WorldWideScience
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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/ < 5 ng/ml). Pearson correlation was applied to test the correlation of serum alkaline phosphatase levels with serum vitamin D/sub 3/ levels. P-value < 0.05 was considered to be significant. Results: Out of 110 samples, 26 had mild (23%), 61 had moderate (55%) and 21 had severe (19.1%) vitamin D deficiencies. All of the patients in the three groups had alkaline phosphatase with in normal limits and the total mean value of the enzyme was 135.97 +- 68.14I U/L. The inter group comparison showed highest values of alkaline phosphatase in the moderate vitamin D deficiency group. The correlation coefficient of alkaline phosphatase and serum vitamin D/sub 3/ levels was r =0.05 (p =0.593). Conclusion: Serus was r =0.05 (p =0.593). Conclusion: Serum vitamin D/sub 3/ levels may not be correlated with increased serum alkaline phosphatase levels. Therefore, alkaline phosphatase may not be used as a screening test to rule out vitamin D deficiency. (author)

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Serum Alkaline Phosphatase Predicts Mortality among Maintenance Hemodialysis Patients  

OpenAIRE

Several observational studies have demonstrated that serum levels of minerals and parathyroid hormone (PTH) have U- or J-shaped associations with mortality in maintenance hemodialysis patients, but the relationship between serum alkaline phosphatase (AlkPhos) and risk for all-cause or cardiovascular death is unknown. In this study, a 3-yr cohort of 73,960 hemodialysis patients in DaVita outpatient dialysis were studied, and the hazard ratios for all-cause and cardiovascular death were higher ...

Regidor, Deborah L.; Kovesdy, Csaba P.; Mehrotra, Rajnish; Rambod, Mehdi; Jing, Jennie; Mcallister, Charles J.; Wyck, David; Kopple, Joel D.; Kalantar-zadeh, Kamyar

2008-01-01

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Peripartal changes in serum alkaline phosphatase activity and lactate dehydrogenase activity in dairy cows.  

OpenAIRE

Peripartal serum alkaline phosphatase activity and lactate dehydrogenase activity were measured in 30 dairy cows in order to examine the association between retained fetal membranes and enzyme activity. Daily blood samples were obtained from pregnant cows, starting 15 days before the expected day of calving until eight days after parturition. Sera from 15 cows which retained fetal membranes longer than 24 hours and 15 cows which shed fetal membranes within six hours after parturition were ana...

Peter, A. T.; Bosu, W. T.; Macwilliams, P.; Gallagher, S.

1987-01-01

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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)

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Relationship between serum heat-stable alkaline phosphatase level and pregnancy  

International Nuclear Information System (INIS)

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

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Association between alendronate, serum alkaline phosphatase level, and heterotopic ossification in individuals with spinal cord injury.  

Science.gov (United States)

Context/objective Only sparse evidence exists regarding the effectiveness of oral alendronate (ALN) in the prevention of heterotopic ossification (HO) in patients with spinal cord injury (SCI). The objective of this study is to investigate the protective effect of oral ALN intake on the appearance of HO in patients with SCI. Study design Retrospective database review. Setting A Spinal Cord Unit at a Rehabilitation Hospital. Participants Two hundred and ninety-nine patients with SCI during acute inpatient rehabilitation. Interventions Administration of oral ALN. Outcome measures The incidence of HO during rehabilitation was compared between patients with SCI receiving oral ALN (n = 125) and patients with SCI not receiving oral ALN (n = 174). The association between HO and/or ALN intake with HO risk factors and biochemical markers of bone metabolism were also explored. Results HO developed in 19 male patients (6.35%), however there was no significant difference in the incidence of HO in patients receiving oral ALN or not. The mean odds ratio of not developing versus developing HO given ALN exposure was 0.8. Significant correlation was found between abnormal serum alkaline phosphatase (ALP) levels and HO appearance (P < 0.001) as well as normal serum ALP and ALN intake (P < 0.05). Conclusion Even though there was no direct prevention of HO in patients with SCI by oral ALN intake, abnormal serum ALP was found more frequently in patients with HO development and without oral ALN intake. This evidence could suggest that ALN may play a role in preventing HO, especially in patients with acute SCI with increasing levels of serum ALP. PMID:24820653

Ploumis, Avraam; Donovan, Jayne M; Olurinde, Mobolaji O; Clark, Dana M; Wu, Jason C; Sohn, Douglas J; O'Connor, Kevin C

2015-03-01

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

International Nuclear Information System (INIS)

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

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

OpenAIRE

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

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

1990-01-01

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

Directory of Open Access Journals (Sweden)

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

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

2012-01-01

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Alkaline phosphatase and cholestasis.  

Science.gov (United States)

The process of cholestasis in both man and rat leads in the majority of cases to the appearance of a biliary band in the electrophoresis of alkaline phosphatase isoenzymes. In this article, the biochemical nature and mode of formation of the biliary band is discussed, with reference to its appearance in cholestasis and other hepatobiliary diseases. PMID:3532919

Wulkan, R W; Leijnse, B

1986-07-01

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Hydrophobic properties of alkaline phosphatases.  

Science.gov (United States)

The butanol extraction method of Morton (1950), a routine step in enzyme purification, is discussed with special reference to a hydrophobic form of alkaline phosphatase from human liver tissue. This form slowly precipitates from butanol-extracted liver tissue homogenates stored at 4 degrees C. Furthermore, it is lost when acetone precipitation is applied as a purification procedure. The soluble form in liver tissue is shown to have a higher relative hydrophobicity than the serum liver/bone isoenzyme. The use of sodium cholate in the isolation of the hydrophobic form produces an artefact in isoelectric focusing, which can be abolished by dialysis prior to focusing. PMID:3803695

Wulkan, R W; Huijskes-Heins, M I; Leijnse, B

1986-01-01

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

International Nuclear Information System (INIS)

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

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Association of alkaline phosphatase phenotypes with arthritides  

Directory of Open Access Journals (Sweden)

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

Padmini A

2004-01-01

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Histomorphometric alteration of knee articular cartilage and serum alkaline phosphatase in young female mice by chronic supplementation with soybean.  

Science.gov (United States)

The purpose of the present study was to examine the effect of soybean supplementation on cartilage thickness in the knee joint and serum levels of alkaline phosphatase (ALP) in mice. Forty female mice were fed for 6 months on one of four regimens: low protein, complete protein without soybean, and complete protein containing either 20% or 40% soybean. Body weight differences, histological and histomorphometric analysis, and ALP levels were determined and compared after 6 months. The results showed a significant increase in serum ALP activity and cartilage thickness in both groups fed on soybean-containing diets, compared with the other groups. Additionally, the number of chondrocytes was significantly increased (p < 0.001) in the group taking the 40% soybean regimen, and the proteoglycan content of the intracellular fluid in the tibia was higher in those groups taking soybean. In conclusion, the present study suggests that soybean supplementation is capable of stimulating ALP production and reducing cartilage loss in young female mice. Soybean supplementation during childhood may therefore be potentially useful in protecting joints. PMID:21110395

Fazelipour, S; Tootian, Z; Matini, E; Hadipour-Jahromy, M

2011-06-01

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Isoelectric focusing of alkaline phosphatases in agarose gel.  

Science.gov (United States)

An isoelectric focusing technique in agarose gel is presented which is suitable for alkaline phosphatases from both serum and tissue sources. An anomaly in the literature about isoelectric focusing of serum alkaline phosphatase from liver origin is discussed and a possible explanation is proposed. The presented technique is used to demonstrate some differences in behaviour of serum liver and bone isoenzymes towards neuraminidase treatment. PMID:4065404

Wulkan, R W; Huijskes-Heins, M I; Leijnse, B

1985-01-01

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

International Nuclear Information System (INIS)

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

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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

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

Scientific Electronic Library Online (English)

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

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

2002-09-01

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

OpenAIRE

Three insecticides (Monitor, Talstar and Benzenehexachloride {BHC}), orally administered at the doses of 0.06, 0.5 and 0.2 mg kg 1 body weight respectively for 21 days affected the body weight and biochemical environment of blood and liver in the rats. Talstar treatment resulted in an increase in the body weight while Monitor and BHC treatment reduced the body weight. After 21 days of the Monitor treatment, activities of acid phosphatases increased (55%) and alkaline phosphates decreased (2%)...

Muhammad Nasim Khan; Tahira Sarwar

2003-01-01

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Serum marker potential of placental alkaline phosphatase-like activity in testicular germ cell tumours evaluated by H17E2 monoclonal antibody assay.  

OpenAIRE

A monoclonal antibody (H17E2) was used in a solid-phase localisation of enzyme activity (ILEA) assay to evaluate placental-like alkaline phosphatase (PLAP) as a serum marker of testicular germ cell tumours. Single or repeated assays were performed on 213 normal blood donor and a smaller number of term pregnancy and testicular cancer sera. The detection limit of PLAP by this system was 0.14 O.D. units equivalent to 0.04iul-1. Of 50 patients with established metastatic disease tested before tre...

Tucker, D. F.; Oliver, R. T.; Travers, P.; Bodmer, W. F.

1985-01-01

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Effect of varying doses of ?-rays on the content of isoenzymes of alkaline phosphatase in the blood serum and small intestine mucosa in rats  

International Nuclear Information System (INIS)

The effect of ?-irradiation with doses ranging from 350 to 15000 rad on the content of the alkaline phosphatase isoenzymes, identified in the blood serum and small intestine mucosa of rats by means of electrophoresis in the agar gel, has been investigated. The most profound lesions have been detected by the 3d day after the exposure to 1000 rad. At the doses above 1000 rad, the decrease in the activity of isoenzymes is less pronounced with the increasing content of isoforms that are detected in the small intestine mucosa of the newborn rats. These facts may be explained by the mechanisms of pathogenic effects of different gamma-ray doses

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21 CFR 864.7660 - Leukocyte alkaline phosphatase test.  

Science.gov (United States)

...2010-04-01 false Leukocyte alkaline phosphatase test. 864.7660 Section 864... § 864.7660 Leukocyte alkaline phosphatase test. (a) Identification. A leukocyte alkaline phosphatase test is a device used to...

2010-04-01

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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)

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 activityiation 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)

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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  

Directory of Open Access Journals (Sweden)

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

Han Ju

2012-02-01

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

DEFF Research Database (Denmark)

Recent studies have demonstrated that growth hormone (GH)-deficient adults have a markedly decreased bone mineral content compared to healthy adults. However, there are conflicting results regarding the effects of GH treatment on bone mineral content in GH-deficient adults. Therefore, we evaluated the effect of GH treatment on a marker of bone formation (bone alkaline phosphatase), hepatic excretory function and distal forearm bone mineral content in GH-deficient adults. Growth hormone was administered subcutaneously in 21 adults (13 males and 8 females) with GH deficiency of childhood onset for 4 months in a double-blind, placebo-controlled GH trial, while 13 of the patients then received further GH for an additional 14 months. Serum insulin-like growth factor I (IGF-I) increased significantly from 100 to 279 micrograms/l and IGF binding protein-3 (IGFBP-3) from 1930 to 3355 micrograms/l after 4 months of GH treatment (p <0.0001). In addition, the molar ratio between IGF-I and IGFBP-3 increased significantlyfrom 0.22 to 0.33 after GH treatment (p <0.0001). Bone alkaline phosphatase increased significantly from 38.6 to 92.9 U/l during GH therapy in male patients (p <0.0001), whereas liver-derived alkaline phosphatase was unaltered by GH. In the females, the increase in bone alkaline phosphatase did not reach statistical significance (19.1 vs 40.0 U/l, p = 0.06). The GH-induced increase in bone alkaline phosphatase correlated significantly with the increase in serum IGFBP-3 (r = 0.46, p = 0.04) but not with the increase in serum IGF-I (p = 0.16).(ABSTRACT TRUNCATED AT 250 WORDS)

Juul, A; Pedersen, S A

1994-01-01

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

OpenAIRE

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

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

1997-01-01

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INEXPENSIVE CHEMIFLUORESCENT DETECTION OF ANTIBODY ALKALINE PHOSPHATASE CONJUGATES ON WESTERN BLOTS USING 4-METHYLUMBELLIFERYL PHOSPHATE.  

Science.gov (United States)

The phosphatase substrate 4-methylumbelliferyl phosphate was tested as a fluorescent reporter for the detection of alkaline phosphatase (AP)-conjugated antibody on immunoblots. Dilution of bovine serum albumin (BSA) was used as a protein target with the use of anti-BSA antisera as the primary react...

28

Human placental alkaline phosphatase in liver and intestine  

International Nuclear Information System (INIS)

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

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21 CFR 862.1050 - Alkaline phosphatase or isoenzymes test system.  

Science.gov (United States)

...2010-04-01 false Alkaline phosphatase or isoenzymes test system. 862...Systems § 862.1050 Alkaline phosphatase or isoenzymes test system. (a) Identification. An alkaline phosphatase or isoenzymes test system is...

2010-04-01

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Phosphorylation by Alkaline Phosphatase: Immobilization and Synthetic Potential  

Directory of Open Access Journals (Sweden)

Full Text Available Phosphatases (AP, E.C. 3.1.3.1 are hydrolytic enzymes that naturally hydrolyse phosphomonoesters but in a so-called transphosphorylation reaction these enzymes are also able to transfer a phosphate group from phosphorylated compounds to alcoholic functions. This transphosphorylation catalysed by acid phosphatases using pyrophosphate as a phosphate donor has been studied in some detail. However, the acidic pH optimum of these enzymes limits some of their applications. The catalytic features of alkaline phosphatase are similar to the acid phosphatases and its alkaline pH optimum suggests a possible application of this enzyme in phosphorylation reactions which need to be carried out at higher pH. Here we explore the synthetic potential of bovine intestine alkaline phosphatase (AP in the phosphorylation of dihydroxyacetone (DHA and glycerol using pyrophosphate (PPi as phosphate donor. The phosphorylated compounds are intermediates in two multi-enzymatic cascade reactions for the synthesis of carbohydrates. The yields of dihydroxyacetone phosphate (DHAP and glycerol-1-phosphate at pH 8 (2.6 mM and 2.2 mM, respectively were comparable to the results obtained with the acid phosphatases at pH 4. Nevertheless, when the cascade reactions were carried out at pH 8, very low conversions were measured due to inactivation of the alkaline phosphatase by the product phosphate. To circumvent this inhibition, the alkaline phosphatase was immobilized on aldehyde-activated beads (Sepabeads EC-HA. The immobilization greatly diminished the inhibition by phosphate, and the immobilized alkaline phosphatase at pH 8 gave the same conversions in the cascade reaction starting from DHA as obtained with the acid phosphatase at pH 6. However, the immobilized enzyme was active for only one catalytic cycle and the beads could not be reused.

Lara Babich

2013-05-01

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

International Nuclear Information System (INIS)

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

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Overexpression of Human Bone Alkaline Phosphatase in Pichia Pastoris  

Science.gov (United States)

The Pichiapastoris expression system was utilized to produce functionally active human bone alkaline phosphatase in gram quantities. Bone alkaline phosphatase is a key enzyme in bone formation and biomineralization, yet important questions about its structural chemistry and interactions with other cellular enzymes in mineralizing tissues remain unanswered. A soluble form of human bone alkaline phosphatase was constructed by deletion of the 25 amino acid hydrophobic C-terminal region of the encoding cDNA and inserted into the X-33 Pichiapastoris strain. An overexpression system was developed in shake flasks and converted to large-scale fermentation. Alkaline phosphatase was secreted into the medium to a level of 32mgAL when cultured in shake flasks. Enzyme activity was 12U/mg measured by a spectrophotometric assay. Fermentation yielded 880mgAL with enzymatic activity of 968U/mg. Gel electrophoresis analysis indicates that greater than 50% of the total protein in the fermentation is alkaline phosphatase. A purification scheme has been developed using ammonium sulfate precipitation followed by hydrophobic interaction chromatography. We are currently screening crystallization conditions of the purified recombinant protein for subsequent X-ray diffraction analyses. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

Karr, Laurel; Malone, Christine, C.; Rose, M. Franklin (Technical Monitor)

2000-01-01

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Advanced stage of breast cancer hoist alkaline phosphatase activity: risk factor for females in India  

OpenAIRE

Breast cancer is the most common neoplasm affecting women in the western world with an average frequency of 1 in 11, developing the malignancy and it is second most common cancer in India. Variations in serum levels of biochemical parameters especially alkaline phosphatase (ALP) changes may be of great help in diagnosis of breast carcinoma. Serum ALP activity was assayed in 388 histopathologically proven breast cancer patients using spectrophotometric methods and monitored association with ca...

Singh, A. K.; Pandey, A.; Tewari, M.; Kumar, R.; Sharma, A.; Singh, K. A.; Pandey, H. P.; Shukla, H. S.

2013-01-01

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

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

Fatemeh Afshari

2013-07-01

35

Characterization of Human Bone Alkaline Phosphatase in Pichia Pastoris  

Science.gov (United States)

A soluble form of human bone alkaline phosphatase has been expressed in a recombinant strain of the methylotrophic yeast Pichia pastoris. We constructed a plasmid containing cDNA encoding for human bone alkaline phosphatase, with the hydrophobic carboxyl terminal portion deleted. Alkaline phosphatase was secreted into the medium to a level of 32mg/L when cultured in shake flasks, and enzyme activity was 12U/mg, as measured by a spectrophotometric assay. By conversion to a fermentation system, a yield of 880mg/L has been achieved with an enzyme activity of 968U/mg. By gel electrophoresis analysis, it appears that greater than 50% of the total protein in the fermentation media is alkaline phosphatase. Although purification procedures are not yet completely optimized, they are expected to include filtration, ion exchange and affinity chromatography. Our presentation will focus on the purification and crystallization results up to the time of the conference. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

Malone, Christine C.; Ciszak, Eva; Karr, Laurel J.

1999-01-01

36

Sensitive optical detection of alkaline phosphatase activity with quantum dots  

Energy Technology Data Exchange (ETDEWEB)

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

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

2014-01-15

37

Sensitive optical detection of alkaline phosphatase activity with quantum dots  

International Nuclear Information System (INIS)

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

38

A highly active alkaline phosphatase from the marine bacterium cobetia.  

Science.gov (United States)

An alkaline phosphatase with unusually high specific activity has been found to be produced by the marine bacterium Cobetia marina (strain KMM MC-296) isolated from coelomic liquid of the mussel Crenomytilus grayanus. The properties of enzyme, such as a very high specific activity (15000 DE U/1 mg of protein), no activation with divalent cations, resistance to high concentrations of inorganic phosphorus, as well as substrate specificity toward 5' nucleotides suggest that the enzyme falls in an intermediate position between unspecific alkaline phosphatases (EC 3.1.3.1) and 5' nucleotidases (EC 3.1.3.5). PMID:15906116

Yu Plisova, E; Balabanova, L A; Ivanova, E P; Kozhemyako, V B; Mikhailov, V V; Agafonova, E V; Rasskazov, V A

2005-01-01

39

Human placental alkaline phosphatase electrophoretic alleles: Quantitative studies  

OpenAIRE

Human placental alkaline phosphatase (ALP) activity has been determined in specimens obtained from 562 Italian subjects. The mean activities of the three common homozygotes (Pl 2 = 4.70 ± 0.24, Pl 1 = 4.09 ± 0.08, and Pl 3 = 2.15 ± 0.71 ?mol of p-nitrophenol produced) were significantly different.

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

1982-01-01

40

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

2006-03-01

41

Microvillus inclusion disease: specific diagnostic features shown by alkaline phosphatase histochemistry.  

OpenAIRE

A technique using alkaline phosphatase histochemistry on routine sections of four jejunal biopsy specimens and one necropsy sample was applied to show that alkaline phosphatase activity, normally present in the brush border, occurs in the enterocytes of patients with microvillus inclusion disease. Sections were cut at 5 micron, mounted on to glass slides, and dried overnight at 37 degrees C before staining for alkaline phosphatase activity by the indoxyl phosphatase nitro blue tetrazolium met...

Lake, B. D.

1988-01-01

42

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

OpenAIRE

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

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

1983-01-01

43

Intestinal alkaline phosphatase inhibits the proinflammatory nucleotide uridine diphosphate  

OpenAIRE

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....

Moss, Angela K.; Hamarneh, Sulaiman R.; Mohamed, Mussa M. Rafat; Ramasamy, Sundaram; Yammine, Halim; Patel, Palak; Kaliannan, Kanakaraju; Alam, Sayeda N.; Muhammad, Nur; Moaven, Omeed; Teshager, Abeba; Malo, Nondita S.; Narisawa, Sonoko; Milla?n, Jose? Luis; Warren, H. Shaw

2013-01-01

44

Alkaline Phosphatase from Venom of the Endoparasitoid Wasp, Pteromalus puparum  

OpenAIRE

Using chromogenic substrates 5-bromo-4-chloro-3?-indolyl phosphate and nitro blue tetrazolium, alkaline phosphatase (ALPase) was histochemically detected in the venom apparatus of an endoparasitoid wasp, Pteromalus puparum L. (Hymenoptera: Pteromalidae). Ultrastructural observations demonstrated its presence in the secretory vesicles and nuclei of the venom gland secretory cells. Using p-nitrophenyl phosphate as substrate to measure enzyme activity, the venom ALPase was found to be temperat...

Zhu, Jia-ying; Yin Ye, Gong; Fang, Qi; Hu, Cui

2010-01-01

45

An alkaline phosphatase reporter for use in Clostridium difficile.  

Science.gov (United States)

Clostridium difficile is an anaerobic, Gram-positive pathogen that causes severe gastrointestinal disease in humans and other mammals. C. difficile is notoriously difficult to work with and, until recently, few tools were available for genetic manipulation and molecular analyses. Despite the recent advances in the field, there is no simple or cost-effective technique for measuring gene transcription in C. difficile other than direct transcriptional analyses (e.g., quantitative real-time PCR and RNA-seq), which are time-consuming, expensive and difficult to scale-up. We describe the development of an in vivo reporter assay that can provide qualitative and quantitative measurements of C. difficile gene expression. Using the Enterococcus faecalis alkaline phosphatase gene, phoZ, we measured expression of C. difficile genes using a colorimetric alkaline phosphatase assay. We show that inducible alkaline phosphatase activity correlates directly with native gene expression. The ability to analyze gene expression using a standard reporter is an important and critically needed tool to study gene regulation and design genetic screens for C. difficile and other anaerobic clostridia. PMID:25576237

Edwards, Adrianne N; Pascual, Ricardo A; Childress, Kevin O; Nawrocki, Kathryn L; Woods, Emily C; McBride, Shonna M

2015-04-01

46

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

Science.gov (United States)

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

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

2013-10-01

47

ALKALINE PHOSPHATASE ACTIVITY AS A MARKER OF DOG SEMEN FREEZABILITY  

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

KOSINIAK-KAMYSZ K.

2007-01-01

48

Initial activity and inactivation of alkaline phosphatase in different lots of buffer.  

Science.gov (United States)

Alkaline phosphatase activities were determined in six lots of 2-amino-2-methyl-1-propanol (AMP) and in six lots of diethanolamine (DEA) buffers without preincubation of the sample. There appeared to be differences between the lot numbers in both cases, resulting in a variation in initial activity. When serum samples are preincubated with buffer a loss of activity was observed in 4 out of the 6 AMP buffers. Four human isoenzymes showed varying inactivation during preincubation with AMP buffer. No loss of activity was observed when the preincubation was done with the six DEA buffers. These results indicate that the purity of the commercially-available buffers is quite unsatisfactory. PMID:657529

Pekelharing, J M; Leijnse, B

1978-05-01

49

[Effect of polyenic antibiotics on the activity of alkaline phosphatase from Candida albicans].  

Science.gov (United States)

Polyenic antibiotics (levorin, amphotericin B, nistatin) inhibit in vivo and in vitro the activity of membrane alkaline phosphatase from sensitive Candida albicans strain, and their inhibitory effect is twice lower on the enzyme from the resistant strain. A correlation is observed between the antibiotic concentration and the inhibitory effect on alkaline phosphatase activity. Nistatin is found to be the least efficient inhibitor (among the antibiotics studied) of alkaline phosphatase. The treatment of membranes with polyenic antibiotics does not result in solubilization of membrane proteins nad alkaline phosphatase. The data obtained are considered with respect to the effect of polyenic antibiotics on cell membrane structure. PMID:322734

Solov'ena, N N; Belousova, I I; Tereshin, I M

1977-02-01

50

Improved double immunohistochemical staining method for cryostat and paraffin wax sections, combining alkaline phosphatase anti-alkaline phosphatase and indirect immunofluorescence.  

OpenAIRE

Aims - To develop an immunohistochemical staining method for cryostat and paraffin wax sections so that two different antigens in the same section of tissues could be detected by combining immunoenzyme and immunofluorescence techniques. Methods - This double immunohistochemical staining method combines alkaline phosphatase-anti-alkaline phosphatase (APAAP) using New Fuchsin as a chromogen and indirect immunofluorescence. Results - APAAP staining for one antigen of this double immunohistochemi...

Tao, Q.; Srivastava, G.; Loke, S. L.; Chan, E. Y.; Ho, F. C.

1994-01-01

51

The influence of complexing pharmaceutical compositions on alkaline phosphatase  

Science.gov (United States)

It is established that the pharmaceutical compositions xydiphon, medifon, succimer, and EDTA, which are used as complexing agents for accelerating the excretion of heavy metals from human organism, at certain concentrations inhibit enzyme alkaline phosphatase (AP). It is concluded that xydiphon and EDTA have a noticeable effect on AP activity at concentrations over 0.01 mM; medifon and succimer, at concentrations of over 0.3-0.5 mM. The enzyme's inhibition constants and type of inhibition are determined. Xydiphon is found to manifest the highest affinity to AP ( K I = 0.35 mM). It is shown by kinetic analysis that dissociative chemoinactivation of the enzyme takes place under the action of complexing agents. The corresponding kinetic parameters are calculated.

Atyaksheva, L. F.; Chukhrai, E. S.; Stepina, N. D.; Novikova, N. N.; Yur'eva, E. A.

2011-06-01

52

New antibody in severe rhesus incompatible pregnancies: IgG-kappa antiplacental alkaline phosphatase.  

Science.gov (United States)

As already found in other various diseases, a macromolecular alkaline phosphatase complex (HMW-AP) was also found in sera of two severe Rhesus-incompatible pregnancies complicated by ascites and fetal hydrops at delivery. This atypical complex was detected and isolated by agarose gel electrophoresis. Immunoelectrophoresis and heat inactivation of this HMW-AP complex revealed that it consisted of IgG of the kappa type and placental AP isoenzyme. The transitory presence of this immuncomplex is discussed. However, in all women with Rh-immunized complicated pregnancies, significant variations of neutrophil and serum AP activities were observed. A fall in AP activity and the presence of an antiplacental AP antibody in serum of women with complicated Rh immunization should be of value in assessing the prognosis of the disease. PMID:2504185

Brisson-Lougarre, A; Vergnes, H; Grozdea, J; Alie-Daram, S; Fontanilles, A M; Bierme, R

1989-04-01

53

Histochemical imaging of alkaline phosphatase using a novel fluorescent substrate.  

Science.gov (United States)

Histochemical visualization of phosphatase is exclusively required for Western immunoblotting and antigen-positive cell staining using an alkaline phosphatase (AP)-labeled secondary antibody. This detection has been performed by several reagents including 5-bromo-4-chloro-3-indolyl-phosphate (X-Phos), nitro blue tetrazolium (NBT), 3-(2'-spiroadamantane)-4-methoxy-4-(3?-phosphoryloxy)phenyl-1,2-dioxetane and 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-[3H]-quinazolinone (ELF® 97 Phosphate). We previously reported that 2-(benzothiazol-2-yl)-4-bromophenol bonded with N-acetylneuraminic acid (BTP3-Neu5Ac), enabled fluorescent histochemical visualization of sialidase activity. 2-(Benzothiazol-2-yl)-4-bromophenol (BTP3), which is formed from BTP3-Neu5Ac by sialidase reaction, is a crystalline, insoluble and stable fluorogenic compound, deposited at the site of enzyme activity. We developed a BTP3 phosphate ester (BTP3-Phos) for the purpose of fluorescent histochemical visualization of phosphatase activity. BTP3-Phos emitted fluorescence in a manner dependent on the concentration of the AP-labeled antibody. BTP3-Phos also enabled fluorescent histochemical visualization of AP-blotted dots in a manner dependent on the concentration of the AP-labeled antibody. The detection sensitivity of BTP3-Phos was estimated to be greater than that of the conventional method using X-Phos and NBT. Influenza A virus-infected cells were fixed and reacted with anti-influenza A virus antibodies and incubated continuously with an AP-labeled secondary antibody. BTP3-Phos stained the infected cells with distinct green fluorescence. These results indicate that BTP3-Phos can enable fluorescent immunohistochemical staining analysis using an AP-labeled antibody. BTP3-Phos would be beneficial for histochemical staining of AP activity, and may be applicable for multi-color staining or a cell sorter. PMID:25109307

Takahashi, Tadanobu; Otsubo, Tadamune; Ikeda, Kiyoshi; Minami, Akira; Suzuki, Takashi

2014-01-01

54

Microvillus inclusion disease: specific diagnostic features shown by alkaline phosphatase histochemistry.  

Science.gov (United States)

A technique using alkaline phosphatase histochemistry on routine sections of four jejunal biopsy specimens and one necropsy sample was applied to show that alkaline phosphatase activity, normally present in the brush border, occurs in the enterocytes of patients with microvillus inclusion disease. Sections were cut at 5 micron, mounted on to glass slides, and dried overnight at 37 degrees C before staining for alkaline phosphatase activity by the indoxyl phosphatase nitro blue tetrazolium method. Incubation periods amounted to 10 minutes for biopsy specimens and 30 minutes to one hour for necropsy samples. The demonstration of alkaline phosphatase activity in routinely processed biopsy specimens provides an effective, quick, and definitive test in the diagnosis of microvillus inclusion disease without recourse to electron microscopy. PMID:3170775

Lake, B D

1988-08-01

55

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

Science.gov (United States)

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

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

2009-08-01

56

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

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

Ferda CAGAVI

2005-06-01

57

Induction of glomerular alkaline phosphatase after challenge with lipopolysaccharide  

Science.gov (United States)

Alkaline phosphatase (AP) can be considered as a host defence molecule since this enzyme is able to detoxify bacterial endotoxin at physiological pH. The question emerged whether this anti-endotoxin principle is inducible in the glomerulus and if so, which glomerular cells might be involved in the expression of ectoAP after stimulation with pro-inflammatory agents. Therefore kidneys of rats treated with either lipopolysaccharide (LPS), E. coli bacteria or non-toxic monophosphoryl lipid A (MPLA) were examined for AP activity 6 or 24 h after challenge. In addition cultures of endothelial cells or mesangial cells were evaluated for AP activity after stimulation with either LPS, TNF? or IL-6, and mRNA for AP was studied in TNF?-stimulated and control mesangial cells. The results show significant up-regulation of glomerular AP in LPS- or E. coli-injected rats compared to rats injected with MPLA. Endothelial and mesangial cells in vitro showed significant up-regulation of AP activity following stimulation with LPS, TNF? or IL-6, whereas increased mRNA for AP was observed in mesangial cells after TNF? stimulation compared to non-stimulated control cells. Since it appeared that hydrolysis occurred when endotoxin was used as a substrate in the histochemical staining, we concluded that inducible glomerular ectoAP may reflect a local endotoxin detoxifying principle of the kidney. PMID:12974943

Kapojos, Jola J; Poelstra, Klaas; Borghuis, Theo; Van Den Berg, Anke; Baelde, Hans J; Klok, Pieter A; Bakker, Winston W

2003-01-01

58

How Should an Increase in Alkaline Phosphatase Activity Be Interpreted?  

International Nuclear Information System (INIS)

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

59

Alkaline phosphatase from venom of the endoparasitoid wasp, Pteromalus puparum.  

Science.gov (United States)

Using chromogenic substrates 5-bromo-4-chloro-3'-indolyl phosphate and nitro blue tetrazolium, alkaline phosphatase (ALPase) was histochemically detected in the venom apparatus of an endoparasitoid wasp, Pteromalus puparum L. (Hymenoptera: Pteromalidae). Ultrastructural observations demonstrated its presence in the secretory vesicles and nuclei of the venom gland secretory cells. Using p-nitrophenyl phosphate as substrate to measure enzyme activity, the venom ALPase was found to be temperature dependent with bivalent cation effects. The full-length cDNA sequence of ALPase was amplified from the cDNA library of the venom apparatus of P. puparum, providing the first molecular characterization of ALPase in the venom of a parasitoid wasp. The cDNA consisted of 2645 bp with a 1623 bp open reading frame coding for 541 deduced amino acids with a predicted molecular mass of 59.83 kDa and pI of 6.98. Using multiple sequence alignment, the deduced amino acid sequence shared high identity to its counterparts from other insects. A signal peptide and a long conserved ALPase gene family signature sequence were observed. The amino acid sequence of this venom protein was characterized with different potential glycosylation, myristoylation, phosphorylation sites and metal ligand sites. The transcript of the ALPase gene was detected by RT-PCR in the venom apparatus with development related expression after adult wasp emergence, suggesting a possible correlation with the oviposition process. PMID:20575745

Zhu, Jia-Ying; Yin Ye, Gong; Fang, Qi; Hu, Cui

2010-01-01

60

Activity and isoenzyme spectrum of alkaline phosphatase in liver and blood plasma of gamma-irradiated hen and chick embryos  

International Nuclear Information System (INIS)

Twelve day and 20 day-old embryos and one-day old chicks were gamma-irradiated with a single dose of 100 rad. On the 1st, 2nd and 72nd hours after treatment the alkaline phosphatase (AP) activity and isozyme spectrum in liver homogenates and blood plasma were determined. AP in the liver was demonstrated histochemically as well. The results show that the initial damage of the liver parenchyma by irradiation treatment is characterized by a slight decrease of liver AP activity and a marked increase in total plasma enzyme activity. The isozyme spectrum changes (increase of AP1 in the early period and increase of AP1 and AP5 in the later period) show that the initial liver parenchymal damage is followed by bile duct damage as well. Comparison of the results of biochemical and histochemical studies indicate the presence of direct correlation between serum and liver alkaline phosphatase activities. (A.B.)

61

Cloning and expression of a highly active recombinant alkaline phosphatase from psychrotrophic Cobetia marina.  

Science.gov (United States)

Alkaline phosphatase catalyzes the hydrolysis of phosphomonoesters and is widely used in molecular biology techniques and clinical diagnostics. We expressed a recombinant alkaline phosphatase of the marine bacterium, Cobetia marina, in Escherichia coli BL21 (DE3). The recombinant protein was purified with a specific activity of 12,700 U/mg protein, which is the highest activity reported of any bacterial alkaline phosphatase studied to date. The molecular mass of the recombinant protein was 55-60 kDa, as determined by SDS-PAGE, and was observed to be a dimer by gel filtration analysis. The enzyme was optimally active at 45°C and the recombinant alkaline phosphatase efficiently hydrolyzed a phosphoric acid ester in luminescent and fluorescent substrates. Therefore, this enzyme can be considered to be extremely useful as a label conjugated to an antibody. PMID:22009571

Nasu, Eriko; Ichiyanagi, Atsushi; Gomi, Keiko

2012-02-01

62

Value of plasma calcium, phosphate, and alkaline phosphatase measurements in the diagnosis of histological osteomalacia  

OpenAIRE

Plasma calcium and phosphate concentrations and alkaline phosphatase activities were examined retrospectively in 50 patients with histologically proven osteomalacia and 50 age- and sex-matched control subjects with normal bone histology. An abnormal plasma alkaline phosphatase activity was more useful than an abnormal plasma calcium or phosphate concentration in distinguishing between normal and osteomalacic subjects, producing a false-negative rate of 14% and a false-positive rate of 8%. Fal...

Peach, Hedley; Compston, Juliet E.; Vedi, Shobhana; Horton, Leo Wl

1982-01-01

63

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

International Nuclear Information System (INIS)

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

64

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

Energy Technology Data Exchange (ETDEWEB)

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

Ishibe, M.; Rosier, R.N.; Puzas, J.E. (Department of Orthopaedics, University of Rochester, New York (United States))

1991-10-01

65

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

Science.gov (United States)

Human prostatic acid phosphatase (hPAP) directly enhances the differentiated characteristics of isolated bone cells in vitro. This enzyme, when added to cell cultures for 24 h in vitro stimulates collagen synthesis and the production of alkaline phosphatase. The effects are dose dependent, with statistically significant effects occurring from 0.1-100 nM hPAP. Concentrations higher than 100 nM do not evoke greater effects. The maximal effect of hPAP occurs between 12 and 24 h of exposure. The cells stimulated to the greatest degree are osteoprogenitor cells and osteoblasts. Fibroblasts isolated from the same tissue show a lesser sensitivity to hPAP. hPAP has no detectable effect on cell proliferation, as measured by radiolabeled thymidine incorporation or total DNA synthesis. None of the observations reported in this work can be attributed to contaminating proteins in the hPAP preparation. hPAP was radiolabeled with 125I and was used for affinity binding and cross-linking studies. Scatchard analysis of specific binding indicated the presence of 1.0 X 10(5) high affinity binding sites/cell, with a Kd of 6.5 nM. Cross-linking studies demonstrated the presence of one 320-kDa binding complex. The pH profile and kinetic determinations of Km and maximum velocity for hPAP were similar to those previously reported, except for the finding of positive cooperativity of the substrate with the enzyme under the conditions of our assay. We believe that the direct stimulation of bone-forming cells by hPAP may contribute to the sclerotic nature of skeletal bone around sites of neoplastic prostatic metastases and that the effect of the enzyme is probably mediated by a plasma membrane receptor. PMID:1653783

Ishibe, M; Rosier, R N; Puzas, J E

1991-10-01

66

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

2013-07-21

67

Radioimmunoassay for prostatic acid phosphatase in human serum. Methodologic aspects  

International Nuclear Information System (INIS)

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

68

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

International Nuclear Information System (INIS)

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

69

Expression of a human placental alkaline phosphatase gene in transfected cells: use as a reporter for studies of gene expression.  

OpenAIRE

The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is...

Henthorn, P.; Zervos, P.; Raducha, M.; Harris, H.; Kadesch, T.

1988-01-01

70

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

Directory of Open Access Journals (Sweden)

Full Text Available

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

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

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

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

Mauren Picada Emanuelli

2008-04-01

71

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

International Nuclear Information System (INIS)

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

72

Extracellular Ca2(+)-dependent inducible alkaline phosphatase from extremely halophilic archaebacterium Haloarcula marismortui.  

Science.gov (United States)

When starved of inorganic phosphate, the extremely halophilic archaebacterium Haloarcula marismortui produces the enzyme alkaline phosphatase and secretes it to the medium. This inducible extracellular enzyme is a glycoprotein whose subunit molecular mass is 160 kDa, as estimated by sodium dodecyl sulfate-gel electrophoresis. The native form of the enzyme is heterogeneous and composed of multiple oligomeric forms. The enzymatic activity of the halophilic alkaline phosphatase is maximal at pH 8.5, and the enzyme is inhibited by phosphate. Unlike most alkaline phosphatases, the halobacterial enzyme requires Ca2+ and not Zn2+ ions for its activity. Both calcium ions (in the millimolar range) and NaCl (in the molar range) are required for the stability of the enzyme. PMID:2123861

Goldman, S; Hecht, K; Eisenberg, H; Mevarech, M

1990-12-01

73

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

Scientific Electronic Library Online (English)

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

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.

1187-11-01

74

Alpha-1-fetoprotein and the heat stable alkaline phosphatase in some liver diseases.  

