinoceros. Concentrations of ALP activity were determined in seminal fractions collected from African black rhinos (Diceros bicornis), an African white rhino (Ceratotherium simum), and an Indian rhino (Rhinoceros unicornis) during electroejaculation. In addition, seminal fractions collected during penile massage of a Sumatran rhino (Dicerorhinus sumatrensis) were assessed. Correlations between ALP activity and sperm concentration, fraction pH, and fraction osmolality were evaluated in the Indian rhino and black rhino. Concentrations of ALP activity in rhino ejaculate fractions ranged from < 5 to 11,780 U/L and were positively correlated (P < 0.05) with sperm concentration for both Indian rhino (r = 0.995) and black rhino (r = 0.697), but did not exhibit a strong correlation with either pH or osmolality (P > 0.05). Data were insufficient for establishing meaningful correlation coefficients in the Sumatran rhino and white rhino, but preliminary results were in accordance with findings in the Indian rhino and black rhino. We concluded that ALP was present in rhinoceros semen, likely originated from the epididymides and/or testes, and could serve as a useful tool for assessing the production of ejaculatory versus pre-ejaculatory fluid in the rhinoceros.

Roth TL; Stoops MA; Robeck TR; Ball RL; Wolfe BA; Finnegan MV; O'Brien JK

2010-12-01

 
 
 
 
241

Alkaline phosphatase as an indicator of true ejaculation in the rhinoceros.  

Science.gov (United States)

The objective was to determine if seminal alkaline phosphatase (ALP) can serve as an indicator of true ejaculation in the rhinoceros. Concentrations of ALP activity were determined in seminal fractions collected from African black rhinos (Diceros bicornis), an African white rhino (Ceratotherium simum), and an Indian rhino (Rhinoceros unicornis) during electroejaculation. In addition, seminal fractions collected during penile massage of a Sumatran rhino (Dicerorhinus sumatrensis) were assessed. Correlations between ALP activity and sperm concentration, fraction pH, and fraction osmolality were evaluated in the Indian rhino and black rhino. Concentrations of ALP activity in rhino ejaculate fractions ranged from 0.05). Data were insufficient for establishing meaningful correlation coefficients in the Sumatran rhino and white rhino, but preliminary results were in accordance with findings in the Indian rhino and black rhino. We concluded that ALP was present in rhinoceros semen, likely originated from the epididymides and/or testes, and could serve as a useful tool for assessing the production of ejaculatory versus pre-ejaculatory fluid in the rhinoceros. PMID:20615535

Roth, T L; Stoops, M A; Robeck, T R; Ball, R L; Wolfe, B A; Finnegan, M V; O'Brien, J K

2010-07-07

242

Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase.  

Science.gov (United States)

Thermosensitive chitosan hydrogels containing sodium beta-glycerophosphate (?-GP), whose gelation is induced by increasing temperature to body temperature, were functionalized by incorporation of alkaline phosphatase (ALP), an enzyme involved in mineralization of bone. ALP incorporation led to acceleration of gelation upon increase of temperature for four different chitosan preparations of differing molecular weight, as demonstrated by rheometric time sweeps at 37 °C. Hydrogels containing ALP were subsequently incubated in calcium glycerophosphate (Ca-GP) solution to induce their mineralization with calcium phosphate (CaP) in order to improve their suitability as materials for bone replacement. Incorporated ALP retained its bioactivity and induced formation of CaP mineral, as confirmed by SEM, FTIR, Raman spectroscopy, XRD, ICP-OES, and increases in dry mass percentage, which rose with increasing ALP concentration and incubation time in Ca-GP solution. The results demonstrate that ALP accelerates formation of thermosensitive chitosan/?-GP hydrogels and induces their mineralization with CaP, which paves the way for applications as injectable bone replacement materials. PMID:23403025

Douglas, Timothy E L; Skwarczynska, Agata; Modrzejewska, Zofia; Balcaen, Lieve; Schaubroeck, David; Lycke, Sylvia; Vanhaecke, Frank; Vandenabeele, Peter; Dubruel, Peter; Jansen, John A; Leeuwenburgh, Sander C G

2013-02-08

243

Acceleration of gelation and promotion of mineralization of chitosan hydrogels by alkaline phosphatase.  

UK PubMed Central (United Kingdom)

Thermosensitive chitosan hydrogels containing sodium beta-glycerophosphate (?-GP), whose gelation is induced by increasing temperature to body temperature, were functionalized by incorporation of alkaline phosphatase (ALP), an enzyme involved in mineralization of bone. ALP incorporation led to acceleration of gelation upon increase of temperature for four different chitosan preparations of differing molecular weight, as demonstrated by rheometric time sweeps at 37 °C. Hydrogels containing ALP were subsequently incubated in calcium glycerophosphate (Ca-GP) solution to induce their mineralization with calcium phosphate (CaP) in order to improve their suitability as materials for bone replacement. Incorporated ALP retained its bioactivity and induced formation of CaP mineral, as confirmed by SEM, FTIR, Raman spectroscopy, XRD, ICP-OES, and increases in dry mass percentage, which rose with increasing ALP concentration and incubation time in Ca-GP solution. The results demonstrate that ALP accelerates formation of thermosensitive chitosan/?-GP hydrogels and induces their mineralization with CaP, which paves the way for applications as injectable bone replacement materials.

Douglas TE; Skwarczynska A; Modrzejewska Z; Balcaen L; Schaubroeck D; Lycke S; Vanhaecke F; Vandenabeele P; Dubruel P; Jansen JA; Leeuwenburgh SC

2013-05-01

244

Structural analysis of human adult and fetal alkaline phosphatases by cyanogen bromide peptide mapping.  

Science.gov (United States)

The adult and fetal forms of human intestinal alkaline phosphatase (ALPase; orthophosphoric-monoester phosphohydrolase, EC 3.1.3.1) are indistinguishable by a variety of analytical procedures. However, they differ electrophoretically and can be differentiated by binding studies with monoclonal antibodies. In this report, these two enzymes along with placental and liver ALPases are compared by the technique of CNBr peptide mapping, and the role of carbohydrate in generating these patterns is investigated. NaDodSO4/PAGE of CNBr digests of radiolabeled ALPases from fetal and adult intestine shows that these two isozymes share five of seven common-sized CNBr fragments. Placental ALPase shares only one common-sized fragment with either intestinal enzyme. Liver ALPase has no CNBr fragments in common with any of the others. These data indicate that fetal intestinal ALPase is not a heterodimer of one subunit each of intestinal ALPase and placental ALPase as has been postulated. CNBr digests of neuraminidase-treated enzymes reveal a change of mobility of only one CNBr band in each of fetal intestinal, placental, and liver ALPases, indicating the presence of sialic acid residues in these fragments. Periodic acid/Schiff reagent staining (specific for carbohydrate) of CNBr digests of fetal and adult intestinal ALPases reacts with only one band in each enzyme, which is the same band from the fetal enzyme shown to contain sialic acid. However, fetal and adult intestinal ALPases each contain at least one CNBr fragment of unique size that is apparently nonglycosylated. Images

Vockley, J; D'Souza, M P; Foster, C J; Harris, H

1984-01-01

245

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

Energy Technology Data Exchange (ETDEWEB)

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.

Terao, M.; Mintz, B.

1987-10-01

246

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 phosphod...

Abdulrahman M. Al-Shehri

247

Role of metal ions on the secondary and quaternary structure of alkaline phosphatase from bovine intestinal mucosa.  

Science.gov (United States)

Alkaline phosphatase (EC 3.1.3.1) from bovine intestinal mucosa (BIAP) is an homodimeric metalloenzyme, containing one Mg2+ and two Zn2+ ions in each active site. ApoBIAP, prepared using ion-chelating agents, exhibited a dramatic decrease of its hydrolase activity, concomittant to conformational changes in its quaternary structure. By rate-zonal centrifugation and electrophoresis, we demonstrated, for the first time, that the loss of divalent ions leads to some monomerization process for a metal-depleted alkaline phosphatase. Divalent ions are also involved in the secondary and tertiary structures. Metal-depletion induced more exposure of some Trp residues and hydrophobic regions to the solvent (as proved by intrinsic and ANS fluorescences). These changes might correspond to the disappearance of alpha-helices and/or turns with a concomittant appearance of unordered structures and beta-sheets (as probed by FTIR spectroscopy). For BIAP, three steps of temperature-induced changes were exhibited, while for apoBIAP, only one step was exhibited at 55 degrees C. Our work on BIAP showed two main differences with alkaline phosphatase from Escherichia coli. The loss of the divalent ions induces protein monomerization and the total recovery of enzyme activity by divalent ion addition to apoBIAP was not obtained. PMID:10584076

Bortolato, M; Besson, F; Roux, B

1999-11-01

248

Role of metal ions on the secondary and quaternary structure of alkaline phosphatase from bovine intestinal mucosa.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase (EC 3.1.3.1) from bovine intestinal mucosa (BIAP) is an homodimeric metalloenzyme, containing one Mg2+ and two Zn2+ ions in each active site. ApoBIAP, prepared using ion-chelating agents, exhibited a dramatic decrease of its hydrolase activity, concomittant to conformational changes in its quaternary structure. By rate-zonal centrifugation and electrophoresis, we demonstrated, for the first time, that the loss of divalent ions leads to some monomerization process for a metal-depleted alkaline phosphatase. Divalent ions are also involved in the secondary and tertiary structures. Metal-depletion induced more exposure of some Trp residues and hydrophobic regions to the solvent (as proved by intrinsic and ANS fluorescences). These changes might correspond to the disappearance of alpha-helices and/or turns with a concomittant appearance of unordered structures and beta-sheets (as probed by FTIR spectroscopy). For BIAP, three steps of temperature-induced changes were exhibited, while for apoBIAP, only one step was exhibited at 55 degrees C. Our work on BIAP showed two main differences with alkaline phosphatase from Escherichia coli. The loss of the divalent ions induces protein monomerization and the total recovery of enzyme activity by divalent ion addition to apoBIAP was not obtained.

Bortolato M; Besson F; Roux B

1999-11-01

249

Characterization of a Cry1Ac-receptor alkaline phosphatase in susceptible and resistant Heliothis virescens larvae.  

UK PubMed Central (United Kingdom)

We reported previously a direct correlation between reduced soybean agglutinin binding to 63- and 68-kDa midgut glycoproteins and resistance to Cry1Ac toxin from Bacillus thuringiensis in the tobacco budworm (Heliothis virescens). In the present work we describe the identification of the 68-kDa glycoprotein as a membrane-bound form of alkaline phosphatase we term HvALP. Lectin blot analysis of HvALP revealed the existence of N-linked oligosaccharides containing terminal N-acetylgalactosamine required for [125I]Cry1Ac binding in ligand blots. Based on immunoblotting and alkaline phosphatase activity detection, reduced soybean agglutinin binding to HvALP from Cry1Ac resistant larvae of the H. virescens YHD2 strain was attributable to reduced amounts of HvALP in resistant larvae. Quantification of specific alkaline phosphatase activity in brush border membrane proteins from susceptible (YDK and F1 generation from backcrosses) and YHD2 H. virescens larvae confirmed the observation of reduced HvALP levels. We propose HvALP as a Cry1Ac binding protein that is present at reduced levels in brush border membrane vesicles from YHD2 larvae.

Jurat-Fuentes JL; Adang MJ

2004-08-01

250

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 Pb(2+). 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 (I(p), 117.2) of (Rhod.S)(n)-P-SOR system, which was 2.4 times higher than that without SOR (I(p), 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 -NH(2) of alkaline phosphatase (AP) caused the RTP intensity (?I(p)) 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)gmL(-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. PMID:22858611

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

2012-07-16

251

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

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

252

Alkaline phosphatase: placental and tissue-nonspecific isoenzymes hydrolyze phosphoethanolamine, inorganic pyrophosphate, and pyridoxal 5'-phosphate. Substrate accumulation in carriers of hypophosphatasia corrects during pregnancy.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Hypophosphatasia features selective deficiency of activity of the tissue-nonspecific (liver/bone/kidney) alkaline phosphatase (ALP) isoenzyme (TNSALP); placental and intestinal ALP isoenzyme (PALP and IALP, respectively) activity is not reduced. Three phosphocompounds (phosphoethanolamine [PEA], ino...

Whyte, M P; Landt, M; Ryan, L M; Mulivor, R A; Henthorn, P S; Fedde, K N; Mahuren, J D; Coburn, S P

253

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

International Nuclear Information System (INIS)

[en] 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

254

Alkaline phosphatases contribute to uterine receptivity, implantation, decidualization and defense against bacterial endotoxin in hamsters.  

UK PubMed Central (United Kingdom)

Alkaline phosphatase (AP) activity has been demonstrated in the uterus of several species, but its importance in the uterus, in general and during pregnancy, is yet to be revealed. In this study, we focused on identifying AP isozyme types, and their hormonal regulation, cell-type and event-specific expression and possible functions in the hamster uterus during the cycle and early pregnancy. Our RT-PCR and in situ hybridization studies demonstrated that among the known Akp2, Akp3, Akp5 and Akp6 murine AP isozyme genes, hamster uteri express only Akp2 and Akp6; and both genes are co-expressed in luminal epithelial cells. Studies in cyclic and ovariectomized hamsters established that while progesterone is the major uterine Akp2 inducer, both progesterone and estrogen are strong Akp6 regulators. Studies in preimplantation uteri showed induction of both genes and the activity of their encoded isozymes in luminal epithelial cells during uterine receptivity. However, at the beginning of implantation, Akp2 showed reduced expression in luminal epithelial cells surrounding the implanted embryo. In contrast, expression of Akp6 and its isozyme was maintained in luminal epithelial cells adjacent to, but not away from, the implanted embryo. Following implantation, stromal transformation to decidua was associated with induced expressions of only Akp2 and its isozyme. We next demonstrated that uterine APs dephosphorylate and detoxify endotoxin lipopolysaccharide at their sites of production and activity. Taken together, our findings suggest that uterine APs contribute to uterine receptivity, implantation, and decidualization in addition to their role in protection of the uterus and pregnancy against bacterial infection.

Lei W; Nguyen H; Brown N; Ni H; Kiffer-Moreira T; Reese J; Millan JL; Paria B

2013-08-01

255

Role of lysophosphatidylcholine in brush-border intestinal alkaline phosphatase release and restoration.  

Science.gov (United States)

Intestinal alkaline phosphatase (IAP) is a brush-border membrane ectoenzyme (BBM-IAP) that is released into the lumen (L-IAP) after a high-fat diet. We examined the effects of oil feeding and the addition of mixed-lipid micelles on the formation of L-IAP in oil-fed rat intestine, Caco-2 cell monolayers, and mouse intestinal loops. We localized IAP in the duodenum of rats fed corn oil using fluorescence microscopy with enzyme-labeled fluorescence-97 as substrate. Four hours after oil feeding, L-IAP increased approximately 10-fold accompanied by the loss of BBM-IAP, consistent with BBM-IAP release. Rat IAP isozyme mRNAs progressively increased 4-6 h after oil feeding, followed by the increase of IAP activity in the subapical location at 6 h, consistent with the restoration of IAP protein. Postprandial lipid-micelle components, sodium taurocholate with or without oleic acid, mono-oleylglycerol, cholesterol, or lysophosphatidylcholine (lysoPC) were applied singly or as mixed-lipid micelles to the apical surface of polarized Caco-2 cell monolayers. LysoPC increased L-IAP >10-fold over basal release. LysoPC released IAP into the apical medium more than other intestinal brush-border enzymes, 5'-nucleotidase, sucrase, aminopeptidase N, and lactase, without comparable lactate dehydrogenase release or cell injury. LysoPC increased human IAP mRNA levels by 1.5-fold in Caco-2 cells. Luminally applied lysoPC also increased release of IAP preferentially in mouse intestinal loops. These data show that lysoPC accelerates the formation of L-IAP from BBM-IAP, followed by enhanced IAP synthesis, suggesting the role that lysoPC might play in the turnover of brush-border proteins. PMID:19407215

Nakano, Takanari; Inoue, Ikuo; Alpers, David H; Akiba, Yasutada; Katayama, Shigehiro; Shinozaki, Rina; Kaunitz, Jonathan D; Ohshima, Susumu; Akita, Masumi; Takahashi, Seiichiro; Koyama, Iwao; Matsushita, Makoto; Komoda, Tsugikazu

2009-04-30

256

Characterization of different subpopulations from bone marrow-derived mesenchymal stromal cells by alkaline phosphatase expression.  

