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Sample records for serum alkaline phosphatase

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

    DEFF Research Database (Denmark)

    Faerk, J; Peitersen, Birgit; Petersen, S; Michaelsen, K F

    2002-01-01

    BACKGROUND: The bone mineral content of premature infants at term is lower than in mature infants at the same postconceptional age. Serum alkaline phosphatase and serum phosphate are often used as indicators of bone mineralisation. OBJECTIVE: To analyse the association between bone mineral content and serum alkaline phosphatase and serum phosphate. METHODS: Serum alkaline phosphatase and phosphate were measured at weekly intervals during admission in 108 premature infants of gestational age belo...

  2. Associations between Renal Hyperfiltration and Serum Alkaline Phosphatase

    OpenAIRE

    Oh, Se Won; Han, Kum Hyun; Han, Sang Youb

    2015-01-01

    Renal hyperfiltration, which is associated with renal injury, occurs in diabetic or obese individuals. Serum alkaline phosphatase (ALP) level is also elevated in patients with diabetes (DM) or metabolic syndrome (MS), and increased urinary excretion of ALP has been demonstrated in patients who have hyperfiltration and tubular damage. However, little was investigated about the association between hyperfiltration and serum ALP level. A retrospective observational study of the 21,308 adults in t...

  3. Alkaline phosphatase bone isoenzyme and osteocalcin in the serum of hyperthyroid cats.

    Science.gov (United States)

    Archer, F J; Taylor, S M

    1996-01-01

    The effect of hyperthyroidism on serum markers for increased bone metabolism and turnover was evaluated in 36 cats with elevated serum levels of thyroxine and alkaline phosphatase. Serum was analyzed for total and ionized calcium and phosphorous. Alkaline phosphatase isoenzymes were separated by agarose gel electrophoresis and osteocalcin was measured by radioimmunoassay. Values for hyperthyroid cats were compared with those for healthy cats. Alkaline phosphatase bone isoenzyme was markedly increased in all 36 hyperthyroid cats. Osteocalcin was increased in 44% of the cats. There was no correlation among the magnitude of increase in alkaline phosphatase bone isoenzyme, osteocalcin, and serum thyroxine concentrations. Increased serum phosphorus was found in 35% of the cats. Total calcium was within the reference range in all cats, while 50% of the cats had reduced levels of serum ionized calcium. We conclude that hyperthyroid cats do have altered bone metabolism, although it is usually clinically insignificant. PMID:9111692

  4. Serum alkaline phosphatase predicts survival outcomes in patients with skeletal metastatic nasopharyngeal carcinoma

    Scientific Electronic Library Online (English)

    Ying, Jin; Mei-Qin, Yuan; Jun-Qing, Chen; Yi-Ping, Zhang.

    2015-04-01

    Full Text Available OBJECTIVE: Bone metastasis is frequently associated with nasopharyngeal carcinoma. The diagnosis and follow-up of bone metastatic patients usually relies on skeletal X-ray and bone scintigraphy, which are time-consuming and costly. This study aimed to evaluate whether serum alkaline phosphatase off [...] ers clinical value in predicting the clinical response and survival outcome for skeletal metastatic nasopharyngeal carcinoma. METHODS: Serum alkaline phosphatase was measured at baseline and then before each cycle of treatment in 416 nasopharyngeal carcinoma patients with bone metastasis. The correlations between the pre-treatment and post-treatment alkaline phosphatase levels and the treatment efficacy were analyzed using the chi-square test. Survival was analyzed using the Kaplan–Meier method and then compared using the log-rank test. RESULTS: Patients with elevated pre-treatment alkaline phosphatase (>110 IU/L) had significantly worse progression-free survival (P

  5. Distinct alkaline phosphatase in serum of patients with lymphatic leukemia and infectious mononucleosis

    Energy Technology Data Exchange (ETDEWEB)

    Neumann, H.; Moran, E.M.; Russell, R.M.; Rosenberg, I.H.

    1974-10-11

    A distinct alkaline phosphatase (phosphatase N) was demonstrated in the serum of patients with acute lymphatic leukemia, chronic lymphatic leukemia, and infectious mononucleosis. This enzyme closely resembles that extracted from the thymus of mice with lymphoma or lymphatic leukemia, both in its electrophoretic mobility and its substrate specificity. The phosphatase N activity was related to the clinical state of patients with lymphatic leukemia and disappeared with recovery from infectious mononucleosis.

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

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    Savi? Ljiljana

    2008-01-01

    Full Text Available Introduction. Many changes happen during growth and development in an organism as a result of important hormone changes, especially biohumoral ones. These changes make a problem when interpreting biochemical results in pediatric population. The most important changes are intensive calcium and phosphorus metabolic turnover in bone tissue with changes in alkaline phosphatase activity as a result of osteoblast activity. The aim of this study was to follow the serum calcium and phosphorus concentration and alkaline phosphatase activity in children 1-15 years old in different growth and development period and of different sexes and to fortify the influence of growth and development dynamics on biohumoral status in healthy male and female children. Material and methods. We evaluated 117 healthy children of both sexes from 1-15 years of age and divided them into three age groups: 1-5, 6-10 and 11-15 years. We followed the serum calcium and phosphorus concentration and alkaline phosphatase activity in different groups and in different sexes. Results and conclusion. Our investigation found significantly higher values of serum calcium in boys than in girls with no important changes between the age groups and significantly higher values of serum phosphorus in the youngest age group in all children and in different sexes with no important sex differences. Alkaline phosphatase activity followed the growth spurt and was the biggest in 6-10 years group in girls and in 11-15 years group in boys.

  7. Pre-transplantation and post-transplantation serum bone alkaline phosphatase levels in renal transplant patients

    OpenAIRE

    Alper GÜMÜ?; Öztürk, Sava?; DÜZ, Muhammed Emin; SARI, Soner; KOLDA?, Macit; AKAYDIN, Mükerrem

    2014-01-01

    Bone Alkaline Phosphatase (BAP) reflects the biosynthetic activity of the osteoblasts. End-stage renal disease and renal transplantation have effects on bone metabolism. The aim of this study was to evaluate serum BAP levels in pre-transplantation (pre-tx) and post-transplantation (post-tx) samples obtained from patients who underwent a renal transplantation. Forty end-stage renal disease patients (16 male and 24 female) undergoing transplantation and 40 healthy individuals (20 female and 20 ...

  8. 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)

  9. ALP (Alkaline Phosphatase) Test

    Science.gov (United States)

    ... Formal name: Alkaline Phosphatase Related tests: AST ; ALT ; GGT ; Bilirubin ; Liver Panel ; Bone Markers ; Alkaline Phosphatase Isoenzymes; ... may be done to determine the cause. A GGT test and/or a test for 5'-nucleotidase ...

  10. 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

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

    OpenAIRE

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

    2011-01-01

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

  12. Association between Serum Alkaline Phosphatase Level and Cerebral Small Vessel Disease

    Science.gov (United States)

    Lee, Han-Bin; Kim, Jinkwon; Kim, Sang-Heum; Kim, Soonhag; Kim, Ok-Joon; Oh, Seung-Hun

    2015-01-01

    Background Serum alkaline phosphatase (ALP) is a marker of vascular calcification. A high serum ALP level is associated with an increase in cardiovascular events, and predicts poor functional outcome in patients with stroke. We investigated whether serum ALP was associated with cerebral small vessel disease (cSVD) and large cerebral artery stenosis (LCAS). Methods We evaluated vascular risk factors, brain magnetic resonance images (MRIs), and MR angiograms from 1,011 neurologically healthy participants. The presence of silent lacunar infarction (SLI) and moderate-to-severe cerebral white matter hyperintensities (MS-cWMH) were evaluated as indices of cSVD on brain MRIs. Findings of extracranial arterial stenosis (ECAS) or intracranial arterial stenosis (ICAS) were considered to be indices of LCAS on MR angiograms. Results Subjects with SLI (odds ratio [OR]: 2.09; 95% confidence interval [CI]: 1.27–3.42; p = 0.004) and MS-cWMH (OR: 1.48; 95% CI; 1.03–2.13, p = 0.036) were significantly more likely to have ALP levels in the third tertile (ALP ? 195 IU/L) than the first tertile (ALP ? 155 IU/L), after adjusting for cardiovascular risk factors. The mean serum ALP level was significantly higher in patients with SLI or MS-cWMH compared to patients without those findings. After adjustment for confounding factors, the multivariate model found that the statistical significance of serum ALP remained when the presence of SLI (OR: 1.05 per 10 IU/L increase in ALP; 95% CI: 1.02–1.08; p = 0.003) or MS-cWMH (OR: 1.03 per 10 IU/L increase in ALP; 95% CI: 1.00–1.06; p = 0.025) were added to the model. There were no differences in the proportions of patients with LCAS, ICAS, and ECAS across the serum ALP tertiles. Conclusions Our study of neurologically healthy participants found a positive association between serum ALP level and indicators of cSVD, but no association between serum ALP level and the indicators of LCAS. PMID:26580067

  13. Serum alkaline phosphatase as a diagnostic criterion in experimental radiation disease

    International Nuclear Information System (INIS)

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

  14. Correlation of Common Biochemical Markers for Bone Turnover, Serum Calcium, and Alkaline Phosphatase in Post-Menopausal Women

    OpenAIRE

    BHATTARAI, Tirtha; Bhattacharya, Koushik; Chaudhuri, Prasenjit; Sengupta, Pallav

    2014-01-01

    The quality of life for women after menopause is one of the key health issues today, and osteoporosis is a silently progressing metabolic bone disease widely prevalent in post-menopausal women in India. Rapid bone loss occurs in post-menopausal women due to hormonal factors that lead to an increased risk of fractures. Thus, the present study was undertaken to observe the serum calcium and alkaline phosphatase (ALP) levels in post-menopausal women as these substances are biochemical markers of...

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

    OpenAIRE

    Bashir Mahmood Bhatti, Tanzeela Talat and Rozina Sardar

    2002-01-01

    In order to estimate serum alkaline phosphatase, cholesterol, calcium and phosphorus during pre-laying and laying reproductive conditions, 60 hens of Desi, Fayoumi, Cross (Rhode Island Red X Fayoumi} and Nick Chick strains were maintained for one year. Five random blood samples from each strain were collected and analyzed during both pre egg laying and egg laying physiological conditions. It was observed that alkaline phosphatase activity increased significantly (P

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

    OpenAIRE

    DOGLIOTTI, Luigi; BERRUTI, Alfredo

    1993-01-01

    Assessment of the response of bone metastases to endocrine or chemotherapy is difficult, and true response rate is probably underestimated by UICC criteria. Biochemical markers of osteoblast activity, which is linked with bone healing, could be useful for early detection of treatment response. We studied changes in osteoblast function, assessed by serial serum total alkaline phosphatase (tALP) at 0, 1, 2, 3 months from the start of systemic therapy, in 31 patients bearing bone metastases from...

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

    OpenAIRE

    Aliyu I; Isah H; Afonja O

    2006-01-01

    Background : The objective of this study was to explore the relationship, if any, between theserum heat-stable alkaline phosphatase (HS-ALP) activity and the blood pressure (BP) of patients with pre-eclampsia and eclampsia. Method : The activity of HS-ALP was measured using the 4– nitrophenyl phosphate (4– NPP) method after incubation at a high temperature of 65 0 C for exactly 30 minutes in one hundred normal pregnant women and in another one hundred with pre-eclampsia...

  18. Serum proteins, trace metals and phosphatases in psoriasis

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    Bhatnagar M

    1994-01-01

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

  19. Evolution of alkaline phosphatases in primates.

    OpenAIRE

    Goldstein, D J; Rogers, C.; HARRIS, H.

    1982-01-01

    Alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] in placenta, intestine, liver, kidney, bone, and lung from a variety of primate species has been characterized by quantitative inhibition, thermostability, and immunological studies. Characteristic human placental-type alkaline phosphatase occurs in placentas of great apes (chimpanzee and orangutan) but not in placentas of other primates, including gibbon. It is also present in trace amounts in hu...

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

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    Han Ju

    2012-02-01

    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.

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

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    Abedini F

    2012-07-01

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

  2. 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...

  3. Negative modulation of alkaline phosphatase and creatine kinase by homobrassinolide

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    G. Nirmal Kumar

    2011-04-01

    Full Text Available

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

    Keywords: Alkaline phosphatase; Creatine kinase; Homobrassinolide; Oxysterol; Phosphorylation; Rate constant.

  4. Negative modulation of alkaline phosphatase and creatine kinase by homobrassinolide

    OpenAIRE

    G. Nirmal Kumar; S Lakshmy; Srikumar, K.

    2011-01-01

    Homobrassinolide is a plant hormone implicated in plant growth and development. Its effect on animal metabolism was less known to date. We have investigated its effect on the marker enzymes such as alkaline phosphatase and creatine kinase in selected rat tissues-brain, heart, liver, kidney, skeletal muscle and testis. Homobrassinolide was administered (66 and 330ng/ Kg body weight) intradermally in male albino wistar strain rats and changes in alkaline phosphatase and creatine kinase...

  5. Effects of Fok-I polymorphism in vitamin D receptor gene on serum 25-hydroxyvitamin D, bone-specific alkaline phosphatase and calcaneal quantitative ultrasound parameters in young adults.

    Science.gov (United States)

    Tanabe, Rieko; Kawamura, Yuka; Tsugawa, Naoko; Haraikawa, Mayu; Sogabe, Natsuko; Okano, Toshio; Hosoi, Takayuki; Goseki-Sone, Masae

    2015-01-01

    Several genes have been implicated as genetic determinants of osteoporosis. Vitamin D receptor (VDR) is an intracellular hormone receptor that specifically binds to the biologically active form of vitamin D, 1-alpha, 25- dihydroxyvitamin D3 [1, 25(OH)2D], and mediates its effects. One of the most frequently studied single nucleotide polymorphisms is the restriction fragment length polymorphism (RFLP) Fok-I (rs2228570). The presence of a Fok-I site, designated f, allows protein translation to initiate from the first ATG. An allele lacking the site (ATG>ACG: designated F), initiates from a second ATG site. In the present study, we explored the effect of the VDR Fok-I genotype on associations among serum bone-specific alkaline phosphatase (ALP), 25- hydroxyvitamin D3 [25(OH)D], 1, 25(OH)2D, and the dietary nutrient intake in healthy young Japanese subjects (n=193). Dietary nutrient intakes were calculated based on 3-day food records before the day of blood examinations. Quantitative ultrasound (QUS) parameters at the right calcaneus (heel bone) were measured. The allele frequencies were 0.622 for the F allele and 0.378 for the f allele in all subjects. Grouped by the VDR genotype, a significant positive correlation between the levels of serum bone-specific ALP and 25(OH)D was observed in the FF-type (p=0.005), but not in the ff-type. In addition, there was a significant positive correlation between the level of serum 25(OH)D and osteo-sono assessment index (OSI) in the FF-type (p=0.008), but not in the ff-type. These results suggest that the level of circulating 25(OH)D is an important factor when assessing the VDR Fok-I polymorphism to prevent osteoporosis. PMID:26078251

  6. Sensitive optical detection of alkaline phosphatase activity with quantum dots

    Energy Technology Data Exchange (ETDEWEB)

    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

    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.

  7. Phosphate Binding in the Active Site of Alkaline Phosphatase and the Interactions of 2-Nitrosoacetophenone with Alkaline Phosphatase-Induced Small Structural Changes

    OpenAIRE

    ZHANG, LE; Buchet, René; Azzar, Gérard

    2004-01-01

    To monitor structural changes during the binding of Pi to the active site of mammalian alkaline phosphatase in water medium, reaction-induced infrared spectroscopy was used. The interaction of Pi with alkaline phosphatase was triggered by a photorelease of ATP from the inactive P3-[1-(2-nitrophenyl)]ethyl ester of ATP. After photorelease, ATP was sequentially hydrolyzed by alkaline phosphatase giving rise to adenosine and three Pi. Although a phosphodiesterase activity was detected prior the ...

  8. 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

    Directory of Open Access Journals (Sweden)

    Helder Jacobina Santos

    2006-03-01

    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.

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

    OpenAIRE

    Lake, B. D.

    1988-01-01

    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...

  10. Alkaline phosphatase prevents platelet stimulation by thromboxane-mimetics.

    OpenAIRE

    Hatmi, M; Haye, B; Gavaret, J. M.; B. B. Vargaftig; Jacquemin, C

    1991-01-01

    1. The effects of alkaline phosphatase on platelet aggregation, secretion and thromboxane B2 (TxB2) generation induced by the full dose-range of common platelet agonists were studied in human platelet-rich plasma and washed platelets. 2. Platelet aggregation and adenosine 5'-triphosphate (ATP) secretion induced by threshold and supramaximal concentrations of arachidonate and stable TxA2 and prostaglandin endoperoxide-mimetics (compounds U46619 and EP171) were abolished in the presence of alka...

  11. phoD Alkaline Phosphatase Gene Diversity in Soil.

    Science.gov (United States)

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

    2015-10-01

    Phosphatase enzymes are responsible for much of the recycling of organic phosphorus in soils. The PhoD alkaline phosphatase takes part in this process by hydrolyzing a range of organic phosphoesters. We analyzed the taxonomic and environmental distribution of phoD genes using whole-genome and metagenome databases. phoD alkaline phosphatase was found to be spread across 20 bacterial phyla and was ubiquitous in the environment, with the greatest abundance in soil. To study the great diversity of phoD, we developed a new set of primers which targets phoD genes in soil. The primer set was validated by 454 sequencing of six soils collected from two continents with different climates and soil properties and was compared to previously published primers. Up to 685 different phoD operational taxonomic units were found in each soil, which was 7 times higher than with previously published primers. The new primers amplified sequences belonging to 13 phyla, including 71 families. The most prevalent phoD genes identified in these soils were affiliated with the orders Actinomycetales (13 to 35%), Bacillales (1 to 29%), Gloeobacterales (1 to 18%), Rhizobiales (18 to 27%), and Pseudomonadales (0 to 22%). The primers also amplified phoD genes from additional orders, including Burkholderiales, Caulobacterales, Deinococcales, Planctomycetales, and Xanthomonadales, which represented the major differences in phoD composition between samples, highlighting the singularity of each community. Additionally, the phoD bacterial community structure was strongly related to soil pH, which varied between 4.2 and 6.8. These primers reveal the diversity of phoD in soil and represent a valuable tool for the study of phoD alkaline phosphatase in environmental samples. PMID:26253682

  12. Chromatographic separation of alkaline phosphatase from dental enamel

    DEFF Research Database (Denmark)

    Moe, D; Kirkeby, S; Salling, E

    1989-01-01

    Alkaline phosphatase (AP) was prepared from partly mineralized bovine enamel by extraction in phosphate buffer, centrifugation and various chromatographic techniques. Chromatofocusing showed that the enamel enzyme possessed five isoelectric points at the acid pH level ranging from pH 5.7 to pH 4.4. Three enzyme peaks were eluted using low pressure chromatography with a Bio-gel column. With a HPLC gel filtration column the separation of the enamel extract resulted in only one peak with AP activit...

  13. ALKALINE PHOSPHATASE ACTIVITY AS A MARKER OF DOG SEMEN FREEZABILITY

    Directory of Open Access Journals (Sweden)

    KOSINIAK-KAMYSZ K.

    2007-01-01

    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.

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

    OpenAIRE

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

    1994-01-01

    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...

  15. Histochemical imaging of alkaline phosphatase using a novel fluorescent substrate.

    Science.gov (United States)

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

    2014-01-01

    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

  16. Identification of bone and liver metastases from breast cancer by measurement of plasma alkaline phosphatase isoenzyme activity.

    OpenAIRE

    Mayne, P D; Thakrar, S; Rosalki, S B; Foo, A Y; Parbhoo, S

    1987-01-01

    Plasma alkaline phosphatase isoenzyme activities were determined in patients with breast cancer to diagnose and monitor bone and liver metastases. Bone alkaline phosphatase activity was increased in 21 of 50 patients (42%) with radiologically confirmed bone metastases, while total alkaline phosphatase activity was increased in only 10 of 50 (20%); liver alkaline phosphatase activity was raised in 12 of 25 patients (48%) with liver metastases. All patients with liver metastases had bone metast...

  17. 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.

  18. Cytochrome c forms complexes and is partly reduced at interaction with GPI-anchored alkaline phosphatase.

    Czech Academy of Sciences Publication Activity Database

    Dadák, V.; Janiczek, O.; Vrána, Old?ich

    2002-01-01

    Ro?. 1570, ?. 1 (2002), s. 9-18. ISSN 0304-4165 Institutional research plan: CEZ:AV0Z5004920 Keywords : cytochrome c * aromatic amino acid * alkaline phosphatase Subject RIV: BO - Biophysics Impact factor: 1.845, year: 2002

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

    International Nuclear Information System (INIS)

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

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

    OpenAIRE

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

    1984-01-01

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

  1. Optical Algal Biosensor using Alkaline Phosphatase for Determination of Heavy Metals

    OpenAIRE

    Durrieu, Claude; Tran-Minh, Canh

    2002-01-01

    A biosensor is constructed to detect heavy metals from inhibition of alkaline phosphatase (AP) present on the external membrane of Chlorella vulgaris microalgae. The microalgal cells are immobilized on removable membranes placed in front of the tip of an optical fiber bundle inside a homemade microcell. C. vulgaris was cultivated in the laboratory and its alkaline phosphatase activity is strongly inhibited in the presence of heavy metals. This property has been used for the determination of t...

  2. Rapidly diverging evolution of an atypical alkaline phosphatase (PhoAaty) in marine phytoplankton: insights from dinoflagellate alkaline phosphatases

    Science.gov (United States)

    Lin, Xin; Wang, Lu; Shi, Xinguo; Lin, Senjie

    2015-01-01

    Alkaline phosphatase (AP) is a key enzyme that enables marine phytoplankton to scavenge phosphorus (P) from dissolved organic phosphorus (DOP) when inorganic phosphate is scarce in the ocean. Yet how the AP gene has evolved in phytoplankton, particularly dinoflagellates, is poorly understood. We sequenced full-length AP genes and corresponding complementary DNA (cDNA) from 15 strains (10 species), representing four classes of the core dinoflagellate lineage, Gymnodiniales, Prorocentrales, Suessiales, and Gonyaulacales. Dinoflagellate AP gene sequences exhibited high variability, containing variable introns, pseudogenes, single nucleotide polymorphisms and consequent variations in amino acid sequence, indicative of gene duplication events and consistent with the “birth-and-death” model of gene evolution. Further sequence comparison showed that dinoflagellate APs likely belong to an atypical type AP (PhoAaty), which shares conserved motifs with counterparts in marine bacteria, cyanobacteria, green algae, haptophytes, and stramenopiles. Phylogenetic analysis suggested that PhoAaty probably originated from an ancestral gene in bacteria and evolved divergently in marine phytoplankton. Because variations in AP amino acid sequences may lead to differential subcellular localization and potentially different metal ion requirements, the multiple types of APs in algae may have resulted from selection for diversifying strategies to utilize DOP in the P variable marine environment. PMID:26379645

  3. 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-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

  4. 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)

  5. An alkaline phosphatase transport mechanism in the pathogenesis of Alzheimer's disease and neurodegeneration.

    Science.gov (United States)

    Pike, Adrianne F; Kramer, Nynke I; Blaauboer, Bas J; Seinen, Willem; Brands, Ruud

    2015-01-25

    Systemic inflammation is associated with loss of blood-brain barrier integrity and neuroinflammation that lead to the exacerbation of neurodegenerative diseases. It is also associated specifically with the characteristic amyloid-? and tau pathologies of Alzheimer's disease. We have previously proposed an immunosurveillance mechanism for epithelial barriers involving negative feedback-regulated alkaline phosphatase transcytosis as an acute phase anti-inflammatory response that hangs in the balance between the resolution and the progression of inflammation. We now extend this model to endothelial barriers, particularly the blood-brain barrier, and present a literature-supported mechanistic explanation for Alzheimer's disease pathology with this system at its foundation. In this mechanism, a switch in the role of alkaline phosphatase from its baseline duties to a stopgap anti-inflammatory function results in the loss of alkaline phosphatase from cell membranes into circulation, thereby decreasing blood-brain barrier integrity and functionality. This occurs with impairment of both amyloid-? efflux and tau dephosphorylating activity in the brain as alkaline phosphatase is replenished at the barrier by receptor-mediated transport. We suggest systemic alkaline phosphatase administration as a potential therapy for the resolution of inflammation and the prevention of Alzheimer's disease pathology as well as that of other inflammation-related neurodegenerative diseases. PMID:25500268

  6. A novel hypothesis for an alkaline phosphatase 'rescue' mechanism in the hepatic acute phase immune response.

    Science.gov (United States)

    Pike, Adrianne F; Kramer, Nynke I; Blaauboer, Bas J; Seinen, Willem; Brands, Ruud

    2013-12-01

    The liver isoform of the enzyme alkaline phosphatase (AP) has been used classically as a serum biomarker for hepatic disease states such as hepatitis, steatosis, cirrhosis, drug-induced liver injury, and hepatocellular carcinoma. Recent studies have demonstrated a more general anti-inflammatory role for AP, as it is capable of dephosphorylating potentially deleterious molecules such as nucleotide phosphates, the pathogenic endotoxin lipopolysaccharide (LPS), and the contact clotting pathway activator polyphosphate (polyP), thereby reducing inflammation and coagulopathy systemically. Yet the mechanism underlying the observed increase in liver AP levels in circulation during inflammatory insults is largely unknown. This paper hypothesizes an immunological role for AP in the liver and the potential of this system for damping generalized inflammation along with a wide range of ancillary pathologies. Based on the provided framework, a mechanism is proposed in which AP undergoes transcytosis in hepatocytes from the canalicular membrane to the sinusoidal membrane during inflammation and the enzyme's expression is upregulated as a result. Through a tightly controlled, nucleotide-stimulated negative feedback process, AP is transported in this model as an immune complex with immunoglobulin G by the asialoglycoprotein receptor through the cell and secreted into the serum, likely using the receptor's State 1 pathway. The subsequent dephosphorylation of inflammatory stimuli by AP and uptake of the circulating immune complex by endothelial cells and macrophages may lead to decreased inflammation and coagulopathy while providing an early upstream signal for the induction of a number of anti-inflammatory gene products, including AP itself. PMID:23899605

  7. Phosphorylation by alkaline phosphatase: immobilization and synthetic potential

    OpenAIRE

    Lara Babich; Joana L. V. M. Peralta; Hartog, Aloysius F.; Ron Wever

    2013-01-01

    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 o...

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

    Scientific Electronic Library Online (English)

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

    2011-12-01

    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

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

    OpenAIRE

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

    2013-01-01

    Differentiating intracellular from extracellular enzyme activity is important in soil enzymology, but not easy. Here, we report on an adjusted sonication method for the separation of intracellular from extracellular phosphatase activity in soil. Under optimal sonication conditions [soil:water ratio ?=? 1/8 (w/v) and power density ?=? 15 watt ml-1], the activity of alkaline phosphomonoesterase (phosphatase) in a Haplic Cambisol soil increased with sonication time in two distinct steps. A first...

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

    Directory of Open Access Journals (Sweden)

    V.I. Krupyanko

    2005-01-01

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

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

    OpenAIRE

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

    2011-01-01

    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 t...

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

    Directory of Open Access Journals (Sweden)

    Vijayaprasad K

    2010-09-01

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

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

    Science.gov (United States)

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

    2010-03-01

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

  14. Distribution of Alkaline Phosphatase, Osteopontin, RANK Ligand and Osteoprotegerin in Calcified Human Carotid Atheroma.

    Science.gov (United States)

    Higgins, Catherine L; Isbilir, Salim; Basto, Pamela; Chen, Iou Yih; Vaduganathan, Muthiah; Vaduganathan, Periyanan; Reardon, Michael J; Lawrie, Gerald; Peterson, Leif; Morrisett, Joel D

    2015-10-01

    Ectopic vascular calcification is a significant component of atherosclerotic disease. Osteopontin (OPN), Osteoprotegerin (OPG), Receptor Activator of NF?B Ligand (RANKL), and alkaline phosphatase (ALP) are each thought to play central roles in the calcification or demineralization of atherosclerotic lesions. Abnormalities in the balance of these proteins may lead to perturbations in bone remodeling and arterial calcification. The purpose of this study was to measure the distribution of these proteins in human carotid lesions and to elucidate possible mechanism(s) whereby they control the deposition or depletion of arterial calcification. Thirty-three patients who had undergone carotid endarterectomy (CEA) within the previous 18 months and 11 control patients were enrolled. CEA specimens were analyzed by EBCT for calcification content in terms of Agatston (AGAT) and Volume scores. CEA specimens were then cut into 5 mm segments which were homogenized and extracted. Extracts were analyzed for tissue levels of calcium, phosphorus, ALP, OPN, RANKL, and OPG. Fasting blood samples were analyzed for the same components. In CEA tissue segments, the calcification levels (CHA AGAT) were inversely associated with the levels of OPG (r = -0.432/-0.579, p < 0.05) and positively associated with the levels of RANKL (r = 0.332/0.415, p < 0.05). In turn, the tissue levels of OPG were associated with homologous serum levels of OPG (r = 0.820/0.389, p < 0.001), and the tissue levels of RANKL were associated with the serum levels of homologous RANKL (r = 0.739/0.666, p < 0.0001). This study suggests that serum levels of OPG and RANKL may be useful biomarkers for estimating the degree of calcification in carotid atherosclerotic lesions. PMID:26307009

  15. Fullerol-fluorescein isothiocyanate phosphorescent labeling reagent for the determination of glucose and alkaline phosphatase.

    Science.gov (United States)

    Liu, Jia-Ming; Wang, Hong-Xin; Zhang, Li-Hong; Zheng, Zhi-Yong; Lin, Shao-Qin; Lin, Li-Ping; Wang, Xin-Xing; Lin, Chang-Qing; Liu, Jian-Qin; Huang, Qi-Tong

    2010-09-15

    The active -OH group in fullerol (F-ol) could react with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form F-ol-(FITC)(n), which could emit room temperature phosphorescence (RTP) signal of F-ol and FITC on acetate cellulose membrane (ACM), respectively. Their RTP signals were enhanced by N,N-dimethylaniline (DMA). The labeling reaction between the -NCS group of FITC in DMA-F-ol-(FITC)(n) and the -NH2 group in wheat germ agglutinin (WGA) produced DMA-F-ol-(FITC)(n)-WGA, which could further take affinity adsorption (AA) reaction with bioactive substances (BS), such as glucose and alkaline phosphatase (AP), to produce DMA-F-ol-(FITC)(n)-WGA-BS. Both of these two products could maintain the good RTP characteristics of F-ol and FITC. Based on the facts above, a new phosphorescent labeling reagent, DMA-F-ol-FITC, was developed, and a new affinity adsorption solid substrate room temperature phosphorimetry (AASSRTP) for the determination of BS was established. This method was applied to the determination of BS in human serum and the diagnosis of diseases, with the results agreeing very well with those of enzyme-linked immunosorbent assay (ELISA). The mechanism of DMA-F-ol-(FITC)(n) labeling of WGA and AASSRTP for the determination of BS is discussed. PMID:20507821

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

    Directory of Open Access Journals (Sweden)

    AKBARI M

    2001-01-01

    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.

  17. Effects of Alkaline Phosphatase Activity on Nucleotide Measurements in Aquatic Microbial Communities †

    OpenAIRE

    Karl, D.M.; Craven, D. B.

    1980-01-01

    Alkaline phosphatase (APase) activity was detected in aquatic microbial assemblages from the subtropics to Antarctica. The occurrence of APase in environmental nucleotide extracts was shown to significantly affect the measured concentrations of cellular nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, guanosine triphosphate, uridine triphosphate, and cytidine triphosphate), adenylate energy charge, and guanosine triphosphate/adenosine triphosphate ratios, w...

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

    OpenAIRE

    Berezhetskyy, A.

    2008-01-01

    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 fou...

  19. Immunoelectron microscopic double labeling of alkaline phosphatase and penicillinase with colloidal gold in frozen thin sections of Bacillus licheniformis 749/C.

    OpenAIRE

    Guan, T; Ghosh, A.; Ghosh, B. K.

    1985-01-01

    The subcellular distribution of alkaline phosphatase and penicillinase was determined by double labeling frozen thin sections of Bacillus licheniformis 749/C with colloidal gold-immunoglobulin G (IgG). Antipenicillinase and anti-alkaline phosphatase antibodies were used to prepare complexes with 5- and 15-nm colloidal gold particles, respectively. The character of the labeling of membrane-bound alkaline phosphatase and penicillinase was different: the immunolabels for alkaline phosphatase (15...