Science.gov (United States)

The presence of alpha-1-fetoprotein, the heat stable alkaline phosphatase and Australia antigen was examined in 103 patients with porphyria cutanea tarda, 300 patients with cirrhosis and 18 patients with primary liver carcinoma. The heat stable alkaline phosphatase was determined in 46 percent of patients with porphyria cutanea tarda and in 61 percent of patients with primary liver carcinoma. Alpha-1-fetoprotein was detected in 61 percent of patients with primary liver carcinoma and in 2 patients with porphyria cutanea tarda in whom primary liver carcinoma was proved later. The simultaneous occurrence of alpha-1-fetoprotein and the heat stable alkaline phosphatase was found in 50 percent of cases with primary liver carcinoma. Neither the patients with porphyria cutanea tarda nor the patients with cirrhosis were Australia-antigen positive. Australia-antigen could be detected only in one patient with alpha-1-fetoprotein positive-carcinoma of the liver. PMID:48325

Zizkovský, V; Kordac, V; St?pán, J; El'gort, D N

1975-04-01

75

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

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

Sh Byranvand

2006-10-01

76

Identification of a macro-alkaline phosphatase complex in a patient with inflammatory bowel disease.  

Science.gov (United States)

We report the rare finding of a macro-alkaline phosphatase (macroALP) complex in a patient with a previously unexplained raised alkaline phosphatase activity. The clinical symptoms were persistent, daily diarrhoea for two months with blood in the stool. The patient was subsequently diagnosed with inflammatory bowel disease, specifically ulcerative colitis, following a rectal biopsy and colonoscopy. Two cases of macroALP associated with ulcerative colitis have been reported before, suggesting there could be an increased prevalence of macroALP in these patients. PMID:22454544

McTaggart, Malcolm P; Rawson, Catherine; Lawrence, David; Raney, Barbara S; Jaundrill, Linnet; Miller, Lorna A; Murtinho-Braga, Joseph; Kearney, Edward M

2012-07-01

77

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

OpenAIRE

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 or in the defined medium without cobalt. Stimulation of enzyme activity with cobalt is dependent on a low phosphate concentration in the medium (below 0.075 mM) and continued protein synthesis. Cob...

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

1981-01-01

78

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

OpenAIRE

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

Deepak Ganjewala

2010-01-01

79

Digestive enzyme and alkaline phosphatase activities during the early stages of Silurus soldatovi development.  

Science.gov (United States)

To provide a theoretical basis to improve the survival and growth rate and optimize diet of sheatfish (Silurus soldatovi), the activities of certain digestive enzymes and alkaline phosphatases were investigated during larval development of one-ten day old individuals. Results indicated that sheatfish larva (~ three days after hatching) had high levels of alkaline protease activity, which peaked at five days old and dipped by eight days old, although the trend was generally upward. Acid protease activity at one-eight days old was low, after which it increased rapidly. Amylase activity reached the highest value at five days old, after which it began to decline. Lipase activity fluctuated markedly and showed two peaks at three-four days old and six-eight days old. Larval digestive enzyme activity and alkaline phosphatase activity were higher when fed live food than when fed an artificial diet. Throughout the early development process, alkaline protease activity was higher than acid protease, alkaline protease and amylase specific activity decreased significantly for eight-day-old transition larvae, while acid protease activity increased rapidly. These results indicate that the changes in digestive enzyme activity were relevant to digestive function conversion during fish larvae development. Alkaline phosphatase activity showed an upward trend over the first ten days of life, which indicated that the gastrointestinal function of sheatfish improved gradually. PMID:21174353

Liu, Wei; Zhang, Xiu-Mei; Wang, Li-Bo

2010-12-01

80

Adriamycin alters the alkaline phosphatase activity in hamster molars during development in vitro.  

Science.gov (United States)

The effect of a 2 hour exposure to adriamycin (1 mg/litre) on alkaline phosphatase (ALPase) activity of the golden hamster 4-5 day old second maxillary molars (M2) was investigated in vitro. The molars were grown in BGJb medium containing 15% fetal bovine serum, glutamine (200 micrograms/ml), vitamin C (250 micrograms/ml), penicillin G (50 micrograms/ml), and streptomycin sulphate (30 micrograms/ml). The gas phase contained 50% O2 + 5% CO2 + 45% N2. The molars were supported on cellulosic membrane filters and grown for 3, 5, and 7 days at the medium-gas interface in a closed humidified chamber. Biochemical analysis indicated a steady increase in ALPase activity throughout this study in the control samples. However, after adriamycin treatment no increase in ALPase activity could be observed. The histochemical data showed that the increased activity in the control was confined to the peripheral pulp, sub-odontoblastic layer, stratum intermedium, ameloblasts and odontoblasts. Although these layers showed a decreased activity after adriamycin treatment, the ameloblasts showed an increase in activity over the control. The data has shown that adriamycin caused a reduction in total ALPase activity in developing molars in vitro; osteodentin production by pulp cells; and appeared to produce an acceleration in the differentiation of ameloblasts. PMID:8329861

Karim, A C; Wöltgens, J H; Bervoets, T J; Lyaruu, D M; Bronchers, A L

1993-05-01

81

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

DEFF Research Database (Denmark)

Free fatty acids released during intralumenal digestion of dietary fat must pass through the enterocyte brush border membrane before triacylglycerol reassembly and subsequent chylomicron delivery to the lymph system. In the present work fluorescent BODIPY fatty acid analogs were used to study this membrane passage in organ cultured intestinal mucosal explants. We found that in addition to a rapid uptake into the cytoplasm, a fraction of the fatty acid analogs were inserted directly into the brush border membrane. Furthermore, a brief exposure of microvillar membrane vesicles to a fat mixture mimicking a physiological solution of dietary mixed micelles, rearranged the lipid raft microdomain organization of the membranes. Thus, the fat mixture generated a low-density subpopulation of microvillar detergent resistant membranes (DRMs) highly enriched in alkaline phosphatase (AP). Since this GPI-linked enzyme is the membrane protein in the brush border with the highest affinity for lipid rafts, this implies that free fatty acids selectively insert stably into these membrane microdomains. We have previously shown that absorption of dietary lipids transiently induce a selective endocytosis of AP from the brush border, and from work by others it is known that fat absorption is accompanied by a rise in serum AP and secretion of surfactant-like particles from enterocytes. We propose that these physiological processes may be triggered by the sequestering of dietary free fatty acids in lipid raft microdomains of the brush border.

Hansen, Gert H; Rasmussen, Karina

2011-01-01

82

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

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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

2001-01-01

83

Nature of immobilization surface affects antibody specificity to placental alkaline phosphatase.  

Science.gov (United States)

Retention of native conformation of immobilized protein is essential for various applications including selection and detection of specific recombinant antibodies (scFvs). Placental alkaline phosphatase (PAP), an onco-fetal antigen expressed on the surface of several tumors, was immobilized on supermagnetic particles for selection of recombinant antibodies from a human phage display antibody library. The isolated antibodies were found to be cross-reactive to either of the isozymes of alkaline phosphatase, i.e., bone alkaline phosphatase (BAP) or intestinal alkaline phosphatase (IAP) and could not be used for tumor targeting. A specific anti-PAP monoclonal antibody H17E2 was tested for retention of specificity under these conditions. Binding of the antibody to magnetic beads conjugated IAP and BAP along with PAP and the ability of the two isozymes to inhibit its binding to PAP depicted the loss of isozyme specificity of the antibody. However, the antibody retained its specificity to PAP immobilized on polyvinyl chloride (PVC) surface. Enzyme activity was observed on both surfaces. This demonstrates that nature of immobilization may affect antigen-antibody binding in subtle ways, resulting in alteration of conformation of the epitopes. This may have consequences for determining the specificity of antibody binding for proteins that share a high degree of homology. PMID:25321174

Kumar, Mukesh; Khan, Imran; Sinha, Subrata

2015-01-01

84

Radioprotective effect of MPG and WR-2721 against gamma-radiolysis of human placental alkaline phosphatase  

International Nuclear Information System (INIS)

Two-radioprotective drugs - MPG and WR-2721 have been found to protect human placental alkaline phosphatase against gamma radiolysis. Based on current literature and results obtained here free radical scavenging shielding of active sites and/or conformational change due to binding of the drug may be suggested as the possible mechanism of chemical radioprotection. (author)

85

A dimerization defect caused by a glycine substitution at position 420 by serine in tissue-nonspecific alkaline phosphatase associated with perinatal hypophosphatasia.  

Science.gov (United States)

Mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene cause hypophosphatasia (HPP), an inborn error of metabolism characterized by defects in bone and teeth mineralization accompanying subnormal levels of serum alkaline phosphatase activity. Missense mutations at position 420 of TNSALP (standard nomenclature), which convert glycine to serine [TNSALP (G420S)] or alanine [TNSALP (G420A)], have been reported in perinatal and childhood HPP, respectively. When expressed in COS-1 cells, both TNSALP mutants were indistinguishable from wild-type TNSALP [TNSALP (W)] as evidenced by immunofluorescence and western blotting. Nevertheless, the two TNSALP mutants did not show substantial alkaline phosphatase activity. In agreement with transiently transfected cells, TNSALP (G420S) expressed in a Tet-On inducible expression system lacked its alkaline phosphatase activity, although this mutant was anchored to the cell surface lipid bilayers by glycosylphosphatidylinositol as an 80 kDa mature form bearing complex-type oligosaccharides like TNSALP (W). Importantly, TNSALP (G420S) was found to largely fail to assemble into the homodimer in contrast to TNSALP (W). Taken together, these results demonstrate that the glycine residue at position 420 is crucial for the subunit interaction of TNSALP and hence its catalytic function without affecting trafficking of monomeric TNSALP. We conclude that the dimerization defect is the molecular basis for perinatal HPP associated with the genotype G420S/G420S. PMID:23039266

Makita, Saori; Al-Shawafi, Hiba A; Sultana, Sara; Sohda, Miwa; Nomura, Shuichi; Oda, Kimimitsu

2012-12-01

86

Detection of endogenous alkaline phosphatase activity in intact cells by flow cytometry using the fluorogenic ELF-97 phosphatase substrate  

Science.gov (United States)

BACKGROUND: The alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (ELF((R))-97 for enzyme-labeled fluorescence) has been found useful for the histochemical detection of endogenous AP activity and AP-tagged proteins and oligonucleotide probes. In this study, we evaluated its effectiveness at detecting endogenous AP activity by flow cytometry. METHODS: The ELF-97 phosphatase substrate was used to detect endogenous AP activity in UMR-106 rat osteosarcoma cells and primary cultures of chick chondrocytes. Cells were labeled with the ELF-97 reagent and analyzed by flow cytometry using an argon ultraviolet (UV) laser. For comparison purposes, cells were also assayed for AP using a Fast Red Violet LB azo dye assay previously described for use in detecting AP activity by flow cytometry. RESULTS: The ELF-97 phosphatase substrate effectively detected endogenous AP activity in UMR-106 cells, with over 95% of the resulting fluorescent signal resulting from AP-specific activity (as determined by levamisole inhibition of AP activity). In contrast, less than 70% of the fluorescent signal from the Fast Red Violet LB (FRV) assay was AP-dependent, reflecting the high intrinsic fluorescence of the unreacted components. The ELF-97 phosphatase assay was also able to detect very low AP activity in chick chondrocytes that was undetectable by the azo dye method. CONCLUSIONS: The ELF-97 phosphatase assay was able to detect endogenous AP activity in fixed mammalian and avian cells by flow cytometry with superior sensitivity to previously described assays. This work also shows the applicability of ELF-97 to flow cytometry, supplementing its previously demonstrated histochemical applications. Copyright 1999 Wiley-Liss, Inc.

Telford, W. G.; Cox, W. G.; Stiner, D.; Singer, V. L.; Doty, S. B.

1999-01-01

87

Kidney bone disease and mortality in CKD: revisiting the role of vitamin D, calcimimetics, alkaline phosphatase, and minerals.  

Science.gov (United States)

Recent evidence suggests that the traditional syndromes known as renal osteodystrophy, secondary hyperparathyroidism, and vitamin D deficiency are related to mortality in persons with moderate to advanced chronic kidney disease (CKD). The so-called 'kidney bone disease', also known as 'mineral and bone disorders', is defined to include bone disorders, mineral disarrays, and vascular calcification. We have identified 14 common and clinically relevant conditions of contemporary nature that are related to the kidney bone disease, including calcitriol (active vitamin D) deficiency, 25(OH)-vitamin D deficiency, biochemical hyperparathyroidism, relatively low parathyroid hormone (PTH) level, increased serum alkaline phosphatase (hyperphosphatasemia), elevated fibroblast growth factor (FGF)-23, high turnover bone disease, adynamic bone disease, uremic osteoporosis, vascular calcification, hyper- and hypophosphatemia, and hyper- and hypocalcemia. We present a critical review of these 14 conditions with emphasis on patient survival and other pertinent clinical outcomes. We also review unresolved controversies surrounding the management of these conditions by administration of nutritional vitamin D (ergocalciferol and cholecalciferol), vitamin D receptor activators (calcitriol, alphacalcidiol, doxercalciferol), D-mimetics (paricalcitol, maxacalcitol), calcimimetics (cinacalcet), recombinant PTH (teriparatide), and receptor activator of nuclear factor-kappaB ligand modulators (denosumab); compare mortality predictability of PTH and alkaline phosphatase; and examine potential risks of bone disorders and mineral disarrays in CKD patients. PMID:20671739

Kalantar-Zadeh, Kamyar; Shah, Anuja; Duong, Uyen; Hechter, Rulin C; Dukkipati, Ramanath; Kovesdy, Csaba P

2010-08-01

88

Functional characterisation and transcript analysis of an alkaline phosphatase from the arbuscular mycorrhizal fungus Funneliformis mosseae.  

Science.gov (United States)

Alkaline phosphatases (ALP) in arbuscular mycorrhizal (AM) fungi have been suggested to be involved in transfer of phosphate from the mycorrhizal fungus to the host plant, but exact mechanisms are still unknown, partially due to the lack of molecular information. We isolated a full-length cDNA (FmALP) from the AM fungus Funneliformis mosseae (syn. Glomus mosseae) showing similarity with putative ALP genes from Rhizophagus intraradices (syn. Glomus intraradices) and Gigaspora margarita. For functional characterisation FmALP was expressed heterologously in the yeast Pichia pastoris. The recombinant FmALP protein had a pH optimum of 9.5, and catalysed the hydrolysis of glycerolphosphate and, to a lesser extent of glucose-1- and 6-phosphate, confirming it to be an alkaline phosphatase belonging to the family of alkaline phosphomonoesterases (EC 3.1.3.1). FmALP did not catalyse the hydrolysis of ATP or polyP. Relative FmALP transcript levels were analysed in intra- and extraradical hyphae isolated from F. mosseae infected ryegrass (Lolium perenne) using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). FmALP was highly expressed in intraradical hyphae at low P(i) supply, and its expression was repressed by high P(i) supply. Taken together this study provides evidence for mycorrhizal alkaline phosphatases playing a role in P mobilisation from organic substrates under P starvation conditions. PMID:23474124

Liu, Qianhe; Parsons, Anthony J; Xue, Hong; Jones, Chris S; Rasmussen, Susanne

2013-05-01

89

Cloning and Overexpression of Alkaline Phosphatase PhoK from Sphingomonas sp. Strain BSAR-1 for Bioprecipitation of Uranium from Alkaline Solutions?  

OpenAIRE

Cells of Sphingomonas sp. strain BSAR-1 constitutively expressed an alkaline phosphatase, which was also secreted in the extracellular medium. A null mutant lacking this alkaline phosphatase activity was isolated by Tn5 random mutagenesis. The corresponding gene, designated phoK, was cloned and overexpressed in Escherichia coli strain BL21(DE3). The resultant E. coli strain EK4 overexpressed cellular activity 55 times higher and secreted extracellular PhoK activity 13 times higher than did BS...

Nilgiriwala, Kayzad S.; Alahari, Anuradha; Rao, Amara Sambasiva; Apte, Shree Kumar

2008-01-01

90

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

Directory of Open Access Journals (Sweden)

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

Deepak Ganjewala

2010-01-01

91

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

Energy Technology Data Exchange (ETDEWEB)

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

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

1998-01-01

92

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

CERN Document Server

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

Berezhetskyy, A

2008-01-01

93

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

Directory of Open Access Journals (Sweden)

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

Igwe CU

2004-10-01

94

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

Science.gov (United States)

Determination of phosphate ions concentration is very important from both, environmental and clinical point of view. In this study, a simple and novel conductometric biosensor for indirect determination of the phosphate ions in aqueous solution has been developed. The developed biosensor is based on the inhibition of immobilized alkaline phosphatase activity, in the presence of the phosphate ions. This is the first time we developed a mono-enzymatic biosensor for indirect estimation of phosphate ions. The developed biosensor showed a broad linear response (as compared to other reported biosensors) for phosphate ions in the range of 0.5-5.0mM (correlation coefficient=0.995), with a detection limit of 50µM. Different optimized parameters were obtained as the buffer concentration of 30mM, substrate concentration of 1.0mM, and a pH of 9.0. All the optimized parameters were analyzed by analysis of variance, and were found to be statistically significant at a level of ?=0.05. The developed biosensor is also suitable to determine the serum phosphate concentration, with a recovery of 86-104%, while a recovery of 102% was obtained from the water samples that were spiked with 500µM phosphate. A relative standard deviation in the conductance response for five successive measurements (n=5) did not exceed 7%, with a shelf life of 30 days. With a lower detection limit and a higher recovery, the biosensor provides a facile approach for phosphate estimation in biological fluids. PMID:25656777

Upadhyay, Lata Sheo Bachan; Verma, Nishant

2015-06-15

95

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

OpenAIRE

Placental alkaline phosphatase (PLAP) is a promising ovarian cancer biomarker. Here, we describe the isolation, affinity-maturation and characterization of two fully human monoclonal antibodies (termed B10 and D9) able to bind to human PLAP with a dissociation constant (Kd) of 10 and 30 nM, respectively. The ability of B10 and D9 antibodies to recognize the native antigen was confirmed by Biacore analysis, FACS and immunofluorescence studies using ovarian cancer cell lines and freshly-frozen ...

Ravenni, Niccolo?; Weber, Marcel; Neri, Dario

2013-01-01

96

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

OpenAIRE

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) ...

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

2013-01-01

97

In situ hybridization of cytokine mRNA using alkaline phosphatase-labelled oligodeoxynucleotide probes  

DEFF Research Database (Denmark)

In situ hybridization is a powerful tool for visualizing cellular gene expression in morphologically preserved brain tissue giving precise information on the regional expression of specific mRNA sequences in cells of diverse phenotype. Here, we describe a sensitive, simple, and robust method using alkaline phosphatase (AP)-labelled oligodeoxynucleotide probes to detect cytokine mRNA in the acutely injured or inflamed mouse CNS.

Clausen, Bettina Hjelm; Fenger, Christina

2013-01-01

98

Expression of the gene encoding secreted placental alkaline phosphatase (SEAP) by a nondefective adenovirus vector.  

Science.gov (United States)

A nondefective recombinant human adenovirus 5 (Ad5) carrying the SEAP gene, encoding human secreted placental alkaline phosphatase, in the E3 region of the Ad5 genome was obtained. The expression of SEAP at the early and late stages of Ad5 infection was demonstrated in permissive and semi-permissive cell cultures. The amount of SEAP in the culture medium of the 293 cells was 13.6% of the total protein. PMID:8482541

Doronin, K K; Zakharchuk, A N; Grinenko, N F; Yurov, G K; Krougliak, V A; Naroditsky, B S

1993-04-30

99

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

OpenAIRE

In order to clarify the structural basis of the halophilic characteristics of an alkaline phosphatase derived from the moderate halophile Halomonas sp. 593 (HaAP), the tertiary structure of HaAP was determined to 2.1?Å resolution by X-ray crystallography. The structural properties of surface negative charge and core hydrophobicity were shown to be intermediate between those characteristic of halophiles and non-halophiles, and may explain the unique functional adaptation to a wide range of ...

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

2014-01-01

100

Alkaline Phosphatase and Other Hydrolyases Produced by Cenococcum graniforme, an Ectomycorrhizal Fungus  

OpenAIRE

Cell extracts of Cenococcum graniforme have been found to contain the following hydrolytic enzymes: protease, esterase, ?-d-galactopyranosidase, ?-d-galactopyranosidase, ?-d-mannopyranosidase, ?-d-xylopyranosidase, ?-d-glucopyranosidase, ?-d-glucopyranosidase, and alkaline phosphatase. Sulfatase, inorganic pyrophosphatase, and ?-d-mannopyranosidase were not detected in the extracts. ?-d-Xylopyranosidase and ?-d-mannopyranosidase were most active in the neutral pH range, protease and ...

Bae, Kwang-sung; Barton, Larry L.

1989-01-01

101

Measurement of placental alkaline phosphatase activity in benign and malignant pleural effusions.  

OpenAIRE

The usefulness of placental alkaline phosphatase (PLAP) as a diagnostic marker of malignancy was assessed in pleural fluid from 60 patients with effusions. Pleural fluid PLAP activities were measured by an enzyme linked immunoassay (ELISA) using the two monoclonal antibodies H17E2 and H317. Similar values were found in groups of patients with primary bronchial tumours (n = 12), secondary malignancies (n = 23), and "benign" conditions (n = 25). The highest values were found in a small subgroup...

Fergusson, R. J.; Fisken, J.; Mcintyre, M. A.; Roulston, J. E.; Leonard, R. C.

1992-01-01

102

[Effectiveness of methods of isolating specific class-G rabbit antibodies for the immunoenzyme determination of human placental alkaline phosphatase].  

Science.gov (United States)

Four different chromatographic methods of IgG isolation from rabbit antisera to placental alkaline phosphatase (HPAP) have been compared. The antibodies were obtained by ion-exchange chromatography and affinity chromatography on protein-A-sepharose, on the sepharose with immobilized antigen. IgG samples were characterized by the content of specific antibodies to HPAP and checked in enzyme immunoassay (EIA). IgG purified on immobilized antigen were found to be the optimal both from the point of view of the specific antibodies content and EIA sensitivity, but satisfactory results could be also obtained with ion-exchange and protein-A-chromatography purified IgG. The last two isolation methods are simpler and provide 3-10 ng/ml sensitivity of HPAP detection, which is lower, as compared with the test employing affinity antibodies (1 ng/ml), but allows the detection of HPAP in serum samples. PMID:3046673

Voronov, A V; Gudima, S O; Mierinia, A A; Vengerov, Iu Iu; Votrin, I I

1988-08-01

103

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

1986-01-01

104

Chemiluminescence-based pesticide biosensor utilizing the intelligent evolved properties of the enzyme alkaline phosphatase  

Energy Technology Data Exchange (ETDEWEB)

A methodology is described for immobilizing the enzyme alkaline phosphatase onto a glass surface using a novel biotinylated copolymer, poly(3-undecylthiophene-co-3- methanoithiophene). A streptavidin conjugate of alkaline phosphatase is used in this study. The biotinylated polymer is attached to the silanized glass surface via hydrophobic interactions and the enzyme is interfaced with the polymer through the classical biotin- streptavidin interaction. Alkaline phosphatase catalyzes the dephosphorylation of a macrocyclic compound, chloro-3-(4-methoxy spiro) (1,2 dioxetane-3-2`-tricyclo-) (3.3.1.1 )-(decani-4-yl) phenyl phosphate, to a species which emits energy by chemiluminescence. This chemiluminescence signal can be detected with a photomultiplier tube for enzymatic catalysis with the biocatalyst both in solution and immobilized on a glass surface. The signal generation is inhibited by the organophosphorus based insecticides such as paraoxon as well as nerve agents. We demonstrate in this study that a number of organophosphorus based insecticides inhibit the enzyme-mediated generation of chemiluminescence signal. This is true for the enzyme conjugate both free in solution and immobilized on a glass surface. In solution, the inhibition resembles the case of a partially uncompetitive system. By this type of inhibition we are able to detect pesticides down to about 50 ppb for the enzyme in solution. The pesticide detection limit of immobilized enzyme is currently being investigated. The enzyme is capable of a number of measurement cycles without significant loss of signal level.

Ayyagari, M.; Kamtekar, S.; Pande, R.; Marx, K.; Kumar, J.

1994-12-31

105

Alkaline phosphatases in microbialites and bacterioplankton from Alchichica soda lake, Mexico.  

Science.gov (United States)

Dissolved organic phosphorus utilization by different members of natural communities has been closely linked to microbial alkaline phosphatases whose affiliation and diversity is largely unknown. Here we assessed genetic diversity of bacterial alkaline phosphatases phoX and phoD, using highly diverse microbial consortia (microbialites and bacterioplankton) as study models. These microbial consortia are found in an oligo-mesotrophic soda lake with a particular geochemistry, exhibiting a low calcium concentration and a high Mg : Ca ratio relative to seawater. In spite of the relative low calcium concentration in the studied system, our results highlight the diversity of calcium-based metallophosphatases phoX and phoD-like in heterotrophic bacteria of microbialites and bacterioplankton, where phoX was the most abundant alkaline phosphatase found. phoX and phoD-like phylotypes were more numerous in microbialites than in bacterioplankton. A larger potential community for DOP utilization in microbialites was consistent with the TN : TP ratio, suggesting P limitation within these assemblages. A cross-system comparison indicated that diversity of phoX in Lake Alchichica was similar to that of other aquatic systems with a naturally contrasting ionic composition and trophic state, although no phylotypes were shared among systems. PMID:25112496

Valdespino-Castillo, Patricia M; Alcántara-Hernández, Rocio J; Alcocer, Javier; Merino-Ibarra, Martín; Macek, Miroslav; Falcón, Luisa I

2014-11-01

106

Cobalt(III), a probe of metal binding sites of Escherichia coli alkaline phosphatase.  

Science.gov (United States)

To facilitate the study of individual metal binding sites of polymeric metalloproteins, conversion of exchange-labile Co(II) in E. coli alkaline phosphatase (EC 3.1.3.1) to exchange-inert Co(III) was examined. Oxidation of Co(II) alkaline phosphatase with hydrogen peroxide results in a single absorption maximum at 530 nm and loss both of the characteristic electron paramagnetic signal and of enzymatic activity. Zinc neither reactivates this enzyme nor displaces the oxidized cobalt atoms. Metal and amino-acid analyses demonstrate that oxidation alters neither cobalt binding nor amino-acid composition of the enzyme. Al data are consistent with the conclusion that hydrogen peroxide oxidizes Co(II) in alkaline phosphatase to Co(III). Polymeric metalloenzymes can contain different categories of metal atoms serving in catalysis, structure stabilization, and/or control and exerting their effects independently or interdependently. The in situ conversion of exchange-labile Co(II) to exchange-stable (Co(III) offers a method to selectively and differentially "freeze" cobalt atoms at their respective binding sites. The accompanying spectral changes and concomitant retardation in ligand exchange reactions may be used to differentiate between specific metal binding sites that serve different roles in polymeric metalloenzymes. PMID:164026

Anderson, R A; Vallee, B L

1975-01-01

107

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

International Nuclear Information System (INIS)

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

108

Differential alkaline phosphatase responses of rat and human bone marrow derived mesenchymal stem cells to 45S5 bioactive glass  

OpenAIRE

Bioactive glass is used as both a bone filler and as a coating on implants, and has been advocated as a potential osteogenic scaffold for tissue engineering. Rat derived mesenchymal stem cells (MSCs) show elevated levels of levels of alkaline phosphatase activity when grown on 45S5 bioactive glass as compared to standard tissue culture plastic. Similarly, exposure to the dissolution products of 45S5 elevates alkaline phosphatase activity and other osteogenic markers in these cells. We investi...

Reilly, Gwendolen C.; Radin, Shula; Chen, Andrew T.; Ducheyne, Paul

2007-01-01

109

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

2009-04-01

110

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

International Nuclear Information System (INIS)

We describe radioimmunoassay for human prostatic acid phosphatase [orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2] in serum, with use of monospecific antisera raised in rabbits against highly purified acid phosphatase from human prostates. The antiserum did not cross react with partly purified acid phosphatases from human spleen, erythrocytes, or synovial tissues. 125I-labeled acid phosphatase was prepared by a Chloramine T method, and the bound and free antigen was separated in the assay by use of anti-rabbit gamma-globulin raised in sheep. Uniform low nonspecific binding of the [125I]acid phosphatase was achieved by using acid-phosphatase-free serum to prepare standard curves and diluted samples of serum with high acid phosphatase activities. Concentrations of immunoreactive acid phosphatase in the serum of healthy men ranged from <1 to 10 ?g/liter and for 12 patients with advanced prostatic carcinoma between 100 and 500 ?g/liter. The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group

111

Diagnosis of prostate cancer using a radioimmunoassay for prostatic acid phosphatase in serum  

International Nuclear Information System (INIS)

The paper describes the development and evaluation of a specific radioimmunoassay for the determination of prostatic acid phosphatase in serum as a useful aid in the detection of prostatic cancer. (Auth.)

112

[An immunochemical study of M-HeLa cells as producers of human placental alkaline phosphatase].  

Science.gov (United States)

While analyzing M-HeLa cells by IFA technique secretory and membrane-bound forms of human placental alkaline phosphatase (HPAP) were detected. Activity of secretory HPAP increased if cell density and incubation time were increased too. After short influence of heat shock (15 min at 42 degrees C) activity of secretory HPAP increased for 45% and intracellular HPAP 3 for 37%. It is proposed that HPAP take part in organization of first response to heat shock and support cellular thermotolerance. PMID:2194582

Gudima, S O; Liakhov, V V; Sokolov, A V; Terekhov, O P; Serov, S M; Sukhikh, G T; Vengerov, Iu Iu

1990-03-01

113

Cloning, sequencing, and chromosomal localization of human term placental alkaline phosphatase cDNA.  

OpenAIRE

A human term (third trimester) placental alkaline phosphatase (PLAP; EC 3.1.3.1) cDNA was isolated from a human placental lambda gt11 cDNA library. The expression library was screened by using rabbit antibodies against PLAP and oligonucleotide probes. DNA sequence analysis of a positive clone with an insert of 2.7 kilobase pairs allowed us to predict the complete amino acid sequence of PLAP (530 residues), which coincided with the reported 42 N-terminal amino acid sequence of PLAP except at p...

Kam, W.; Clauser, E.; Kim, Y. S.; Kan, Y. W.; Rutter, W. J.

1985-01-01

114

Zinc status and vacuolar zinc transporters control alkaline phosphatase accumulation and activity in Saccharomyces cerevisiae  

OpenAIRE

Little is known about how metalloproteins in the secretory pathway obtain their metal ion cofactors. We used the Pho8 alkaline phosphatase of the yeast Saccharomyces cerevisiae to probe this process in vivo. We found that both Pho8 activity and protein accumulation are zinc dependent and decrease in zinc-limited cells. Low Pho8 accumulation was the result of degradation by vacuolar proteases. Surprisingly, the protective effect of zinc on Pho8 stability was not solely due to Zn2+ binding to t...

Qiao, Wei; Ellis, Charissa; Steffen, Janet; Wu, Chang-yi; Eide, David J.

2009-01-01

115

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

OpenAIRE

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

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

2000-01-01

116

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

OpenAIRE

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 Akp...

Lei, Wei; Nguyen, Heidi; Brown, Naoko; Ni, Hua; Kiffer-moreira, Tina; Reese, Jeff; Milla?n, Jose? Luis; Paria, Bibhash C.

2013-01-01

117

Comparative effectiveness of the cholera toxin B subunit and alkaline phosphatase as carriers for oral vaccines.  

OpenAIRE

The purpose of this study was to determine whether the B subunit of cholera toxin (CtxB) has adjuvant activity over and above serving as a carrier protein for orally administered vaccines. An oligonucleotide that encodes an antigenic determinant (GtfB.1) from the glucosyltransferase B gene (gtfB) of Streptococcus mutans was genetically fused to the 5' terminus of either the CtxB gene (ctxB) or the Escherichia coli alkaline phosphatase gene (phoA). The resulting chimeric proteins were expresse...

Dertzbaugh, M. T.; Elson, C. O.

1993-01-01

118

Alkaline phosphatase from the hyperthermophilic bacterium T. maritima requires cobalt for activity.  

Science.gov (United States)

The hyperthermophilic bacterium Thermotoga maritima encodes a gene sharing sequence similarities with several known genes for alkaline phosphatase (AP). The putative gene was isolated and the corresponding protein expressed in Escherichia coli, with and without a predicted signal sequence. The recombinant protein showed phosphatase activity toward the substrate p-nitrophenyl-phosphate with a k(cat) of 16 s(-1) and a K(m) of 175 microM at a pH optimum of 8.0 when assayed at 25 degrees C. T. maritima phosphatase activity increased at high temperatures, reaching a maximum k(cat) of 100 s(-1), with a K(m) of 93 microM at 65 degrees C. Activity was stable at 65 degrees C for >24 h and at 90 degrees C for 5 h. Phosphatase activity was dependent on divalent metal ions, specifically Co(II) and Mg(II). Circular dichroism spectra showed that the enzyme gains secondary structure on addition of these metals. Zinc, the most common divalent metal ion required for activity in known APs, was shown to inhibit the T. maritima phosphatase enzyme at concentrations above 0.3 moles Zn: 1 mole monomer. All activity was abolished in the presence of 0.1 mM EDTA. The T. maritima AP primary sequence is 28% identical when compared with E. coli AP. Based on a structural model, the active sites are superimposable except for two residues near the E. coli AP Mg binding site, D153 and K328 (E. coli numbering) corresponding to histidine and tryptophan in T. maritima AP, respectively. Sucrose-density gradient sedimentation experiments showed that the protein exists in several quaternary forms predominated by an octamer. PMID:11910033

Wojciechowski, Cheryl L; Cardia, James P; Kantrowitz, Evan R

2002-04-01

119

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

120

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

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

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

2013-12-15

121

Transformation of glucocorticoid receptors bound to the antagonist RU 486: Effects of alkaline phosphatase  

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RU 486 is a synthetic steroid that binds avidly to glucocorticoid receptors without promoting their transformation into activated transcription factors. A significant part of this behavior has been shown to be due to a failure of the RU 486 bound receptor to be efficiently released from a larger (sedimenting at 8-9 S) multimeric complex containing the 90-kDa heat shock protein. The studies have found that in vitro at 15{degree}C the RU 486-receptor was slowly released from the 8-9S complex and converted into a DNA binding protein by a process that could be blocked by sodium fluoride. Moreover, this transition was significantly accelerated by treatment with alkaline phosphatase. High-resolution anion-exchange chromatography showed that the profile of receptor subspecies released from the 8-9S complex was different for the RU 486 bound receptor when compared to the receptor occupied by the agonist triamcinolone acetonide. Production of the earliest eluting receptor form (peak A) was inhibited with RU 486. Treatment of the Ru 486-receptor with alkaline phosphatase increased the formation of the peak A subspecies as well as the capacity of receptor to bind DNA-cellulose. Taken together, the results indicate that phosphorylation of the receptor or a tightly bound factor contributes to defining the capacity with which individual steroids can promote dissociation of the 8-9S complex and conversion of the glucocorticoid receptor into a DNA-binding protein.

Gruol, D.J.; Wolfe, K.A. (Salk Institute for Biological Studies, San Diego, CA (USA))

1990-08-28

122

Alkaline phosphatase in osteoblasts is down-regulated by pulsatile fluid flow  

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It is our hypothesis that interstitial fluid flow plays a role in the bone remodeling response to mechanical loading. The fluid flow-induced expression of three proteins (collagen, osteopontin, and alkaline phosphatase) involved in bone remodeling was investigated. Rat calvarial osteoblasts subjected to pulsatile fluid flow at an average shear stress of 5 dyne/cm2 showed decreased alkaline phosphatase (AP) mRNA expression after only 1 hour of flow. After 3 hours of flow, AP mRNA levels had decreased to 30% of stationary control levels and remained at this level for an additional 5 hours of flow. Steady flow (4 dyne/cm2 fluid shear stress), in contrast, resulted in a delayed and less dramatic decrease in AP mRNA expression to 63% of control levels after 8 hours of flow. The reduced AP mRNA expression under pulsatile flow conditions was followed by reduced AP enzyme activity after 24 hours. No changes in collagen or osteopontin mRNA expression were detected over 8 hours of pulsatile flow. This is the first time fluid flow has been shown to affect gene expression in osteoblasts.

Hillsley, M. V.; Frangos, J. A.

1997-01-01

123

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

2010-10-01

124

Expression of a human placental alkaline phosphatase gene in transfected cells: Use as a reporter for studies of gene expression  

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The human placental alkaline phosphatase gene has been cloned and reintroduced into mammalian cells. When a plasmid carrying the gene under control of the simian virus 40 early promoter (pSV2Apap) is transfected into a variety of different cell types, placental alkaline phosphatase activity can readily be detected by using whole cell suspensions or cell lysates. Alkaline phosphatase activity can also be visualized directly in individual transfected cells by histochemical staining. The gene is appropriate for use as a reporter in studies of gene regulation since its expression is dependent on the presence of exogenous transcription control elements. The overall assay to detect the expression of the gene is quantitative, very rapid, and inexpensive. Cotransfections of cells with pSV2Apap and a related plasmid carrying the bacterial chloramphenicol acetyltransferase gene (pSV2Acat) indicate that transcription of these two genes is detected with roughly the same sensitivity.

Henthorn, P.; Zervos, P.; Raducha, M.; Harris, H.; Kadesch, T.

1988-09-01

125

Alkaline and Acid Phosphatase Activity in Blood Plasma of Chickens Irradiated by Low dose Gamma Radiation  

International Nuclear Information System (INIS)

In our previous paper (Kraljevic et, al, 2000; Kraljevic et al 2002) we showed that the growth of the chickens hatched from eggs irradiated with 0.15 Gy gamma-rays before incubation was significantly higher than in controls during the fattening period (1-42 days). The concentration of total protein, glucose and cholesterol in the blood plasma of the same chickens was also significantly changed. In this paper an attempt was made to determine the effect of irradiation of eggs by low dose ionizing radiation before incubation upon activity of alkaline and acid phosphatase in the blood plasma of chickens hatched from irradiated eggs. The eggs of heavy breeding chickens were irradiated by dose of 0.15 Gy gamma radiation (60 Co) before incubation. Along with the chickens which were hatched from irradiated eggs, there was a control group of chickens hatched from nonirradiated eggs. All other conditions were the same for both groups. After hatching, blood samples were taken from the wing vein on days 1, 3, 5, 6, 10, 20, 30 and 42. The activity of both enzymes was determined spectrophotometrically by using Boehring Mannheim GmbH optimized kits. the activity of alkaline phosphatase in blood plasma was decreased on days 42, and the activity of acid phosphatase in the blood plasma of the same chickens was increased on day 42. Obtained results confirm our early obtained results that low dose of gamma radiation has effects upon metabolic processes in the chickens hatched from eggs icesses in the chickens hatched from eggs irradiated before incubation. (Author)

126

Endothelial alkaline phosphatase activity loss as an early stage in the development of radiation-induced heart disease in rats  

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Alkaline phosphatase activity of capillary endothelial cells in the heart of Wistar and Sprague-Dawley rats was studied sequentially after single doses of 10, 15, 20, or 25 Gy. Following irradiation capillary density and alkaline phosphatase activity were focally lost before myocardial degeneration or clinical symptoms of heart disease developed. Recovery from both changes took place after doses of 10 or 15 Gy. The decrease in capillary density and enzyme activity showed the same strain difference in latency times and in the extent of the lesions as previously described for pathological and clinical signs of heart disease

127

A Novel Bifunctional Hybrid with Marine Bacterium Alkaline Phosphatase and Far Eastern Holothurian Mannan-Binding Lectin Activities  

OpenAIRE

A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa...

Balabanova, Larissa; Golotin, Vasily; Kovalchuk, Svetlana; Bulgakov, Alexander; Likhatskaya, Galina; Son, Oksana; Rasskazov, Valery

2014-01-01

128

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

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

129

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 Mg2+.

130

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

131

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

132

?? Adrenergic Receptor Activation Suppresses Bone Morphogenetic Protein (BMP)-Induced Alkaline Phosphatase Expression in Osteoblast-Like MC3T3E1 Cells.  