UK PubMed Central (United Kingdom)

Multiple surface markers have been utilized for the enrichment of bone marrow mesenchymal stromal cells (MSCs) and to define primitive stem cells. We classified human bone marrow-derived MSC populations according to tissue nonspecific alkaline phosphatase (TNAP) activity. TNAP expression varied among unexpanded primary MSCs, and its level was not related to colony-forming activity or putative surface markers, such as CD105 and CD29, donor age, or gender. TNAP levels were increased in larger cells, and a colony-forming unit-fibroblast assay revealed that the colony size was decreased during in vitro expansion. TNAP-positive (TNAP+) MSCs showed limited multipotential capacity, whereas TNAP-negative (TNAP-) MSCs retained the differentiation potential into 3 lineages (osteogenic-, adipogenic-, and chondrogenic differentiation). High degree of calcium mineralization and high level of osteogenic-related gene expression (osteopontin, dlx5, and cbfa1) were found in TNAP+?cells. In contrast, during chondrogenic differentiation, type II collagen was successfully induced in TNAP- cells, but not in TNAP+?cells. TNAP+?cells showed high levels of the hypertrophic markers, type X collagen and cbfa1. Mesenchymal stem cell antigen-1 (MSCA-1) is identical to TNAP. Therefore, TNAP+?cells were sorted by using antibody targeting MSCA-1. MSCA-1-positive cells sorted for TNAP+?cells exhibited low proliferation rates. Expression of cell cycle-related genes (cyclin A2, CDK2, and CDK4) and pluripotency marker genes (rex1 and nanog) were higher in TNAP- MSC than in TNAP+ MSC. Therefore, TNAP- cells can be described as more primitive bone marrow-derived cells and TNAP levels in MSCs can be used to predict chondrocyte hypertrophy or osteogenic capacity.

Kim YH; Yoon DS; Kim HO; Lee JW

2012-11-01

257

Endotoxin- and ATP-neutralizing activity of alkaline phosphatase as a strategy to limit neuroinflammation.  

UK PubMed Central (United Kingdom)

BACKGROUND: Alkaline phosphatase (AP) is a ubiquitously expressed enzyme which can neutralize endotoxin as well as adenosine triphosphate (ATP), an endogenous danger signal released during brain injury. In this study we assessed a potential therapeutic role for AP in inhibiting neuroinflammation using three complementary approaches. METHODS: Mice were immunized to induce experimental autoimmune encephalomyelitis (EAE) and treated with AP for seven days during different phases of disease. In addition, serological assays to determine AP activity, endotoxin levels and endotoxin-reactive antibodies were performed in a cohort of multiple sclerosis (MS) patients and controls. Finally, the expression of AP and related enzymes CD39 and CD73 was investigated in brain tissue from MS patients and control subjects. RESULTS: AP administration during the priming phase, but not during later stages, of EAE significantly reduced neurological signs. This was accompanied by reduced proliferation of splenocytes to the immunogen, myelin oligodendrocyte glycoprotein peptide. In MS patients, AP activity and isoenzyme distribution were similar to controls. Although endotoxin-reactive IgM was reduced in primary-progressive MS patients, plasma endotoxin levels were not different between groups. Finally, unlike AP and CD73, CD39 was highly upregulated on microglia in white matter lesions of patients with MS. CONCLUSIONS: Our findings demonstrate that: 1) pre-symptomatic AP treatment reduces neurological signs of EAE; 2) MS patients do not have altered circulating levels of AP or endotoxin; and 3) the expression of the AP-like enzyme CD39 is increased on microglia in white matter lesions of MS patients.

Huizinga R; Kreft KL; Onderwater S; Boonstra JG; Brands R; Hintzen RQ; Laman JD

2012-01-01

258

Endotoxin- and ATP-neutralizing activity of alkaline phosphatase as a strategy to limit neuroinflammation  

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Full Text Available Abstract Background Alkaline phosphatase (AP) is a ubiquitously expressed enzyme which can neutralize endotoxin as well as adenosine triphosphate (ATP), an endogenous danger signal released during brain injury. In this study we assessed a potential therapeutic role for AP in inhibiting neuroinflammation using three complementary approaches. Methods Mice were immunized to induce experimental autoimmune encephalomyelitis (EAE) and treated with AP for seven days during different phases of disease. In addition, serological assays to determine AP activity, endotoxin levels and endotoxin-reactive antibodies were performed in a cohort of multiple sclerosis (MS) patients and controls. Finally, the expression of AP and related enzymes CD39 and CD73 was investigated in brain tissue from MS patients and control subjects. Results AP administration during the priming phase, but not during later stages, of EAE significantly reduced neurological signs. This was accompanied by reduced proliferation of splenocytes to the immunogen, myelin oligodendrocyte glycoprotein peptide. In MS patients, AP activity and isoenzyme distribution were similar to controls. Although endotoxin-reactive IgM was reduced in primary-progressive MS patients, plasma endotoxin levels were not different between groups. Finally, unlike AP and CD73, CD39 was highly upregulated on microglia in white matter lesions of patients with MS. Conclusions Our findings demonstrate that: 1) pre-symptomatic AP treatment reduces neurological signs of EAE; 2) MS patients do not have altered circulating levels of AP or endotoxin; and 3) the expression of the AP-like enzyme CD39 is increased on microglia in white matter lesions of MS patients.

Huizinga Ruth; Kreft Karim L; Onderwater Sabina; Boonstra Joke G; Brands Ruud; Hintzen Rogier Q; Laman Jon D

2012-01-01

259

Noninvasive measurement of alkaline phosphatase activity in embryoid bodies and coculture spheroids with scanning electrochemical microscopy.  

Science.gov (United States)

Alkaline phosphatase (ALP) is an enzyme commonly used as an undifferentiated marker of embryonic stem cells (ESCs). Although noninvasive ALP detection has long been desired for stem cell research and in cell transplantation therapy, little progress has been made in developing such techniques. In this study, we propose a noninvasive evaluation method for detecting ALP activity in mouse embryoid bodies (mEBs) using scanning electrochemical microscopy (SECM). SECM has several advantages, including being noninvasive, nonlabeled, quantitative, and highly sensitive. First, we found that SECM-based ALP evaluation permits the comparison of ALP activity among mEBs of different sizes by monitoring the p-aminophenol (PAP) production rate in aqueous solution containing p-aminophenylphosphate (PAPP) normal to the surface area of each sample. Second, coculture spheroids, consisting of mEB and MCF-7 cells for the core and the concentric outer layer, respectively, were prepared as model samples showing heterogeneous ALP activities. The overall PAP production rate dramatically declined in the presence of the MCF-7 cell outer layer, which blocked the mass transfer of PAPP to inner mEB. This result indicated that the SECM response mainly originated from ALP located at the surface of the cellular aggregate, including mEBs and coculture spheroids. Third, taking advantage of the noninvasive nature of SECM, we examined the relevance of ALP activity and cardiomyocyte differentiation. Collectively, these results suggested that noninvasive SECM-based ALP activity normalized by the sample surface enables the selection of EBs with a higher potential to differentiate into cardiomyocytes, which can contribute toward various types of stem cell research. PMID:24053132

Arai, Toshiharu; Nishijo, Taku; Matsumae, Yoshiharu; Zhou, Yuanshu; Ino, Kosuke; Shiku, Hitoshi; Matsue, Tomokazu

2013-10-03

260

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity  

Directory of Open Access Journals (Sweden)

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; 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

2008-01-01

 
 
 
 
261

Cytotoxicity and alkaline phosphatase activity evaluation of endosequence root repair material.  

UK PubMed Central (United Kingdom)

INTRODUCTION: The purpose of this in vitro study was to evaluate the cytotoxicity and alkaline phosphatase (ALP) activity of a new bioceramic root repair material, EndoSequence Root Repair Material (ESRRM; Brasseler USA, Savannah, GA), and to compare these characteristics with those of ProRoot MTA (Dentsply Tulsa Dental, Tulsa, OK) and Geristore (GR; Den-Mat LLC, Santa Maria, CA). METHODS: Human Saos-2 osteoblast-like cells were exposed to 1-, 3-, and 7-day elutes of the materials (100% and 50% strength) for 24 hours after which the bioactivity and ALP activity of the cells were evaluated using a methylthiazol sulfophenyl (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay and para-Nitrophenylphosphate colorimetric assay, respectively. In the positive control group, Triton X-100 (Boehringer Mannheim Corp, Indianapolis, IN) was used to lyse the cells, representing 100% cytotoxicity, and in the negative control group cells received fresh culture medium only. Data were statistically analyzed using the unpaired t test and 1-way analysis of variance. RESULTS: The results revealed that the bioactivity of the cells as well as ALP activity were significantly decreased after exposure to ESRRM elutes in almost all time periods, both in 100% and 50% concentrations, with the exception of ALP activity of day 1 elutes of ESRRM at 50% concentration. MTA did not change the bioactivity or ALP activity of the cells. GR elutes of 100% concentration reduced the bioactivity on days 1 and 3, whereas GR elutes of 50% concentration affected the cells only on day 1. None of the GR elutes had any effect on ALP activity of the cells. CONCLUSIONS: It was concluded that ESRRM elutes of all time periods in general reduced the bioactivity and ALP activity of osteoblast-like cells. GR reduced bioactivity only, whereas MTA had no effect on the cells.

Modareszadeh MR; Di Fiore PM; Tipton DA; Salamat N

2012-08-01

262

Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation  

Scientific Electronic Library Online (English)

Full Text Available 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 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 (more) 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.

2006-05-01

263

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

Scientific Electronic Library Online (English)

Full Text Available Abstract in english 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 f (more) rom 9.5 to 10.5 and Michaelis kinetics for the hydrolysis of p-nitrophenyl phosphate (the Km and Hill coefficient values were 0.35 mM and 1.01, respectively), alpha-naphthyl phosphate (the Km and Hill coefficient values were 0.44 mM and 0.97, respectively), ß-glycerol phosphate (the Km and Hill coefficient values were 2.46 mM and 1.01, respectively) and L-histidinol phosphate (the Km and Hill coefficient values were 0.47 mM and 0.94, respectively) at pH 8.9. The purified enzyme is activated by Mg2+, Zn2+ and Tris-HCl buffer, and is inhibited by Be2+, histidine and EDTA. Also, 0.3 M Tris-HCl buffer protected the purified enzyme against heat inactivation at 70ºC(half-life of 19.0 min, k = 0.036 min-1) as compared to 0.3 M CHES (half-life of 2.3 min, k = 0.392 min-1) in the same experiment.

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

2000-08-01

264

Human Placental Alkaline Phosphatase as a Tracking Marker for Bone Marrow Mesenchymal Stem Cells  

Science.gov (United States)

Abstract Currently, adult mesenchymal stem cells (MSCs) are being evaluated for a wide variety of therapeutic approaches. It has been suggested that MSCs possess regenerative properties when implanted or injected into damaged tissue. However, the efficacy of MSCs in several of the proposed treatments is still controversial. To further explore the therapeutic potential of these cells, it is necessary to trace the fate of individual donor or manipulated cells in the host organism. Recent studies from our lab showed that human placental alkaline phosphatase (hPLAP) is a marker with great potential for cell tracking. However, a potential concern related to this marker is its enzymatic activity, which might alter cell behavior and differentiation by hydrolyzing substrates in the extracellular space and thereby changing the cellular microenvironment. Therefore, the aim of this study was to characterize bone marrow MSCs (BMSCs) derived from hPLAP-transgenic inbred F344 rats (hPLAP-tg) in comparison to wild type (wt) BMSCs. Here, we show that BMSCs from wt and hPLAP-tg donors are indistinguishable in terms of cell morphology, viability, adhesion, immune phenotype, and proliferation as well as in their differentiation capacity over six passages. The expression of the hPLAP marker enzyme was not impaired by extensive in vitro cultivation, osteogenic, adipogenic, or chondrogenic differentiation, or seeding onto two- or three-dimensional biomaterials. Thus, our study underscores the utility of genetically labeled BMSCs isolated from hPLAP-tg donors for long-term tracking of the fate of transplanted MSCs in regenerative therapies.

Balmayor, Elizabeth Rosado; Flicker, Magdalena; Kaser, Tobias; Saalmuller, Armin

2013-01-01

265

Human Placental Alkaline Phosphatase as a Tracking Marker for Bone Marrow Mesenchymal Stem Cells.  

UK PubMed Central (United Kingdom)

Currently, adult mesenchymal stem cells (MSCs) are being evaluated for a wide variety of therapeutic approaches. It has been suggested that MSCs possess regenerative properties when implanted or injected into damaged tissue. However, the efficacy of MSCs in several of the proposed treatments is still controversial. To further explore the therapeutic potential of these cells, it is necessary to trace the fate of individual donor or manipulated cells in the host organism. Recent studies from our lab showed that human placental alkaline phosphatase (hPLAP) is a marker with great potential for cell tracking. However, a potential concern related to this marker is its enzymatic activity, which might alter cell behavior and differentiation by hydrolyzing substrates in the extracellular space and thereby changing the cellular microenvironment. Therefore, the aim of this study was to characterize bone marrow MSCs (BMSCs) derived from hPLAP-transgenic inbred F344 rats (hPLAP-tg) in comparison to wild type (wt) BMSCs. Here, we show that BMSCs from wt and hPLAP-tg donors are indistinguishable in terms of cell morphology, viability, adhesion, immune phenotype, and proliferation as well as in their differentiation capacity over six passages. The expression of the hPLAP marker enzyme was not impaired by extensive in vitro cultivation, osteogenic, adipogenic, or chondrogenic differentiation, or seeding onto two- or three-dimensional biomaterials. Thus, our study underscores the utility of genetically labeled BMSCs isolated from hPLAP-tg donors for long-term tracking of the fate of transplanted MSCs in regenerative therapies.

Balmayor ER; Flicker M; Käser T; Saalmüller A; Erben RG

2013-10-01

266

Effects of Vasectomy on Seminal Plasma Alkaline Phosphatase in Male Alpacas (Vicugna pacos).  

UK PubMed Central (United Kingdom)

Azoospermia is a common finding in male alpacas which present for infertility. The challenge is to differentiate azoospermia of testicular origin from non-testicular origin. In several species, alkaline phosphatase (AP) concentrations in seminal plasma have been used as a diagnostic marker of contributions of the testis and epididymis to the ejaculate. The purpose of this study was to determine whether AP assay could differentiate testicular from non-testicular azoospermia in male alpacas. An experimental model of bilateral outflow obstruction (pre-scrotal vasectomy) was used in 22 male alpacas, aged 2-9 years. No reproductive history was available. Animals were submitted for electroejaculation (EE) under general anaesthesia and vasectomy performed. Five weeks later, animals were submitted for EE. Vasectomy was not successful in one animal, which was removed from analysis. AP levels were compared in seminal plasma in the pre- and post-vasectomy samples. The mean ± SEM concentration of AP in pre-vasectomy seminal plasma was 504.29 ± 166.45 U/l (range 10-2910); the post-vasectomy levels were 252.48 ± 81.77 U/l (range 0-1640; p = 0.06). In 71.4% of animals, AP levels decreased, varying from 18% to 100% reduction. Results of this study suggest that AP is not produced exclusively by the testis and epididymis in alpacas and that AP assay is not a valid diagnostic test for determination of origin of azoospermia; the gold standard for diagnosis of origin of azoospermia remains testicular biopsy.

Pearson L; Campbell A; Sandoval S; Tibary A

2013-06-01

267

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

Directory of Open Access Journals (Sweden)

Full Text Available 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 Michaelis kinetics for the hydrolysis of p-nitrophenyl phosphate (the Km and Hill coefficient values were 0.35 mM and 1.01, respectively), alpha-naphthyl phosphate (the Km and Hill coefficient values were 0.44 mM and 0.97, respectively), ß-glycerol phosphate (the Km and Hill coefficient values were 2.46 mM and 1.01, respectively) and L-histidinol phosphate (the Km and Hill coefficient values were 0.47 mM and 0.94, respectively) at pH 8.9. The purified enzyme is activated by Mg2+, Zn2+ and Tris-HCl buffer, and is inhibited by Be2+, histidine and EDTA. Also, 0.3 M Tris-HCl buffer protected the purified enzyme against heat inactivation at 70ºC(half-life of 19.0 min, k = 0.036 min-1) as compared to 0.3 M CHES (half-life of 2.3 min, k = 0.392 min-1) in the same experiment.

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

2000-01-01

268

Kinetic behaviour of calf intestinal alkaline phosphatase with pNPP.  