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

    Science.gov (United States)

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

    1999-01-01

    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.

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

    OpenAIRE

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

    1991-01-01

    Lysobacter enzymogenes produces an alkaline phosphatase which is secreted into the medium. The gene for the enzyme (phoA) was isolated from a recombinant lambda library. It was identified within a 4.4-kb EcoRI-BamH1 fragment, and its sequence was determined by the chain termination method. The structural gene consists of an open reading frame which encodes a 539-amino-acid protein with a 29-residue signal sequence, followed by a 119-residue propeptide, the 281-residue mature phosphatase, and ...

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

    OpenAIRE

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

    2008-01-01

    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...

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

    CERN Document Server

    Berezhetskyy, A

    2008-01-01

    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

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

    Directory of Open Access Journals (Sweden)

    N. S. Ahmed

    2010-01-01

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

  5. Alkaline phosphatase activity in the subtropical ocean: insights from nutrient, dust and trace metal addition experiments

    OpenAIRE

    Mahaffey, Claire; Reynolds, Sarah; Davis, Clare E.; Lohan, Maeve C.

    2014-01-01

    Phosphorus is an essential nutrient for all life on earth. In the ocean, the most bioavailable form of phosphorus is inorganic phosphate, but in the extensive subtropical gyres, phosphate concentrations can be chronically low and limit primary productivity and nitrogen fixation. In these regions, organisms produce hydrolytic enzymes, such as alkaline phosphatase (AP), that enable them to utilize the more replete dissolved organic phosphorus (DOP) pool to meet their cellular phosphorus demands...

  6. Amino acid sequence of the cold-active alkaline phosphatase from Atlantic cod (Gadus morhua)

    DEFF Research Database (Denmark)

    Asgeirsson, Bjarni; Nielsen, Berit Noesgaard; Højrup, Peter

    2003-01-01

    Atlantic cod is a marine fish that lives at low temperatures of 0-10 degrees C and contains a cold-adapted alkaline phosphatase (AP). Preparations of AP from either the lower part of the intestines or the pyloric caeca area were subjected to proteolytic digestion, mass spectrometry and amino acid sequencing by Edman degradation. The primary structure exhibits greatest similarity to human tissue non-specific AP (80%), and approximately 30% similarity to AP from Escherichia coli. The key residues ...

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

    OpenAIRE

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

    2013-01-01

    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) ...

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

    OpenAIRE

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

    2014-01-01

    A chronoamperometric method for vanadium ion determination, based on the inhibition of the enzyme alkaline phosphatase, is reported. Screen-printed carbon electrodes modified with gold nanoparticles were used as transducers for the immobilization of the enzyme. The enzymatic activity over 4-nitrophenyl phosphate sodium salt is affected by vanadium ions, which results in a decrease in the chronoamperometric current registered. The developed method has a detection limit of 0.39 ± 0.06 ?M, a rep...

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

    DEFF Research Database (Denmark)

    Clausen, Bettina Hjelm; Fenger, Christina; Finsen, B.

    2013-01-01

    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.

  10. Alkaline Phosphatase Protects Lipopolysaccharide-Induced Early Pregnancy Defects in Mice

    OpenAIRE

    Lei, Wei; Ni, Hua; Herington, Jennifer; Reese, Jeff; Paria, Bibhash C.

    2015-01-01

    Excessive cytokine inflammatory response due to chronic or superphysiological level of microbial infection during pregnancy leads to pregnancy complications such as early pregnancy defects/loss and preterm birth. Bacterial toxin lipopolysaccharide (LPS), long recognized as a potent proinflammatory mediator, has been identified as a risk factor for pregnancy complications. Alkaline phosphatase (AP) isozymes have been shown to detoxify LPS by dephosphorylation. In this study, we examined the ro...

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

    OpenAIRE

    Llinas, Paola; Masella, Michel; Stigbrand, Torgny; Ménez, André; Stura, Enrico A.; Le Du, Marie Hélène

    2006-01-01

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

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

    OpenAIRE

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

    2007-01-01

    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...

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

    OpenAIRE

    Praveen Kumar Vemuri et al

    2012-01-01

    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...

  14. Developmental appearance of Gla proteins (osteocalcin) and alkaline phosphatase in tooth germs and bones of the rat.

    Science.gov (United States)

    Bronckers, A L; Gay, S; Finkelman, R D; Butler, W T

    1987-08-01

    The tissue distribution and developmental appearance of alkaline phosphatase and Gla proteins, relative to mineralization were examined. Undecalcified sections of tooth germs, calvaria and alveolar bone of the rat were immunostained for Gla proteins, (indirect immunofluorescence and avidin-biotin-peroxidase-complex) and then stained for alkaline phosphatase activity. In the growth regions of tooth germs, Gla protein was observed in young odontoblasts and in early predentin, at a stage prior to formation of the first mineralized dentin. Similar results were obtained for bone: young osteoblasts as well as osteoid were immunopositive for Gla protein prior to formation of the first mineralized bone. Histochemical staining revealed that differentiating odontoblasts and osteoblasts exhibited alkaline phosphatase activity at stages prior to appearance of Gla proteins and that cells adjacent to odontoblasts and osteoblasts (not directly involved in the formation of predentin/dentin and osteoid/bone) stained for alkaline phosphatase, but not for Gla proteins. We conclude, that in these bone- and dentin-forming cells, alkaline phosphatase activity is expressed before the appearance of Gla proteins, but that both appear before the onset of mineralization. We also conclude that Gla protein is a more specific marker for bone and dentin formation than histochemical alkaline phosphatase activity, since only the cells directly involved in these processes were immunostained for Gla protein. PMID:3333320

  15. Inhibition kinetics of acid and alkaline phosphatases by atrazine and methomyl pesticides.

    Science.gov (United States)

    El-Aswad, Ahmed F; Badawy, Mohamed E I

    2015-01-01

    The main objective of this work was to investigate the kinetic characteristics of acid and alkaline phosphatases isolated from different sources and to study the effects of the herbicide atrazine and insecticide methomyl on the activity and kinetic properties of the enzymes. Acid phosphatase (ACP) was isolated from the tomato plant (Solanum lycopersicum L. var. lycopersicum); alkaline phosphatase (ALP) was isolated from two sources, including mature earthworms (Aporrectodea caliginosa) and larvae of the Egyptian cotton leafworm (Spodoptera littoralis). The specific activities of the enzymes were 33.31, 5.56 and 0.72 mmol substrate hydrolyzed per minute per milligram protein for plant ACP, earthworms ALP and cotton leafworm ALP, respectively. The inhibition kinetics indicated that atrazine and methomyl caused competitive-non-competitive inhibition of the enzymes. The relationships between estimates of K(m) and V(max) calculated from the Michaelis-Menten equation have been explored. The extent of the inhibition was different, as estimated by the values of the inhibition constant Ki that were found to be 3.34 × 10(-3), 1.12 × 10(-2) and 1.07 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively, with methomyl. In the case of atrazine, K(i) were found to be 8.99 × 10(-3), 3.55 × 10(-2) and 1.36 × 10(-2) mM for plant ACP, earthworms ALP and cotton leafworm ALP, respectively. PMID:25996812

  16. The dermatophyte Trichophyton rubrum secretes an EDTA-sensitive alkaline phosphatase on high-phosphate medium

    Directory of Open Access Journals (Sweden)

    Ferreira-Nozawa Monica S.

    2003-01-01

    Full Text Available In this communication, we show that the growth of isolate H6 of the dermatophyte Trichophyton rubrum on non-buffered medium and under saturating phosphate conditions is dependent on the initial growth pH, with an apparent optimum at pH 4.0. In addition, irrespective of the initial growth pH, the pH of the medium alteredduring cultivation reaching values that ranged from 8.3 to 8.9. Furthermore, this isolate synthesized and secreted almost the same levels of an alkaline phosphatase with an apparent optimum pH ranging from 9.0 to 10.0 when grown on both low- and high-phosphate medium. Also, this alkaline phosphatase is activated by Mg2+ and is EDTA-sensitive. On the other hand, the very low levels of the enzyme retained by the mycelium grown on buffered medium at pH 5.0-5.2 suggest that this enzyme is encoded by an alkaline gene, i.e., a gene responsive to ambient pH signaling.

  17. Inorganic Phosphate Transport in Escherichia coli: Involvement of Two Genes Which Play a Role in Alkaline Phosphatase Regulation

    Science.gov (United States)

    Willsky, Gail R.; Bennett, Robert L.; Malamy, Michael H.

    1973-01-01

    Two classes of alkaline phosphatase constitutive mutations which comprise the original phoS locus (genes phoS and phoT) on the Escherichia coli genome have been implicated in the regulation of alkaline phosphatase synthesis. When these mutations were introduced into a strain dependent on a single system, the pst system, for inorganic phosphate (Pi) transport, profound changes in Pi transport were observed. The phoT mutations led to a complete Pi? phenotype in this background, and no activity of the pst system could be detected. The introduction of the phoS mutations changed the specificity of the pst system so that arsenate became growth inhibitory. Changes in the phosphate source led to changes in the levels of constitutive alkaline phosphatase synthesis found in phoS and phoT mutants. When glucose-6-phosphate or l-?-glycerophosphate was supplied as the sole source of phosphate, phoT mutants showed a 3- to 15- fold reduction in constitutive alkaline phosphatase synthesis when compared to the maximal levels found in limiting Pi media. However, these levels were still 100 times greater than the basal level of alkaline phosphatase synthesized in wild-type strains under these conditions. The phoS mutants showed only a two- to threefold reduction when grown with organic phosphate sources. The properties of the phoT mutants selected on the basis of constitutive alkaline phosphatase synthesis were similar in many respects to those of pst mutants selected for resistance to growth inhibition caused by arsenate. It is suggested that the phoS and phoT genes are primarily involved in Pi transport and, as a result of this function, play a role in the regulation of alkaline phosphatase synthesis. PMID:4570598

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

    Science.gov (United States)

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

    2011-01-01

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

  19. The dermatophyte Trichophyton rubrum secretes an EDTA-sensitive alkaline phosphatase on high-phosphate medium

    OpenAIRE

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

    2003-01-01

    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 app...

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

    Directory of Open Access Journals (Sweden)

    N. S. Ahmed

    2010-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Jayachandran Venkatesan

    2010-10-01

    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.

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

    International Nuclear Information System (INIS)

    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

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

    International Nuclear Information System (INIS)

    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+.

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

    Energy Technology Data Exchange (ETDEWEB)

    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

    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+}.

  5. 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)

    Grunwald, Sandra K.; Krueger, Katherine J.

    2008-01-01

    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…

  6. The cyanobacterium Synechococcus sp. strain PCC 7942 contains a second alkaline phosphatase encoded by phoV.

    Science.gov (United States)

    Wagner, K U; Masepohl, B; Pistorius, E K

    1995-12-01

    A gene (phoV) encoding an alkaline phosphatase from Synechococcus sp. strain PCC 7942 was isolated by screening a plasmid gene bank for expression of alkaline phosphatase activity in Escherichia coli JM103. Two independent clones carrying the same alkaline-phosphatase-encoding gene were isolated. One of these clones (pKW1) was further analysed and the nucleotide sequence of a contiguous 3234 bp DNA fragment was determined. Two complete open reading frames (ORF1 and phoV) and an incomplete ORF3 were identified reading in the same direction. The deduced phoV gene product showed 34% identity to the alkaline phosphatase PhoA from Zymomonas mobilis, and the N-terminal part of the putative ORF3 protein exhibited 57% identity to a protein of unknown function from Frankia sp. Insertional inactivation of the Synechococcus PCC 7942 phoV gene failed, indicating an essential role for either the phoV or the ORF3 gene product. PhoV consists of 550 amino acid residues, resulting in a molecular mass of 61.3 kDa. To overexpress the Synechococcus PCC 7942 phoV gene in E. coli, plasmid pKW1 was transformed into a phoA mutant of E. coli (CC118). In E. coli strain CC118(pKW1) PhoV was expressed constitutively with high rates of activity, and was shown to be membrane associated in the periplasmic space. After partial purification of the recombinant PhoV, it was shown that, like other alkaline phosphatases, the Synechococcus PhoV had a broad pH optimum in the alkaline region and a broad substrate specificity for phosphomonoesters, required Zn2+ for activity, and was inhibited by phosphate. In contrast to several other alkaline phosphatases, PhoV was inhibited by Mn2+. Due to the lack of a Synechococcus PCC 7942 phoV mutant strain, the function of PhoV remains uncertain. However, the present results show that Synechococcus PCC 7942 has a second, probably phosphate-irrepressible, alkaline phosphatase (PhoV, 61.3 kDa) in addition to the phosphate-repressible enzyme (PhoA, 145 kDa) already described. PMID:8574398

  7. Fingerprint deposition on nitrocellulose and polyvinylidene difluoride membranes using alkaline phosphatase.

    Science.gov (United States)

    Kurien, Biji T; Danda, Debashish; Scofield, R Hal

    2015-01-01

    Dactyloscopy or fingerprint identification is a vital part of forensic evidence. Identification with fingerprints has been known since the finding of finger impressions on the clay surface of Babylonian legal contracts almost 4,000 years ago. The skin on the fingers and palms appears as grooves and ridges when observed under a microscope. A unique fingerprint is produced by the patterns of these friction skin ridges. Visible fingerprints can be deposited on solid surfaces. Colored inks have been used to deposit fingermarks on documents. Herein, we show that alkaline phosphatase can be used to transfer prints from fingers or palm to nitrocellulose or polyvinylidene difluoride membranes. The prints can be detected by using the nitro blue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate method of detection. PMID:26044031

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

    Science.gov (United States)

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

    2008-01-01

    Under conditions of starvation and disease, the gut barrier becomes impaired, and trophic feeding to prevent gut mucosal atrophy has become a standard treatment of critically ill patients. However, the mechanisms responsible for the beneficial effects of enteral nutrition have remained a mystery. Using in vitro and in vivo models, we demonstrate that the brush–border enzyme, intestinal alkaline phosphatase (IAP), has the ability to detoxify lipopolysaccharide and prevent bacterial invasion across the gut mucosal barrier. IAP expression and function are lost with starvation and maintained by enteral feeding. It is likely that the IAP silencing that occurs during starvation is a key component of the gut mucosal barrier dysfunction seen in critically ill patients. PMID:18292227

  9. Alkaline Phosphatase and CD34 Reaction of Deciduous Teeth Pulp Stem Cells

    Directory of Open Access Journals (Sweden)

    Fatemeh Abedini

    2007-01-01

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

  10. An electrochemical microRNAs biosensor with the signal amplification of alkaline phosphatase and electrochemical-chemical-chemical redox cycling.

    Science.gov (United States)

    Xia, Ning; Zhang, Youjuan; Wei, Xin; Huang, Yaping; Liu, Lin

    2015-06-01

    MicroRNAs (MiRNAs) have been regarded as clinically important biomarkers and drug discovery targets. In this work, we reported a simple and ultrasensitive electrochemical method for miRNAs detection based on single enzyme amplification and electrochemical-chemical-chemical (ECC) redox cycling. Specifically, upon contact with the target miRNAs, the hairpin structure of biotinylated DNA immobilized on gold electrode was destroyed and the biotin group in DNA was forced away from the electrode surface, allowing for the coupling of streptavidin-conjugated alkaline phosphatase (SA-ALP). Then, ascorbic acid (AA, the enzymatic product of ALP) triggered the ECC redox cycling with ferrocene methanol (FcM) and tris(2-carboxyethyl)phosphine (TCEP) as the redox mediator and the chemical reducing reagent, respectively. The method was more sensitive than that with horseradish peroxidase (HRP) or glucose oxidase (GOx) triggered recycling since one ALP molecule captured by one target miRNA molecule promoted the production of thousands of AA. Analytical merits (e.g., detection limit, dynamic range, specificity, regeneration and reproducibility) were evaluated. The feasibility of the method for analysis of miRNA-21 in human serum has also been demonstrated. PMID:26002330

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

    Science.gov (United States)

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

    2015-03-01

    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

  12. Preparation of bromine-77 labelled monoclonal anti-hPLAP [human placental alkaline phosphatase] antibody using chloramine-T

    International Nuclear Information System (INIS)

    A tumor-associated monoclonal antibody, named 7E8 and raised against human placental alkaline phosphatase (hPLAP), is labelled with bromine-77 by means of chloramine-T. The paper describes optimum radiobromination conditions resulting in 34 % radiochemical yield of labelled antibody with more than 90 % immunoreactivity. (author)

  13. Cooperative Electrostatic Interactions Drive Functional Evolution in the Alkaline Phosphatase Superfamily.

    Science.gov (United States)

    Barrozo, Alexandre; Duarte, Fernanda; Bauer, Paul; Carvalho, Alexandra T P; Kamerlin, Shina C L

    2015-07-22

    It is becoming widely accepted that catalytic promiscuity, i.e., the ability of a single enzyme to catalyze the turnover of multiple, chemically distinct substrates, plays a key role in the evolution of new enzyme functions. In this context, the members of the alkaline phosphatase superfamily have been extensively studied as model systems in order to understand the phenomenon of enzyme multifunctionality. In the present work, we model the selectivity of two multiply promiscuous members of this superfamily, namely the phosphonate monoester hydrolases from Burkholderia caryophylli and Rhizobium leguminosarum. We have performed extensive simulations of the enzymatic reaction of both wild-type enzymes and several experimentally characterized mutants. Our computational models are in agreement with key experimental observables, such as the observed activities of the wild-type enzymes, qualitative interpretations of experimental pH-rate profiles, and activity trends among several active site mutants. In all cases the substrates of interest bind to the enzyme in similar conformations, with largely unperturbed transition states from their corresponding analogues in aqueous solution. Examination of transition-state geometries and the contribution of individual residues to the calculated activation barriers suggest that the broad promiscuity of these enzymes arises from cooperative electrostatic interactions in the active site, allowing each enzyme to adapt to the electrostatic needs of different substrates. By comparing the structural and electrostatic features of several alkaline phosphatases, we suggest that this phenomenon is a generalized feature driving selectivity and promiscuity within this superfamily and can be in turn used for artificial enzyme design. PMID:26091851

  14. Use of alkaline phosphatase staining to differentiate canine osteosarcoma from other vimentin-positive tumors.

    Science.gov (United States)

    Barger, A; Graca, R; Bailey, K; Messick, J; de Lorimier, L-P; Fan, T; Hoffmann, W

    2005-03-01

    Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone neoplasia. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, and plasma cell myeloma. The purpose of this study is to determine the sensitivity and specificity of alkaline phosphatase (ALP) staining to differentiate OSA from other tumors that express vimentin by immunocytochemistry or immunohistochemistry. ALP is a hydrolytic enzyme present in multiple tissues including liver, kidney, intestine, placenta, and bone. Hypothetically, neoplasms actively producing bone should be specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 8-10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate toluidine salt-phosphatase substrate. A positive reaction stains the membrane of the cells gray to black. Samples were counterstained with a Romanowsky's stain to determine whether the sample was of representative cellularity. A total of 61 vimentin-positive neoplasms have been evaluated and confirmed histopathologically. Tumors that expressed vimentin and were positive for ALP included 33 OSAs, one multi-lobular tumor of bone, one amelanotic melanoma, and one chondrosarcoma. Tumors that expressed vimentin and were negative for ALP included chondrosarcomas (three of four), multiple fibrosarcomas, and multiple synovial cell sarcomas. The sensitivity is 100%, and the specificity is 89%. In conclusion, ALP appears to be a highly sensitive and fairly specific marker in the diagnosis of OSA. PMID:15753469

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

    Science.gov (United States)

    Chung, Thomas D Y

    2013-01-01

    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

  16. Determination of trace alkaline phosphatase by solid-substrate room-temperature phosphorimetry based on Triticum vulgare lectin labeled with fullerenol.

    Science.gov (United States)

    Liu, Jia-Ming; Gao, Fei; Huang, Hong-Hua; Zeng, Li-Qing; Huang, Xiao-Mei; Zhu, Guo-Hui; Li, Zhi-Ming

    2008-04-01

    Fullerenol (F) shows a strong and stable room-temperature phosphorescence (RTP) signal on the surface of nitrocellulose membrane (NCM) at lambda ex max/ lambda em max =542.0/709.4 nm. When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal. It was also found that quantitative specific affinity-adsorption reaction can be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the DeltaI(p) value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS-RTP) was established, using fullerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot(-1) (corresponding concentration: 2.8x10(-14) g ml(-1), namely 2.8x10(-16) mol l(-1)). This method with high sensitivity, accuracy, and precision has been successfully applied to the determination of the content of ALP in human serum survey and forecast human disease, and the results are tallied with those using alkaline phosphatase kits. The mechanism for the determination of ALP using AA-SS-RTP was also discussed. PMID:18421752

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-03-01

    In order to clarify the structural basis of the halophilic characteristics of an alkaline phosphatase derived from the moderate halophile Halomonas sp. 593 (HaAP), the tertiary structure of HaAP was determined to 2.1 Å resolution by X-ray crystallography. The structural properties of surface negative charge and core hydrophobicity were shown to be intermediate between those characteristic of halophiles and non-halophiles, and may explain the unique functional adaptation to a wide range of salt concentrations. Alkaline phosphatase (AP) from the moderate halophilic bacterium Halomonas sp. 593 (HaAP) catalyzes the hydrolysis of phosphomonoesters over a wide salt-concentration range (1–4 M NaCl). In order to clarify the structural basis of its halophilic characteristics and its wide-range adaptation to salt concentration, the tertiary structure of HaAP was determined by X-ray crystallography to 2.1 Å resolution. The unit cell of HaAP contained one dimer unit corresponding to the biological unit. The monomer structure of HaAP contains a domain comprised of an 11-stranded ?-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{sup ?} 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.

  18. Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli.

    Science.gov (United States)

    Liu, Wei; Zhang, Rubing; Tian, Ning; Xu, Xin; Cao, Yujing; Xian, Mo; Liu, Huizhou

    2015-09-01

    Geraniol is a valuable acyclic monoterpene alcohol and has many applications in the perfume industries, pharmacy and others. It has been hypothesized that phosphatases can convert geranyl diphosphate (GPP) into geraniol. However, whether and which phosphatases can transform GPP to geraniol has remained unanswered up till now. In this paper, the catalysis abilities of 4 different types of phosphatases were studied with GPP as substrate in vitro. They are bifunctional diacylglycerol diphosphate phosphatase (DPP1) and lipid phosphate phosphatase (LPP1) from Saccharomyces cerevisiae, ADP-ribose pyrophosphatase (NudF) and alkaline phosphatase (PhoA) from Escherichia coli. The results show that just PhoA from E. coli can convert GPP into geraniol. Moreover, in order to confirm the ability of PhoA in vivo, the heterologous mevalonate pathway and geranyl diphosphate synthase gene from Abies grandis were co-overexpressed in E. coli with PhoA gene and 5.3 ± 0.2 mg/l geraniol was produced from glucose in flask-culture. Finally, we also evaluated the fed-batch fermentation of this engineered E. coli and a maximum concentration of 99.3 mg/l geraniol was produced while the conversion efficiency of glucose to geranoid (gram to gram) was 0.51%. Our results offer a new option for geraniol biosynthesis and promote the industrial bio-production of geraniol. PMID:26091008

  19. Cloning and overexpression of alkaline phosphatase PhoK from Sphingomonas sp. strain BSAR-1 for bioprecipitation of uranium from alkaline solutions.

    Science.gov (United States)

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

    2008-09-01

    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 BSAR-1. The recombinant strain very rapidly precipitated >90% of input uranium in less than 2 h from alkaline solutions (pH, 9 +/- 0.2) containing 0.5 to 5 mM of uranyl carbonate, compared to BSAR-1, which precipitated uranium in >7 h. In both strains BSAR-1 and EK4, precipitated uranium remained cell bound. The EK4 cells exhibited a much higher loading capacity of 3.8 g U/g dry weight in 7 h in BSAR-1. The data demonstrate the potential utility of genetically engineering PhoK for the bioprecipitation of uranium from alkaline solutions. PMID:18641147

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

    Directory of Open Access Journals (Sweden)

    Praveen Kumar Vemuri et al

    2012-09-01

    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.

  1. Biodistribution and translational pharmacokinetic modeling of a human recombinant alkaline phosphatase.

    Science.gov (United States)

    Peters, Esther; Stevens, Jasper; Arend, Jacques; Guan, Zheng; Raaben, Willem; Laverman, Peter; Elsas, Andrea van; Masereeuw, Rosalinde; Pickkers, Peter

    2015-11-10

    Clinical trials showed renal protective effects of bovine intestinal alkaline phosphatase (AP) in patients with sepsis-associated acute kidney injury (AKI). Subsequently, a human recombinant chimeric AP (recAP) was developed as a pharmaceutically acceptable alternative. Here, we investigated the biodistribution and pharmacokinetics (PK) of recAP and developed a translational population PK model. Biodistribution was studied during LPS-induced AKI in rats. Iodine-125-labeled recAP was primarily taken up by liver, spleen, adrenals, heart, lungs and kidneys followed by the gastro-intestinal tract and thyroid. Tissue distribution was not critically affected by endotoxemia. PK parameters were determined in rats and minipigs during IV bolus injections of recAP, administered once, or once daily during seven consecutive days. Plasma concentrations of recAP increased with increasing dose and disappeared in a biphasic manner. Exposure to recAP, estimated by AUC and Cmax, was similar on days 1 and 7. Subsequently, population approach nonlinear mixed effects modeling was performed with recAP rat and minipig and biAP phase I PK data. Concentration versus time data was accurately described in all species by a two-compartmental model with allometric scaling based on body weight. This model provides a solid foundation for determining the optimal dose and duration of first-in-man recAP studies. PMID:26325308

  2. Guanine-rich DNA-based peroxidase mimetics for colorimetric assays of alkaline phosphatase.

    Science.gov (United States)

    Yang, Jinjin; Zheng, Lin; Wang, Yu; Li, Wei; Zhang, Jinli; Gu, Junjie; Fu, Yan

    2016-03-15

    DNA-based peroxidase mimetics are facilely constructed through Cu(II)-coordination with different oligonucleotides involving G20, C20, A20 and T20, respectively, with high peroxidase mimicking activity as well as high stability against proteins. Peroxidase-like activities of DNA-Cu(II) complexes are greatly associated with the sequence composition of DNA templates, which decrease in the following order: G20>C20>A20>T20. G20-Cu(II) complex ([Cu(2+)]/[base]=0.05) possesses the Km value of 0.257mM toward 3,3',5,5'-tetramethylbenzidine and 102.3mM toward hydrogen peroxide at 25°C. G20-Cu(II) complexes are employed to develop a colorimetric turn-on assay of alkaline phosphatase with high sensitivity and selectivity, on the basis of pyrophosphate-induced inhibition of their intrinsic peroxidase-like activities. The limit of detection is achieved as 0.84U/L with the linear response region of 20-200U/L. Such colorimetric assay system is probably applicable for the quantitative determination of ALP in biological fluids. PMID:26476012

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

    Directory of Open Access Journals (Sweden)

    Zapryanova Dimitrinka

    2013-09-01

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

  4. Alkaline phosphatase variation during carfilzomib treatment is associated with best response in multiple myeloma patients.

    Science.gov (United States)

    Zangari, Maurizio; Aujay, Monette; Zhan, Fenghuang; Hetherington, Kristina L; Berno, Tamara; Vij, Ravi; Jagannath, Sundar; Siegel, David; Keith Stewart, A; Wang, Luhua; Orlowski, Robert Z; Belch, Andrew; Jakubowiak, Andrzej; Somlo, George; Trudel, Suzanne; Bahlis, Nizar; Lonial, Sagar; Singhal, Seema; Kukreti, Vishal; Tricot, Guido

    2011-06-01

    The ubiquitin-proteasome pathway regulates bone formation through osteoblast differentiation. We analyzed variation alkaline phosphatase (ALP) during carfilzomib treatment. Data from 38 patients enrolled in the PX-171-003 and 29 patients in PX-171-004 studies, for patients with relapsed/refractory myeloma, were analyzed. All patients received 20 mg/m(2) of carfilzomib on Days 1, 2, 8, 9, 15, and 16 of a 28-day cycle. Sixty-seven patients from ALP data were evaluable. In PX-171-003, the ORR (>PR) was 18% and the clinical benefit response (CBR; >MR) was 26%, while in PX-171-004, the ORR was 35.5% overall and 57% in bortezomib-naive patients. ALP increment from baseline was statistically different in patients who achieved ? VGPR compared with all others on Days 1 (P = 0.0049) and 8 (P = 0.006) of Cycle 2. In patients achieving a VGPR or better, ALP increased more than 15 units per liter at Cycle 2 Day 1 over baseline. An ALP increase over the same period of time was seen in 26%, 13% and 11% of patients achieving PR, MR, and SD, respectively. This retrospective analysis of patients with relapsed or refractory myeloma treated with single-agent carfilzomib indicates that early elevation in ALP is associated with subsequent myeloma response. PMID:21477075

  5. Resolvin E1-induced intestinal alkaline phosphatase promotes resolution of inflammation through LPS detoxification.

    Science.gov (United States)

    Campbell, Eric L; MacManus, Christopher F; Kominsky, Douglas J; Keely, Simon; Glover, Louise E; Bowers, Brittelle E; Scully, Melanie; Bruyninckx, Walter J; Colgan, Sean P

    2010-08-10

    Resolvin-E1 (RvE1) has been demonstrated to promote inflammatory resolution in numerous disease models. Given the importance of epithelial cells to coordination of mucosal inflammation, we hypothesized that RvE1 elicits an epithelial resolution signature. Initial studies revealed that the RvE1-receptor (ChemR23) is expressed on intestinal epithelial cells (IECs) and that microarray profiling of cells exposed to RvE1 revealed regulation of inflammatory response gene expression. Notably, RvE1 induced intestinal alkaline phosphatase (ALPI) expression and significantly enhanced epithelial ALPI enzyme activity. One role recently attributed to ALPI is the detoxification of bacterial LPS. In our studies, RvE1-exposed epithelia detoxified LPS (assessed by attenuation of NF-kappaB signaling). Furthermore, in epithelial-bacterial interaction assays, we determined that ALPI retarded the growth of Escherichia coli. To define these features in vivo, we used a murine dextran sulfate sodium (DSS) model of colitis. Compared with vehicle controls, administration of RvE1 resulted in significant improvement of disease activity indices (e.g., body weight, colon length) concomitant with increased ALPI expression in the intestinal epithelium. Moreover, inhibition of ALPI activity resulted in increased severity of colitis in DSS-treated animals and partially abrogated the protective influence of RvE1. Together, these data implicate a previously unappreciated role for ALPI in RvE1-mediated inflammatory resolution. PMID:20660763

  6. Passive immunotherapy of mice bearing Ehrlich ascites tumor expressing human, membrane-bound placental alkaline phosphatase.

    Science.gov (United States)

    Barka, T; Henderson, S; van der Noen, H M

    2000-01-01

    The objective of our study was to test if a tumor expressing a transgene coding for a membrane-bound protein is amenable to immunotherapy by antibodies to the same protein. To this end, we have established an Ehrlich ascites tumor (EAT) cell line, EAT-DAP, stably expressing human, membrane-bound placental alkaline phosphatase (PLAP) by infecting EAT cells (EATC) with the retroviral vector DAP and selecting neomycin-resistant cells. EATC and EAT-DAP cells grew at similar rates in vitro, and produced ascites tumor in Swiss-Webster mice with similar efficiency. We have treated mice bearing EAT-DAP ascites tumor with a mouse monoclonal antibody to human PLAP or with a monoclonal antibody to human C proteins of the heterogenous ribonucleoprotein complex (hnRNP). The average survival of mice treated with anti-hnRNP was 16.4 +/- 1.1 days (n = 8). Treatment with anti-PLAP prolonged the survival of mice; in 4 mice average survival was 23.3 +/- 5.7 days. Four animals, however, survived for 60 days when they were killed and had no visible signs of tumor. These data support the notion that passive immunotherapy using antibodies against a membrane protein, expressed in tumor cells transduced by a viral vector coding for that protein, may be effective in controlling tumor growth. PMID:10754465

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

    Science.gov (United States)

    Sakisaka, Yukihiko; Tsuchiya, Masahiro; Nakamura, Takashi; Tamura, Masato; Shimauchi, Hidetoshi; Nemoto, Eiji

    2015-08-01

    Wnt signaling regulates multiple cellular events such as cell proliferation, differentiation, and apoptosis through ?-catenin-dependent canonical and ?-catenin-independent noncanonical pathways. Canonical Wnt/?-catenin signaling can promote the differentiation of dental follicle cells, putative progenitor cells for cementoblasts, osteoblasts, and periodontal ligament cells, toward a cementoblast/osteoblast phenotype during root formation, but little is known about the biological significance of noncanonical Wnt signaling in this process. We identified the expression of Wnt5a, a representative noncanonical Wnt ligand, in tooth root lining cells (i.e. precementoblasts/cementoblasts) and dental follicle cells during mouse tooth root development, as assessed by immunohistochemistry. Silencing expression of the Wnt5a gene in a dental follicle cell line resulted in enhancement of the Wnt3a (a representative canonical Wnt ligand)-mediated increase in alkaline phosphatase (ALP) expression. Conversely, treatment with recombinant Wnt5a inhibited the increase in ALP expression, suggesting that Wnt5a signaling functions as a negative regulator of canonical Wnt-mediated ALP expression of dental follicle cells. Wnt5a did not affect the nuclear translocation of ?-catenin as well as ?-catenin-mediated transcriptional activation of T-cell factor (Tcf) triggered by Wnt3a, suggesting that Wnt5a inhibits the downstream part of the ?-catenin-Tcf pathway. These findings suggest the existence of a feedback mechanism between canonical and noncanonical Wnt signaling during the differentiation of dental follicle cells. PMID:26112214

  8. Steric hindrance regulated supramolecular assembly between ?-cyclodextrin polymer and pyrene for alkaline phosphatase fluorescent sensing.