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? adrenergic stimulation suppresses bone formation in vivo while its actions in osteoblastic differentiation are still incompletely understood. We therefore examined the effects of ?2 adrenergic stimulation on osteoblast-like MC3T3-E1 cells focusing on BMP-induced alkaline phosphatase expression. Morphologically, isoproterenol treatment suppresses BMP-induced increase in the numbers of alkaline phosphatase-positive small foci in the cultures of MC3T3-E1 cells. Biochemically, isoproterenol treatment suppresses BMP-induced enzymatic activity of alkaline phosphatase in a dose-dependent manner. Isoproterenol suppression of alkaline phosphatase activity is observed even when the cells are treated with high concentrations of BMP. With respect to cell density, isoproterenol treatment tends to suppress BMP-induced increase in alkaline phosphatase expression more in osteoblasts cultured at higher cell density. In terms of treatment protocol, continuous isoproterenol treatment is compared to cyclic treatment. Continuous isoproterenol treatment is more suppressive against BMP-induced increase in alkaline phosphatase expression than cyclic regimen. At molecular level, isoproterenol treatment suppresses BMP-induced enhancement of alkaline phosphatase mRNA expression. Regarding the mode of isoproterenol action, isoproterenol suppresses BMP-induced BRE-luciferase activity. These data indicate that isoproterenol regulates BMP-induced alkaline phosphatase expression in osteoblast-like MC3T3E1 cells. J. Cell. Biochem. 116: 1144-1152, 2015. © 2014 Wiley Periodicals, Inc. PMID:25536656

Yamada, Takayuki; Ezura, Yoichi; Hayata, Tadayoshi; Moriya, Shuichi; Shirakawa, Jumpei; Notomi, Takuya; Arayal, Smriti; Kawasaki, Makiri; Izu, Yayoi; Harada, Kiyoshi; Noda, Masaki

2015-06-01

133

Expression, secretion and surface display of a human alkaline phosphatase by the ciliate Tetrahymena thermophila  

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Full Text Available Abstract Background Tetrahymena thermophila possesses many attributes that render it an attractive host for the expression of recombinant proteins. Surface proteins from the parasites Ichthyophthirius multifiliis and Plasmodium falciparum and avian influenza virus antigen H5N1 were displayed on the cell membrane of this ciliate. Furthermore, it has been demonstrated that T. thermophila is also able to produce a functional human DNase I. The present study investigates the heterologous expression of the functional human intestinal alkaline phosphatase (hiAP using T. thermophila and thereby presents a powerful tool for the optimization of the ciliate-based expression system. Results Functional and full length human intestinal alkaline phosphatase was expressed by T. thermophila using a codon-adapted gene containing the native signal-peptide and GPI (Glycosylphosphatidylinositol anchor attachment signal. HiAP activity in the cell extract of transformants suggested that the hiAP gene was successfully expressed. Furthermore, it was demonstrated that the enzyme was modified with N-glycosylation and localized on the surface membrane by the C-terminal GPI anchor. A C-terminally truncated version of hiAP lacking the GPI anchor signal peptide was secreted into the medium as an active enzyme. In a first approach to establish a high level expression system up to 14,000 U/liter were produced in a time frame of two days, which exceeds the production rate of other published expression systems for this enzyme. Conclusions With the expression of hiAP, not only a protein of commercial interest could be produced, but also a reporter enzyme that offers the possibility to analyze T. thermophila genes that play a role in the regulation of protein secretion. Additionally, the fact that ciliates do not secrete an endogenous alkaline phosphatase provides the possibility to use the truncated hiAP as a reporter enzyme, allowing the quantification of measures that will be necessary for further optimization of the host strains and the fermentation processes.

Herrmann Lutz

2011-01-01

134

Stabilization of alkaline phosphatase with Au@Ag2O nanoparticles.  

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Here, we report that a conductive Au@Ag(2)O nanoparticle structure significantly enhances the stability of alkaline phosphatase (AlkP) in the presence of the inhibitors urea and l-phenylalanine (Phe). The enzyme/nanoparticle construct is prepared by associating the enzyme with citrate-capped Au particles, and then adding Ag(+). UV-vis and XPS spectroscopy and transmission electron microscopy confirm the core@shell structure. AlkP activity was quantified in the presence and absence of the two inhibitors using a time-resolved colorimetric assay. The results indicate that 21% of the initial active AlkP is incorporated into the nanoparticle structure. More importantly, however, the Au@Ag(2)O core@shell host reduces the inhibitory effect of urea and Phe by factors ranging from 3 to 12, depending on the inhibitor and its concentration, compared to the wild-type enzyme. PMID:21846098

Zaccheo, Brian A; Crooks, Richard M

2011-09-20

135

[Thermostable placental-type alkaline phosphatase in the malignantly degenerated endometrium].  

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A comparative immunochemical study established that heat-stable placental alkaline phosphatase (HPAP) is contained only in malignant epithelium and is not present in the tissues of fetal and definitive uterus. The heterogeneity of HPAP was shown by means of electrophoresis. There is a direct correlation between HPAP level and the degree of tumor differentiation. Immunohistochemical and histochemical studies showed that HPAP occurs in areas of tumor where glandular structure is most pronounced. HPAP secretion in endometrial mucus was observed. An immunochemical test for HPAP is suggested which may be used in evaluating the degree of tumor differentiation and selecting a suitable scheme of treatment. Immunochemical assay of endometrial mucus for HPAP may offer good advantage in mass screenings of females at risk. PMID:7090293

Volodin, M A; Afanas'eva, A V; Bokhman, Ia V; Chepik, O F; Shvarev, E G

1982-01-01

136

A real-time fluorescent assay for the detection of alkaline phosphatase activity based on carbon quantum dots.  

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A convenient and real-time fluorometric assay with the assistance of copper ions based on aggregation and disaggregation of carbon quantum dots (CQDs) was developed to achieve highly sensitive detection of alkaline phosphatase activity. CQDs and pyrophosphate anions (PPi) were used as the fluorescent indicator and substrate for ALP activity assessment respectively. Richness of carboxyl groups on the surface of CQDs enables their severe aggregation triggered by copper ions, which results in effective fluorescence quenching. Under the catalytic hydrolysis of ALP, PPi can be rapidly transformed to phosphate ions. Stronger affinity of phosphate ions to copper ions than carboxyl groups is taken advantage of to achieve fluorescence recovery induced by re-dispersion of CQDs in the presence of ALP and PPi. Quantitative evaluation of ALP activity in a broad range from 16.7 to 782.6U/L with the detection limit of 1.1U/L can be realized in this way, which endows the assay with high enough sensitivity for practical detection in human serum. This strategy broadens the sensing application of fluorescent CQDs with excellent biocompatibility, and provides an example based on disaggregation in optical probe development. PMID:25660658

Qian, Zhao Sheng; Chai, Lu Jing; Huang, Yuan Yuan; Tang, Cong; Jia Shen, Jia; Chen, Jian Rong; Feng, Hui

2015-06-15

137

Carbon quantum dots-based recyclable real-time fluorescence assay for alkaline phosphatase with adenosine triphosphate as substrate.  

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A convenient, reliable, and highly sensitive real-time assay for alkaline phosphatase (ALP) activity in the continuous and recyclable way is established on the basis of aggregation and disaggregation of carbon quantum dots (CQDs) through the competitive assay approach. CQDs and adenosine triphosphate (ATP) were used as the fluorescent indicator and substrate for ALP activity assessment, respectively. Richness of carboxyl groups on the surface of CQDs enables their severe aggregation triggered by cerium ions, which results in effective fluorescence quenching. Under the catalytic hydrolysis of ALP, ATP can be rapidly transformed to phosphate ions. Stronger affinity of phosphate ions to cerium ions than carboxyl groups is taken advantage of to achieve fluorescence recovery induced by redispersion of CQDs in the presence of ALP and ATP. Quantitative evaluation of ALP activity in a broad range from 4.6 to 383.3 U/L with the detection limit of 1.4 U/L can be realized in this way, which endows the assay with high enough sensitivity for practical detection in human serum. The assay can be used in a recyclable way for more than three times since the generated product CePO4 as a precipitate can be easily removed from the standard assay system. This strategy broadens the sensing application of fluorescent CQDs with excellent biocompatibility and provides an example based on disaggregation in optical probe development. PMID:25642736

Qian, Zhaosheng; Chai, Lujing; Tang, Cong; Huang, Yuanyuan; Chen, Jianrong; Feng, Hui

2015-03-01

138

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

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

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

1987-07-01

139

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

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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

140

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)

141

Properties of Na+/K+ ATPase and alkaline phosphatase alter during spontaneous and radiation-induced leukemogenesis in mice  

International Nuclear Information System (INIS)

Properties are characterized of Na+/K+ ATPase and alkaline phosphatase in thymocytes or thymoblasts from mice of two strains: AKR in which thymoma developed spontaneously, and C57Bl in which the development was induced by X-irradiation (total dose: 5.4 Gy in 3 fractions). It was found that before thymoma could be discerned morphologically the properties of the two enzymes changed. There was a decrease in 86Rb uptake and in the rate of ATP hydrolysis per cell (both strains) as well as an increase in alkaline phosphatase activity per cell (C57Bl mice). In both spontaneous and radiation-induced thymomas 86Rb uptake, ATP hydrolysis and 3H-ouabain binding per cell were higher than in normal thymuses. Likewise, alkaline phosphatase activity per cell was higher in the thymomas than in the thymuses; this increase was accompanied by the appearance of additional isoenzyme(s) (1 in AKR, 2 in C57Bl). These changes were compared with cAMP content and 3H-thymidine incorporation, taken as indicators of the proliferative activity, and their high correlation in both AKR and C57Bl mice allowed to distinguish a pre-leukemic period. In that period thymoblasts clearly differed from the normal ones in Na+/K+ ATPase and alkaline phosphatase properties as well as proliferation, although the morphology of the thymus was still unchanged. (author)

142

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

143

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

Science.gov (United States)

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

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

2013-08-01

144

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

Science.gov (United States)

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

Chung, Thomas D Y

2013-01-01

145

Characterization of Alkaline Phosphatase Producing Bacteria Isolated from Thai Fermented Fish Products  

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Full Text Available Seventeen isolates of alkaline phosphatase (ALP producing bacteria were screened and systematically studied. They were divided into 10 groups on the basis of their phenotypic characteristics and 16S rRNA gene sequence analyses. The Gram-positive coccal isolates in Group I (3 isolates, Group II (3 isolates and Group III (1 isolate showed 99.2%, 99.5-99.7% and 99.8% sequences similarity to Staphylococcus saprophyticus, S. napalensis and S. sciuri, respectively. Each Gram-positive rod-shaped isolates in Group IV and V belonged to the genus Bacillus and was closely related to B. vietnamensis and B. safensis with 99.2% and 99.4% sequences similarity, respectively. The Gram-positive, moderately halophilic rod-shaped bacteria in Group VI (3 isolates, Group VII (1 isolate, Group VIII (1 isolate and Group IX (2 isolates were closely related to Virgibacillus halodenitrificans (98.5-99.1%, Oceanobacillus iheyensis (99.3%, Halobacillus mangrove (97.9% and H. dabanensis (98.5-98.6%, respectively. One Gram-negative isolate of moderately halophilic bacterium (Group X was closely related to Idiomarina zobelli (98.0%. They possessed phosphatase activities ranged from 10.08-70.96 U ml-1. The isolate NSW 13-2 (Group VI showed the highest ALP. 

Jaruwan Sitdhipol

2012-05-01

146

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

Science.gov (United States)

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

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

2011-12-01

147

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

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

Praveen Kumar Vemuri et al

2012-09-01

148

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

International Nuclear Information System (INIS)

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

149

Schistosoma mansoni: molecular characterization of Alkaline Phosphatase and expression patterns across life cycle stages.  

Science.gov (United States)

Here we describe the cloning and characterization of the Schistosoma mansoni Alkaline Phosphatase (SmAP), previously identified in the tegument of adult worms. SmAP encodes a complete sequence composed of 536 amino acids containing an N-terminal signal peptide, five N-glycosylation sites, and a GPI anchor signal, similar to that described for mammalian orthologs. Real-time RT-PCR and Western blot experiments suggest a rapid translation as soon as cercariae are transformed into schistosomula. Immunolocalization analysis shows that the protein is widely distributed in the worm tissues, with increased concentration in the vitelline glands of female parasites. Furthermore, the surface localization of this enzyme was quantitatively supported by its enzymatic activity in live ex vivo or cultured parasites throughout the life cycle stages. The fact that cercariae accumulate large amounts of SmAP mRNA, which rapidly translates into protein upon schistosomula transformation, indicates it may have an important role in host invasion. PMID:21784070

Araujo-Montoya, B O; Rofatto, H K; Tararam, C A; Farias, L P; Oliveira, K C; Verjovski-Almeida, S; Wilson, R A; Leite, L C C

2011-11-01

150

Identification of enterotoxigenic Escherichia coli with synthetic alkaline phosphatase-conjugated oligonucleotide DNA probes  

International Nuclear Information System (INIS)

Alkaline phosphatase-conjugated (AP) 26-base oligonucleotide DNA probes were compared with the same probes labeled with gamma-32P for the identification of heat-labile (LT) and heat-stable (ST) enterotoxigenic Escherichia coli (ETEC). The AP oligonucleotide probes were as sensitive as the radiolabeled (RL) probes in detecting LT and STA-2 target cell DNA, but the AP ST probe, which differed from STA-1 by two bases, was less sensitive than the RL probe in detecting STA-1 DNA (6.25 versus 0.78 ng). Of 94 ETEC that were identified with the RL probe, the AP probes detected 93% (28 of 30) of ST, 73% (25 of 34) of LT, and 67% (20 of 30) of LTST ETEC. When colony lysates of these ETEC were examined, the AP probes identified all 94 ETEC. In examinations of stool blots, the RL and AP probes were shown to have sensitivities of 71 and 59%, specificities of 91 and 86%, positive predictive values of 87 and 73%, and negative predictive values of 86 and 74%, respectively. AP oligonucleotide probes to detect ETEC were less sensitive in detecting ETEC by colony or stool blot hybridization than the RL probes but could be used by laboratories without access to radioisotopes to examine colony lysates

151

Zinc status and vacuolar zinc transporters control alkaline phosphatase accumulation and activity in Saccharomyces cerevisiae  

Science.gov (United States)

SUMMARY Little is known about how metalloproteins in the secretory pathway obtain their metal ion cofactors. We used the Pho8 alkaline phosphatase of the yeast Saccharomyces cerevisiae to probe this process in vivo. We found that both Pho8 activity and protein accumulation are zinc dependent and decrease in zinc-limited cells. Low Pho8 accumulation was the result of degradation by vacuolar proteases. Surprisingly, the protective effect of zinc on Pho8 stability was not solely due to Zn2+ binding to the active site indicating that the Pho8 protein is targeted for degradation in zinc-limited cells by another mechanism. Pho8 is a rare example of a metalloprotein whose stability is regulated by its metal cofactor independently of active site binding. We also assessed which zinc transporters are responsible for supplying zinc to Pho8. We found that the Zrc1 and Cot1 vacuolar zinc transporters play the major role while the Msc2/Zrg17 zinc transporter complex active in the endoplasmic reticulum is not involved. These results demonstrate that the vacuolar zinc transporters, previously implicated in metal detoxification, also deliver zinc to certain metalloproteins within intracellular compartments. These data suggest that Pho8 receives its metal cofactor in the vacuole rather than in earlier compartments of the secretory pathway. PMID:19298366

Qiao, Wei; Ellis, Charissa; Steffen, Janet; Wu, Chang-Yi; Eide, David J.

2009-01-01

152

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

International Nuclear Information System (INIS)

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

153

Curcumin and Chronic Kidney Disease (CKD: Major Mode of Action through Stimulating Endogenous Intestinal Alkaline Phosphatase  

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Full Text Available Curcumin, an active ingredient in the traditional herbal remedy and dietary spice turmeric (Curcuma longa, has significant anti-inflammatory properties. Chronic kidney disease (CKD, an inflammatory disease, can lead to end stage renal disease resulting in dialysis and transplant. Furthermore, it is frequently associated with other inflammatory disease such as diabetes and cardiovascular disorders. This review will focus on the clinically relevant inflammatory molecules that play a role in CKD and associated diseases. Various enzymes, transcription factors, growth factors modulate production and action of inflammatory molecules; curcumin can blunt the generation and action of these inflammatory molecules and ameliorate CKD as well as associated inflammatory disorders. Recent studies have shown that increased intestinal permeability results in the leakage of pro-inflammatory molecules (cytokines and lipopolysaccharides from gut into the circulation in diseases such as CKD, diabetes and atherosclerosis. This change in intestinal permeability is due to decreased expression of tight junction proteins and intestinal alkaline phosphatase (IAP. Curcumin increases the expression of IAP and tight junction proteins and corrects gut permeability. This action reduces the levels of circulatory inflammatory biomolecules. This effect of curcumin on intestine can explain why, despite poor bioavailability, curcumin has potential anti-inflammatory effects in vivo and beneficial effects on CKD.

Siddhartha S. Ghosh

2014-12-01

154

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

Science.gov (United States)

Titania nanotubes (TNTs) were prepared by anodization on different substrates (titanium, Ti6Al4V and Ti6Al7Nb alloys) in ethylene glycol and glycerol. The influence of the applied potential and processing time on the nanotube diameter and length is analyzed. The as-formed nanotube layers are amorphous but they become crystalline when subjected to subsequent thermal treatment in air at 550°C; TNT layers grown on titanium and Ti6Al4V alloy substrates consist of anatase and rutile, while those grown on Ti6Al7Nb alloy consist only of anatase. The nanotube layers grown on Ti6Al7Nb alloy are less homogeneous, with supplementary islands of smaller diameter nanotubes, spread across the surface. Better adhesion and proliferation of osteoblasts was found for the nanotubes grown on all three substrates by comparison to an unprocessed titanium plate. The sensitivity towards bovine alkaline phosphatase was investigated mainly by electrochemical impedance spectroscopy in relation to the crystallinity, the diameter and the nature of the anodization electrolyte of the TNT/Ti samples. The measuring capacity of the annealed nanotubes of 50nm diameter grown in glycerol was demonstrated and the corresponding calibration curve was built for the concentration range of 0.005-0.1mg/mL. PMID:24582263

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

2014-04-01

155

A folding study of Antarctic krill (Euphausia superba) alkaline phosphatase using denaturants.  

Science.gov (United States)

To gain insight into the structural and folding mechanisms of Antarctic krill alkaline phosphatase (ALP), the enzyme was properly purified by (NH4)2SO4 fractionation and by both Sephadex G-75 and DEAE anion exchange chromatography. The purified enzyme (62.6 kDa; 2.62 unit/mg) was unstable at temperatures exceeding 30°C. Denaturants, such as sodium dodecyl sulfate (SDS), guanidine HCl, and urea, were applied to evaluate the folding mechanism, including kinetics and thermodynamics, of krill ALP. Sodium dodecyl sulfate elicited no significant effect on ALP activity even at excessively high concentrations (300 mM), whereas guanidine HCl and urea effectively inactivated the enzyme at concentrations of 2 and 3.5 M, respectively. Kinetic studies showed that the enzymatic inhibition by guanidine HCl and urea represented a first-order reaction that was a monophasic unfolding process. This process was found to be associated with conformational changes without significant transient free-energy changes. Additionally, the overall structural changes occurred proximally to the active site pocket. Our study provides new insight into ALP of the Antarctic krill, which lives in extreme environmental conditions. PMID:25016161

Wang, Zhi-Jiang; Lee, Jinhyuk; Si, Yue-Xiu; Wang, Wei; Yang, Jun-Mo; Yin, Shang-Jun; Qian, Guo-Ying; Park, Yong-Doo

2014-09-01

156

In Vivo Overexpression of Tissue-Nonspecific Alkaline Phosphatase Increases Skeletal Mineralization and Affects the Phosphorylation Status of Osteopontin  

OpenAIRE

Functional ablation of tissue-nonspecific alkaline phosphatase (TNAP) (Alpl?/? mice) leads to hypophosphatasia, characterized by rickets/osteomalacia attributable to elevated levels of extracellular inorganic pyrophosphate, a potent mineralization inhibitor. Osteopontin (OPN) is also elevated in the plasma and skeleton of Alpl?/? mice. Phosphorylated OPN is known to inhibit mineralization, however, the phosphorylation status of the increased OPN found in Alpl?/? mice is unknown. H...

Narisawa, Sonoko; Yadav, Manisha C.; Milla?n, Jose? Luis

2013-01-01

157

Effect of endosulfan on acid and alkaline phosphatase activity in liver, kidney, and muscles of Channa gachua  

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The widespread use of a great many toxic chemicals to eliminate unwanted plant or animal species has resulted in the contamination of most aquatic habitats with these substances on a regular basis. Endosulfan, a polycyclic chlorinated hydrocarbon of cyclodien group, is a well known organochlorine insecticide on the activity of acid and alkaline phosphatase in liver, kidney and muscles of a freshwater teleost, Channa gachua.

Sharma, R.M. (Jiwaji Univ., Gwalior (India))

1990-03-01

158

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

OpenAIRE

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 plasma cell membrane glycoprotein-1 (PC-1) are involved in this process. Mutations in the TNAP gene result in the inborn error of metabolism known as hypophosphatasia, charact...

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

2002-01-01

159

Interactions between CD36 and global intestinal alkaline phosphatase in mouse small intestine and effects of high-fat diet  

OpenAIRE

The mechanisms of the saturable component of long-chain fatty acid (LCFA) transport across the small intestinal epithelium and its regulation by a high-fat diet (HFD) are uncertain. It is hypothesized here that the putative fatty acid translocase/CD36 and intestinal alkaline phosphatases (IAPs) function together to optimize LCFA transport. Phosphorylated CD36 (pCD36) was expressed in mouse enterocytes and dephosphorylated by calf IAP (CIAP). Uptake of fluorescently tagged LCFA into isolated e...

Lynes, Matthew; Narisawa, Sonoko; Milla?n, Jose? Luis; Widmaier, Eric P.

2011-01-01

160

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

OpenAIRE

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 resi...

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.

2006-01-01

161

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

OpenAIRE

We measured pools of dissolved phosphorus (P), including dissolved inorganic P (DIP), dissolved organic P (DOP) and alkaline phosphatase (AP)-hydrolyzable labile DOP (L-DOP), and kinetic parameters of AP activity (APA) in the euphotic zone in the western North Pacific Ocean. Samples were collected from one coastal station in Sagami Bay, Japan, and three offshore stations between the North Pacific subtropical gyre (NPSG) and the Kuroshio region. Although DIP concentrations in the euphotic zone...

MasahiroSuzumura; FuminoriHashihama

2012-01-01

162

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

OpenAIRE

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

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

2011-01-01

163

The Effects of Culture Conditions on the Glycosylation of Secreted Human Placental Alkaline Phosphatase Produced in Chinese Hamster Ovary Cells  

OpenAIRE

The effects of different culture conditions, suspension and microcarrier culture and temperature reduction on the structures of N-linked glycans attached to secreted human placental alkaline phosphatase (SEAP) were investigated for CHO cells grown in a controlled bioreactor. Both mass spectrometry and anion-exchange chromatography were used to probe the N-linked glycan structures and distribution. Complex-type glycans were the dominant structures with small amounts of high mannose glycans obs...

Nam, Jong Hyun; Zhang, Fuming; Ermonval, Myriam; Linhardt, Robert J.; Sharfstein, Susan T.

2008-01-01

164

Reduced Levels of Membrane-Bound Alkaline Phosphatase Are Common to Lepidopteran Strains Resistant to Cry Toxins from Bacillus thuringiensis  

OpenAIRE

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 Heliot...

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

2011-01-01

165

Hormonal regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase and alkaline phosphatase in cultured intestinal mucosa.  

Science.gov (United States)

The endocrine regulation of the key enzyme of cholesterol synthesis, 3-hydroxy-3-methylglutaryl-CoA reductase (EC 1.1.1.34) and of the brush border enzyme alkaline phosphatase (EC 3.1.3.1) was studied in short (2 h) and long term (24 h) organ culture of rabbit ileum mucosa. In contrast to the hepatic enzyme, intestinal reductase is not subject to regulation by insulin or glucagon even at a pharmacological level. This applies to both 'total' and 'active' reductase, prepared in the absence or presence of sodium fluoride, respectively. During culture, there is a gradual, time-dependent increase in the active, dephosphorylated enzyme form. This endogenous activation was found to be unaffected by all hormones tested. Similarly, alkaline phosphatase was not influenced by both pancreatic hormones. In contrast, triamcinolone significantly (P less than 0.05) suppressed reductase in a dose-dependent fashion to 38% of controls after 24 h, but not after 2 h culture. Alkaline phosphatase was induced after both periods, but the effect was more marked after 24 h. A parallel minor stimulation of both enzyme activities was noted in the presence of 10(-9)M triiodothyronine (P less than 0.05), lower and very high (10(-5)M) concentrations were ineffective. In view of the role of glucocorticoids as intestinal growth inhibitors and of thyroid hormones as growth stimulators, it is suggested that changes in reductase reflect alterations of crypt membrane cholesterol synthesis, whereas the induction of alkaline phosphatase is mediated through an enhanced enterocyte regeneration and/or maturation. PMID:7032602

Stange, E F; Preclik, G; Schneider, A; Seiffer, E; Ditschunneit, H

1981-12-01

166

Characterization of alkaline phosphatase labeled UidA(Gus) probe and its application in testing of transgenic tritordeum.  

Science.gov (United States)

Hybridization is a very important molecular biology technique to measure the degree of genetic similarity between DNA sequences, and detect the foreign genes in transgenic organisms. To label a DNA or RNA probe plays a key role in hybridization. A method using nonradioactive material alkaline phosphatase to label UidA(Gus) DNA as probe has been studied. On that basis of Renz and our previous work, alkaline phosphatase-labeled DNA was used as a probe to examine the transformation of the foreign UidA(Gus) gene in transgenic tritordeum. Such DNA-enzyme complexes were characterized and examined carefully, the results showed that it was a sensitive, specific, safe and economical probe. For dot hybridization and Southern blot under full-stringency conditions with alkaline phosphatase as the detector and 5-bromo-4-chloro-3-indolyl phosphate (BCIP)-Nitro Blue Tetrazolium (NBT) as the substrate, dot hybridization showed that the UidA(Gus) gene was transformed into the target plants and inherited stable, Southern blot showed that at least two copies of UidA(Gus) gene were inserted into one line of our transgenic tritordeum. Histochemical staining with X-Gluc of transgenic tritordeum also certified that the foreign UidA(Gus) DNA were transformed into the transgenic tritordeum. PMID:21153926

Tu, Zhiming; Zhang, Jiangzhou; Yang, Guangxiao; He, Guangyuan

2011-08-01

167

A novel bifunctional hybrid with marine bacterium alkaline phosphatase and Far Eastern holothurian mannan-binding lectin activities.  

Science.gov (United States)

A fusion between the genes encoding the marine bacterium Cobetia marina alkaline phosphatase (CmAP) and Far Eastern holothurian Apostichopus japonicus mannan-binding C-type lectin (MBL-AJ) was performed. Expression of the fusion gene in E. coli cells resulted in yield of soluble recombinant chimeric protein CmAP/MBL-AJ with the high alkaline phosphatase activity and specificity of the lectin MBL-AJ. The bifunctional hybrid CmAP/MBL-AJ was produced as a dimer with the molecular mass of 200 kDa. The CmAP/MBL-AJ dimer model showed the two-subunit lectin part that is associated with two molecules of alkaline phosphatase functioning independently from each other. The highly active CmAP label genetically linked to MBL-AJ has advantaged the lectin-binding assay in its sensitivity and time. The double substitution A156N/F159K in the lectin domain of CmAP/MBL-AJ has enhanced its lectin activity by 25 ± 5%. The bifunctional hybrid holothurian's lectin could be promising tool for developing non-invasive methods for biological markers assessment, particularly for improving the MBL-AJ-based method for early detection of a malignant condition in cervical specimens. PMID:25397876

Balabanova, Larissa; Golotin, Vasily; Kovalchuk, Svetlana; Bulgakov, Alexander; Likhatskaya, Galina; Son, Oksana; Rasskazov, Valery

2014-01-01

168

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

OpenAIRE

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 pro...

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

2002-01-01

169

Loss of Skeletal Mineralization by the Simultaneous Ablation of PHOSPHO1 and Alkaline Phosphatase Function: A Unified Model of the Mechanisms of Initiation of Skeletal Calcification  

OpenAIRE

Endochondral ossification is a carefully orchestrated process mediated by promoters and inhibitors of mineralization. Phosphatases are implicated, but their identities and functions remain unclear. Alkaline phosphatase (TNAP) plays a crucial role promoting mineralization of the extracellular matrix by restricting the concentration of the calcification inhibitor inorganic pyrophosphate (PPi). Mutations in the TNAP gene cause hypophosphatasia, a heritable form of rickets and osteomalacia. Here ...

Yadav, Manisha C.; Sima?o, Ana Maria Sper; Narisawa, Sonoko; Huesa, Carmen; Mckee, Marc D.; Farquharson, Colin; Milla?n, Jose? Luis

2010-01-01

170

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

International Nuclear Information System (INIS)

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

171

Effects of vasectomy on seminal plasma alkaline phosphatase in male alpacas (Vicugña pacos).  

Science.gov (United States)

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. PMID:23790090

Pearson, L K; Campbell, A J; Sandoval, S; Tibary, A

2013-12-01

172

Effects of amines and aminoalcohols on bovine intestine alkaline phosphatase activity.  

Science.gov (United States)

Bovine intestine alkaline phosphatase (BIALP) is widely used as a signaling enzyme in sensitive assays such as enzyme immunoassay (EIA). In this study, we evaluated the effects of various aminoalcohols and amines on the activity of BIALP in the hydrolysis of p-nitrophenyl phosphate (pNPP) at pH 9.8, at 20 °C. The k(cat) values at 0.05 M diethanolamine, 0.1 M triethanolamine, and 0.2 M N-methylethanolamine were 190±10, 840±30, and 500±10 s(-1), respectively. The k(cat) values increased with increasing concentrations of diethanolamine, triethanolamine, and N-methylethanolamine and reached 1240±60, 1450±30, and 2250±80 s(-1), respectively, at 1.0M. On the other hand, the k(cat) values at 0.05-1.0M ethanolamine, ethylamine, methylamine, and dimethylamine were in the range of 100-600 s(-1). These results indicate that diethanolamine, triethanolamine and N-methylethanolamine highly activate BIALP and might be suitable as a dilution buffer of BIALP in EIA. Interestingly, the K(m) values increased with increasing concentrations of diethanolamine and N-methylethanolamine, but not triethanolamine: the K(m) value at 1.0M diethanolamine (0.83±0.15 mM) was 12-fold higher than that at 0.05M (0.07±0.01 mM), and that at 1.0M N-methylethanolamine (2.53±0.20 mM) was 14-fold higher than that at 0.2M (0.18±0.02 mM), while that at 1.0M triethanolamine (0.31±0.01 mM) was similar as that at 0.2M (0.25±0.01 mM), suggesting that the mechanisms of BIALP activation are different between the aminoalcohols. PMID:22112405

Sekiguchi, Satoshi; Hashida, Yasuhiko; Yasukawa, Kiyoshi; Inouye, Kuniyo

2011-07-10

173

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English 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 30 [...] ºC. 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.

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

2000-08-01

174

Pathophysiological role of vascular smooth muscle alkaline phosphatase in medial artery calcification.  

Science.gov (United States)

Medial vascular calcification (MVC) is a pathological phenomenon that causes vascular stiffening and can lead to heart failure; it is common to a variety of conditions, including aging, chronic kidney disease, diabetes, obesity, and a variety of rare genetic diseases. These conditions share the common feature of tissue-nonspecific alkaline phosphatase (TNAP) upregulation in the vasculature. To evaluate the role of TNAP in MVC, we developed a mouse model that overexpresses human TNAP in vascular smooth muscle cells in an X-linked manner. Hemizygous overexpressor male mice (Tagln-Cre(+/-) ; Hprt(ALPL) (/Y) or TNAP-OE) show extensive vascular calcification, high blood pressure, and cardiac hypertrophy, and have a median age of death of 44 days, whereas the cardiovascular phenotype is much less pronounced and life expectancy is longer in heterozygous (Tagln-Cre(+/-) ; Hprt(ALPL) (/-) ) female TNAP-OE mice. Gene expression analysis showed upregulation of osteoblast and chondrocyte markers and decreased expression of vascular smooth muscle markers in the aortas of TNAP-OE mice. Through medicinal chemistry efforts, we developed inhibitors of TNAP with drug-like pharmacokinetic characteristics. TNAP-OE mice were treated with the prototypical TNAP inhibitor SBI-425 or vehicle to evaluate the feasibility of TNAP inhibition in vivo. Treatment with this inhibitor significantly reduced aortic calcification and cardiac hypertrophy, and extended lifespan over vehicle-treated controls, in the absence of secondary effects on the skeleton. This study shows that TNAP in the vasculature contributes to the pathology of MVC and that it is a druggable target. © 2015 American Society for Bone and Mineral Research. PMID:25428889

Sheen, Campbell R; Kuss, Pia; Narisawa, Sonoko; Yadav, Manisha C; Nigro, Jessica; Wang, Wei; Chhea, T Nicole; Sergienko, Eduard A; Kapoor, Kapil; Jackson, Michael R; Hoylaerts, Marc F; Pinkerton, Anthony B; O'Neill, W Charles; Millán, José Luis

2015-05-01

175

Recombinant Production and Characterization of a Highly Active Alkaline Phosphatase from Marine Bacterium Cobetia marina.  

Science.gov (United States)

The psychrophilic marine bacterium, Cobetia marina, recovered from the mantle tissue of the marine mussel, Crenomytilus grayanus, which contained a gene encoding alkaline phosphatase (AP) with apparent biotechnology advantages. The enzyme was found to be more efficient than its counterparts and showed k cat value 10- to 100-fold higher than those of all known commercial APs. The enzyme did not require the presence of exogenous divalent cations and dimeric state of its molecule for activity. The recombinant enzyme (CmAP) production and purification were optimized with a final recovery of 2 mg of the homogenous protein from 1 L of the transgenic Escherichia coli Rosetta(DE3)/Pho40 cells culture. CmAP displayed a half-life of 16 min at 45 °C and 27 min at 40 °C in the presence of 2 mM EDTA, thus suggesting its relative thermostability in comparison with the known cold-adapted analogues. A high concentration of EDTA in the incubation mixture did not appreciably inhibit CmAP. The enzyme was stable in a wide range of pH (6.0-11.0). CmAP exhibited its highest activity at the reaction temperature of 40-50 °C and pH 9.5-10.3. The structural features of CmAP could be the reason for the increase in its stability and catalytic turnover. We have modeled the CmAP 3D structure on the base of the high-quality experimental structure of the close homologue Vibrio sp. AP (VAP) and mutated essential residues predicted to break Mg(2+) bonds in CmAP. It seems probable that the intrinsically tight binding of catalytic and structural metal ions together with the flexibility of intermolecular and intramolecular links in CmAP could be attributed to the adapted mutualistic lifestyle in oceanic waters. PMID:25260971

Golotin, Vasily; Balabanova, Larissa; Likhatskaya, Galina; Rasskazov, Valery

2015-04-01

176

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

Directory of Open Access Journals (Sweden)

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

2012-12-01

177

Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity  

Scientific Electronic Library Online (English)

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

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

2008-07-01

178

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

2013-10-15

179

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

Science.gov (United States)

Alkaline phosphatase (AP) from the moderate halophilic bacterium Halomonas sp. 593 (HaAP) catalyzes the hydrolysis of phosphomonoesters over a wide salt-concentration range (1-4?M NaCl). In order to clarify the structural basis of its halophilic characteristics and its wide-range adaptation to salt concentration, the tertiary structure of HaAP was determined by X-ray crystallography to 2.1?Å resolution. The unit cell of HaAP contained one dimer unit corresponding to the biological unit. The monomer structure of HaAP contains a domain comprised of an 11-stranded ?-sheet core with 19 surrounding ?-helices similar to those of APs from other species, and a unique `crown' domain containing an extended `arm' structure that participates in formation of a hydrophobic cluster at the entrance to the substrate-binding site. The HaAP structure also displays a unique distribution of negatively charged residues and hydrophobic residues in comparison to other known AP structures. AP from Vibrio sp. G15-21 (VAP; a slight halophile) has the highest similarity in sequence (70.0% identity) and structure (C(?) r.m.s.d. of 0.82?Å for the monomer) to HaAP. The surface of the HaAP dimer is substantially more acidic than that of the VAP dimer (144 exposed Asp/Glu residues versus 114, respectively), and thus may enable the solubility of HaAP under high-salt conditions. Conversely, the monomer unit of HaAP formed a substantially larger hydrophobic interior comprising 329 C atoms from completely buried residues, whereas that of VAP comprised 264 C atoms, which may maintain the stability of HaAP under low-salt conditions. These characteristics of HaAP may be responsible for its unique functional adaptation permitting activity over a wide range of salt concentrations. PMID:24598750

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

2014-03-01

180

Contribution of matrix vesicles and alkaline phosphatase to ectopic bone formation  

Scientific Electronic Library Online (English)

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

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

2006-05-01

181

Characterization of a serum factor interfering with the radioimmunoassay of prostatic acid phosphatase  

International Nuclear Information System (INIS)

Storage of serum at temperatures above zero for short periods results in the spurious formation of a factor interfering with the radioimmunoassay of prostatic acid phosphatase. This factor acts by inhibiting the binding of radiolabelled tracer to the specific primary antiserum. The inhibiting factor is a protein of molecular mass 40 000 which is derived from an inactive high molecular mass precursor. Formation of the inhibiting factor is apparently reversible and its action can be neutralized by adding fresh serum to the incubation mixture. (Auth.)

182

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

Science.gov (United States)

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

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

2013-10-01

183

Influence of zinc ions addition to different lots of 2-amino-2-methyl-1-propanol (AMP) buffer on the alkaline phosphatase activities.  

Science.gov (United States)

The influence of Zn2+ on the inactivation of alkaline phosphatase [orthophosphoric monoester phosphohydrolase (alkaline optimum), EC 3.1 3.1] in serum during preincubation with 2-amino-2-methyl-1-propanol (AMP) buffers was investigated. Addition of Zn2+ to the buffer before preincubation increases the enzyme activity. An optimum Zn2+ concentration different for each lot of AMP buffer can be found, at which the enzyme activities are restored to a level equal to activities measured without preincubation. There is a relation between the inactivating properties of the different AMP buffers and the amount of Zn2+ needed to prevent this inactivation. Since Zn2+ chelating substituted diamines are held responsible for the inactivation by removing Zn2+ from the enzyme, we assume that the addition of Zn2+ to the buffer prevents this removal. As Zn2+ itself is an inhibitor of the enzyme, the addition of both too much or too little Zn2+ results in lower enzyme activities after preincubation with AMP buffer. PMID:498532

Pekelharing, J M; Noordeloos, P J; Leijnse, B

1979-10-15

184

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

Directory of Open Access Journals (Sweden)

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

Bednyakov Dmitriy Andreevich

2012-11-01

185

Toxic impact of aldrin on acid and alkaline phosphatase activity of penaeid prawn, Metapenaeus monoceros: In vitro study  

Energy Technology Data Exchange (ETDEWEB)

The increasing contamination of the aquatic environment by the indiscriminate and widespread use of different kinds of pesticides is a serious problem for environmental biologists. Organochlorine insecticides are more hazardous since they are not only more toxic but also leave residues in nature. The deleterious effects of aldrin on several crustaceans have been studied. But studies concerning the impact of aldrin on biochemical aspects of crustaceans are very much limited. The present study is aimed at probing the in vitro effects of aldrin on the acid and alkaline phosphatase activity levels in selected tissues of penaeid prawn, Metapenaeus monoceros (Fabricius).

Reddy, M.S.; Jayaprada, P.; Rao, K.V.R. (Sri Venkateswara Univ. Post Graduate Center, Kavali (India))

1991-03-01

186

Aldrin and lindane impact on acid and alkaline phosphatase activities of prawn, Metapenaeus monoceros: in vitro study.  

Science.gov (United States)

Activity levels of acid and alkaline phosphatases in hepatopancreas, stomach, muscle, gill and brain tissues of penaeid prawn, Metapenaeus monoceros were studied after in vitro addition of different concentrations of aldrin and lindane, the organochlorine insecticides. The activity levels of both these enzymes were inhibited significantly in all the tissues of prawn and the degree of inhibition is increased with increase in the concentration of insecticide. The significant inhibition creates disturbances in the normal functioning such as disturbances in the normal functioning such as disturbances in protein synthesis and different biochemical lesions in selected tissues of prawn, M. monoceros. PMID:1708667

Reddy, M S; Rao, K V

1990-12-01

187

Markedly increased circulating pyridoxal-5'-phosphate levels in hypophosphatasia. Alkaline phosphatase acts in vitamin B6 metabolism.  