UK PubMed Central (United Kingdom)

The hydrolysis of p-nitrophenyl phosphate (pNPP) by calf intestinal alkaline phosphatase (CIAP) was investigated with respect to kinetic parameters such as V(max), K(m) and K(cat) under varying pH, buffers, substrate concentration, temperature and period of incubation. Highest activity was obtained with Tris-HCl at pH 11, while in the case of glycine-NaOH buffer the peak activity was recorded at pH 9.5. The enzyme showed the following kinetic characteristics with pNPP in 50 mM Tris-HCl at pH 11 and 100 mM glycine-NaOH at pH 9.5 at an incubation temperature of 37 degrees C: V(max), 3.12 and 1.6 micromoles min(-1) unit(-1); K(m), 7.6 x 10(-4) M and 4 x 10(-4) M; and K(cat), 82.98 s(-1) and 42.55 s(-1), respectively. CIAP displayed a high temperature optimum of 45 degrees C at pH 11. The kinetic behaviour of the enzyme under different parameters suggested that the enzyme might undergo subtle conformational changes in response to the buffers displaying unique characteristics. Bioprecipitation of Cu2+ from 50 ppm of CuCl2 solution was studied where 64.3% of precipitation was obtained. P(i) generated from CIAP-mediated hydrolysis of pNPP was found to bind with copper and precipitated as copper-phosphate. Thus, CIAP could be used as a test candidate in bioremediation of heavy metals from industrial wastes through generation of metal-phosphate complexes.

Chaudhuri G; Chatterjee S; Venu-Babu P; Ramasamy K; Thilagaraj WR

2013-02-01

269

The toxicity of four native Indian plants: effect on AChE and acid/alkaline phosphatase level in fish Channa marulius.  

UK PubMed Central (United Kingdom)

The latex of four plants viz. Euphorbia royleana, Jatropha gossypifolia (Euphorbiaceae), Nerium indicum and Thevetia peruviana (Apocynaceae) caused significant reduction in acid/alkaline phosphatase activity and anti-acetylcholinesterase activity in nervous tissue of freshwater air breathing fish Channa marulius. The reduction in the activity of both phosphatases and AChE were time as well as dose dependent.

Singh D; Singh A

2005-06-01

270

The toxicity of four native Indian plants: effect on AChE and acid/alkaline phosphatase level in fish Channa marulius.  

Science.gov (United States)

The latex of four plants viz. Euphorbia royleana, Jatropha gossypifolia (Euphorbiaceae), Nerium indicum and Thevetia peruviana (Apocynaceae) caused significant reduction in acid/alkaline phosphatase activity and anti-acetylcholinesterase activity in nervous tissue of freshwater air breathing fish Channa marulius. The reduction in the activity of both phosphatases and AChE were time as well as dose dependent. PMID:15910912

Singh, Digvijay; Singh, Ajay

2005-04-13

271

Membrane-associated alkaline phosphatase from Bacillus licheniformis that requires detergent for solubilization: lactoperoxidase 125I localization and molecular weight determination.  

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When membranes of Bacillus licheniformis MC14 were extracted exhaustively with 1 M magnesium, approximately 80% of the membrane-associated alkaline phosphatase (orthophosphoric-monoester phosphohydrolase [alkaline optimum], E.C. 3.1.3.1) was solubilized. The remaining activity could be extracted wit...

Spencer, D B; Hansa, J G; Stuckmann, K V; Hulett, F M

272

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

UK PubMed Central (United Kingdom)

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.

Zacarias-Soto M; Barón-Sevilla B; Lazo JP

2013-10-01

273

Effect of gingival application of melatonin on alkaline and acid phosphatase, osteopontin and osteocalcin in patients with diabetes and periodontal disease.  

UK PubMed Central (United Kingdom)

Objectives: To assess the effect of topical application of melatonin to the gingiva on salivary fluid concentrations of acid phosphatase, alkaline phosphatase, osteopontin, and osteocalcin. Study Design: Cross-sectional study of 30 patients with diabetes and periodontal disease and 30 healthy subjects. Diabetic patients were treated with topical application of melatonin (1% orabase cream formula) once daily for 20 days and controls with a placebo formulation. Results: Before treatment with melatonin, diabetic patients showed significantly higher mean salivary levels of alkaline and acid phosphatase, osteopontin and osteocalcin than healthy subjects (P < 0.01). After treatment with melatonin, there was a statistically significant decrease of the gingival index (15.84± 10.3 vs 5.6 ± 5.1) and pocket depth (28.3 ± 19.5 vs 11.9 ± 9.0) (P < 0.001). Also, use of melatonin was associated with a significant reduction of the four biomarkers. Changes of salivary acid phosphatase and osteopontin correlated significantly with changes in the gingival index, whereas changes of alkaline phosphatase and osteopontin correlated significantly with changes in the pocket depth. Conclusions: Treatment with topical melatonin was associated with an improvement in the gingival index and pocket depth, a reduction in salivary concentrations of acid phosphatase, alkaline phosphatase, osteopontin and osteocalcin.

Cutando A; López-Valverde A; Gómez-de-Diego R; Arias-Santiago S; de Vicente-Jiménez J

2013-01-01

274

Toxicology of cupric salts on honeybees. V. Gluconate and sulfate action on gut alkaline and acid phosphatases.  

Science.gov (United States)

Some aspects of putative nontarget effects of cupric ions systemically fed to honeybees against their parasite mite Varroa jacobsoni have been investigated on the host phosphatases. The alkaline and acid forms extracted from the guts of worker bees exhibited substrate-inhibition features. Upon detailed kinetic analysis, cupric organic salts indicate activation effects at concentrations of about 1 mM. Concentrations up to 10 mM (alkaline form) and 25 mM (acid form) induced no important changes, except a partial quenching of the substrate-inhibition process, characterized by a wide increase in the constant of apparent inhibitory binding of substrate to the enzyme-substrate complex. Partial purification gave a single alkaline form with quite similar kinetic behavior in the absence of natural ions as in crude extracts. Cupric gluconate and sulfate demonstrated similar patterns, except an increase of the apparent Hill coefficient by sulfate only. The substrate constant of acid phosphatases was decreased at high cupric gluconate doses while its maximum velocity was biphasically increased (with observed maximum at 1 mM), resulting in a sustained activation. Chemiluminescence studies revealed that cupric ion activation is counteracted by oxygen radicals generated by cupric ions and also, in vitro, by the artificial substrate para-nitrophenylphosphate. The para-nitrophenol molecules released from the reaction are therefore responsible for biphasic effects selectively observed with gluconate salts. In apicultural practice, neither blockade of activity nor dramatic changes are to be expected at doses administered to bees against the parasite. PMID:8930506

Bounias, M; Kruk, I; Nectoux, M; Popeskovic, D

1996-10-01

275

Toxicology of cupric salts on honeybees. V. Gluconate and sulfate action on gut alkaline and acid phosphatases.  

UK PubMed Central (United Kingdom)

Some aspects of putative nontarget effects of cupric ions systemically fed to honeybees against their parasite mite Varroa jacobsoni have been investigated on the host phosphatases. The alkaline and acid forms extracted from the guts of worker bees exhibited substrate-inhibition features. Upon detailed kinetic analysis, cupric organic salts indicate activation effects at concentrations of about 1 mM. Concentrations up to 10 mM (alkaline form) and 25 mM (acid form) induced no important changes, except a partial quenching of the substrate-inhibition process, characterized by a wide increase in the constant of apparent inhibitory binding of substrate to the enzyme-substrate complex. Partial purification gave a single alkaline form with quite similar kinetic behavior in the absence of natural ions as in crude extracts. Cupric gluconate and sulfate demonstrated similar patterns, except an increase of the apparent Hill coefficient by sulfate only. The substrate constant of acid phosphatases was decreased at high cupric gluconate doses while its maximum velocity was biphasically increased (with observed maximum at 1 mM), resulting in a sustained activation. Chemiluminescence studies revealed that cupric ion activation is counteracted by oxygen radicals generated by cupric ions and also, in vitro, by the artificial substrate para-nitrophenylphosphate. The para-nitrophenol molecules released from the reaction are therefore responsible for biphasic effects selectively observed with gluconate salts. In apicultural practice, neither blockade of activity nor dramatic changes are to be expected at doses administered to bees against the parasite.

Bounias M; Kruk I; Nectoux M; Popeskovic D

1996-10-01

276

Fluorometric detection of active alkaline phosphatase and gamma-glutamyl transferase in fluid dairy products from multiple species.  

Science.gov (United States)

Over the past 80 years, a variety of methods have been developed to detect underpasteurized or improperly pasteurized milks used in dairy products. Existing methods are hampered by duration of analysis, poor reproducibility, and in some cases the use of hazardous chemicals. To overcome these issues, two new methods have been developed using fluorogenic substrates for two marker enzymes, alkaline phosphatase and gamma-glutamyl transferase. In 30 min, up to 18 samples can be analyzed in triplicate by both methods on two separate 96-well plates. Sample preparation is not necessary for liquid milks when using these methods. The relative standard deviation for each assay is less than 9%, and the correlation coefficient for results of the two methods is greater than 0.98. Using the new methods, milks from four species and nine commercially available liquid milk products were tested. The new methods were also tested directly against an existing phosphatase method (Fluorophos) in spiked whole milk samples. PMID:23643136

Ziobro, George C; McElroy, Kevin M

2013-05-01

277

Fluorometric detection of active alkaline phosphatase and gamma-glutamyl transferase in fluid dairy products from multiple species.  

UK PubMed Central (United Kingdom)

Over the past 80 years, a variety of methods have been developed to detect underpasteurized or improperly pasteurized milks used in dairy products. Existing methods are hampered by duration of analysis, poor reproducibility, and in some cases the use of hazardous chemicals. To overcome these issues, two new methods have been developed using fluorogenic substrates for two marker enzymes, alkaline phosphatase and gamma-glutamyl transferase. In 30 min, up to 18 samples can be analyzed in triplicate by both methods on two separate 96-well plates. Sample preparation is not necessary for liquid milks when using these methods. The relative standard deviation for each assay is less than 9%, and the correlation coefficient for results of the two methods is greater than 0.98. Using the new methods, milks from four species and nine commercially available liquid milk products were tested. The new methods were also tested directly against an existing phosphatase method (Fluorophos) in spiked whole milk samples.

Ziobro GC; McElroy KM

2013-05-01

278

[Effect of cadmium on the activity of alkaline phosphatase (3.1.3.1) of Venus gallina  

UK PubMed Central (United Kingdom)

The marine mollusc Venus gallina was exposed to 32 days sublethal concentrations of Cadmium (0.1 microliter/ml). The activity of alkaline phosphatase was assayed every four days. In the controls the activity was also tested in different tissues and in the soft tissue for the pH dependence. Moreover the influence of direct addition of 10(-4) M Cd on enzyme preparation in vitro was assayed. From the results there are no consistent relationships between the direct in vitro effects of the metal on the enzyme, that is inhibitory, and the effect of exposing the whole animal to the same metal, in this case no inhibition was observed.

Carpenè E; Crisetig G; Cortesi P; Serrazanetti G

1979-07-01

279

Activity of the enzymes of pentose phosphate way and of alkaline phosphatase in rats under incorporation of nitrates and radionuclides  

International Nuclear Information System (INIS)

[en] The pentose-phosphate way major enzymes (transketolase, glucose-6-phosphate dehydrogenase) and the alkaline phosphatase activity in thymus, spleen and mesenteric lymph nodes were studied after introducing of nitrates (5% NaNO3) with drinking water and radionuclides (cesium 137 and strontium 90) with grain during 45-50 days to intact rats and to those suffering B1 hypovitaminosis by injection of oxythiamine. It was revealed changes in the lymphoid tissue cells protein synthesis connected with impairments in the immune mechanisms. The oxythiamine hypovitaminosis caused more marked changes in the enzyme activity in the lymphoid organs

1999-01-01

280

Cell surface-localized alkaline phosphatase of Escherichia coli as visualized by reaction product deposition and ferritin-labeled antibodies.  

UK PubMed Central (United Kingdom)

When cells of a wild-type Eschericia coli O8 strain bearing a complete lipopolysaccharide were incubated for alkaline phosphatase reaction product and examined by electron microscopy, the depostion of lead salts was to be observed primarily within the periplasmic space. A similar treatment of cells derived from this strain, which bears a highly abbreviated lipopolysaccharide, showed a mixed cell surface and periplasmic localization of reaction product, suggesting a surface association of a portion of the enzyme. To further explore this possibility, ferritin-antibody conjugates against the active enzyme and its irreversibly dissociated subunits were prepared and allowed to react with cells of both strains. The results obtained from these experiments revealed the presence of both the active enzyme and inactive subunits of the enzyme at the cell surface of the mutant strain. The evidence obtained offers further proof of the validity of the reaction product deposition technique and indicates that alkaline phosphatase may be associated with some component of the outer membrane in this organism. The observation of enzyme subunits at the cell surface further suggests that an association of these subunits with structural components of the cell envelope may provide a locus at which they may dimerize to form active enzyme.

MacaAlister TJ; Irvin RT; Costerton JW

1977-04-01

 
 
 
 
281

Alkaline phosphatase biosynthesis in the endoplasmic reticulum and its transport through the Golgi apparatus to the plasma membrane: cytochemical evidence.  

UK PubMed Central (United Kingdom)

Enzyme induction of HeLa cell placental alkaline phosphatase with various agents such as prednisolone, sodium butyrate, hyperosmolality (NaCl), or combination of these inducers resulted in the appearance of enzyme activity in the rough endoplasmic reticulum, nuclear envelope, Golgi apparatus, and plasma membrane. In the Golgi apparatus, intense reaction product deposits tended to be concentrated on its trans side, with small vesicles and granules also being positively stained. Inhibition of protein synthesis with cycloheximide was followed by the disappearance of enzyme activity from these cytoplasmic organelles but not from the plasma membrane. Treatment with monensin, a secretory protein transport inhibitor, uniformly increased activity in the rough endoplasmic reticulum while causing marked dilatation of the intensely positive Golgi cisternae. These results suggest that intracellular alkaline phosphatase is newly synthesized in the endoplasmic reticulum and then passes en route through the Golgi apparatus to the plasma membrane. Accordingly, the present system could represent the biosynthesis, transport, and incorporation of the model cell surface enzyme protein to add to the vesicular stomatitus virus glyco-1 (VSV-G) protein and acetylcholine receptor model systems for studying the dynamics of cell surface protein genesis, transport, and membrane integration.

Tokumitsu S; Fishman WH

1983-05-01

282

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

UK PubMed Central (United Kingdom)

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

Jurat-Fuentes JL; Karumbaiah L; Jakka SR; Ning C; Liu C; Wu K; Jackson J; Gould F; Blanco C; Portilla M; Perera O; Adang M

2011-01-01

283

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

Science.gov (United States)

Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP) as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP) were detected by two dimensional differential in-gel electrophoresis (2D-DIGE) analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR) we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests. PMID:21390253

Jurat-Fuentes, Juan Luis; Karumbaiah, Lohitash; Jakka, Siva Rama Krishna; Ning, Changming; Liu, Chenxi; Wu, Kongming; Jackson, Jerreme; Gould, Fred; Blanco, Carlos; Portilla, Maribel; Perera, Omaththage; Adang, Michael

2011-03-01

284

Use of solid phase extraction for the sequential injection determination of alkaline phosphatase activity in dynamic water systems.  

UK PubMed Central (United Kingdom)

In this work, a solid phase extraction sequential injection methodology for the determination of alkaline phosphatase activity in dynamic water systems was developed. The determination of the enzymatic activity was based on the spectrophotometric detection of a coloured product, p-nitrophenol, at 405 nm. The p-nitrophenol is the product of the catalytic decomposition of p-nitrophenyl phosphate, a non-coloured substrate. Considering the low levels expected in natural waters and exploiting the fact of alkaline phosphatase being a metalloprotein, the enzyme was pre-concentrated in-line using a NTA Superflow resin charged with Zn(2+) ions. The developed sequential injection method enabled a quantification range of 0.044-0.441 unit mL(-1) of enzyme activity with a detection limit of 0.0082 unit mL(-1) enzyme activity (1.9 ?mol L(-1) of pNP) and a determination rate of 17 h(-1). Recovery tests confirmed the accuracy of the developed sequential injection method and it was effectively applied to different natural waters and to plant root extracts.

Santos IC; Mesquita RB; Bordalo AA; Rangel AO

2012-08-01

285

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

Energy Technology Data Exchange (ETDEWEB)

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.

Zhang Tingxi [State Key Laboratory of Pollution Control and Resources Reuse, Nanjing University, Nanjing 210093 (China); School of Chemistry and Environmental Science, Nanjing Normal University, Nanjing 210097 (China); Wang Xiaorong [State Key Laboratory of Pollution Control and Resources Reuse, Nanjing University, Nanjing 210093 (China)], E-mail: ekxr@nju.edu.cn; Jin Xiangcan [Research Center of Lake Environment, Chinese Research Academy of Environmental Sciences, Beijing 100012 (China)

2007-11-15

286

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

Energy Technology Data Exchange (ETDEWEB)

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

287

A screen for over-secretion of proteins by yeast based on a dual component cellular phosphatase and immuno-chromogenic stain for exported bacterial alkaline phosphatase reporter.  