    Science.gov (United States)

    Song, Chunxia; Yang, Xiaohai; Wang, Kemin; Wang, Qing; Liu, Jianbo; Huang, Jin; Zhou, Maogui; Guo, Xiaochen

    2016-03-01

    We herein report a strategy for sensitive alkaline phosphatase (ALP) fluorescent sensing based on steric hindrance regulated supramolecular assembly between ?-cyclodextrin polymer (poly?-CD) and pyrene. The fluorescence of pyrene was enhanced more than 10 times through supramolecular assembly with poly?-CD. The 5'-phosphorylated dsDNA probe with pyrene attached on the 3'-terminal could be cleaved by ? exonuclease (? exo), yielding pyrene attached on mononucleotides. Pyrene attached on mononucleotides could easily enter the cavity of poly?-CD, resulting in fluorescence enhancement. When ALP was introduced, it could remove 5'-phosphate groups from dsDNA and then prevented the cleavage of dsDNA. Pyrene attached on dsDNA was difficult to enter the cavity of poly?-CD because of steric hindrance, resulting in an inconspicuous fluorescence enhancement. Owing to the excellent fluorescence enhancement during steric hindrance regulated supramolecular assembly, excellent performance of the assay method was achieved for ALP with a detection limit of 0.04UmL(-1). The detection limit was superior or comparable with the reported methods. Besides, this method was simple in design, avoiding double-labeling of probe. PMID:26679620

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

    Directory of Open Access Journals (Sweden)

    I.R. Inyang

    2011-05-01

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

  10. Effects of polyethylene glycol on bovine intestine alkaline phosphatase activity and stability.

    Science.gov (United States)

    Sekiguchi, Satoshi; Yasukawa, Kiyoshi; Inouye, Kuniyo

    2011-01-01

    In this study, we evaluated the effects of polyethylene glycol (PEG) on bovine intestine alkaline phosphatase (BIALP) activity and stability. In the hydrolysis of p-nitrophenylphosphate (pNPP) at pH 9.8 at 20 °C, the k(cat)/K(m) values of BIALP plus 5-15% w/v free PEG with molecular masses of 1, 2, 6, and 20 kDa (PEG1000, PEG2000, PEG6000, and PEG20000 respectively) were 120-140%, 180-300%, 130-170%, and 110-140% respectively of that of BIALP without free PEG (1.8 µM(-1) s(-1)), indicating that activation by PEG2000 was the highest. Unmodified BIALP plus 5% PEG2000 and BIALP pegylated with 2,4-bis(O-methoxypolyethylene glycol)-6-chloro-s-triazine exhibited 1.3-fold higher activity on average than that of BIALP without free PEG under various conditions, including pH 7.0-10.0 and 20-65 °C. The temperatures reducing initial activity by 50% in 30-min incubation of unmodified BIALP plus 5% PEG2000 and pegylated BIALP were 51 and 47 °C respectively, similar to that of BIALP without free PEG (49 °C). These results indicate that the addition of PEG2000 and pegylation increase BIALP activity without affecting its stability, suggesting that they can be used in enzyme immunoassay with BIALP to increase sensitivity and rapidity. PMID:22056430

  11. Probing the Origin of the Compromised Catalysis of E. coli Alkaline Phosphatase in its Promiscuous Sulfatase Reaction

    OpenAIRE

    Catrina, Irina; O'Brien, Patrick J.; Purcell, Jamie; Nikolic-Hughes, Ivana; Zalatan, Jesse G.; Hengge, Alvan C.; Herschlag, Daniel

    2007-01-01

    The catalytic promiscuity of E. coli alkaline phosphatase (AP) and many other enzymes provides a unique opportunity to dissect the origin of enzymatic rate enhancements via a comparative approach. Here we use kinetic isotope effects (KIEs) to explore the origin of the 109-fold greater catalytic proficiency by AP for phosphate monoester hydrolysis relative to sulfate monoester hydrolysis. The primary 18O KIEs for the leaving group oxygen atoms in the AP-catalyzed hydrolysis of p-nitrophenyl ph...

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

    OpenAIRE

    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

    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...

  13. Correlation between Biochemical Parameters and Production Traits in Merino Cross Sheep III-Glucose, Alkaline Phosphatase, Ceruloplasmin

    OpenAIRE

    MERT, Nihat

    2003-01-01

    A group of 111 sheep raised in the Band?rma Sheep Research Institute were used as living materials for this study. Blood samples from 31 Hampshire Down x Merino, 33 Dorset Down x Merino, 27 German Black Headed x Merino and 20 Border Leicester x Merino crosses were taken. Plasma glucose, alkaline phosphatase (ALP) and ceruloplasmin levels were determined. Some production traits such as greasy fleece weight, staple length, fiber diameter, ondulance, lamb birth weight, reproduction traits and th...

  14. HISTOENZYMOLOGICAL DEMONSTRATION OF ALKALINE PHOSPHATASE ACTIVITY IN THE ANTERIOR INTESTINE OF HgCl - TREATED HERBIVOROUS 2 FISH, LABEO ROHITA.

    OpenAIRE

    Dhirender; Jagdish Prasad

    2014-01-01

    Present studies incorporate enterotoxic effects of HgCl on the relative distribution of alkaline 2 phosphatase (AKP) activity in the anterior intestine of fish, Labeo rohita. It was observed that there appears to be slightly decreased AKP activity accompanied by histolytic changes in the epithelial cells lining of intestinal villi and intestinal glands of the anterior intestine of treated fresh water fish. However, in control, these changes were invisible.

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

    OpenAIRE

    Park, Jong Hwa; Park, Chang-Ho; CHUNG, IN SIK

    1997-01-01

    Recombinant alkaline phosphatase expressed in insect cells was concentrated by a factor of one and half times at a separation efficiency of 54.2% using hydrogel ultrafiltration. Enzyme concentration was confirmed by SDS-PAGE as well as by spectrophotometric measurement. Wild and recombinant Autographa californica nuclear polyhedrosis viruses (AcNPV) were concentrated 1.4 and 1.6 times of the feed solution at 48.5 and 60.0% separation efficiency, respectively. Hydrogel ultrafiltration appears ...

  16. Alkaline phosphatase activity as a tool for assessing nutritional conditions in the seagrass Posidonia oceanica.(L.) Delile

    OpenAIRE

    Invers i Brunet, Olga; Pérez Vallmitjana, Marta; Romero, Javier

    1995-01-01

    The effects of experimental phosphorus enrichments on alkaline phosphatase activity (APA) of the seagrass Posidonia oceanica (L.) Delile were tested. Short-term additions (as phosphate, 12 hours, in the laboratory) decreased APA by 18-2896, depending on the plant part considered (roots, young leaves or old leaves). The values of APA after this treatment were well correlated with interna1 phosphorus pools (as P concentration in plant tissues). Long-term additions (as phosphate, added to the se...

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

    OpenAIRE

    Chida, Kohsuke; Taguchi, Meiko

    2011-01-01

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

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

    OpenAIRE

    Sebastián-Serrano, Álvaro; de Diego-García, Laura; Martínez-Frailes, Carlos; Ávila, Jesús; Zimmermann, Herbert; Millán, José Luis; Miras-Portugal, María Teresa; Díaz-Hernández, Miguel

    2014-01-01

    Tissue-nonspecific alkaline phosphatase (TNAP) is one of the four isozymes in humans and mice that have the capacity to hydrolyze phosphate groups from a wide spectrum of physiological substrates. Among these, TNAP degrades substrates implicated in neurotransmission. Transgenic mice lacking TNAP activity display the characteristic skeletal and dental phenotype of infantile hypophosphatasia, as well as spontaneous epileptic seizures and die around 10 days after birth. This physiopathology, lin...

  19. Role of Alkaline Phosphatase from Manduca sexta in the Mechanism of Action of Bacillus thuringiensis Cry1Ab Toxin*

    OpenAIRE

    Arenas, Iván; Bravo, Alejandra; Soberón, Mario; Gómez, Isabel

    2010-01-01

    Cry toxins produced by Bacillus thuringiensis have been recognized as pore-forming toxins whose primary action is to lyse midgut epithelial cells in their target insect. In the case of the Cry1A toxins, a prepore oligomeric intermediate is formed after interaction with cadherin receptor. The Cry1A oligomer then interacts with glycosylphosphatidylinositol-anchored receptors. Two Manduca sexta glycosylphosphatidylinositol-anchored proteins, aminopeptidase (APN) and alkaline phosphatase (ALP), h...

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

    DEFF Research Database (Denmark)

    Haidl, Felix; Plesner, Torben; Lund, Thomas

    2012-01-01

    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 myelo...

  1. Multiple forms of alkaline phosphatase from Escherichia coli cells with repressed and derepressed biosynthesis of the enzyme.

    OpenAIRE

    Nesmeyanova, M A; Motlokh, O B; Kolot, M N; Kulaev, I. S.

    1981-01-01

    Isolation of multiple forms of alkaline phosphatase from Escherichia coli cells with repressed and derepressed biosynthesis of the enzyme is reported. Three enzyme forms were isolated from cells with derepressed synthesis, and one form was isolated from cells with repressed enzyme synthesis. The multiple enzyme forms did not differ in pH optimum, thermostability, or the degree of inhibition with orthophosphate; however, they did differ in the relative rate of hydrolysis of different substrate...

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

    Science.gov (United States)

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

    2011-08-01

    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

  3. Indirect alkaline phosphatase immunoenzymatic staining for the detection of antibodies to Epstein-Barr virus-induced virus capsid antigens and early antigens.

    OpenAIRE

    Musiani, M.; Zerbini, M.; Gentilomi, G.; La Placa, M.

    1986-01-01

    An indirect alkaline phosphatase immunoenzymatic staining technique was developed for the detection of antibodies against Epstein-Barr virus-induced virus capsid antigens and early antigens in cell smears. The presence of antibodies against Epstein-Barr virus-induced virus capsid antigens and early antigens was revealed by a dark blue staining of cells expressing the antigens. The alkaline phosphatase assay gave a permanent record of the reaction that could be visualized under an ordinary lig...

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

    Scientific Electronic Library Online (English)

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

    2000-08-01

    Full Text Available A constitutive alkaline phosphatase was purified to apparent homogeneity as determined by polyacrylamide gel electrophoresis from mycelia of the wild strain 74A of the mold Neurospora crassa, after growth on acetate and in the presence of saturating amounts of inorganic phosphate (Pi) for 72 h at 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.

  5. Characterization of rat heart alkaline phosphatase isoenzymes and modulation of activity

    Scientific Electronic Library Online (English)

    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

    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.

  6. Recombinant production and characterization of a highly active alkaline phosphatase from marine bacterium Cobetia marina.

    Science.gov (United States)

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

    2015-04-01

    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

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

    Science.gov (United States)

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

    2015-10-20

    The alkaline phosphatase (ALP) biocatalysis followed by the in situ enzymatic generation of a visible light responsive nanozyme is coupled to elucidate a novel amplification strategy by enzymatic cascade reaction for versatile biosensing. The enzymatic hydrolysis of o-phosphonoxyphenol (OPP) to catechol (CA) by ALP is allowed to coordinate on the surface of TiO2 nanoparticles (NPs) due to the specificity and high affinity of enediol ligands to Ti(IV). Upon the stimuli by CA generated from ALP, the inert TiO2 NPs is activated, which demonstrates highly efficient oxidase mimicking activity for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light (? ? 400 nm) irradiation utilizing dissolved oxygen as an electron acceptor. On the basis of the cascade reaction of ALP and the nanozyme of CA coordinated TiO2 (TiO2-CA) NPs, we design exquisitely colorimetric biosensors for probing ALP activity and its inhibitor of 2, 4-dichlorophenoxyacetic acid (2,4-DA). Quantitative probing of ALP activity in a wide linear range from 0.01 to 150 U/L with the detection limit of 0.002 U/L is realized, which endows the methodology with sufficiently high sensitivity for potentially practical applications in real samples of human serum (ALP level of 40-190 U/L for adults). In addition, a novel immunoassay protocol by taking mouse IgG as an example is validated using the ALP/nanozyme cascade amplification reaction as the signal transducer. A low detection limit of 2.0 pg/mL is attained for mouse IgG, which is 4500-fold lower than that of the standard enzyme-linked immuno-sorbent assay (ELISA) kit. Although only mouse IgG is used as a proof-of-concept in our experiment, we believe that this approach is generalizable to be readily extended to other ELISA systems. This methodology opens a new horizon for amplified and versatile biosensing including probing ALP activity and following ALP-based ELISA immunoassays. PMID:26419907

  8. Visualization of alkaline phosphatase-labelled antibodies on immunoblots by means of formazan staining using indoxyl phosphate and thiazolyl blue.

    Science.gov (United States)

    Heegaard, N H

    1990-01-01

    Alkaline phosphatase-conjugated secondary antibodies on blotting membranes were visualized by means of a modified formazan staining method (Hodson & Skillen 1988). It is based on the standard 5-bromo-4-chloro-3-indolyl phosphate/Nitroblue tetrazolium method, but employs different buffer conditions and thiazolyl blue instead of Nitroblue. It gave a sensitivity of 1 ng albumin in a 12 mm2 dot and less background staining on dot blots than the standard method. The new protocol provides an advantage with regard to ease of staining and preparation of reagents while it equals the former method with respect to sensitivity. PMID:1711901

  9. 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)

    Bednyakov Dmitriy Andreevich

    2012-11-01

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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

    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.)

  11. 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

  12. Ultrasensitive detection of amifostine and alkaline phosphatase based on the growth of CdS quantum dots.

    Science.gov (United States)

    Na, Weidan; Liu, Siyu; Liu, Xiaotong; Su, Xingguang

    2015-11-01

    In this study, we reported a simple and sensitive fluorescence nanosensor for rapid detection of amifostine and alkaline phosphatase (ALP). The novel nanosensor was based on the fluorescence "turn on-off" of CdS quantum dots (QDs). Firstly, Cd(2+) cation could react with S(2-) anion to generate fluorescent CdS QDs in the presence of amifostine. The fluorescence (FL) intensity of amifostine-capped CdS QDs (Amifostine-CdS QDs) was increased with the increasing amounts of amifostine, and could be used for amifostine detection. However, amifostine could be converted to 2-(3-aminopropylamino) ethanethiol (WR1065) in the presence of ALP based on the dephosphorylation of ALP. Under the optimum conditions, the affinity of WR1065 to CdS QDs was weaker than that of amifostine. Therefore the new generation of WR1065-CdS QDs would reduce the FL intensity with the increase of ALP concentration, and the fluorescence of CdS QDs was turn off. The metabolic process of amifostine in the presence of alkaline phosphatase could be also studied via the change of FL intensity of CdS QDs. The present method was cost-effective, convenient, and does not require any complicated synthetic procedures. PMID:26452927

  13. Ratio of serum tartrate-inhibitable acid phosphatase to total serum protein in benign prostatic hypertrophy and prostatic carcinoma.

    Science.gov (United States)

    Petrovich, G; Ursich-Jankovich, J; Prijovich, Z

    1992-02-01

    The activity concentration and the specific activity (the ratio of enzyme activity to total serum protein) of the tartrate-inhibitable fraction of acid phosphatase [orthophosphoric monoester phosphohydrolase (acid optimum), EC 3.1.3.2; TIAP] were related to benign prostatic hypertrophy and to prostatic carcinoma. As expected, the TIAP activity concentrations assayed in the sera of patients with benign prostatic hypertrophy were within the range of those assayed in normal human sera. In contrast, the specific activities of TIAP determined in the sera of patients with benign prostatic hypertrophy were significantly higher than those determined in the control group. In the sera of prostatic carcinoma patients, both the TIAP activity concentrations and the TIAP specific activities differed significantly (F = 730) from the normal values. PMID:1371723

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

    International Nuclear Information System (INIS)

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

  15. 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)

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

    2014-09-01

    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

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

    DEFF Research Database (Denmark)

    Haidl, Felix; Plesner, Torben

    2012-01-01

    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.

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

    International Nuclear Information System (INIS)

    Localization of alkaline phosphatase (ALP) and cathepsin D (CAPD) in primary cultures of fetal rat hepatocytes was examined using double immunofluorescent staining in order to investigate the relationship between lysosome movement and the fate of ALP during cell restoration after microtubule disruption by colchicine. At 3 hr and 24 hr after colchicine treatment, numerous coarse dots containing ALP were observed throughout the cytoplasm, and some of these showed colocalization with CAPD. At 48 hr and 72 hr after colchicine treatment, although most of the dots containing ALP in the cytoplasm disappeared, dots containing CAPD remained. The present results suggest that the denatured ALP proteins remaining in the cytoplasm of hepatocytes during cell restoration after colchicine treatment are digested by lysosomes

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

    Science.gov (United States)

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

    2015-05-01

    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

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

    Science.gov (United States)

    Liu, Jia-Ming; Huang, Xiao-Mei; Liu, Zhen-Bo; Lin, Shao-Qin; Li, Fei-Ming; Gao, Fei; Li, Zhi-Ming; Zeng, Li-Qing; Li, Lian-Ying; Ouyang, Ying

    2009-08-26

    A new phosphorescent labelling reagent consisting of fullerol, fluorescein isothiocyanate and N,N-dimethylaniline (F-ol-(FITC)(n)-DMA) was developed. The mode of action is based on the reactivity of the active -OH group in F-ol with the -COOH group of FITC to form an F-ol-(FITC)(n)-DMA complex containing several FITC molecules. F-ol-(FITC)(n)-DMA increased the number of luminescent molecules in the biological target of WGA-AP-WGA-F-ol-(FITC)(n)-DMA (WGA and AP are wheat germ agglutinin and alkaline phosphatase, respectively) which improved the sensitivity using solid substrate room temperature phosphorimetry (SSRTP) detection. The proposed method provided high sensitivity and strong specificity for WGA-AP. The limit of detection (LD) was 0.15 ag AP spot(-1) for F-ol and 0.097 ag AP spot(-1) for FITC in F-ol-(FITC)(n)-DMA, which was lower than the method using single luminescent molecules of F-ol-DMA and FITC-DMA to label WGA (0.20 ag AP spot(-1) for F-ol-DMA and 0.22 ag AP spot(-1) for FITC-DMA). Results for the determination of AP in human serum were in good agreement with those obtained by enzyme-linked immunosorbent assay. The mechanism of F-ol-(FITC)(n)-DMA labelling of WGA was discussed. PMID:19646588

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

    International Nuclear Information System (INIS)

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

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

    Science.gov (United States)

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

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

    OpenAIRE

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

    2010-01-01

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

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

    International Nuclear Information System (INIS)

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

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

    Scientific Electronic Library Online (English)

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

    2012-03-01

    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+.

  5. 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

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

    Directory of Open Access Journals (Sweden)

    Grozdea Jean J

    2002-01-01

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

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

    International Nuclear Information System (INIS)

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

  8. 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

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

    Directory of Open Access Journals (Sweden)

    Bertha B Socarrás Ferrer

    2006-04-01

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

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

    Scientific Electronic Library Online (English)

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

    2006-04-01

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

  11. Alkaline phosphatase activity at the southwest coast of India: A comparison of locations differently affected by upwelling

    Science.gov (United States)

    Mamatha, S. S.; Malik, Ashish; Varik, Sandesh; Parvathi, V.; Jineesh, V. K.; Gauns, Mangesh U.; LokaBharathi, P. A.

    2015-01-01

    The realization of the potential importance of phosphorus (P) as a limiting nutrient in marine ecosystem is increasing globally. Hence, the contribution of biotic variables in mobilizing this nutrient would be relevant especially in productive coastal waters. As alkaline phosphatase activity (APA) indicates the status of P for primary production in aquatic environments, we asked the following question: is the level of APA indicative of P sufficiency or deficiency in coastal waters, especially, where upwelling is a regular phenomenon? Therefore, we have examined the total APA, chlorophyll a along with phosphatase producing bacteria (PPB) and related environmental parameters from nearshore to offshore in coastal waters off Trivandrum and Kochi regions differently affected by upwelling during the onset of monsoon. Off Trivandrum, APA in the offshore waters of 5-m layer at 2.23 ?M P h- 1 was > 4 times higher than nearshore. Thus, low APA could be indicative of P sufficiency in coastal waters and higher activity suggestive of deficiency in offshore waters off Trivandrum. In contrast, there was less difference in APA between near and offshore surface waters off Kochi. Our results show that the regions differently affected by upwelling respond differently according to ambient P concentration, distance from shore or depth of water. These observations could apparently be applicable to other coastal systems as well, where gradients in upwelling and phosphate runoff have been noticed. Further studies on other transects would throw more light on the extent and direction of the relationship between APA and ambient P concentration. Such studies would help in understanding the level of control of this nutrient on the productivity of coastal waters.

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

    Directory of Open Access Journals (Sweden)

    Henk-Jan Prins

    2014-03-01

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

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

    Science.gov (United States)

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

    2003-01-01

    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.

  14. 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 may arise from complementarity of the guanidinum group to the geometry of the trigonal bipyramidal transition state

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

    Directory of Open Access Journals (Sweden)

    Almohamad Khaled M.

    2014-08-01

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

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

    Science.gov (United States)

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

    2010-05-01

    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

  17. Change in Localization of Alkaline Phosphatase and Mannosidase II by Colchicine Treatment of Primary Cultures of Fetal Rat Hepatocytes

    International Nuclear Information System (INIS)

    We examined the changes in localization of alkaline phosphatase (ALP) and mannosidase II (man II), a Golgi marker, after colchicine treatment of primary cultures of fetal rat hepatocytes, using double immunofluorescence staining and confocal laser microscopy. In hepatocytes cultured in basal medium, ALP was localized in the perinuclear cytoplasm, and man II was observed in the Golgi region of the cytoplasm. When hepatocytes were cultured in dexamethasone-supplemented medium, ALP was also localized in the plasma membrane surrounding the bile canaliculus-like structure that was formed between adjacent cells. In hepatocytes cultured in the same medium containing colchicine, the structure of microtubules in the cytoplasm was lost, man II exhibited granular distribution scattering throughout the cytoplasm, and ALP was localized in coarse granular sites of the cytoplasm. However, ALP was not colocalized at the same sites as man II. The present study indicated that colchicine inhibits the dexamethasone-promoted translocation of ALP to the plasma membrane surrounding the bile canaliculus-like structure in primary cultures of fetal rat hepatocytes by disassembling microtubules and discomposing the Golgi complex

  18. Effects of Colchicine on Localization of Alkaline Phosphatase in McA-RH 7777 Rat Hepatoma Cells

    International Nuclear Information System (INIS)

    We investigated the changes caused by microtubule disruption in cell contact-induced translocation of alkaline phosphatase (ALP) from the Golgi area to the plasma membrane in McA-RH 7777 cells. When the cells were treated with colchicine, the tubular structure of microtubules in the cytoplasm was lost. Colchicine treatment also resulted in the appearance of numerous dots containing mannosidase II (man II) throughout the cytoplasm. Moreover, ALP was distributed in small dots throughout the cytoplasm, as well as in all regions of the plasma membrane, although it was most concentrated at sites of intercellular contact. On the other hand, when the cells were incubated in basal medium after colchicine treatment, large spots containing ALP reappeared in the perinuclear cytoplasm more quickly than the accumulation of small dots containing man II. These findings suggest that colchicine causes disassembly of the Golgi complex into fragments, which scatter throughout the cytoplasm, but that it does not interfere with translocation of ALP to the plasma membrane. Furthermore, cytoplasmic ALP may be localized at sites other than the Golgi complex

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

    Energy Technology Data Exchange (ETDEWEB)

    Liu Jiaming, E-mail: zzsyliujiaming@163.com [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000 (China); Huang Xiaomei [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000 (China); Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000 (China); Liu Zhenbo [Third Hospital of Xiamen, Xiamen 361100 (China); Lin Shaoqin [Department of Biochemistry, Fujian Education College, Fuzhou 350001 (China); Li Feiming; Gao Fei [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000 (China); Li Zhiming [Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000 (China); Zeng Liqing; Li Lianying; Ouyang Ying [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000 (China)

    2009-08-26

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

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    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...

  1. Synthesis of 2'(3')-O-DL-alanyl hexainosinic acid using T4 RNA ligase: suppression of the enzymic reverse transfer reaction by alkaline phosphatase.

    Science.gov (United States)

    Profy, A T; Lo, K M; Usher, D A

    1983-01-01

    2'(3')-O-DL-Alanyl (Ip)5I was synthesized by a new method. An alanine ortho ester of inosine 5'-phosphate was added to (Ip)4I using the ATP-independent reaction of T4 RNA ligase, and the product was converted smoothly to the desired ester. The enzymic reverse transfer reaction was conveniently suppressed by the dephosphorylation of the adenosine 5'-phosphate coproduct, catalyzed in situ by alkaline phosphatase. PMID:6857753

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

    OpenAIRE

    G.E. Eriyamremu; M.A. Adaikpoh; F.O. Obi

    2006-01-01

    Cell membrane composition and fluidity are altered in diseases and previous reports suggest that membrane lipid is altered in heavy metal toxicity. This study was carried out to assess the effect of cadmium alone and its combination with different doses (75, 150 and 750 mg) of ?-tocophenylacetate on the phospholipid profile and alkaline phosphatase activity in the kidney and liver of rats. The results obtained show that in these tissues, cadmium significantly (p<0.05) increased the levels ...

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

    International Nuclear Information System (INIS)

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

  4. 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)

    OpenAIRE

    Dobado-Berrios, P.M.; Ferrer, Miguel, n. 1526

    1997-01-01

    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 ...

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

    OpenAIRE

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

    2011-01-01

    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 ...

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

    Directory of Open Access Journals (Sweden)

    Zoraida Pons Pinillos

    2005-08-01

    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.

  7. Amplified thrombin aptasensor based on alkaline phosphatase and hemin/G-quadruplex-catalyzed oxidation of 1-naphthol.

    Science.gov (United States)

    Yang, Zhe-Han; Zhuo, Ying; Yuan, Ruo; Chai, Ya-Qin

    2015-05-20

    An alkaline phosphatase (ALP)-based biosensor can in situ generate an electroactive product by enzymatic hydrolysis of inactive substrates. To obtain a higher signal-to-background ratio, a chemical redox cycling signal-amplified strategy based on the addition of a strong reducing agent has often be applied in the construction of ALP-based biosensors. However, the strong reducing agent not only affects the activity of ALP but also readily reacts with dissolved oxygen, leading to inaccurate results. In this work, a new signal-amplified strategy for a thrombin (TB) aptasensor based on the catalytic oxidation of ALP-generated products, 1-naphthol (NP), using hemin/G-quadruplex DNAzymes was reported. We implemented gold-nanoparticle-decorated zinc oxide nanoflowers (Au-ZnO) as the matrix for immobilizing ALP and TB aptamer (TBA) and then labeled it with hemin to form hemin/G-quadruplex/ALP/Au-ZnO bioconjugates (TBA II bioconjugates). Through a "sandwich" reaction, TBA II bioconjugates were captured on the electrode surface. The amplified signal was carried out in two steps: (i) an ALP-catalyzed inactive substrate, 1-naphthyl phosphate (NPP), in situ produces NP on the surface of the electrode; (ii) on the one hand, NP as a new reactant could be directly electrooxidized and generated an electrochemical signal, but, on the other hand, NP could be oxidized by hemin/G-quadruplex in the presence of H2O2, resulting in amplification of the electrochemical signal. The proposed TB aptasensor achieved a linear range of 1 pM to 30 nM with a detection limit of 0.37 pM (defined as S/N = 3). PMID:25907268

  8. Pueraria mirifica extract and puerarin enhance proliferation and expression of alkaline phosphatase and type I collagen in primary baboon osteoblasts.

    Science.gov (United States)

    Tiyasatkulkovit, Wacharaporn; Malaivijitnond, Suchinda; Charoenphandhu, Narattaphol; Havill, Lorena M; Ford, Allen L; VandeBerg, John L

    2014-10-15

    Phytoestrogen-rich Pueraria mirifica (PM) tuberous extract is a promising candidate for the development of anti-osteoporosis drugs for postmenopausal women, but its action has never been validated in humans or in non-human primates, which are more closely related to humans than rodents. In vitro study of non-human primate osteoblasts is thus fundamental to prepare for in vivo studies of phytoestrogen effects on primate bone. This study aimed to establish a culture system of baboon primary osteoblasts and to investigate the effects of PM extract and its phytoestrogens on these cells. Primary osteoblasts from adult baboon fibulae exhibited osteoblast characteristics in regard to proliferation, differentiation, mineralization, and estrogen receptor expression. They responded to 17?-estradiol by increased proliferation rate and mRNA levels of alkaline phosphatase (ALP), type I collagen, and osteocalcin. After being exposed for 48 h to 100 ?g/ml PM extract, 1000 nM genistein, or 1000 nM puerarin, primary baboon osteoblasts markedly increased the rate of proliferation and mRNA levels of ALP and type I collagen without changes in Runx2, osterix, or osteocalcin expression. PM extract, genistein, and puerarin also decreased the RANKL/OPG ratio, suggesting that they could decrease osteoclast-mediated bone resorption. However, neither PM extract nor its phytoestrogens altered calcium deposition in osteoblast culture. In conclusion, we have established baboon primary osteoblast culture, which is a new tool for bone research and drug discovery. Furthermore, the present results provide substantial support for the potential of PM extract and its phytoestrogens to be developed as therapeutic agents against bone fragility. PMID:25442257

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

    Scientific Electronic Library Online (English)

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

    2008-01-01

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

  10. Alkaline phosphatase fusions to the respiratory syncytial virus F protein as an approach to analyze its membrane topology.

    Science.gov (United States)

    Martin-Gallardo, A; Deich, R A; Fien, K A; Metcalf, B J; Anilionis, A; Paradiso, P R

    1989-11-01

    Manoil and Beckwith (1985) have constructed a transposon, TnphoA, that permits the generation of hybrid proteins composed of alkaline phosphatase (AP) lacking its signal peptide fused to amino-terminal sequences of other proteins. This transposon has been used to localize export signals and analyze membrane topology of bacterial proteins. We have applied this approach to the membrane fusion protein (F) of respiratory syncytial virus (RSV). The transposon TnphoA and a plasmid directing bacterial expression of the F gene were used to construct F-AP hybrids. These hybrids yielded AP activity, indicating the presence of viral sequences that promoted protein transport through the cytoplasmic membrane. Sequence analysis showed that TnphoA was inserted at four different positions within the F1 subunit. Deletion of the hydrophobic F1 amino-terminus (fusion-related domain) resulted in AP transport to the periplasm, suggesting that the hydrophobic amino-terminus of the F2 subunit is sufficient to promote protein export. Some hybrids were apparently cleaved at or near the F2/F1 junction. The periplasmic localization of an uncleaved hybrid strongly suggested that the fusion-related domain of the F protein, when in the uncleaved F0 precursor, can be moved across the bacterial cytoplasmic membrane. Although these results apply to the recombinant F protein, they agree with the presumed signal sequence and membrane topology of the native F glycoprotein. Thus, this method may be useful in determining membrane topology and in localizing important domains of viral proteins. PMID:2558867

  11. A Rapid Lateral Flow Immunoassay for the Detection of Tyrosine Phosphatase-Like Protein IA-2 Autoantibodies in Human Serum

    OpenAIRE

    Kikkas, Ingrid; Mallone, Roberto; Larger, Etienne; Volland, Hervé; Morel, Nathalie

    2014-01-01

    Type 1 diabetes (T1D) results from the destruction of pancreatic insulin-producing beta cells and is strongly associated with the presence of islet autoantibodies. Autoantibodies to tyrosine phosphatase-like protein IA-2 (IA-2As) are considered to be highly predictive markers of T1D. We developed a novel lateral flow immunoassay (LFIA) based on a bridging format for the rapid detection of IA-2As in human serum samples. In this assay, one site of the IA-2As is bound to HA-tagged-IA-2, which is...