OpenAIRE

Markedly increased circulating concentrations of pyridoxal-5'-phosphate (PLP) were found in each of 14 patients representing all clinical forms of hypophosphatasia, an inborn error characterized by deficient activity of the tissue nonspecific (bone/liver/kidney) isoenzyme of alkaline phosphatase (AP). The mean PLP concentration in plasma was 1174 nM (range, 214-3839 nM) in the patients and 57 +/- 26 nM (mean +/- SD) in 38 control subjects. In four affected children, urinary excretion of the P...

Whyte, M. P.; Mahuren, J. D.; Vrabel, L. A.; Coburn, S. P.

1985-01-01

188

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

189

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

Science.gov (United States)

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 complex 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. PMID:9285892

Barrio, D A; Braziunas, M D; Etcheverry, S B; Cortizo, A M

1997-06-01

190

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

Science.gov (United States)

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. PMID:22939148

Santos, Inês C; Mesquita, Raquel B R; Bordalo, Adriano A; Rangel, António O S S

2012-08-30

191

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

192

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

International Nuclear Information System (INIS)

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

193

Multiple forms of human intestinal alkaline phosphatase: chemical and enzymatic properties, and circulating clearances of the fast- and slow-moving enzymes  

International Nuclear Information System (INIS)

Two forms of alkaline phosphatase orthophosphoric monoester phosphohydrolase (alkaline optimum, EC 3.1.3.1) have been purified from human small intestine by column chromatography on DEAE-cellulose and tyraminyl derivative affinity gel, and by preparative disc gel electrophoresis. Intestinal phosphatases were electrophoretically separated into two components, fast- and slow-moving enzymes, with apparent molecular weights of 140000 and 168000 and with subunit weights of 68000 and 80000, respectively. Organ distribution of injected 125I-labelled enzymes indicates that the desialylated hepatic enzyme was selectively distributed in liver, while the degalactosylated intestinal enzyme was incorporated into liver, lymph fluid, and small intestine. These results suggest that the pathway of circulating clearance of alkaline phosphatase has several routes. (Auth.)

194

Radioimmunological determination of the prostatic acid phosphatase concentrations in the blood serum of healthy males  

International Nuclear Information System (INIS)

A highly specific and sensitive radioimmunological method is used for determination of the normal blood serum prostatic acid phosphatase (PAP) concentrations in 143 healthy males aged from 21 to 80 years distributed in 6 age groups. The results varied from 2 to 25 pmol/l. A statistically reliable increase (p < 0,025) in PAP-values was detected in males after 60 years of age. The reference values of the PAP-concentrations were elaborated according to the age, which could be used for sure results interpretation in patients with prostate diseases. 1 tab., 1 fig., 4 refs

195

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

Science.gov (United States)

A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode interface, affording fast and highly sensitive ECL cytosensing and cell surface glycan evaluation. Combining the multivalent aptamer interface and ALP nanoprobes, the ECL cytosensor showed a detection limit of 38 CCRF-CEM cells per mL in human serum samples, broad dynamic range and excellent selectivity. In addition, the proposed biosensor provided a valuable insight into dynamic profiling of the expression of different glycans on cell surfaces, based on the carbohydrates recognized by lectins applied to the nanoprobes. This biosensor exhibits great promise in clinical diagnosis and drug screening.A multivalent recognition and alkaline phosphatase (ALP)-responsive electrogenerated chemiluminescence (ECL) biosensor for cancer cell detection and in situ evaluation of cell surface glycan expression was developed on a poly(amidoamine) (PAMAM) dendrimer-conjugated, chemically reduced graphene oxide (rGO) electrode interface. In this strategy, the multivalency and high affinity of the cell-targeted aptamers on rGO provided a highly efficient cell recognition platform on the electrode. The ALP and concanavalin A (Con A) coated gold nanoparticles (Au NPs) nanoprobes allowed the ALP enzyme-catalyzed production of phenols that inhibited the ECL reaction of Ru(bpy)32+ on the rGO electrode interface, affording fast and highly sensitive ECL cytosensing and cell surface glycan evaluation. Combining the multivalent aptamer interface and ALP nanoprobes, the ECL cytosensor showed a detection limit of 38 CCRF-CEM cells per mL in human serum samples, broad dynamic range and excellent selectivity. In addition, the proposed biosensor provided a valuable insight into dynamic profiling of the expression of different glycans on cell surfaces, based on the carbohydrates recognized by lectins applied to the nanoprobes. This biosensor exhibits great promise in clinical diagnosis and drug screening. Electronic supplementary information (ESI) available: CV and EIS during the electrode assembly, activity of the nanoprobes and the glycan-binding specificities of the lectins. See DOI: 10.1039/c4nr03053b

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

2014-09-01

196

Prognostic role of serum prostatic acid phosphatase for 103Pd-based radiation for prostatic carcinoma  

International Nuclear Information System (INIS)

Purpose: To establish the prognostic role of serum enzymatic prostatic acid phosphatase (PAP) in patients treated with palladium (103Pd) and supplemental external beam irradiation (EBRT) for clinically localized, high-risk prostate carcinoma. Methods and Materials: One hundred twenty-four consecutive patients with Stage T2a-T3 prostatic carcinoma were treated from 1992 through 1995. Each patient had at least one of the following risk factors for extracapsular disease extension: Stage T2b or greater (100 patients), Gleason score 7-10 (40 patients), pretreatment prostate specific antigen (PSA) > 15 ng/ml (32 patients), or elevated serum PAP (25 patients). Patients received 41 Gy conformal EBRT to a limited pelvic field, followed 4 weeks later by a 103Pd boost (prescription dose 80 Gy). Biochemical failure was defined as a PSA greater than 1 ng/ml (normal < 4 ng/ml). Results: The overall, actuarial freedom from biochemical failure at 4 years after treatment was 79%. In Cox-proportional hazard multivariate analysis, the strongest predictor of failure was elevated pretreatment acid phosphatase (p = 0.02), followed by Gleason score (p = 0.1), and PSA (p = 0.14). Conclusion: PAP was the strongest predictor of long-term biochemical failure. It may be a more accurate indicator of micrometastatic disease than PSA, and as such, we suggest that it be reconsidered for general use in radiation-treated patients

197

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

Science.gov (United States)

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. PMID:45242

Carpenè, E; Crisetig, G; Cortesi, P; Serrazanetti, G

1979-07-15

198

Dicistronic LacZ and alkaline phosphatase reporter constructs permit simultaneous histological analysis of expression from multiple transgenes.  

Science.gov (United States)

We report here the development of convenient dicistronic transgenic markers for the rapid and efficient simultaneous analysis of transgene activity in transgenic mice. Two sensitive histological markers, the beta-galactosidase (beta-gal)-encoding lacZ gene and the human placental alkaline phosphatase (hpAP) gene, have been fused to the internal ribosome entry sequence (IRES) from the encephalomyocarditis virus, which directs efficient mRNA cap-independent entry of the translation apparatus in mammalian cells. The IRES permits efficient translation of either lacZ or hpAP when placed anywhere within transgene exonic sequences, including both 5' and 3' untranslated regions. In addition, the production of constructs for transgenic analysis of DNA regulatory elements is greatly facilitated with IRES-lacZ or IRES-hpAP, since the IRES relieves the need for complicated in frame transgene protein fusions to produce a functional beta-gal or hpAP protein. PMID:9383553

Li, X; Wang, W; Lufkin, T

1997-11-01

199

Radionuclide bone scan, radiographic bone survey, and alkaline phosphatase: studies of limited value in asymptomatic patients with ovarian carcinoma  

International Nuclear Information System (INIS)

Bone scans or skeletal surveys were obtained in 104 patients with ovarian carcinoma. No metastases were identified at staging in the 43 patients with Stage I or II disease. Four patients in the entire series had osseous metastases. Three of the 40 patients with Stage III epithelian ovarian carcinoma has osseous metastases at the time of staging. All of these were Grade III lesions. One Stage I, Grade III patient demonstrated osseous metastases two years after initial diagnosis. None of the four patients with osseous metastases had an elevated alkaline phosphatase; three of the four had bone pain. Based on these results, it is suggested that radiographic bone survey and radionuclide bone scans are not indicated as screening procedures in asymptomatic patients with ovarian carcinoma

200

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

Science.gov (United States)

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

Faure, Mathilde; Sotta, Bruno; Gamby, Jean

2014-08-15

201

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

202

Real-time fluorescence assays of alkaline phosphatase and ATP sulfurylase activities based on a novel PPi fluorescent probe.  

Science.gov (United States)

An anthracene-armed tetraaza macrocyclic fluorescent probe 3-(9-anthrylmethyl)-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene(l) for detecting Zn(2+) in aqueous medium was synthesized. L-Zn(2+) complex, showed selectivity toward pyrophosphate ion (PPi) by quenching the fluorescence in aqueous HEPES buffer (pH 7.4). Furthermore, L-Zn(2+) was also used to set up a real-time fluorescence assay for monitoring enzyme activities of alkaline phosphatase (ALP) and adenosine triphosphate sulfurylase (ATPS). In the presence of ALP inhibitor Na3VO4 and ATPS inhibitor chlorate, two enzymes activities decreased obviously, respectively. PMID:25770619

Wang, Xiaobo; Zhang, Zhiyang; Ma, Xiaoyan; Wen, Jinghan; Geng, Zhirong; Wang, Zhilin

2015-05-01

203

Activities and distribution of alkaline phosphatase and carbonic anhydrase in the tibial dyschondroplastic lesion and associated growth plate of chicks.  

Science.gov (United States)

The tibial dyschondroplastic lesion and associated growth plate were examined for the presence of alkaline phosphatase, an enzyme important in cartilage mineralization, and carbonic anhydrase, an enzyme that regulates acid-base balance of many tissues including cartilage. By histochemistry, both enzymes were found to be present in the prehypertrophic and hypertrophic zones of the growth plate as occurs normally. Enzyme activity was normal or close to normal in the growth plate. Activity was diminished to half of the normal values in the proximal lesion and to zero in the distal lesion. The enzymes are believed to be diminished by necrotic and autolytic processes and are not involved in initiating the formation of the lesion. PMID:3935100

Gay, C V; Anderson, R E; Leach, R M

1985-01-01

204

ALKALINE PHOSPHATASE REACTIVITY IN THE VAGINA AND UTEROVAGINAL JUNCTION SPERM-STORAGE TUBULES OF TURKEYS IN EGG PRODUCTION: IMPLICATIONS FOR SPERM STORAGE  

Science.gov (United States)

Currently there remains contradictory information on the localization and possible role of alkaline phosphatase (AP) in the chicken and Japanese quail oviducts. Using turkeys with a hard-shelled egg in their uteri, vaginal and uterovaginal junction mucosae were stretched and fixed as whole mounts ...

205

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

Science.gov (United States)

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

206

Microheterogeneity of the hydrophobic and hydrophilic part of the glycosylphosphatidylinositol anchor of alkaline phosphatase from calf intestine.  

Science.gov (United States)

Digestion of calf intestine alkaline phosphatase with pronase and subsequent dephosphorylation of the released peptidyl-(Etn-P)2-glycosyl-PtdIns with HF generated 8 glycosyl-Ins species the largest of which (G1 and G2) have the following proposed structures: [sequence: see text] G3 and G5 are lower homologues of G1 and G2, respectively, being one alpha 1-2 linked mannopyranosyl residue shorter. G4 is analogous to G2 lacking the N-acetylgalactosaminyl residue and G6 is the next lower homologue of G4. Most of G4 and G6 occur substituted with a palmitoyl (G4, G6) or a myristoyl residue (G6) probably attached to the inositol moiety. Thus, the basic ManxGlc-Ins species are either substituted with an N-acetylgalactosaminyl residue or a fatty acid ester. The structures were deduced from compositional analysis, molecular-mass determination by matrix-assisted laser desorption MS, sequential hydrolysis with appropriate exoglycosidases and treatment with CrO3. Purification of the glycosylinositol species was achieved by a novel reverse-phase HPLC technique using fluorescent fluoren-9-yl-methoxy-carbonyl (Fmoc) derivatives. These stable derivatives were susceptible to hydrolysis with exoglycosidases which allowed sequential cleavages to be carried out and kinetics to be followed at the picomole level. We observed recently that native alkaline phosphatase separates on octyl-Sepharose into four distinct fractions of increasing hydrophobicity (F1-F4). Here we show that all four fractions contain G1-G6. The acylated species G4 and G6 were restricted to F2 and F4 which had been shown earlier to contain, on average, 2.5 and 3 fatty acid residues/subunit, respectively. In all four fractions the diradylglycerol moiety was predominantly diacylglycerol, alkylacylglycerol being less than 10% which is in contrast to most glycosyl-PtdIns--anchored proteins of mammalian origin. PMID:8665945

Armesto, J; Hannappel, E; Leopold, K; Fischer, W; Bublitz, R; Langer, L; Cumme, G A; Horn, A

1996-05-15

207

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, detachmeated 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

208

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 Dos isoformas de fosfatasas obtenidas de raíz de frijol (Phaseolus vulgaris L.) mostraron un incremento en la actividad en respuesta a la deficiencia de fosfato. Una de ellas (APIII) se purificó a través de una cromatografía de intercambio iónico y una electroforesis preparativa. La masa molecular e [...] stimada para APIII fue de 35 kDa tanto por SDS-PAGE como por filtración molecular, sugiriendo que la enzima activa es monomérica. APIII se clasificó como una fosfatasa alcalina basada en sus requerimientos de pH 8 para catálisis. Esta enzima es activa sobre un amplio espectro de sustratos como polifosfato, glucose 1-fosfato y fosfoenolpiruvato, aunque muestra preferencia por pirofosfato. Su actividad se inhibe completamente por molibdato, vanadato y fosfato, aunque es inhibida parcialmente por fluoruro. Aún cuando los cationes divalentes no fueron escenciales para su actividad, la hidrólisis de pirofosfato se incrementó notablemente en presencia de Mg2+. Abstract in english Two phosphatase isoforms from roots of the common bean (Phaseolus vulgaris L.) showed an increase in activity in response to phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular ma [...] ss of APIII was 35 kDa by both SDS-PAGE and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of pH 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phosphoenol-pyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyrophosphatase activity of this enzyme, the hydrolysis of pyrophosphate increased substantially in the presence of Mg2+.

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

2012-03-01

209

Effect of x-irradiation on the acid and alkaline phosphatase activities in the south Indian scorpion Heterometrus fulvipes (C.L. Koch)  

International Nuclear Information System (INIS)

Variations in the activity levels of acid phosphatase and alkaline phosphatase in the tissues of scorpion Heterometrus fulvipes after whole-body irradiation with X-rays are studied. The animals were exposed to sublethal dose (1/3 of LD50) of radiation for different periods of time ranging from 1/2h to 4h. Tissue homogenates were also irradiated and studied. The activity of phosphatases was found to decrease with increase in exposure time both at in vivo and in vitro, the per cent decrease being higher at in vitro. The activity levels of phosphatases in different tissues were in the order of hepato pancreas > heart pedipalpal > muscle > nervous tissue. (M.G.B.)

210

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

International Nuclear Information System (INIS)

Highlights: • Alkaline phosphatase from eggs of the sea urchin (StAP) proved to be active in seawater. • Activity of StAP is inhibited by very low concentrations of heavy metal. • A test to assess sea and fresh water quality has been developed basing on StAP. • For the first time a salt resistant alkaline phosphatase has been found in eukaryote. - Abstract: A new salt resistant alkaline phosphatase from eggs of the sea urchin Strongylocentrotus intermedius (StAP) has been shown to have a unique property to hydrolyze substrate in seawater without loss of enzymatic activity. The enzyme has pH optimum at 8.0–8.5. Model experiments showed various concentrations of copper, zinc, cadmium and lead added to seawater or a standard buffer mixture to inhibit completely the enzyme activity at the concentrations of 15–150 ?g/l. StAP sensitivity to the presence in seawater of metals, pesticides, detergents and oil products appears to be considerably less. Samples of seawater taken from aquatic areas of the Troitsy Bay of the Peter the Great Bay, Japan Sea have been shown to inhibit the enzyme activity; the same was shown for the samples of fresh waters. The phosphatase inhibition assay developed proved to be highly sensitive, technically easy-to use allowing to test a great number of samples

211

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

International Nuclear Information System (INIS)

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

212

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.

HuiFang Deng

2012-01-01

213

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

International Nuclear Information System (INIS)

Highlights: • Cd2+ reacts with S2? to generate fluorescent CdS QDs with ATP. • ATP can be hydrolyzed by the enzymatic dephosphorylation of ALP. • Fluorescent CdS QDs could not be generated in the presence of ALP. • The analysis system was successfully applied to assay ATP and ALP. - Abstract: We have developed an analytical method to detect adenosine-5?-triphosphate (ATP) and alkaline phosphatase (ALP) based on the generation of CdS quantum dots (QDs). We demonstrated that Cd2+ cation reacts with S2? anion to generate fluorescent CdS QDs in the presence of some certain amount of ATP. With increase in the ATP concentration, the fluorescence intensity of CdS QDs was also enhanced. ATP can be converted into adenosine by the dephosphorylation of ALP, so that the generation of CdS QDs would be inhibited in the presence of ALP. Therefore, this novel analysis system could be applied to assay ATP and ALP based on the growth of fluorescent CdS QDs

214

Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.  

Science.gov (United States)

Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP), Cytokeratin7 (CK7) Cytokeratin8 (CK8) to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinomas, demonstrated CK8 reactivity in 106 (72%) of 148 cases. 95 primary lung carcinoma samples (65%) were positive for CK7. AP was expressed in 81% of primary lung adenocarcinomas. Positive immunoreactivity for AP was characterized by a red, diffusely apical cytoplasmic staining in tumor cells that ocurred singly or in groups. There was a significant difference between AP, CK 7 and CK 8 expressions in primary lung adenocarcinomas (P=0.02; Chi-squared test). The sensitivity of AP, CK8, CK7 as a marker for primary lung adenocarcinomas were 82%, 72%, 64%, respectively. Thus the AP positive staining largely confirmed the cytologic diagnosis of lung adenocarcinoma. PMID:19419944

Serpil, O?uztüzün; Meral, Atay; Müzeyyen, Ozhavzali; Ozlem, Temelli; Umit, Yirtici; Mustafa, Türk; Ziya, Atay

2009-01-01

215

Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.  

Directory of Open Access Journals (Sweden)

Full Text Available Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP, Cytokeratin7 (CK7 Cytokeratin8 (CK8 to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinomas, demonstrated CK8 reactivity in 106 (72% of 148 cases. 95 primary lung carcinoma samples (65% were positive for CK7. AP was expressed in 81% of primary lung adenocarcinomas. Positive immunoreactivity for AP was characterized by a red, diffusely apical cytoplasmic staining in tumor cells that ocurred singly or in groups. There was a significant difference between AP, CK 7 and CK 8 expressions in primary lung adenocarcinomas (P=0.02; Chi-squared test. The sensitivity of AP, CK8, CK7 as a marker for primary lung adenocarcinomas were 82%, 72%, 64%, respectively. Thus the AP positive staining largely confirmed the cytologic diagnosis of lung adenocarcinoma.

Temelli Ozlem

2009-05-01

216

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

Science.gov (United States)

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

217

Valproic Acid Induces Hair Regeneration in Murine Model and Activates Alkaline Phosphatase Activity in Human Dermal Papilla Cells  

Science.gov (United States)

Background Alopecia is the common hair loss problem that can affect many people. However, current therapies for treatment of alopecia are limited by low efficacy and potentially undesirable side effects. We have identified a new function for valproic acid (VPA), a GSK3? inhibitor that activates the Wnt/?-catenin pathway, to promote hair re-growth in vitro and in vivo. Methodology/ Principal Findings Topical application of VPA to male C3H mice critically stimulated hair re-growth and induced terminally differentiated epidermal markers such as filaggrin and loricrin, and the dermal papilla marker alkaline phosphatase (ALP). VPA induced ALP in human dermal papilla cells by up-regulating the Wnt/?-catenin pathway, whereas minoxidil (MNX), a drug commonly used to treat alopecia, did not significantly affect the Wnt/?-catenin pathway. VPA analogs and other GSK3? inhibitors that activate the Wnt/?-catenin pathway such as 4-phenyl butyric acid, LiCl, and BeCl2 also exhibited hair growth-promoting activities in vivo. Importantly, VPA, but not MNX, successfully stimulate hair growth in the wounds of C3H mice. Conclusions/ Significance Our findings indicate that small molecules that activate the Wnt/?-catenin pathway, such as VPA, can potentially be developed as drugs to stimulate hair re-growth. PMID:22506014

Lee, Soung-Hoon; Yoon, Juyong; Shin, Seung Ho; Zahoor, Muhamad; Kim, Hyoung Jun; Park, Phil June; Park, Won-Seok; Min, Do Sik; Kim, Hyun-Yi; Choi, Kang-Yell

2012-01-01

218

Ingestion of potato starch containing esterified phosphorus increases alkaline phosphatase activity in the small intestine in rats.  

Science.gov (United States)

Alkaline phosphatase (ALP) hydrolyzes a variety of monophosphate esters and plays an important role in phosphorus (P) metabolism. Several nutrients in food have been reported to affect intestinal ALP activity in animal models. Previous reports indicated that high levels of P or phosphate in diets decreased intestinal ALP activity in rats. Because potato starch contains considerable amounts of esterified P, unlike other starch-derived plants, we hypothesized that the feeding of potato starch would decrease ALP activity in the intestinal tract. Male Sprague-Dawley rats (7 weeks old) were fed 3 different types of diet containing 60% corn starch or 1 of 2 types of potato starch with different esterified P content for 1 or 5 weeks. Body weight and food intake of each rat were measured every day throughout the experimental periods. At the end of the feeding periods, the small intestine was removed to determine ALP activity in the mucosal tissues. Significant differences were observed in ALP activity in the small intestine between the 2 feeding periods, among the 4 segments of the small intestine, and among the 3 diet groups. Significant positive linear correlations between the amount of P derived from the starch and mucosal ALP activity were obtained in the jejunum and jejunoileum in rats after feeding for 5 weeks. We concluded, contrary to our hypotheses, that the ingestion of potato starch adaptively increases ALP activity in the upper part of the small intestine of growing rats in an esterified P content-dependent manner. PMID:20579526

Mineo, Hitoshi; Morikawa, Nao; Ohmi, Sayako; Ishida, Kyo; Machida, Ayaka; Kanazawa, Takumi; Chiji, Hideyuki; Fukushima, Michihiro; Noda, Takahiro

2010-05-01

219

Increased activity of goat liver plasma membrane alkaline phosphatase upon release by phosphatidylinositol-specific phospholipase C.  

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Mammalian alkaline phosphatase (ALP) is attached to the plasma membrane by a unique glycosylphosphatidylinositol (GPI) anchor. The influence of such a complex anchoring device on the enzyme function is not fully understood. Here, we report the effect of cleavage of the GPI anchor on the activity of goat liver plasma membrane ALP (GLPM-ALP). Phosphatidylinositol-specific phospholipase C (PI-PLC) purified from Bacillus cereus was used for the cleavage of the GPI anchor (delipidation) and hence for release of ALP from the membrane. Detergents--octyl-beta-D-glucopyranoside (OG) and triton X100 (TX100) were also used for solubilization of ALP from the membrane. Resistance to solubilization by TX100 suggested the association of GPI-ALP with lipid rafts. Solubilization of GLPM-ALP with OG had no effect on the enzyme activity; however, delipidation with PI-PLC resulted in enhanced ALP activity. Kinetic analysis showed catalytic activation of PI-PLC-treated GLPM-ALP with an increase in V(max) (35%) without a significant change in K(m). Moreover, this change in Vmax was observed to be independent of pH and buffer. The results suggested the implication of GPI anchor in modulating the catalytic property of GLPM-ALP, thus indicating the role of this special anchoring structure in the enzyme regulation. PMID:25296497

Kothekar, Deepali; Bandivdekar, Atmaram; Dasgupta, Debjani

2014-08-01

220

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

Science.gov (United States)

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

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

2003-01-01

221

Matrix gla protein and alkaline phosphatase are differently modulated in human dermal fibroblasts from PXE patients and controls.  

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Mineralization of elastic fibers in pseudoxanthoma elasticum (PXE) has been associated with low levels of carboxylated matrix gla protein (MGP), most likely as a consequence of reduced vitamin K (vit K) availability. Unexpectedly, vit K supplementation does not exert beneficial effects on soft connective tissue mineralization in the PXE animal model. To understand the effects of vit K supplementation and in the attempt to interfere with pathways leading to the accumulation of calcium and phosphate within PXE-mineralized soft connective tissues, we have conducted in vitro studies on dermal fibroblasts isolated from control subjects and from PXE patients. Cells were cultured in standard conditions and in calcifying medium (CM) in the presence of vit K1 and K2, or levamisole, an alkaline phosphatase (ALP) inhibitor. Control and PXE fibroblasts were characterized by a similar dose-dependent uptake of both vit K1 and vit K2, thus promoting a significant increase of total protein carboxylation in all cell lines. Nevertheless, MGP carboxylation remained much less in PXE fibroblasts. Interestingly, PXE fibroblasts exhibited a significantly higher ALP activity. Consistently, the mineralization process induced in vitro by a long-term culture in CM appeared unaffected by vit K, whereas it was abolished by levamisole. PMID:23223140

Boraldi, Federica; Annovi, Giulia; Vermeer, Cees; Schurgers, Leon J; Trenti, Tommaso; Tiozzo, Roberta; Guerra, Deanna; Quaglino, Daniela

2013-04-01

222

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

Science.gov (United States)

A commercially available synthetic nucleic acid probe (SNAP) conjugated to alkaline phosphatase was compared with standard culture techniques for detecting Campylobacter species. The SNAP was able to detect either 5 ng of C. jejuni DNA or 10(5) CFU of bacteria. The SNAP could also detect DNA extracted from 10(5) CFU in mock-infected stool samples. The SNAP detected C. jejuni and C. coli but showed no reactivity with C. laridis, C. fetus subsp. fetus, C. fetus subsp. venerealis, C. fennelliae, "C. upsaliensis," C. cinaedii, C. fecalis, C. hyointestinalis, C. mucosalis, or Helicobacter (Campylobacter) pylori. The SNAP also showed no cross-reactivity with other enteric pathogens. When applied to pure cultures, the SNAP detected 55 clinical isolates of C. jejuni and 11 clinical isolates of C. coli, with an accuracy of 100%. When applied directly to clinical specimens, the SNAP detected Campylobacter spp. in 19 of 23 culture-positive stool specimens (sensitivity, 82.6%; specificity, 100%). Pure cultures of the Campylobacter strains isolated from the four probe-negative, culture-positive stool specimens gave positive reactions with the SNAP. While the SNAP had excellent sensitivity and specificity for isolated bacterial colony isolates, the main limitation to the Campylobacter probe detection kit may be the sensitivity limit on direct detection of Campylobacter organisms in stools. PMID:2380381

Olive, D M; Johny, M; Sethi, S K

1990-07-01

223

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

Science.gov (United States)

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

2009-01-01

224

A phage-displayed single domain antibody fused to alkaline phosphatase for detection of porcine circovirus type 2.  

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The fusion of single domain antibodies (sdAbs) and alkaline phosphatase (AP) has been demonstrated to be useful as an immunodiagnostic reagent. In this work, a porcine circovirus type-2 (PCV2) specific sdAb (psdAb) was expressed as fusion with an AP. The binding activity of psdAb-AP fusion was examined by Western blot, enzyme linked immunosorbent assay (ELISA), and surface plasmon resonance (SPR). To assess the practicality of psdAb-AP fusion as a diagnostic reagent, the fusion was used in a Western blot, a direct ELISA, and an immunocytochemistry assay (ICC) for PCV2 detection. The results indicated that the binding activity and specificity of psdAb-AP fusion was similar with psdAb, but the functional affinity of psdAb-AP fusion was about 5 times greater than psdAb as determined by SPR. As a tracer, psdAb-AP fusion could detect PCV2 cap protein down to 0.01?g/lane and 0.05?g/ml in Western blot and direct ELISA respectively. When compared with a control indirect fluorescence assay (IFA), the ICC psdAb-AP fusion was more efficient, needed less operation steps and ended in a shorter time. The results demonstrate that the fusion of psdAb to AP provides a valuable route to the development of psdAb-based immuno-reagents, which offers a simple, convenient, and sensitive method for PCV2 detection. PMID:25512132

Yang, Shunli; Shang, Youjun; Yin, Shuanghui; Wang, Di; Cai, Jianping; Gong, Zhenli; Serge, Muyldermans; Liu, Xiangtao

2015-03-01

225

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

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

Ismini Nakouti and Glyn Hobbs

2012-02-01

226

Zeptomole-detecting biosensor for alkaline phosphatase in an electrochemical immunoassay for 2,4-dichlorophenoxyacetic acid.  

Science.gov (United States)

A bienzyme substrate-recycling biosensor in a flow injection analysis system is described for the sensitive measurement of alkaline phosphatase (ALP) and applied to the fast readout of a competitive immunoassay for the widely used pesticide 2,4-dichlorophenoxyacetic acid (2,4-D). The phenol-indicating biosensor consists of a Clark-type electrode covered by a membrane with coentrapped tyrosinase and quinoprotein glucose dehydrogenase. ALP dephosphorylates phenyl phosphate to phenol (K(m) = 36 microM) outside the flow system. Phenol is oxidized in the sensor membrane by the oxygen-consuming tyrosinase via catechol to o-quinone. The quinone is reconverted to catechol by glucose dehydrogenase. This substrate cycling results in a 350-fold amplified sensor response to phenol. The oxygen consumption of the enzyme couple in the presence of phenol is monitored as a decrease in current. A total of 3.2 fM ALP (320 zmol/ 100 microL) has been detected after a 57.5 min incubation with phenyl phosphate. All involved reagents are stable over the time of measurement. The sensor does not produce any measurable blank signals. The immunoassay detects 0.1 microgram/L 2,4-D, the maximum concentration for pesticides allowed in drinking water by European Community regulations. The applicability of this biosensor for fast immunoassay readout is demonstrated by a 2 min incubation. By comparison, a standard photometric method (p-nitrophenyl phosphate) requires overnight incubation. PMID:8694255

Bauer, C G; Eremenko, A V; Ehrentreich-Förster, E; Bier, F F; Makower, A; Halsall, H B; Heineman, W R; Scheller, F W

1996-08-01

227

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

Energy Technology Data Exchange (ETDEWEB)

Highlights: • Cd{sup 2+} reacts with S{sup 2?} to generate fluorescent CdS QDs with ATP. • ATP can be hydrolyzed by the enzymatic dephosphorylation of ALP. • Fluorescent CdS QDs could not be generated in the presence of ALP. • The analysis system was successfully applied to assay ATP and ALP. - Abstract: We have developed an analytical method to detect adenosine-5?-triphosphate (ATP) and alkaline phosphatase (ALP) based on the generation of CdS quantum dots (QDs). We demonstrated that Cd{sup 2+} cation reacts with S{sup 2?} anion to generate fluorescent CdS QDs in the presence of some certain amount of ATP. With increase in the ATP concentration, the fluorescence intensity of CdS QDs was also enhanced. ATP can be converted into adenosine by the dephosphorylation of ALP, so that the generation of CdS QDs would be inhibited in the presence of ALP. Therefore, this novel analysis system could be applied to assay ATP and ALP based on the growth of fluorescent CdS QDs.

Liu, Siyu; Wang, Xinyan; Pang, Shu; Na, Weidan; Yan, Xu; Su, Xingguang, E-mail: suxg@jlu.edu.cn

2014-05-01

228

Mononuclear and dinuclear peroxotungsten complexes with co-ordinated dipeptides as potent inhibitors of alkaline phosphatase activity.  

Science.gov (United States)

New molecular peroxotungstate(VI) complexes with dipeptides as ancillary ligands of the type, [WO(O(2))(2)(dipeptide)(H(2)O)].3H(2)O, dipeptide = glycyl-glycine or glycyl-leucine, have been synthesized and characterized by elemental analysis, spectral and physico-chemical methods including thermal analysis. The complexes contain side-on bound peroxo groups and a peptide zwitterion bonded to the metal centre unidentately through an O(carboxylate) atom. Investigations on certain biologically important key properties of these compounds and a set of dimeric compounds in analogous co-ligand environment, Na(2)[W(2)O(3)(O(2))(4)(dipeptide)(2)].3H(2)O, dipeptide = glycyl-glycine and glycyl-leucine, reported previously by us revealed interesting features of the compounds. Each of the compounds despite having a 7 co-ordinated metal centre exerts a strong inhibitory effect on alkaline phosphatase activity with a potency higher than that of the free dipeptide, tungstate or peroxotungstate. The compounds exhibit remarkable stability in solutions of acidic as well as physiological pH and are weaker as substrate to the enzyme catalase, compared to H(2)O(2). The mononuclear and dinuclear peroxotungsten compounds are efficient oxidants of reduced glutathione (GSH), a reaction in which only one of the peroxo groups of a diperoxotungsten moiety of the complexes was found to be active. PMID:18665997

Hazarika, Pankaj; Kalita, Diganta; Islam, Nashreen S

2008-08-01

229

Assessment of lactate dehydrogenase, alkaline phosphatase and aspartate aminotransferase activities in cow's milk as an indicator of subclinical mastitis.  

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This study examined the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and aspartate aminotransferase (AST) in the milk of lactating Holstein cows in association with subclinical mastitis (SCM). A total of 94 milk samples were collected from 58 lactating dairy cows representing stages of lactation from the second to the tenth week after calving. Those which were classified as positive by California mastitis test (CMT) were deemed to have subclinical mastitis. All the milk samples were skimmed by centrifugation at 10 000g at 0 degrees C and were used for enzyme activities estimations. The mean activities of LDH and ALP were higher in the milk from udders with SCM than in the milk from healthy udders (p 180 IU/L and > 40 IU/L respectively (kappa values 0.65 and 0.79, respectively). However, the sensitivity of the tests for identifying SCM at these thresholds was higher for ALP (96.4%) than for LDH (68.5%). In this study, LDH and ALP tests were standardized for cow's milk and results showed that only the ALP test was reliable in the early diagnosis of subclinical mastitis. PMID:17268916

Babaei, H; Mansouri-Najand, L; Molaei, M M; Kheradmand, A; Sharifan, M

2007-05-01

230

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

Science.gov (United States)

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

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

2014-05-01

231

Remyelination of spinal cord axons by olfactory ensheathing cells and Schwann cells derived from a transgenic rat expressing alkaline phosphatase marker gene  

OpenAIRE

Transplantation of cell suspensions containing olfactory ensheathing cells (OECs) has been reported to remyelinate demyelinated axons in the spinal cord with a Schwann cell (SC)-like pattern of myelination. However, questions have been raised recently as to whether OECs can form SC-like myelin. To address this issue we prepared SCs and OECs from transgenic rats in which a marker gene, human placental alkaline phosphatase (hPAP), is linked to the ubiquitously active promoter of the R26 gene. S...

Akiyama, Yukinori; Lankford, Karen; Radtke, Christine; Greer, Charles A.; Kocsis, Jeffery D.

2004-01-01

232

Detection of mRNA by in situ hybridisation and in northern blot analysis using oligodeoxynucleotide probes labelled with alkaline phosphatase.  

OpenAIRE

AIMS: To assess whether a reduction in intensity of signal observed using an alkaline phosphatase labelled oligodeoxynucleotide probe could be explained on the basis of procedural steps rather than reduced sensitivity. METHOD: Signal intensity was assessed on in situ hybridisation for pro-opiomelanocortin (POMC) mRNA in rat pituitary and for somatostatin mRNA in human pancreas and in northern blot analysis for POMC mRNA in the presence and absence of formamide. The direct effects of formamide...

Farquharson, M. A.; Harvie, R.; Kennedy, A.; Mcnicol, A. M.

1992-01-01

233

Bacillus subtilis NhaC, an Na+/H+ Antiporter, Influences Expression of the phoPR Operon and Production of Alkaline Phosphatases  

OpenAIRE

When Bacillus subtilis is subjected to phosphate starvation, genes of the Pho regulon are either induced or repressed. Among those induced are genes encoding alkaline phosphatases (APases). A set of isogenic mutants, with a ?-galactosidase gene transcriptionally fused to the inactivated target gene, was used to identify genes that influence the operation of the Pho regulon. One such gene was nhaC (previously yheL). In the absence of NhaC, growth and APase production were enhanced, while the ...

Pra?gai, Zolta?n; Eschevins, Caroline; Bron, Sierd; Harwood, Colin R.

2001-01-01

234

Pyridoxamine-5-phosphate enzyme-linked immune mass spectrometric assay substrate for linear absolute quantification of alkaline phosphatase to the yoctomole range applied to prostate specific antigen.  

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There is a need to measure proteins that are present in concentrations below the detection limits of existing colorimetric approaches with enzyme-linked immunoabsorbent assays (ELISA). The powerful enzyme alkaline phosphatase conjugated to the highly specific bacterial protein streptavidin binds to biotinylated macromolecules like proteins, antibodies, or other ligands and receptors with a high affinity. The binding of the biotinylated detection antibody, with resulting amplification of the signal by the catalytic production of reporter molecules, is key to the sensitivity of ELISA. The specificity and amplification of the signal by the enzyme alkaline phosphatase in ELISA together with the sensitivity of liquid chromatography electrospray ionization and mass spectrometry (LC-ESI-MS) to detect femtomole to picomole amounts of reporter molecules results in an ultrasensitive enzyme-linked immune mass spectrometric assay (ELIMSA). The novel ELIMSA substrate pyridoxamine-5-phosphate (PA5P) is cleaved by the enzyme alkaline phosphatase to yield the basic and hydrophilic product pyridoxamine (PA) that elutes rapidly with symmetrical peaks and a flat baseline. Pyridoxamine (PA) and (13)C PA were both observed to show a linear relationship between log ion intensity and quantity from picomole to femtomole amounts by liquid chromatography-electrospray ionization and mass spectrometry. Four independent methods, (i) internal (13)C isotope PA dilution curves, (ii) internal (13)C isotope one-point calibration, (iii) external PA standard curve, and (iv) external (13)C PA standard curve, all agreed within 1 digit in the same order of magnitude on the linear quantification of PA. Hence, a mass spectrometer can be used to robustly detect 526 ymol of the alkaline phosphatase streptavidin probe and accurately quantify zeptomole amounts of PSA against log linear absolute standard by micro electrospray on a simple ion trap. PMID:25259405

Florentinus-Mefailoski, Angelique; Marshall, John G

2014-11-01

235

Human intestinal alkaline phosphatase--release to the blood is linked to lipid absorption, but removal from the blood is not linked to lipoprotein clearance.  

OpenAIRE

To evaluate a possible quantitative relationship between the rise in intestinal alkaline phosphatase (IAP) activity and triglyceride-rich lipoproteins in the blood after an oral fat intake, a specific and sensitive immunocatalytic assay was used. First, day to day variation of the basal IAP activity in the blood of eight volunteers was evaluated. One group of subjects with high basal IAP activity and great variations from one day to the other was distinguished from a group with low basal IAP ...

Domar, U.; Karpe, F.; Hamsten, A.; Stigbrand, T.; Olivecrona, T.

1993-01-01

236

Studies on alkaline and acid phosphatase activity of neutrophil leukocytes, 2. Influence of anticancer drugs or /sup 60/Co irradiation on alkaline and acid phosphatase activity of neutrophils (pseudoeosinophil) in rabbits  

Energy Technology Data Exchange (ETDEWEB)

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 enzyme 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 hematological 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) /sup 60/Co 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.