UK PubMed Central (United Kingdom)

BACKGROUND: To isolate over-secretors, we subjected to saturation mutagenesis, a strain of P.pastoris exporting E. coli alkaline phosphatase (EAP) fused to the secretory domain of the yeast ? factor pheromone through cellular PHO1/KEX2 secretory processing signals as the ?-sec-EAP reporter protein. Direct chromogenic staining for ?-sec-EAP activity is non-specific as its NBT/BCIP substrate cross-reacts with cellular phosphatases which can be inhibited with Levulinic acid. However, the parental E(P) strain only exports detectable levels of ?-sec-EAP at 69 hours and not within the 36 hour period post-seeding required for effective screening with the consequent absence of a reference for secretion. We substituted the endogenous cellular phosphatase activity as a comparative reference for secretion rate and levels as well as for colony alignment while elevating specificity and sensitivity of detection of the exported protein with other innovative modifications of the immuno-chromogenic staining application for screening protein export mutants. RESULTS: Raising the specificity and utility of staining for ?-sec-EAP activity required 5 modifications including some to published methods. These included, exploitation of endogenous phosphatase activity, reduction of the cell/protein burden, establishment of the direct relation between concentrations of transcriptional inducer and exported membrane immobilized protein and concentrations of protein exported into growth media, amplification of immuno-specificity and sensitivity of detection of ?-sec-EAP reporter enzyme signal and restriction of staining to optimal concentrations of antisera and time periods. The resultant immuno-chromogenic screen allows for the detection of early secretion and as little as 1.3 fold over-secretion of ?-sec-EAP reporter protein by E(M) mutants in the presence of 10 fold -216 fold higher concentrations of HSA. CONCLUSIONS: The modified immuno-chromogenic screen is sensitive, specific and has led to the isolation of mutants E(M) over-secreting the ?-sec-EAP reporter protein by a minimum of 50 fold higher levels than that exported by non-mutagenized E(P) parental strains. Unselected proteins were also over-secreted.

Nallaseth FS; Anderson S

2013-01-01

288

Serum acid phosphatase activities in patients with lung cancer: a biochemical and immunohistochemical analysis of 25 cases.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A series of 25 cases of lung cancer are presented in which total (TAcP) and nonprostatic serum acid phosphatase (NPAcP) activities were measured. Of these cases, 36% had raised TAcP and NPAcP activities in their serum. However, the serum activities of TAcP and NPAcP did not correlate with either the...

Mortimer, G; Casey, M

289

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

290

Production of two extracellular alkaline phosphatases by a psychrophilic Arthrobacter strain  

Energy Technology Data Exchange (ETDEWEB)

We surveyed our collection of psychrophilic bacteria to determine the types of phosphatases they produce and whether any had heat-labile activities with potential applications. Assays at different temperatures showed that the activity from one isolate was optimal at 45{degrees}C and decreased dramatically above 55{degrees}C. This isolate, D10, had the rod-coccus morphological cycle and cell wall amino acids associated with members of the Arthrobacter genus. Interestingly, we found that this strain made two extracellular phosphatases that could be separated by ammonium sulfate fractionation and migration during polyacrylamide gel electrophoresis. One enzyme, designated D10A, hydrolyzed both X-phos (5-bromo-4-chloro-3-indolyl phosphate) and para-nitrophenyl phosphate as substrates and had activity over a broad pH range of 7 to 11. The second enzyme, D10B, lacked activity against X-phos and had a narrow pH range of about 8 to 9. In addition, the D10B enzyme required calcium for activity. The levels of activity of both enzymes decreased for cells grown in media containing more than 100 {mu}M P{sub i}. The results not only demonstrate the existence of different enzymes from one Arthrobacter strain but also suggest ways in which other studies may have missed phosphatases with unknown requirements. 33 refs., 6 figs., 3 tabs.

De Prada, P.; Loveland-Curtze, J.; Brenchley, J.E. [Pennsylvania State Univ., University Park, PA (United States)

1996-10-01

291

[The effects of nicotine on bone calcium and phosphorus content and alkaline phosphatase activity in different part of rat].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To analyze the effects of nicotine on the bone calcium and phosphorus content and alkaline phosphatase (ALP) activity in rat alveolar bone and mandible. METHODS: Twenty health male Wistar rats of five weeks of age were randomly assigned to two groups and received daily intraperitoneal injections for three months as follows: Saline solution for control group, nicotine 0.73 mg.kg-l.d-1 for experimental group. The bone calcium phosphorus content were detected by concentrated acid digestion method and the ALP activity was examined by improved Reddi method. RESULTS: Compared to the control group, bone calcium and phosphorus content was lower in the experimental group (P<0.05), ALP activity had no statistical significance(P>0.05). Bone calcium phosphorus and ALP activity in different parts had no statistical significance (P>0.05). CONCLUSION: The nicotine reduces calcium phosphorus deposition of jaw bone, but has no obvious influence to ALP activity.

Zhao H; Ma S; Chen L; Liu P; Xu J; Hou L; Chen R; Qin C

2013-06-01

292

Changes in bone alkaline phosphatase and procollagen type-1 C-peptide after static and dynamic exercises.  

UK PubMed Central (United Kingdom)

We investigated the effects of two types of nonweight-bearing exercise on changes in bone-specific alkaline phosphatase (BAP) and procollagen type 1 C-peptide (PIP). BAP is a specific marker of bone synthesis, whereas P1P reflects synthesis of type 1 collagen in other organs as well as bone. Eight participants performed static and dynamic unilateral knee extensions. BAP and PIP were measured before, and at 1, 2, 24, 48, and 72 hr after exercise. PIP increased at 24 hr after a static knee extension exercise, whereas BAP did not change during the experimental period. We found no changes in these markers after dynamic exercise. These results imply that type I collagen synthesis in tendons increases after static exercise.

Kubo K; Yuki K; Ikebukuro T

2012-03-01

293

Treatment with bortezomib in multiple myeloma is associated with only a transient and brief increase of bone specific alkaline phosphatase  

DEFF Research Database (Denmark)

There are indications of a bone anabolic effect associated with bortezomib treatment. We present a study with long follow up, measuring bone specific alkaline phosphatase (bALP) for a year during and after treatment in an unselected cohort of myeloma patients treated with bortezomib, and assess factors of potential influence on the increase of bALP. Our main findings are that bALP increase is of short duration and declines significantly even during continued treatment with bortezomib. Only myeloma response was associated with a significant increase of bALP; whereas previous treatment with bortezomib, previous or concomitant treatment with zoledronic acid i.v., dose of bortezomib, line of treatment, or combination with other chemotherapy was not.

Haidl, Felix; Plesner, Torben

2012-01-01

294

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

295

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

Energy Technology Data Exchange (ETDEWEB)

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

296

Cell proliferation and subcellular localization of alkaline phosphatase activity in rat liver parenchyma during azo dye carcinogenesis.  

UK PubMed Central (United Kingdom)

A combined method of phosphatase histochemistry and (3H)thymidine radioautography was devised to study the subcellular localization of alkaline phosphatase (AP) activiity with changing pattern of cell proliferation in precancerous livers of rats fed dimethylaminoazobenzene. After 50 hr of continuous infusion of (3H)thymidine into the rats, labeled liver tissue were fixed in glutaraldehyde. Sections were incubated for AP activity in a lead citrate medium (pH 9.4) with beta-glycerophosphate as substrate. Light and electron microscopic examinations of radioautographs revealed that focal groups of 3H-labeled hepatocytes within hyperplastic nodules were coincident to hyperbasophilic foci and distinguishable from the surrounding parenchyma, which was sparsely labeled. Proliferative hepatocytes in the foci exhibited enzyme reaction product indicative of AP activity along the entire surface membranes. The surface AP tography was in contrast to that of the surrounding hyperplastic parencyma, in which regenerative hepatocytes showed a normal localization of AP activity at the bile canalicular membranes. The L-phenylalanine-sensitive snd heat-resistant activity of hyperbasophilic hepatocytes was different from that of normal hepatocytes. The surface enzyme differentiation was accompanied by a decrease of cytoplasmic AP. Golgi elements apparently function in the mobilization of AP into the surface membranes. The phenomena of AP alterations might be related to the abnormal control of cell proliferation and cytodifferentiation leading to malignant growth.

Karasaki S

1975-03-01

297

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 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 estimada 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 (more) 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 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 (more) 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+.

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

2012-03-01

298

Evidence of Associations Between Feto-Maternal Vitamin D Status, Cord Parathyroid Hormone and Bone-Specific Alkaline Phosphatase, and Newborn Whole Body Bone Mineral Content  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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

Daphna K. Dror; Janet C. King; Ellen B. Fung; Marta D. Van Loan; Erik R. Gertz; Lindsay H. Allen

299

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 h...

Sullip K. Majhi; Shymal Naskar; Binoy K. Mandal

300

Physiological stress induced by sublethal concentrations of phenol and pentachlorophenol in Notopterus Notopterus: hepatic acid and alkaline phosphatases and succinic dehydrogenase  

Energy Technology Data Exchange (ETDEWEB)

The freshwater fish Notopterus notopterus was exposed to four sublethal concentrations (90, 130, 264 and 662 ..mu..g/litre) of phenol and 13.6, 20.4, 40.8 and 60.2 ..mu..g/litre of pentachlorophenol for 10, 20 and 30 days and the effect of these concentrations on the hepatic acid and alkaline phosphatases and succinic dehydrogenase was observed. A significant enzymic difference was found in the liver of control fishes and those exposed to phenol- and pentachlorophenol. The activity of acid and alkaline phosphatases and succinic dehydrogenase was found to be inhibited in almost all cases. The greatest inhibition was observed in acid phosphatase after an exposure period of 30 days. 21 references, 3 tables.

Dalela, R.C.; Rani, S.; Verma, S.R.

1980-01-01

 
 
 
 
301

Exploitation of phosphorescent labelling reagent of fullerol-fluorescein isothiocyanate and new method for the determination of trace alkaline phosphatase as well as forecast of human diseases  

International Nuclear Information System (INIS)

A new phosphorescent labelling reagent consisting of fullerol, fluorescein isothiocyanate and N,N-dimethylaniline (F-ol-(FITC)n-DMA) was developed. The mode of action is based on the reactivity of the active -OH group in F-ol with the -COOH group of FITC to form an F-ol-(FITC)n-DMA complex containing several FITC molecules. F-ol-(FITC)n-DMA increased the number of luminescent molecules in the biological target of WGA-AP-WGA-F-ol-(FITC)n-DMA (WGA and AP are wheat germ agglutinin and alkaline phosphatase, respectively) which improved the sensitivity using solid substrate room temperature phosphorimetry (SSRTP) detection. The proposed method provided high sensitivity and strong specificity for WGA-AP. The limit of detection (LD) was 0.15 ag AP spot-1 for F-ol and 0.097 ag AP spot-1 for FITC in F-ol-(FITC)n-DMA, which was lower than the method using single luminescent molecules of F-ol-DMA and FITC-DMA to label WGA (0.20 ag AP spot-1 for F-ol-DMA and 0.22 ag AP spot-1 for FITC-DMA). Results for the determination of AP in human serum were in good agreement with those obtained by enzyme-linked immunosorbent assay. The mechanism of F-ol-(FITC)n-DMA labelling of WGA was discussed.

2009-08-26

302

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.

Denier Colette C; Brisson-Lougarre Andrée A; Biasini Ghislaine G; Grozdea Jean J; Fournier Didier D

2002-01-01

303

Aplicación del método inmunocitoquímico de la fosfatasa alcalina anti-fosfatasa alcalina para la clasificación inmunológica de las leucemias mieloides agudas/ Application of the alkaline phosphatase anti-alkaline phosphatase immunocytochemical method for the immunological classification of acute myeloid leukemias  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Se realizó el inmunofenotipaje celular de 30 pacientes con el diagnóstico de leucemia mieloide aguda por el método inmunocitoquímico fosfatasa alcalina anti-fosfatasa alcalina (APAAP) introducido en nuestro laboratorio. Los marcadores estudiados fueron: CD3, CD13, CD15, CD19, CD33 y CD41. Para el estudio se utilizaron extendidos de médula ósea o sangre periférica fijados en acetona pura e incubados con el respectivo anticuerpo monoclonal. Posteriormente se añadió (more) la inmunoglobulina anti ratón obtenida en conejo ( Linking ) y por último, el complejo APAAP. Los períodos de incubación fueron de 30 minutos y se realizaron lavados con solución amortiguadora entre cada uno de los pasos. La lectura de las láminas se realizó en microscopio óptico y se consideró positivo cuando el número de células marcadas era mayor o igual a 20 %. De los pacientes estudiados, el 93,3 % y el 90 %, respectivamente, expresaron antígenos pan mieloides CD13 y CD33; 16 de ellos expresaron el CD15 (53,3 %); 3 el CD19 (10 %) y 2 el CD41 (6,6 %). Se concluyó que el método APAAP es rápido y de bajo costo y puede ser aplicado con confiabilidad en la clasificación inmunológica de las leucemias mieloides agudas Abstract in english The cellular immunophenotyping of 30 patients with the diagnosis of acute myeloid leukemia was conducted by the alkaline phosphatase anti-alkaline phosphatase immunocytochemical method (APAAP) introduced in our laboratory. The markers studied were: CD3, CD13, CD15, CD19, CD33 y CD41. Specimens of bone marrow or peripheral blood fixed in pure acetone and incubated with the respective monoclonal antibody were used for the study. Later on, the anti-mouse immunoglobulin obtai (more) ned in rabbit (Linking) was added and, finally, the APAAP complex. The incubation periods were of 30 minutes and lavages with buffer solution were carried out between one step and the other. The reading of the slides was performed on the optical microscope and it was considered positive when the number of marked cells was higher than or equal to 20 %. Of the studied patients, 93.3 % and 90 %, respectively, expressed panmyeloid antigens CD13 and CD33; 16 of them expressed the CD15 (53.3 %); 3 the CD19 (10 %); and 2 the CD41 (6.6 %). It was concluded that the APAAP method is rapid and inexpensive and that it may be reliably applied in the immunological classification of the acute myeloid leukemias

Socarrás Ferrer, Bertha B; del Valle Pérez, Lázaro O; Súarez, Vianed; Merlín Linares, Julio C; Macías Abraham, Consuelo

2006-04-01

304

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

International Nuclear Information System (INIS)

[en] 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

305

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

International Nuclear Information System (INIS)

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

306

Dissolved organic phosphorus utilization and alkaline phosphatase activity of the dinoflagellate Gymnodinium impudicum isolated from the South Sea of Korea  

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This study investigated alkaline phosphatase (APase) activity and dissolved organic and inorganic phosphorus utilization by the harmful dinoflagellate Gymnodinium impudicum (Fraga et Bravo) Hansen et Moestrup isolated from the South Sea of Korea. Under conditions of limited phosphorus, observation of growth kinetics in batch culture yielded a maximum growth rate (?max) of 0.41 /day and a half saturation constant (Ks) of 0.71 ?M. In time-course experiments, APase was induced as dissolved inorganic phosphorus (DIP) concentrations fell below 0.83 ?M, a threshold near the estimated Ks; APase activity increased with further DIP depletion to a maximum of 0.70 pmol/cell/h in the senescent phase. Thus, Ks may be an important index of the threshold DIP concentration for APase induction. G. impudicum utilizes a wide variety of dissolved organic phosphorus compounds in addition to DIP. These results suggest that DIP limitation in the Southern Sea of Korea may have led to the spread of G. impudicum along with the harmful dinoflagellate Cochlodinium polykrikoides in recent years.

Oh, Seok Jin; Kwon, Hyeong Kyu; Noh, Il Hyeon; Yang, Han-Soeb

2010-09-01

307

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

2012-01-01

308

COMPARATIVE ALKALINE PHOSPHATASE CHARACTERISTICS OF THE ALGAL BLOOM DINOFLAGELLATES PROROCENTRUM DONGHAIENSE AND ALEXANDRIUM CATENELLA, AND THE DIATOM SKELETONEMA COSTATUM  

UK PubMed Central (United Kingdom)

The alkaline phosphatase (AP) characteristics of three algal bloom species in the coastal waters of China [Prorocentrum donghaiense D. Lu, Alexandrium catenella (Whedon et Kof.) Balech, and Skeletonema costatum (Grev.) Cleve] were analyzed in a laboratory batch culture experiment using bulk assay and the single-cell enzyme-labeled fluorescence (ELF) method. Results showed that the AP of these three test species shared some common characteristics: AP was inducible in all three species and was expressed by algae under phosphorus (P)-stress conditions; no constitutive AP enzyme was detected in the three test species. Once AP was produced, all three test species gradually released the enzymes into the water, and the algae would reinduce AP production. There were also different specific AP characteristics among the three test species under severe P-stressed conditions. In P. donghaiense, AP covered most of the cell, and the AP production sites were mainly on the cell surface, although some could be observed inside cells. AP also covered the whole cell of A. catenella, but the AP sites were mainly inside the cell with only some on the cell surface. Only one or two AP sites could be detected in S. costatum, and they were all on the cell surface.