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

    Directory of Open Access Journals (Sweden)

    Peifang Wang

    2014-08-01

    Full Text Available Phosphorus (P is an important limiting nutrient in aquatic ecosystems and knowledge of P cycling is fundamental for reducing harmful algae blooms and other negative effects in water. Despite their importance, the characteristics of P cycling under changing nutrient conditions in shallow lakes were poorly investigated. In this study, in situ incubation experiments were conducted in a natural riparian zone in the main diversion channel used for water transfer into Lake Taihu (Wangyu River. Variations in microbial biomass, dissolved P fractions (organic and inorganic, and alkaline phosphatase activity (bulk APA and specific APA were determined after incubation with and without the addition of P and nitrogen (N (4 total water treatments: +P, +N, +NP, and control. Experiments were conducted during two seasons (late spring and early fall to account for natural differences in nutrient levels that may occur in situ. Our results demonstrated that low levels of DRP may not necessarily indicate P limitation. Phytoplankton exhibited “serial N limitation with P stress” in May, such that chlorophyll a (Chl a increased significantly with N addition, while the limiting nutrient shifted to P in October and phytoplankton biomass increased with P addition. Phytoplankton contributed greatly to APA production and was significantly influenced by P bioavailability, yet high levels of bulk APA were also not necessarily indicative of P limitation. In contrast to phytoplankton, bacteria were less P stressed. As a consequence of enhanced utilization of dissolved reactive P (DRP and dissolved organic P (DOP, +N treatment elevated APA significantly. By contrast, APA could be repressed to low values and phytoplankton converted a large portion of DRP to DOP with P addition. But this was not consistent with bacteria APA (bact-APA in the absence or presence of abundant phytoplankton biomass. The correlation between bulk APA and DRP was good at separate sites and discrepant for the whole data set. Regulation of APA was demonstrated by an inverse hyperbolic relationship between bulk APA, specific APA, and DRP, with a transition from high to low activity occurring between 20 and 50 ?g L-1. This study provides a better understanding of how APA and P cycling change with nutrient perturbations in Lake Taihu system. The obtained results can help understanding the process of P cycling in water and providing a reference for nutrient control in the water transfer project.

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

    DEFF Research Database (Denmark)

    Ornstrup, Marie Juul; HarslØf, Torben

    2014-01-01

    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.

  14. Subcellular localisation and properties of histone phosphate phosphatase in human polymorphonuclear leukocytes: alterations in pregnancy and chronic granulocytic leukaemia and relationship to alkaline phosphatase

    International Nuclear Information System (INIS)

    Using [32P]histone as substrate, an assay for histone phosphate phosphatase was optimised for human polymorphonuclear leukocytes. Kinetic studies showed that the activity was optimal at pH 6.8, was stimulated by Mn2+ and Co2+, and inhibited by sodium sulphite and zinc chloride. The apparent Ksub(m) of the enzyme for histone phosphate was 0.89 ?mol/l. (Auth.)

  15. Changes in endonuclease activity with acid and alkaline pH in rat blood serum after whole-body ?-irradiation

    International Nuclear Information System (INIS)

    Using the method of electrophoresis in agar gel of superhelical plasmid DNA, which served as a substrate for determining endonuclease activity, the authors showed that the same radiation dose influenced in a different way the activity of acid and alkaline DNAases in rat blood serum. At early times (3-6 h) following irradiation, the activity of acid nucleases increaed whereas that of alkaline DNAases decreased

  16. Escherichia coli alkaline phosphatase: X-ray structural studies of a mutant enzyme (His-412-->Asn) at one of the catalytically important zinc binding sites.

    OpenAIRE

    Ma, L.; Tibbitts, T. T.; Kantrowitz, E R

    1995-01-01

    The X-ray structure of a mutant version of Escherichia coli alkaline phosphatase (H412N) in which His-412 was replaced by Asn has been determined at both low (-Zn) and high (+Zn) concentrations of zinc. In the wild-type structure, His-412 is a direct ligand to one of the two catalytically critical zinc atoms (Zn1) in the active site. Characterization of the H412N enzyme in solution revealed that the mutant enzyme required high concentrations of zinc for maximal activity and for high substrate...

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

    OpenAIRE

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

    2012-01-01

    Bacillus thuringiensis subs. israelensis produces at least three Cry toxins (Cry4Aa, Cry4Ba, and Cry11Aa) that are active against Aedes aegypti larvae. Previous work characterized a GPI-anchored alkaline phosphatase (ALP1) as a Cry11Aa binding molecule from the gut of A. aegypti larvae. We show here that Cry4Ba binds ALP1, and that the binding and toxicity of Cry4Ba mutants located in loop 2 of domain II is correlated. Also, we analyzed the contribution of ALP1 towards the toxicity of Cry4Ba ...

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

    Directory of Open Access Journals (Sweden)

    Keyhani doost Z

    2011-01-01

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

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

    OpenAIRE

    Liu Hsin-Yi; Lee Su-Hui; Yu Jyh-Cherng; Yu Cheng-Ping; Ku Chih-Hung; Janckila Anthony J; Wu Yi-Ying; Yam Lung T; Chao Tsu-Yi

    2010-01-01

    Abstract Background Serum tartrate-resistant acid phosphatase 5b (TRACP 5b) activity is a marker of osteoclast number and is elevated in breast cancer (BC) patients with extensive bone metastasis, which might in turn reflect the tumour burden. We tested the hypothesis that baseline serum TRACP 5b activity and its interval change are potential prognostic markers of survival in BC patients with bone metastasis. Methods We analyzed the data from previous prospective studies. A total of 100 patie...

  20. 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)

  1. 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)

    Bajin-Kati? Katica

    2006-01-01

    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.

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

    International Nuclear Information System (INIS)

    Graphical abstract: A phage-displayed chicken scFv antibody, FvSG7, binds on the surface antigen of conidiospores and the mycelia of F. verticillioides. Its fusion with alkaline phosphatase (AP) through a 218 linker displayed a 4-fold higher affinity compared with the parent scFv antibody and efficiently detected toxigenic Fusarium pathogens in cereal grains. Highlights: ? Generation of a highly reactive scFv antibody against F. verticillioides. ? Localization of the antibody binding to the surface target of F. verticillioides. ? Expression of the antibody–alkaline phosphatase (AP) fusion linked by a 218 linker. ? The antibody–AP fusion has a higher affinity than the parental antibody. ? The antibody–AP fusion detects toxigenic Fusarium pathogens in cereal grains. -- Abstract: Fusarium and its poisonous mycotoxins are distributed worldwide and are of particular interest in agriculture and food safety. A simple analytical method to detect pathogens is essential for forecasting diseases and controlling mycotoxins. This article describes a proposed method for convenient and sensitive detection of Fusarium pathogens that uses the fusion of single-chain variable fragment (scFv) and alkaline phosphatase (AP). A highly reactive scFv antibody specific to soluble cell wall-bound proteins (SCWPs) of F. verticillioides was selected from an immunized chicken phagemid library by phage display. The antibody was verified to bind on the surface of ungerminated conidiospores and mycelia of F. verticillioides. The scFv–AP fusion was constructed, and soluble expression in bacteria was confirmed. Both the antibody properties and enzymatic activity were retained, and the antigen-binding capacity of the fusion was enhanced by the addition of a linker. Surface plasmon resonance measurements confirmed that the fusion displayed 4-fold higher affinity compared with the fusion's parental scFv antibody. Immunoblot analyses showed that the fusion had good binding capacity to the components from SCWPs of F. verticillioides, and enzyme-linked immunosorbent assays revealed that the detection limit of the fungus was below 10?2 ?g mL?1, superior to the scFv antibody. The fusion protein was able to detect fungal concentrations as low as 10?3 mg g?1 of maize grains in both naturally and artificially contaminated samples. Thus, the fusion can be applied in rapid and simple diagnosis of Fusarium contamination in field and stored grain or in food

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

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Zu-Quan [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Li, He-Ping [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Zhang, Jing-Bo [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Huang, Tao [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); Liu, Jin-Long; Xue, Sheng [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); Wu, Ai-Bo [Institute for Agri-food Standards and Testing Technology, Laboratory of Quality and Safety Risk Assessment for Agro-products, Ministry of Agriculture, Shanghai Academy of Agricultural Sciences, 1000 Jinqi Road, Shanghai 201403 (China); Liao, Yu-Cai, E-mail: ycliao06@yahoo.com.cn [Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan 430070 (China); College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070 (China); National Center of Plant Gene Research, Wuhan 430070 (China)

    2013-02-18

    Graphical abstract: A phage-displayed chicken scFv antibody, FvSG7, binds on the surface antigen of conidiospores and the mycelia of F. verticillioides. Its fusion with alkaline phosphatase (AP) through a 218 linker displayed a 4-fold higher affinity compared with the parent scFv antibody and efficiently detected toxigenic Fusarium pathogens in cereal grains. Highlights: ? Generation of a highly reactive scFv antibody against F. verticillioides. ? Localization of the antibody binding to the surface target of F. verticillioides. ? Expression of the antibody–alkaline phosphatase (AP) fusion linked by a 218 linker. ? The antibody–AP fusion has a higher affinity than the parental antibody. ? The antibody–AP fusion detects toxigenic Fusarium pathogens in cereal grains. -- Abstract: Fusarium and its poisonous mycotoxins are distributed worldwide and are of particular interest in agriculture and food safety. A simple analytical method to detect pathogens is essential for forecasting diseases and controlling mycotoxins. This article describes a proposed method for convenient and sensitive detection of Fusarium pathogens that uses the fusion of single-chain variable fragment (scFv) and alkaline phosphatase (AP). A highly reactive scFv antibody specific to soluble cell wall-bound proteins (SCWPs) of F. verticillioides was selected from an immunized chicken phagemid library by phage display. The antibody was verified to bind on the surface of ungerminated conidiospores and mycelia of F. verticillioides. The scFv–AP fusion was constructed, and soluble expression in bacteria was confirmed. Both the antibody properties and enzymatic activity were retained, and the antigen-binding capacity of the fusion was enhanced by the addition of a linker. Surface plasmon resonance measurements confirmed that the fusion displayed 4-fold higher affinity compared with the fusion's parental scFv antibody. Immunoblot analyses showed that the fusion had good binding capacity to the components from SCWPs of F. verticillioides, and enzyme-linked immunosorbent assays revealed that the detection limit of the fungus was below 10{sup ?2} ?g mL{sup ?1}, superior to the scFv antibody. The fusion protein was able to detect fungal concentrations as low as 10{sup ?3} mg g{sup ?1} of maize grains in both naturally and artificially contaminated samples. Thus, the fusion can be applied in rapid and simple diagnosis of Fusarium contamination in field and stored grain or in food.

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

    DEFF Research Database (Denmark)

    Lisse, I M; Whittle, H

    1990-01-01

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

  5. Chitosan nanofiber scaffold improves bone healing via stimulating trabecular bone production due to upregulation of the Runx2/osteocalcin/alkaline phosphatase signaling pathway

    Science.gov (United States)

    Ho, Ming-Hua; Yao, Chih-Jung; Liao, Mei-Hsiu; Lin, Pei-I; Liu, Shing-Hwa; Chen, Ruei-Ming

    2015-01-01

    Osteoblasts play critical roles in bone formation. Our previous study showed that chitosan nanofibers can stimulate osteoblast proliferation and maturation. This translational study used an animal model of bone defects to evaluate the effects of chitosan nanofiber scaffolds on bone healing and the possible mechanisms. In this study, we produced uniform chitosan nanofibers with fiber diameters of approximately 200 nm. A bone defect was surgically created in the proximal femurs of male C57LB/6 mice, and then the left femur was implanted with chitosan nanofiber scaffolds for 21 days and compared with the right femur, which served as a control. Histological analyses revealed that implantation of chitosan nanofiber scaffolds did not lead to hepatotoxicity or nephrotoxicity. Instead, imaging analyses by X-ray transmission and microcomputed tomography showed that implantation of chitosan nanofiber scaffolds improved bone healing compared with the control group. In parallel, microcomputed tomography and bone histomorphometric assays further demonstrated augmentation of the production of new trabecular bone in the chitosan nanofiber-treated group. Furthermore, implantation of chitosan nanofiber scaffolds led to a significant increase in the trabecular bone thickness but a reduction in the trabecular parameter factor. As to the mechanisms, analysis by confocal microscopy showed that implantation of chitosan nanofiber scaffolds increased levels of Runt-related transcription factor 2 (Runx2), a key transcription factor that regulates osteogenesis, in the bone defect sites. Successively, amounts of alkaline phosphatase and osteocalcin, two typical biomarkers that can simulate bone maturation, were augmented following implantation of chitosan nanofiber scaffolds. Taken together, this translational study showed a beneficial effect of chitosan nanofiber scaffolds on bone healing through stimulating trabecular bone production due to upregulation of Runx2-mediated alkaline phosphatase and osteocalcin gene expressions. Our results suggest the potential of chitosan nanofiber scaffolds for therapy of bone diseases, including bone defects and bone fractures. PMID:26451104

  6. 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)

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

    2011-12-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Alvarez M

    2008-04-01

    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.

  8. Ecto-alkaline phosphatase activity identified at physiological pH range on intact P19 and HL-60 cells is induced by retinoic acid.

    Science.gov (United States)

    Scheibe, R J; Kuehl, H; Krautwald, S; Meissner, J D; Mueller, W H

    2000-01-01

    The activity of membrane-bound alkaline phosphatase (ALP) expressed on the external surface of cultured murine P19 teratocarcinoma and human HL-60 myeloblastic leukemia cells was studied at physiological pH using p-nitrophenylphosphate (pNPP) as substrate. The rate of substrate hydrolysis catalyzed by intact viable cells remained constant for eight successive incubations of 30 min and was optimal at micromolar substrate concentrations over the pH range 7.4-8.5. The value of apparent K(m) for pNPP in P19 and HL-60 cells was 120 microM. Hydrolytic activity of the ecto-enzyme at physiological pH decreased by the addition of levamisole, a specific and noncompetitive inhibitor of ALP (K(i) P19 = 57 microM; K(i) HL-60 = 50 microM). Inhibition of hydrolysis was reversed by removal of levamisole within 30 min. Retinoic acid (RA), which promotes the differentiation of P19 and HL-60 cells, induced levamisole-sensitive ecto-phosphohydrolase activity at pH 7.4. After its autophosphorylation by ecto-kinase activity, a 98-kDa membrane protein in P19 cells was found to be sensitive to ecto-ALP, and protein dephosphorylation increased after incubation of cells with RA for 24 h and 48 h. Orthovanadate, an inhibitor of all phosphatase activities, blocked the levamisole-sensitive dephosphorylation of the membrane phosphoproteins, while (R)-(-)-epinephrine reversed the effect by complexation of the inhibitor. The results demonstrate that the levamisole-sensitive phosphohydrolase activity on the cell surface is consistent with ecto-ALP activity degrading both physiological concentrations of exogenously added substrate and endogenous surface phosphoproteins under physiological pH conditions. The dephosphorylating properties of ecto-ALP are induced by RA, suggesting a specific function in differentiating P19 teratocarcinoma and HL-60 myeloblastic leukemia cells. PMID:10649440

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

    Directory of Open Access Journals (Sweden)

    Kala Mrinalini

    2005-12-01

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1990-04-01

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

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

    Science.gov (United States)

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

    2015-07-01

    Monitoring the effects of no-tillage (NT) in comparison with conventional tillage (CT) on soil microbes could improve our understanding of soil biochemical processes and thus help us to develop sound management strategies. The objective of this study was to compare the species composition and ecological function of soil arbuscular mycorrhizal (AM) fungi during the growth and rotation of crops under NT and CT. From late June 2009 to early June 2010, 32 topsoil (0-15 cm) samples from four individual plots per treatment (CT and NT) were collected at both the jointing and maturation stages of maize (Zea mays L.) and wheat (Triticum aestivum L.) from a long-term experimental field that was established in an Aquic Inceptisol in North China in June 2006. The AM fungal spores were isolated and identified and then used to calculate species diversity indices, including the Shannon- Wiener index (H'), Evenness (E), and Simpson's index (D). The root mycorrhizal colonization and soil alkaline phosphatase activity were also determined. A total of 34 species of AM fungi within nine genera were recorded. Compared with NT, CT negatively affected the soil AM fungal community at the maize sowing stage, leading to decreases in the average diversity indices (from 2.12, 0.79, and 0.82 to 1.79, 0.72, and 0.74 for H', E, and D, respectively), root mycorrhizal colonization (from 28% to 20%), soil alkaline phosphatase activity (from 0.24 to 0.19 mg/g/24 h) and available phosphorus concentration (from 17.4 to 10.5 mg/kg) at the maize jointing stage. However, reductions in diversity indices of H', E, and D were restored to 2.20, 0.81, and 0.84, respectively, at the maize maturation stage. CT should affect the community again at the wheat sowing stage; however, a similar restoration in the species diversity of AM fungi was completed before the wheat jointing stage, and the highest Jaccard index (0.800) for similarity in the species composition of soil AM fungi between CT and NT was recorded at the wheat maturation stage. Our results also demonstrated that NT resulted in the positive protection of the community structure of AM fungi and played an important role in maintaining their functionality especially for maize seedlings. PMID:26115994

  12. Chitosan nanofiber scaffold improves bone healing via stimulating trabecular bone production due to upregulation of the Runx2/osteocalcin/alkaline phosphatase signaling pathway

    Directory of Open Access Journals (Sweden)

    Ho MH

    2015-09-01

    Full Text Available Ming-Hua Ho,1,2 Chih-Jung Yao,3 Mei-Hsiu Liao,4 Pei-I Lin,4 Shing-Hwa Liu,5 Ruei-Ming Chen2,4,6 1Department of Chemical Engineering, National Taiwan University of Science and Technology, 2Cell Physiology and Molecular Image Research Center, Taipei Medical University-Wan Fang Hospital, 3Department of Internal Medicine, School of Medicine, 4Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, 5Institute of Toxicology, College of Medicine, National Taiwan University, 6Anesthetics and Toxicology Research Center, Taipei Medical University Hospital, Taipei, Taiwan Abstract: Osteoblasts play critical roles in bone formation. Our previous study showed that chitosan nanofibers can stimulate osteoblast proliferation and maturation. This translational study used an animal model of bone defects to evaluate the effects of chitosan nanofiber scaffolds on bone healing and the possible mechanisms. In this study, we produced uniform chitosan nanofibers with fiber diameters of approximately 200 nm. A bone defect was surgically created in the proximal femurs of male C57LB/6 mice, and then the left femur was implanted with chitosan nanofiber scaffolds for 21 days and compared with the right femur, which served as a control. Histological analyses revealed that implantation of chitosan nanofiber scaffolds did not lead to hepatotoxicity or nephrotoxicity. Instead, imaging analyses by X-ray transmission and microcomputed tomography showed that implantation of chitosan nanofiber scaffolds improved bone healing compared with the control group. In parallel, microcomputed tomography and bone histomorphometric assays further demonstrated augmentation of the production of new trabecular bone in the chitosan nanofiber-treated group. Furthermore, implantation of chitosan nanofiber scaffolds led to a significant increase in the trabecular bone thickness but a reduction in the trabecular parameter factor. As to the mechanisms, analysis by confocal microscopy showed that implantation of chitosan nanofiber scaffolds increased levels of Runt-related transcription factor 2 (Runx2, a key transcription factor that regulates osteogenesis, in the bone defect sites. Successively, amounts of alkaline phosphatase and osteocalcin, two typical biomarkers that can simulate bone maturation, were augmented following implantation of chitosan nanofiber scaffolds. Taken together, this translational study showed a beneficial effect of chitosan nanofiber scaffolds on bone healing through stimulating trabecular bone production due to upregulation of Runx2-mediated alkaline phosphatase and osteocalcin gene expressions. Our results suggest the potential of chitosan nanofiber scaffolds for therapy of bone diseases, including bone defects and bone fractures. Keywords: chitosan nanofibers, bone healing, micro-computed tomography, bone histomorphometry, Runx2/OCN/ALP

  13. 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)

    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

    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

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

    Directory of Open Access Journals (Sweden)

    Benjamin Nnamdi Okolonkwo

    2013-06-01

    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.

  15. 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)

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

    2011-03-01

    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

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

    DEFF Research Database (Denmark)

    Rangnoi, Kuntalee; Jaruseranee, Nanthnit

    2011-01-01

    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.

  17. 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)

    Yang YU

    2011-02-01

    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.

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

    Science.gov (United States)

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

    2015-08-01

    Lack of enteral nutrition (EN) during parenteral nutrition (PN) leads to higher incidence of infection because of gut barrier dysfunction. However, the effects of partial EN on intestina linnate immunity, intestinal alkaline phosphatase (IAP) and microbiota remain unclear. The mice were randomized into six groups to receive either standard chow or isocaloric and isonitrogenous nutritional support with variable partial EN to PN ratios. Five days later, the mice were sacrificed and tissue samples were collected. Bacterial translocation, the levels of lysozyme, mucin 2 (MUC2), and IAP were analyzed. The composition of intestinal microbiota was analyzed by 16S rRNA pyrosequencing. Compared with chow, total parenteral nutrition (TPN) resulted in a dysfunctional mucosal barrier, as evidenced by increased bacterial translocation (p TPN group (p < 0.001). These changes were accompanied by maintained barrier function in bacterial culture (p < 0.05). Supplementation of PN with 20% EN preserves gut barrier function, by way of maintaining innate immunity, IAP and intestinal microbiota. PMID:26247961

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

    Directory of Open Access Journals (Sweden)

    Xiao Wan

    2015-08-01

    Full Text Available Lack of enteral nutrition (EN during parenteral nutrition (PN leads to higher incidence of infection because of gut barrier dysfunction. However, the effects of partial EN on intestina linnate immunity, intestinal alkaline phosphatase (IAP and microbiota remain unclear. The mice were randomized into six groups to receive either standard chow or isocaloric and isonitrogenous nutritional support with variable partial EN to PN ratios. Five days later, the mice were sacrificed and tissue samples were collected. Bacterial translocation, the levels of lysozyme, mucin 2 (MUC2, and IAP were analyzed. The composition of intestinal microbiota was analyzed by 16S rRNA pyrosequencing. Compared with chow, total parenteral nutrition (TPN resulted in a dysfunctional mucosal barrier, as evidenced by increased bacterial translocation (p < 0.05, loss of lysozyme, MUC2, and IAP, and changes in the gut microbiota (p < 0.001. Administration of 20% EN supplemented with PN significantly increased the concentrations of lysozyme, MUC2, IAP, and the mRNA levels of lysozyme and MUC2 (p < 0.001. The percentages of Bacteroidetes and Tenericutes were significantly lower in the 20% EN group than in the TPN group (p < 0.001. These changes were accompanied by maintained barrier function in bacterial culture (p < 0.05. Supplementation of PN with 20% EN preserves gut barrier function, by way of maintaining innate immunity, IAP and intestinal microbiota.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-05-01

    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.

  1. Determination of alkaline phosphatase based on affinity adsorption solid-substrate room temperature phosphorimetry using rhodamine 6G-dibromoluciferin luminescent nanoparticle to label lectin and prediction of diseases.

    Science.gov (United States)

    Liu, Jia-Ming; Liu, Zhen-Bo; Hu, Li-Xiang; He, Hang-Xia; Yang, Min-Lan; Zhou, Ping; Chen, Xin-Hua; Zheng, Min-Min; Zeng, Xiao-Yi; Xu, Yue-Long

    2006-10-15

    In the presence of heavy atom perturber LiAc, the silicon dioxide nanoparticle containing rhodamine 6G (R) and dibromoluciferin (D) (R-D-SiO(2)) can emit strong and stable solid-substrate room temperature phosphorescence signal of R (lambda(ex)/lambda(em)=481/648 nm) and D (lambda(ex)/lambda(em)=457/622 nm) on the surface of acetyl cellulose membrane (ACM). R-D-SiO(2) is used to label triticum vulgare lectin (WGA). Then two types of affinity adsorption reactions, R-D-SiO(2)-WGA- alkaline phosphatase (ALP) (direct method) and WGA-ALP-WGA-R-D-SiO(2) (sandwich method), are carried out on ACM. The conditions and the analytical characteristics for the determination of ALP using affinity adsorption solid-substrate room temperature phosphorimetry (AA-SS-RTP) were studied. For a 0.40-microl drop of sample, results show that the detection limits of the sandwich method are 0.16 ag spot(-1)(457/622 nm) and 0.17 ag spot(-1)(481/648 nm), and the detection limits of the direct method are 0.41 ag spot(-1) (457/622 nm) and 0.44 ag spot(-1) (481/648 nm). The contents of ALP in human serum correlated well with those obtained by enzyme-linked immunoassay. This study shows that AA-SS-RTP whether by the sandwich method or the direct method, can combine very well the characteristics of both high sensitivity of SS-RTP and specificity of the immunoreaction. Simultaneously, whether the phosphorescence excitation/emission wavelength of either R or D in R-D-SiO(2) is chosen to determine ALP, this can promote the agility and widen the adaptability of AA-SS-RTP. PMID:16942743

  2. Immunocytochemical Phenotyping of Disseminated Tumor Cells in Bone Marrow by uPA Receptor and CK18: Investigation of Sensitivity and Specificity of an Immunogold/Alkaline Phosphatase Double Staining Protocol

    OpenAIRE

    Allgayer, Heike; Heiss, Markus Maria; Riesenberg, Rainer; Babic, Rudolf; Jauch, Karl Walter; Schildberg, Friedrich Wilhelm

    1997-01-01

    Phenotyping of cytokeratin (CK) 18-positive cells in bone marrow is gaining increasing importance for future prognostic screening of carcinoma patients. Urokinase-type plasminogen activator receptor (uPA-R) is one example of a potential aggressive marker for those cells. However, a valid and reliable double staining method is needed. Using monoclonal antibodies against uPA-R and CK18, we modified an immunogold/alkaline phosphatase double staining protocol. UPA-R/CK18-positive tumor cell contr...

  3. 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)

    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

    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.

  4. Overexpression of tissue-nonspecific alkaline phosphatase increases the expression of neurogenic differentiation markers in the human SH-SY5Y neuroblastoma cell line.

    Science.gov (United States)

    Graser, Stephanie; Mentrup, Birgit; Schneider, Doris; Klein-Hitpass, Ludger; Jakob, Franz; Hofmann, Christine

    2015-10-01

    Patients suffering from the rare hereditary disease hypophosphatasia (HPP), which is based on mutations in the ALPL gene, tend to develop central nervous system (CNS) related issues like epileptic seizures and neuropsychiatric illnesses such as anxiety and depression, in addition to well-known problems with the mineralization of bones and teeth. Analyses of the molecular role of tissue-nonspecific alkaline phosphatase (TNAP) in transgenic SH-SY5Y(TNAPhigh) neuroblastoma cells compared to SH-SY5Y(TNAPlow) cells indicate that the enzyme influences the expression levels of neuronal marker genes like RNA-binding protein, fox-1 homolog 3 (NEUN) and enolase 2, gamma neuronal (NSE) as well as microtubule-binding proteins like microtubule-associated protein 2 (MAP2) and microtubule-associated protein tau (TAU) during neurogenic differentiation. Fluorescence staining of SH-SY5Y(TNAPhigh) cells reveals TNAP localization throughout the whole length of the developed projection network and even synapsin ? co-localization with strong TNAP signals at some spots at least at the early time points of differentiation. Additional immunocytochemical staining shows higher MAP2 expression in SH-SY5Y(TNAPhigh) cells and further a distinct up-regulation of tau and MAP2 in the course of neurogenic differentiation. Interestingly, transgenic SH-SY5Y(TNAPhigh) cells are able to develop longer cellular processes compared to control cells after stimulation with all-trans retinoic acid (RA). Current therapies for HPP prioritize improvement of the bone phenotype. Unraveling the molecular role of TNAP in extraosseous tissues, like in the CNS, will help to improve treatment strategies for HPP patients. Taking this rare disease as a model may also help to dissect TNAP's role in neurodegenerative diseases and even improve future treatment of common pathologies. PMID:26032516

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

    Directory of Open Access Journals (Sweden)

    Marta D. Van Loan

    2012-02-01

    Full Text Available In spite of a high prevalence of vitamin D inadequacy in pregnant women and neonates, relationships among vitamin D status (25(OHD, parathyroid hormone (PTH, bone specific alkaline phosphatase (BALP, and whole body bone mineral content (WBBMC in the newborn are poorly characterized. The purpose of the present study was to investigate the relationships between maternal and cord 25(OHD, PTH, BALP, and WBBMC in newborns in a multiethnic population in Oakland, California and to evaluate the predictive value of the biochemical indices as indicators of WBBMC. Maternal and cord blood were collected from 80 mother-infant pairs and infant WBBMC was measured by dual energy X-ray absorptiometry 8–21 days post-birth. Cord PTH and BALP were each inversely correlated with infant WBBMC (r = ?0.28, p = 0.01 and r = ?0.26, p = 0.02 and with cord 25(OHD (r = ?0.24, p = 0.03 and r = ?0.34, p = 0.002, while cord 25(OHD and unadjusted or weight-adjusted WBBMC were not significantly correlated with one other. In multivariate regression modeling, infant WBBMC was most strongly predicted by infant weight ( p < 0.0001, while either PTH or BALP contributed modestly but significantly to the model (p = 0.006 and p = 0.03 respectively. Cord 25(OHD was not a significant predictor of infant WBBMC. This study provides evidence of associations between feto-maternal 25(OHD, cord PTH and BALP, and early infant WBBMC, though neither feto-maternal 25(OHD nor the measured biochemical indices were suitable indicators of WBBMC.

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

    Science.gov (United States)

    Yang, Yunlong; Zhu, Yu Cheng; Ottea, James; Husseneder, Claudia; Leonard, B Rogers; Abel, Craig; Luttrell, Randall; Huang, Fangneng

    2011-01-01

    The sugarcane borer, Diatraea saccharalis, is a major target pest of transgenic corn expressing Bacillus thuringiensis (Bt) proteins (i.e., Cry1Ab) in South America and the mid-southern region of the United States. Evolution of insecticide resistance in such target pests is a major threat to the durability of transgenic Bt crops. Understanding the pests' resistance mechanisms will facilitate development of effective strategies for delaying or countering resistance. Alterations in expression of cadherin- and alkaline phosphatase (ALP) have been associated with Bt resistance in several species of pest insects. In this study, neither the activity nor gene regulation of ALP was associated with Cry1Ab resistance in D. saccharalis. Total ALP enzymatic activity was similar between Cry1Ab-susceptible (Cry1Ab-SS) and -resistant (Cry1Ab-RR) strains of D. saccharalis. In addition, expression levels of three ALP genes were also similar between Cry1Ab-SS and -RR, and cDNA sequences did not differ between susceptible and resistant larvae. In contrast, altered expression of a midgut cadherin (DsCAD1) was associated with the Cry1Ab resistance. Whereas cDNA sequences of DsCAD1 were identical between the two strains, the transcript abundance of DsCAD1 was significantly lower in Cry1Ab-RR. To verify the involvement of DsCAD1 in susceptibility to Cry1Ab, RNA interference (RNAi) was employed to knock-down DsCAD1 expression in the susceptible larvae. Down-regulation of DsCAD1 expression by RNAi was functionally correlated with a decrease in Cry1Ab susceptibility. These results suggest that down-regulation of DsCAD1 is associated with resistance to Cry1Ab in D. saccharalis. PMID:21991350

  7. Genetic toggling of alkaline phosphatase folding reveals signal peptides for all major modes of transport across the inner membrane of bacteria.