Niki, Yoko (Kansai Medical School, Moriguchi, Osaka (Japan))

1983-03-01

237

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

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

T. Raeisi

2014-07-01

238

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

International Nuclear Information System (INIS)

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

239

Pst I restriction fragment length polymorphism of human placental alkaline phosphatase gene: Mendelian in segregation and localization of mutation site in the gene  

International Nuclear Information System (INIS)

The pattern of inheritance of a Pst I restriction fragment length polymorphism (RFLP) of the human placental alkaline phosphatase gene was studied in nine nuclear families by Southern blot hybridization analysis of genomic DNA. The dimorphic RFLP is defined by the presence of allelic fragments 1.0 kilobase and 0.8 kilobase long. The results of this study show that the two alleles of the Pst I RFLP of the placental alkaline phosphatase gene segregate as codominant traits according to Mendelian expectations. For a polymorphism to be useful as a genetic marker the probability that an offspring is informative (PIC) must be at least 0.15. The allelic frequency of the 1.0-kilobase allele is 0.21, which correlates to a probability that an offspring is informative of 0.275 and is indicative of a useful polymorphism. By using probes derived from different regions of the placental alkaline phosphatase cDNA, the mutated Pst I site causing the RFLP was located in the penultimate intron 2497 base pairs downstream from the transcriptional initiation site

240

Age-related changes of plasma alkaline phosphatase and inorganic phosphorus, and late ossification of the cranial roof in the spanish imperial eagle (Aquila adalberti C. L. Brehm, 1861)  

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Plasma alkaline phosphatase and inorganic phosphorus levels were determined for 52 nestling Spanish imperial eagles from two wild populations and 22 captive adults and subadults (10 adults and 12 subadults). The exact age was known for all birds. Mean alkaline phosphatase and inorganic phosphorus were higher in chicks than in the captive adults and subadults. Sex differences were not observed, and nestlings from different populations showed similar values. No significant regression described ...

Dobado-berrios, P. M.; Ferrer, Miguel

1997-01-01

241

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  

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Full Text Available 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 (inmunoturbidimétrico, proteiunuria (turbidimétrico, uroproteinograma, SDS-PAGE (al 12,5% e isoenzimograma de FAL. La FAL sérica hallada fue normal, sin diferencia entre grupos y sin relación con el aumento de la FALur. El valor de corte recomendado para FALur fue de 8 UI/L. La FALur estuvo elevada en HTA e IRDO y normal en individuos con DBT. Los aumentos de FALur en IRDO se relacionaron con la lesión tubular estructural y en pacientes con HTA podrían relacionarse con alteración tubular precoz. Se propone la determinación de FALur para la detección temprana de lesión tubular ante falla renal establecida o en individuos con riesgo de desarrollarla, y se establece su utilidad en pacientes: - con DBT y HTA para seguimiento (junto a microalbuminuria y clearence de creatinina, - internados en riesgo de insuficiencia renal aguda: para orientar tratamientos, - con insuficiencia renal crónica: como indicador de lesión y pronóstico.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, uroproteinogram, SDS-PAGE (12.5% and ALP isoenzymes determinations were made. The results indicate that serum ALP was normal, without difference between groups, and no relation with the increase in ALPur. Recommended cut-off value of ALPur was 8 UI/L. ALPur was elevated in HTA and IRDO, and normal in DBT. Increases in ALPur in IRDO were related to the structural tubular injury, and those in HTA could be related to early tubular alteration. Determination of ALPur is proposed for early detection of tubular injury, before renal failure is established or when there is risk of developing it, establishing its usefulness in: - DBT and HTA patients: screening (together with microalbuminuria and creatinine clearence. - Hospitalized patients in risk of acute renal insufficiency: in order to orient treatments. - Patients with chronic renal insufficiency: as an indicator of injury and prognosis.

María Beatriz Di Carlo

2007-09-01

242

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

2005-08-01

243

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

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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 18h 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.

2008-01-01

244

A comparative study of two different assay kits for the detection of secreted alkaline phosphatase in HPV antibody neutralization assays.  

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To assess immunogenicity and development of antibodies in the context of vaccination, it is critical to quantify titers of neutralizing antibodies. We have been employing the 293TT cell-based neutralization assay system to quantify anti-HPV neutralizing antibodies. In this system, human papillomavirus (HPV) pseudovirion (PsV) particles encapsidating secreted alkaline phosphatase (SEAP) gene are used to measure infection of 293TT cells in 72-hr cell-culture supernatants. SEAP has traditionally been measured by Great EscAPe™ SEAP Chemiluminescence Kit 2.0 (GE). To reduce the cost, and to potentially increase efficiency, we sought a cheaper kit with better detection capability. Performance characteristics of the newer chemiluminescence kit, ZiVa® Ultra SEAP Plus Assay (Ziva) and GE were compared using the 293TT system. Dose titration of HPV PsV 16 or 18 showed that signal-to-noise ratios at 48 and 72 hr post-infection were higher for ZiVa at nearly all doses. ZiVa was superior to GE as it was able to detect SEAP at 48 hr, as well as when lower numbers of 293TT cells were used. The ability of ZiVa to quantitate HPV-16 and -18 neutralizing antibody titers was tested using sera from Cervarix® immunized individuals. Spearman rank correlational analyses showed excellent correlations between the titers obtained with ZiVa and GE for anti-HPV16 (r = 0.9822, p anti-HPV18 (r = 0.9832, p antibodies. We concluded that ZiVa is superior to GE in detecting SEAP, and the antibody titers in sera of vaccinated individuals were similar to those obtained with GE. Thus, Ziva is a suitable alternative to GE. PMID:25695397

Kemp, Troy J; Matsui, Ken; Shelton, Gloriana; Safaeian, Mahboobeh; Pinto, Ligia A

2015-02-01

245

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

International Nuclear Information System (INIS)

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

246

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

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

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

2013-12-01

247

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

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

MasahiroSuzumura

2012-03-01

248

High-resolution analysis of Zn(2+) coordination in the alkaline phosphatase superfamily by EXAFS and x-ray crystallography.  

Science.gov (United States)

Comparisons among evolutionarily related enzymes offer opportunities to reveal how structural differences produce different catalytic activities. Two structurally related enzymes, Escherichia coli alkaline phosphatase (AP) and Xanthomonas axonopodis nucleotide pyrophosphatase/phosphodiesterase (NPP), have nearly identical binuclear Zn(2+) catalytic centers but show tremendous differential specificity for hydrolysis of phosphate monoesters or phosphate diesters. To determine if there are differences in Zn(2+) coordination in the two enzymes that might contribute to catalytic specificity, we analyzed both x-ray absorption spectroscopic and x-ray crystallographic data. We report a 1.29-Å crystal structure of AP with bound phosphate, allowing evaluation of interactions at the AP metal site with high resolution. To make systematic comparisons between AP and NPP, we measured zinc extended x-ray absorption fine structure for AP and NPP in the free-enzyme forms, with AMP and inorganic phosphate ground-state analogs and with vanadate transition-state analogs. These studies yielded average zinc-ligand distances in AP and NPP free-enzyme forms and ground-state analog forms that were identical within error, suggesting little difference in metal ion coordination among these forms. Upon binding of vanadate to both enzymes, small increases in average metal-ligand distances were observed, consistent with an increased coordination number. Slightly longer increases were observed in NPP relative to AP, which could arise from subtle rearrangements of the active site or differences in the geometry of the bound vanadyl species. Overall, the results suggest that the binuclear Zn(2+) catalytic site remains very similar between AP and NPP during the course of a reaction cycle. PMID:22056344

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

2012-01-01

249

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

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

Gabriella Caruso

2010-03-01

250

ALP (Alkaline Phosphatase) Test  

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... may be diagnosed and/or monitored using other tests such as calcium , phosphorus , parathyroid hormone , vitamin D , or bone markers (a group of tests used to measure bone formation and bone resorption ). ^ ...

251

The expression of placental alkaline phosphatase (PLAP) and PLAP-like enzymes in normal and neoplastic human tissues. An immunohistological survey using monoclonal antibodies  

DEFF Research Database (Denmark)

The immunohistological expression of placental alkaline phosphatase (PLAP) and PLAP-like enzyme was studied in frozen sections from a wide variety (n = 254) of normal and malignant tissues using monoclonal antibodies reactive with PLAP (H317) and PLAP/PLAP-like enzyme (H17E2; H315). PLAP/PLAP-like reactivity was seen in normal thymus, and foetal and neonatal testis, and in 21 out of 22 malignant germ cell tumours (GCTs), but was also found in normal endocervix, normal Fallopian tube and in 28 out of 167 non-GCTs (particularly in ovarian and proximal gastrointestinal tract tumours). Positivity for true PLAP (as demonstrated with H317) was seen in term placenta, in endocervix, and in Fallopian tube (but not in other normal tissues) and was commonly found in ovarian and proximal gastrointestinal tract tumours. Reactivity with H317 was unusual in malignant GCTs (2 out of 22 cases). These findings confirm that PLAP/PLAP-like positivity is a highly sensitive immunohistological marker for malignant GCTs, but one which by itself is of only moderate specificity. Furthermore, expression of true PLAP is rare in GCTs and favours instead an origin from the ovary or proximal gastrointestinal tract. The results also indicate that the predominant heat-stable alkaline phosphatase species in normal foetal and neonatal testis, and in thymus has a similar immunohistological profile to that found in malignant GCTs, and is a PLAP-like enzyme ("germ cell alkaline phosphatase") distinct from true PLAP. The occurrence of this marker in GCTs would appear to reflect increased eutopic production of an enzyme present in trace amount in corresponding normal tissues rather than a genuine example of ectopic expression.

Hamilton-Dutoit, Stephen Jacques; Lou, H

1990-01-01

252

Resveratrol Increases Bone Mineral Density and Bone Alkaline Phosphatase in Obese Men : A Randomized Placebo-Controlled Trial  

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Context: Metabolic syndrome (MetS) is associated with low-grade inflammation, which may harmfully affect bone. Resveratrol (RSV) possesses anti-inflammatory properties, and rodent studies suggest bone protective effects. Objective: This study sought to evaluate effects of RSV treatment on bone in men with MetS. Setting and Design: The study was conducted at Aarhus University Hospital as a randomized, double-blinded, placebo-controlled trial assessing changes in bone turnover markers, bone mineral density (BMD), and geometry. Participants: The study population comprised 74 middle-aged obese men with MetS recruited from the general community, of which 66 completed all visits. Mean age of participants was 49.3 ± 6.3 years and mean body mass index was 33.7 ± 3.6 kg/m(2). Intervention: Oral treatment with 1.000 mg RSV (RSVhigh), 150mg RSV (RSVlow), or placebo daily for 16 weeks. Main Outcome Measure: Prespecified primary endpoint was change in bone alkaline phosphatase (BAP). Results: BAP increased dose dependently with RSV (R = 0.471, P < .001), resulting in a significantly greater increase in BAP in the RSVhigh group compared with placebo at all time-points (week 4, 16.4 ± 4.2%, P < .001; week 8, 16.5 ± 4.1%, P < .001; week 16, 15.2 ± 3.7%, P < .001). Lumbar spine trabecular volumetric bone mineral density (LS vBMDtrab) also increased dose dependently with RSV (R = 0.268, P = .036), with a significant increase of 2.6 ± 1.3% in the RSVhigh group compared with placebo (P = .043). In addition, changes in BAP and LS vBMDtrab were positively correlated (R = 0.281, P = .027). No consistent changes were detected in bone density at the hip. Conclusions: Our data suggest that high-dose RSV supplementation positively affects bone, primarily by stimulating formation or mineralization. Future studies of longer duration comprising populations at risk of osteoporosis are needed to confirm these results.

Ornstrup, Marie Juul; HarslØf, Torben

2014-01-01

253

In vivo overexpression of tissue-nonspecific alkaline phosphatase increases skeletal mineralization and affects the phosphorylation status of osteopontin.  

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Functional ablation of tissue-nonspecific alkaline phosphatase (TNAP) (Alpl?/? mice) leads to hypophosphatasia, characterized by rickets/osteomalacia attributable to elevated levels of extracellular inorganic pyrophosphate, a potent mineralization inhibitor. Osteopontin (OPN) is also elevated in the plasma and skeleton of Alpl?/? mice. Phosphorylated OPN is known to inhibit mineralization, however, the phosphorylation status of the increased OPN found in Alpl?/? mice is unknown. Here, we generated a transgenic mouse line expressing human TNAP under control of an osteoblast-specific Col1a1 promoter (Col1a1-Tnap). The transgene is expressed in osteoblasts, periosteum, and cortical bones, and plasma levels of TNAP in mice expressing Col1a1-Tnap are 10 to 20 times higher than those of wild-type mice. The Col1a1-Tnap animals are healthy and exhibit increased bone mineralization by micro-computed tomography (µCT) analysis. Crossbreeding of Col1a1-Tnap transgenic mice to Alpl?/? mice rescues the lethal hypophosphatasia phenotype characteristic of this disease model. Osteoblasts from [Col1a1-Tnap] mice mineralize better than nontransgenic controls and osteoblasts from [Col1a1-Tnap?/?; Alpl?/?] mice are able to mineralize to the level of Alpl?/? heterozygous osteoblasts, whereas Alpl?/? osteoblasts show no mineralization. We found that the increased levels of OPN in bone tissue of Alpl?/? mice are comprised of phosphorylated forms of OPN whereas wild-type (WT) and [Col1a1-Tnap?/?; Alpl?/?] mice had both phosphorylated and dephosphorylated forms of OPN. OPN from [Col1a1-Tnap] osteoblasts were more dephosphorylated than nontransgenic control cells. Titanium dioxide-liquid chromatography and tandem mass spectrometry analysis revealed that OPN peptides derived from Alpl?/? bone and osteoblasts yielded a higher proportion of phosphorylated peptides than samples from WT mice, and at least two phosphopeptides, p(S¹??FQVS¹??DEQY¹?²PDAT¹??DEDLT¹?¹)SHMK and FRIp(S²??HELES³??S³??S³??S³??)EVN, with one nonlocalized site each, appear to be preferred sites of TNAP action on OPN. Our data suggest that the promineralization role of TNAP may be related not only to its accepted pyrophosphatase activity but also to its ability to modify the phosphorylation status of OPN. PMID:23427088

Narisawa, Sonoko; Yadav, Manisha C; Millán, José Luis

2013-07-01

254

Cycling of Dissolved Organic Phosphorus and Alkaline Phosphatase Activity in Euphotic Zone of the Western North Pacific  

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Phosphorus is an essential nutrient for marine organisms. In oligotrophic environments, concentrations of dissolved inorganic phosphate (SRP), the most bioavailable form of phosphorus, are low and have been hypothesized to constrain the primary productivity. Evidence has been found that dissolved organic phosphorus (DOP) supports a significant fraction of primary production through hydrolytic remineralization of DOP to SRP by alkaline phosphatase (APA). In this study, DOP biogeochemistry was investigated at three locations of the open-ocean environment in the Kuroshio region and at a semi-eutrophic coastal site of the western North Pacific. Concentrations of SRP, DOP and hydrolyzable ester-P were measured in the euphotic zone. Kinetic parameters of APA were determined using a fluorogenic substrate, including potential maximum velocity (Vmax), apparent Michaelis-Menten half-saturation constant (Km), and turnover time (TA) of APA hydrolyzable DOP. SRP concentrations were quite low (? 10 nM) in the surface seawater and rapidly increased below the chlorophyll a maximum layer (CML). DOP concentration ranged from 29 to 223 nM. Above the CML, DOP composed a major fraction accounting for 60-100% of dissolved total P. A significant linear relationship was found between the concentrations of SRP and hydrolyzable ester-P (R2 = 0.83, P < 0.01). This suggests active utilization of ester-P under phosphate-depleted conditions. In the Kuroshio region, Vmax of APA exhibited the highest value at the surface water (0 m) and decreased rapidly with depth, while at the coastal site the peak value was found at CML. TA of hydrolyzable DOP was quite variable among the locations and increased with depth especially below CML. The estimated values of in situ hydrolysis rate were much lower (2-34%) than the potential Vmax which was determined with the addition of an excess amount of the substrate. The results suggest that marine microbes can efficiently and rapidly utilize hydrolyzable DOP under phosphate-depleted conditions and that there is still room in the in situ APA activity. Utilization of DOP, however, is likely regulated by the ambient concentrations of hydrolyzable ester-P lower than the apparent Km.

Suzumura, M.

2010-12-01

255

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

Scientific Electronic Library Online (English)

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

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

2008-01-01

256

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

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Full Text Available Background and Objective: The enamel matrix derivative (EMD has been used in combination with barrier membranes to optimize regeneration in vertical osseous defects. However, the osteoblast response when exposed to the EMD/barrier membrane combination has not yet been evaluated. The osteoblast behavior when exposed to a combination of regenerative materials must be evaluated to fully understand their effect on bone regeneration. Therefore, the present study was undertaken to estimate the initial adhesion and alkaline phosphatase (ALP activity of an osteoblast cell line (SaOS-2 when exposed to four commercially available resorbable membranes and determine if the addition of EMD had any modulatory effect on osteoblast behavior. Materials and Methods: 5 x 104 SaOS-2 cells between passages 7-10 were cultured in two 24-well culture plates. Plate A was used for the adhesion assay and Plate B was used for the ALP assay. A MTT (3-[4, 5-dimethylthiazolyl-2]-2, 5-diphenyltetrazolium bromide assay was done after 24 hours to determine the adhesion of the osteoblastic cells to four barrier membranes: 1 a non cross-linked porcine Type I and III collagen membrane (BG, 2 a weakly cross-linked Type I collagen membrane (HG, 3 a glutaraldehyde cross-linked bovine Type I collagen (BM, and 4 a resorbable polymer membrane (CP. Osteoblast differentiation was studied using an ALP assay with p-nitro phenyl phosphate as the substrate at 24 hours, 72 hours, and 1 week. A total of 50 µg/ml of EMD dissolved in 10 mM acetic acid was added into each well and the entire experimental protocol outlined above was repeated. Results: The osteoblast adhesion to collagen barriers showed a statistically insignificant reduction following the addition of EMD. Adhesion to the polymer barrier, although significantly lower when compared with collagen barriers, was unaffected by the addition of EMD. ALP activity after 1 week among the various groups was as follows: EMD alone (75.59±2.5>EMD/BG(64.78±3.04>EMD/HG(55.40±3.89?EMD/BM(54.75±4.17>BG (51.32±2.76>HG(49.92±2.4>BM(48.14±1.4>Control(46.29±1.39>EMD/CP (37.46±3.54>CP(32.12±1.49 Conclusion: There was no additive effect on osteoblast adhesion/ALP activity following exposure to an EMD/polymer combination. EMD/collagen positively influences osteoblast differentiation in a time dependent manner.

Thangakumaran S

2009-01-01

257

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)

258

Effects of dietary vitamin E on mucosal maltase and alkaline phosphatase enzyme activities and on the amount of mucosal malonyldialdehyde in broiler chickens  

OpenAIRE

The effects of dietary vitamin E levels on mucosal maltase and alkaline phosphatase (ALP) enzyme activities and on the amount of mucosal malonyldialdehyde (MDA) in broiler chickens were studied in the present study. One hundred and eighty of day old male broiler chicks (Ross 308 strain) were randomly assigned into five groups, each with three replicates and 12 chicks in each replicate. Chickens in group A were fed corn-soy- based diet, while those in groups B, C, D and E were fed the same die...

Seyed Hamid Farrokhifar; Ramezan Ali Jafari; Naeem Erfani Majd; Seyed Reza Fatemi Tabatabaee; Mansour Mayahi

2013-01-01

259

A study on the pattern of alkaline phosphatase activity correlated with observations on silver-impregnated structures in the developing mouse brain.  

OpenAIRE

Alkaline phosphatase activity was localised in specific groups of cellular processes, nuclei and fibre tracts in fetal mouse brain at 9.5-17.5 days post coitum. A radial band of intense enzymatic activity extended longitudinally within the rhombencephalon at 10.5 to 15.5 days. This enzymatic band was topographically related to the efferent nuclei of the cranial nerves, except those nuclei which had migrated laterally to their final positions. The enzymatic activity was seen in some but not al...

Tam, P. P.; Kwong, W. H.

1987-01-01

260

Remyelination of spinal cord axons by olfactory ensheathing cells and Schwann cells derived from a transgenic rat expressing alkaline phosphatase marker gene.  

Science.gov (United States)

Transplantation of cell suspensions containing olfactory ensheathing cells (OECs) has been reported to remyelinate demyelinated axons in the spinal cord with a Schwann cell (SC)-like pattern of myelination. However, questions have been raised recently as to whether OECs can form SC-like myelin. To address this issue we prepared SCs and OECs from transgenic rats in which a marker gene, human placental alkaline phosphatase (hPAP), is linked to the ubiquitously active promoter of the R26 gene. SCs were prepared from the sciatic nerve and OECs from the outer nerve-fiber layer of the olfactory bulb. Positive S100 and p75 immunostaining indicated that >95% of cells in culture displayed either SC or OEC phenotypes. Suspensions of either SCs or OECs were transplanted into an X-irradiation/ethidium bromide demyelinating lesion in the spinal cord. We observed extensive SC-like remyelination following either SC or OEC transplantation 3 weeks after injection of the cells. Alkaline phosphatase (ALP) chromagen reaction product was associated clearly with the myelin-forming cells. Thus, cell suspensions that are enriched in either SCs or OECs result in peripheral-like myelin when transplanted in vivo. PMID:16799702

Akiyama, Yukinori; Lankford, Karen; Radtke, Christine; Greer, Charles A; Kocsis, Jeffery D

2004-02-01

261

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

Energy Technology Data Exchange (ETDEWEB)

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

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

2010-08-27

262

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

International Nuclear Information System (INIS)

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

263

Serum Dickkopf-1 Level in Postmenopausal Females: Correlation with Bone Mineral Density and Serum Biochemical Markers  

OpenAIRE

Objective. To assess serum level of Dickkopf-1 in postmenopausal females and its correlation with bone mineral density and serum biochemical markers. Methods. Bone densitometry, serum Dickkopf-1, calcium, phosphorus, and alkaline phosphatase were done in sixty postmenopausal females. Patients were divided according to T score into osteoporosis (group I), osteopenia (group II), and normal bone mineral density that served as controls. Results. There was highly significant increase in serum Dick...

Sahar Fathi Ahmed; Neveen Fouda; Amal Ahmed Abbas

2013-01-01

264

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

Directory of Open Access Journals (Sweden)

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

2006-01-01

265

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

2008-04-01

266

Competitive, uncompetitive, and mixed inhibitors of the alkaline phosphatase activity associated with the isolated brush border membrane of the tapeworm Hymenolepis diminuta.  

Science.gov (United States)

Several compounds were tested as inhibitors of the alkaline phosphatase (AlkPase) activity associated with the isolated brush border membrane of the tapeworm, Hymenolepis diminuta. Molybdate, arsenate, arsenite and beta-glycerophosphate (BGP) were competitive inhibitors of the hydrolysis of p-nitrophenyl phosphate, while levamisole and clorsulon were uncompetitive and mixed inhibitors, respectively. Molybdate was also a competitive inhibitor of the hydrolysis of BGP and 5'-adenosine monophosphate, and levamisole was an uncompetitive inhibitor of BGP hydrolysis. The apparent inhibitor constants (Ki') for molybdate and levamisole were virtually identical regardless of the substrate, and these data support the hypothesis that the AlkPase activity is represented by a single membrane-bound enzyme with low substrate specificity. Quinacrine, Hg2+, and ethylenediaminetetraacetate were also potent inhibitors of the AlkPase activity, but the mechanisms by which these latter three inhibitors function were not clear. PMID:2768348

Pappas, P W; Leiby, D A

1989-06-01

267

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

International Nuclear Information System (INIS)

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

268

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

Science.gov (United States)

This paper presents the use of polymer coated microelectrodes for the realtime conductivity monitoring in a microchannel photoablated through the polymer without contact. Based on this strategy, a small conductometry sensor has been developed to record in time conductivity variation when an enzymatic reaction occurs through the channel. The rate constant determination, k2, for the dephosphorylation of organic phosphate-alkaline phosphatase-superparamagnetic beads complex using chemically different substrates such as adenosine monoesterphosphate, adenosine diphosphate and adenosine triphosphate was taken as an example to demonstrate selectivity and sensivity of the detection scheme. The k2 value measured for each adenosine phosphate decreases from 39 to 30 s(-1) in proportion with the number (3, 2 and 1) of attached phosphate moiety, thus emphasizing the steric hindrance effect on kinetics. PMID:25476378

Kechadi, Mohammed; Sotta, Bruno; Gamby, Jean

2015-01-01

269

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

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Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders. PMID:22743140

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

2013-03-01

270

Production and characterization of a single-chain variable fragment linked alkaline phosphatase fusion protein for detection of O,O-diethyl organophosphorus pesticides in a one-step enzyme-linked immunosorbent assay  

Science.gov (United States)

A single-chain variable fragment (scFv) and alkaline phosphatase (AP) fusion protein for detection of O, O-diethyl organophosphorus pesticides (OPs) was produced and characterized. The scFv gene was prepared by cloning VL and VH genes from a hybridoma cell secreting monoclonal antibody with broad-s...

271

Improvement of the skeletal and dental hypophosphatasia phenotype in Alpl-/- mice by administration of soluble (non-targeted) chimeric alkaline phosphatase.  

Science.gov (United States)

Hypophosphatasia (HPP) results from ALPL gene mutations, which lead to a deficiency of tissue-nonspecific alkaline phosphatase (TNAP), and accumulation of inorganic pyrophosphate, a potent inhibitor of mineralization that is also a natural substrate of TNAP, in the extracellular space. HPP causes mineralization disorders including soft bones (rickets or osteomalacia) and defects in teeth and periodontal tissues. Enzyme replacement therapy using mineral-targeting recombinant TNAP has proven effective in preventing skeletal and dental defects in TNAP knockout (Alpl(-/-)) mice, a model for life-threatening HPP. Here, we show that the administration of a soluble, intestinal-like chimeric alkaline phosphatase (ChimAP) improves the manifestations of HPP in Alpl(-/-) mice. Mice received daily subcutaneous injections of ChimAP at doses of 1, 8 or 16 mg/kg, from birth for up to 53 days. Lifespan and body weight of Alpl(-/-) mice were normalized, and vitamin B6-associated seizures were absent with 16 mg/kg/day of ChimAP. Radiographs, ?CT and histological analyses documented improved mineralization in cortical and trabecular bone and secondary ossification centers in long bones of ChimAP16-treated mice. There was no evidence of craniosynostosis in the ChimAP16-treated mice and we did not detect ectopic calcification by radiography and histology in the aortas, stomachs, kidneys or lungs in any of the treatment groups. Molar tooth development and function improved with the highest ChimAP dose, including enamel, dentin, and tooth morphology. Cementum remained deficient and alveolar bone mineralization was reduced compared to controls, though ChimAP-treated Alpl(-/-) mice featured periodontal attachment and retained teeth. This study provides the first evidence for the pharmacological efficacy of ChimAP for use in the treatment of skeletal and dental manifestations of HPP. PMID:25433339

Gasque, Kellen C S; Foster, Brian L; Kuss, Pia; Yadav, Manisha C; Liu, Jin; Kiffer-Moreira, Tina; van Elsas, Andrea; Hatch, Nan; Somerman, Martha J; Millán, José Luis

2015-03-01

272

Sequential preparation of highly purified microvillous and basal syncytiotrophoblast membranes in substantial yield from a single term human placenta: inhibition of microvillous alkaline phosphatase activity by EDTA.  

Science.gov (United States)

The human placental syncytiotrophoblast is a highly polarised epithelial layer responsible for regulating materno-fetal exchange. We here describe a novel procedure for isolating paired fractions of the maternal-facing and fetal-facing plasma membranes from this syncytium, from a single placenta, without the need for homogenisation procedures. This reduces the potential for contamination of these membrane fractions by intracellular membranes, or from plasma membranes from other cell types within the placenta. Microvillous membrane vesicles (MVM) were obtained by gentle stirring of dispersed villous tissue. The tissue sedimented at the end of this procedure was subjected to sequential ultrasonication to release the basal membrane (BM). Crude MVM was subsequently purified on a discontinuous sucrose gradient. Crude BM was further purified using either discontinuous Ficoll or sucrose gradients. The Ficoll procedure, while producing a BM fraction extremely enriched in marker enzyme, resulted in unacceptably low protein recoveries and hence the sucrose gradient procedure was also adopted for BM. Yields for MVM and BM produced on sucrose density gradients approached 30 mg/100 g tissue. The MVM fraction was composed of vesicles of 232 +/- 9 (S.E.) nm diameter of which nearly 90% were 'right side out'. These membranes were 37-fold enriched in the marker enzyme alkaline phosphatase. Purified BM vesicles were 317 +/- 14 nm in diameter, also approximately 90% 'right side out' and over 40-fold enriched in dihydroalprenolol binding. Cross-contamination or contamination from intracellular membranes was negligible. MVM alkaline phosphatase activity was shown to be inhibitable in a dose- and time-dependent manner by EDTA present in the storage buffer. PMID:8038198

Eaton, B M; Oakey, M P

1994-07-13

273

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

Scientific Electronic Library Online (English)

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

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

2011-12-01

274

Relation between serum PAP (prostate acid phosphatase) and bone scintigraphy in prostatic cancer  

International Nuclear Information System (INIS)

Seventy-seven patients with prostatic cancer were treated at our department in the last 5 years. Of these patients 30 cases were followed by bone scintigraphy and serum PAP. In 27 follow-up scintigraphy procedures changes of bone scintigraphy corresponded to changes in serum PAP levels. Changes of PAP levels did not always correspond to changes of scintigraphy, but almost all cases in which the level of PAP increased in a short period showed progression of bone metastasis. A 3-month interval between bone scintigraphy procedure in stage D2 prostatic cancer patients is generally recommended. However, we think that in prostatic cancer patients follow-up bone scintigraphy at regular short intervals is unnecessary if there is no change in serum PAP levels, symptoms or physical condition. Bone scintigraphy should be performed when the tumor marker changes rapidly or when any physical symptom appears. (author)

275

Relation between serum PAP (prostate acid phosphatase) and bone scintigraphy in prostatic cancer  

Energy Technology Data Exchange (ETDEWEB)

Seventy-seven patients with prostatic cancer were treated at our department in the last 5 years. Of these patients 30 cases were followed by bone scintigraphy and serum PAP. In 27 follow-up scintigraphy procedures changes of bone scintigraphy corresponded to changes in serum PAP levels. Changes of PAP levels did not always correspond to changes of scintigraphy, but almost all cases in which the level of PAP increased in a short period showed progression of bone metastasis. A 3-month interval between bone scintigraphy procedure in stage D2 prostatic cancer patients is generally recommended. However, we think that in prostatic cancer patients follow-up bone scintigraphy at regular short intervals is unnecessary if there is no change in serum PAP levels, symptoms or physical condition. Bone scintigraphy should be performed when the tumor marker changes rapidly or when any physical symptom appears. (author).

Aizawa, Taku; Itoh, Takaaki; Tsujino, Susumu; Namiki, Kazunori; Miki, Makoto (Tokyo Medical Coll. (Japan))

1992-11-01

276

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

277

Tissue-nonspecific alkaline phosphatase acts redundantly with PAP and NT5E to generate adenosine in the dorsal spinal cord.  

Science.gov (United States)

Prostatic acid phosphatase (PAP) and ecto-5'-nucleotidase (NT5E) hydrolyze extracellular AMP to adenosine in dorsal root ganglia (DRG) neurons and in the dorsal spinal cord. Previously, we found that adenosine production was reduced, but not eliminated, in Pap?/?/Nt5e?/? double knock-out (dKO) mice, suggesting that a third AMP ectonucleotidase was present in these tissues. Here, we found that tissue-nonspecific alkaline phosphatase (TNAP, encoded by the Alpl gene) is expressed and functional in DRG neurons and spinal neurons. Using a cell-based assay, we found that TNAP rapidly hydrolyzed extracellular AMP and activated adenosine receptors. This activity was eliminated by MLS-0038949, a selective pharmacological inhibitor of TNAP. In addition, MLS-0038949 eliminated AMP hydrolysis in DRG and spinal lamina II of dKO mice. Using fast-scan-cyclic voltammetry, we found that adenosine was rapidly produced from AMP in spinal cord slices from dKO mice, but virtually no adenosine was produced in spinal cord slices from dKO mice treated with MLS-0038949. Last, we found that AMP inhibited excitatory neurotransmission via adenosine A1 receptor activation in spinal cord slices from wild-type, Pap?/?, Nt5e?/?, and dKO mice, but failed to inhibit neurotransmission in slices from dKO mice treated with MLS-0038949. These data suggest that triple elimination of TNAP, PAP, and NT5E is required to block AMP hydrolysis to adenosine in DRG neurons and dorsal spinal cord. Moreover, our data reveal that TNAP, PAP, and NT5E are the main AMP ectonucleotidases in primary somatosensory neurons and regulate physiology by metabolizing extracellular purine nucleotides. PMID:23825434

Street, Sarah E; Kramer, Nicholas J; Walsh, Paul L; Taylor-Blake, Bonnie; Yadav, Manisha C; King, Ian F; Vihko, Pirkko; Wightman, R Mark; Millán, José Luis; Zylka, Mark J

2013-07-01

278

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

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

Kala Mrinalini

2005-12-01

279

Correlation between the extent of metastatic lesions in whole body bone scintigraphy of patients with prostatic cancer and prostatic acid phosphatase levels in serum with Eiken PAP RIA kit  

International Nuclear Information System (INIS)

The whole body bone scintigraphy of thirteen patients whose prostatic cancer were histologically confirmed, was processed in four colors, and the bone metastases were quantitatively estimated. On the basis of this estimation, the extent of bone metastases was classified into 4 divisions (grades 0, 1, 2 and 3). And then, the correlation between the extent of bone metastases and prostatic acid phosphatase (PAP), acid phosphatase (AcP) and alkali phosphatase (AlP) levels in serum were investigated

280

Polyphenolic compounds from flowers of Hibiscus rosa-sinensis Linn. and their inhibitory effect on alkaline phosphatase enzyme activity in vitro.  

Science.gov (United States)

Graded concentrations (0.1-100 mg/mL reaction mixture) of the methanolic extract of the flowers of Hibiscus rosa-sinensis Linn., its water-soluble fraction as well as compounds isolated from this fraction were tested for their inhibitory effect on alkaline phosphatase enzyme activity in vitro. Both the methanolic extract and its water-soluble fraction showed significant inhibitory effects on the enzyme activity in vitro. On screening the activity of the compounds isolated from the water-soluble fraction, its high inhibitory activity was attributed to the presence of quercetin-7-O-galactoside which showed a high potent inhibition of the enzyme activity reaching 100% at 100 mg/mL reaction mixture. Phytochemical investigations of the water-soluble fraction were also carried out and afforded ten polyphenolic compounds including two new natural compounds, namely kaempferol-7-O-[6'''-O-p-hydroxybenzoyl-beta-D-glucosyl-(1-->6)-beta-D-glucopyranoside] and scutellarein-6-O-alpha-L-rhamnopyranoside-8-C-beta-D-glucopyranoside). The chemical structure of the isolated compounds was elucidated on the basis of chemical and spectral data. PMID:22191209

Salib, Josline Y; Daniel, Enas N; Hifnawy, Mohamed S; Azzam, Shadia M; Shaheed, Iman B; Abdel-Latif, Sally M

2011-01-01

281

Functional expression in insect cells of glycosylphosphatidylinositol-linked alkaline phosphatase from Aedes aegypti larval midgut: a Bacillus thuringiensis Cry4Ba toxin receptor.  

Science.gov (United States)

Bacillus thuringiensis produces insecticidal crystal (Cry) proteins which bind to cell surface receptors on the brush border membrane of susceptible midgut larvae. The toxin-receptor interaction generates pores in midgut epithelial cells resulting in cell lysis. Here, a cDNA encoding membrane-bound alkaline phosphatase from Aedes aegypti (Aa-mALP) midgut larvae, based on the sequence identity hit to Bombyx mori membrane-bound ALP, was amplified by RT-PCR and transiently expressed in Spodoptera frugiperda (Sf9) insect cells as a 58-kDa membrane-bound protein via the baculovirus expression system and confirmed by digestion with phosphatidylinositol-specific phospholipase C and LC-MS/MS analysis. Immunolocalization results showed that Cry4Ba is able to bind to only Sf9 cells-expressing Aa-mALP. Moreover, these cells were shown to undergo cell lysis in the presence of 100 ?g/ml trypsin-treated toxin. Finally, trypan blue exclusion assay also demonstrated an increase in cell death in recombinant cells treated with Cry4Ba. Overall results indicated that Aa-mALP protein was responsible for mediating Cry4Ba toxicity against Sf9 cells, suggesting its role as a receptor for Cry4Ba toxin in A. aegypti mosquito larvae. PMID:21146607

Dechklar, Manasave; Tiewsiri, Kasorn; Angsuthanasombat, Chanan; Pootanakit, Kusol

2011-03-01

282

Stable expression of the alkaline phosphatase marker gene by neural cells in culture and after transplantation into the CNS using cells derived from a transgenic rat.  

Science.gov (United States)

Multipotent stem cells and more developmentally restricted precursors have previously been isolated from the developing nervous system and their properties analyzed by culture assays in vitro and by transplantation in vivo. However, the variety of labeling techniques that have been used to identify grafted cells in vivo have been unsatisfactory. In this article we describe the characteristics of cells isolated from a transgenic rat in which the marker gene human placental alkaline phosphatase (hPAP) is linked to the ubiquitously active R26 gene promoter. We show that hPAP is readily detected in embryonic neuroepithelial stem cells, neuronal-restricted precursor cells, and glial-restricted precursor cells. Transgene expression is robust and can be detected by both immunocytochemistry and histochemistry. Furthermore, the levels of hPAP on the cell surface are sufficient for live cell labeling and fluorescence-activated cell sorting. Expression of hPAP is stable in isolated cells in culture and in cells transplanted into the spinal cord for at least 1 month. We submit that cells isolated from this transgenic rat will be valuable for studies of neural development and regeneration. PMID:11869033

Mujtaba, Tahmina; Han, Steve S W; Fischer, Itzhak; Sandgren, Eric P; Rao, Mahendra S

2002-03-01

283

Differences in growth and alkaline phosphatase activity between Microcystis aeruginosa and Chlorella pyrenoidosa in response to media with different organic phosphorus  

Directory of Open Access Journals (Sweden)

Full Text Available The growth of Microcystis aeruginosa and Chlorella pyrenoidosa in three dissolved organic phosphorus sources (glucose-1- phosphate, adenosine triphosphate, cyclic-adenosine monophosphate were studied in cultures separated by a dialysis membrane. Results showed that M. aeruginosa and C. pyrenoidosa could utilize those three forms of organic phosphorus, but their growth rates and cell abundances were low in comparison with those in the orthophosphate control. M. aeruginosa had a higher growth rate than C. pyrenoidosa in glucose-1-phosphate, and then became dominate in the separate cultures. In contrast, those two algal species didn’t show any significant differences in the growth rate and cell abundance in the medium with adenosine triphosphate and cyclicadenosine monophosphate. Alkaline phosphatase was an important enzyme for hydrolyzing glucose-1-phosphate, adenosine triphosphate and cyclic-adenosine monophosphate, the activity of which was positively correlated with the growth rate of algae. Considering the big proportion of glucose-1-phosphate in the Lake Taihu, the capability of M. aeruginosa to efficiently utilize this type of organic phosphorus source might be one of reason that why M. aeruginosa is the dominant species in this hyper-eutrophic lake.