Ou Linjian; Huang Bangqin; Hong Huasheng; Qi Yuzao; Lu Songhui

2010-04-01

309

Activities of Aspartate Aminotransferase, Alanine Aminotransferase, Gamma-Glutamyltransferase, Alkaline Phosphatase in Plasma of Postpartum Holstein Cows  

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Full Text Available Depressed appetite and reduced Dry Matter Intake (DMI) and feeding energy-dense diets for a long time around parturition of cows may lead to excessive lipid mobilization which causes the liver damage. This study was meant to determine the effects of postpartun enzymes metabolic status in Holstein cows. In this study, blood samples were during the whole experimental period, obtained from the jugular venepuncture from each animal on 1 week prepartum (week 1), days delivery (week 0) and 1st 9 weeks postpartum (week 1-9). They were analyzed for examining Aspartate aminotransferase (AST), Alanine Aminotransferase (ALT), Gamma-Glutamyltransferase (GGT), Alkaline Phosphatase (ALP) activity. The resultes showed a higher activity of AST which was determined in the 1-3 weeks than other’s. ALT activity indicated a statistically significant increase from the 5-7 weeks of lactation and activity in the 7th week postpartum periods significally reached to the peak. GGT activity in the antepartum 1 week until delivery day was significally lower in comparison with the first to reach the 9th weeks postpartum. ALP activity in the delivery day and 6-8 weeks significant increased in process. Therefore, the AST, ALT, GGT and ALP of enzyme activity which could be used significantly change in the blood plasma of Holstein.

Ping Liu; BaoXiang He; XianLing Yang; XiaoLu Hou; HaiYang Zhao; YinHua Han; Pei Nie; HuiFang Deng; Long Cheng

2012-01-01

310

In vitro response to alkaline phosphatase coatings immobilized onto titanium implants using electrospray deposition or polydopamine-assisted deposition.  

UK PubMed Central (United Kingdom)

Immobilization of biomolecules onto implant surfaces is one of the most straightforward strategies to control the interaction between an implant and its biological environment. Recently, it was shown that the enzyme alkaline phosphatase (ALP) could be efficiently immobilized onto titanium implants in a single step using polydopamine. We hypothesized that such polydopamine-ALP coatings can enhance the early attachment of cells and increase mineralization. Therefore, the current study aimed at immobilization of ALP onto titanium by means of either one- or two-step polydopamine-assisted immobilization or electrospray deposition, the comparative characterization of these experimental substrates and subsequent cell behavioural analysis using primary osteoblast-like cells. Uncoated titanium and ALP-free polydopamine coatings served as controls. Despite significant ALP surface activity and lower water contact for angles ALP-containing surface modifications, only marginal effects on early cell behaviour (i.e. cell spreading) and osteogenic differentiation (i.e. proliferation, differentiation and mineralization) were observed in comparison to uncoated titanium.

Nijhuis AW; van den Beucken JJ; Jansen JA; Leeuwenburgh SC

2013-05-01

311

Cissus quadrangularis extract enhances biomineralization through up-regulation of MAPK-dependent alkaline phosphatase activity in osteoblasts.  

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Cissus quadrangularis Linn. has been implicated as therapeutic agent for enhancing bone healing. Though its osteogenic activity has been suggested, the underlying mechanism still remains unclear. In the present study, the effects of ethanol extract of C. quadrangularis (CQ-E) on osteoblast differentiation and function were analyzed using murine osteoblastic cells. The results indicated that mRNA expressions of osteoblast-related genes were not affected by the CQ-E treatment. However, alkaline phosphatase (ALP) activity and the extent of mineralized nodules were significantly increased in treated cells compared with controls. The addition of an extracellular regulated kinase 1/2 inhibitor, a Jun N-terminal kinase 1/2/3 inhibitor and a p38 mitogen-activated protein kinase (MAPK) inhibitor resulted in significantly decreased ALP activity, preferentially by p38 MAPK inhibitor. These results suggested that CQ-E may regulate osteoblastic activity by enhancing ALP activity and mineralization process, and the increased ALP activity effect of CQ-E is likely mediated by MAPK-dependent pathway. PMID:19057968

Parisuthiman, Duenpim; Singhatanadgit, Weerachai; Dechatiwongse, Thaweephol; Koontongkaew, Sitthichai

2008-12-05

312

Cissus quadrangularis extract enhances biomineralization through up-regulation of MAPK-dependent alkaline phosphatase activity in osteoblasts.  

UK PubMed Central (United Kingdom)

Cissus quadrangularis Linn. has been implicated as therapeutic agent for enhancing bone healing. Though its osteogenic activity has been suggested, the underlying mechanism still remains unclear. In the present study, the effects of ethanol extract of C. quadrangularis (CQ-E) on osteoblast differentiation and function were analyzed using murine osteoblastic cells. The results indicated that mRNA expressions of osteoblast-related genes were not affected by the CQ-E treatment. However, alkaline phosphatase (ALP) activity and the extent of mineralized nodules were significantly increased in treated cells compared with controls. The addition of an extracellular regulated kinase 1/2 inhibitor, a Jun N-terminal kinase 1/2/3 inhibitor and a p38 mitogen-activated protein kinase (MAPK) inhibitor resulted in significantly decreased ALP activity, preferentially by p38 MAPK inhibitor. These results suggested that CQ-E may regulate osteoblastic activity by enhancing ALP activity and mineralization process, and the increased ALP activity effect of CQ-E is likely mediated by MAPK-dependent pathway.

Parisuthiman D; Singhatanadgit W; Dechatiwongse T; Koontongkaew S

2009-03-01

313

Electron-microscopic cytochemistry of alkaline-phosphatase activity in endothelium, pericytes and oligodendrocytes in the rat brain.  

UK PubMed Central (United Kingdom)

The fine structural localization of alkaline-phosphatase (ALP) activity was investigated in the endothelial cells and pericytes of blood vessels and in the oligodendrocytes of rat cerebral cortex and corpus callosum by means of electron-microscopic (EM) cytochemistry using the lead-citrate method. ALP activity was associated with both the luminal and abluminal plasma membranes of some endothelial cells, but in other endothelial cells, this activity was found inside the cytoplasm. In some pericytes, ALP activity was associated with the plasma membrane but in others, strong activity was exhibited within both the cytoplasm and nucleus. Light, medium and dark oligodendrocytes showed ALP activity on their plasma membranes; on the other hand, immature oligodendrocytes exhibited activity within the cytoplasm and on the part of their plasma membrane. Within the cytoplasm of these reacted immature cells, the rough endoplasmic reticulum, nuclear membrane and outer membrane of the mitochondria were the main sites of ALP reaction. Endothelial cells, pericytes and oligodendrocytes demonstrated ALP activity along their plasma membrane or within their cytoplasm, and pericytes showed it within their nuclei. In particular, oligodendrocytes retained ALP activity throughout their cell life, and the intracellular distribution of this activity altered as they matured.

Mori S; Nagano M

1985-01-01

314

Seasonal dynamics, alkaline phosphatase activity and phosphate uptake of adnate and loosely attached epiphytes in an oligotrophic lake  

Energy Technology Data Exchange (ETDEWEB)

This research characterizes the seasonal dynamics of algal epiphytes throughout an annual cycle and examines potential major sources of algal phosphorus. In a phosphorus-limited hardware lake, epiphytic microflora consisted almost entirely of diatoms and blue-green algae, and community structure fluctuated temporally and spatially (with age of macrophyte tissue). Epiphytic ATP was greatest during initial colonization of new macrophyte ramets, and was not correlated with algal biomass except on oldest leaf tissue. Epiphytic chlorophyll was determined primarily by blue-green algal biomass, except on oldest leaf tissue and during colder periods. Epiphytic alkaline phosphatase activity on new and aging macrophyte leaves was significantly less than that on artificial plants, indicating that algae on natural plants were less phosphorus-limited and had obtained a portion of their phosphorus from the macrophytes. Collaborative research quantified the importance of macrophytes as a phosphorus source for algal epiphytes. The author examined the importance of position in determining nutrient availability within the epiphyte matrix. Following short-term assays, scanning electron microscopy (SEM)- and LM-autoradiography were used to directly compare uptake of /sup 33/P-phosphate from the water by intact adnate vs. loosely attached algae, respectively. Loosely attached algae took up significantly more phosphate from the water than did underlying adnate cells, after long-term community acclimation to either phosphorus-poor or phosphorus-enriched conditions.

Burkholder, J.M.

1986-01-01

315

Crevicular Alkaline Phosphatase Activity and Rate of Tooth Movement of Female Orthodontic Subjects under Different Continuous Force Applications.  

UK PubMed Central (United Kingdom)

Purpose. This study is aimed to compare the effects of two different orthodontic forces on crevicular alkaline phosphatase activity, rate of tooth movement, and root resorption. Materials and Methods. Twelve female subjects of class II division 1 malocclusion participated. Maxillary canines with bonded fixed appliances acted as the tested teeth, while their antagonists with no appliances acted as the controls. Canine retraction was performed using nickel titanium coil spring that delivered forces of 100?gm or 150?gm to either side. Crevicular fluid was analyzed for ALP activity, and study models were casted to measure tooth movements. Root resorption was assessed using periapical radiographs before and after the force application. Results. ALP activity at the mesial sites peaked at week 1 for 150?gm group with significant differences when compared with the 100?gm group. Cumulative canine movements were significantly greater in the 150?gm force (2.10 ± 0.50?mm) than in the 100?gm force (1.57 ± 0.44?mm). No root resorption was in the maxillary canines after retraction. Conclusions. A force of 150?gm produced faster tooth movements and higher ALP activity compared with the 100?gm group and had no detrimental effects such as root resorption.

Megat Abdul Wahab R; Md Dasor M; Senafi S; Abang Abdullah AA; Yamamoto Z; Jemain AA; Zainal Ariffin SH

2013-01-01

316

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

Energy Technology Data Exchange (ETDEWEB)

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

317

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

2004-01-01

318

Changes in Expression of Connexin 32, Bile Canaliculus-Like Structures, and Localization of Alkaline Phosphatase in Primary Cultures of Fetal Rat Hepatocytes  

Science.gov (United States)

We devised an experimental design in primary cultures of fetal rat hepatocytes for studying hepatocyte differentiation over a short period. In the present study, hepatocytes were first cultured for 3 days in dexamethasone-supplemented medium and then for an additional 3 days in dexamethasone- or epidermal growth factor-supplemented medium. In hepatocytes cultured continuously in dexamethasone-supplemented medium, the expression of connexin 32 increased and bile canaliculus-like structures and localization of alkaline phosphatase in the plasma membrane around bile canaliculus-like structures were maintained. Few cells incorporated bromodeoxyuridine. On the other hand, in most of the hepatocytes cultured in epidermal growth factor-supplemented medium, the expression of connexin 32 was minimally recognized, bile canaliculus-like structures were shortened or eliminated, and alkaline phosphatase was localized as numerous fine spots throughout the cytoplasm. More than 20% of all hepatocytes incorporated bromodeoxyuridine. The present study suggests that in hepatocytes, there is a close relationship among connexin 32 expression, the maintenance of bile canaliculus-like structures, and the localization of alkaline phosphatase to the plasma membrane around the bile canaliculus-like structures, and this indicates that the present experimental model is useful for studying hepatocyte differentiation over a short period.

Fukazawa, Shoko; Chida, Kohsuke; Taguchi, Meiko; Takeuchi, Akihiro; Ikeda, Noriaki

2013-01-01

319

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; Vianed Marsán Suárez; Miriam Sánchez Segura; Anissa Gramatges Ortiz; Rinaldo Villaescusa Blanco; Consuelo Macías Abraham

2002-01-01

320

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  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish 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 diferenci (more) as estadísticamente significativas para una p Abstract in english 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(more) hough 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

Socarrás Ferrer, Beatriz; Marsán Suárez, Vianed; Sánchez Segura, Miriam; Gramatges Ortiz, Anissa; Villaescusa Blanco, Rinaldo; Macías Abraham, Consuelo

2002-04-01

 
 
 
 
321

Variaciones de la enzima fosfatasa alcalina en la pulpa dental Variations of alkaline phosphatase enzyme in the dental pulp  

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Full Text Available En las últimas décadas, numerosas investigaciones se han dedicado al estudio de los mecanismos potenciales implicados en el desarrollo de la caries dental y su prevención, sin embargo, a pesar de haber disminuido gradualmente el índice de caries en la población, son muchos los pacientes que necesitan tratarse la caries dental, tal es así que continuamente se están utilizando diferentes materiales en la búsqueda de aquel que ante una agresión a la pulpa, ayude a una respuesta biológica de la misma, conservando de esta forma su integridad. De ahí la importancia de la actividad de la fosfatasa alcalina de la pulpa en el proceso carioso, como una reacción ante el hidróxido de calcio que continuamente se está usando en toda la red docente-asistencial del país. Se seleccionaron 50 dientes monorradiculares, con pulpa viva y con caries de segundo, tercer y cuarto grado y 50 dientes sanos de pacientes de diferentes edades. Se extrajo la pulpa de cada diente y se realizaron improntas (3 por cada muestra), una de las cuales se procesó para obtener orientación morfológica, y las otras 2 para valorar la actividad de la fosfatasa alcalina. Para esto se utilizaron 2 métodos: el de calcio cobalto y el de alpha naftol fosfato de Gomori. Como resultado, se obtuvo que la pulpa tiene más actividad enzimática en caries profunda y que la edad del paciente no determina el aumento o disminución de dicha actividad.In the last decades, numerous investigations have been made on the study of potential mechanisms involved in the development of dental caries and their prevention. However, in spite of the gradual reduction of dental caries in the population, a lot of patients need to have their dental caries treated and different materials are continuously used searching for one that before the aggression to the pulp helps it to give a biological response, conserving this way its integrity. That's why the activity of the alkaline phosphatase of the pulp in the caries process is important as a reaction to the calcium hydroxide that is constantly utilized in the teaching-health service network of the country. 50 monoradicular teeth with living pulp and with caries of second, third and fourth degree, and 50 sound teeth from patients of different ages were selected. The pulp of each tooth was extracted and impressions were made (3 per sample). One of them was processed to obtain morphological guidance and the other two to assess the activity of alkaline phosphatase. The cobalt calcium method and Gomori's alpha naphthol phosphate method were used to this end. As a result, it was proved that the pulp has a higher enzymatic activity in deep caries and that the age of the patient does not determine the increase or decrease of this activity.

Zoraida Pons Pinillos; Nadia Hernández Rodríguez

2005-01-01

322

Screening for salivary levels of deoxypyridinoline and bone-specific alkaline phosphatase during orthodontic tooth movement: a pilot study.  

UK PubMed Central (United Kingdom)

Deoxypyridinoline (DPD) and bone-specific alkaline phosphatase (BAP) have been regarded as systemic determinants of bone remodelling. Owing this fact, this study aimed to determine whether the variations in the salivary concentration of these two biomarkers as detected through a longitudinal follow-up with four consecutive visits may be linked with the different phases of orthodontic tooth movement (OTM). Twenty-two healthy subjects who required fixed appliance therapy not involving tooth extractions/surgical procedures were selected. Unstimulated whole saliva samples were collected from each patient prior to fitting the orthodontic appliances and 24-48 hours, 2 weeks, and 5 weeks after the activation. Salivary DPD and BAP concentrations were determined by enzyme-linked immunosorbent assay. The data were analysed using non-parametric statistics. There were no statistically significant differences in salivary levels of biomarkers regarding demographic and clinical parameters. Overall, although DPD values revealed an increasing nature after force application and BAP values showed a descending trend, only the former showed statistically significant changes over time. Furthermore, p ost hoc comparisons for DPD salivary levels revealed significant differences between every paired sampling times, except for the pair baseline test/24-48 hours test. Synchronously, a moderate positive significant correlation between both salivary biomarkers was observed at 2 weeks test. The findings indicate that although salivary levels of DPD and BAP may act as indicators of increased bone remodelling, it appears that DPD dominates the earlier phases of OTM, whereas BAP might serve as indicator of bone formation as soon as the tooth movement stops.

Flórez-Moreno GA; Marín-Restrepo LM; Isaza-Guzmán DM; Tobón-Arroyave SI

2013-06-01

323

1-step versus 2-step immobilization of alkaline phosphatase and bone morphogenetic protein-2 onto implant surfaces using polydopamine.  