    Science.gov (United States)

    Marrichi, Matthew; Camacho, Luis; Russell, David G; DeLisa, Matthew P

    2008-12-12

    Prediction of export pathway specificity in prokaryotes is a challenging endeavor due to the similar overall architecture of N-terminal signal peptides for the Sec-, SRP- (signal recognition particle), and Tat (twin arginine translocation)-dependent pathways. Thus, we sought to create a facile experimental strategy for unbiased discovery of pathway specificity conferred by N-terminal signals. Using a limited collection of Escherichia coli strains that allow protein oxidation in the cytoplasm or, conversely, disable protein oxidation in the periplasm, we were able to discriminate the specific mode of export for PhoA (alkaline phosphatase) fusions to signal peptides for all of the major modes of transport across the inner membrane (Sec, SRP, or Tat). Based on these findings, we developed a mini-Tn5 phoA approach to isolate pathway-specific export signals from libraries of random fusions between exported proteins and the phoA gene. Interestingly, we observed that reduced PhoA was exported in a Tat-independent manner when targeted for Tat export in the absence of the essential translocon component TatC. This suggests that initial docking to TatC serves as a key specificity determinant for Tat-specific routing of PhoA, and in its absence, substrates can be rerouted to the Sec pathway, provided they remain compatible with the Sec export mechanism. Finally, the utility of our approach was demonstrated by experimental verification that four secreted proteins from Mycobacterium tuberculosis carrying putative Tat signals are bona fide Tat substrates and thus represent potential Tat-dependent virulence factors in this important human pathogen. PMID:18819916

  8. Short-term moderate hypothermia stimulates alkaline phosphatase activity and osteocalcin expression in osteoblasts by upregulating Runx2 and osterix in vitro.

    Science.gov (United States)

    Aisha, M D; Nor-Ashikin, M N K; Sharaniza, A B; Nawawi, H M; Kapitonova, M Y; Froemming, G R A

    2014-08-01

    Exposure of Normal Human Osteoblast cells (NHOst) to a period of hypothermia may interrupt their cellular functions, lead to changes in bone matrix and disrupt the balance between bone formation and resorption, resulting in bone loss or delayed fracture healing. To investigate this possibility, we exposed NHOst cells to moderate (35 °C) and severe (27 °C) hypothermia for 1, 12, 24 and 72 h. The effects of hypothermia with respect to cell cytoskeleton organization, metabolic activity and the expression of cold shock chaperone proteins, osteoblast transcription factors and functional markers, were examined. Our findings showed that prolonged moderate hypothermia retained the polymerization of the cytoskeletal components. NHOst cell metabolism was affected differently according to hypothermia severity. The osteoblast transcription factors Runx2 and osterix were necessary for the transcription and translation of bone matrix proteins, where alkaline phosphatase (Alp) activity and osteocalcin (OCN) bone protein were over expressed under hypothermic conditions. Consequently, bone mineralization was stimulated after exposure to moderate hypothermia for 1 week, indicating bone function was not impaired. The cold shock chaperone protein Rbm3 was significantly upregulated (p<0.001) during the cellular stress adaption under hypothermic conditions. We suggest that Rbm3 has a dual function: one as a chaperone protein that stabilizes mRNA transcripts and a second one in enhancing the transcription of Alp and Ocn genes. Our studies demonstrated that hypothermia permitted the in vitro maturation of NHOst cells probably through an osterix-dependent pathway. For that reason, we suggest that moderate hypothermia can be clinically applied to counteract heat production at the fracture site that delays fracture healing. PMID:24928274

  9. 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.

    Science.gov (United States)

    Dong, Jie-Xian; Li, Zhen-Feng; Lei, Hong-Tao; Sun, Yuan-Ming; Ducancel, Frédéric; Xu, Zhen-Lin; Boulain, Jean-Claude; Yang, Jin-Yi; Shen, Yu-Dong; Wang, Hong

    2012-07-29

    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(H) and V(L)) from hybridoma cell line AC2, which secretes antibodies against RAC, and assembling V(H) and V(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(-1), respectively, and the linear response range extended from 0.05 to 1.45 ng mL(-1). The assay was 10 times as sensitive as the corresponding enzyme-linked immunosorbent assay based on the same fusion protein. Cross-reactivity studies showed that the fusion protein did not cross react with RAC analogs. DcCLEIA was used to analyze RAC spiked pork samples, and the validation was confirmed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS). The results showed a good correlation between the data of dc-CLEIA and HPLC-MS (R(2)>0.99), indicating that the assay was an efficient analytical method for monitoring food safety. PMID:22769009

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

    Directory of Open Access Journals (Sweden)

    C. Sousa

    2011-08-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Liu Hsin-Yi

    2010-04-01

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

  12. 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)

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

    2005-10-01

    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.

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

    International Nuclear Information System (INIS)

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

  14. 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)

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

    2012-11-01

    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

  15. 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)

    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

    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.

  16. Differential Role of Manduca sexta Aminopeptidase-N and Alkaline Phosphatase in the Mode of Action of Cry1Aa, Cry1Ab, and Cry1Ac Toxins from Bacillus thuringiensis

    OpenAIRE

    Flores-Escobar, Biviana; Rodríguez-Magadan, Hector; Bravo, Alejandra; Soberón, Mario; Gómez, Isabel

    2013-01-01

    Aminopeptidase-N (APN1) and alkaline phosphatase (ALP) proteins located in the midgut epithelium of Manduca sexta have been implicated as receptors for Cry1Aa, Cry1Ab, and Cry1Ac insecticidal proteins produced by Bacillus thuringiensis subsp. kurstaki. In this study, we analyzed the roles of ALP and APN1 in the toxicity of these three Cry1A proteins. Ligand blot analysis using brush border membrane vesicles of M. sexta showed that Cry1Aa and Cry1Ab bind preferentially to ALP during early inst...

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

    OpenAIRE

    1992-01-01

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

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

    OpenAIRE

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

    2015-01-01

    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 ...

  19. Structure of Acid phosphatases.

    Science.gov (United States)

    Araujo, César L; Vihko, Pirkko T

    2013-01-01

    Acid phosphatases are enzymes that have been studied extensively due to the fact that their dysregulation is associated with pathophysiological conditions. This characteristic has been exploited for the development of diagnostic and therapeutic methods. As an example, prostatic acid phosphatase was the first marker for metastatic prostate cancer diagnosis and the dysregulation of tartrate resistant acid phosphatase is associated with abnormal bone resorption linked to osteoporosis. The pioneering crystallization studies on prostatic acid phosphatase and mammalian tartrate-resistant acid phosphatase conformed significant milestones towards the elucidation of the mechanisms followed by these enzymes (Schneider et al., EMBO J 12:2609-2615, 1993). Acid phosphatases are also found in nonmammalian species such as bacteria, fungi, parasites, and plants, and most of them share structural similarities with mammalian acid phosphatase enzymes. Acid phosphatase (EC 3.1.3.2) enzymes catalyze the hydrolysis of phosphate monoesters following the general equation. Phosphate monoester + H2O -->/kinds of phosphatase activity. The search for more sensitive and specific methods of detection in clinical laboratory applications led to the development of radioimmunoassays (RIA) for determination of prostatic acid phosphatase in serum. These methods permit the direct quantification of the enzyme regardless of its activity status. Therefore, an independent structural classification exists that helps to group these enzymes according to their structural features and mechanisms. Based on this we can distinguish the histidine acid phosphatases (Van Etten, Ann N Y Acad Sci 390:27-51, 1982), the low molecular weight protein tyrosine acid phosphatases and the metal-ion dependent phosphatases. A note of caution is worthwhile mentioning here. The nomenclature of acid phosphatases has not been particularly easy for those new to the subject. Unfortunately, the acronym PAP is very common in the literature about purple acid phosphatases and prostatic acid phosphatase. In addition, LPAP is the acronym chosen to refer to the lysophosphatidic acid phosphatase which is a different enzyme. It is important to bear in mind this distinction while reviewing the literature to avoid confusion. PMID:23860654

  20. 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

    Directory of Open Access Journals (Sweden)

    V. Bertone

    2011-02-01

    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.

  1. 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)

    Jean-Paul, Lallès; Jaime Parra, Suescún.

    2014-06-01

    Full Text Available 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. Various intestinal diseases, including necrotizing enterocolit

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

    Directory of Open Access Journals (Sweden)

    M.A. Geidam

    2007-01-01

    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.

  3. Prostatic acid phosphatase in serum and semen of dogs / Fosfatasa ácida prostática en suero y semen de perros

    Scientific Electronic Library Online (English)

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

    Full Text Available La incidencia de cáncer de próstata ha incrementado el uso de los marcadores celulares para detectar el cáncer en este tejido. Antígenos específicos del tejido o antígenos de diferenciación se encuentran en la superficie de las células normales. Clínicamente, estos antígenos son importantes para el [...] diagnóstico de alteraciones en estos tejidos y para la inmunoterapia. Este estudio trata de evaluar la importancia de la fosfatasa ácida prostática en la próstata canina e investigar su concentración en el suero y en el plasma seminal de perros saludables de diferentes edades. La concentración de fosfatasa ácida prostática en el plasma seminal y en el suero fue evaluada por espectrofotometría, utilizando un kit comercial. Los niveles de la fosfatasa ácida prostática (PAP) no fueron diferentes de acuerdo con la edad y no presentaron correlación con la edad o con las dimensiones de las próstatas verificadas por ecografía. Los valores de concentración de PAP presentaron una gran variación en cada grupo. Sin embargo, son necesarios más estudios para evaluar el papel de la fosfatasa ácida prostática en la próstata canina y su importancia como una prueba de diagnóstico para los trastornos de la próstata. Abstract in english The incidence of prostatic malignancy has increased the use of tissue markers to detect cancer. Tissue specific antigens or differentiation antigens are found on the surface of normal cells. Clinically, these antigens are important to diagnose alterations in the tissues and for immunotherapy. The ob [...] jective of the present study was to evaluate the prostatic acid phosphatase concentration in blood and seminal plasma of intact and healthy dogs at different ages. The evaluation was carried out by spectrophotometer, using a commercial kit. The prostatic acid phosphatase (PAP) levels did not differ according to the age and did not correlate with age or prostatic dimensions verified by ultrasonography. The PAP concentration values varied greatly within each group. However, more studies are necessary to evaluate the role of prostatic acid phosphatase in the canine prostate and its importance as a diagnostic test for prostate disorders.

  4. 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...

  5. 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)

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

    2013-08-01

    Full Text Available 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.

  6. Experimental study on the usefulness of magnetotherapy in bone fractures (tibial osteotomy in the rat). Accumulation of 99 mTc MDP - tests of tensile strength - determination of alkaline phosphatase

    International Nuclear Information System (INIS)

    Non-directional magnetic field therapy using a flux density of 60 G and a frequency of 25 Hz was carried out over 12 hours daily in rats in order to ascertain its influence on the healing process following osteotomy of the tibia with internal splint fixation of the fractured bone being carried out as an additional measure. The results thus achieved were compared to those seen in control animals, were no magnetotherapy was carried out, on the basis of scintiscan studies using 99 mTc MDP (degree of density in the callus formed around the fracture zone), the plasma levels of alkaline phosphatase and tests of tensile strength. The follow-up observations of the healing process were additionally based on radiological and histological evaluations of the animals. Beneficial effects of magnetotherapy on the healing process could not be confirmed with any statistical significance. (TRV)

  7. 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)

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

    2003-06-01

    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.

  8. Hematology and serum chemistry of cottontail rabbits of southern Illinois.

    Science.gov (United States)

    Lepitzki, D A; Woolf, A

    1991-10-01

    In 1983 and 1984 blood was collected from 79 cottontail rabbits (Sylvilagus floridanus) confined to an outdoor enclosure in southern Illinois to establish reference values for hematology and serum chemistry. Packed cell volume, sodium, potassium, chloride, glucose, calcium, carbon dioxide, blood urea nitrogen, creatinine, uric acid, cholesterol, albumin, bilirubin, alkaline phosphatase, aspartate transaminase, alanine aminotransaminase, total protein, albumin/globulin ratio, and osmolality were measured. Sex and age (adult versus juvenile) of rabbit as well as season (June to September versus October to May) and method of capture (trap versus shot) variously affected most hematology and serum chemistry variables. PMID:1758030

  9. Differential role of Manduca sexta aminopeptidase-N and alkaline phosphatase in the mode of action of Cry1Aa, Cry1Ab, and Cry1Ac toxins from Bacillus thuringiensis.

    Science.gov (United States)

    Flores-Escobar, Biviana; Rodríguez-Magadan, Hector; Bravo, Alejandra; Soberón, Mario; Gómez, Isabel

    2013-08-01

    Aminopeptidase-N (APN1) and alkaline phosphatase (ALP) proteins located in the midgut epithelium of Manduca sexta have been implicated as receptors for Cry1Aa, Cry1Ab, and Cry1Ac insecticidal proteins produced by Bacillus thuringiensis subsp. kurstaki. In this study, we analyzed the roles of ALP and APN1 in the toxicity of these three Cry1A proteins. Ligand blot analysis using brush border membrane vesicles of M. sexta showed that Cry1Aa and Cry1Ab bind preferentially to ALP during early instars while binding to APN was observed after the third instar of larval development. Cry1Ac binds to APN throughout all larval development, with no apparent binding to ALP. ALP was cloned from M. sexta midgut RNA and expressed in Escherichia coli. Surface plasmon resonance binding analysis showed that recombinant ALP binds to Cry1Ac with 16-fold lower affinity than to Cry1Aa or Cry1Ab. Downregulation of APN1 and ALP expression by RNA interference (RNAi) using specific double-stranded RNA correlated with a reduction of transcript and protein levels. Toxicity analysis of the three Cry1A proteins in ALP- or APN1-silenced larvae showed that Cry1Aa relies similarly on both receptor molecules for toxicity. In contrast, RNAi experiments showed that ALP is more important than APN for Cry1Ab toxicity, while Cry1Ac relied principally on APN1. These results indicated that ALP and APN1 have a differential role in the mode of action of Cry1A toxins, suggesting that B. thuringiensis subsp. kurstaki produces different Cry1A toxins that in conjunction target diverse midgut proteins to exert their insecticidal effect. PMID:23686267

  10. The Intestinal Epithelial Cell Differentiation Marker Intestinal Alkaline Phosphatase (ALPi) Is Selectively Induced by Histone Deacetylase Inhibitors (HDACi) in Colon Cancer Cells in a Kruppel-like Factor 5 (KLF5)-dependent Manner*

    Science.gov (United States)

    Shin, Joongho; Carr, Azadeh; Corner, Georgia A.; Tögel, Lars; Dávaos-Salas, Mercedes; Tran, Hoanh; Chueh, Anderly C.; Al-Obaidi, Sheren; Chionh, Fiona; Ahmed, Naseem; Buchanan, Daniel D.; Young, Joanne P.; Malo, Madhu S.; Hodin, Richard A.; Arango, Diego; Sieber, Oliver M.; Augenlicht, Leonard H.; Dhillon, Amardeep S.; Weber, Thomas K.; Mariadason, John M.

    2014-01-01

    The histone deacetylase inhibitor (HDACi) sodium butyrate promotes differentiation of colon cancer cells as evidenced by induced expression and enzyme activity of the differentiation marker intestinal alkaline phosphatase (ALPi). Screening of a panel of 33 colon cancer cell lines identified cell lines sensitive (42%) and resistant (58%) to butyrate induction of ALP activity. This differential sensitivity was similarly evident following treatment with the structurally distinct HDACi, MS-275. Resistant cell lines were significantly enriched for those harboring the CpG island methylator phenotype (p = 0.036, Chi square test), and resistant cell lines harbored methylation of the ALPi promoter, particularly of a CpG site within a critical KLF/Sp regulatory element required for butyrate induction of ALPi promoter activity. However, butyrate induction of an exogenous ALPi promoter-reporter paralleled up-regulation of endogenous ALPi expression across the cell lines, suggesting the presence or absence of a key transcriptional regulator is the major determinant of ALPi induction. Through microarray profiling of sensitive and resistant cell lines, we identified KLF5 to be both basally more highly expressed as well as preferentially induced by butyrate in sensitive cell lines. KLF5 overexpression induced ALPi promoter-reporter activity in resistant cell lines, KLF5 knockdown attenuated butyrate induction of ALPi expression in sensitive lines, and butyrate selectively enhanced KLF5 binding to the ALPi promoter in sensitive cells. These findings demonstrate that butyrate induction of the cell differentiation marker ALPi is mediated through KLF5 and identifies subsets of colon cancer cell lines responsive and refractory to this effect. PMID:25037223

  11. 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

    Directory of Open Access Journals (Sweden)

    Berta B Socarrás Ferrer

    2001-04-01

    Full Text Available 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 mieloproliferativosThe 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

  12. 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)

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

    2001-04-01

    Full Text Available 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

  13. 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)

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

    2013-12-01

    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.

  14. 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

  15. 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)

    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

    Full Text Available 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.

  16. 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

  17. Passive immunity transfer and serum constituents of crossbred calves

    Directory of Open Access Journals (Sweden)

    Thaís G. Rocha

    2012-06-01

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

  18. 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

  19. 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.)

  20. Study on the Changes in Enzyme and Insulin-like Growth Factor-1 Concentrations in Blood Serum and Growth Characteristics of Velvet Antler during the Antler Growth Period in Sika Deer (Cervus nippon).

    Science.gov (United States)

    Park, Jaehyun; Jeon, Byongtae; Kang, Sungki; Oh, Mirae; Kim, Myonghwa; Jang, Seyoung; Park, Pyojam; Kim, Sangwoo; Moon, Sangho

    2015-09-01

    This study was conducted to investigate changes in blood enzyme parameters and to evaluate the relationship between insulin-like growth factor-1 (IGF-1), antler growth and body weight during the antler growth of sika deer (Cervus nippon). Serum enzyme activity and IGF-1 concentrations were measured in blood samples collected from the jugular and femoral veins at regular intervals during the antler growth period. Blood samples were taken in the morning from fasted stags (n = 12) which were healthy and showed no clinical signs of disease. Alfalfa was available ad libitum and concentrates were given at 1% of body weight to all stags. The experimental diet was provided at 9 am with water available at all times. There were no significant differences in alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase during antler growth, but alkaline phosphatase concentrations increased with antler growth progression, and the highest alkaline phosphatase concentration was obtained 55 days after antler casting. Serum IGF-1 concentrations measured from blood samples taken from the jugular vein during antler growth, determined that levels of IGF-1 was associated with body weight and antler growth patterns. Serum IGF-1 concentrations were higher at the antler cutting date than other sampling dates. Antler length increased significantly during antler growth (palkaline phosphatase concentration was related to antler growth and both antler growth and body weight were associated positively with IGF-1 concentrations during antler growth. PMID:26194228

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

    Directory of Open Access Journals (Sweden)

    N.E. Abdelmeguid

    2000-01-01

    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.

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

    OpenAIRE

    von Tigerstrom, R G

    1984-01-01

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

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

    Directory of Open Access Journals (Sweden)

    CORRYANTI

    2007-07-01

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

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

    Scientific Electronic Library Online (English)

    N.J.F., Oliveira; M.M., Melo; L.A., Lago; E.F., Nascimento.

    2005-06-01

    Full Text Available [...] Abstract in english Hemogram and serum biochemistry (aspartate aminotransferase, alkaline phosphatase and gamma glutamiltransferase, total protein, urea, creatinine, calcium and phosphorus) were performed weekly in five crossbreed bovine after consumption of a diet containing citrus pulp pellets (40%), for 43 days. Per [...] cutaneous hepatic biopsy and histologic evaluation were performed in each animal before and after consumption of the citrus pulp diet. Hemogram, the enzymes aspartate aminotransferase and gamma glutamiltransferase, urea and creatinine had normal levels at the end of the experiment. No histologic lesions were observed in liver samples before or after citrus pulp consumption. However, there was an increase of serum phosphorus and reduction of serum calcium (p

  5. 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)

  6. Oral toxic exposure of titanium dioxide nanoparticles on serum biochemical changes in adult male Wistar rats

    Directory of Open Access Journals (Sweden)

    Dasal Vasantharaja

    2015-01-01

    Full Text Available Objective(s: Titanium dioxide (TiO2 nanoparticles (NPs are widely used in commercial food additives and cosmetics worldwide. Uptake of these nanoparticulate into humans by different routes and may exhibit potential side effects, lags behind the rapid development of nanotechnology. Thus, the present study designed to evaluate the toxic effect of mixed rutile and anatase TiO2 NPs on serum biochemical changes in rats. Materials and Methods: In this study, adult male Wistar rats were randomly allotted into the experimental and control groups (n=6, which were orally administered with 50 and 100 mg/kg body weight of TiO2 NPs. Toxic effects were assessed by the changes of serum biochemical parameters such as glucose, total protein, albumin, globulin, cholesterol, triglyceride, high density lipoprotein, alanine transaminase, aspartate transaminase, alkaline phosphatase, total bilirubin, blood urea nitrogen, uric acid and creatinine. All the serum biochemical markers were experimented in rats, after 14-days of post exposure. Results: Changes of the serum specific parameters indicated that liver and kidney were significantly affected in both experimental groups. The changes between the levels of total protein, glucose, aspartate transaminase, alanine transaminase and alkaline phosphatase indicate that TiO2 NPs induces liver damage. Significant increase in the blood urea nitrogen and uric acid indicates the renal damage in the TiO2 NPs treated rats. Conclusion: The data shows that the oral administration of TiO2 NPs (

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

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    Babi? Olivera B.

    2013-01-01

    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

  8. 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)

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

    2011-02-01

    Full Text Available 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

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

    OpenAIRE

    N. J. F. Oliveira; M.M. Melo; L. A. Lago; E.F Nascimento

    2005-01-01

    Hemogram and serum biochemistry (aspartate aminotransferase, alkaline phosphatase and gamma glutamiltransferase, total protein, urea, creatinine, calcium and phosphorus) were performed weekly in five crossbreed bovine after consumption of a diet containing citrus pulp pellets (40%), for 43 days. Percutaneous hepatic biopsy and histologic evaluation were performed in each animal before and after consumption of the citrus pulp diet. Hemogram, the enzymes aspartate aminotransferase and gamma glu...

  10. The attachment of serum- and plasma-derived C3 to solid-phase immune aggregates and its relation to complement-mediated solubilization of immune complexes

    DEFF Research Database (Denmark)

    Baatrup, G; Svehag, S E; Jensenius, J C

    1986-01-01

    The interaction between immune aggregates and complement (C) was investigated. Solid-phase immune aggregates were prepared by coating microwells with heat-aggregated bovine serum albumin (BSA) followed by rabbit anti-BSA antibody. The immune aggregates were reacted with human serum or citrated plasma at 37 degrees C. The binding of C3 components was investigated with biotinylated F(ab')2 antibodies to C3c and C3d and avidin-coupled alkaline phosphatase. The form of the incorporated C3, whether C...

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

    International Nuclear Information System (INIS)

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

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

    OpenAIRE

    Maria-José BOAVIDA; Geraldes, Ana Maria

    2003-01-01

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

  13. 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

    Directory of Open Access Journals (Sweden)

    Sérgio R. Nozawa

    2002-01-01

    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.

  14. Extremely low serum pyridoxal 5'-phosphate in children with familial hypophosphatemic rickets.

    Science.gov (United States)

    Reynolds, R D; Lorenc, R S; Wieczorek, E; Pronicka, E

    1991-03-01

    The apparent vitamin B-6 status of 31 children with familial hypophosphatemic rickets (FHR) was determined. All children had alkaline phosphatase activity that was high-normal to elevated for their ages. A sensitive assay for pyridoxal 5'-phosphate (PLP) indicated that 15 of the 31 children had an undetectable (less than 0.2 nmol/L) concentration of the vitamer--the lowest values yet reported in human serum. The 16 remaining children had concentrations of the vitamer so low that they indicated a potential severe vitamin B-6 deficiency. However, none of the children had ever presented with any of the classical vitamin B-6-deficiency symptoms. Treatment of three additional FHR children with 100 mg pyridoxine.HCl/d resulted in a moderate and transient elevation of their serum PLP concentrations, a dramatic elevation of their erythrocyte PLP concentrations, and no improvement in clinical condition. Serum or plasma PLP concentrations are an inappropriate index for determining vitamin B-6 status in people with FHR and perhaps in others with elevated alkaline phosphatase activity. PMID:2000825

  15. Sensitivity and specificity of serum cross-linked N-telopeptides of type 1 collagen (NTx in patients with solid tumors

    Directory of Open Access Journals (Sweden)

    Fernando Jablonka

    2009-03-01

    Full Text Available Objective: Cross-linked N-telopeptides of type I collagen (NTx increase in situations in which bone resorption is increased, such as osteoporosis and bone metastasis. It was evaluated the usefulness of NTx for the diagnosis of bone metastases in patients with solid tumors. Methods: It was conducted a prospective study, at a tertiary public hospital. A quantitative enzyme-linked immunosorbent assay (ELISA assay of serum NTx levels in 19 subjects without a history of cancer and 62 patients with various solid tumors referred for a bone scan was evaluated. Three specialists read all bone scans. Rresults: It was not found any significant correlations between serum NTx and age, sex, history of bone pain, tumor type and bone alkaline phosphatase levels. We found a significant correlation between serum NTx and alkaline phosphatase levels (r2 = 0.08, p  = 0.022. Serum NTx sensitivity, specificity, positive and negative predictive values and area under the receiver operating characteristic (ROC curve for the presence of bone metastases were 0.34, 0.92, 0.95, 0.22 and 0.59, respectively. Cconclusions: It was concluded that serum levels of NTx are highly sensitive for the diagnosis of bone metastases in patients with solid tumors.

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

    Directory of Open Access Journals (Sweden)

    PRL Silva

    2007-12-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Sunita Pujar

    2011-10-01

    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

  18. 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. (authors)

  19. Stool vs. Serum Hepatitis B Virus DNA in Patients with Chronic Hepatitis B

    Science.gov (United States)

    Zheng, Ji-shun; Chen, Meng-meng; Yang, Hai-fei; Zhou, Xiang-tian; Liu, Yan-yan; Li, Jia-bin

    2015-01-01

    Background Serum hepatitis B virus (HBV) DNA and hepatitis B e antigen (HBeAg) liver function in patients with chronic hepatitis B (CHB) are significantly associated. A comparison of clinical significance of fecal HBV DNA and serum HBV DNA has not yet been reported. Material/Methods Stool and serum samples were collected from 66 patients with CHB. Fecal HBV DNA, serum HBV DNA, and intestinal microbiota DNA were detected by real-time quantitative fluorescence polymerase chain reaction (PCR). Liver function and HBeAg were analyzed. Results The stool and serum HBV DNA were positively correlated (r=0.57, P=0.001). Fecal HBV DNA was higher in the HBeAg-positive group than in the HBeAg-negative group (P=0.02). Fecal HBV DNA was negatively correlated with alkaline phosphatase (ALP) (r=?0.41, P=0.001) and TBIL (r=?0.29, P=0.02), and was positively correlated with Enterococcus (r=0.38, P=0.002). Serum HBV DNA was negatively correlated with alanine aminotransferase (ALT) (r=?0.30, P=0.02), aminotransferase (AST) (r=?0.26, P=0.049), and Lactobacillus (r=?0.31, P=0.01). Conclusions These observations suggest that fecal HBV DNA and serum HBV DNA in patients with CHB have different effects. Fecal HBV DNA might be associated with changes in Enterococcus concentrations, but serum HBV DNA is not. PMID:26645150

  20. 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...

  1. Efecto de concentraciones crecientes de Fósforo Fítico sobre la actividad de las enzimas fitasa y fosfatasa alcalina en el epitelio intestinal de ovinos jóvenes. / Effect of Incresing Concentrations of Phytic Phosphorus on the Activity of the Phytase and Alkaline Phosphatase Enzymes in the Intestinal Epithelium of Young Sheep.

    Scientific Electronic Library Online (English)

    Pablo, Pizzani; Susmira, Godoy; Milagro, León; Emma, Rueda; María Virginia, Castañeda; Adelis, Arias.

    2008-02-01

    Full Text Available Con el objetivo de conocer el efecto del suministro de concentraciones crecientes de fósforo fítico sobre la actividad de las enzimas fitasa y fosfatasa alcalina en el epitelio intestinal de ovinos jóvenes, se realizó un experimento con concentraciones crecientes de fósforo fítico: 0% (T1), 40% (T2) [...] , 60% (T3) y 80% (T4). Los animales consumieron las dietas a razón del 4% del peso vivo durante cuatro semanas. Transcurrido este período, se sacrificaron todos los animales del ensayo, y se determinó la actividad de las enzimas en homogenados del epitelio de las diferentes secciones del intestino delgado (duodeno, yeyuno e ileon). Los resultados muestran valores de actividad fitásica a nivel del duodeno de: 1,43; 1,85; 1,82 y 0,69 (nmoles/min/mg de proteína) para concentraciones de T1, T2, T3 y T4, respectivamente, con una disminución significativa (P Abstract in english With the objective to know the effect of supply in growing concentrations of phytic phosphorus on the activity of the enzymes phytase and alkaline phosphatase in the intestinal epithelium of young sheep, it was carried out an experiment with incresing concentration of phytic phosphorus: 0% (T1), 40% [...] (T2), 60% (T3), and 80% (T4). The animals consumed the diets as for 4% of the body weight during four weeks. Lapsed this period, all the animals of the rehearsal were sacrificed, and the activity of the enzymes was determined in homogenates of the epithelium of the different sections of the thin intestine (duodenum, jejunum and ileum). The results showed values of phytase activity at level of the duodenum of: 1.43; 1.85; 1.82 and 0.69 nmoles/min/mg protein for concentrations of T1, T2, T3, and T4, respectively, with a significant decrease (P

  2. Retrospective Study of Serum Sclerostin Measurements in Bed Rest Subjects

    Science.gov (United States)

    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

    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 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.

  3. Concentrations of testosterone, luteal hormone and prolactin in the serum as well as comparisons of sensitivity between radioimmunoassays and enzyme assays for the detection of acid prostate phosphatase in the presence of carcinomas of the prostate

    International Nuclear Information System (INIS)

    The relationship between carcinomas of the prostate and the plasma levels of testosterone, luteal hormone and prolactin as well as the possible influence of these neoplasms on the testosterone binding capacity and free testosterone index are investigated for various tumour stages and degrees of histological differentiation, in connection with several forms of local therapy as well as a variety of contrasexual methods. The sensitivity of enzyme assays and radioimmunoassays for the detection of acid prostate phosphatase is evaluated within the framework of this study. (MBL)

  4. 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)

  5. COMPARATIVE EFFICACY OF CLOVE OIL AND 2-PHENOXY ETHANOL ON SERUM BIOCHEMICAL CHANGES AND HISTOLOGICAL STUDIES IN CHANNAPUNCTATUS

    Directory of Open Access Journals (Sweden)

    G Chelladurai

    2013-05-01

    Full Text Available The aim of the present study was to investigate thecomparison of effect of clove oil and 2-phenoxy ethanol for Channapunctatus and using values of serum biochemical profile and histological tissue studies ,to assess the effects of fish exposure to that anaesthetic:A total of 60 C.punctatus of 60.23±5.02g were divided in to five groups (12 fish per each:1st treatment as control (no anaesthetic,2nd 0.2ml/1 2-phenoxy ethanol ,3rd 0.3ml/1 2-phenoxy ethanol ,4th 30mg/1 clove oil and 5th 40mg/1 clove oil.Biochemical blood profile of C.puntatus were taken 15 min and 24 hrs. After Anaesthesia induction ,the factors used to evaluate the serum biochemical profile included the glucose (GLU, total protein (TP, albumin (ALB,total globulin (GLOP, alkaline phosphatase (ALP,serum Glutamic oxaloacetic Transaminase (SGOT, serum glutamic pyruvic transaminase (SGPT, histological tissue examination of tissue like gills ,ten minutes exposure to 2-phenoxy ethanol and clove oilcaused an increase in concentration of glucose and alkaline phosphatase 15 min after Anaesthesiainduction.Histological examination showed.Capillary ectasia of gill filaments immediately after clove oil and 2-phenoxy ethanol,showed  swelling of primary and secondary lamella .Twenty –four hours after anaesthesia ,no ectasia was observed .No histopathological changes were demonstrated in gills:Our result showed that The 2-phenoxy ethanol at 0.20ml/1and clove oil at 30mg/1 concentration may be used as an efficient and safe anaesthetic for Channapunctatus. Key words: Clove oil, 2-Phenoxy ethanol,Channapunctaus,Serum biochemical profile, histological examination of tissues.

  6. Ivermectin resistant and susceptible third-stage larvae of Haemonchus contortus: cholinesterase and phosphatase activities

    OpenAIRE

    Consuelo Giménez-Pardo; Maria Mercedes Martínez-Grueiro; Alicia Gómez-Barrio; Filomena Rodríguez-Caabeiro

    2004-01-01

    Cholinesterase and acid phosphatase (AP), but not alkaline phosphatase activities, were detected in cytosolic and membrane-bound fractions of ivermectin resistant and susceptible Haemonchus contortus infective-stage larvae. Some differences in acetylcholinesterase activity of cytosolic fractions and in the AP activity of these fractions as well as in the response to AP inhibitors by membrane-bound fractions were detected. Data are discussed.