Yang YU

2011-02-01

284

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

Science.gov (United States)

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

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

2014-01-01

285

Effects of dietary vitamin E on mucosal maltase and alkaline phosphatase enzyme activities and on the amount of mucosal malonyldialdehyde in broiler chickens  

Science.gov (United States)

The effects of dietary vitamin E levels on mucosal maltase and alkaline phosphatase (ALP) enzyme activities and on the amount of mucosal malonyldialdehyde (MDA) in broiler chickens were studied in the present study. One hundred and eighty of male day old broiler chicks (Ross 308 strain) were randomly assigned into five groups, each with three replicates and 12 chicks in each replicate. Chickens in group A were fed corn-soy- based diet, while those in groups B, C, D and E were fed the same diet with 20, 60, 180, and 540 mg kg-1 vitamin E supplement (d-alpha tocopherol), respectively. Six birds were randomly chosen from each group, and were euthanized on days 10, 21, 32, and 42 of age. One segment of small intestine outset was homogenized and mucosal ALP and maltase activity were measured. Moreover, mucosal lipid peroxidate amount was measured to reveal the impact of vitamin E on oxidative stress. Maltase activity was increased with the increase of vitamin E up to 60 mg kg-1 of diet while with further levels, it was decreased. Addition of 60 mg kg-1 of vitamin E to the diet significantly increased ALP enzyme activity (p ? 0.001). Addition of 540 mg kg-1 of vitamin E supplement to the diet led to the minimum amount of MDA at 32 days of age. It may be concluded that supplementation of broiler's diet with 60 mg kg-1 of vitamin E can increase mucosal maltase and ALP enzyme activity. PMID:25568675

Farrokhifar, Seyed Hamid; Ali Jafari, Ramezan; Erfani Majd, Naeem; Fatemi Tabatabaee, Seyed Reza; Mayahi, Mansour

2013-01-01

286

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

Science.gov (United States)

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

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

2014-01-01

287

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

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

Benjamin Nnamdi Okolonkwo

2013-01-01

288

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

DEFF Research Database (Denmark)

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

Rangnoi, Kuntalee; Jaruseranee, Nanthnit

2011-01-01

289

One-step detection of aflatoxin-B(1) using scFv-alkaline phosphatase-fusion selected from human phage display antibody library.  

Science.gov (United States)

A unique human phage display library was used to successfully generate a scFv to the highly carcinogenic toxin aflatoxin B1. Such an antibody has major potential applications in therapy and diagnostics. To further exploit its analytical capacity, the scFv was genetically fused to alkaline phosphatase, thereby generating a novel and highly sensitive self-indicating reagent. The performance of this reagent was further characterized, demonstrating its efficacy. The sensitivity of scFv-AP fusion was three-fold better than that of the scFv form. The ability of this human library to generate antibodies to a small hapten was clearly demonstrated and this is linked to its intrinsic diversity, which exceeds many existing conventional human libraries. Our results indicate that demography may influence the diversity of the repertoire of the library in terms of its capacity to generate antibodies to specific targets. Equally, the approach demonstrated should also be applicable for other haptens and larger antigens. PMID:21465334

Rangnoi, Kuntalee; Jaruseranee, Nanthnit; O'Kennedy, Richard; Pansri, Potjamas; Yamabhai, Montarop

2011-11-01

290

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

Science.gov (United States)

A rapid and sensitive direct competitive fluorescence enzyme immunoassay (dc-FEIA) for ochratoxin A (OTA) based on a nanobody (Nb)-alkaline phosphatase (AP) fusion protein was developed. The VHH (variable domain of heavy chain antibody) gene of Nb28 was subcloned into the expression vector pecan45 containing the AP double-mutant gene. The Nb28-AP construct was transformed into Escherichia coli BL21(DE3)plysS, and soluble expression in bacteria was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. Both the Nb properties and AP enzymatic activity were validated by colorimetric and fluorometric analysis. The 50% inhibitory concentration and the detection limit of the dc-FEIA were 0.13 and 0.04 ng/mL, respectively, with a linear range of 0.06-0.43 ng/mL. This assay was compared with LC-MS/MS, and the results indicated the reliability of Nb-AP fusion protein-based dc-FEIA for monitoring OTA contamination in cereal. PMID:25531426

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

2015-01-20

291

Sustained Osteomalacia of Long Bones Despite Major Improvement in Other Hypophosphatasia-Related Mineral Deficits in Tissue Nonspecific Alkaline Phosphatase/Nucleotide Pyrophosphatase Phosphodiesterase 1 Double-Deficient Mice  

OpenAIRE

We have shown previously that the hypomineralization defects of the calvarium and vertebrae of tissue nonspecific alkaline phosphatase (TNAP)-deficient (Akp2?/?) hypophosphatasia mice are rescued by simultaneous deletion of the Enpp1 gene, which encodes nucleotide pyrophosphatase phosphodiesterase 1 (NPP1). Conversely, the hyperossification in the vertebral apophyses typical of Enpp1?/? mice is corrected in [Akp2?/?; Enpp1?/?] double-knockout mice. Here we have examined the ap...

Anderson, H. Clarke; Harmey, Dympna; Camacho, Nancy P.; Garimella, Rama; Sipe, Joseph B.; Tague, Sarah; Bi, Xiaohong; Johnson, Kristen; Terkeltaub, Robert; Milla?n, Jose? Luis

2005-01-01

292

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

Directory of Open Access Journals (Sweden)

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

Parker Bruce R

2009-07-01

293

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

Science.gov (United States)

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

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

2010-09-01

294

Therapeutic potentials of ecto-nucleoside triphosphate diphosphohydrolase, ecto-nucleotide pyrophosphatase/phosphodiesterase, ecto-5'-nucleotidase, and alkaline phosphatase inhibitors.  

Science.gov (United States)

The modulatory role of extracellular nucleotides and adenosine in relevance to purinergic cell signaling mechanisms has long been known and is an object of much research worldwide. These extracellular nucleotides are released by a variety of cell types either innately or as a response to patho-physiological stress or injury. A variety of surface-located ecto-nucleotidases (of four major types; nucleoside triphosphate diphosphohydrolases or NTPDases, nucleotide pyrophosphatase/phosphodiesterases or NPPs, alkaline phosphatases APs or ALPs, and ecto-5'-nucleotidase or e5NT) are responsible for meticulously controlling the availability of these important signaling molecules (at their respective receptors) in extracellular environment and are therefore crucial for maintaining the integrity of normal cell functioning. Overexpression of many of these ubiquitous ecto-enzymes has been implicated in a variety of disorders including cell adhesion, activation, proliferation, apoptosis, and degenerative neurological and immunological responses. Selective inhibition of these ecto-enzymes is an area that is currently being explored with great interest and hopes remain high that development of selective ecto-nucleotidase inhibitors will prove to have many beneficial therapeutic implications. The aim of this review is to emphasize and focus on recent developments made in the field of inhibitors of ecto-nucleotidases and to highlight their structure activity relationships wherever possible. Most recent and significant advances in field of NTPDase, NPP, AP, and e5NT inhibitors is being discussed in detail in anticipation of providing prolific leads and relevant background for research groups interested in synthesis of selective ecto-nucleotidase inhibitors. PMID:24115166

al-Rashida, Mariya; Iqbal, Jamshed

2014-07-01

295

Signal-on electrochemical immunoassay for APE1 using ionic liquid doped Au nanoparticle/graphene as a nanocarrier and alkaline phosphatase as enhancer.  

Science.gov (United States)

In this paper, the Au nanoparticles decorated graphene nanosheets (AuNPs/Gr) were prepared as nanocarriers using ionic liquid (IL) as linker reagent. Then the alkaline phosphatase (ALP) and the ferrocene tagged detection antibodies (Fc-Ab2) were loaded on the IL doped AuNPs/Gr as a trace label for ultrasensitive measurements of human apurinic/apyrimidinic endonuclease 1 (APE1), which is a multifunctional protein in the DNA base excision repair pathway relating to various types of cancer. Several labeling protocols were investigated for the determination of the APE1 protein concentration and improved analytical features were obtained with the proposed carriers of IL doped AuNPs/Gr which were labeled with Fc-Ab2 and ALP (ALP/Fc-Ab2/AuNPs/IL/Gr). The reason may be that the IL doped AuNPs/Gr carriers (AuNPs/IL/Gr) could not only enhance the immobilized amount of ALP and Fc-Ab2, but also promote the electron transfer rate. Thus, through the specific recognition of antigen-antibody, numerous ALP/Fc-Ab2/AuNPs/IL/Gr, which are captured onto every single immunocomplex, could further catalyze the ascorbic acid 2-phosphate (AA-p) reaction to amplify the electrochemical signal. Transmission electron microscopy (TEM) images of the AuNPs/IL/Gr nanocomposites revealed the formation of a functionalized surface network structure. The resulting immunosensor exhibited a linear response to APE1 in the concentration range of 0.1-80 pg mL(-1) with a detection limit of 0.04 pg mL(-1), indicating potential applications in clinical diagnostics. PMID:25356934

Zhong, Zhaoyang; Li, Mengxia; Qing, Yi; Dai, Nan; Guan, Wei; Liang, Wei; Wang, Dong

2014-12-21

296

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

Science.gov (United States)

An ultrasensitive sandwich-type analysis method has been initially developed for probing low-level free microRNAs (miRNAs) in blood by a maximal signal amplification protocol of catalytic silver deposition. Gold nanoclusters (AuNCs) were first synthesized and in-site incorporated into alkaline phosphatase (ALP) to form the ALP-AuNCs. Unexpectedly, the so incorporated AuNCs could dramatically enhance the catalysis activities of ALP-AuNCs versus native ALP. A sandwiched hybridization protocol was then proposed using ALP-AuNCs as the catalytic labels of the DNA detection probes for targeting miRNAs that were magnetically caught from blood samples by DNA capture probes, followed by the catalytic ligation of two DNA probes complementary to the targets. Herein, the ALP-AuNC labels could act as the bicatalysts separately in the ALP-catalyzed substrate dephosphorylation reaction and the AuNCs-accelerated silver deposition reaction. The signal amplification of ALP-AuNCs-catalyzed silver deposition was thereby maximized to be measured by the electrochemical outputs. The developed electroanalysis strategy could allow for the ultrasensitive detection of free miRNAs in blood with the detection limit as low as 21.5 aM, including the accurate identification of single-base mutant levels in miRNAs. Such a sandwich-type analysis method may circumvent the bottlenecks of the current detection techniques in probing short-chain miRNAs. It would be tailored as an ultrasensitive detection candidate for low-level free miRNAs in blood toward the diagnosis of cancer and the warning or monitoring of cancer metastasis in the clinical laboratory. PMID:25242013

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

2014-10-21

297

Effects of dietary vitamin E on mucosal maltase and alkaline phosphatase enzyme activities and on the amount of mucosal malonyldialdehyde in broiler chickens  

Directory of Open Access Journals (Sweden)

Full Text Available The effects of dietary vitamin E levels on mucosal maltase and alkaline phosphatase (ALP enzyme activities and on the amount of mucosal malonyldialdehyde (MDA in broiler chickens were studied in the present study. One hundred and eighty of day old male broiler chicks (Ross 308 strain were randomly assigned into five groups, each with three replicates and 12 chicks in each replicate. Chickens in group A were fed corn-soy- based diet, while those in groups B, C, D and E were fed the same diet with 20, 60, 180, and 540 mg kg-1 vitamin E supplement (d-alpha tocopherol, respectively. Six birds were randomly chosen from each group, and were euthanized on days 10, 21, 32, and 42 of age. One segment of small intestine outset was homo-genized and mucosal ALP and maltase activity were measured. Moreover, mucosal lipid peroxidate amount was measured to reveal the impact of vitamin E on oxidative stress. Maltase activity was increased with the increase of vitamin E up to 60 mg kg-1 of diet while with further levels, it was decreased. Addition of 60 mg kg-1 of vitamin E to the diet significantly increased ALP enzyme activity (p ? 0.001. Addition of 540 mg kg-1 of vitamin E supplement to the diet led to the minimum amount of MDA at 32 days of age. It may be concluded that supplementation of broiler's diet with 60 mg kg-1of vitamin E can increase mucosal maltase and ALP enzyme activity.

Seyed Hamid Farrokhifar

2014-12-01

298

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

Science.gov (United States)

Alkaline phosphatase (ALP) is a ubiquitous enzyme in milk with time-temperature destruction similar to that of certain pathogens destroyed in pasteurization. Measurement of ALP to indicate proper pasteurization is a common practice. Recently the public health level for ALP was decreased to 350 mU/L, a level below the sensitivity of older colorimetric ALP methods. This study was conducted within the structure of the International Dairy Federation and the International Organization for Standardization to evaluate the reproducibility of the chemiluminescence method (Charm PasLite) for ALP at 50, 100, 350, and 500 mU/L in whole milk of multiple species to meet new regulations in the United States and proposed regulations in the European Union (EU). Fifteen laboratories from 8 countries evaluated bovine, goat, sheep, and buffalo milk, bovine skim milk, 20% fat cream, and 2% fat chocolate milk. At ALP levels of 350 and 500 mU/L, the average relative standard deviation for repeatability (RSDr) was 7.5%, and the average relative standard deviation of reproducibility was (RSDR) 15%. For ALP at 100 and 50 mU/L, the average RSDr values were 10.5 and 12.6%, respectively, and the average RSDR values were 18 and 25%, respectively. The limit of detection was 20 mU/L. Results are comparable to those obtained with other enzymatic photo-activated system methods such as the fluorometric method. Results indicate that the method is suitable for measuring ALP in the milk of multiple species and in dairy drinks at U.S. and proposed EU levels. PMID:16915846

Salter, Robert S; Fitchen, John

2006-01-01

299

Chicken single-chain antibody fused to alkaline phosphatase detects Aspergillus pathogens and their presence in natural samples by direct sandwich enzyme-linked immunosorbent assay.  

Science.gov (United States)

A sensitive and specific analytical method to detect ubiquitous aflatoxigenic Aspergillus pathogens is essential for monitoring and controlling aflatoxins. Four highly reactive chicken single-chain variable fragments (scFvs) against soluble cell wall proteins (SCWPs) from Aspergillus flavus were isolated by phage display. The scFv antibody AfSA4 displayed the highest activity toward both A. flavus and A. parasiticus and specifically recognized a surface target of their cell walls as revealed by immunofluorescence localization. Molecular modeling revealed a unique compact motif on the antibody surface mainly involving L-CDR2 and H-CDR3. As measured by surface plasmon resonance, AfSA4 fused to alkaline phosphatase had a higher binding capability and 6-fold higher affinity compared with AfSA4 alone. Immunoblot analyses showed that the fusion had good binding capacity to SCWP components from the two fungal species. Direct sandwich enzyme-linked immunosorbent assays with mouse antiaspergillus monoclonal antibody mAb2A8 generated in parallel as a capture antibody revealed that the detection limit of the two fungi was as low as 10(-3) ?g/mL, 1000-fold more sensitive than that reported previously (1 ?g/mL). The fusion protein was able to detect fungal concentrations below 1 ?g/g of maize and peanut grains in both artificially and naturally contaminated samples, with at least 10-fold more sensitivity than that reported (10 ?g/g) thus far. Thus, the fusion can be applied in rapid, simple, and specific diagnosis of Aspergillus contamination in field and stored food/feed commodities. PMID:24128348

Xue, Sheng; Li, He-Ping; Zhang, Jing-Bo; Liu, Jin-Long; Hu, Zu-Quan; Gong, An-Dong; Huang, Tao; Liao, Yu-Cai

2013-11-19

300

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

Scientific Electronic Library Online (English)

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

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

2005-10-01

301

Comparison of micro- vs. nanostructured colloidal gelatin gels for sustained delivery of osteogenic proteins: Bone morphogenetic protein-2 and alkaline phosphatase.  

Science.gov (United States)

Colloidal gels have recently emerged as a promising new class of materials for regenerative medicine by employing micro- and nanospheres as building blocks to assemble into integral scaffolds. To this end, physically crosslinked particulate networks are formed that are injectable yet cohesive. By varying the physicochemical properties of different particle populations, the suitability of colloidal gels for programmed delivery of multiple therapeutic proteins is superior over conventional monolithic gels that lack this strong capacity for controlled drug release. Colloidal gels made of biodegradable polymer micro- or nanospheres have been widely investigated over the past few years, but a direct comparison between micro- vs. nanostructured colloidal gels has not been made yet. Therefore, the current study has compared the viscoelastic properties and capacity for drug release of colloidal gels made of oppositely charged gelatin microspheres vs. nanospheres. Viscoelastic properties of the colloidal gelatin gels were characterized by rheology and simple injectability tests, and in vitro release of two selected osteogenic proteins (i.e. bone morphogenetic protein-2 (BMP-2) and alkaline phosphatase (ALP)) from the colloidal gelatin gels was evaluated using radiolabeled BMP-2 and ALP. Nanostructured colloidal gelatin gels displayed superior viscoelastic properties over microsphere-based gels in terms of elasticity, injectability, structural integrity, and self-healing behavior upon severe network destruction. In contrast, microstructured colloidal gelatin gels exhibited poor gel strength and integrity, unfavorable injectability, and did not recover after shearing, resulting from the poor gel cohesion due to insufficiently strong interparticle forces. Regarding the capacity for drug delivery, sustained growth factor (BMP-2) release was obtained for both micro- and nanosphere-based gels, the kinetics of which were mainly depending on the particle size of gelatin spheres with the same crosslinking density. Therefore, the optimal gelatin carrier for drug delivery in terms of particle size and crosslinking density still needs to be established for specific clinical indications that require either short-term or long-term release. It can be concluded that nanostructured colloidal gelatin gels show great potential for sustained delivery of therapeutic proteins, whereas microstructured colloidal gelatin gels are not sufficiently cohesive as injectables for biomedical applications. PMID:22922022

Wang, Huanan; Boerman, Otto C; Sariibrahimoglu, Kemal; Li, Yubao; Jansen, John A; Leeuwenburgh, Sander C G

2012-11-01

302

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

International Nuclear Information System (INIS)

Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: ? The scFv-AP fusion protein against ractopamine (RAC) was produced. ? A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. ? The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. ? Recovery tests from pork samples were studied. ? Good accuracy was obtained. - Abstract: A rapid, sensitive chemiluminescent enzyme immunoassay (CLEIA) for ractopamine (RAC) based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein was developed. The scFv gene was prepared by cloning the heavy- and light-chain variable region genes (VH and VL) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling VH and VL genes with a linker by means of splicing overlap extension polymerase chain reaction. The resulting scFv gene was inserted into the expression vector pLIP6/GN containing AP to produce the fusion protein in Escherichia coli strain BL21. The purified scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) protocol for detection of RAC. The average concentration required for 50% inhibition of binding and the limit of detection of the assay were 0.25 ± 0.03 and 0.02 ± 0.004 ng mL?1, respectively, and the linear response range extended from 0.05 to 1.45 ng mL?1. The assay was 10 ti/sup>. The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS). The results showed a good correlation between the data of dc-CLEIA and HPLC–MS (R2 > 0.99), indicating that the assay was an efficient analytical method for monitoring food safety.

303

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

Energy Technology Data Exchange (ETDEWEB)

Graphical abstract: Detection model of dc-CLEIA based on anti-RAC scFv-AP fusion protein. Highlights: Black-Right-Pointing-Pointer The scFv-AP fusion protein against ractopamine (RAC) was produced. Black-Right-Pointing-Pointer A dc-CLEIA for RAC was developed based on the purified scFv-AP fusion protein. Black-Right-Pointing-Pointer The sensitivity of dc-CLEIA was 10 times as sensitive as dc-ELISA for RAC. Black-Right-Pointing-Pointer Recovery tests from pork samples were studied. Black-Right-Pointing-Pointer Good accuracy was obtained. - Abstract: A rapid, sensitive chemiluminescent enzyme immunoassay (CLEIA) for ractopamine (RAC) based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein was developed. The scFv gene was prepared by cloning the heavy- and light-chain variable region genes (V{sub H} and V{sub L}) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling V{sub H} and V{sub L} genes with a linker by means of splicing overlap extension polymerase chain reaction. The resulting scFv gene was inserted into the expression vector pLIP6/GN containing AP to produce the fusion protein in Escherichia coli strain BL21. The purified scFv-AP fusion protein was used to develop a direct competitive CLEIA (dcCLEIA) protocol for detection of RAC. The average concentration required for 50% inhibition of binding and the limit of detection of the assay were 0.25 {+-} 0.03 and 0.02 {+-} 0.004 ng mL{sup -1}, respectively, and the linear response range extended from 0.05 to 1.45 ng mL{sup -1}. The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The results showed a good correlation between the data of dc-CLEIA and HPLC-MS (R{sup 2} > 0.99), indicating that the assay was an efficient analytical method for monitoring food safety.

Dong Jiexian; Li Zhenfeng; Lei Hongtao; Sun Yuanming [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Ducancel, Frederic [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Xu Zhenlin [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Boulain, Jean-Claude [CEA, iBiTec-S, Service de Pharmacologie et d' Immnoanalyse (SPI), CEA Saclay, F-91191 Gif sur Yvette (France); Yang Jinyi; Shen Yudong [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China); Wang Hong, E-mail: gzwhongd@63.com [Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642 (China)

2012-07-29

304

Fosfatasa alcalina intestinal: una enzima con propiedades antiinflamatorias / Intestinal alkaline phosphatase: an enzyme with anti-inflammatory properties / Fosfatasse alcalina intestinal: uma enzima com propriedades anti-inflamatórias  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in portuguese Resumo Uma das principais funções da Fosfatasse Alcalina Intestinal (FAI) é a detoxificação dos lipopolissacarídeos (LPS) bacterianos para controlar a inflamação intestinal. Recentes publicações indicam que a FAI participa na detoxificação de outros compostos bacterianos (flagelina e motivos CpG do [...] DNA) e de muitos nucleotídeos libres (ATP, UDP). A FAI está involucrada de forma direita na recuperação tissular da inflamação pela Resolvina E1 (RvE1). A ação antiinflamatória da FAI melhora indiretamente a função da barreira intestinal e impacta a diversidade e a composição da microbiota. Diversas doenças intestinais, incluindo enterocolitis necrótica, doença celíaca e a inflamação crônica intestinal (inflammatory bowel disease, IBD) estão relacionados com diminuições na expressão e atividade da FAI. De outro jeito, uma elevada atividade da FAI no cólon é sinônimo de processos inflamatórios, devido a elevada concentração da isoforma tissular da Fosfatasse Alcalina não especifica (FANE), e a infiltração tissular pelos neutrófilos (que também contém FANE). A administração exógena da FAI reduz a inflamação intestinal/sistêmica (dependendo da via de administração) incluindo uns poucos testes no homem. Em conclusão, a homeostase intestinal e a preservação da saúde dependem em grande medida da capacidade da FAI para detoxificar os LPS e suprimir a inflamação metabólica induzida por estes. Embora, é preciso realizar pesquisas bem feitas sobre como os costumes alimentares podem modificar a detoxificação dos diferentes compostos proinflamatórios bacterianos e maximizar a Abstract in spanish Resumen Una de las principales funciones de la Fosfatasa Alcalina Intestinal (FAI) es la detoxificación de los lipopolisacáridos (LPS) bacterianos para controlar la inflamación intestinal. Recientes publicaciones indican que FAI participa en la detoxificación de otros compuestos bacterianos (flageli [...] na y motivos CpG de DNA) y de muchos nucleótidos libres (ATP, UDP). FAI está involucrada de manera directa en la recuperación tisular de la inflamación por la Resolvina E1. La acción antiinflamatoria de FAI mejora indirectamente la función de la barrera intestinal e impacta la diversidad y la composición de la microbiota. Diversas enfermedades intestinales, incluyendo enterocolitis necrótica, enfermedad celíaca y la inflamación crónica intestinal (o inflammatory bowel disease, IBD) están relacionadas con disminuciones en la expresión y actividad de FAI. Por otro lado, una elevada actividad de FAI en colon es sinónimo de procesos inflamatorios, debido a la elevada concentración de la isoforma tisular de Fosfatasa Alcalina no específica (FANE), y a la infiltración tisular por los neutrófilos (que también contienen FANE). En algunos ensayos en humanos se ha observado que la administración exógena de FAI reduce la inflamación intestinal/sistémica (dependiendo de la vía de administración). En conclusión, la homeóstasis intestinal y la preservación de la salud dependen en gran medida de la capacidad de FAI para detoxificar los LPS y suprimir la inflamación metabólica inducida por estos. Sin embargo, es necesario realizar investigaciones a fondo sobre como los hábitos alimenticios pueden modificar la detoxificación de los diferentes compuestos proinflamatorios bacterianos y maximizar la actividad de FAI Abstract in english Abstract One of the main functions of Intestinal Alkaline Phosphatase (FAI) is to detoxify bacterial lipopolysaccharides (LPS) to control intestinal inflammation. Recent data indicate that FAI participates in the detoxification of other bacterial compounds (flagellin and DNA CpG motifs) and many fre [...] e nucleotides (ATP, UDP). FAI is directly involved in the resolution of tissue inflammation mediated by Resolvin E1. The anti-inflammatory action of FAI indirectly improves the intestinal barrier function and affects the diversity of microbiota. Var

Jean-Paul, Lallès; Jaime Parra, Suescún.

2014-06-01

305

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

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Full Text Available Biliary complications often lead to acute and chronic liver injury after orthotopic liver transplantation (OLT. Bile composition and secretion depend on the integrated action of all the components of the biliary tree, starting from hepatocytes. Fatty livers are often discarded as grafts for OLT, since they are extremely vulnerable to conventional cold storage (CS. However, the insufficiency of donors has stimulated research to improve the usage of such marginal organs as well as grafts. Our group has recently developed a machine perfusion system at subnormothermic temperature (20°C; MP20 that allows a marked improvement in preservation of fatty and even of normal rat livers as compared with CS. We sought to evaluate the response of the biliary tree of fatty liver to MP20, and a suitable marker was essential to this purpose. Alkaline phosphatase (AlkP, EC 3.1.3.1, frequently used as marker of membrane transport in hepatocytes and bile ducts, was our first choice. Since no histochemical data were available on AlkP distribution and activity in fatty liver, we have first settled to investigate AlkP activity in the steatotic liver of fatty Zucker rats (fa/fa, using as controls lean Zucker (fa/+ and normal Wistar rats. The AlkP reaction in Wistar rats was in accordance with the existing data and, in particular, was present in bile canaliculi of hepatocytes in the periportal region and midzone, in the canals of Hering and in small bile ducts but not in large bile ducts. In lean ZR liver the AlkP reaction in Hering canals and small bile ducts was similar to Wistar rat liver but hepatocytes had lower canalicular activity and besides presented moderate basolateral reaction. The difference between lean Zucker and Wistar rats, both phenotypically normal animals, could be related to the fact that lean Zucker rats are genotypically heterozygous for a recessive mutated allele. In fatty liver, the activity in ductules and small bile ducts was unchanged, but most hepatocytes were devoid of AlkP activity with the exception of clusters of macrosteatotic hepatocytes in the mid-zone, where the reaction was intense in basolateral domains and in distorted canaliculi, a typical pattern of cholestasis. The interpretation of these data was hindered by the fact that the physiological role of AlkP is still under debate. In the present study, the various functions proposed for the role of the enzyme in bile canaliculi and in cholangiocytes are reviewed. Independently of the AlkP role, our data suggest that AlkP does not seem to be a reliable marker to study the initial step of bile production during OLT of fatty livers, but may still be used to investigate the behaviour of bile ductules and small bile ducts.

V. Bertone

2011-02-01

306

iagnostic accuracy study comparing total alkaline phosphatase with intact parathyroid hormone 1-84 for the diagnosis of high turnover renal osteodystrophy in chronic renal failure on hemodialysis  

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Full Text Available INTRODUCTION High turnover renal osteodystrophy (HTRO is a highly prevalent complication in patients with chronic kidney disease and mineral bone disease (CKD-MBD, causing pain and significant fracture-associated morbidity and mortality. The diagnostic gold standard test is bone biopsy but there are other, more widely available screening tests such as 1-84 intact parathormone (1-84 iPTH and nonspecific markers such as total alkaline phosphatase (tALP. PURPOSE To determine the diagnostic value (ROC curve, predictive values and likelihood ratios of 1-84 iPTH and tALP for HTRO screening. METHODS A diagnostic accuracy study was performed on a sample of CKD-MDB patients, grouping them according to bone biopsy results and analyzing the results of the diagnostic tests as descriptive variables. RESULTS The study group comprised 188 patients with CKD-MDB, 36 of which had biopsy-confirmed HTRO (19.15%. The average age was 50.2 years in the biopsy group, and 53.4 years in the non-biopsy group (p=0.2385, most were male (63.8% and diabetic (80.5%. The mean time in dialysis was 5.02 years in the biopsy group, and 2.61 years for the non-biopsy group (p<0.001. The mean Kt/V was 1.44 in the biopsy group, and 1.40 in the non-biopsy group (p=0.5354. The mean tALP was 398.02 IU/L in the group with HTRO versus 141.76 IU/L in the group without HTRO (p<0.001. The best cut-off value for tALP was 300-350 IU/L with a near 80% post-test probability, but also with a 15-20% probability for HTRO if the test is negative. The mean 1-84 iPTH was 1248.01 pg/ml in the group with HTRO versus 350.76 pg/ml in the group without HTRO (p<0.001. The 1-84 iPTH cut-off reference value of 300 pg/ml was associated with a post-test probability of 30% for HTRO diagnosis and had a lower overall performance. The best cut-off value for iPTH 1-84 was 600 pg/ml with a post-test probability for HTRO of 70% if positive and less than 5% if the test results are negative. DISCUSSION Both markers show good correlation with bone biopsy findings. tALP elevation detects presence of HTRO in selected patients but does not rule it out. tALP does not perform as well as 1-84 iPTH as a screening test for HTRO.

Andrés Marcelo Rojas González

2014-09-01

307

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Argentina | Language: Spanish Abstract in spanish La transformación de los compuestos de fósforo orgánico (Po) a fósforo inorgánico (Pi) soluble, es denominada mineralización y es llevada a cabo por un grupo de enzimas conocidas como fosfatasas. En este trabajo, se estudió la actividad fosfatasa de cinco lotes de la región pampeana norte del área s [...] ojera argentina, mediante: la evaluación de la actividad fosfatasa del suelo, y el recuento de las comunidades bacterianas y fúngicas con esa actividad, y de esta manera se obtuvo información sobre el potencial de los mismos para movilizar el Po. Se determinó el número de bacterias aeróbicas heterotróficas cultivables (BAHC) así como el de hongos cultivables (HC), además del número de productores de fosfatasas ácidas y alcalinas. El número de bacterias con actividad fosfatasa fue en promedio 6,85 10(5) UFC g-1 de suelo para la fosfatasa ácida; mientras que para la fosfatasa alcalina fue en promedio 5,80 10(5) UFC g-1 de suelo. En cambio, el valor medio de hongos con actividad fosfatasa ácida fue 1,78 10³ UFC g-1 de suelo y para la enzima alcalina 1,77 10³ UFC g-1 de suelo. No se encontraron diferencias estadísticamente significativas entre el número de bacterias y el de hongos con fosfatasa ácida o alcalina entre los distintos suelos. Por otro lado, el nivel de actividad de la fosfatasa alcalina osciló entre 5,72 y 15,5 mg p-nitrofenol kg-1 suelo h-1, mientras la ácida varió entre 27,4 y 105 mg p-nitrofenol kg-1 suelo h-1. Se encontraron diferencias significativas en la actividad enzimática entre los cinco suelos, siendo mayor la actividad ácida que la alcalina. Los resultados de este trabajo demostraron que los suelos estudiados presentan una actividad mineralizadora de fuentes de Po que coincide con otros trabajos de suelos cultivados y que el recuento de los organismos productores de fosfatasas complementa la información obtenida a partir de la determinación de la actividad fosfatasa del suelo. Mediante la utilización de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po. Abstract in english Transformation of organic phosphorus (Po) into soluble inorganic phosphorus (Pi) is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating [...] the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB) and cultivated fungi (CF) was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5) CFU g-1 soil, while alkaline activity was 5.80 10(5) CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5) mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

Leticia Andrea, Fernández; Marcelo Antonio, Sagardoy; Marisa Anahí, Gómez.

2008-07-01

308

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

Directory of Open Access Journals (Sweden)

Full Text Available La transformación de los compuestos de fósforo orgánico (Po a fósforo inorgánico (Pi soluble, es denominada mineralización y es llevada a cabo por un grupo de enzimas conocidas como fosfatasas. En este trabajo, se estudió la actividad fosfatasa de cinco lotes de la región pampeana norte del área sojera argentina, mediante: la evaluación de la actividad fosfatasa del suelo, y el recuento de las comunidades bacterianas y fúngicas con esa actividad, y de esta manera se obtuvo información sobre el potencial de los mismos para movilizar el Po. Se determinó el número de bacterias aeróbicas heterotróficas cultivables (BAHC así como el de hongos cultivables (HC, además del número de productores de fosfatasas ácidas y alcalinas. El número de bacterias con actividad fosfatasa fue en promedio 6,85 10(5 UFC g-1 de suelo para la fosfatasa ácida; mientras que para la fosfatasa alcalina fue en promedio 5,80 10(5 UFC g-1 de suelo. En cambio, el valor medio de hongos con actividad fosfatasa ácida fue 1,78 10³ UFC g-1 de suelo y para la enzima alcalina 1,77 10³ UFC g-1 de suelo. No se encontraron diferencias estadísticamente significativas entre el número de bacterias y el de hongos con fosfatasa ácida o alcalina entre los distintos suelos. Por otro lado, el nivel de actividad de la fosfatasa alcalina osciló entre 5,72 y 15,5 mg p-nitrofenol kg-1 suelo h-1, mientras la ácida varió entre 27,4 y 105 mg p-nitrofenol kg-1 suelo h-1. Se encontraron diferencias significativas en la actividad enzimática entre los cinco suelos, siendo mayor la actividad ácida que la alcalina. Los resultados de este trabajo demostraron que los suelos estudiados presentan una actividad mineralizadora de fuentes de Po que coincide con otros trabajos de suelos cultivados y que el recuento de los organismos productores de fosfatasas complementa la información obtenida a partir de la determinación de la actividad fosfatasa del suelo. Mediante la utilización de ambos métodos, es posible estudiar la fosfatasa ácida y alcalina de un suelo y obtener información sobre el potencial del mismo para movilizar Po.Transformation of organic phosphorus (Po into soluble inorganic phosphorus (Pi is called mineralization and is carried out by phosphatase enzymes. The present research focuses on the study of the phosphatase activity of five soils from the soybean area of the Northern Pampean region, by evaluating the phosphatase activity in soil samples and the number of bacteria and fungi with that activity. Soil samples were collected and the total number and phosphatase activity of cultivated heterotrophic aerobic bacteria (CHAB and cultivated fungi (CF was assessed. No significant differences were observed in the numbers of CHAB and CH between the studied soils. The number of bacteria with acid phosphatase activity was 6.85 10(5 CFU g-1 soil, while alkaline activity was 5.80 10(5 CFU g-1 soil. In contrast, the number of fungi with acid phosphatase activity was 1.78 10³ CFU g-1 soil and with alkaline activity was 1.77 10³ CFU g-1 soil. No significant differences were observed in the number of bacteria and fungi with both enzymes. However, acid activity was higher than alkaline activity in soil samples. Alkaline phosphatase activity ranged from 5.72 to 15.5 mg p- nitrofenol kg-1 soil h-1 while acid activity varied from 27.4 to 10(5 mg p-nitrofenol kg-1 soil h-1. There were significant differences in phosphatase activity between the soybean soils. Our results show that the mineralization activities of Po sources are in agreement with other cultivated soils. On the other hand, the number of bacteria and fungi complements the information on soil phosphatase activity. Clearly, both methods allow the study of alkaline and acid phosphatase activity in soil and give information about the soil potential to mobilize Po.

Leticia Andrea Fernández

2008-07-01

309

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

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

M.A. Geidam

2007-01-01

310

Application of Scharer's quantitative method for the determination of residual alkaline phosphatase activity in standard Minas / Aplicação do método modificado de Scharer para a determinação quantitativa da atividade de fosfatase alcalina residual em queijo minas padrão  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese A pasteurização do leite é um ponto crítico na indústria de laticínios, e falhas nessa etapa comprometem a segurança do produto. O método enzimático de Scharer é tradicionalmente utilizado na verificação da eficiência da pasteurização e baseia-se na pesquisa da atividade de fosfatase alcalina residu [...] al em leite. Embora vários métodos estejam disponíveis para avaliar a eficiência da pasteurização, há um número reduzido de dados publicados baseados na quantificação da atividade da fosfatase alcalina em queijo. Neste estudo, o método modificado de Scharer foi utilizado para determinar os níveis de fosfatase alcalina residual em queijo minas padrão, antes e após 20 dias de maturação. Os queijos foram feitos com leite cru ou com leite pasteurizado com adição de diferentes concentrações de leite cru (0, 0,05%, 0,10%, 0,20% e 0,50%). Nas amostras de queijo fresco, o método apresentou sensibilidade apenas com 0,50% de adição de leite cru ao leite pasteurizado utilizado na fabricação de queijo. Em níveis de adição de até 0,20% de leite cru no leite pasteurizado, as concentrações de fenol se mostraram inferiores a 1?g de fenol/g de produto lácteo, que é o valor preconizado como indicador de pasteurização adequada. Abstract in english Milk pasteurization is a critical issue in the dairy industry, and failures in this process can affect final product safety. Scharer's enzymatic method is still traditionally used to verify pasteurization efficiency compliance, and it is based on screening for residual alkaline phosphatase in milk. [...] Although several methods are used to quantify enzymatic activity to assess milk pasteurization efficiency, there is a small amount of published data regarding the use of these methods to quantify alkaline phosphatase in cheese. In this study, the Scharer's modified method was used to determine the levels of residual alkaline phosphatase in standard minas cheese, before and after 20 days of ripening. The cheeses were made using raw or pasteurized milk with the addition of different concentrations of raw milk (0; 0.05%; 0.10%; 0.20%; and 0.50%). In the fresh cheese samples, the method showed a sensitivity of only 0.50% with the addition of raw milk to the pasteurized milk used to make cheese. In addition, levels of up 0.20% of raw milk in pasteurized milk, the concentrations of phenol was inferior to 1?g phenol/g of dairy product which is the preconized indicator value for adequate pasteurization.

C.F., Soares; L.M., Fonseca; M.O., Leite; M.C.P.P., Oliveira.

1223-12-01

311

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

Science.gov (United States)

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

312

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

Scientific Electronic Library Online (English)

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

Monica S., Ferreira-Nozawa; Sérgio R., Nozawa; Nilce M., Martinez-Rossi; Antonio, Rossi.

2003-06-01

313

Protein Phosphatases  

Science.gov (United States)

This Teaching Resource provides lecture notes and slides for a class covering the structure and function of protein phosphatases and is part of the course "Cell Signaling Systems: A Course for Graduate Students." The lecture begins with a discussion of the importance of phosphatases in physiology, recognized by the award of a Nobel Prize in 1992, and then proceeds to describe the two types of protein phosphatases: serine/threonine and tyrosine phosphatases. The information covered includes the structure, regulation, and substrate specificity of protein phosphatases, with an emphasis on their importance in disease and clinical settings.

Stephen R. Salton (Mount Sinai School of Medicine; Department of Neuroscience REV)

2005-03-01

314

A study on serum enzyme levels in various liver diseases  

OpenAIRE

Patients with chronic liver diseases are asymptomatic or have only vague non-specific symptoms. Effective medical treatments for chronic liver disease (before cirrhosis is established) are becoming increasingly available and since abnormal LFTs may be the only indication of these diseases. Aims: Enzymes study of various liver diseases. Discussion: serum Alkaline phosphatase (ALP), Gamma Glutamyl transferase (Gamma GT), Alanine and Aspartate amino transferases were estimated in viral Hepatiti...

Salma Mahaboob R, Jayarami Reddy U.

2013-01-01

315

Thyroxine is the serum factor that regulates morphogenesis of columnar cartilage from isolated chondrocytes in chemically defined medium  

OpenAIRE

Epiphyseal chondrocytes cultured in a medium containing 10% serum may be maintained as three dimensional aggregates and differentiate terminally into hypertrophic cells. There is an attendant expression of genes encoding type X collagen and high levels of alkaline phosphatase activity. Manipulation of the serum concentration to optimal levels of 0.1 or 0.01% in this chondrocyte pellet culture system results in formation of features of developing cartilage architecture which have been observed...

1994-01-01

316

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

OpenAIRE

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

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

2004-01-01

317

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

Science.gov (United States)

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

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

2015-01-01

318

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

Scientific Electronic Library Online (English)

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

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

2001-04-01

319

Maternal Antibiotic-Induced Early Changes in Microbial Colonization Selectively Modulate Colonic Permeability and Inducible Heat Shock Proteins, and Digesta Concentrations of Alkaline Phosphatase and TLR-Stimulants in Swine Offspring.  