Science.gov (United States)

Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the current study, a 1-step polydopamine-based approach was applied for alkaline phosphatase (ALP) and bone morphogenetic protein-2 (BMP-2) immobilization, and compared to the conventional 2-step polydopamine-based immobilization and plain adsorption. To this end, ALP and BMP-2 were immobilized onto titanium and polytetrafluoroethylene (PTFE) substrates. The absolute quantity and biological activity of immobilized ALP were assessed quantitatively to compare the three types of immobilization. Plain adsorption of both ALP and BMP-2 was inferior to both polydopamine-based immobilization approaches. ALP was successfully immobilized onto titanium and PTFE surfaces via the 1-step approach, and the immobilized ALP retained its enzymatic activity. Using the 1-step approach, the amount of immobilized ALP was increased twofold to threefold compared to the conventional 2-step immobilization process. In contrast, more BMP-2 was immobilized using the conventional 2-step immobilization approach. Retention of ALP and BMP-2 was measured over a period of 4 weeks and was found to be similar for the 1-step and 2-step methods and far superior to the retention of adsorbed biomolecules due to the formation of covalent linkages between catechol moieties and immobilized proteins. The biological behavior of ALP and BMP-2 coatings immobilized using polydopamine (1- and 2-step) as well as adsorption was assessed by culturing rat bone marrow cells, which revealed that the cell responses to the various experimental groups were not statistically different. In conclusion, the 1-step polydopamine-based immobilization method was shown to be more efficient for immobilization of ALP, whereas the conventional 2-step method was shown to be more efficient for attachment of BMP-2 onto implant surfaces. PMID:23231507

Nijhuis, Arnold W G; van den Beucken, Jeroen J J P; Boerman, Otto C; Jansen, John A; Leeuwenburgh, Sander C G

2013-01-16

324

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

325

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.)

1997-01-01

326

Repeatability of gingival crevicular fluid collection and quantification, as determined through its alkaline phosphatase activity: implications for diagnostic use.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: In spite of four decades of studies on gingival crevicular fluid, no data have been reported on the repeatability of gingival crevicular fluid collection and the subsequent quantification procedures. The present study reports, for the first time, on the repeatability and method error of gingival crevicular fluid collection and quantification, as determined through its alkaline phosphatase (ALP) activity. Diagnostic considerations are then explored. MATERIAL AND METHODS: Twenty-seven healthy subjects (17 women and 10 men; mean age ± SD, 21.2 ± 4.8 years) with optimal periodontal status were enrolled according to a blind prospective design. The gingival crevicular fluid was collected at baseline, and after 1 d, 1 wk and 3 mo. At each clinical session, two consecutive rounds of gingival crevicular fluid collection were made from each of the four maxillary incisors, allowing the recovery of resting and flow gingival crevicular fluid. The total ALP activities were determined spectrophotometrically, and repeatability and method errors for the resting, flow and overall (resting + flow) gingival crevicular fluid ALP activities were calculated, relative to the corresponding baseline levels. RESULTS: No significant differences were seen over time, although the flow gingival crevicular fluid ALP activity was generally lower than that for the resting gingival crevicular fluid. The method errors ranged from 40 to 58%, with the flow and overall gingival crevicular fluid activities showing the highest and lowest errors, respectively. CONCLUSION: Reliable use of the gingival crevicular fluid ALP collection and quantification, both in research and diagnosis on an individual basis, should take into account relevant errors, and variations are to be considered as true only above relevant thresholds.

Perinetti G; Di Leonardo B; Di Lenarda R; Contardo L

2013-02-01

327

Dephosphorylation of sodium caseinate, enzymatically hydrolyzed casein and casein phosphopeptides by intestinal alkaline phosphatase: implications for iron availability.  

Science.gov (United States)

Clusters of phosphoserine residues in casein bind iron with high affinity. Casein inhibits iron absorption in humans but partial hydrolysis of casein prior to ingestion diminishes this inhibition. The objective of this study was to test two hypotheses: 1. Partial hydrolysis of the peptide bonds in casein exposes phosphoserine residues to attack by intestinal alkaline phosphatase (IAP). 2. Hydrolysis of the phospho-ester linkage in phosphoserine residues in casein by IAP releases bound iron or inhibits iron chelation, thereby allowing its absorption. Test of hypothesis 1: Suspensions of sodium caseinate (SC), enzymatically hydrolyzed casein (EHC), and casein phosphopeptides (CPP) were subjected to an in vitro pepsin/pancreatin digestion and subsequently incubated in the presence of calf IAP. The rate of release of inorganic phosphate was measured with the following results (expressed as &mgr;mol phosphate released/unit of IAP/min): 0.081, 0.104, 0.139 for SC, EHC, and CPP, respectively. These results are consistent with hypothesis 1. Test of hypothesis 2: (59)Fe-citrate or (59)Fe-citrate + CPP in minimum essential media were spiked with a Na(2)WO(4) solution or water (Na(2)WO(4) is a known inhibitor of IAP) and placed on Caco-2 cell monolayers. Uptake of (59)Fe by the cells was used as an index of iron bioavailability. Na(2)WO(4) did not affect (59)Fe uptake from samples containing only iron but did slightly inhibit (by 10%) uptake from samples containing iron + CPP. These results are consistent with hypothesis 2 and provide a possible explanation for the observation that partial hydrolysis of casein improves iron bioavailability. PMID:11382547

Yeung, A C.; Glahn, R P.; Miller, D D.

2001-05-01

328

Dephosphorylation of sodium caseinate, enzymatically hydrolyzed casein and casein phosphopeptides by intestinal alkaline phosphatase: implications for iron availability.  

UK PubMed Central (United Kingdom)

Clusters of phosphoserine residues in casein bind iron with high affinity. Casein inhibits iron absorption in humans but partial hydrolysis of casein prior to ingestion diminishes this inhibition. The objective of this study was to test two hypotheses: 1. Partial hydrolysis of the peptide bonds in casein exposes phosphoserine residues to attack by intestinal alkaline phosphatase (IAP). 2. Hydrolysis of the phospho-ester linkage in phosphoserine residues in casein by IAP releases bound iron or inhibits iron chelation, thereby allowing its absorption. Test of hypothesis 1: Suspensions of sodium caseinate (SC), enzymatically hydrolyzed casein (EHC), and casein phosphopeptides (CPP) were subjected to an in vitro pepsin/pancreatin digestion and subsequently incubated in the presence of calf IAP. The rate of release of inorganic phosphate was measured with the following results (expressed as &mgr;mol phosphate released/unit of IAP/min): 0.081, 0.104, 0.139 for SC, EHC, and CPP, respectively. These results are consistent with hypothesis 1. Test of hypothesis 2: (59)Fe-citrate or (59)Fe-citrate + CPP in minimum essential media were spiked with a Na(2)WO(4) solution or water (Na(2)WO(4) is a known inhibitor of IAP) and placed on Caco-2 cell monolayers. Uptake of (59)Fe by the cells was used as an index of iron bioavailability. Na(2)WO(4) did not affect (59)Fe uptake from samples containing only iron but did slightly inhibit (by 10%) uptake from samples containing iron + CPP. These results are consistent with hypothesis 2 and provide a possible explanation for the observation that partial hydrolysis of casein improves iron bioavailability.

Yeung AC; Glahn RP; Miller DD

2001-05-01

329

1-step versus 2-step immobilization of alkaline phosphatase and bone morphogenetic protein-2 onto implant surfaces using polydopamine.  

UK PubMed Central (United Kingdom)

Immobilization of biomolecules onto implant surfaces is highly relevant in many areas of biomaterial research. Recently, a 2-step immobilization procedure was developed for the facile conjugation of biomolecules onto various surfaces using self-polymerization of dopamine into polydopamine. In the current study, a 1-step polydopamine-based approach was applied for alkaline phosphatase (ALP) and bone morphogenetic protein-2 (BMP-2) immobilization, and compared to the conventional 2-step polydopamine-based immobilization and plain adsorption. To this end, ALP and BMP-2 were immobilized onto titanium and polytetrafluoroethylene (PTFE) substrates. The absolute quantity and biological activity of immobilized ALP were assessed quantitatively to compare the three types of immobilization. Plain adsorption of both ALP and BMP-2 was inferior to both polydopamine-based immobilization approaches. ALP was successfully immobilized onto titanium and PTFE surfaces via the 1-step approach, and the immobilized ALP retained its enzymatic activity. Using the 1-step approach, the amount of immobilized ALP was increased twofold to threefold compared to the conventional 2-step immobilization process. In contrast, more BMP-2 was immobilized using the conventional 2-step immobilization approach. Retention of ALP and BMP-2 was measured over a period of 4 weeks and was found to be similar for the 1-step and 2-step methods and far superior to the retention of adsorbed biomolecules due to the formation of covalent linkages between catechol moieties and immobilized proteins. The biological behavior of ALP and BMP-2 coatings immobilized using polydopamine (1- and 2-step) as well as adsorption was assessed by culturing rat bone marrow cells, which revealed that the cell responses to the various experimental groups were not statistically different. In conclusion, the 1-step polydopamine-based immobilization method was shown to be more efficient for immobilization of ALP, whereas the conventional 2-step method was shown to be more efficient for attachment of BMP-2 onto implant surfaces.

Nijhuis AW; van den Beucken JJ; Boerman OC; Jansen JA; Leeuwenburgh SC

2013-08-01

330

Association of Cry1Ac toxin resistance in Helicoverpa zea (Boddie) with increased alkaline phosphatase levels in the midgut lumen.  

UK PubMed Central (United Kingdom)

Resistance to Bacillus thuringiensis Cry1Ac toxin was characterized in a population of Helicoverpa zea larvae previously shown not to have an alteration in toxin binding as the primary resistance mechanism to this toxin. Cry1Ac-selected larvae (AR1) were resistant to protoxins and toxins of Cry1Ab, Cry1Ac, and the corresponding modified proteins lacking helix ?-1 (Cry1AbMod and Cry1AcMod). When comparing brush border membrane vesicles (BBMVs) prepared from susceptible (LC) and AR1 larval midguts, there were only negligible differences in overall Cry1Ac toxin binding, though AR1 had 18% reversible binding, in contrast to LC, in which all binding was irreversible. However, no differences were detected in Cry1Ac-induced pore formation activity in BBMVs from both strains. Enzymatic activities of two putative Cry1Ac receptors (aminopeptidase N [APN] and alkaline phosphatase [ALP]) were significantly reduced (2-fold and 3-fold, respectively) in BBMVs from AR1 compared to LC larvae. These reductions corresponded to reduced protein levels in midgut luminal contents only in the case of ALP, with an almost 10-fold increase in specific ALP activity in midgut fluids from AR1 compared to LC larvae. Partially purified H. zea ALP bound Cry1Ac toxin in ligand blots and competed with Cry1Ac toxin for BBMV binding. Based on these results, we suggest the existence of at least one mechanism of resistance to Cry1A toxins in H. zea involving binding of Cry1Ac toxin to an ALP receptor in the larval midgut lumen of resistant larvae.

Caccia S; Moar WJ; Chandrashekhar J; Oppert C; Anilkumar KJ; Jurat-Fuentes JL; Ferré J

2012-08-01

331

bFGF and JAGGED1 regulate alkaline phosphatase expression and mineralization in dental tissue-derived mesenchymal stem cells.  

Science.gov (United States)

Basic fibroblast growth factor (bFGF) and Notch signaling play critical roles in various cell behaviors. Here, we investigated the influence of bFGF and Notch signaling in alkaline phosphatase (ALP) expression and mineralization process in human periodontal ligament-derived mesenchymal stem cells (PDLSCs) and stem cells isolated from human exfoliated deciduous teeth (SHEDs). PDLSCs and SHEDs were cultured in osteogenic medium supplemented with bFGF or on the immobilized Notch ligands, JAGGED1. The ALP mRNA and protein expression were measured by quantitative reverse transcriptase polymerase chain reaction and enzymatic activity assay, respectively. Mineral deposition was determined using alizarin red S staining. The results showed that the addition of bFGF resulted in the decrease of ALP mRNA expression and enzymatic activity. In addition, the attenuation of mineralization was noted. These phenomenons were blocked by the addition of a fibroblast growth factor receptor inhibitor (SU5402) or a MEK inhibitor (PD98059). Interestingly, bFGF supplementation also decreased the Notch signaling component mRNA levels. Thus, to evaluate effect of Notch signaling in mineralization process, PDLSCs and SHEDs were exposed to JAGGED1 modified surface. The ALP mRNA and protein expression were significantly upregulated and the mineral deposition was markedly increased. These results could be reversed by the addition of a ?-secretase inhibitor. In addition, bFGF could attenuate the Notch-signaling-induced mineralization in both PDLSCs and SHEDs. These results suggest that mineralization was enhanced by Notch signaling but attenuated by bFGF signaling. This knowledge can be further utilized to control PDLSCs and SHEDs mineralization for tissue regeneration purpose. J. Cell. Biochem. 114: 2551-2561, 2013. © 2013 Wiley Periodicals, Inc. PMID:23749297

Osathanon, Thanaphum; Nowwarote, Nunthawan; Manokawinchoke, Jeeranan; Pavasant, Prasit

2013-11-01

332

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; R Amorim; I Azevedo; M.J Martins

2008-01-01

333

Enhancement of drug delivery to bone: characterization of human tissue-nonspecific alkaline phosphatase tagged with an acidic oligopeptide.  

Science.gov (United States)

Hypophosphatasia is caused by deficiency of activity of the tissue-nonspecific alkaline phosphatase (TNSALP), resulting in a defect of bone mineralization. Enzyme replacement therapy (ERT) with partially purified plasma enzyme was attempted but with little clinical improvement. Attaining clinical effectiveness with ERT for hypophosphatasia may require delivering functional TNSALP enzyme to bone. We tagged the C-terminal-anchorless TNSALP enzyme with an acidic oligopeptide (a six or eight residue stretch of L-Asp), and compared the biochemical properties of the purified tagged and untagged enzymes derived from Chinese hamster ovary cell lines. The specific activities of the purified enzymes tagged with the acidic oligopeptide were the same as the untagged enzyme. In vitro affinity experiments showed the tagged enzymes had 30-fold higher affinity for hydroxyapatite than the untagged enzyme. Lectin affinity chromatography for carbohydrate structure showed little difference among the three enzymes. Biodistribution pattern from single infusion of the fluorescence-labeled enzymes into mice showed delayed clearance from the plasma up to 18 h post infusion and the amount of tagged enzyme retained in bone was 4-fold greater than that of the untagged enzyme. In vitro mineralization assays with the bone marrow from a hypophosphatasia patient using each of the three enzymes in the presence of high concentrations of pyrophosphate provided evidence of bone mineralization. These results show the anchorless enzymes tagged with an acidic oligopeptide are delivered efficiently to bone and function bioactively in bone mineralization, at least in vitro. They suggest potential advantages for use of these tagged enzymes in ERT for hypophosphatasia, which should be explored. PMID:16616566

Nishioka, Tatsuo; Tomatsu, Shunji; Gutierrez, Monica A; Miyamoto, Ken-ichi; Trandafirescu, Georgeta G; Lopez, Patricia L C; Grubb, Jeffrey H; Kanai, Rie; Kobayashi, Hironori; Yamaguchi, Seiji; Gottesman, Gary S; Cahill, Richard; Noguchi, Akihiko; Sly, William S

2006-04-17

334

Enhancement of drug delivery to bone: characterization of human tissue-nonspecific alkaline phosphatase tagged with an acidic oligopeptide.  

UK PubMed Central (United Kingdom)

Hypophosphatasia is caused by deficiency of activity of the tissue-nonspecific alkaline phosphatase (TNSALP), resulting in a defect of bone mineralization. Enzyme replacement therapy (ERT) with partially purified plasma enzyme was attempted but with little clinical improvement. Attaining clinical effectiveness with ERT for hypophosphatasia may require delivering functional TNSALP enzyme to bone. We tagged the C-terminal-anchorless TNSALP enzyme with an acidic oligopeptide (a six or eight residue stretch of L-Asp), and compared the biochemical properties of the purified tagged and untagged enzymes derived from Chinese hamster ovary cell lines. The specific activities of the purified enzymes tagged with the acidic oligopeptide were the same as the untagged enzyme. In vitro affinity experiments showed the tagged enzymes had 30-fold higher affinity for hydroxyapatite than the untagged enzyme. Lectin affinity chromatography for carbohydrate structure showed little difference among the three enzymes. Biodistribution pattern from single infusion of the fluorescence-labeled enzymes into mice showed delayed clearance from the plasma up to 18 h post infusion and the amount of tagged enzyme retained in bone was 4-fold greater than that of the untagged enzyme. In vitro mineralization assays with the bone marrow from a hypophosphatasia patient using each of the three enzymes in the presence of high concentrations of pyrophosphate provided evidence of bone mineralization. These results show the anchorless enzymes tagged with an acidic oligopeptide are delivered efficiently to bone and function bioactively in bone mineralization, at least in vitro. They suggest potential advantages for use of these tagged enzymes in ERT for hypophosphatasia, which should be explored.