  7. Ivermectin resistant and susceptible third-stage larvae of Haemonchus contortus: cholinesterase and phosphatase activities

    Directory of Open Access Journals (Sweden)

    Consuelo Giménez-Pardo

    2004-03-01

    Full Text Available Cholinesterase and acid phosphatase (AP, but not alkaline phosphatase activities, were detected in cytosolic and membrane-bound fractions of ivermectin resistant and susceptible Haemonchus contortus infective-stage larvae. Some differences in acetylcholinesterase activity of cytosolic fractions and in the AP activity of these fractions as well as in the response to AP inhibitors by membrane-bound fractions were detected. Data are discussed.

  8. Ivermectin resistant and susceptible third-stage larvae of Haemonchus contortus: cholinesterase and phosphatase activities

    Scientific Electronic Library Online (English)

    Consuelo, Giménez-Pardo; Maria Mercedes, Martínez-Grueiro; Alicia, Gómez-Barrio; Filomena, Rodríguez-Caabeiro.

    2004-03-01

    Full Text Available Cholinesterase and acid phosphatase (AP), but not alkaline phosphatase activities, were detected in cytosolic and membrane-bound fractions of ivermectin resistant and susceptible Haemonchus contortus infective-stage larvae. Some differences in acetylcholinesterase activity of cytosolic fractions and [...] in the AP activity of these fractions as well as in the response to AP inhibitors by membrane-bound fractions were detected. Data are discussed.

  9. Isolation and characterization of gingival fibroblasts positive for alkaline phosphatase in patients with chronic periodontitis and drug-induced gingival hyperplasia / Isolamento e caracterização de fibroblastos gengivais positivos para fosfatase alcalina de pacientes portadores de periodontite crônica e hiperplasia gengival medicamentosa

    Scientific Electronic Library Online (English)

    Vagner Rodrigues, Santos; Rafael Tomaz, Gomes; Maurício, Resende; Oslei Paes de, Almeida; Ricardo Della, Colleta.

    Full Text Available OBJETIVO: Investigar a expressão da atividade de fosfatase alcalina (ALP) em fibroblastos gengivais de sujeitos com periodontite crônica (PC) e hiperplasia gengival medicamentosa (HGM) induzida por difenilhidantoína. METODOLOGIA: Os fragmentos gengivais foram obtidos de 13 pacientes (8 mulheres e 5 [...] homens, 22 a 74 anos), sendo 4 fragmentos procedentes de gengiva clinicamente normal (GN), 5 de biópsias de bolsas periodontais com PC e 4 de HGM induzida pela difenilhidantoína. Através de digestão enzimática, suspensões de células gengivais foram preparadas contendo fibroblastos ALP-positivos sem afetar a atividade da enzima ALP. Análises citoquímica e histoquímica foram realizadas. RESULTADOS: Fibroblastos de GN apresentaram fraca positividade para ALP quando comparados com PC e HGM. PC e HGM exibiram níveis elevados e intermediários, respectivamente, de ALP. Observou-se menor proliferação celular na cultura de células em PC e HGM que em GN. Entretanto, a quantidade de células recuperadas das sub-culturas foi similar à quantidade recuperada na cultura inicial para as três linhagens. CONCLUSÕES: A expressão de ALP na PC e na HGM encontra-se aumentada e ocorre baixa proliferação de fibroblastos na PC e na HGM. Isto sugere que a inflamação periodontal e a difenilhidantoína podem influenciar a expressão de ALP e a multiplicação de fibroblastos gengivais humanos. Outros estudos são necessários para melhor avaliar a importância de ALP no desenvolvimento e na progressão de PC e HGM. Abstract in english PURPOSE: To investigate the expression of alkaline phosphatase (ALP) activity in gingival fibroblasts from individuals with chronic periodontitis (CP) and drug-induced gingival hyperplasia (DGH) induced by diphenylhydantoin. METHODS: Gingival fragments were obtained from 13 patients (8 women and 5 m [...] en, from 22 to 74 years of age), with 4 fragments from clinically normal gingiva (NG), 5 from biopsy of periodontal pockets with CP, and 4 from DGH induced by diphenylhydantoin. Using an enzymatic digestion procedure, gingival cell suspensions containing ALP-positive fibroblasts were prepared without affecting ALP activity. Cytochemistry and histochemistry analyses were performed. RESULTS: Fibroblasts from NG presented low levels of ALP when compared to CP and DGH, which showed elevated and intermediate levels of ALP, respectively. Little cell proliferation was observed for fibroblasts from CP and DGH as compared to NG. However, the quantity of cells recovered from the subcultures was similar to the quantity recovered from the initial cell culture for the three sources. CONCLUSION: The expression of ALP is increased in CP and DGH, and fibroblasts in CP and DGH show low proliferation. This suggests that periodontal inflammation and diphenylhydantoin may influence ALP expression and human gingival fibroblast expansion. Other studies are necessary to better assess the importance of ALP in the development and progression of CP and DGH.

  10. Glucose-6-phosphatase deficiency

    OpenAIRE

    Labrune Philippe; Gajdos Vincent; Eberschweiler Pascale; Hubert-Buron Aurélie; Petit François; Vianey-Saban Christine; Boudjemline Alix; Piraud Monique; Froissart Roseline

    2011-01-01

    Abstract Glucose-6-phosphatase deficiency (G6P deficiency), or glycogen storage disease type I (GSDI), is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, betw...

  11. Effect of two different doses of oral cholecalciferol supplementation on serum 25-hydroxy-vitamin D levels in healthy Indian postmenopausal women: A randomized controlled trial

    Directory of Open Access Journals (Sweden)

    Niti Agarwal

    2013-01-01

    Full Text Available Aim: To compare the effect of two different doses (500 and 1000 IU/day of oral vitamin D3 (cholecalciferol on serum 25-hydroxy vitamin D [25(OHD] levels in apparently healthy postmenopausal Indian women. Materials and Methods: Serum 25(OHD, calcium with albumin, phosphorus, and alkaline phosphatase were measured in 92 apparently healthy postmenopausal women. The subjects were randomly assigned to one of the three groups and received supplementation for 3 months each. Each group received 1000 mg calcium carbonate daily while groups B and C received 500 and 1000 IU of cholecalciferol in addition, respectively. The tests were repeated after 3 months. Results: At baseline, 83.7% subjects had vitamin D deficiency (?20 ng/mL. The difference in the percentage change in mean serum 25(OHD levels from baseline in group A (-30.5 ± 5.3%, group B (+8.9 ± 19.7%, and in group C (+97.8 ± 53.3% was statistically significant (P 20 ng/mL was achieved in 4.7% (1/21, 16% (4/25, and 66.67% (12/18 subjects in groups A, B, and C, respectively. No significant change was found in serum calcium, phosphorus, and alkaline phosphatase levels at 3 months in either of the groups from baseline. Conclusions: Standard dose of cholecalciferol available in "calcium tablets" (250 IU per 500 mg calcium carbonate is not adequate for achieving optimum serum 25(OHD levels in Indian postmenopausal women. Higher dose of vitamin D supplementation with 1000 IU/day (500 IU per 500 mg calcium carbonate daily is superior to the standard dose therapy. For achievement of optimum serum 25(OHD levels (>30 ng/mL in Indian postmenopausal women, still higher doses of vitamin D are likely to be required.

  12. Research on Phosphatases of Belladona Leaves and Their Purification (Part 1

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    M. Khorsand

    1956-12-01

    Full Text Available Belladona leaves as well as all other studied leaves contains two distinct phosphatase fractions belonging respectively to types II and IIIi the major parts of these enzymes is extraetible by water. It was not possible to extract the non soluble fraction which is solidly retained by the cellular constituents. Phosphatase II does not differ from other phosphatnses of the same type. Whereas phosphatase III is distinetely different from enzymes of the same type of vegetal or animal origins. It is activated by bivalent metallic ions which are specific activators of the alkaline phcspbatnses: Mg-Zn-Ni and Co.

  13. Pattern of serum vitamin d in opd patients

    International Nuclear Information System (INIS)

    To find out the prevalence of Vitamin-D deficiency in conditions other than osteomalcia and rickets in our part of the world. Only those patients were included who had any structural or biochemical changes in the body. Serum Vitamin-D level of all patients was sent for estimation from a single reputable laboratory, apart from serum calcium, phosphorus, alkaline phosphatase and routine blood investigations. All data was collected and processed on SPSS Version 10. Of the total 79 patients, 58(73%) were females and 21(27%) males. Minimum age was one year and maximum 90 years, with a mean age of 41.91 and standard deviation of 19.1. Majority of the patients were seen in the 4th, 5th and 6th decades of life, and most of them were house wives. The serum Vitamin-D level was found low in 73(92%) patients and the most severe form of deficiency was seen in patients with tuberculosis. Vitamin-D deficiency was seen in 92% of our patients, belonging to all age groups and suffering from different diseases. (author)

  14. Serum enzymes activities in Plasmodium falciparum infection in Southern Pakistan

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    Koay Yen Chin

    2011-05-01

    Full Text Available Objective: Serum levels of lactate dehydrogenase (LDH,aspartate aminotranferase (AST, alanine aminotransferase(ALT, and alkaline phosphatase (ALP were assessed todetermine the liver functions of patients infected withPlasmodium falciparum. The enzyme activities were assessedin 60 malarial patients and a control group of 44 people.Materials and Methods: The data for the study was collectedfrom the survey conducted from Liaquat University of medicaland health sciences Hospital, Hyderabad, Pakaistan. Sample of60 patients aged between 20 and 50 years were collected. Acontrol group of 44 healthy individual adults was also assessedfor comparative purposes. All the malaria patients who visitedthe OPD during the study period enrolled in the study.Results: The LDH activity in male patients was found to be674.89 ± 33.354 IU/L. This is above the control LDH activity of296.59 ± 14.476 IU/L. Similarly, in female patients, the serumLDH activity of 580.25 ± 24.507 IU/L is over twice the controlfemale serum LDH activity of 302.18 ± 18.082 IU/L. Furtherone-way anova test was performed to find any significance ininfected and control male and female.Conclusion: Hepatic dysfunction was found to be associated toP. falciparum malaria infection.

  15. Trends and physiology of common serum biochemistries in children aged 0-18 years.

    Science.gov (United States)

    Loh, Tze Ping; Metz, Michael Patrick

    2015-08-01

    The aim of this study was to visually present and discuss in detail the physiological trends of 22 serum biochemistries in children aged 0-18.A data-mining, LMS (lambda, mu, and sigma) approach was employed to derive the smoothed continuous serum biochemistry centile charts, after application of stringent outlier exclusion criteria.Serum sodium and calculated osmolality are low in early life and rise with age due to maturing kidney and body water redistribution. Urea, creatinine and uric acid is high at birth, declines to reach a trough by 1 month of age and gradually rises again thereafter. Serum bicarbonate falls initially during the neonatal and toddler period, then rises with declining respiratory rate, further increasing sodium and suppressing chloride. Potassium, calcium and phosphate are required for somatic growth and are actively accrued during periods of rapid growth. Albumin increases until puberty while globulin rises to age 10 as a result of increased hepatic synthetic capacity and maturing immunity. Serum alkaline phosphatase activity peaks during bone growth spurts in infancy and adolescence due to osteoblast leakage, while creatinine increases with muscle mass. Serum gamma-glutamyl transferase, aspartate aminotransferase and lactate dehydrogenase activities are high at birth and decline with age. Serum alanine aminotransferase activity is low at birth and is induced by increased gluconeogenesis. Serum bilirubin increases continuously with age, mirroring haemoglobin concentration. Serum total cholesterol declines more markedly in boys than girls during puberty due to the combined effects of free testosterone (lowering high-density lipoprotein cholesterol in boys) and oestradiol (lowering low-density lipoprotein cholesterol in boys and girls).It is important to understand trends and biological variation when interpreting results since partitioned reference intervals may mask this information. PMID:26126034

  16. Biochemical analysis of serum and synovial fluid in clinically normal young camels (Camelus dromedarius

    Directory of Open Access Journals (Sweden)

    Raida Al-Rukibat

    2014-05-01

    Full Text Available The objective of this study was to determine the reference range values of various biochemical components in serum and synovial fluid in clinically normal young camels (Camelus dromedarius. One-hundred serum samples and 100 synovial fluid samples were collected from clinically, radiographically and cytologically normal carpal, tarsal and fetlock joints. The concentration of blood urea nitrogen (BUN, creatinine, glucose, sodium, calcium, magnesium, chloride, phosphorus, albumin and the activities of creatine kinase, alanine aminotransfearse, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatase (ALP were determined using commercially available kits. The concentration and activities of all measured parameters were significantly lower in the synovial fluid than in the serum except for the ALP and phosphorus, which were similar in both serum and synovial fluids. No significant difference was found in any of the measured biochemical parameters in different joints except in ALP activity, which was higher in the tarsal joint in comparison with the carpal and fetlock joint and the BUN concentration, which was higher in the tarsal joint in comparison with the carpal joint. Baseline values for biochemical components of normal camel synovial fluid and their serum counterparts have been generated. Such data can be used in the clinical investigation of camel’s joint diseases.

  17. 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 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)

  18. Characterization and site-directed mutagenesis of Wzb, an O-phosphatase from Lactobacillus rhamnosus

    Directory of Open Access Journals (Sweden)

    Gilbert Christophe

    2008-04-01

    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.

  19. Effect of Androctonus bicolor scorpion venom on the activities of serum enzymes in rats

    Science.gov (United States)

    Al-Asmari, Abdulrahman; Khan, Haseeb Ahmad; Manthiri, Rajamohammed Abbas

    2015-01-01

    We studied the effects of black fat-tailed scorpion (Androctonus bicolor) venom on the activities of liver enzymes including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), lactate dehydrogenase (LDH) and creatine kinase (CK) in the sera of rats. The animals were subcutaneously injected with a single dose of crude Androctonus bicolor venom (200 ?g/kg bodyweight) and were sacrificed at different time intervals including 30 min, 1 h, 2 h, 4 h, 8 h and 24 h after venom injection. There was no significant change in ALT activity in rats injected with Androctonus bicolor venom. Although Androctonus bicolor venom did not produce any change in serum AST activity until 1 h post-dosing, it significantly decreased this enzyme activity at 2 h onwards. There were significant decreases in ALP activities throughout the study though mild surges in the enzyme activity were observed at 1 h and 8 h post-dosing. There was a continued significant decrease in serum LDH activity until 8 h after Androctonus bicolor venom injection followed by normalization of LDH activity at 24 h. The activities of serum CK and GGT were significantly decreased at all the time points following Androctonus bicolor envenomation in rats. In conclusion, Androctonus bicolor envenomation in rats significantly reduced the activities of serum enzymes including AST, ALP, LDH, CK and GGT. Androctonus bicolor venom induced hypomagnesemia may account for persistently reduced activities of liver enzymes due to the cofactor role of magnesium in enzyme activities. PMID:26380012

  20. Application of Measurements of Serum CA15-3 and B-AKP in Diagnosis of Bone Metastasis in Patients with Post-operative Mammary Cancer

    International Nuclear Information System (INIS)

    To evaluate the diagnosis value of serum CA15-3 and B-AKP measurements in diagnosis of bone metastasis images in patients with post-operative mammary cancer, retrospective study on the bone scan images and serum CA15-3 and bone alkaline phosphatase (B-AKP) levels were performed in 92 patients with confirmed post-operative mammary gland cancer. The results showed that the serum levels of CA15-3 and B-AKP were increased step by step significantly along with the advancement of bone metastatic grading from M0 to M3 (P<0.01). The serum levels of CA15-3 and B-AKP were positively correlated with the number of bone metastasis. The positive rate of bone metastasis was 63.2% with serum CA15-3 more than 25U/mL; and the negative predictive value of bone metastasis was 94.5% with serum CA15-3 less than 25U/mL. The positive rate of bone metastasis was 59.6% with serum B-AKP levels more than 20U/L; and the negative predictive value of bone metastasis was 73.5% with serum B-AKP levels less than 20U/L. The negative predictive value of bone metastasis was 100% with serum CA15-3 less than 25U/mL and serum B-AKP levels less than 20U/L. The combined measurement of the serum CA15-3 and B-AKP levels would play an important role in diagnosis of bone scan images in patients with post-operative mammary cancer. (authors)

  1. Structural Genomics of Protein Phosphatases

    Energy Technology Data Exchange (ETDEWEB)

    Almo,S.; Bonanno, J.; Sauder, J.; Emtage, S.; Dilorenzo, T.; Malashkevich, V.; Wasserman, S.; Swaminathan, S.; Eswaramoorthy, S.; et al

    2007-01-01

    The New York SGX Research Center for Structural Genomics (NYSGXRC) of the NIGMS Protein Structure Initiative (PSI) has applied its high-throughput X-ray crystallographic structure determination platform to systematic studies of all human protein phosphatases and protein phosphatases from biomedically-relevant pathogens. To date, the NYSGXRC has determined structures of 21 distinct protein phosphatases: 14 from human, 2 from mouse, 2 from the pathogen Toxoplasma gondii, 1 from Trypanosoma brucei, the parasite responsible for African sleeping sickness, and 2 from the principal mosquito vector of malaria in Africa, Anopheles gambiae. These structures provide insights into both normal and pathophysiologic processes, including transcriptional regulation, regulation of major signaling pathways, neural development, and type 1 diabetes. In conjunction with the contributions of other international structural genomics consortia, these efforts promise to provide an unprecedented database and materials repository for structure-guided experimental and computational discovery of inhibitors for all classes of protein phosphatases.

  2. Serum fluoride and sialic acid levels in osteosarcoma.

    Science.gov (United States)

    Sandhu, R; Lal, H; Kundu, Z S; Kharb, S

    2011-12-01

    Osteosarcoma is a rare malignant bone tumor most commonly occurring in children and young adults presenting with painful swelling. Various etiological factors for osteosarcoma are ionizing radiation, family history of bone disorders and cancer, chemicals (fluoride, beryllium, and vinyl chloride), and viruses. Status of fluoride levels in serum of osteosarcoma is still not clear. Recent reports have indicated that there is a link between fluoride exposure and osteosarcoma. Glycoproteins and glycosaminoglycans are an integral part of bone and prolonged exposure to fluoride for long duration has been shown to cause degradation of collagen and ground substance in bones. The present study was planned to analyze serum fluoride, sialic acid, calcium, phosphorus, and alkaline phosphatase levels in 25 patients of osteosarcoma and age- and sex-matched subjects with bone-forming tumours other than osteosarcoma and musculo-skeletal pain (controls, 25 each). Fluoride levels were analyzed by ISE and sialic acid was analyzed by Warren's method. Mean serum fluoride concentration was found to be significantly higher in patients with osteosarcoma as compared to the other two groups. The mean value of flouride in patients with other bone-forming tumors was approximately 50% of the group of osteosarcoma; however, it was significantly higher when compared with patients of group I. Serum sialic acid concentration was found to be significantly raised in patients with osteosarcoma as well as in the group with other bone-forming tumors as compared to the group of controls. There was, however, no significant difference in the group of patients of osteosarcoma when compared with group of patients with other bone-forming tumors. These results showing higher level of fluoride with osteosarcoma compared to others suggesting a role of fluoride in the disease. PMID:19390788

  3. Modulation of antioxidant and phosphatase enzymes by beta-carotene against gamma radiation induced testicular disorders in albino rats

    International Nuclear Information System (INIS)

    Beta-carotene is a group of plant compounds called carotenoids. It is a precursor for vitamin A and an important antioxidant. This study aimed to evaluate the radioprotective efficacy of ?-carotene against gamma radiation induced disorders in the testis of male albino rats, it included 4 groups: control group, treated group; animals of this group received a daily oral dose of ?-carotene (30 mg/kg body wt) for 1 week, irradiated group; animals of this group were subjected to whole body gamma irradiation at a dose level of 6 Gy, and treated-irradiated group; animals received a daily oral dose of ?-carotene (30 mg/ kg body wt) for 1 week before exposure to whole body gamma irradiation at a dose level of 6 Gy. 6 animals of each group were autopsied at 1, 3 and 5 days after ?-carotene treatment and/ or irradiation. All animals were subjected to the following investigations: acid phosphatase, alkaline phosphatase, glutathione and glutathione peroxidase in testis homogenate. In irradiated animals there was a highly significant decrease in testis alkaline phosphatase, glutathione and glutathione peroxidase activity. On the other hand, significant increase in acid phosphatase activity was observed. Treatment with ?-carotene before irradiation causes significant increase in alkaline phosphatase, glutathione and glutathione peroxidase activity and significant decrease in acid phosphatase activity compared to the irradiated group. The results of the present study indicated that ?-carotene ameliorated oxidative stress and the loss of cellular antioxidants and suggest that ?-carotene may reduce the radiation damage in testis of male albino rats

  4. 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)

    JORGE ARTURO, REY CUBILLOS; LEONARDO, LAREO; SANDRA, GUTIÉRREZ; MARCELA, GODOY CORREDOR.

    2012-12-01

    Full Text Available 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 reaction (qRT-PCR). Aplication of AC increased expression of

  5. Glucose-6-phosphatase deficiency

    Directory of Open Access Journals (Sweden)

    Labrune Philippe

    2011-05-01

    Full Text Available Abstract Glucose-6-phosphatase deficiency (G6P deficiency, or glycogen storage disease type I (GSDI, is a group of inherited metabolic diseases, including types Ia and Ib, characterized by poor tolerance to fasting, growth retardation and hepatomegaly resulting from accumulation of glycogen and fat in the liver. Prevalence is unknown and annual incidence is around 1/100,000 births. GSDIa is the more frequent type, representing about 80% of GSDI patients. The disease commonly manifests, between the ages of 3 to 4 months by symptoms of hypoglycemia (tremors, seizures, cyanosis, apnea. Patients have poor tolerance to fasting, marked hepatomegaly, growth retardation (small stature and delayed puberty, generally improved by an appropriate diet, osteopenia and sometimes osteoporosis, full-cheeked round face, enlarged kydneys and platelet dysfunctions leading to frequent epistaxis. In addition, in GSDIb, neutropenia and neutrophil dysfunction are responsible for tendency towards infections, relapsing aphtous gingivostomatitis, and inflammatory bowel disease. Late complications are hepatic (adenomas with rare but possible transformation into hepatocarcinoma and renal (glomerular hyperfiltration leading to proteinuria and sometimes to renal insufficiency. GSDI is caused by a dysfunction in the G6P system, a key step in the regulation of glycemia. The deficit concerns the catalytic subunit G6P-alpha (type Ia which is restricted to expression in the liver, kidney and intestine, or the ubiquitously expressed G6P transporter (type Ib. Mutations in the genes G6PC (17q21 and SLC37A4 (11q23 respectively cause GSDIa and Ib. Many mutations have been identified in both genes,. Transmission is autosomal recessive. Diagnosis is based on clinical presentation, on abnormal basal values and absence of hyperglycemic response to glucagon. It can be confirmed by demonstrating a deficient activity of a G6P system component in a liver biopsy. To date, the diagnosis is most commonly confirmed by G6PC (GSDIa or SLC37A4 (GSDIb gene analysis, and the indications of liver biopsy to measure G6P activity are getting rarer and rarer. Differential diagnoses include the other GSDs, in particular type III (see this term. However, in GSDIII, glycemia and lactacidemia are high after a meal and low after a fast period (often with a later occurrence than that of type I. Primary liver tumors and Pepper syndrome (hepatic metastases of neuroblastoma may be evoked but are easily ruled out through clinical and ultrasound data. Antenatal diagnosis is possible through molecular analysis of amniocytes or chorionic villous cells. Pre-implantatory genetic diagnosis may also be discussed. Genetic counseling should be offered to patients and their families. The dietary treatment aims at avoiding hypoglycemia (frequent meals, nocturnal enteral feeding through a nasogastric tube, and later oral addition of uncooked starch and acidosis (restricted fructose and galactose intake. Liver transplantation, performed on the basis of poor metabolic control and/or hepatocarcinoma, corrects hypoglycemia, but renal involvement may continue to progress and neutropenia is not always corrected in type Ib. Kidney transplantation can be performed in case of severe renal insufficiency. Combined liver-kidney grafts have been performed in a few cases. Prognosis is usually good: late hepatic and renal complications may occur, however, with adapted management, patients have almost normal life span. Disease name and synonyms Glucose-6-phosphatase deficiency or G6P deficiency or glycogen storage disease type I or GSDI or type I glycogenosis or Von Gierke disease or Hepatorenal glycogenosis.

  6. Extracellular phosphatases of the Rimov reservoir phytoplankton

    International Nuclear Information System (INIS)

    Seasonal increases in extracellular phosphatase activity followed both phosphorus depletion and phytoplankton development in the Rimov reservoir during three consecutive seasons. A novel method, which enables localisation of extracellular phosphatases at the single-cell level using ELF phosphate, showed remarkable differences in phosphatase expression among particular phytoplankton species and also within one population. Phosphatase activity was detected in most of algal classes, except for Euglenophyceae, majority of Cryptophyceae and Chrysophyceae. Three algal species expressed extracellular phosphatases always when present in the phytoplankton, in contrast to many species that never expressed phosphatases. (authors)

  7. Serum biochemical and histopathological changes in liver and kidney in lambs after zinc oxide nanoparticles administration

    Directory of Open Access Journals (Sweden)

    H. Ganjealidarani

    2013-05-01

    Full Text Available Background: Zinc is an essential co-factor for many enzymatic activities and its deficiency may produce clinical signs of parakeratosis, loss and failure of growth of wool and hair in sheep. Aim: The present study was designed to evaluate the effect of zinc oxide nano-particles (instead conventional zinc oxide on serum biochemical factors and histopathological changes in liver and kidney of lambs. Materials and Methods: One group of lamb (4 lambs each group received zinc oxide nano-particles suspension daily for 25 day. Whole blood sample and serum were collected at the start and the end of study. Activities of lactate dehydrogenase (LDH, alanine aminotransferas (ALT, aspartate aminotransferase (AST, alkaline phosphatase (ALP, blood urea nitrogen (BUN, and creatinine (CR were measured in serum of lambs. Sections of liver and kidney were stained with hematoxylin-eosin and examined by light microscopy. Results: Activity of ALP (p=0.011 was significantly decreased and creatinine level (p=0.002 was significantly increased by zinc oxide nano-particles. Cell swelling, eosinophilic necrosis of hepatocytes, and multifocal interstitial nephritis was observed during histopatological examination. Conclusion: Results of study suggest that zinc oxide nano-particles may be toxic for use in sheep in zinc deficiency status. [Vet World 2013; 6(8.000: 534-537

  8. [False positive serum des-gamma-carboxy prothrombin after resection of hepatocellular carcinoma].

    Science.gov (United States)

    Hiramatsu, Kumiko; Tanaka, Yasuhito; Takagi, Kazumi; Iida, Takayasu; Takasaka, Yoshimitsu; Mizokami, Masashi

    2007-04-01

    Measurements of serum concentrations of des-gamma-carboxy-prothrombin (PIVKA-II) are widely used for diagnosing hepatocellular carcinoma (HCC). Recently, when we evaluated the correlation of PIVKA-II between two commercially available PIVKA-II immunoassay kits (Lumipulse f vs. Picolumi) to introduce it in our hospital, false high values of PIVKA-II were observed in Lumipulse assay. Four(4%) of 100 serum samples showed false high values, and all of them were obtained from patients less than 2 month after curative resection of HCC. Examining additional 7 patients with HCC resection, serum samples from the 5 patients had the same trend. To elucidate the non-specific reaction by Lumipulse assay which utilized alkaline phosphatase (ALP) enzymatic reaction, inhibition assays by various absorbents such as inactive ALP and IgM antibodies were performed. Excess of inactive ALP reduced the high values of PIVKA-II. Note that anti-bleeding sheets (fibrinogen combined drug), which included bovine thrombin, were directly attached on liver of all patients with HCC resection in this study. As the sheets also contaminate ALP and probably produce IgM antibodies to ALP, the IgM may cross-react with anti-PIVKA-II antibodies directly. Taken together, it was suggested that produced antibodies against ALP derived from anti-bleeding sheets led false high values of PIVKA-II in the patients with HCC resection. PMID:17511263

  9. The application of serum PSA and BALP measurements in bone scanning for metastasis in patients with prostate cancer diagnostic

    International Nuclear Information System (INIS)

    Objective: To evaluate the diagnosis value of serum PSA and BALP measurements for scanning bone metastatic images in patients with prostatic cancer. Methods: Retrospective study on the bone scan images and serum PSA (with CLIA) and bone alkaline phosphatase (BALP, with ELISA) levels were performed in 96 patients with confirmed prostatic cancer. Results: (1) The serum levels of PSA and BALP were increased step by step along with the advancement of bone metastatic grading from M0 to M3 with significant difference between values in successive gradings (P 20ng/ml, the positive rate of bone metastasis was 65.4%, with PSA 20u/L, the positive rate of bone metastasis was 58.9%, with BALP < 20u/L, the negative predictive value of bone metastasis was 76.5%. However with both PSA < 20ng/ml and BALP < 20u/L, the negative predictive value of bone metastasis was 100%. Conclusion: The combined measurement of the serum PSA and BALP levels would play an important role for diagnosis of bone scan images in patients with prostate cancer. (authors)

  10. Phosphate solubilization potential and phosphatase activity of rhizospheric Trichoderma spp.

    Scientific Electronic Library Online (English)

    Anil, Kapri; Lakshmi, Tewari.

    2010-10-01

    Full Text Available Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. [...] The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 µg.ml-1), followed by the standard culture of T. harzianum (386.42 µg.ml-1) after 96 h of incubation at 30+1(0)C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1) followed by the standard culture (13.41 U.ml-1) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth.

  11. Phosphate solubilization potential and phosphatase activity of rhizospheric Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Anil Kapri

    2010-10-01

    Full Text Available Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP broth containing tricalcium phosphate (TCP as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 µg.ml-1, followed by the standard culture of T. harzianum (386.42 µg.ml-1 after 96 h of incubation at 30+1(0C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP. High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1 followed by the standard culture (13.41 U.ml-1 at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP. Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth.

  12. Serum liver enzyme and histopathologic changes in calves with chronic and chronic-delayed Senecio jacobaea toxicosis.

    Science.gov (United States)

    Craig, A M; Pearson, E G; Meyer, C; Schmitz, J A

    1991-12-01

    Progressive changes in serum enzyme activity and liver histologic features were monitored in calves fed tansy ragwort (Senecio jacobaea)-contaminated pellets. The experiments were designed to simulate natural intoxicant ingestion conditions in relationship to the dose and duration of exposure to the toxic plant to correlate early laboratory diagnostic changes with the natural progression of the disease, thereby facilitating early diagnosis and intervention by veterinary clinicians. Eight calves were fed tansy ragwort and 4 additional calves served as controls. In group 1, 4 calves were continuously fed dried tansy ragwort mixed in a pelleted feed at a 5% concentration by dry weight until terminal liver disease developed. Serum liver enzyme (alkaline phosphatase, glutamate dehydrogenase, and gamma-glutamyltransferase) activities were monitored at weekly intervals in these calves and in the 2 controls. In group 2, 4 calves were fed the same contaminated feed for only 60 days, with return to normal feed for the duration of the trial. Two additional calves served as controls. Their liver enzyme activities were monitored every other week in conjunction with percutaneous liver biopsies. All 8 calves fed tansy ragwort-contaminated pellets developed terminal hepatopathy in either a chronic pattern (n = 6) or a chronic-delayed pattern (n = 2), with the onset of a moribund state or sudden death at 11 to 17 weeks and 27 to 51 weeks, respectively. The calves were euthanatized when classic terminal signs of hepatic encephalopathy first became evident. The clinicopathologic patterns of chronic and chronic-delayed toxicoses were typical of over 5,000 cases of field tansy toxicosis diagnosed at the diagnostic laboratory. Serum glutamate dehydrogenase was the first enzyme to increase in most animals, with a short-term increase to peak values followed by a rapid return to normal. This enzyme change was followed by increases in alkaline phosphatase and gamma-glutamyltransferase. Serum enzyme changes preceded development of recognizable histologic lesions. Vacuolar changes in hepatocyte nuclei, biliary hyperplasia, and fibrosis sequentially developed in liver biopsy specimens from each animal, whereas megalocytosis was not a predominant feature until necropsy. On the basis of our findings, we suggest that the optimal tests for diagnosis of pyrrolizidine alkaloid intoxication should consist of liver biopsy and determination of concurrent serum liver-enzyme activities. PMID:1686378

  13. Extracellular phosphatases in a Mediterranean reservoir: seasonal, spatial and kinetic heterogeneity.

    Czech Academy of Sciences Publication Activity Database

    Nedoma, Ji?í; García, J.C.; Comerma, M.; Šimek, Karel; Armengol, J.