Science.gov (United States)

Elevated intake of high energy diets is a risk factor for the development of metabolic diseases and obesity. High fat diets cause alterations in colonic microbiota composition and increase gut permeability to bacterial lipopolysaccharide, and subsequent low-grade chronic inflammation in mice. Chronic inflammatory bowel diseases are increasing worldwide and may involve alterations in microbiota-host dialog. Metabolic disorders appearing in later life are also suspected to reflect changes in early programming. However, how the latter affects the colon remains poorly studied. Here, we hypothesized that various components of colonic physiology, including permeability, ion exchange and protective inducible heat shock proteins (HSP) are influenced in the short- and long-terms by early disturbances in microbial colonization. The hypothesis was tested in a swine model. Offspring were born to control mothers (n = 12) or mothers treated with the antibiotic (ATB) amoxicillin around parturition (n = 11). Offspring were slaughtered between 14 and 42 days of age to study short-term effects. For long-term effects, young adult offspring from the same litters consumed a normal or a palm oil-enriched diet for 4 weeks between 140 and 169 days of age. ATB treatment transiently modified maternal fecal microbiota although the minor differences observed for offspring colonic microbiota were nonsignificant. In the short-term, consistently higher HSP27 and HSP70 levels and transiently increased horseradish peroxidase permeability in ATB offspring colon were observed. Importantly, long-term consequences included reduced colonic horseradish peroxidase permeability, and increased colonic digesta alkaline phosphatase (AP) and TLR2- and TLR4-stimulant concentrations in rectal digesta in adult ATB offspring. Inducible HSP27 and HSP70 did not change. Interactions between early ATB treatment and later diet were noted for paracellular permeability and concentrations of colonic digesta AP. In conclusion, our data suggest that early ATB-induced changes in bacterial colonization modulate important aspects of colonic physiology in the short- and long-terms. PMID:25689154

Arnal, Marie-Edith; Zhang, Jing; Erridge, Clett; Smidt, Hauke; Lallès, Jean-Paul

2015-01-01

320

Trypanosoma cruzi: modification of alkaline phosphatase activity induced by trypomastigotes in cultured human placental villi Trypanosoma cruzi: alteração da atividade de fosfatase alcalina induzida por tripomastigotas em culturas de vilos placentários humanos  

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Full Text Available Human term placental villi cultured ''in vitro" were maintained with bloodstream forms of Trypanosoma cruzi during various periods of time. Two different concentrations of the parasite were employed. Controls contained no T. cruzi. The alkaline phosphatase activity was determined in placental villi by electron microscopy and its specific activity in the culture medium by biochemical methods. Results showed that the hemoflagellate produces a significant decrease in enzyme activity as shown by both ultracytochemical and specific activity studies and this activity was lower in cultures with high doses of parasites. The above results indicate that the reduction in enzyme activity coincides with the time of penetration and proliferation of T. cruzi in mammalian cells. These changes may represent an interaction between human trophoblast and T. cruzi.Vilos placentários humanos a termo foram mantidos "in vitro" em interação com formas tripomastigotas sangüíneas de Trypanosoma cruzi, durante diversos períodos de tempo. Foram utilizadas concentrações diferentes de parasitas. Os controles não continham T. cruzi. Determinou-se a atividade de fosfatase alcalina em vilos placentários mediante microscopia eletrônica e sua atividade específica no meio de cultura, mediante métodos bioquímicos. Os resultados mostraram que o hemoflagelado produz uma diminuição significante da atividade enzimática tanto pelos estudos ultracitoquímicos como de atividade específica e esta atividade de fosfatase alcalina foi menor em culturas com altas doses de parasitas. Estes resultados são indicadores de que a redução de atividade enzimática coincide com o tempo de penetração e proliferação do T. cruzi nas células. Estas mudanças podem representar uma interação entre o trofoblasto humano e o T. cruzi.

Ricardo E. Fretes

1990-12-01

321

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

Scientific Electronic Library Online (English)

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

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

2013-12-01

322

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

OpenAIRE

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

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

2012-01-01

323

Dietary intake and serum bone related chemistry and their correlations in postmenopausal Iranian women.  

OpenAIRE

OBJECTIVES To determine dietary intake and bone related chemistry of osteoporosis and their correlations in postmenopausal Iranian women. METHODS A cross-sectional study was carried out on 58 healthy Iranian, postmenopausal women from January 2005 until August 2006, at Sina Hospital, Tabriz, Iran. Serum calcium, phosphorus, magnesium, and alkaline phosphatase were measured using auto analyzer and parathyroid hormone (PTH) by immune radio metric assay. Dietary intake was assessed by 3...

Nazila Farrin; Ostadrahimi, Ali R.; Mahboob, Soltan A.; Sousan Kolahi; Mostafa Ghavami

2008-01-01

324

Electrifying Phosphatases  

Science.gov (United States)

Ci-VSP, a recently described protein with sequence similarity to both the voltage-sensing domain of a voltage-gated potassium channel and the phosphatase PTEN, functions as a transmembrane phosphoinositide phosphatase that is regulated by changes in voltage across the plasma membrane. Ci-VSP dephosphorylated phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3] in vitro, exhibited capacitative currents that resembled ion channel gating currents, and, when coexpressed with potassium channels that are regulated by PtdIns(4,5)P2, conferred sensitivity of potassium current amplitude to prolonged changes in membrane potential. How the voltage-sensing (VS) domain of Ci-VSP communicates with the phosphatase domain, and how the VS domain moves its charges across the membrane electric field in the absence of a transmembrane pore domain, remain to be determined.

Richard Horn (Jefferson Medical College; Department of Physiology and Institute of Hyperexcitability REV)

2005-10-25

325

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

OpenAIRE

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

1996-01-01

326

Atividade da fosfatase alcalina no lavado broncoalveolar de equinos de policiamento montado no Estado do Rio de Janeiro / Alkaline phosphatase activity in bronchoalveolar lavage of police horses in Rio de Janeiro State, Brazil  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese A utilidade da determinação das atividades enzimáticas no trato respiratório posterior como ferramenta diagnóstica já foi demonstrada em várias espécies. Nesse contexto, este trabalho teve por objetivo determinar a atividade da Fosfatase Alcalina (FAL) no lavado broncoalveolar (LBA) de equinos da Po [...] lícia Militar do Estado do Rio de Janeiro, comparando animais sadios com portadores assintomáticos de doença inflamatória das vias aéreas (DIVA). Para tal, foram avaliados 28 animais adultos, machos, sem histórico de doença respiratória nos dois meses anteriores ao estudo, com os resultados dos exames físicos e laboratoriais (FAL sanguínea, hematócrito, leucograma, proteína total e fibrinogênio plasmáticos) dentro dos parâmetros fisiológicos. Os equinos foram divididos em dois grupos de acordo com o resultado da citologia broncoalveolar. A determinação da atividade da FAL foi realizada por meio de espectrofotometria a partir de alíquotas do sobrenadante do LBA preservadas em nitrogênio líquido. Para a estimativa do fluido epitelial pulmonar e da atividade da FAL neste, foi realizada a correção da diluição provocada pelo lavado. Os equinos com contagem diferencial de tipos celulares compatível com DIVA apresentaram atividade de FAL no LBA menor, quando comparados aos animais sadios, podendo essa dosagem ser utilizada como complementação do diagnóstico da DIVA. Abstract in english The use of determining the enzymatic activities in the posterior respiratory tract as a diagnostic tool has already been demonstrated in several species. In this context, this paper aims to determine the activity of alkaline phosphatase (ALP) in the bronchoalveolar lavage (BAL) of horses from the Mi [...] litary Police of the State of Rio de Janeiro, comparing healthy animals with asymptomatic carriers of an inflammatory airway disease (IAD). Twenty-eight adult male animals with no history of respiratory diseases in the last two months prior to the study were studied. Physical exam and blood laboratory test results (ALP, hematocrit, leukogram, total protein and plasma fibrinogen) were within physiological parameters. The equines were separated into two groups according to the results of the bronchoalveolar cytology. The determination of ALP was done by spectrophotometry with aliquots of the supernatant of the BAL preserved in liquid nitrogen. To estimate pulmonary epithelial lining fluid and ALP activity, correction of the dilution caused by the lavage was done. The horses with a cell type differential count compatible with IAD presented a lower ALP activity in BAL when compared to healthy animals, therefore this dosage can be used as a complement in the diagnosis of IAD.

Maria Luisa Lorêdo Abreu, Jorge; Vanessa, Viscardi; Katia Moreira, Silva; Juliana Nabuco Pereira, Otaka; Nayro Xavier de, Alencar; Rodolpho de Almeida, Torres Filho; Daniel Augusto Barroso, Lessa.

2014-01-01

327

Correlation between serum ferritin level and liver function tests in thalassemic patients receiving multiple blood transfusions  

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Full Text Available Background: Multiple blood transfusions are the mainstay of thalassemic patients in order to combat the severe anemia. These frequent blood transfusions result in the excessive iron deposition, leading to multiple injuries to a variety of organs in the body. In response to these injuries, the levels of various enzymes are disturbed. The whole phenomena usually involve the interrelation of one parameter with some other. The present study aimed to estimate the levels of serum ferritin and hepatic enzymes and to find out any possible correlation between them in thalassemic patients receiving multiple blood transfusions. Methods: A total number of 90 thalassemic patients of both sexes ranging from 10-15 years, receiving multiple blood transfusions were included in the present study. Blood samples from all the patients were withdrawn and analyzed for the values of serum ferritin, hemoglobin and hepatic enzymes (serum alanine transaminase, serum aspartate transaminase, serum alkaline phosphatase. Pearson correlation coefficient was applied to observe correlation between serum ferritin level and hepatic enzymes. A P value of and #8804;0.05 was considered statistically significant. Results: The overall values of serum ferritin, and hepatic enzymes (serum Alanine Transaminase, serum Aspartate Transaminase, serum Alkaline Phosphatase were remarkably increased than their normal values. However, hemoglobin level was considerably decreased in thalassemic patients. A weak positive insignificant correlation was observed between serum ferritin with hepatic enzymes and hemoglobin in thalassemic patients. Conclusion: Multiple blood transfusions cause iron overload in the body, which in turn, lead to increased serum ferritin levels in thalassemic patients. High levels of hepatic enzymes are somewhat correlated to serum ferritin concentration. However, the exact reason of elevated levels is still unclear. Further detailed studies should be conducted in order to identify the exact mechanism behind this and to search for the promising correlations of various parameters in thalassemic patients receiving multiple blood transfusions. [Int J Res Med Sci 2014; 2(3.000: 988-994

Mahmood Asif

2014-06-01

328

Comparative study of biochemical technique and radioimmunoassay for the measurement of serum prostatic acid phosphatase. Interest in the diagnosis of prostatic cancer  

International Nuclear Information System (INIS)

The radioimmunoassay of prostatic acid phosphatase and the measurement of L-tartrate labil acid phosphatase by biochemical technique are compared in the diagnosis of prostatic cancer. This study concerning in 122 patients bearing prostatic cancers (40), prostatic adenomas (30) and other solid tumors (52) shows that the sensibility of RIA technique is better than the biochemical one. The positive predictive value of PAP-RIA is 93%. However, seeing that the percentage of positivity of RIA in intracapsular stages rarely exceeds 40%, this test does not allow to increase detection power of early stages. The RIA technique, if it is better than biochemical method will not be effective as a sole screening tool for prostatic cancer and its principal application consists in the follow-up of the therapy of prostatic cancer

329

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

International Nuclear Information System (INIS)

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

330

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

Scientific Electronic Library Online (English)

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

R.L., Amorim; V.M.B.D., Moura; G.W., Di Santis; E.P., Bandarra; C., Padovani.

2004-06-01

331

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

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

R.L. Amorim

2004-06-01

332

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

International Nuclear Information System (INIS)

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

333

Gamma radiation effects on liofilized human serum  

International Nuclear Information System (INIS)

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

334

Clinical significance of serum glycochlicacid detection in diagnosis of intrahepatic cholestasis of pregnancy  

International Nuclear Information System (INIS)

Intrahepatic cholestasis of pregnancy (ICP) occurred in the middle and later phase of pregnancy. ICP had considerable effect on the perinatal babies. To further study the effect of serum glycochlicacid in diagnosis of ICP, serum glycochlicacid was measured by radio-immunoassay in normal pregnancy women and ICP pregnant women. The determination of alanine aminotransferase (ALT) and alkaline phosphatase (ALP) were taken as contrast. Serum glycochlicacid is significantly higher (P < 0.01) in ICP pregnant women than in normal pregnant women. The positive rate of serum glycochlicacid was 100%, the positive rate of ALT was 80%, the positive rate of ALP was 40%. Serum glycochlicacid is the most sensitive serologic index in diagnosis of ICP

335

Investigations of serum HPL during pregnancy using two different radioimmunoassays  

International Nuclear Information System (INIS)

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

336

Effect of Corn Oil on Liver Glycogen Content and Blood Glucose-6-phosphatase Dehydrogenase in Toads Treated with DMBA  

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Full Text Available Environmental factors play an important role in the etiology of several types of cancer, this discovery has led to a great deal of interest in the role of diet in cancer etiology. Fed the Egyptian toad with 0.5 ml corn oil and 0.2 mg DMBA toad/3, 3 times/week increased the incidence of liver tumor (22 out of 50 cases in comparison with toads treated with DMBA alone (16 out of 50 cases. On the ultrastructural level, corn oil increased (a the depletion of glycogen, (b accumulation of fat and lysosomes in toad liver tumor. The biochemical data indicated that glucose-6 phosphatase dehydrogenase in the blood, acid and alkaline phosphatase enzymes activities were increased in serum of toads treated with DMBA and corn oil than animals treated with DMBA alone.

N.E. Abdelmeguid

2000-01-01

337

Alterations in selected serum biochemical constituents in equids after induced hepatic disease.  

Science.gov (United States)

Effects of induced cholestasis and hepatocellular necrosis and of fasting on serum biochemical constituents including bile acids, IgA, bilirubin, alkaline phosphatase, gamma-glutamyltransferase (GGT), arginase, and the clearance of sodium sulfobromophthalein were studied in 4 groups of equids. The reference value for serum bile acids, as determined by an enzymatic colorimetric procedure for horses and ponies was 5.94 +/- 2.72 mumol/L, there being no statistical difference for horses and ponies. Sample collection at time of feeding had no effect on serum bile acid concentration. Seemingly, serum bile acids, arginase, and GGT were the most sensitive indicators of cholestasis and/or hepatocellular necrosis and would form an essential minimum effective battery of tests to diagnose and prognose hepatic disease in equids. These tests provided a measure of hepatobiliary transport function (bile acids), cell necrosis (arginase), and cholestasis (GGT and bile acids). PMID:2889412

Hoffmann, W E; Baker, G; Rieser, S; Dorner, J L

1987-09-01

338

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

International Nuclear Information System (INIS)

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

339

The study of some haematological and serum biochemical parameters of juvenile beluga (Huso huso) fed oligofructose.  

Science.gov (United States)

A study was conducted to investigate the effects of dietary oligofructose (1, 2 and 3%) on the blood profiles of beluga (Huso huso) juveniles (18.77 ± 0.76 g) compared to fish fed an un-supplemented diet. After 7 weeks of feeding on the experimental diets, haematological parameters, metabolic products (cholesterol, glucose and total protein) and serum enzymes (lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase) were measured. Compared to the control group (0% oligofructose), dietary oligofructose had no effect on red blood cell counts (RBC), mean corpuscular volume (MCV), mean cellular haemoglobin (MCH) or mean cell haemoglobin concentration (MCHC) (P > 0.05). However, haemoglobin (Hb) concentration, leucocyte (WBC) levels and the proportion of lymphocytes were significantly higher (P > 0.05) in the 2% oligofructose fed fish than in the 3% oligofructose fed fish. Additionally, haematocrit (Hct) values (P = 0.049) and the proportion of lymphocytes (P ? 0.01) were significantly higher in the 2% oligofructose group than in the control group. Although serum glucose and total protein remained unaffected, serum cholesterol was significantly lower in the 2% oligofructose group than in the control and 3% oligofructose group (P < 0.05). The results of the present study showed that oligofructose had no significant effects on serum lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase. These results indicate that fish blood profiles could be affected by prebiotics, which should be taken into account in future studies. PMID:20658187

Hoseinifar, Seyed Hossein; Mirvaghefi, Alireza; Merrifield, Daniel L; Amiri, Bagher Mojazi; Yelghi, Saeed; Bastami, Kazem Darvish

2011-03-01

340

Partial purification and characterization of phosphotyrosyl-protein phosphatase(s) from human erythrocyte cytosol  

International Nuclear Information System (INIS)

Phosphotyrosyl-protein phosphatase activity of human erythrocyte cytosol can be resolved into two fractions by DEAE-cellulose chromatography followed by P-cellulose chromatography. Both 32P-Tyr-phosphatases are able to dephosphorylate 32P-Tyr of poly (Glu-Tyr) 4:1 but no angiotensin II and synthetic peptide Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Arg-Gly, previously phosphorylated on tyrosine residues by rat spleen tyrosine-protein kinase. Both 32P-Tyr-phosphatase activities distinctly differ from either 32P-Ser-casein phosphatase activity or acid and alkaline p-nitrophenylphosphatase activities with regard to catalytic and physico-chemical properties such as substrate specificity, chromatographic behavior, response to various effectors

341

Tartrate-resistant acid phosphatase as a biomarker of bone turnover in dog / Fosfatase ácida resistente ao tartarato como biomarcador do metabolismo ósseo no cão  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Determinaram-se os valores da atividade da fosfatase ácida resistente ao tartarato (FART) e avaliou-se a sua variabilidade biológica. Neste estudo, foram utilizados nove cães adultos e saudáveis de raça Podengo Português para as determinações das atividades da FART, da fosfatase alcalina total, da i [...] soenzima óssea da fosfatase alcalina e da concentração dos minerais séricos - cálcio, fósforo e magnésio. A atividade sérica obtida da FART foi de 2,19±0,56 UI/mL, com uma variação intra-individual de 18,3% e interindividual de 25,6%. Foram observadas correlações significativas ao longo do tempo entre FART e cálcio (r=-0,3431; P Abstract in english Values of serum tartrate-resistant acid phosphatase ( TRAP) activity were obtained in adult dogs and its biological variability was assessed. Nine healthy skeletally mature Portuguese Podengo dogs were used for the determination of TRAP, total and bone alkaline phosphatase serum activities, and also [...] to study their relationship with serum minerals, namely calcium (Ca), phosphorous (P), and magnesium (Mg). The serum TRAP activity was 2.19±0.56IU/mL, with intra-individual variation of 18.3% and inter-individual variation of 25.6%. Significant correlations were observed between serum TRAP activity and Ca (r=-0.3431; P

C.P, Sousa; F, Nery; J.T, Azevedo; C.A, Viegas; M.E, Gomes; I.R, Dias.

2011-02-01

342

Detection of phosphatase activity in aquatic and terrestrial cyanobacterial strains  

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Full Text Available Cyanobacteria, as highly adaptable microorganisms, are characterized by an ability to survive in different environmental conditions, in which a significant role belongs to their enzymes. Phosphatases are enzymes produced by algae in relatively large quantities in response to a low orthophosphate concentration and their activity is significantly correlated with their primary production. The activity of these enzymes was investigated in 11 cyanobacterial strains in order to determine enzyme synthesis depending on taxonomic and ecological group of cyanobacteria. The study was conducted with 4 terrestrial cyanobacterial strains, which belong to Nostoc and Anabaena genera, and 7 filamentous water cyanobacteria of Nostoc, Oscillatoria, Phormidium and Microcystis genera. The obtained results showed that the activity of acid and alkaline phosphatases strongly depended on cyanobacterial strain and the environment from which the strain originated. Higher activity of alkaline phosphatases, ranging from 3.64 to 85.14 ?molpNP/s/dm3, was recorded in terrestrial strains compared to the studied water strains (1.11-5.96 ?molpNP/s/dm3. The activity of acid phosphatases was higher in most tested water strains (1.67-6.28 ?molpNP/s/dm3 compared to the activity of alkaline phosphatases (1.11-5.96 ?molpNP/s/dm3. Comparing enzyme activity of nitrogen fixing and non-nitrogen fixing cyanobacteria, it was found that most nitrogen fixing strains had a higher activity of alkaline phosphatases. The data obtained in this work indicate that activity of phosphatases is a strain specific property. The results further suggest that synthesis and activity of phosphatases depended on eco-physiological characteristics of the examined cyanobacterial strains. This can be of great importance for the further study of enzymes and mechanisms of their activity as a part of cyanobacterial survival strategy in environments with extreme conditions. [Projekat Ministarstva nauke Republike Srbije, br. III 43002

Babi? Olivera B.

2013-01-01

343

Serum Zinc Values in Adult Patients Undergoing Bone Marrow Transplantation  

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

M Hadjibabaie

2009-07-01

344

The synthesis of Phosphate-repressible alkaline phosphatase do not appear to be regulated by ambient pH in the filamentous mould Neurospora crassa A síntese da fosfatase alcalina Pi-repressível não parece ser regulada pelo pH ambiente no fungo filamentoso Neurospora crassa  

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Full Text Available In order to investigate further the adaptive response of moulds to ambient pH, we have measured by ELISA the pho-2-encoded Pi-repressible alkaline phosphatase synthesised by Neurospora crassa. We showed that the 74A and pho-2A strains of this mould secrete similar amounts of the pho-2-encoded enzyme irrespective of ambient pH, when both the preg and pgov genes are not functional, i.e., in strains nuc-2+ growing under Pi-starvation. This suggests that pho-2, which is responsive to Pi starvation via the action of genes nuc-2, preg, pgov and nuc-1, is not a gene responsive to ambient pH and that the differential glycosylation observed for the Pi-repressible alkaline phosphatase retained by the mycelium at pH 5.6 or secreted into the growth medium at pH 8.0 is the genetic response to ambient pH sensing in N. crassa.Para investigar a resposta adaptativa ao pH ambiente em fungos, foram determinados por ELISA os níveis de fosfatase alcalina Pi-repressível expressada pelo gene pho-2 de Neurospora crassa. Foi mostrado que as linhagens 74A e pho-2A deste fungo secretam quantidades semelhantes da fosfatase alcalina Pi-repressível independentemente do pH ambiente, quando ambos os genes preg e pgov não são funcionais, isto é, quando a linhagem nuc-2+ cresce em condições de limitação em fosfato inorgânico (Pi. Isto sugere que o gene pho-2, o qual é regulado pela ação hieráquica dos genes nuc-2, preg, pgov e nuc-1, é reprimido pelo fosfato inorgânico, mas não responde ao pH ambiente, e que a diferença na glicosilação observada para a fosfatase alcalina Pi-repressível (APase retida no micélio em pH 5,6 ou APase secretada no meio de cultura em pH 8,0 é a resposta genética para o monitoramento do pH ambiente em N. crassa.

Sérgio R. Nozawa

2002-01-01

345

The synthesis of Phosphate-repressible alkaline phosphatase do not appear to be regulated by ambient pH in the filamentous mould Neurospora crassa / A síntese da fosfatase alcalina Pi-repressível não parece ser regulada pelo pH ambiente no fungo filamentoso Neurospora crassa  

Scientific Electronic Library Online (English)

Full Text Available Para investigar a resposta adaptativa ao pH ambiente em fungos, foram determinados por ELISA os níveis de fosfatase alcalina Pi-repressível expressada pelo gene pho-2 de Neurospora crassa. Foi mostrado que as linhagens 74A e pho-2A deste fungo secretam quantidades semelhantes da fosfatase alcalina P [...] i-repressível independentemente do pH ambiente, quando ambos os genes preg e pgov não são funcionais, isto é, quando a linhagem nuc-2+ cresce em condições de limitação em fosfato inorgânico (Pi). Isto sugere que o gene pho-2, o qual é regulado pela ação hieráquica dos genes nuc-2, preg, pgov e nuc-1, é reprimido pelo fosfato inorgânico, mas não responde ao pH ambiente, e que a diferença na glicosilação observada para a fosfatase alcalina Pi-repressível (APase) retida no micélio em pH 5,6 ou APase secretada no meio de cultura em pH 8,0 é a resposta genética para o monitoramento do pH ambiente em N. crassa. Abstract in english In order to investigate further the adaptive response of moulds to ambient pH, we have measured by ELISA the pho-2-encoded Pi-repressible alkaline phosphatase synthesised by Neurospora crassa. We showed that the 74A and pho-2A strains of this mould secrete similar amounts of the pho-2-encoded enzyme [...] irrespective of ambient pH, when both the preg and pgov genes are not functional, i.e., in strains nuc-2+ growing under Pi-starvation. This suggests that pho-2, which is responsive to Pi starvation via the action of genes nuc-2, preg, pgov and nuc-1, is not a gene responsive to ambient pH and that the differential glycosylation observed for the Pi-repressible alkaline phosphatase retained by the mycelium at pH 5.6 or secreted into the growth medium at pH 8.0 is the genetic response to ambient pH sensing in N. crassa.

Sérgio R., Nozawa; Geraldo, Thedei Jr.; Luciana S.P., Crott; José E., Barbosa; Antonio, Rossi.

2002-01-01

346

Solid-phase radioimmunoassay for human prostatic acid phosphatase  

International Nuclear Information System (INIS)

A solid-phase technique for radioimmunoassay of human prostatic acid phosphatase (EC 3.1.3.2) is described. Human prostatic acid phosphatase was purified from prostatic fluid. Monospecific antisera to the purified acid phosphatase were produced in rabbits. Disposable polypropylene tubes were coated with antiserum and used for radioimmunoassay with 125I-acid phosphatase. The nonspecific binding was minimized by saturating the binding sites of the tubes with bovine serum albumin. The working range of the technique was 1 to 30 ng of antigen. The solid-phase radioimmunoassay is rapid, sensitive, and efficient. In preliminary clinical trials it was shown that (a) patients with advanced prostatic cancer had elevated prostatic acid phosphatase levels by both enzymatic assay and radioimmunoassay assays, and (b) patients with other cancers were in the normal range for prostatic acid phosphatase

347

Correlation of serum prostate specific antigen levels and Tc-99m mdp bone scintigraphy in newly diagnosed patients with prostrate cancer (abstract)  

International Nuclear Information System (INIS)

The aim of the study was to evaluate the correlation between serum prostate specific antigen (PSA) level and bone scintigraphy in newly diagnosed untreated prostate cancer patients. The probability of a positive bone scan for metastases was analyzed for different threshold values of prostate specific antigen (PSA), acid phosphastase and alkaline phosphates. Fifty four newly diagnosed untreated prostate cancer patients (mean age, 67 years range, 41 to 94) were included in this study. In each case serum PSA, acid phosphatase and alkaline phosphatase measurements were performed followed by whole body Technetium-99m MDP bone scan. The positive predictive value of serum PSA level for bone metastases at the threshold of 10 ng/ml was 70% whereas the same threshold level of PSA gave a negative predictive value of 100%. We used receiver operating characteristics (ROC) analysis to examine the power of predictive value of each serum test, in predicting the results of the bone scan. We also applied regression analysis for the assessment of correlation between the levels of tumor markers and the extent of bone pathology. It was concluded that bone scintigraphy seems to be unnecessary in evaluation of newly diagnosed untreated prostate cancer in patients with no clinical signs of bone pathology and serum PSA levels of equal to or less than 10 ng/ml. (author)

348

Hepatic changes and serum ferritin in pancreatic cancer and other gastrointestinal diseases: the role of cholestasis.  

Science.gov (United States)

Serum ferritin, prealbumin, pseudocholinesterase, alpha-1-antitrypsin and caeruloplasmin were determined in control subjects and patients with pancreatic cancer, chronic pancreatitis or extra-pancreatic disease mainly of gastrointestinal origin, in order to investigate the different hepatic changes which influence serum ferritin in chronic pancreatic and other digestive diseases. Increased circulating ferritin was found in pancreatic cancer and extra-pancreatic disease when compared to controls. Correlations were detected between ferritin and the other proteins investigated and between ferritin and total bilirubin, alkaline phosphatase and alanine aminotransferase. Multiple regression analysis demonstrated that cholestasis accounts for 45% of circulating ferritin, the acute-phase response accounted for 18% and decreased liver function accounted for 11%. We conclude that the increase in serum ferritin in chronic pancreatic and other gastrointestinal diseases largely depends on liver changes, with cholestasis probably playing a primary role. PMID:2024931

Basso, D; Fabris, C; Del Favero, G; Meggiato, T; Panozzo, M P; Vianello, D; Plebani, M; Naccarato, R

1991-01-01

349

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

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

PRL Silva

2007-12-01

350

Serum MDA, Antioxidant Vitamins and Erythrocytic Antioxidant Enzymes in Chronic Alcoholic Liver Disease – A Case Control Study  

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Full Text Available Objectives: The study aims to estimate the changes in the serum levels of lipid peroxidation product malondialdehyde (MDA, non-enzymatic antioxidants: vitamin A, E and C and erythrocyte enzymatic antioxidants: superoxide dismutase (SOD and catalase(CAT in chronic alcoholic liver disease. Background: Alcohol consumption accounts for about 50% of patients death from end stage liver disease in India. The increased free radical and their metabolites decrease the plasma antioxidants status in chronic alcoholic liver disease (CALD. Method: The study comprised of 100 healthy persons as controls and 100 diagnosed patients of chronic alcoholic liver disease as cases. The estimation of serum MDA, vitamin A, E, C and erythrocyte enzymatic antioxidants SOD and CAT, were carried out along with liver function parameters like serum aspartate amino transferase (AST, serum alanine aminotransferase (ALT, serum alkaline phosphatase (AP, serum gamma glutamyl transferase (GGT, serum total protein, serum albumin, prothrombin time (PT and serum bilirubin. Statistical analysis was done using unpaired “t” test. Result: The levels of serum MDA were significantly increased in patients with CALD (P<0.01 while antioxidants were significantly reduced as compared to controls (P<0.01. Conclusion: Increased levels of lipid peroxides and reduced antioxidants suggest that, oxidative stress plays a vital role in pathogenesis of chronic alcoholic liver disease

Sunita Pujar

2011-10-01

351

The relationship between the degree of liver fibrosis and serum markers in patient with hepatic diseases  

International Nuclear Information System (INIS)

To study the relationship between the degree of liver fibrosis and serum markers in patients with hepatic diseases, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and ?-glutamyl transpeptidase (GGT), total bilirubin (TBIL), albumin (ALB), globulin (GLO), platelet (PLT), prothrombin time (PT), procollagen type III (PIIINP), hyaluronic acid (HA), laminin (LN) and collagen type IV in 114 patients with different causes hepatic disease were determined. The liver puncture biopsy was also carried out to determine the stages of liver fibrosis. The results showed that the serum albumin, globulin, platelet, prothrombin time, PIIINP, HA, collagen type IV had significant difference in different stages of liver fibrosis. The serum platelets and albumin levels were negatively correlated with the degree of liver fibrosis. The serum PT and GLO levels were positively correlated with time course of liver fibrosis. The serum PIIINP, HA and collagen type IV levels were positively correlated with degree of liver fibrosis. The serum albumin, globulin, prothrombin time, platelets, PIIINP, HA, collagen type IV were correlated with the progress of the liver fibrosis. The prothrombin time and platelets have directive significance in the diagnosis of liver cirrhosis and also used to judge the stage of liver fibrosis in some extent. The serum PIIINP, HA and collagen type IV levels may better reflect the process of liver fibrosis.er reflect the process of liver fibrosis. (authors)

352

PHOSPHATASE EXPRESSION BY CHLORELLA VULGARIS (CHLOROPHYCEAE) IS MEDIATED BY INTERNAL PHOSPHORUS LEVELS AND EXTERNAL PH  

Science.gov (United States)

Cultures of Chlorella vulgaris were grown in custom photobioreactors in acid (pH 5.5) and alkaline (pH 7.5) media under phosphate replete and starved conditions. Analysis of differential phosphatase expression indicates that cultures of C. vulgaris grown under alkaline conditions derepressibly expr...

353

Alkaline perturbation  

International Nuclear Information System (INIS)

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

354

Effect of Temperature and storage time on Hepatobiliary enzyme activities in Goat serum  

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Full Text Available The present study was designed and conducted to choose an ideal storage condition for goat sera samples meant for the assay of hepatobiliary enzymes such as, alanine aminotransferases(ALT, aspartate aminotransferases (AST, alkaline phosphatase (ALP and gamma glutamyltran-sferases (GGT by storing at room temperature, 4 ºC and -20 ºC up to 14 days. Gamma glutamyltransferase was found to be the most stable enzyme in all the three storage conditions through out the study period. Alanine aminotransferase was stable only up to 8 days at 4 ºC whereas marked stability was noticed at -20 ºC and room temperature as long as 14 days. Aspartate aminotransferase was more stable at -20 ºC up to14 days and 11 days at 4 ºC whereas at room temperature only 2 days. Alkaline phosphatase showed great variation upon storage as compared to other hepatobiliary enzymes and it is suggested that its estimation should be performed in fresh serum samples to get a more accurate result. From these results it is therefore advisable to consider stability of each serum hepatobiliary enzymes for different animals separately before preserving sera samples to get more valid and reliable result. [Vet. World 2010; 3(6.000: 277-279

P.D. Divya and K.K. Jayavardhanan

2010-12-01

355

Direct Determination of Phosphatase Activity from Physiological Substrates in Cells  

Science.gov (United States)

A direct and continuous approach to determine simultaneously protein and phosphate concentrations in cells and kinetics of phosphate release from physiological substrates by cells without any labeling has been developed. Among the enzymes having a phosphatase activity, tissue non-specific alkaline phosphatase (TNAP) performs indispensable, multiple functions in humans. It is expressed in numerous tissues with high levels detected in bones, liver and neurons. It is absolutely required for bone mineralization and also necessary for neurotransmitter synthesis. We provided the proof of concept that infrared spectroscopy is a reliable assay to determine a phosphatase activity in the osteoblasts. For the first time, an overall specific phosphatase activity in cells was determined in a single step by measuring simultaneously protein and substrate concentrations. We found specific activities in osteoblast like cells amounting to 116 ± 13 nmol min-1 mg-1 for PPi, to 56 ± 11 nmol min-1 mg-1 for AMP, to 79 ± 23 nmol min-1 mg-1 for beta-glycerophosphate and to 73 ± 15 nmol min-1 mg-1 for 1-alpha-D glucose phosphate. The assay was also effective to monitor phosphatase activity in primary osteoblasts and in matrix vesicles. The use of levamisole – a TNAP inhibitor- served to demonstrate that a part of the phosphatase activity originated from this enzyme. An IC50 value of 1.16 ± 0.03 mM was obtained for the inhibition of phosphatase activity of levamisole in osteoblast like cells. The infrared assay could be extended to determine any type of phosphatase activity in other cells. It may serve as a metabolomic tool to monitor an overall phosphatase activity including acid phosphatases or other related enzymes. PMID:25785438

Ren, Zhongyuan; Do, Le Duy; Bechkoff, Géraldine; Mebarek, Saida; Keloglu, Nermin; Ahamada, Saandia; Meena, Saurabh; Magne, David; Pikula, Slawomir; Wu, Yuqing; Buchet, René

2015-01-01

356

Serum chemistry concentrations of captive woolly monkeys (Lagothrix lagotricha).  

Science.gov (United States)

Woolly monkeys (Lagothrix sp.) are threatened species and numerous zoos have failed to sustain successful populations. The most common causes of death in captive woolly monkeys are related to pregnancy and hypertension. The objective of this retrospective study was to evaluate serum concentrations of a large number of captive woolly monkeys to establish baseline means and compare these concentrations with their closest related species to determine potential abnormalities. Serum analyses from 30 woolly monkeys housed at two institutions (Apenheul, The Netherlands and The Louisville Zoo, KY, USA) over 12 yr were collected. The statistical model included gender, age group (young, 0-4 yr of age; middle, 5-9 yr; and old, 10+ yr), and zoological institution. All panel result means were similar to previously reported concentrations for howler (Alouatta sp.) and spider monkeys (Ateles sp.) with the possible exception of alanine aminotransferase and gamma-glutamyl-transferase being higher, whereas creatinine and phosphorus were lower. The serum glucose mean of 6.7 mmol/L is above the baseline range for humans and spider monkeys. Alkaline phosphatase (ALP), alanine aminotransferase, and sodium (Na) were higher in females and magnesium (Mg) was higher in males (Pmonkey health risk were noted and discussed. Future studies are needed to determine free-ranging serum concentrations to elucidate parameters that contain aberrant concentrations and decrease health status. PMID:19360617

Ange-van Heugten, Kimberly; Verstegen, Martin; Ferket, Peter R; Stoskopf, Michael; van Heugten, Eric

2008-05-01

357

Hematologic and serum chemistry reference intervals for free-ranging lions (Panthera leo).  

Science.gov (United States)

Hematologic and serum chemistry values are used by veterinarians and wildlife researchers to assess health status and to identify abnormally high or low levels of a particular blood parameter in a target species. For free-ranging lions (Panthera leo) information about these values is scarce. In this study 7 hematologic and 11 serum biochemistry values were evaluated from 485 lions from the Kruger National Park, South Africa. Significant differences between sexes and sub-adult (? 36 months) and adult (>36 months) lions were found for most of the blood parameters and separate reference intervals were made for those values. The obtained reference intervals include the means of the various blood parameter values measured in captive lions, except for alkaline phosphatase in the subadult group. These reference intervals can be utilized for free-ranging lions, and may likely also be used as reference intervals for captive lions. PMID:23415881

Maas, Miriam; Keet, Dewald F; Nielen, Mirjam

2013-08-01

358

Characterization and sequence of PhoC, the principal phosphate-irrepressible acid phosphatase of Morganella morganii.  

Science.gov (United States)

Phosphatase activities were investigated in Morganella morganii, which is one of the few enterobacterial species producing high-level phosphate-irrepressible acid phosphatase activity (HPAP phenotype), and the gene encoding the major phosphate-irrepressible acid phosphatase was cloned, sequenced, and its product characterized. Using p-nitrophenyl phosphate as substrate, Morganella produced a major phosphate-irrepressible acid phosphatase (named PhoC) which is associated with the HPAP phenotype, a minor phosphate-irrepressible acid phosphatase, and a phosphate-repressible alkaline phosphatase. The presence of the PhoC activity prevented induction of alkaline phosphatase when a PhoC-hydrolysable organic phosphate ester, such as glycerol 2-phosphate, was the sole phosphate source. PhoC is a secreted nonspecific acid phosphatase apparently composed of four 25 kDa polypeptide subunits. The enzyme is resistant to EDTA, P(i), fluoride and tartrate. The M. morganii PhoC showed 84.6% amino acid sequence identity to the PhoN nonspecific acid phosphatase of Providencia stuartii, 45.3% to the PhoN nonspecific acid phosphatase of Salmonella typhimurium, and 37.8% to the principal acid phosphatase (PhoC) of Zymomonas mobilis. Comparison of sequence data and of regulation of these enzymes suggested a different phylogeny of members of this gene family within the Enterobacteriaceae. PMID:8081499

Thaller, M C; Berlutti, F; Schippa, S; Lombardi, G; Rossolini, G M

1994-06-01

359

Níveis séricos de enzimas de função hepática em frangos de corte de criação industrial clinicamente saudáveis Serum levels of hepatic enzyme function in clinically healthy broiler chickens  

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Full Text Available The values for the main hepatic enzymes included in the profiles of screen clinical biochemistry, alanine-aminotransferase (ALT, aspartate aminotransferase (AST, alkaline phosphatase (FA, lactate desidrogenase (LDH and gamaglutamiltransferase (GGT, in samples of serum of broiler chickens in industrial system, clinically healthy, starting from the seventh day of life, until the slaughter (42 days in weekly intervals were determined. Significant variations were not observed in the analyses in relation to the age of the birds for none of the appraised enzymes.

A. Borsa

2006-08-01

360

Osteogenic potential of cryopreserved human bone marrow-derived mesenchymal stem cells cultured with autologous serum.  