Nishioka T; Tomatsu S; Gutierrez MA; Miyamoto K; Trandafirescu GG; Lopez PL; Grubb JH; Kanai R; Kobayashi H; Yamaguchi S; Gottesman GS; Cahill R; Noguchi A; Sly WS

2006-07-01

335

Characterization of eleven novel mutations (M45L, R119H, 544delG, G145V, H154Y, C184Y, D289V, 862+5A, 1172delC, R411X, E459K) in the tissue-nonspecific alkaline phosphatase (TNSALP) gene in patients with severe hypophosphatasia. Mutations in brief no. 217. Online.  

Science.gov (United States)

Hypophosphatasia is a rare inherited disorder characterized by defective bone mineralization and deficiency of serum and tissue liver/ bone/kidney tissue alkaline phosphatase (L/B/K ALP) activity. We report the characterization of tissue-nonspecific alkaline phosphatase (TNSALP) gene mutations in a series of 9 families affected by severe hypophosphatasia. Fourteen distinct mutations were found, 3 of which were previously reported in the North American or Japanese populations. Seven of the 11 new mutations were missense mutations (M45L, R119H, G145V, C184Y and H154Y, D289V, E459K), the four others were 2 single nucleotide deletions (544delG and 1172delC), a mutation affecting donor splice site (862 + 5A) and a nonsense mutation (R411X). PMID:10094560

Taillandier, A; Zurutuza, L; Muller, F; Simon-Bouy, B; Serre, J L; Bird, L; Brenner, R; Boute, O; Cousin, J; Gaillard, D; Heidemann, P H; Steinmann, B; Wallot, M; Mornet, E

1999-01-01

336

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 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 origen (IRDO). Se realizaron las determinaciones de: creatinina, clearence de creatinina (Jaffé cinético), FAL sérica y urinaria (cinético DGKC), microalbuminuria (inmunot (more) urbidimétrico), 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 origin (IRDO). The creatinine, creatinine clearence (kinetic Jaffé) serum and urinary ALP (kinetic DGKC), microalbuminuria (Immunoturbidimetric), proteiunuria (Turbidimetric), ur (more) oproteinogram, 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

2007-09-01

337

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 expe...

A. Zali; A. Nik-Khah; A. Zare Shahneh; K. Rezayazdi; M. Ganjkhanlou

338

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; Miremadi A. Associate Professor; Tabatabaei-e-Yazdi M. Associate Professor; Khodadad-e- Motarjemi M. Resident

2003-01-01

339

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

Directory of Open Access Journals (Sweden)

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; Sudarsan Sabitha; Arun K; Talwar Avaneendra; James Johnson

2009-01-01

340

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 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) h (more) omogenates 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.

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

2008-01-01

 
 
 
 
341

Targeted nucleotide exchange in the alkaline phosphatase gene of HuH-7 cells mediated by a chimeric RNA/DNA oligonucleotide.  

UK PubMed Central (United Kingdom)

Although a variety of methods has been devised for modification of hepatic genes, none has been effective for long-term correction of genetic disorders. In this study, we employed a recently described novel experimental strategy for site-directed nucleotide exchange in genomic DNA of HuH-7 human hepatoma cells. A chimeric 2'-O-methylated-RNA/DNA oligonucleotide containing sequences complementary to 25 bases of the alkaline phosphatase gene was constructed as a duplex containing a G to A substitution at nucleotide 935. Cells were transfected with oligonucleotides for 48 hours, then harvested for DNA isolation and polymerase chain reaction (PCR) amplification of exon 6 of the alkaline phosphatase gene. Colony lifts were hybridized to 17 mer 32P-labeled oligonucleotide probes specific to the 935-G and 935-A sequences. Hybridizing colonies were grown, plasmid DNA isolated, and sequenced. Transfection efficiency was determined at 24 hours by nuclear uptake of fluorescein-12-dUTP-labeled chimeric oligonucleotides. Colonies hybridizing with the 935-A probe were identified only from cells transfected with the specific chimeric oligonucleotide; and there was no evidence of cross-hybridization. Conversion of G to A at nucleotide 935 occurred at an overall frequency of up to 11.9% and when corrected for transfection efficiency approached 43%. No other alterations were detected in the sequence of exon 6 with the targeted nucleotide exchange. These results show that a single base pair alteration in the alkaline phosphatase gene of HuH-7 cells can be introduced at a relatively high frequency following transfection with chimeric RNA/DNA oligonucleotides. This technique offers a novel and potentially powerful strategy for site-directed hepatic gene alteration without the use of viral-based vectors.

Kren BT; Cole-Strauss A; Kmiec EB; Steer CJ

1997-06-01

342

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

343

CdS/TiO2-fluorescein isothiocyanate nanoparticles as fluorescence resonance energy transfer probe for the determination of trace alkaline phosphatase based on affinity adsorption assay.  

Science.gov (United States)

The CdS/TiO(2)-fluorescein isothiocyanate (FITC) luminescent nanoparticles (CdS/TiO(2)-FITC) with the particle size of 20 nm have been synthesized by sol-gel method. CdS/TiO(2)-FITC could emit the fluorescence of both FITC and CdS/TiO(2). The fluorescence resonance energy transfer (FRET) occurred between the donor CdS/TiO(2) and the acceptor FITC in the CdS/TiO(2)-FITC. Taking advantages of the excellent characteristics of FRET, a new CdS/TiO(2)-FITC FRET labeling reagent and a CdS/TiO(2)-FITC-wheat germ agglutinin (CdS/TiO(2)-FITC-WGA) fluorescent probe have been developed. The FRET occurring between the donor CdS/TiO(2) and the acceptor FITC in the labelled product CdS/TiO(2)-FITC-WGA-AP, formed in the affinity adsorption reaction between the WGA in this CdS/TiO(2)-FITC-WGA fluorescent probe and alkaline phosphatase (AP), sharply enhanced the fluorescence signal of FITC and quench the fluorescence signal of CdS/TiO(2). Moreover, the ?F (the change of the fluorescence signal) of FITC and CdS/TiO(2) were proportional to the content of AP, respectively. Thus, a new method that CdS/TiO(2)-fluorescein isothiocyanate nanoparticles for the determination of trace AP based on FRET-affinity adsorption assay has been established. The limit of quantification (LOQ) of the method was 1.3×10(-17) g AP mL(-1) for CdS/TiO(2) and 1.1×10(-17) g AP mL(-1) for FITC, respectively. This sensitive, rapid, high selective and precise method has been applied to the determination of AP in human serum and the prediction of human disease with the results agreed well with enzyme-linked immunosorbent assay (ELISA) in Zhangzhou Municipal Hospital of Fujian Province. Simultaneously, the reaction mechanism for the determination of AP was also discussed. PMID:22939139

Liu, Jia-Ming; Lin, Li-ping; Jiao, Li; Cui, Ma-Lin; Wang, Xin-Xing; Zhang, Li-Hong; Zheng, Zhi-Yong

2012-06-29

344

Relationship between bone formation markers bone alkaline phosphatase, osteocalcin and amino-terminal propeptide of type I collagen and bone mineral density in elderly men. Preliminary results.  

Science.gov (United States)

Bone remodeling is altered in all metabolic bone diseases, especially in post-menopausal women and in the elderly. Predicting changes in bone mineral density (BMD) is useful to manage the progression of such diseases and to potentially provide interventions in reducing fracture risk. Continuous bone formation and resorption processes can be monitored by measuring biochemical markers of bone turnover (BTMs) and a relationship between BMD and BTMs has been known for long. The aim of this study was to evaluate the relationship between BMD and serum BTMs bone alkaline phosphatase (BAP), osteocalcin and amino-terminal propeptide of type I collegen (PINP) in elderly (>65 years) men. We prospectively studied 18 elderly men (median age=69, range=65-77 years) with no history of fractures, angina, stroke, myocardial infarction or diabetes mellitus. Patients who had undergone corticosteroid, calcitonin, androgen or bisphosphonate therapy were excluded from the study, as well as those who were vitamin D and calcium supplementation users. All the patients underwent lumbar-spine (L2-L4) dual-energy x-ray absorbtiometry and BMD, BAP, osteocalcin and PINP measurements. The mean BMD and body mass index (BMI) were 0.963±0.04 g/cm(2) and 24.4±1.2 kg/m(2), respectively. BAP, osteocalcin and PINP were 27.8±11.3 U/l, 25.6±7.1 ng/ml and 36.0±7.5 ng/ml, respectively. No correlation was found between BMD and BAP (R=-0.28, p=0.25), osteocalcin (R=-0.18, p=0.48) and PINP (R=-0.21, p=0.39), nor between BMI and both age (R=0.05, p=0.83) and BMD (R=0.10, p=0.67). In conclusion, we did not find any relationship between bone formation markers BAP, osteocalcin and PINP and bone density. Thus, our preliminary data suggest that BTMs are not useful in monitoring the bone mineral status of elderly men. PMID:23160690

Lumachi, Franco; Orlando, Rocco; Fallo, Francesco; Basso, Stefano M M

345

Relationship between bone formation markers bone alkaline phosphatase, osteocalcin and amino-terminal propeptide of type I collagen and bone mineral density in elderly men. Preliminary results.  

UK PubMed Central (United Kingdom)

Bone remodeling is altered in all metabolic bone diseases, especially in post-menopausal women and in the elderly. Predicting changes in bone mineral density (BMD) is useful to manage the progression of such diseases and to potentially provide interventions in reducing fracture risk. Continuous bone formation and resorption processes can be monitored by measuring biochemical markers of bone turnover (BTMs) and a relationship between BMD and BTMs has been known for long. The aim of this study was to evaluate the relationship between BMD and serum BTMs bone alkaline phosphatase (BAP), osteocalcin and amino-terminal propeptide of type I collegen (PINP) in elderly (>65 years) men. We prospectively studied 18 elderly men (median age=69, range=65-77 years) with no history of fractures, angina, stroke, myocardial infarction or diabetes mellitus. Patients who had undergone corticosteroid, calcitonin, androgen or bisphosphonate therapy were excluded from the study, as well as those who were vitamin D and calcium supplementation users. All the patients underwent lumbar-spine (L2-L4) dual-energy x-ray absorbtiometry and BMD, BAP, osteocalcin and PINP measurements. The mean BMD and body mass index (BMI) were 0.963±0.04 g/cm(2) and 24.4±1.2 kg/m(2), respectively. BAP, osteocalcin and PINP were 27.8±11.3 U/l, 25.6±7.1 ng/ml and 36.0±7.5 ng/ml, respectively. No correlation was found between BMD and BAP (R=-0.28, p=0.25), osteocalcin (R=-0.18, p=0.48) and PINP (R=-0.21, p=0.39), nor between BMI and both age (R=0.05, p=0.83) and BMD (R=0.10, p=0.67). In conclusion, we did not find any relationship between bone formation markers BAP, osteocalcin and PINP and bone density. Thus, our preliminary data suggest that BTMs are not useful in monitoring the bone mineral status of elderly men.

Lumachi F; Orlando R; Fallo F; Basso SM

2012-11-01

346

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.

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

2010-01-01

347

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

UK PubMed Central (United Kingdom)

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.

Stavri S; Zarnescu O

2013-02-01

348

Intestinal alkaline phosphatase activity as a molecular marker of enterotoxicity induced by single dose of 5-fluorouracil and protective role of orally administered glutamine  

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Full Text Available Background. One of the critical limitations for the administration of the chemotherapy is the toxicity affecting normal tissue. The main target organs for 5-fluorouracil (5-FU) toxicity in humans and experimental animals are the gastrointestinal tract, bone marrow, and skin. The cytotoxic effects of antimetabolite chemotherapy are based on their role as substrates for the same transport processes and enzymes involved in anabolism and catabolism as the natural substrates. The main goal of our study was to analyze the dose-dependent antiproliferative effects of 5-FU on intestinal mucosa, enterotoxic potential of 5-FU in experimental animals and to test possible protective role of glutamine. Methods. In our study, we used Sprague Dawley rats. The control group of rats included 50 animals, while the groups where either 5-fluorouracil (5-FU) alone or 5-FU and glutamine were administered included 200 animals. All experimental animals were further stratified according to the experimental model (25 animals in each of 8 experimental subgroups of animals). The 5-FU was administered by intraperitoneal application in single dose of 0, 100, 200, 300, and 400 mg of 5-FU per kg of body weight. Water solution of 1% glutamine was prepared daily and administered orally, in volume of 200 ml, for 7 days continuously, after the 7th day of 5-FU administration. Experimental animals were sacrificed 7 days after the administration of 5-FU. The isolation of enterocytes was performed according to the method of Kralovansky et al. In cell homogenate obtained by described method, we determined the protein content using the Biuret method and the DNA content using the Burton reagent. The activities of enzymes alkaline phosphatase (ALP), glutathione S-transferase (GST), glutathione reductase (GR), and glutathione peroxidase (GPX) were determined by kinetic method. All paraffin samples of the small intestine were stained by haematoxiline and eosine(HE method). All the experiments were done in duplicate and analyzed by standard statistical methods. All the experiments were done in duplicate and analyzed by standard statistical methods. Results: Our results of enterotoxicity induced by intraperitonealy administered 5-FU showed statistically significant decrease of DNA content in small intestine samples of experimental animals, decrease in activity of intestinal alkaline phosphatase enzyme and the increase in glutathione-dependent enzymes. The glutamine supplementation reduced 5-FU intestinal toxicity. Conclusion: Intestinal alkaline phosphatase is a good marker of the dose-dependent enterotoxicity induced by 5-fluorouracil.

Bajin-Kati? Katica; Stankov Karmen; ?olai Matilda; Kova?evi? Zoran

2006-01-01

349

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

UK PubMed Central (United Kingdom)

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.

Urquiza NM; Manca SG; Moyano MA; Dellmans RA; Lezama L; Rojo T; Naso LG; Williams PA; Ferrer EG

2010-04-01

350

Ferrocenylethyl phosphate:? an improved substrate for the detection of alkaline phosphatase by cathodic stripping ion-exchange voltammetry. Application to the electrochemical enzyme affinity assay of avidin.  

UK PubMed Central (United Kingdom)

The ferrocenylethyl phosphate disodium salt was synthesized and used as a new substrate for alkaline phosphatase (AP). The enzyme-generated ferroceneethanol was selectively and sensitively detected at a Nafion film-coated electrode by anodic preconcentration of the ferricinium salt, followed by cyclic voltammetry. The accumulated ferricinium units could be expelled from the polymer film in their neutral form by cathodic stripping, and so the Nafion-modified electrode could be reused for more than 10 measurements with a standard deviation less than 3%. Values of 0.75 mM for the Michaelis constant and 1.42 ?mol s(-)(1) (mg of protein)(-)(1) for the maximal velocity were found. The regenerable Nafion-coated electrode was employed for the indirect detection of AP down to 2 × 10(-)(12) M and for the noncompetitive heterogeneous enzyme assay of avidin, whose detection limit was 5 × 10(-)(12) M.

Limoges B; Degrand C

1996-12-01

351

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

UK PubMed Central (United Kingdom)

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.

Zúñiga-Navarrete F; Gómez I; Peña G; Bravo A; Soberón M

2013-03-01

352

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

2012-06-26

353

Quantitation of non-amplified genomic DNA by bead-based hybridization and template mediated extension coupled to alkaline phosphatase signal amplification.  

Science.gov (United States)

Klenow I polymerase activity was combined with solid phase DNA hybridization to detect non-amplified genomic DNA (gDNA) sequences from Escherichia coli. Aminopropyl-controlled pore glass surface-bound oligonucleotides were hybridized to fragmented gDNA. The template-mediated extension at the 3'-terminus of the immobilized probe was then promoted in the presence of Klenow I polymerase and digoxigenin-labeled nucleotides. Detection of the extended probes was accomplished with an anti-digoxigenin alkaline phosphatase conjugate protocol coupled to colorimetric or fluorescent detection. Using the colorimetric protocol, the proof-of-concept was established. The fluorescence-based methodology, on the other hand, provided the basis for a quantitative interpretation of the data, affording a detection limit of 5 pM gDNA. PMID:19838631

Martins, S A M; Prazeres, D M F; Fonseca, L P; Monteiro, G A

2009-10-17

354

Quantitation of non-amplified genomic DNA by bead-based hybridization and template mediated extension coupled to alkaline phosphatase signal amplification.  