    2006-01-01

    Ro?. 51, ?. 7 (2006), s. 1264-1276. ISSN 0046-5070 R&D Projects: GA ?R(CZ) GA206/99/0028 Grant ostatní: SICST(ES) HID96-1374-CO2; ICST(ES) 1997SGR-122 Institutional research plan: CEZ:AV0Z60170517 Keywords : alkaline phosphatase * eutrophication * P limitation Subject RIV: DJ - Water Pollution ; Quality Impact factor: 2.502, year: 2006

  14. Effect of Phosphatases Activity in the Hepatopancreas and Muscle of the Fresh Water Female Field Crab, Spiralothelphusa hydrodroma (Herbst Treated with Cypermethrin

    Directory of Open Access Journals (Sweden)

    R. S. Sreenivasan

    2011-04-01

    Full Text Available The fresh water field crab, Spiralothelphusa hydrodroma is an important human food source in parts of South India and the crab is constantly exposed to pesticides, which are used extensively to control agricultural pests. Evaluation of the toxic effect of cypermethrin on the experimental crab for the LC?? value was carried out. Effect of cypermethrin on the biochemical changes in the hepatopancreas and muscle was observed. Quantitative study of biochemical changes of acid phosphatase and alkaline phosphatase were undertaken.

  15. Uranium in alkaline rocks

    Energy Technology Data Exchange (ETDEWEB)

    Murphy, M.; Wollenberg, H.; Strisower, B.; Bowman, H.; Flexser, S.; Carmichael, I.

    1978-04-01

    Geologic and geochemical criteria were developed for the occurrence of economic uranium deposits in alkaline igneous rocks. A literature search, a limited chemical analytical program, and visits to three prominent alkaline-rock localities (Ilimaussaq, Greenland; Pocos de Caldas, Brazil; and Powderhorn, Colorado) were made to establish criteria to determine if a site had some uranium resource potential. From the literature, four alkaline-intrusive occurrences of differing character were identified as type-localities for uranium mineralization, and the important aspects of these localities were described. These characteristics were used to categorize and evaluate U.S. occurrences. The literature search disclosed 69 U.S. sites, encompassing nepheline syenite, alkaline granite, and carbonatite. It was possible to compare two-thirds of these sites to the type localities. A ranking system identified ten of the sites as most likely to have uranium resource potential.

  16. Uranium in alkaline rocks

    International Nuclear Information System (INIS)

    Geologic and geochemical criteria were developed for the occurrence of economic uranium deposits in alkaline igneous rocks. A literature search, a limited chemical analytical program, and visits to three prominent alkaline-rock localities (Ilimaussaq, Greenland; Pocos de Caldas, Brazil; and Powderhorn, Colorado) were made to establish criteria to determine if a site had some uranium resource potential. From the literature, four alkaline-intrusive occurrences of differing character were identified as type-localities for uranium mineralization, and the important aspects of these localities were described. These characteristics were used to categorize and evaluate U.S. occurrences. The literature search disclosed 69 U.S. sites, encompassing nepheline syenite, alkaline granite, and carbonatite. It was possible to compare two-thirds of these sites to the type localities. A ranking system identified ten of the sites as most likely to have uranium resource potential

  17. Advanced alkaline water electrolysis

    OpenAIRE

    MARINI, STEFANIA; SALVI, PAOLO; NELLI, PAOLO; PESENTI, RACHELE; Villa, Marco; Berrettoni, Mario; ZANGARI, GIOVANNI; Kiros, Yohannes

    2012-01-01

    A short review on the fundamental and technological issues relevant to water electrolysis in alkaline and proton exchange membrane (PEM) devices is given. Due to price and limited availability of the platinum group metal (PGM) catalysts they currently employ, PEM electrolyzers have scant possibilities of being employed in large-scale hydrogen production. The importance and recent advancements in the development of catalysts without PGMs are poised to benefit more the field of alkaline electro...

  18. Protective effect of irradiated licorice roots (Glycyrrhiza Glabra) on ethanol-induced serum lipid changes and liver Injury In rats

    International Nuclear Information System (INIS)

    Water extract of licorice roots irradiated with gamma rays at dose level of 20 KGy has been evaluated for hepato protective and hypolipidaemic effects against chronic ethanol mediated toxicity in rats.Ethanol administration to rats (7.9 g/kg/day) for 45 days resulted in significant elevated levels of serum total bilirubin, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total cholesterol (TC), triglycerides (TG), low density lipoprotein-cholesterol (LDL-C), thiobarbituric acid reactive substances (TBARS). Meanwhile, significant reductions in high density lipoprotein-cholesterol (HDL-C), total protein, albumin and glucose were noticed in comparison to those of control group, whereas, serum Na+ , K+ and globulin were kept unchanged. Co administration of raw and irradiated licorice roots (3 g/l in drinking water) with the daily dose of ethanol caused significant improvement in all the above mentioned parameters except serum glucose level. On the other hand, results showed that licorice roots water extract induced significant decrease in K+ level with increased Na+ level. There was non-significant difference between non-irradiated and irradiated licorice. Thus, it could be concluded that irradiated licorice roots has not affected its physiological functions. Moreover, supplementation with licorice roots water extract can offer protection against free radical mediated oxidative stress in hepatotoxicity

  19. Hematology and serum biochemistry of captive gharial (Gavialis gangeticus in India

    Directory of Open Access Journals (Sweden)

    Shahnaz Amin

    2014-10-01

    Full Text Available Aim: To study the hematological and serum biochemical parameters of the critically endangered gharial (Gavialis gangeticus. Materials and Methods: Blood samples for hemato-biochemical analyses were collected from the ventral median coccygeal vein of six juvenile and six sub adult gharials of Dewari Gharial Rearing Centre of National Chambal Sanctuary, Madhya Pradesh, India. Hematological examination was performed manually. Differential leukocyte count was performed on the blood smears stained with Giemsa’s stain. The analysis of serum was conducted by eppendorf ECOM-F 6124 semi auto biochemical analyzer using standard ERBA biochemical reagent kits. Results: Peripheral blood cells of gharial showed erythrocytes with an oval outline and centrally located prominent round to oval nucleus. Erythrocyte count in sub adult gharials was significantly greater than juveniles. Whereas erythrocyte mean corpuscular volume and erythrocyte size in juveniles was significantly larger than sub adults. The average most abundant leukocyte type in gharial was lymphocytes (53%, followed by heterophils (27%, eosinophils (10%, monocytes (7% and basophils (3%. Aspartate aminotransferase, alkaline phosphatase, blood urea nitrogen, triglycerides and albumin concentrations in sub adult gharials were significantly higher than juveniles. No significant differences were determined in other hemato-biochemical parameters between juvenile and sub adult gharials under study. Conclusion: A preliminary database on hematology and blood biochemistry of gharial was established. The data will be useful in routine health evaluations, especially in relation to determining potential effects associated with factors such as pollution and infectious diseases.

  20. The study of chemiluminescence immunoassay for determination of human serum true insulin

    International Nuclear Information System (INIS)

    To develop a highly sensitive CLIA to detect human serum true insulin, two specific monoclonal antibodies having different and distinct epitopes on insulin molecule were used in this study: one was coated on microtiter plate as the solid phase antibody and the other was labeled with alkaline phosphatase. Adamantine derivate was used as the substrate. The results showed that the sensitivity was 0.06 ?IU/mL, and the linear calibrator was in the range of 1.0-150 ?IU/mL. The CV of intra-and interbatches were 5.0% and 7.8%, respectively, and the mean recovery rate was 94.4%. According to measurement results of 200 samples (100 men and 100 women) the determination range was 0.52%-11.23 ?IU/mL for males, 0.75-10.66 ?IU/mL for females, and the mean value was 3.86, 3.62 ?IU/mL. There was no obvious difference between men and women. Chemiluminescence immunoassay (CLIA) is a simple and convenient, and can reflect truly the level of serum insulin. CLIA of insulin has application value in diagnosis and pathological research of diabetes mellitus. (authors)

  1. 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)

    ZHOU, DA-AN; DENG, YUE-NING; LIU, LEI; LI, JIAN-JUN

    2015-01-01

    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 (P<0.05). Furthermore, the proliferation following treatment with 25% KRMBTI-serum was significantly higher than that for the other KRMBTI-serum concentrations (P<0.01). For a 25% concentration of the serum collected at l h, the proliferation in the high- and low-dose KRMBTI-serum groups was significantly higher than that of the medium-dose and control groups (P<0.01) and no statistical significance was observed between the high- and low-dose groups. In the osteogenic differentiation process of the high-dose group, the ALP activity at every time-point was significantly higher than that of the low-dose group and the peak value of the former was achieved at concentrations between 20 and 30%. KRMBTI-serum was shown to promote the expression of TGF-?1. Furthermore, hepcidin 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

  2. Microrganismos do solo produtores de fosfatases em diferentes sistemas agrícolas Soil microorganisms phosphatase producers in different agricultural systems

    Directory of Open Access Journals (Sweden)

    Ely Nahas

    2002-12-01

    Full Text Available Considerando-se que uma forma de prover fosfato disponível para as plantas é através da atividade mineralizadora microbiana do fósforo orgânico, avaliou-se a influência da planta (braquiária, guandu e sem planta, dos fertilizantes (superfosfato simples, fosfato de rocha e sem adubo e da calagem (com e sem calcário nas populações de microrganismos produtores de fosfatases ácida e alcalina. Do total de bactérias, 70,6% apresentaram atividade de fosfatase alcalina e 58,2% de fosfatase ácida e dos fungos, 64,3% e 84,7% respectivamente. Esses dados mostram número significativo de microrganismos com habilidade de mineralização do fósforo orgânico. Observou-se efeito restritivo do guandu sobre as bactérias e fungos produtores de fosfatases alcalina e ácida, cujos números foram sempre inferiores aos obtidos com a cultura de braquiária ou com as parcelas sem cultivo. Maior número de bactérias produtoras de fosfatase alcalina foi obtido nos tratamentos com superfosfato e controle do que com fosfato de rocha. Ao contrário, para os fungos, encontrou-se maior número nas parcelas não fertilizadas que as adubadas com fosfato. O número de fungos com atividade de fosfatase ácida diminuiu por efeito da calagem, enquanto o das bactérias aumentou. Finalmente, o número de bactérias produtoras de fosfatase alcalina superou o de fungos.Considering that one way for providing available phosphate to plants is by the microbial activity for the mineralization of organic phosphorus, the influence of plants (Brachiaria ruziziensis, Cajanus cajan and control, fertilizers (superphosphate, rock phosphate and control and liming (with and without lime was evaluated in the microorganisms populations of the acid and alkaline phosphatase producers. Among bacteria, 70,6% showed alkaline phosphatase activity and 58,2% acid phosphatase activity and among fungi, 64,3% and 84,7%, respectively. These numbers show that a significant population of microorganisms shows ability for organic phosphorus mineralization. Restrictive effect of the C. cajan was observed on the bacteria and fungi producers of the alkaline and acid phosphatases whose numbers always were lower to those obtained with the B. ruziziensis or with the plots without cultivation. Larger number of alkaline phosphatase bacterial producers was obtained in the treatments with superphosphate and control than that with rock phosphate. On the opposite, for fungi, a larger number was found in the plots with no fertilizer compared to the ones with fertilizer. The population of fungi having acid phosphatase activity decreased in the limed plots whereas the bacterial population increased. Finally, the alkaline phosphatase bacterial producers showed higher enzymatic activity than those of the fungi.

  3. Microrganismos do solo produtores de fosfatases em diferentes sistemas agrícolas / Soil microorganisms phosphatase producers in different agricultural systems

    Scientific Electronic Library Online (English)

    Ely, Nahas.

    2002-12-01

    Full Text Available Considerando-se que uma forma de prover fosfato disponível para as plantas é através da atividade mineralizadora microbiana do fósforo orgânico, avaliou-se a influência da planta (braquiária, guandu e sem planta), dos fertilizantes (superfosfato simples, fosfato de rocha e sem adubo) e da calagem (c [...] om e sem calcário) nas populações de microrganismos produtores de fosfatases ácida e alcalina. Do total de bactérias, 70,6% apresentaram atividade de fosfatase alcalina e 58,2% de fosfatase ácida e dos fungos, 64,3% e 84,7% respectivamente. Esses dados mostram número significativo de microrganismos com habilidade de mineralização do fósforo orgânico. Observou-se efeito restritivo do guandu sobre as bactérias e fungos produtores de fosfatases alcalina e ácida, cujos números foram sempre inferiores aos obtidos com a cultura de braquiária ou com as parcelas sem cultivo. Maior número de bactérias produtoras de fosfatase alcalina foi obtido nos tratamentos com superfosfato e controle do que com fosfato de rocha. Ao contrário, para os fungos, encontrou-se maior número nas parcelas não fertilizadas que as adubadas com fosfato. O número de fungos com atividade de fosfatase ácida diminuiu por efeito da calagem, enquanto o das bactérias aumentou. Finalmente, o número de bactérias produtoras de fosfatase alcalina superou o de fungos. Abstract in english Considering that one way for providing available phosphate to plants is by the microbial activity for the mineralization of organic phosphorus, the influence of plants (Brachiaria ruziziensis, Cajanus cajan and control), fertilizers (superphosphate, rock phosphate and control) and liming (with and w [...] ithout lime) was evaluated in the microorganisms populations of the acid and alkaline phosphatase producers. Among bacteria, 70,6% showed alkaline phosphatase activity and 58,2% acid phosphatase activity and among fungi, 64,3% and 84,7%, respectively. These numbers show that a significant population of microorganisms shows ability for organic phosphorus mineralization. Restrictive effect of the C. cajan was observed on the bacteria and fungi producers of the alkaline and acid phosphatases whose numbers always were lower to those obtained with the B. ruziziensis or with the plots without cultivation. Larger number of alkaline phosphatase bacterial producers was obtained in the treatments with superphosphate and control than that with rock phosphate. On the opposite, for fungi, a larger number was found in the plots with no fertilizer compared to the ones with fertilizer. The population of fungi having acid phosphatase activity decreased in the limed plots whereas the bacterial population increased. Finally, the alkaline phosphatase bacterial producers showed higher enzymatic activity than those of the fungi.

  4. Protein kinase and phosphatase activities of thylakoid membranes

    International Nuclear Information System (INIS)

    Dephosphorylation of the 25 and 27 kDa light-harvesting Chl a/b proteins (LHCII) of the thylakoid membranes is catalyzed by a phosphatase which differs from previously reported thylakoid-bound phosphatases in having an alkaline pH optimum (9.0) and a requirement for Mg2+ ions. Dephosphorylation of the 8.3 kDa psb H gene product requires a Mg2+ ion concentration more than 200 fold higher than that for dephosphorylation of LHC II. The 8.3 kDa and 27 kDa proteins appear to be phosphorylated by two distinct kinases, which differ in substrate specificity and sensitivity to inhibitors. The plastoquinone antagonist 2,5-dibromo-3-methyl-6-isopropyl-benzoquinone (DBMIB) inhibits phosphorylation of the 27 kDa LHC II much more readily than phosphorylation of the 8.3 kDa protein. A similar pattern of inhibition is seen for two synthetic oligopeptides (MRKSATTKKAVC and ATQTLESSSRC) which are analogs of the phosphorylation sites of the two proteins. Possible modes of action of DBMIB are discussed. 45 refs., 7 figs., 3 tabs

  5. Diagnostic value of prostatic acid phosphatase as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Serum concentrations of prostatic acid phosphatase (PAP) were determined with 4 different radioimmunoassays and with the standard enzymatic method (p-nitrophenylphosphate) in 35 patients with prostatic carcinoma. Staging of localized tumors was based on histopathological evaluation after radial prostatectomy and pelvic lymphnode dissection (pTsub(1-3), pN0). In tumor lesions Tsub(1-2) N0 M0 elevated PAP-serum concentrations were found by RIA-determination in only one patient. Increased PAP serum levels were observed in 43-78% of carcinomas stage T3 N0 M0 and in 54-83% in stage Tsub(2-4) Nsub(x) M1 tumors, depending on the test kit used for the PAP determination. Concentrations for PAP obtained with the 4 different RIA-kits used, varied significantly and thus are not comparable. No false positive results were observed in sera of 9 patients with benign prostatic hyperplasia. Elevated PAP serum levels were found in a significantly higher frequency when determined by radioimmunoassay than by the enzymatic method. The results clearly indicate, that PAP is of no value for early recognition of carcinoma of the prostate even when measured by radioimmunoassay. However, the RIA-method seems to be of clinical importance in estimating the course of advanced local and metastasizing carcinoma of the prostate. (orig.)

  6. Advanced alkaline water electrolysis

    International Nuclear Information System (INIS)

    A short review on the fundamental and technological issues relevant to water electrolysis in alkaline and proton exchange membrane (PEM) devices is given. Due to price and limited availability of the platinum group metal (PGM) catalysts they currently employ, PEM electrolyzers have scant possibilities of being employed in large-scale hydrogen production. The importance and recent advancements in the development of catalysts without PGMs are poised to benefit more the field of alkaline electrolysis rather than that of PEM devices. This paper presents our original data which demonstrate that an advanced alkaline electrolyzer with performances rivaling those of PEM electrolyzers can be made without PGM and with catalysts of high stability and durability. Studies on the advantages/limitations of electrolyzers with different architectures do show how a judicious application of pressure differentials in a recirculating electrolyte scheme helps reduce mass transport limitations, increasing efficiency and power density.

  7. The application of radionuclide bone scintigraphy, serum PSA and ALP measurement in diagnosis of bone metastasis in prostate cancer patients

    International Nuclear Information System (INIS)

    Objective: To evaluate the clinical value of radionuclide bone scintigraphy, serum prostate-specific antigen (PSA) and alkaline phosphatase (ALP) measurement in the diagnosis of bone metastasis(BM) in prostate cancer patients. Methods: The results of bone scans, serum PSA and ALP levels were reviewed in 37 patients with prostate cancer. Correlation analysis was performed between PSA, ALP levels and BM grade. Results: The incidence rate of osseous metastasis was 70.3% (26/37), the most common involved parts were spine and pelvis. The serum PSA level in 18 untreated patients with BM was all >20 ng/ml, which was significantly different with those in 9 untreated patients without BM(NBM) (P0.05), but significant difference could be received when the cutoff value is 0.4 ng/ml(P>0.05). There was a positive correlation between serum ALP levels and BM degree(r=0.752, P=0.01). The levels of PSA in untreated and treated patients also had positive correlation with BM grade (r=0.508, P=0.01; r=0.515, P=0.05). Conclusion: Radionuclide bone scintigraphy is major method in diagnosis of bone metastasis in prostate cancer patients currently. The levels of PSA ?20% ng/ml in Patients with newly diagnosed and untreated prostate cancer should undergo bone scintigraphy. For those treated patients, bone scintigraphy should be performed when the PSA is ?0.4 ng/ml. The diagnostic efficacy of ALP is better than that of PSA in untreated patients. The level of serum ALP is also correlates with the degree of BM. (authors)

  8. The Influence of Sunlight Exposure on Serum Vitamin D Concentration and Bone Turnover; a controlled clinical trial

    Directory of Open Access Journals (Sweden)

    A Ataie-Jafari

    2008-11-01

    Full Text Available "nBackground: Sunlight exposure is one of the ways for vitamin D synthesis. However, its effect on vitamin D status via experimental studies is poorly understood. This study was undertaken to address the possibility that sunlight exposure may increase the levels of serum vitamin D, and alter bone turnover in healthy young girls."nMethods: In a controlled clinical trial, young girls were assigned to the test group (n= 45 or control group (n= 80. An out­door swimming pool was considered for this project and the test group was required to participate in these sessions at least for 8 sessions and to expose to direct sunlight at least for 20 minutes in each session. They were not allowed to use sun­screen during this time. Control group continued their usual manner of sun exposing. Serum levels of vitamin D, calcium, alkaline phosphatase, parathormone, osteocalcin and crossLaps were measured before and after duration of the study in both groups and compared between them."nResults: Subjects aged 27.46±8.78 years. Serum levels of vitamin D and bone markers were constant during the study in both groups. Changes of these variables were not significant between the groups after the study. Serum vitamin D in sub­jects with white skin color correlated with total time of direct sun exposing after the study (P= 0.002."nConclusion: Sunlight exposure did not affect the serum vitamin D and bone turnover in healthy young girls. However, sub­jects with bright skin complexion benefit from sunlight exposing more than those with a dark skin color in the case of vita­min D improvement.  

  9. Level of serum 25-OHD in healthy children aged 0-36 months in Van - Original Article

    Directory of Open Access Journals (Sweden)

    Vefik Ar?ca

    2010-09-01

    Full Text Available Aim: In recent studies, it has been shown that prevalence of rachitism and vitamin D deficiency depend on regional differences such as climate, socioeconomic level and changing benefits of people from health services. Even if no clinical symptom has occurred, serum 25-hydroxy D (25-OHD level, which is the best indicator of vitamin D can be found low. Material and Method: In this study, serum 25-OHD levels of 112 healthy children, aging 0-36 months, who applied to the outpatient clinic of the Pediatrics and Gynecology Hospital for a routine control in Van, were analyzed. Nutrition style of mothers and their babies, duration of exposing to sunlight and taken vitamin supplements, were evaluated. Serum Ca, P, alkaline phosphatase and 25-OHD levels were studied and the left wrist x-rays were obtained. Abdominal ultrasonography was performed only the babies with serum 25-OHD level >150 ng/mL. Results: In our study, despite no clinical symptoms of rachitism regardless of gender, 25-OHD level <40 ng/mL was determined in 53.5% of the children and in %13,3 of these childrens serum level of 25-OHD was as low as <5 ng/mL, and suffering from heavy vitamin D deficieny. Conclusions: The breast-fed babies with no vitamin supplement did not show any sign of vitamin deficiency, but in 25-OHD levels were significantly low compared to the breast-fed babies with vitamin supplement. (Turk Arch Ped 2010; 45: 286-90

  10. P depletion and activity of phosphatases in the rhizosphere of mycorrhizal and non-mycorrhizal cucumber (Cucumis Sativus L.)

    DEFF Research Database (Denmark)

    Joner, E.J.; Magid, J.

    1995-01-01

    An experiment was set up to test the ability of arbuscular mycorrhizal (AM) roots and hyphae to produce extracellular phosphatases and to study the relationship between phosphatase activity and soil organic P (P-o). Non-mycorrhizal cucumber and cucumber in symbiosis with either of two mycorrhizal fungi were grown in a sandy loam-sand mixture in three-compartment pots. Plant roots were separated from two consecutively adjoining compartments, first by a 37 m mesh excluding roots and subsequently by a 0.45 m membrane excluding mycorrhizal hyphae. Soil from the two root-free compartments was sectioned in a freezing microtome and analyzed for extracellular acid (pH 5.2) and alkaline (pH 8.5) phosphatase activity as well as depletion of NaHCO-3-extractable inorganic P (P-i) and P-o. Roots and mycorrhizal hyphae depleted the soil of P-i but did not influence the concentration of P-o in spite of increased phosphatase activity in soil influenced by roots. Phosphatase activity at both pH values was highest in soil influenced by uncolonized roots, but this was attributed to higher root length densities as compared to mycorrhizal roots. Mycorrhizal hyphae showed no influence on soil phosphatase activity in spite of high hyphal length densities ( gt 22 m cm-3). Hyphae were also able to deplete soil of P-i beyond the membrane interface.

  11. Biomineralization of Uranium by PhoY Phosphatase Activity Aids Cell Survival in Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Yung, M C [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Jiao, Y [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2014-07-22

    Caulobacter crescentus is known to tolerate high levels of uranium [U(VI)], but its detoxification mechanism is poorly understood. Here we show that C. crescentus is able to facilitate U(VI) biomineralization through the formation of U-Pi precipitates via its native alkaline phosphatase activity. The U-Pi precipitates, deposited on the cell surface in the form of meta-autunite structures, have a lower U/Pi ratio than do chemically produced precipitates. The enzyme that is responsible for the phosphatase activity and thus the biomineralization process is identified as PhoY, a periplasmic alkaline phosphatase with broad substrate specificity. Furthermore, PhoY is shown to confer a survival advantage on C. crescentus toward U(VI) under both growth and nongrowth conditions. Results obtained in this study thus highlight U(VI) biomineralization as a resistance mechanism in microbes, which not only improves our understanding of bacterium-mineral interactions but also aids in defining potential ecological niches for metal-resistant bacteria.

  12. Detection of extracellular phosphatase activity at the single-cell level by Enzyme-Labeled Fluorescence and flow cytometry: The importance of time kinetics in ELFA labeling.

    Czech Academy of Sciences Publication Activity Database

    Duhamel, S.; Gregori, G.; Van Wambeke, F.; Nedoma, Ji?í

    75A, ?. 2 (2009), s. 163-168. ISSN 1552-4922 Grant ostatní: MŠMT(CZ) PAI Barrande 2005-06-009-01 Institutional research plan: CEZ:AV0Z60170517 Keywords : alkaline phosphatase * ELF phosphate * heterotrophic bacteria Subject RIV: DA - Hydrology ; Limnology Impact factor: 3.032, year: 2009

  13. Fasted and postprandial response of serum physiological response, hepatic antioxidant abilities and HSP70 expression in Wuchang bream (Megalobrama amblycephala fed different dietary carbohydrate levels

    Directory of Open Access Journals (Sweden)

    Chuanpeng Zhou

    2014-12-01

    Full Text Available The effect of dietary carbohydrate (CHO level on serum physiological response, hepatic antioxidant abilities and heat shock protein 70 (HSP70 expression of Wuchang bream (Megalobrama amblycephala was studied. Two isonitrogenous (28.56% crude protein and isolipidic (5.28% crude lipid diets were formulated to contain 30% or 53% wheat starch. Diets were fed for 90 days to fish in triplicate tanks (28 fish per tank. At the end of feeding trial, significantly higher serum triglyceride level, insulin level, cortisol level, and malondialdehyde (MDA content were observed in fish fed the 53% CHO diet, while significantly lower serum total protein content, alkaline phosphatase activity, superoxide dismutase activity and total antioxidative capacity were found in fish fed the 53% CHO diet compared with those fed the 30% diet. The relative level of hepatic heat shock protein 70 mRNA was significantly higher in the 53% CHO group than that in the 30% CHO at 6, 12 and 48 h after feeding. Ingestion of 53% dietary CHO impacts the nonspecific immune ability and causes metabolic stress in Megalobrama amblycephala.

  14. Fasted and postprandial response of serum physiological response, hepatic antioxidant abilities and HSP70 expression in Wuchang bream (Megalobrama amblycephala) fed different dietary carbohydrate levels

    Scientific Electronic Library Online (English)

    Chuanpeng, Zhou; Xianping, Ge; Bo, Liu; Jun, Xie; Ruli, Chen.

    2014-12-01

    Full Text Available The effect of dietary carbohydrate (CHO) level on serum physiological response, hepatic antioxidant abilities and heat shock protein 70 (HSP70) expression of Wuchang bream (Megalobrama amblycephala) was studied. Two isonitrogenous (28.56% crude protein) and isolipidic (5.28% crude lipid) diets were [...] formulated to contain 30% or 53% wheat starch. Diets were fed for 90 days to fish in triplicate tanks (28 fish per tank). At the end of feeding trial, significantly higher serum triglyceride level, insulin level, cortisol level, and malondialdehyde (MDA) content were observed in fish fed the 53% CHO diet, while significantly lower serum total protein content, alkaline phosphatase activity, superoxide dismutase activity and total antioxidative capacity were found in fish fed the 53% CHO diet compared with those fed the 30% diet. The relative level of hepatic heat shock protein 70 mRNA was significantly higher in the 53% CHO group than that in the 30% CHO at 6, 12 and 48 h after feeding. Ingestion of 53% dietary CHO impacts the nonspecific immune ability and causes metabolic stress in Megalobrama amblycephala.

  15. Nutrition and immunity: the effects of the combination of arginine and tryptophan on growth performance, serum parameters and immune response in broiler chickens challenged with infectious bursal disease vaccine.

    Science.gov (United States)

    Emadi, M; Jahanshiri, F; Kaveh, K; Hair-Bejo, M; Ideris, A; Alimon, A R

    2011-02-01

    To explore the effects of the combination of tryptophan (Trp) and arginine (Arg) on growth performance, serum parameters and immune response of broiler chickens challenged with intermediate plus strain of infectious bursal disease virus vaccine, an in vivo experiment was conducted. A corn-soybean meal-based diet containing different levels of Arg and Trp was used. Cobb500 male broiler chickens from 0 to 49 days of age were subjected to a diet supplemented with the combination of Trp and Arg. Growth performance parameters and serum parameters were measured at 27 and 49 days of age. To evaluate the immunomodulatory effects of the combination of Trp and Arg on the challenged chickens, we measured the serum levels of interferon-?, interferon-? and immunoglobulin G at 27, 35, 42, and 49 days of age. The results showed that the three evaluated immune system parameters including interferon-?, interferon-? and immunoglobulin G were significantly enhanced after treatment. This enhancement resulted in the recovery of infectious bursal disease virus-infected chickens compared with controls as confirmed by histopathological examinations. Moreover, serum parameters such as albumin and total protein increased, whereas the treatment decreased (P<0.05) the feed:gain ratio, aspartate amino-transferase, alkaline phosphatase, lactic dehydrogenase, triglyceride and cholesterol. These findings suggest that the combination of Arg and Trp has a regulatory effect on growth performance. Moreover, it modulates the systemic immune response against infectious bursal disease. PMID:21331949

  16. Structure, mapping, and expression of erp, a growth factor-inducible gene encoding a nontransmembrane protein tyrosine phosphatase, and effect of ERP on cell growth.

    OpenAIRE

    Noguchi, T.; Metz, R.; Chen, L; Mattéi, M G; Carrasco, D.; Bravo, R.

    1993-01-01

    We have characterized a growth factor-inducible gene, erp, and demonstrated that it encodes a 367-amino-acid nontransmembrane tyrosine phosphatase protein with significant similarity to the vaccinia virus H1 protein. Immunoprecipitation analyses show that the erp protein, ERP, is rapidly induced following serum stimulation of quiescent fibroblasts. ERP has been expressed as a fusion protein with glutathione S-transferase and shown to have tyrosine as well as serine protein phosphatase activit...

  17. Influence of VAM Fungi, Azotobacter sp. and PSB on Soil Phosphatase Activity and Nutrients (N, P, K, Cu, Zn, Fe and Mn) Status in the Rhizosphere of Stevia rebaudiana (Bert.) Plants

    OpenAIRE

    G. Ramakrishnaiah; T. Vijaya

    2013-01-01

    The pot culture experiment was carried out to evaluate the influence of different combinations of microbial inoculants (VAM + AZO + PSB) on soil phosphatase activity andnutrients (N, P, K, Cu, Zn, Fe, Cu & Mn) of Steviarebaudia plant rhizosphere on 20th, 40th and 60th day. The study revealed that the combined application of microbial inoculants onsoil Alkaline and Acid phosphatase activity has been found maximum at 60th day in triple inoculated plants than dual inoculated and control. The re...

  18. Alkaline quinone flow battery.

    Science.gov (United States)

    Lin, Kaixiang; Chen, Qing; Gerhardt, Michael R; Tong, Liuchuan; Kim, Sang Bok; Eisenach, Louise; Valle, Alvaro W; Hardee, David; Gordon, Roy G; Aziz, Michael J; Marshak, Michael P

    2015-09-25

    Storage of photovoltaic and wind electricity in batteries could solve the mismatch problem between the intermittent supply of these renewable resources and variable demand. Flow batteries permit more economical long-duration discharge than solid-electrode batteries by using liquid electrolytes stored outside of the battery. We report an alkaline flow battery based on redox-active organic molecules that are composed entirely of Earth-abundant elements and are nontoxic, nonflammable, and safe for use in residential and commercial environments. The battery operates efficiently with high power density near room temperature. These results demonstrate the stability and performance of redox-active organic molecules in alkaline flow batteries, potentially enabling cost-effective stationary storage of renewable energy. PMID:26404834

  19. Alkaline reaction powder concretes

    Directory of Open Access Journals (Sweden)

    Aleksandr Alekseevich Shishkin

    2014-02-01

    Full Text Available Current state of construction science causes during the construction of unique buildings, and construction of complex structures and their repair, high-strength binders and concretes based on them. High-strength concrete appeared in foreign practice in the early 60-ies different countries on an industrial scale started using concrete strength greater than 40 MPa. Particularly promising obtained at the end of the 80-ies of the twentieth century, the so-called reactive powder concrete - Reactive powder concretes (RPC. Concrete powder as reaction components due to the high dispersity and increased amounts of hydraulically active materials. At the same time, there exists a long form as slag-alkaline cementations binder, the activity of which, even without the use of special techniques used to improve the strength portland cement concrete, described above, up to 80 MPa. Fixed effect of the interaction between sodium silicate and iron salts and the resulting so-called slag slurry binder is a mixture of granulated blast furnace slag waste mining and processing (iron-bearing mineral complex, mixed with water. These two positions were the basis for a new type of concrete, a so-called slag-alkaline reactive powder concrete, which is a mixture of granulated blast furnace slag to iron- mineral complex, mixing an aqueous solution of the alkaline component with the addition of a polyalcohol. This type of binder has a compressive strength reaching 110 MPa.