Science.gov (United States)

Secondary bone grafting in the alveolar cleft is one of the most important therapeutic modalities for patients with cleft lip and palate. However, in children, harvesting a sufficient amount of bone is difficult, and repeated operations are often required because deformation of the alveolar cleft may occur because of the grafted bone absorption and bone growth, which imposes a heavy burden on the patients. The burden may be reduced if the banking of human bone marrow-derived mesenchymal stem cells (MSCs) could be made possible, that is, if cryopreserved autologous MSCs, those that have been harvested from the patient's own bone marrow, could be cultured and expanded with the patient's own serum and can be thawed and cultivated for grafting at a later date. In the current study, a hybrid-type bone substitute was prepared by thawing and cultivating MSCs that have been cryopreserved for more than 3 months. The hybrid-type bone substitute was implanted subcutaneously in nude mice. At 6 and 9 weeks after grafting, the bone graft was removed, and the osteogenic potential of the cells cultured with autologous serum, as determined by alkaline phosphatase activity and alizarin red S staining, was compared with those cultured with fetal bovine serum. There was no significant difference in the osteogenic potential between MSCs cultured with autologous serum and those cultured with fetal bovine serum. The results suggest the possibility of artificial bone grafting using MSCs cultured with autologous serum and the banking of the cells. PMID:18520385

Matsuo, Aoi; Yamazaki, Yasuharu; Takase, Chikara; Aoyagi, Kazuya; Uchinuma, Eiju

2008-05-01

361

21 CFR 862.1020 - Acid phosphatase (total or prostatic) test system.  

Science.gov (United States)

...AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES CLINICAL...test system is a device intended to measure the activity of the acid phosphatase enzyme in plasma and serum. (b)...

2010-04-01

362

Effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine-intervened serum on the proliferation and osteogenic differentiation of bone marrow stromal cells.  

Science.gov (United States)

The present study aimed to investigate the effect of kidney-reinforcing and marrow-beneficial traditional Chinese medicine (TCM)-intervened (KRMBTI)-serum on the proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs) in rats. Rat BMSCs were isolated and cultured in vitro with various concentrations of serum obtained from rats at different time-points following treatment with low, medium and high doses of KRMBT. The alkaline phosphatase (ALP) activity and proliferation of the BMCSs was assessed to determine the optimal serum sampling time-point and serum concentration. Transforming growth factor (TGF)-?1 expression of the BMSCs was detected using enzyme-linked immunosorbent assay (ELISA), and hepcidin mRNA expression in the rat livers was detected using reverse transcription polymerase chain reaction. The proliferation of BMCSs treated with serum obtained l h after dosing was observed to be significantly higher than that for BMCSs treated with serum obtained at the four other time-points (Phepcidin was observed to be expressed at significantly higher levels in the high-dose group than in the control group, and hepcidin expression was significantly higher after 10 weeks compared with that after five weeks. These findings suggest that KRMBTI-serum increases TGF-?1 and hepcidin expression levels, which may be the mechanism underlying the promotion of osteogenic differentiation induced by KRMBTI-serum in BMSCs. PMID:25452801

Zhou, DA-An; Deng, Yue-Ning; Liu, Lei; Li, Jian-Jun

2015-01-01

363

Immunoassay of serum conjugates of cholic acid in cystic fibrosis.  

Science.gov (United States)

Pre- and post-prandial serum conjugates of cholic acid (SCCA) were measured by radioimmunoassay (RIA) in 83 patients with cystic fibrosis (CF), 14 of whom did not have steatorrhoea, and in 25 controls. Of the CF patients with steatorrhoea, 38% had fasting SCCA levels greater than 3 standard deviations above mean fasting control values, whereas no CF patient without steatorrhoea had elevated fasting SCCA levels. Steatorrhoeic patients with palpable livers had higher pre- and post-prandial SCCA levels. Post-prandial SCCA levels failed to discriminate between control and CF groups however. Other serum tests of liver function, including the aspartate amino transferase, alkaline phosphatase, albumin, gamma globulin, and albumin : globulin ratio, failed to correlate with the SCCA. Changes in serum protein constituents correlated strongly with pulmonary dysfunction. The results suggest that elevation of fasting SCCA levels in CF patients is a more sensitive indicator of liver dysfunction than other tests and is a better discriminator than post-prandial SCCA levels between normal and abnormal liver function. The test is recommended for early detection of liver dysfunction in CF patients. PMID:6901734

Davidson, G P; Corey, M; Morad-Hassel, F; Sondheimer, J M; Crozier, D; Forstner, G G

1980-01-01

364

Serum prohepcidin levels are potential prognostic markers in patients with multiple myeloma  

Science.gov (United States)

Prohepcidin is the prohormone of hepcidin. Anemia is one of the main clinical features in patients with multiple myeloma (MM) and hepcidin may be associated with iron homeostasis in these patients. However, the clinical significance of prohepcidin is not fully understood. In this retrospective study, we measured serum prohepcidin levels using an immunoassay technique to study its clinical significance in 39 MM patients. Serum prohepcidin levels in patients with MM were weakly correlated with alkaline phosphatase (ALP) levels (r=0.32, P=0.048), calculated by Spearman’s rank correlation, but not with other clinical data, including hemoglobin, serum iron or ferritin. In addition, patients with severe renal insufficiency [creatinine clearance (CCr) <50 ml/min] had significantly higher prohepcidin levels compared with patients with mild or no renal insufficiency (CCr ?50 ml/min, P=0.047). In contrast, low serum prohepcidin levels less than 110 ng/ml were an independent predictor of poor overall survival [hazard ratio (HR), 5.29; 95% confidence interval (CI), 1.65–17.03] in addition to serum creatinine levels of at least 2 mg/dl (HR, 5.32; CI, 1.10–25.64), serum calcium (HR, 3.53; CI, 1.01–12.33) and ECOG performance status grade 4 (HR, 4.15; CI, 1.32–13.09) in the multivariate analysis using Cox proportional hazards model. In the subset of 31 MM patients with CCr ?50 ml/min, low serum prohepcidin (HR, 5.65; CI, 1.60–19.95) was an indicator of poor prognosis in multivariate analysis. These results indicate that serum prohepcidin levels may be associated with ALP and renal function but not iron homeostasis, in MM patients. In addition, lower serum prohepcidin levels are potential independent indicators of poor overall survival in MM patients regardless of renal function. PMID:23170109

HARAGUCHI, KOUICHI; UTO, HIROFUMI; OHNOU, NOBUHITO; TOKUNAGA, MASAHITO; TOKUNAGA, MAYUMI; UTSUNOMIYA, ATAE; HANADA, SHUICHI; TSUBOUCHI, HIROHITO

2012-01-01

365

Effects of dietary arsenic levels on serum parameters and trace mineral retentions in growing and finishing pigs.  

Science.gov (United States)

This experiment was conducted to investigate the effect of dietary arsenic (As) levels on growth performance, serum biochemistry, and the retention of iron, copper, and zinc in tissues of growing and finishing pigs. Ninety-six crossbred pigs were randomly allotted to four dietary treatments. The corn-soybean basal diets were supplemented with 0, 10, 20, and 30 mg As/kg. Arsenic trioxide was used as the arsenic source. The feeding experiment lasted for 78 d. The results showed that the high arsenic diet decreased average daily gain (ADG) (ppyruvic transaminase (GPT), and alkaline phosphatase (ALP) activities, and decreased (ptransaminase (GOT) activity, albumin, and cholesterol were not affected (p>0.05). Arsenic feeding elevated (pheart, bile, and lymphaden of intestine mesentery. There were increases in iron levels in liver, bile, spleen, thymus, and pancreas in pigs fed the high As diets (pheart, and serum were decreased by the arsenic treatment (ppigs with arsenic treatment, but decreased (ppigs. PMID:17194918

Wang, L; Xu, Z R; Jia, X Y; Han, X Y

2006-11-01

366

The development of determining human prostatic acid phosphatase by radioimmunoassay  

International Nuclear Information System (INIS)

We purified human prostatic acid phosphatase (hPAP) from prostatic tissues by affinity chromatography, DEAE cellulose and gel filtration and also examined physicochemical properties of highly purified PAP. We developed a double-antibody radioimmunoassay for hPAP in serum, with use of antiserum raised in rabbit against highly purified PAP. The antiserum did not cross react with acid phosphatase from platelets and red blood cells. Experimental detail are outlined to assess the reproducibility and reliability of the method under various conditions. The upper limit of the serum PAP levels in the present assay was set at 3.0 ng/ml by 162 determinations of samples. The serum PAP levels of 2 untreated patients with prostatic carcinoma were higher than 3.0 ng/ml and 39 patients with benign prostatic hyperplasia were an average value of 1.9 ng/ml. (author)

367

Isolation and characterization of a neutral phosphatase from wheat seedlings  

International Nuclear Information System (INIS)

A neutral phosphatase was purified to homogeneity from wheat seedlings. The enzyme was a monomeric glycoprotein exhibiting a molecular weight of 35,000, frictional ratio of 1.22, Stokes' radius of 26 A, and sedimentation coefficient of 3.2 S. That the enzyme was a glycoprotein was surmised from its chromatographic property on Concanavalin A-Sepharose column. The phosphatase activity was assayed using either fructose-2,6-bisphosphate or p-nitrophenyl phosphate as substrate. The phosphatase activity was not affected by high concentrations of chelating agents and did not require the addition of Mg+2 or Ca+2 for its activity. Molybdate, orthovanadate, Zn+2, and Hg+2 were all potent inhibitors of the phosphatase activity. The inhibition by Hg+2 was reversed by dithiothreitol. The enzyme activity was stimulated by Mn+2 about 2-fold. On the other hand, 3-phosphoglycerate, fructose-6-P and Pi as well as polyamines inhibited the enzyme activity. The ability of the neutral phosphatase to dephosphorylate protein phosphotyrosine was also investigated. The phosphotyrosyl-substrates, such as [32P] phosphotyrosyl-poly(Glu, Tyr)n, -alkylated bovine serum albumin, -angiotensin-1, and -band 3 of erythrocytes, were all substrates of the phosphatase. On the other hand, the enzyme had no activity toward protein phosphoserine and protein phosphothreonine

368

Sevelamer controls parathyroid hormone-induced bone disease as efficiently as calcium carbonate without increasing serum calcium levels during therapy with active vitamin D sterols.  

Science.gov (United States)

Little is known about the impact of various phosphate binders on the skeletal lesions of secondary hyperparathyroidism (2 degrees HPT). The effects of calcium carbonate (CaCO3) and sevelamer were compared in pediatric peritoneal dialysis patients with bone biopsy-proven 2 degrees HPT. Twenty-nine patients were randomly assigned to CaCO3 (n = 14) or sevelamer (n = 15), concomitant with either intermittent doses of oral calcitriol or doxercalciferol for 8 mo, when bone biopsies were repeated. Serum phosphorus, calcium, parathyroid hormone (PTH), and alkaline phosphatase were measured monthly. The skeletal lesions of 2 degrees HPT improved with both binders, and bone formation rates reached the normal range in approximately 75% of the patients. Overall, serum phosphorus levels were 5.5 +/- 0.1 and 5.6 +/- 0.3 mg/dl (NS) with CaCO3 and sevelamer, respectively. Serum calcium levels and the Ca x P ion product increased with CaCO3; in contrast, values remained unchanged with sevelamer (9.6 +/- 01 versus 8.9 +/- 0.2 mg/dl; P 10.2 mg/dl) occurred more frequently with CaCO3 (P < 0.01). Baseline PTH levels were 980 +/- 112 and 975 +/- 174 pg/ml (NS); these values decreased to 369 +/- 92 (P < 0.01) and 562 +/- 164 pg/ml (P < 0.01) in the CaCO3 and the sevelamer groups, respectively (NS between groups). Serum alkaline phosphatase levels also diminished in both groups (P < 0.01). Thus, treatment with either CaCO3 or sevelamer resulted in equivalent control of the biochemical and skeletal lesions of 2 degrees HPT. Sevelamer, however, maintained serum calcium concentrations closer to the lower end of the normal physiologic range, thereby increasing the safety of treatment with active vitamin D sterols. PMID:15944337

Salusky, Isidro B; Goodman, William G; Sahney, Shobha; Gales, Barbara; Perilloux, Ashley; Wang, He-Jing; Elashoff, Robert M; Jüppner, Harald

2005-08-01

369

A clinical assessment of the relationship between bone scintigraphy and serum biochemical markers in hemodialysis patients  

International Nuclear Information System (INIS)

Renal osteodystrophy is a metabolic bone disease and a common complication of end-stage chronic renal failure and maintenance dialysis treatment. In this study, we examined the correlation between quantifying bone scintigraphy and serum biochemical markers in hemodialysis patients. Bone scintigraphy with technetium-99m-hydroxy-methylene-diphosphonate (99mTc-HMDP) was performed on 28 patients on maintenance hemodialysis. Bone scintigraphy was performed using a standard protocol and was quantified by setting regions of interest (ROIs) over selected regions. The bone-to-soft-tissue ratio (B/ST ratio) at each region was calculated in all patients. The B/ST ratios were then compared with serum biochemical markers. The B/ST ratio for the skull correlated well with serum bone-specific alkaline phosphatase (BAP) (r=0.735, p<0.001), serum deoxypyridinoline (DPD) (r=0.806, p<0.001) and intact parathyroid hormone (intact PTH) (r=0.701, p<0.001). The B/ST ratio for the lumbar spine correlated with intact PTH (r=0.387, p<0.05) but not with serum BAP or serum DPD. The B/ST ratio for the femoral neck correlated with serum DPD (r=0.431, p<0.05) and intact PTH (r=0.449, p<0.05) but not with serum BAP. Our data suggest that quantitative bone scintigraphy is a sensitive and useful method for evaluating bone metabolism in hemodialysis patients. The B/ST ratio for the skull may reflect changes of bone metabolism in hemodialysis patients. (author). (author)

370

Serum enzyme pattern and local enzyme gradients in chronic chagasic patients.  

Science.gov (United States)

Histochemical studies of myocardial biopsies from chronic chagasic patients at different evolutive stages showed a pattern primarily characterized by a marked increment in tissue enzymes such as mono-amine oxidase and lysosomal acid phosphatase. This cellular damage can be reflected by changes in certain serum enzymes associated with myocardial metabolism, specially in the coronary sinus, where the blood metabolized by the heart is drained. However, little is known about the possible changes in blood enzyme activity during chronic Chagas disease. In this investigation, the activity of the following enzymes glutamic-oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), alkaline phosphatase (ALP), acid maltase (AM), lactate dehydrogenase (LDH), alpha-hydroxybutyric dehydrogenase (alpha-HBDH or LDH1) and creatine phosphokinase (CPK) was measured in blood serum of the superior cava vein (SCV), coronary sinus (CS) and pulmonary (PA) and femoral (FA) arteries of 45 chronic chagasic patients, ages between 20 and 55 yr, at different evolutive stages (groups IA, IB, II and III). The results demonstrate that the average activity of the enzymes studied in chagasic patients, except LDH and CPK, are significantly altered (p GPT, ALP, acid maltase and alpha-HBDH in groups IA and IB is an indication of early myocardial damage in chronic chagasic patients without clinical evidence of cardiac disease. In conclusion, it is suggested that the possible evolutive pattern for myocardial damage could be established by the increment in coronary sinus blood of the enzymes GOT, acid maltase and alpha-HBDH. PMID:12658870

Alarcón-Corredor, Oscar Marino; Carrasco-Guerra, Hugo; Ramírez de Fernández, María; León, Wanda

2002-01-01

371

Radioprotective effect of Panax ginseng on the phosphatases and lipid peroxidation level in testes of Swiss albino mice  

Energy Technology Data Exchange (ETDEWEB)

The Panax ginseng has been used as traditional medicine for past several years among oriental people. The present investigation has been made to assess the radioprotective efficacy of ginseng root extract in the testicular enzymes of Swiss albino mice. The Swiss albino mice were divided into different groups. Ginseng treated group: The animals were administered 10 mg/kg body weight ginseng root extract intraperitoneal (i.p.). Radiation treated group: The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 cm. Combination group: Animals were administered ginseng extract continuously for 4 d and on 4th day they were irradiated to 8 Gy gamma radiation after 30 min of extract administration. The animals from above groups were autopsied on day 1, 3, 7, 14 and 30. Biochemical estimations of acid and alkaline phosphatases and Lipid peroxidation (LPO) in testes were done. In ginseng treated group acid and alkaline phosphatases activity and LPO level did not show any significant alteration. In irradiated animals there was a significant increase in acid phosphatase activity and LPO level. However, significant decline in alkaline phosphatase activity was observed. The treatment of ginseng before irradiation causes significant decrease in acid phosphatase and LPO level and significant increase in alkaline phosphatase activity. One of the cause of radiation damage is lipid peroxidation. Due to lipid peroxidation, lysosomal membrane permeability alters and thus results in release of hydrolytic enzymes. So, an increase in acid phosphatase was noticed after radiation treatment. The alkaline phosphatase activity is associated with membrane permeability and different stages of spermatogenesis. Due to membrane damage and depletion of germ cells of testes after irradiation the enzyme activity was decreased. Ginseng markedly inhibits lipid peroxidation. It acts in indirect fashion to protect radical processes by inhibition of initiation of free radical processes and thus reduces the radiation damages in testes of Swiss albino mice. (author)

Kumar M.; Sharma M.K.; Saxena P.S.; Kumar A. [Rajasthan Univ., Jaipur (India)

2003-03-01

372

Radioprotective effect of Panax ginseng on the phosphatases and lipid peroxidation level in testes of Swiss albino mice  

International Nuclear Information System (INIS)

The Panax ginseng has been used as traditional medicine for past several years among oriental people. The present investigation has been made to assess the radioprotective efficacy of ginseng root extract in the testicular enzymes of Swiss albino mice. The Swiss albino mice were divided into different groups. Ginseng treated group: The animals were administered 10 mg/kg body weight ginseng root extract intraperitoneal (i.p.). Radiation treated group: The animals were exposed to 8 Gy gamma radiation at the dose rate of 1.69 Gy/min at the distance of 80 cm. Combination group: Animals were administered ginseng extract continuously for 4 d and on 4th day they were irradiated to 8 Gy gamma radiation after 30 min of extract administration. The animals from above groups were autopsied on day 1, 3, 7, 14 and 30. Biochemical estimations of acid and alkaline phosphatases and Lipid peroxidation (LPO) in testes were done. In ginseng treated group acid and alkaline phosphatases activity and LPO level did not show any significant alteration. In irradiated animals there was a significant increase in acid phosphatase activity and LPO level. However, significant decline in alkaline phosphatase activity was observed. The treatment of ginseng before irradiation causes significant decrease in acid phosphatase and LPO level and significant increase in alkaline phosphatase activity. One of the cause of radiation damage is lipid peroxidation. Due to lipid peroxidation, lysosomal membrane pe lipid peroxidation, lysosomal membrane permeability alters and thus results in release of hydrolytic enzymes. So, an increase in acid phosphatase was noticed after radiation treatment. The alkaline phosphatase activity is associated with membrane permeability and different stages of spermatogenesis. Due to membrane damage and depletion of germ cells of testes after irradiation the enzyme activity was decreased. Ginseng markedly inhibits lipid peroxidation. It acts in indirect fashion to protect radical processes by inhibition of initiation of free radical processes and thus reduces the radiation damages in testes of Swiss albino mice. (author)

373

Growth and extracellular phosphatase activity of arbuscular mycorrhizal hyphae as influenced by soil organic matter  

DEFF Research Database (Denmark)

Two experiments were set up to investigate the influence of soil organic matter on growth of arbuscular mycorrhizal (AM) hyphae and concurrent changes in soil inorganic P, organic P and phosphatase activity. A sandy loam soil was kept for 14 months under two regimes (outdoor where surplus precipitation leached through the soil, or indoor at constant moisture) with or without 9% (w/w) chopped wheat straw plus mineral N. Then the soils were partially sterilized and placed in two-compartment pots where mycorrhizal or non-mycorrhizal cucumber plants were grown in one root compartment (RC), and soils differing in organic matter were placed in six parallel hyphal compartments (HC) separated from the RC with a 37 mu m mesh. In the first experiment, using Glomus caledonium, hyphal length densities were measured in the HC after 31 days. Added straw increased hyphal length densities by 34 and 62% for soil kept outdoors and indoors, respectively. In the second experiment, using G. invermaium and only soil kept outdoors,three treatments were included: soil with no added straw with or without a new addition of 0.5% (w/w) of ground clover leaves, and soil with 9% straw plus mineral N. After 41 days hyphal length density was twice as high in soil with added straw compared to the two other treatments. Mycorrhizal colonization resulted in lower activity of acid phosphatase in the HC for two out of three treatments. Alkaline phosphatase activity was only decreased by mycorrhiza in soil without organic matter additions. In soil with added clover alkaline phosphatase activity increased due to the presence of mycorrhizal hyphae. We suggest that mycorrhizas may influence the exudation of acid phosphatase by roots. Hyphae of G. invermaium did apparently not excrete extracellular phosphatases, but their presence may have influenced alkaline phosphatase excreted by other microorganisms, probably through competition for nutrients. Phosphatase activity was not correlated with the concentration of labile organic P in soil extracts.

Joner, E.J.; Jakobsen, I.

1995-01-01

374

Retrospective Study of Serum Sclerostin Measurements in Bed Rest Subjects  

Science.gov (United States)

Animal models and human studies suggest that osteocytes regulate the skeleton s response to mechanical unloading at the cellular level in part by an increase in sclerostin, an inhibitor of the anabolic Wnt pathway. However, few studies have reported changes in serum sclerostin in humans exposed to reduced mechanical loading. Thus, we determined changes in serum sclerostin and bone turnover markers in healthy adult men who participated in a controlled bed rest study. Seven healthy adult men (31 +/- 3 yrs old) underwent 90-day six-degree head down tilt bed rest at the University of Texas Medical Branch in Galveston's Institute for Translational Sciences - Clinical Research Center (ITS-CRC). Serum sclerostin, PTH, serum markers of bone turnover (bone specific alkaline phosphatase, RANKL/OPG, and osteocalcin), urinary calcium and phosphorus excretion, and 24 hour pooled urinary markers of bone resorption (NTX, DPD, PYD) were evaluated pre-bed rest (BL), bed rest day 28 (BR-28), bed rest day 60 (BR-60), and bed rest day 90 (BR-90). In addition, bone mineral density (BMD) was assessed by dual-energy X-ray absorptiometry (DXA) at BL, BR-60, and post bed rest day 5 (BR+5). Data are reported as mean +/- standard deviation. We used repeated measures ANOVA to compare baseline values to BR-28, BR-60, and BR-90. RESULTS Consistent with prior reports, BMD declined significantly (1-2% per month) at weight-bearing skeletal sites (spine, hip, femur neck, and calcaneus). Serum sclerostin levels were elevated above BL at BR-28 (+29% +/- 20%, p = 0.003), BR-60 (+42% +/- 31%, p < 0.001), and BR-90 (22% +/- 21%, p = 0.07). Serum PTH levels were reduced at BR-28 (-17% +/- 16%, p = 0.02), BR-60 (-24% +/- 14%, p = 0.03), and returned to baseline at BR-90 (-21% +/- 21%, p = 0.14). Serum bone turnover markers did not change, however urinary bone resorption markers and calcium were significantly elevated following bed rest (p < 0.01). CONCLUSION We observed an increase of serum sclerostin associated with decreased serum PTH and elevated bone resorption markers in otherwise healthy men subjected to long-term immobilization.

Spatz, J. M.; Fields, E. E.; Yu, E. W.; Divieti, Pajevic P.; Bouxsein, M. L.; Sibonga, M. L.; Zwart, S. R.; Smith, S. M.

2011-01-01

375

Radioimmunoassay of human prostate-specific acid phosphatase in the diagnosis and follow-up of therapy of prostatic cancer  

International Nuclear Information System (INIS)

The author describes the development of a radioimmunoassay for the determination of serum prostate-specific acid phosphatase and studies its application to the diagnosis and follow-up of therapy of prostatic carcinoma. (Auth./C.F.)

376

Serum total and bone alkaline phosphatase and tartrate-resistant acid phosphatase activities for the assessment of bone fracture healing in dogs Fosfatase alcalina total, isoenzima óssea da fosfatase alcalina e fosfatase ácida resistente ao tartarato na monitorização da cicatrização de fraturas ósseas  

OpenAIRE

O objetivo deste trabalho foi estudar o padrão de variação da atividade sérica da fosfatase alcalina total (tALP), da isoenzima óssea da fosfatase alcalina (BALP) e da fosfatase ácida resistente ao tartarato (TRAP), assim como a variação da concentração dos minerais séricos durante o processo de cicatrização de fraturas ósseas no cão. A variação sérica destes marcadores do metabolismo ósseo foi avaliada em nove cães com fraturas diafisárias fechadas de ossos longos, subme...

Sousa, C.; Abreu, H.; Viegas, C.; Azevedo, J.; Reis, R.; Gomes, M.; Dias, I.

2011-01-01

377

Serum total and bone alkaline phosphatase and tartrate-resistant acid phosphatase activities for the assessment of bone fracture healing in dogs / Fosfatase alcalina total, isoenzima óssea da fosfatase alcalina e fosfatase ácida resistente ao tartarato na monitorização da cicatrização de fraturas ósseas  

Scientific Electronic Library Online (English)

Full Text Available O objetivo deste trabalho foi estudar o padrão de variação da atividade sérica da fosfatase alcalina total (tALP), da isoenzima óssea da fosfatase alcalina (BALP) e da fosfatase ácida resistente ao tartarato (TRAP), assim como a variação da concentração dos minerais séricos durante o processo de cic [...] atrização de fraturas ósseas no cão. A variação sérica destes marcadores do metabolismo ósseo foi avaliada em nove cães com fraturas diafisárias fechadas de ossos longos, submetidas a tratamento cirúrgico para osteosíntese. Durante o período pós-operatório, sete animais evoluíram no sentido de uma normal união óssea, sendo que dois deles desenvolveram um processo de não união óssea. Foram observados, relativamente à BALP, valores de actividade sérica mais elevados e com diferença estatística (P Abstract in english [...

C., Sousa; H., Abreu; C., Viegas; J., Azevedo; R., Reis; M., Gomes; I., Dias.

1007-10-01

378

Serum total and bone alkaline phosphatase and tartrate-resistant acid phosphatase activities for the assessment of bone fracture healing in dogs Fosfatase alcalina total, isoenzima óssea da fosfatase alcalina e fosfatase ácida resistente ao tartarato na monitorização da cicatrização de fraturas ósseas  

Directory of Open Access Journals (Sweden)

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

C. Sousa

2011-08-01

379

Association of Maternal Serum C- Reactive Protein Levels with Severity of Preeclampsia  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to investigate C-reactive protein (CRP level in preeclampsia (PE and its association with the severity of the disease. This cross-sectional study included 43 women with mild PE, 43 women with severe PE, and 43 healthy pregnant. They were selected in the third trimester of pregnancy in the Afzalipour Hospital, Kerman, Iran, from March 2006 to March 2007. Mean diastolic pressure and level of proteinuria were used as indicators of the severity of the disease. The results were analyzed by t-test and spearman's rank correlation coefficient. Hemoglobin, aspartate and alanine transaminase, creatinine and urine protein excretion, serum CRP, and alkaline phosphatase were higher in women with PE. There were significant correlations between serum CRP levels and diastolic blood pressure (r = 0.5, P = 0, urinary protein excretion (r = 0.5, P = 0, creatinine (r = 0.2, P = 0.003, spartate transaminase (r = 0.3, P = 0, alanine transaminase (r = 0.2, P = 0.006, and Hemoglobin (r = 0.2, P = 0.001. There were a negative correlation between serum CRP and weight of the new born (r = -0.09, P = 0.01 and gestational age in the time of delivery (r = -0.07, P = 0. We showed higher levels of CRP in women with PE. Elevated serum levels of CRP in PE women are, thus, correlated with severity of disease.

Mirzaie Fatemeh

2009-10-01

380

Hematological and serum biochemical values in pregnant and postpartum females of the squirrel monkey (Saimiri sciureus).  

Science.gov (United States)

The hematological and serum biochemical values of a total of 18 pregnant female squirrel monkeys were determined during the pre- and postpartum period. Pregnancy was determined by abdominal palpation in adult females cohabiting with robust males. The mean body weight of the pregnant females gradually increased toward parturition and dramatically decreased at parturition due to delivery of the infant monkey. The red blood cell count, hematocrit and hemoglobin levels diminished toward parturition and then increased to their normal levels by week 6 after delivery. The Wintrobe constant, MCHC, did not fluctuate, but MCH and MCV values increased in late pregnancy and the early nursing periods. The mean white blood cell count varied between 65.8 and 87.3 (x 10(2)/mm3) during the experimental periods. The mean serum total cholesterol concentration and glutamic-oxaloacetic transaminase activity were lower in the pregnant group during the mid- and late gestation stage than in nonpregnant and nursing females. The mean serum total protein and albumin values were lower in pregnant females than in the controls. Alkaline phosphatase activity increased in late pregnancy and the nursing periods. Since the pregnant females examined gave birth to healthy newborn monkeys and nursed them normally, the hematological and serum biochemical measurements should represent the physiological values for squirrel monkeys during pregnancy and the postpartum period. PMID:8689579

Suzuki, T; Suzuki, N; Shimoda, K; Nagasawa, H

1996-01-01

381

Synergistic radioprotective action of imidazole and serotonin on serum and liver enzymes in rats  

International Nuclear Information System (INIS)

One of the most disadvantages of many chemical radioprotectors is that they do not manifest their radioprotective role except when the dose is administered close to the toxic level. The present study has been carried out in order to investigate the possible attenuation of toxicity through the combined treatment with more than one radioprotector at a much lower concentration levels. Imidazole belonging to the heterocyclic nitrogen compounds and serotonin tabulated as a pharmacologic agent have been investigated. The parameters of study for the evaluation of the radioprotective character are the activity levels of certain serum and liver enzymes related to liver function evaluation. Those are GOT, GPT as well as acid and alkaline phosphatases. The results show that whole body gamma irradiation of rats at 6 Gy; affected marked changes in the activity of blood and liver enzymes could be significantly ameliorated through the administration of mixture of imidazole (25 m kg.) and serotonin (15 mg/kg.)

382

Hematology and serum biochemistry values of Culpeo foxes (Lycalopex culpaeus) from central Chile.  

Science.gov (United States)

Hematology and serum biochemistry values were determined for 31 healthy captive and free-ranging Culpeo foxes (Lycalopex culpaeus) sampled in central Chile between 2008 and 2012. The influences of sex, age, and origin (captive versus free-ranging foxes) on the blood parameters were evaluated. The blood values determined were generally comparable to commonly reported values for other wild canid species and the domestic dog. No differences attributable to sex were observed for any parameter. Juveniles had higher levels of alkaline phosphatase and phosphorus and lower values of mean corpuscular hemoglobin concentration, blood urea nitrogen, total protein, and globulin than adult foxes. Captive and free-ranging animals differed in glucose and albumin values. This is the first study on blood parameters of the Culpeo fox and represents a contribution for clinical evaluations of this carnivore in captivity as well as in the wild. PMID:25314826

Rubio, André V; Hidalgo-Hermoso, Ezequiel; Bonacic, Cristian

2014-09-01

383

Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Reversible phosphorylation events within a polymerisation complex have been proposed to modulate capsular polysaccharide synthesis in Streptococcus pneumoniae. Similar phosphatase and kinase genes are present in the exopolysaccharide (EPS biosynthesis loci of numerous lactic acid bacteria genomes. Results The protein sequence deduced from the wzb gene in Lactobacillus rhamnosus ATCC 9595 reveals four motifs of the polymerase and histidinol phosphatase (PHP superfamily of prokaryotic O-phosphatases. Native and modified His-tag fusion Wzb proteins were purified from Escherichia coli cultures. Extracts showed phosphatase activity towards tyrosine-containing peptides. The purified fusion protein Wzb was active on p-nitrophenyl-phosphate (pNPP, with an optimal activity in presence of bovine serum albumin (BSA 1% at pH 7.3 and a temperature of 75°C. At 50°C, residual activity decreased to 10 %. Copper ions were essential for phosphatase activity, which was significantly increased by addition of cobalt. Mutated fusion Wzb proteins exhibited reduced phosphatase activity on p-nitrophenyl-phosphate. However, one variant (C6S showed close to 20% increase in phosphatase activity. Conclusion These characteristics reveal significant differences with the manganese-dependent CpsB protein tyrosine phosphatase described for Streptococcus pneumoniae as well as with the polysaccharide-related phosphatases of Gram negative bacteria.

Gilbert Christophe

2008-04-01

384

A critical evaluation of a specific radioimmunoassay for prostatic acid phosphatase  

International Nuclear Information System (INIS)

A radioimmunoassay (RIA) method for acid phosphatase detection was compared to a standard enzyme assay using sera from 210 normal volunteers and 285 patients with prostatic disease. Statistical and clinical comparisons were made between defined subgroups. All 55 normal females had RIA detectable serum acid phosphatase, implying that this assay cannot be entirely specific for enzyme of prostatic origin. Urinary catheterization did not affect acid phosphatase levels. In all stages of carcinoma there were more acid phosphatase elevations by the RIA method than enzyme method, but neither assay could differentiate intercapsular cancer from benign prostatic hyperplasia. A small number of patients with biopsy proven negative nodules had marginally elevated values, suggesting an obligation for closer follow-up. The RIA method may be superior for monitoring patients with more advanced malignancy. Additional practical advantages of the RIA include relative simplicity and elimination of the special serum handling required for the enzyme assay

385

FOSFATASA ALCALINA (ALP) Y RUNX2 EN CULTIVOS CELULARES DE OSTEOBLASTOS ESTIMULADOS CON CAMPO ELÉCTRICO / ALKALINE PHOSPHATASE (ALP) AND RUNX2 IN CELL CULTURES STIMULATED OSTEOBLASTS ELECTRIC FIELD / FOSFATASE ALCALINA (ALP) E RUNX2 EM CULTIVOS CELULARES DE OSTEOBLASTOS ESTIMULADOS COM CAMPO ELÉTRICO  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in portuguese O objeto deste estudo foi identificar o estímulo elétrico que deve ser aplicado em cultivos celulares de osteoblastos (Ob) para aumentar a expressão do gene da Fosfatase Alcalina (ALP) e o fator de transcrição Runx2. Foram cultivados Ob da American Type Culture Collection (ATCC) Ref. CRL 11372. Os c [...] ultivos foram estimulados a partir do quinto dia, quando as células apresentaram confluência, até o oitavo dia. O estímulo aplicado a cada grupo experimental foi de 100mV, 200mV, 300mV, 400mV e 500mV respectivamente, cultivou-se um grupo de controle não estimulado com cada grupo experimental. O campo elétrico foi gerado com corrente alternada (CA) e aplicou-se mediante duas placas de alumínio localizadas de forma lateral e paralela aos frascos de cultivo de 25cm2. Os níveis de expressão de mRNA foram medidos com a técnica quantitative reverse transcription polymerase chain reaction (qRT-PCR). Com a aplicação do campo gerado com AC aumentou a expressão do fator de transcrição RunX2 em proporção direta ao aumento da voltagem aplicada. A expressão do gene de ALP foi inversamente proporcional à aplicação do estímulo e identificou-se uma diferença significativa entre a presença e ausência do estímulo, sendo maior na ausência do estímulo. O campo elétrico gerou um sinal que pode aumentar ou diminuir a expressão dos genes que mediam a formação de tecido ósseo. No caso de Runx2, favoreceu a diferenciação de células mesenquimais a Ob com a consequente atividade de remodelação e formação do tecido ósseo. Abstract in spanish El objeto de este estudio fue identificar el estímulo eléctrico que debe aplicarse en cultivos celulares de osteoblastos (Ob) para aumentar la expresión del gen de Fosfatasa Alcalina (ALP) y el factor de transcripción Runx2. Se cultivaron Ob de la American Type Culture Collection (ATCC) Ref. CRL 113 [...] 72. Los cultivos se estimularon del día cinco, cuando las células presentaron confluencia, hasta el día ocho. El estímulo aplicado a cada grupo experimental fue de 100mV, 200mV, 300mV, 400mV y 500mV respectivamente, se cultivó un grupo control no estimulado con cada grupo experimental. El campo eléctrico se generó con corriente alterna (AC) y se aplicó mediante dos placas de aluminio ubicadas de forma lateral y paralela a los frascos de cultivo de 25cm2. Los niveles de expresión de mRNA se midieron con la técnica quantitative reverse transcription polymerase chain reaction (qRT-PCR). Con la aplicación de campo generado con AC aumentó la expresión del factor de transcripción RunX2 en proporción directa al aumento del voltaje aplicado. La expresión del gen de ALP fue inversamente proporcional a la aplicación del estímulo y se identificó una diferencia significativa entre la presencia y ausencia del estímulo, siendo mayor en ausencia del estímulo. El campo eléctrico generó una señal que puede aumentar o disminuir la expresión de los genes que median la formación de tejido óseo. En el caso de Runx2, favoreció la diferenciación de células mesenquimales a Ob con la consecuente actividad de remodelación y formación del tejido óseo. Abstract in english The purpose of this study was to identify the electrical fields to be applied in osteoblast (Ob) cell cultures, in order to increase the expression of Alkaline Phosphatase (ALP) gene and the transcription factor Runx2. Ob cultured where from the American Type Culture Collection (ATCC) Ref CRL 11372. [...] Cell Cultures received stimulation at day five, when they showed a confluent monolayer organization until day eight. The stimulus applied to each experimental group was 100mV, 200mV, 300mV, 400mV and 500mV respectively, a control group was cultured without stimulation. The electric field is generated with altern current (AC) and applied with two aluminum plates located parallel to 25cm2 culture flasks. The mRNA expression levels were measured by reverse transcription quantitative technique polymerase chain re

JORGE ARTURO, REY CUBILLOS; LEONARDO, LAREO; SANDRA, GUTIÉRREZ; MARCELA, GODOY CORREDOR.

2012-12-01

386

Pegvisomant-induced serum insulin-like growth factor-I normalization in patients with acromegaly returns elevated markers of bone turnover to normal  

DEFF Research Database (Denmark)

Active acromegaly is associated with increased biochemical markers of bone turnover. Pegvisomant is a GH receptor antagonist that normalizes serum IGF-I in 97% of patients with active acromegaly. We evaluated the effects of pegvisomant-induced serum IGF-I normalization on biochemical markers of bone and soft tissue turnover, as well as levels of PTH and vitamin D metabolites, in 16 patients (nine males; median age, 52 yr; range, 28-78 yr) with active acromegaly (serum IGF-I at least 30% above upper limit of an age-related reference range). Serum procollagen III amino-terminal propeptide (PIIINP) and type I procollagen amino-terminal propeptide, osteocalcin (OC), bone-related alkaline phosphatase, C-terminal cross-linked telopeptide of type I collagen (CTx), albumin-corrected calcium, intact PTH, 25-hydroxy vitamin D, 1,25-dihydroxy vitamin D [1,25-(OH)(2) vit D], urinary type 1 collagen cross-linked N-telopeptide/creatinine ratio, and urinary calcium (24 h collection) were measured (single-batch analysis) at study entry and after IGF-I normalization, along with sera from 32 age- and sex-matched controls. Compared with controls, PIIINP, OC, and CTx were significantly elevated in patients at baseline. Pegvisomant-induced serum IGF-I normalization (699 +/- 76 to 242 +/- 28 micro g/liter, P <0.001) was associated with a significant decrease in PIIINP, markers of bone formation (type I procollagen amino-terminal propeptide, OC, and bone-related alkaline phosphatase), and resorption (CTx and urinary type 1 collagen cross-linked N-telopeptide/creatinine ratio). 1,25-(OH)(2) vit D decreased and intact PTH increased significantly, but 25-hydroxy vitamin D was unaffected. A significant decline in calculated calcium clearance was observed. The decrease in serum IGF-I correlated positively with the decrease of serum PIIINP (r = 0.7, P <0.01). After normalization of serum IGF-I, there was no statistical difference between patients and controls for any parameters for which control data were available. In conclusion, GH excess is associated with increased bone and soft tissue turnover. Pegvisomant-induced normalization of