UK PubMed Central (United Kingdom)

Klenow I polymerase activity was combined with solid phase DNA hybridization to detect non-amplified genomic DNA (gDNA) sequences from Escherichia coli. Aminopropyl-controlled pore glass surface-bound oligonucleotides were hybridized to fragmented gDNA. The template-mediated extension at the 3'-terminus of the immobilized probe was then promoted in the presence of Klenow I polymerase and digoxigenin-labeled nucleotides. Detection of the extended probes was accomplished with an anti-digoxigenin alkaline phosphatase conjugate protocol coupled to colorimetric or fluorescent detection. Using the colorimetric protocol, the proof-of-concept was established. The fluorescence-based methodology, on the other hand, provided the basis for a quantitative interpretation of the data, affording a detection limit of 5 pM gDNA.

Martins SA; Prazeres DM; Fonseca LP; Monteiro GA

2010-02-01

355

Comparison of Salivary Calcium, Phosphate and Alkaline Phosphatase in Children with Severe, Moderate caries and Caries Free in Tehran’s Kindergarthens  

Directory of Open Access Journals (Sweden)

Full Text Available Background: The most common dental disease in childhood is dental caries. This study was carried out to recognize the components of saliva which are protective factors in children in order to evaluate and predict caries susceptible and caries resistant individuals. Methods: A total of 75 subjects of either sex aged 3-5 years old from kindergartens in Tehran were selected and divided into 3 groups (case group: dmft>6, control group 1: 10.05). However, the results showed that salivary phosphate and alkaline phosphatase in caries free group and calcium in the group with severe caries was somewhat more than those in other groups. Conclusions: Despite the results of the present study, the relationship between salivary components and caries rate in children remainslcontroversial. So, more and wide studies are necessary to achieve some practical criteria for predicting dental caries, recognition of susceptible persons and finally prevention of caries in children.

M Shahrabi; B Seraj; MT HaghiAshtiani; N Akhundi; A Alikhani

2007-01-01

356

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

DEFF Research Database (Denmark)

Immuno-alkaline phosphatase (AP) staining for T cell subsets (CD4 and CD8) of smears from fingerprick blood functioned well under tropical climatic conditions when smears were stored frozen with silica gel before being labelled. Unlabelled smears were stored for up to 12 months and could be transferred abroad without antigenic damage. Identical total CD4 and CD8 counts were obtained on venous and capillary blood, when compared using a FACS analyser. Although the AP method gave somewhat higher total CD4 and CD8 counts, the ratio remained the same. The major advantages of the method are: (i) no expensive equipment is required, (ii) only minute amounts of blood are needed, and (iii) slides can be stored for long periods before labelling and can be preserved for later reading. The method is suitable for community studies where there is a need for assessing the immune status of the population.

Lisse, I M; Whittle, H

1990-01-01

357

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

Directory of Open Access Journals (Sweden)

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; Cañizares M E; Curbelo A; González B; Rivero Y; Thomas A

2008-01-01

358

Correlation between serum enzymes and serum unsaturated vitamin B12 binding proteins in primary liver carcinoma.  

UK PubMed Central (United Kingdom)

The serum unsaturated vitamin B12-binding capacity (UBBC), unsaturated transcobalamin (UTC) I, UTC II, UTC III levels, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase activities and bilirubin concentration were estimated in 61 patients with liver diseases (31 with hepatoma, 30 with viral hepatitis). The levels of serum cobalamin, UTC I, UTC III, UBBC, alanine and aspartate aminotransferases, and bilirubin were raised in both hepatoma and viral hepatitis patients. Serum UTC II was reduced in both conditions. Alkaline phosphatase activity was significantly increased in hepatoma. Four significant correlations were observed among these parameters in the hepatoma patients while only one significant correlation was observed in viral hepatitis.

Osifo BO; Ayoola A; Parmentier Y; Gerard P; Nicolas JP

1988-01-01

359

[Transient alkaline hyperphosphatemia of childhood. Description of a clinical case].  

Science.gov (United States)

Transient idiopathic alkaline hyperphosphatasemia is a syndrome of unknown etiology unrelated to any specific disease, characterized by a marked transient increase of the serum level of alkaline phosphatase. About fifty-two cases of them included from thirty-seven to two months have been reported in Literature till now. A new case of a little girl four years and three months old is reported here. PMID:3328161

Dello Iacono, I; Tomaselli, D E; Quarantiello, F; Vicario, V; Sellitto, F

360

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

Energy Technology Data Exchange (ETDEWEB)

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

 
 
 
 
361

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

DEFF Research Database (Denmark)

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. Udgivelsesdato: 1986-Jun-1

Honoré, B; Frandsen, P C

1986-01-01

362

Antibody guided diagnosis and therapy of brain gliomas using radiolabeled monoclonal antibodies against epidermal growth factor receptor and placental alkaline phosphatase  

Energy Technology Data Exchange (ETDEWEB)

Twenty-seven patients with brain glioma were scanned using {sup 123}I-labeled monoclonal antibodies against epidermal growth factor receptor (EGFR1) or placental alkaline phosphatase (H17E2). Successful localization was achieved in 18 out of 27 patients. Eleven out of 27 patients were also studied using a nonspecific control antibody (11.4.1) of the same immunoglobulin subclass and observable tumor localization was also achieved in five patients. The specificity of targeting was assessed by comparing images obtained with specific and nonspecific antibodies and by examining tumor and normal tissue biopsies after dual antibody administration. Ten patients with recurrent grade III or IV glioma who showed good localization of radiolabeled antibody were treated with 40-140 mCi of {sup 131}I-labeled antibody delivered to the tumor area intravenously (n = 5) or by infusion into the internal carotid artery (n = 5). Six patients showed clinical improvement lasting from 6 mo to 3 yr. One patient continues in remission (3 yr after therapy), but the other five who responded initially relapsed 6-9 mo after therapy and died. No major toxicity was attributable to antibody-guided irradiation. Targeted irradiation by monoclonal antibody may be clinically useful and should be explored further in the treatment of brain gliomas resistant to conventional forms of treatment.

Kalofonos, H.P.; Pawlikowska, T.R.; Hemingway, A.; Courtenay-Luck, N.; Dhokia, B.; Snook, D.; Sivolapenko, G.B.; Hooker, G.R.; McKenzie, C.G.; Lavender, P.J. (Royal Postgraduate Medical School, London (England))

1989-10-01

363

Antibody guided diagnosis and therapy of brain gliomas using radiolabeled monoclonal antibodies against epidermal growth factor receptor and placental alkaline phosphatase  

International Nuclear Information System (INIS)

Twenty-seven patients with brain glioma were scanned using 123I-labeled monoclonal antibodies against epidermal growth factor receptor (EGFR1) or placental alkaline phosphatase (H17E2). Successful localization was achieved in 18 out of 27 patients. Eleven out of 27 patients were also studied using a nonspecific control antibody (11.4.1) of the same immunoglobulin subclass and observable tumor localization was also achieved in five patients. The specificity of targeting was assessed by comparing images obtained with specific and nonspecific antibodies and by examining tumor and normal tissue biopsies after dual antibody administration. Ten patients with recurrent grade III or IV glioma who showed good localization of radiolabeled antibody were treated with 40-140 mCi of 131I-labeled antibody delivered to the tumor area intravenously (n = 5) or by infusion into the internal carotid artery (n = 5). Six patients showed clinical improvement lasting from 6 mo to 3 yr. One patient continues in remission (3 yr after therapy), but the other five who responded initially relapsed 6-9 mo after therapy and died. No major toxicity was attributable to antibody-guided irradiation. Targeted irradiation by monoclonal antibody may be clinically useful and should be explored further in the treatment of brain gliomas resistant to conventional forms of treatment.

1989-01-01

364

Single domain antibody-alkaline phosphatase fusion proteins for antigen detection--analysis of affinity and thermal stability of single domain antibody.  

Science.gov (United States)

Single domain antibody (sdAb)-alkaline phosphatase (AP) fusion proteins have been demonstrated to be useful immunodiagnostic reagents for bio-threat agent detection. The bivalent nature of sdAb-AP fusion proteins significantly increases effective affinity and thus the sensitivity of detection, but the thermal stability of the fusion protein had not been explored. This property is critical for the development of immunoassays for use in austere environments. In this study four sdAbs with specificity for MS2 phage coat protein (CP) were expressed as fusions with AP in order to evaluate the thermal stability and affinity of the resulting constructs. The melting temperature (Tm) of the sdAb and sdAb-AP fusion proteins was measured by a combination of Circular Dichroism (CD), differential scanning calorimetry (DSC) and Fluorescence-based Thermal Shift assay. Binding kinetics were assessed using surface plasmon resonance (SPR). Our results indicated that the AP fusion protein did not increase the Tm or enhance thermal stability of the sdAb, but did provide the expected increase in binding affinity as compared to the original sdAb. PMID:23570946

Liu, Jinny L; Zabetakis, Dan; Lee, Audrey Brozozog; Goldman, Ellen R; Anderson, George P

2013-04-06

365

The zonal distributions of alkaline phosphatase, adenosine triphosphatase, laminin, fibronectin and chondroitin 4-sulphate in growing rat humerus proximal epiphyseal cartilage: a histochemical and an immunohistochemical study.  

Science.gov (United States)

Although there are many studies about epiphyseal cartilage extracellular matrix (ECM) macromolecules in bone formation, studies of their distribution and role in the mineralization of these components in growing rat humerus proximal epiphyseal cartilage have not been sufficiently detailed. The aim of this study was to determine the distributions of alkaline phosphatase (ALP), adenosine triphosphatase (ATPase), laminin, fibronectin and chondroitin 4-sulphate in growing rat humerus proximal epiphyseal cartilage. The rats were killed by cervical dislocation, and the humeri were removed, sectioned (6 and 10 microm) on a cryotome or paraffin microtome, and stained using histochemical and immunohistochemical methods. ALP and ATPase were markedly observed in the hypertrophy and calcifying cartilage. In addition, ATPase was found to be very strongly positive in the tangential zone of articular cartilage. Results of immunohistochemical staining for laminin, fibronectin and chondroitin 4-sulphate showed that the immunostaining was the heaviest in the tangential zone of articular cartilage. In growing epiphyseal plates, there were differences in the density of these macromolecules of chondrocytes as a function of the maturation process. In conclusion, these ECM macromolecules of epiphyseal cartilage may regulate the cell-cell and cell-matrix interactions as well as the matrix calcification during the ossification of epiphyseal cartilage. PMID:14651483

Ustünel, I; Sahin, Z; Akkoyunlu, G; Demir, R

2003-12-01

366

Single domain antibody-alkaline phosphatase fusion proteins for antigen detection--analysis of affinity and thermal stability of single domain antibody.  

UK PubMed Central (United Kingdom)

Single domain antibody (sdAb)-alkaline phosphatase (AP) fusion proteins have been demonstrated to be useful immunodiagnostic reagents for bio-threat agent detection. The bivalent nature of sdAb-AP fusion proteins significantly increases effective affinity and thus the sensitivity of detection, but the thermal stability of the fusion protein had not been explored. This property is critical for the development of immunoassays for use in austere environments. In this study four sdAbs with specificity for MS2 phage coat protein (CP) were expressed as fusions with AP in order to evaluate the thermal stability and affinity of the resulting constructs. The melting temperature (Tm) of the sdAb and sdAb-AP fusion proteins was measured by a combination of Circular Dichroism (CD), differential scanning calorimetry (DSC) and Fluorescence-based Thermal Shift assay. Binding kinetics were assessed using surface plasmon resonance (SPR). Our results indicated that the AP fusion protein did not increase the Tm or enhance thermal stability of the sdAb, but did provide the expected increase in binding affinity as compared to the original sdAb.

Liu JL; Zabetakis D; Lee AB; Goldman ER; Anderson GP

2013-07-01

367

Trichostatin A, an HDAC class I/II inhibitor, promotes pi-induced vascular calcification via up-regulation of the expression of alkaline phosphatase.  

UK PubMed Central (United Kingdom)

AIM: Vascular calcification, a major complication of chronic kidney disease (CKD), refers to the mineralization of vascular smooth muscle cells (VSMCs), resulting from a phenotypic change towards osteoblast-like cells. Histone deacetylase inhibitors (HDIs), potential therapeutic agents for CKD, are known to promote the differentiation and mineralization of osteoblasts. In this study, we aimed to determine the effects of an HDI on the phenotypic change of VSMCs and the development of vascular calcification. METHODS: The effect of trichostatin A (TSA), an HDI, on human aortic smooth muscle cells (HASMCs) was determined. The mineralization of HASMCs was induced by inorganic phosphorus (Pi), and was confirmed by quantitation of Ca levels and by von Kossa staining. Furthermore, we examined the effect of alkaline phosphatase (ALP) suppression using siRNA on Pi-induced vascular calcification in the presence or absence of TSA. RESULTS: TSA increased the expression and activity of ALP in HASMCs at a concentration which showed an inhibitory effect of histone deacetylase (HDAC) activity but not on cell viability. Moreover, TSA promoted the Pi-induced mineralization of HASMCs. In addition, both phosphonoformic acid (PFA), which is a sodium-dependent phosphate transporter inhibitor, and suppression of ALP expression by siRNA markedly inhibited the TSA-promoted mineralization of HASMCs. CONCLUSION: These data show that inhibition of HDAC activity promotes Pi-induced vascular calcification via the up-regulation of ALP expression. Taken together, HDIs may increase the risk of vascular calcification in CKD patients.

Azechi T; Kanehira D; Kobayashi T; Sudo R; Nishimura A; Sato F; Wachi H

2013-01-01

368

Detection of alkaline phosphatase in canine cells previously stained with Wright-Giemsa and its utility in differentiating osteosarcoma from other mesenchymal tumors.  

UK PubMed Central (United Kingdom)

BACKGROUND: Osteosarcoma (OSA) is a common primary bone tumor in dogs. Demonstration of alkaline phosphatase (ALP) reactivity by tumor cells on unstained slides is useful in differentiating osteosarcoma from other types of sarcoma. However, unstained slides are not always available. OBJECTIVES: The objectives of this study were to evaluate the diagnostic utility of detecting ALP expression in differentiating osteosarcoma from other sarcomas in dogs using cytologic material previously stained with Wright-Giemsa stain and to assess the sensitivity and specificity of ALP expression for diagnosing osteosarcoma using a specific protocol. METHODS: Archived aspirates of histologically confirmed sarcomas in dogs that had been previously stained with Wright-Giemsa stain were treated with 5-bromo, 4-chloro, 3-indolyl phosphate/nitroblue tetrazolium (BCIP/NBT) as a substrate for ALP. Cells were evaluated for expression of ALP after incubation with BCIP/NBT for 1 hour. Sensitivity and specificity of ALP expression for diagnosis of OSA were calculated. RESULTS: In samples from 83 dogs, cells from 15/17 OSAs and from 4/66 tumors other than OSA (amelanotic melanoma, gastrointestinal stromal tumor, collision tumor, and anaplastic sarcoma) expressed ALP. Sensitivity and specificity of ALP expression detected using BCIP/NBT substrate applied to cells previously stained with Wright-Giemsa stain for OSA were 88 and 94%, respectively. CONCLUSIONS: ALP expression detected using BCIP/NBT substrate applied to previously stained cells is useful in differentiating canine OSA from other mesenchymal neoplasms.

Ryseff JK; Bohn AA

2012-09-01

369

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; Shymal Naskar; Binoy K. Mandal

2006-01-01

370

Langmuir films from human placental membranes: preparation, rheology, transfer to alkylated glasses, and sigmoidal kinetics of alkaline phosphatase in the resultant Langmuir-Blodgett film.  

Science.gov (United States)

In the present study, we studied the activity of human placental alkaline phosphatase (PLAP) constraint in a planar surface in controlled molecular packing conditions. For the first time, Langmuir films (LFs) were prepared by the spreading of purified placental membranes (PPM) on the air-water interface and their stability and rheological properties were studied. LFs exhibited a collapse pressure pi(C) = 48 mN/m, hysteresis during the compression-decompression cycle (C-D), indicating a plastic deformation, and a compressibility modulus (K) compatible with liquid-expanded phases. A phase transition point appeared at pi(T) = 28 mN/m and, following successive C-D, it moved toward lower surface areas and higher K, suggesting the lost of some non-PLAP proteins as components of vesicles that might protrude from the monolayer (confirmed by combining lipid/protein molar ratio analysis, PAGE-SDS and V(max)). LFs were transferred at 35 mN/m to alkylated glasses to obtain Langmuir-Blodgett films (LB(35)) the stability of which was confirmed by AFM. The kinetics of p-nitrophenyl phosphate (pNPP) hydrolysis at 37 degrees C catalyzed by PPM was Michaelian and exhibited the thermostability at 60 degrees C typical of PLAP. In LB(35), PLAP exhibited a sigmoidal kinetics which resembled the behavior of the partially metalated enzyme but might become from a cross-talk between protein and membrane structures. PMID:20033626

Clop, Eduardo M; Perillo, María A

2010-04-01

371