  20. Serum Proteins and Some Biochemical Parameters in Broiler Chickens Fed with Raw and Treated Bitter Vetch (Vicia ervilia Seeds

    Directory of Open Access Journals (Sweden)

    Gh. Sadeghi

    2007-01-01

    Full Text Available This study carried out to evaluate the effect of bitter vetch seeds on serum proteins and biochemical parameters in broiler chickens. A total of 1320 one-day old broiler chicks of a commercial breed were placed in 64 pens. Treatments were included raw and four different processed bitter vetch seeds in three levels (150, 300 and 450 g kg-1 and a corn-soybean based diet as control. Each treatment group consisted of four replicates. Processing methods were included soaked in water for 12 h, autoclaved, then dried at room temperature (SAD; ground, soaked in water for 24 h, autoclaved and dried (GSAD; ground, soaked in water for 47 h with exchange water every 12 h, cooked and dried (GSCD and ground, soaked at 1% acetic acid solution for 24 h at 60°C (AA. Feeding raw, AA and GSAD seeds decreased serum albumin significantly (p<0.05 in 21 days old chicks. Chickens that fed with raw and treated bitter vetch seed had lower ?1 and ? globulins than control (p<0.05. Increasing raw and treated bitter vetch seeds from 15 to 30 and 45% decreased albumin, ?1 and ? globulins and increased ?2 and ? globulins significantly (p<0.05. In 14 days old chicks feeding raw and treated biter vetch had no effect on serum urea, but uric acid concentration decreased significantly (p<0.05. Feeding SAD seeds increased serum urea significantly (p<0.05, but uric acid concentration did not change with feeding raw and treated bitter vetch seeds in 42 day old chicks. Adding raw and treated bitter vetch seeds to diet increased T4 and decreased T3 concentrations in all ages. At 28 days old chicks, feeding raw and treated biter vetch seeds decreased alkaline phosphatase concentration significantly than control. Results showed that raw bitter vetch seeds have some toxic effects on metabolism in broiler chickens and GSCD and SAD treatments were more effective to detoxification of this seed.

  1. Dickkopf-1 and sclerostin serum levels in patients with systemic mastocytosis.

    Science.gov (United States)

    Rossini, Maurizio; Viapiana, Ombretta; Zanotti, Roberta; Tripi, Gaia; Perbellini, Omar; Idolazzi, Luca; Bonifacio, Massimiliano; Adami, Silvano; Gatti, Davide

    2015-05-01

    Bone involvement, mainly osteoporosis but also osteosclerosis, is frequent in patients with indolent systemic mastocytosis (ISM). The recent characterization of the canonical Wnt/?-catenin pathway in the regulation of bone remodeling provided important insights for our understanding of the pathophysiology of a number of conditions. The regulation of Wnt pathway in bone is predominantly driven by the production of receptor inhibitors such as Dickkopf-1 (DKK1) and sclerostin (SOST). This study aimed to explore if the various bone involvements in patients with ISM might be explained by variations in serum levels of DKK1 and SOST. This is a cross-sectional study in an adult ISM cohort (13 men and 13 women with diagnosed ISM) and fifty-two healthy sex and age-matched controls. Early morning, fasting and venous sampling was obtained in all subjects. The main outcome measures were serum bone-specific alkaline phosphatase (bALP), C-terminal telopeptides of type I collagene (CTX), DKK1, SOST, parathyroid hormone (PTH), bone mineral density, and prevalent vertebral fractures. Mean DKK1 serum levels were about two-folds higher in patients, than in controls (65,0 ± 43.3 vs. 33.1 ± 19.4 pmol/L, respectively; p < 0.001), irrespective of the presence of osteoporotic or diffuse osteosclerotic bone involvement. DKK1 serum levels were positively correlated with PTH and both CTX and bALP. Mean SOST serum levels were not significantly different in patients versus controls, and we did not observe any significant correlation between SOST and any available clinical or laboratory parameters, with the only exception of a positive correlation with age. In conclusion, in our study, we observed that DKK1, but not SOST, serum levels significantly increased in ISM patients with various bone involvements, and correlated with PTH and bone turnover markers. Our results suggest that the Wnt/?-catenin pathway is not primarily involved in the pathophysiology of the array of bone involvement in ISM. PMID:25694360

  2. Analysis of Smad Phosphatase Activity In Vitro.

    Science.gov (United States)

    Shen, Tao; Qin, Lan; Lin, Xia

    2016-01-01

    Phosphorylation of Smad1/5/8 at the C-terminal SXS motif by BMP type I receptors is one of the most critical events in BMP signaling. Conversely, protein phosphatases that dephosphorylate phospho-Smad1/5/8 can consequently prevent or terminate BMP signaling. PPM1H is an undercharacterized phosphatase in the PPM family. We recently demonstrated that PPM1H can dephosphorylate Smad1 in the cytoplasm and block BMP signaling responses in cellular assays. Here we describe in vitro method showing that PPM1H is a bona fide phosphatase for Smad1/5/8. PPM1H is produced as GST fusion protein in E. coli, and purified against glutathione sepharose beads. Bacterially purified recombinant PPM1H possesses phosphatase activity toward artificial substrate para-nitrophenyl phosphate (pNPP). Recombinant PPM1H also dephosphorylates immuno-purified phosphorylated Smad1 in test tubes. These direct in vitro phosphatase assays provide convincing evidence demonstrating the role of PPM1H as a specific phosphatase for P-Smad1. PMID:26520120

  3. Measuring Activity and Specificity of Protein Phosphatases.

    Science.gov (United States)

    Powers, Brendan L; Melesse, Michael; Eissler, Christie L; Charbonneau, Harry; Hall, Mark C

    2016-01-01

    Reversible protein phosphorylation plays essential roles in coordinating cell division and many other biological processes. Cell cycle regulation by opposing kinase and protein phosphatase activities is often complex and major challenges exist in identifying the direct substrates of these enzymes and the specific sites at which they act. While cell cycle kinases are known to exhibit strict substrate specificities important for coordinating the complex events of cell division, phosphatases have only recently been recognized to exert similarly precise regulatory control over cell cycle events through timely dephosphorylation of specific substrates. The molecular determinants for substrate recognition by many phosphatases that function in cell division are still poorly delineated. To understand phosphatase specificity, it is critical to employ methods that monitor the dephosphorylation of individual phosphorylation sites on physiologically relevant substrates. Here, using the cell cycle phosphatase Cdc14 as an example, we describe two methods for studying phosphatase specificity, one using synthetic phosphopeptide substrates and the other using intact phosphoprotein substrates. These methods are useful for targeted characterization of small substrate sets and are also adaptable to large-scale applications for global specificity studies. PMID:26254927

  4. Serum Calcium Levels in Newly-diagnosed Patients with Tuberculosis in Hamedan (West of Iran

    Directory of Open Access Journals (Sweden)

    Seyyed Hamid Hashemi

    2005-01-01

    Full Text Available To investigate the incidence of hypercalcemia in TB patients and to assess its relationship with TB, a study was conducted in Infectious Diseases Division of Sina Hospital in Hamedan province (west of Iran. During an 18-month period, 65 patients with newly-diagnosed TB (30 males and 35 females, were prospectively evaluated who aged between 15 and 84 years (mean 53.3 years. Age- and sex-matched subjects (82 with chronic obstructive pulmonary disease (27 males and 55 females, aged between 16 and 85 (mean 53.5 years was selected as a control group. Serum calcium, phosphorus, total protein, albumin and alkaline phosphatase were measured in all subjects. No significant difference was found between the mean albumin-adjusted calcium levels in the TB group (8.79±1.53 mg dL-1 and the control group (8.57±1.12 mg dL-1. Hypercalcemia was found in 11 (16.9 % of TB patients and 6 (7.3 % of controls (?2 = 3.27, p = 0.07, non significant. This study revealed that there is non significant difference association between hypercalcemia and tuberculosis.

  5. [Evaluation of serum PIVKA-II by Lumipulse PrestoII assay].

    Science.gov (United States)

    Hiramatsu, Kumiko; Tanaka, Yasuhito; Takagi, Kazumi; Kani, Satomi; Goto, Takaaki; Takasaka, Yoshimitsu; Matsuura, Kentaro; Sugauchi, Fuminaka; Moriyama, Kazushige; Murakami, Hiroshi; Kitajima, Sachiko; Mizokami, Masashi

    2009-03-01

    Measurements of serum concentrations of Des-gamma-carboxy Prothrombin (PIVKA-II) are widely used for diagnosing hepatocellular carcinoma (HCC). Recently, in Lumipulsef assay, it was reported that antibodies against alkaline phosphatase (ALP) derived from anti bleeding sheets led false high values of PIVKA-II in the patients with HCC resection. To improve the previous issue, newly developed Lumipulse PrestoII assay was examined. (1) The assay was reliable and positively correlated with the previous assays (Lumipulse f and Picolumi, R = 0.997 and 0.994 (n=115), respectively). (2) Eleven cases, which had false high values of PIVKA-II by the Lumipulsef assay, were examined by the PrestoII assay with excess of inactive ALP. The false high values of 10 cases were improved, but only one was still high. False reactivity of this case was stronger than other cases, more effective adsorption was required. (3) Comparing the absorbent activity of inactive ALP among 6 different kinds, we found inactive ALP with much higher adsorbent activity. When this inactive ALP was applied to assay, false high values of PIVKA-II were improved in all 11 cases. In conclusion, the PrestoII assay, which applies the inactive ALP with high activity, is reliable and useful for clinical screening. PMID:19363989

  6. Bioprecipitation of uranium from alkaline waste solutions using recombinant Deinococcus radiodurans

    Energy Technology Data Exchange (ETDEWEB)

    Kulkarni, Sayali; Ballal, Anand; Apte, Shree Kumar, E-mail: aptesk@barc.gov.in

    2013-11-15

    Highlights: • Deinococcus radiodurans was genetically engineered to overexpress alkaline phosphatase (PhoK). • Deino-PhoK bioprecipitated U efficiently over a wide range of input U concentration. • A maximal loading of 10.7 g U/g of biomass at 10 mM input U was observed. • Radioresistance and U precipitation by Deino-PhoK remained unaffected by ? radiation. • Immobilization of Deino-PhoK facilitated easy separation of precipitated U. -- Abstract: Bioremediation of uranium (U) from alkaline waste solutions remains inadequately explored. We engineered the phoK gene (encoding a novel alkaline phosphatase, PhoK) from Sphingomonas sp. for overexpression in the radioresistant bacterium Deinococcus radiodurans. The recombinant strain thus obtained (Deino-PhoK) exhibited remarkably high alkaline phosphatase activity as evidenced by zymographic and enzyme activity assays. Deino-PhoK cells could efficiently precipitate uranium over a wide range of input U concentrations. At low uranyl concentrations (1 mM), the strain precipitated >90% of uranium within 2 h while a high loading capacity of around 10.7 g U/g of dry weight of cells was achieved at 10 mM U concentration. Uranium bioprecipitation by Deino-PhoK cells was not affected in the presence of Cs and Sr, commonly present in intermediate and low level liquid radioactive waste, or after exposure to very high doses of ionizing radiation. Transmission electron micrographs revealed the extracellular nature of bioprecipitated U, while X-ray diffraction and fluorescence analysis identified the precipitated uranyl phosphate species as chernikovite. When immobilized into calcium alginate beads, Deino-PhoK cells efficiently removed uranium, which remained trapped in beads, thus accomplishing physical separation of precipitated uranyl phosphate from solutions. The data demonstrate superior ability of Deino-PhoK, over earlier reported strains, in removal of uranium from alkaline solutions and its potential use in bioremediation of nuclear and other waste.

  7. Alkaline fuel cells applications

    Science.gov (United States)

    Kordesch, Karl; Hacker, Viktor; Gsellmann, Josef; Cifrain, Martin; Faleschini, Gottfried; Enzinger, Peter; Fankhauser, Robert; Ortner, Markus; Muhr, Michael; Aronson, Robert R.

    On the world-wide automobile market technical developments are increasingly determined by the dramatic restriction on emissions as well as the regimentation of fuel consumption by legislation. Therefore there is an increasing chance of a completely new technology breakthrough if it offers new opportunities, meeting the requirements of resource preservation and emission restrictions. Fuel cell technology offers the possibility to excel in today's motive power techniques in terms of environmental compatibility, consumer's profit, costs of maintenance and efficiency. The key question is economy. This will be decided by the costs of fuel cell systems if they are to be used as power generators for future electric vehicles. The alkaline hydrogen-air fuel cell system with circulating KOH electrolyte and low-cost catalysed carbon electrodes could be a promising alternative. Based on the experiences of Kordesch [K. Kordesch, Brennstoffbatterien, Springer, Wien, 1984, ISBN 3-387-81819-7; K. Kordesch, City car with H 2-air fuel cell and lead-battery, SAE Paper No. 719015, 6th IECEC, 1971], who operated a city car hybrid vehicle on public roads for 3 years in the early 1970s, improved air electrodes plus new variations of the bipolar stack assembly developed in Graz are investigated. Primary fuel choice will be a major issue until such time as cost-effective, on-board hydrogen storage is developed. Ammonia is an interesting option. The whole system, ammonia dissociator plus alkaline fuel cell (AFC), is characterised by a simple design and high efficiency.

  8. Alkalinity and formation of zeolites in saline alkaline lakes.

    Science.gov (United States)

    Mariner, R H; Surdam, R C

    1970-11-27

    The solubility of rhyolitic glass increases with increasing alkalinity, whereas the ratio of silicon to aluminum decreases with increasing alkalinity. The strong correlation observed between alkalinity and zeolite mineralogy in saline, alkaline lakes is thought to be a function of this relationship between pH and the Si/Al ratio. It is suggested that this function is a result of the reaction between silicic glass and alkaline solution whereby (i) a gel forms, whose Si/Al ratio is controlled by the Si/Al ratio of the solution, and (ii) a zeolite forms from the gel, whose Si/Al ratio is, in turn, controlled by the composition of the gel. PMID:17834613

  9. Alkaline thermal sludge hydrolysis.

    Science.gov (United States)

    Neyens, E; Baeyens, J; Creemers, C

    2003-02-28

    The waste activated sludge (WAS) treatment of wastewater produces excess sludge which needs further treatment prior to disposal or incineration. A reduction in the amount of excess sludge produced, and the increased dewaterability of the sludge are, therefore, subject of renewed attention and research. A lot of research covers the nature of the sludge solids and associated water. An improved dewaterability requires the disruption of the sludge cell structure. Previous investigations are reviewed in the paper. Thermal hydrolysis is recognized as having the best potential to meet the objectives and acid thermal hydrolysis is most frequently used, despite its serious drawbacks (corrosion, required post-neutralization, solubilization of heavy metals and phosphates, etc.). Alkaline thermal hydrolysis has been studied to a lesser extent, and is the subject of the detailed laboratory-scale research reported in this paper. After assessing the effect of monovalent/divalent cations (respectively, K(+)/Na(+) and Ca(2+)/Mg(2+)) on the sludge dewaterability, only the use of Ca(2+) appears to offer the best solution. The lesser effects of K(+), Na(+) and Mg(2+) confirm previous experimental findings. As a result of the experimental investigations, it can be concluded that alkaline thermal hydrolysis using Ca(OH)(2) is efficient in reducing the residual sludge amounts and in improving the dewaterability. The objectives are fully met at a temperature of 100 degrees C; at a pH approximately 10 and for a 60-min reaction time, where all pathogens are moreover killed. Under these optimum conditions, the rate of mechanical dewatering increases (the capillary suction time (CST) value is decreased from approximately 34s for the initial untreated sample to approximately 22s for the hydrolyzed sludge sample) and the amount of DS to be dewatered is reduced to approximately 60% of the initial untreated amount. The DS-content of the dewatered cake will be increased from 28 (untreated) to 46%.Finally, the mass and energy balances of a wastewater treatment plant with/without advanced sludge treatment (AST) are compared. The data clearly illustrate the benefits of using an alkaline AST-step in the system. PMID:12573845

  10. BIOCHEMICAL INDICATORS OF HEPATOTOXICITY IN BLOOD SERUM OF RATS UNDER THE EFFECT OF NOVEL 4-THIAZOLIDINONE DERIVATIVES AND DOXORUBICIN AND THEIR COMPLEXES WITH POLYETHYLENEGLYCOL-CONTAINING NANOSCALE POLYMERIC CARRIER.

    Science.gov (United States)

    Kobylinska, L I; Havrylyuk, D Ya; Ryabtseva, A O; Mitina, N E; Zaichenko, O S; Lesyk, R B; Zimenkovsky, B S; Stoika, R S

    2015-01-01

    The aim of this study was to compare the effect of new synthetic 4-tiazolidinone derivatives (compounds 3882, 3288 and 3833) and doxorubicin (positive control) in free form and in their complexes with synthetic polyethyleneglycol-containing nanoscale polymeric carrier on the biochemical indicators of hepatotoxicity in blood serum of rats. The activity of enzymes considered as the markers of hepatotoxicity, as well as. the concentration of total protein, urea and creatinine were measured in blood serum of rats. It was found that after injection of investigated compounds the activities ofalanine aminotransferase, alkaline phosphatase and ?-amylase increased in comparison to control. Doxorubicin injection was accompanied by 4-fold increase in the activity of ?-glutamyltransferase, and injection ofcompound 3833 led to 2.5-fold elevation ofthe activity of this enzyme. Complexation ofthese antineoplastic derivatives with a synthetic nanocarrier lowered the activity ofthe investigated enzymes substantially if compared to the effect of these compounds infreeform. The most evident decrease was measured for ?-amylase, ?-glutamyltransferase and lactate dehydrogenase activities. The normalization of concentrations of total protein, urea and creatinine in blood serum of rats treated with complexes of the studied compounds with a polymeric carrier comparing with their introduction infreeform was also detected. Thus, the immobilization by novel polymeric carrier of anticancer drugs possessing high general toxicity in the treated organism mitigates their toxic effect, which is evident as normalization of specific biochemical indicators of the hepatodestructive effects of the anticancer drugs. PMID:26255346

  11. Alkaline fuel cells applications

    Energy Technology Data Exchange (ETDEWEB)

    Kordesch, K.; Gsellmann, J.; Cifrain, M. [Institute for Chemical Technology of Inorganic Materials, Technical University Graz (Australia); Hacker, V.; Faleschini, G.; Enzinger, P.; Fankhauser, R.; Ortner, M.; Muhr, M. [Innovative Energy Technology, Institute of High Voltage Engineering, Technical University Garz (Australia); Aronson, R.R. [Electric Auto Corporation (EAC), Fort Lauderdale, FL (United States)

    2000-03-01

    On the world-wide automobile market technical developments are increasingly determined by the dramatic restriction on emissions as well as the regimentation of fuel consumption by legislation. Therefore there is an increasing chance of a completely new technology breakthrough if it offers new opportunities, meeting the requirements of resource preservation and emission restrictions. Fuel cell technology offers the possibility to excel in today's motive power techniques in terms of environmental compatibility, consumer's profit, costs of maintenance and efficiency. The key question is economy. This will be decided by the cost of fuel cell systems if they are to be used as power generators for future electric vehicles. The alkaline hydrogen-air fuel cell system with circulating KOH electrolyte and low-cost catalysed carbon electrodes could be a promising alternative. Based on the experiences of Kordesch [K. Kordesch, Brennstoffbatterien, Springer, Wien, 1984, ISBN 3-387-81819-7; K. Kordesch, City car with H{sub 2}-air fuel cell and lead-battery, SAE Paper No. 719015, 6th IECEC, 1971], who operated a city car hybrid vehicle on public roads for 3 years in the early 1970s, improved air electrodes plus new variations of the bipolar stack assembly developed in Graz are investigated. Primary fuel choice will be a major issue until such time as cost-effective, on-board hydrogen storage is developed. Ammonia is an interesting option. The whole system, ammonia dissociator plus alkaline fuel cell (AFC), is characterised by a simple design and high efficiency. (orig.)

  12. The effect of benzene on serum hormones and the activity of some enzymes in different tissues of rats

    Directory of Open Access Journals (Sweden)

    Dere Egemen D.

    2003-01-01

    Full Text Available The effects of a 100 mg.kg -1 dose of benzene, an occupational and environmental toxicant, were investigated on serum, estradiol and testosterone concentrations as well as lactate dehydrogenase (LDH, alkaline phosphatase (ALP, alanine amino transferase (ALT, aspartate amino transferase (AST, and pyruvate kinase (PK activities in the liver and kidney of rats after 2, 4, 8, 16, 32, 64 and 72 hours. Benzene was given intraperitoneally to Rat rattus norvegicus and the control groups were injected with physiological saline. Liver tissue LDH, AST, ALT and kidney tissue LDH, ALP, AST, ALT activities were lower in the benzene treated group when compared to those in the control group (p<0.05. A tendency for an increase in the liver tissue ALP activity was observed, which was significant at 8 and 16 hours (p<0.05. There were significant increases in ALT in the liver and LDH, AST, and ALT enzyme activities in the kidney tissue at the beginning of the experiment in both groups and these activities were found to be nearly the same. Pyruvate kinase enzyme activities in rats given benzene were slightly increased in kidney tissues but lower in liver tissues. Differences between the groups tended to disappear towards the end of the experimental period. However, serum estradiol concentrations in the serum diverged significantly (p<0.05. Consequently, it was found that benzene administration led to some changes (increases then decreases in LDH, ALP, ALT, AST, and PK activity and estradiol, testosterone concentrations in different tissues of rats. Possible causes of the increases and decreases in enzyme activities and hormone levels are discussed.

  13. Evaluation of Changes of Factors Related to Liver Function in Serum of Horse by Administration of Cichorium intybus

    Directory of Open Access Journals (Sweden)

    H. Najafzadeh

    2011-02-01

    Full Text Available Chicory (Cichorium intybus is a plant that is cultured in some area of Iran, including Khozestan. All of parts of the chicory especially its leave and root have medicinal properties. It is traditionally used for treatment icterus, renal failure, gout and arthritis in human. Important side effects were not reported from this plant. Excretion substances like uric acid are clinically important in some pathological conditions such as urecemia and icterus in horse. However the pharmacological effect of chicory was not evaluated in horses. The present study was conducted for evaluation effect of leave of chicory on changes of factors related to liver function in serum of horse. In this study, 8 Arabian horses were selected. They had 10-20 years and were clinically in normal conditions. The horses were fed routine diet. The dried leave of chicory was daily added to food of horses at 0.5 g/kg for 15 days. The blood of horses was daily collected before, during and 6 days after chicory administration. The serum was isolated and uric acid, Alanine Transferase (ALT, Aspartate Transferase (AST, Alkaline Phosphatase (ALP, Lactate Dehydrogenase (LDH, conjugated and total billirubin, total protein and albumin concentrations were measured. The mean of these factors were statistically compared. Chicory consumption did not statistically change concentration of above factors. Thus, chicory dose not affect concentration of ALT, AST, ALP, LDH, conjugated and total billirubin, total protein and albumin and uric acid in serum of horse in normal condition; but it may be benefit in pathological conditions.

  14. Growth, serum biochemistry, complement activity, and liver gene expression responses of Pekin ducklings to graded levels of cultured aflatoxin B1.

    Science.gov (United States)

    Chen, X; Horn, N; Cotter, P F; Applegate, T J

    2014-08-01

    A 14-d study was conducted to evaluate the effects of cultured aflatoxin B1 (AFB1) on performance, serum biochemistry, serum natural antibody and complement activity, and hepatic gene expression parameters in Pekin ducklings. A total of 144 male Pekin ducklings were weighed, tagged, and randomly allotted to 4 dietary treatments containing 4 concentrations of AFB1 (0, 0.11, 0.14, and 0.21 mg/kg) from 0 to 14 d of age (6 cages per diet; 6 ducklings per cage). Compared with the control group, there was a 10.9, 31.7, and 47.4% (P urea N, Cl, alkaline phosphatase, and aspartate amino transferase concentrations increased linearly with increasing AFB1 (P rabbit, human type O, and horse erythrocytes, and decreased rabbit and horse agglutinins (P < 0.05). Liver peroxisome proliferator activated receptor ? (PPAR?) expression was linearly downregulated by AFB1 (P < 0.01). Results from this study indicate that for every 0.10 mg/kg increase in dietary AFB1, cumulative feed intake and BW gain decrease approximately 230 and 169 g per duckling from hatch to 14 d; and that AFB1 at very low concentrations can significantly impair liver function and gene expression, and innate immune dynamics in Pekin ducklings. PMID:24902705

  15. Enhanced Activity of Alkaline Phosphatases - Phytoplankton Response to Epilimnetic Phosphorus Depletion.

    Czech Academy of Sciences Publication Activity Database

    Vrba, Jaroslav; Komárková, Jaroslava; Vyhnálek, Vojt?ch

    1993-01-01

    Ro?. 28, ?. 6 (1993), s. 15-24. ISSN 0273-1223. [2nd International Conference on Reservoir Limnology and Water Quality. ?. Bud?jovice, 09.08.1992-14.08.1992] R&D Projects: GA AV ?R IA61706 Impact factor: 0.573, year: 1993

  16. Localization of inclusion bodies in Escherichia coli overproducing beta-lactamase or alkaline phosphatase.

    OpenAIRE

    Georgiou, G.(); Telford, J N; Shuler, M L; Wilson, D. B.

    1986-01-01

    High-level synthesis of the periplasmic protein beta-lactamase in Escherichia coli caused the formation of insoluble protein precipitates called inclusion bodies. beta-Lactamase inclusion bodies differed from those reported previously in that they appeared to be localized in the periplasmic space, not in the cytoplasm. The inclusion bodies contained mature beta-lactamase and were solubilized more easily than has been reported for cytoplasmic inclusion bodies. In contrast, overproduction of th...

  17. Zinc deficiency negatively affects alkaline phosphatase and the concentration of Ca, Mg and P in rats

    OpenAIRE

    Cho, Young-Eun; Lomeda, Ria-Ann R.; Ryu, Sang-Hoon; Sohn, Ho-Yong; Shin, Hong-In; Beattie, John H.; Kwun, In-Sook

    2007-01-01

    Zn is an essential nutrient that is required in humans and animals for many physiological functions, including immune and antioxidant function, growth, and reproduction. The present study evaluated whether Zn deficiency would negatively affect bone-related enzyme, ALP, and other bone-related minerals (Ca, P and Mg) in rats. Thirty Sprague Dawley rats were assigned to one of the three different Zn dietary groups, such as Zn adequate (ZA, 35 mg/kg), pair fed (PF, 35 mg/kg), Zn deficient (ZD, 1 ...

  18. Resveratrol Increases Bone Mineral Density and Bone Alkaline Phosphatase in Obese Men

    DEFF Research Database (Denmark)

    Ornstrup, Marie Juul; Harsløf, Torben; Kjær, Thomas Nordstrøm; Langdahl, Bente Lomholt; Pedersen, Steen Bønløkke

    2014-01-01

    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 minera...

  19. Resolvin E1-induced intestinal alkaline phosphatase promotes resolution of inflammation through LPS detoxification

    OpenAIRE

    Campbell, Eric L.; MacManus, Christopher F.; Kominsky, Douglas J; Keely, Simon; Glover, Louise E; Bowers, Brittelle E.; Scully, Melanie; Bruyninckx, Walter J; Colgan, Sean P

    2010-01-01

    Resolvin-E1 (RvE1) has been demonstrated to promote inflammatory resolution in numerous disease models. Given the importance of epithelial cells to coordination of mucosal inflammation, we hypothesized that RvE1 elicits an epithelial resolution signature. Initial studies revealed that the RvE1-receptor (ChemR23) is expressed on intestinal epithelial cells (IECs) and that microarray profiling of cells exposed to RvE1 revealed regulation of inflammatory response gene expression. Notably, RvE1 i...

  20. Phosphate solubilizing bacteria and alkaline phosphatase activity in coastal waters off Trivandrum

    Digital Repository Service at National Institute of Oceanography (India)

    Mamatha, S.S.; Gobika, A.; Janani, P.

    stream_size 22852 stream_content_type text/plain stream_name J_Coast_Environ_3_89.pdf.txt stream_source_info J_Coast_Environ_3_89.pdf.txt Content-Encoding UTF-8 Content-Type text/plain; charset=UTF-8 Phosphate Solubilizing... of phosphobacteria in the marine environment at Porto- Novo. Current Science. 39. pp. 398-399. Ahmed, N. and Shahab S. 2011. Phosphate Solubilization: Their Mechanism Genetics and Application. The Internet Journal of Microbiology. 9. Boavida, M. J. 1990. Natural...

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    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 brus...

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

    Czech Academy of Sciences Publication Activity Database

    Valdespino-Castillo, P.M.; Alcantara-Hernandez, R.J.; Alcocer, J.; Merino-Ibarra, M.; Macek, Miroslav; Falcon, L.I.

    2014-01-01

    Ro?. 90, ?. 2 (2014), s. 504-519. ISSN 0168-6496 Institutional support: RVO:60077344 Keywords : dissolved organic phosphorus utilization * extracellular enzymes * microbial functional diversity Subject RIV: EE - Microbiology, Virology Impact factor: 3.568, year: 2014

  3. Method of metabolic staining of the ERM - alkaline phosphatase and NADH diaphorase.

    Czech Academy of Sciences Publication Activity Database

    Vosátka, Miroslav

    Uppsala : Department of Microbiology, Swedish University of Agricultural Sciences, 1998 - (Kling, M.), s. 19-20 [ICOM 2 Workshop. Uppsala (SE), 01.07.1998-04.07.1998] R&D Projects: GA AV ?R KSK2017602 Subject RIV: EF - Botanics

  4. Creatine kinase serum activity in feline hyperthyroidism / Atividade sérica da creatinina quinase no hipertireoidismo felino

    Scientific Electronic Library Online (English)

    Mauro José Lahm, Cardoso; Fabiano Séllos, Costa; Luciane, Holsback; Thais Helena Constantino, Patelli; Maíra, Melussi; Ademir, Zacarias Júnior; Rafael, Fagnani.

    2014-12-01

    Full Text Available O hipertireoidismo é uma doença endócrina comum em gatos, resultante da excessiva secreção dos hormônios tireoidianos (tiroxina-T4 e triiodotironina-T3), possui caráter multissistemico e provoca várias alterações nos parametros bioquímicos. A creatina quinase (CK) é uma enzima que tem sua atividade [...] sérica aumentada em diversas enfermidades e, na medicina veterinária, é principalmente utilizada para avaliar lesões no músculo esquelético, músculo cardíaco e tecido hepático. O objetivo deste estudo foi determinar a ocorrência da atividade sérica da CK em gatos com hipertireoidismo e suas associações com alterações clínico-patológicas e com a tiroxina total (TT4) e livre (FT4). A atividade sérica da CK foi avaliada em 19 gatos com hipertireodismo espontâneo, com TT4 variando de 34,88-294,98nmol L-1 e FT4 variando de 2,83-52,9nmol L-1. Sua atividade estava elevada em 47,3% dos gatos (?250U L-1) e apresentou diferença estatística (P Abstract in english The aim of this study is to determine the occurrence of creatine kinase (CK) serum activity in cats with hyperthyroidism and its associations with clinical-pathological alterations and with total (TT4) and free (FT4) thyroxin. CK serum activity was evaluated in 19 cats with spontaneous hyperthyroidi [...] sm, with TT4 ranging from 34.88-294.98nmol L-1 and FT4 ranging from 2.83-52.9pmol L-1 and also by serum biochemical analysis, including the activity of CK, alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), cholesterol and bile acids. The reference value for CK considered in this study was of 110-250U L-1. CK serum activity was evaluated in 19 cats with spontaneous hyperthyroidism, with TT4 ranging from 34.88-294.98nmol L-1 and FT4 ranging from 2.83-52. 9pmol L-1. Its activity CK was elevated in 47.3% of cats (?250U L-1) and presented statistical difference (P

  5. Interaction of alkali and alkaline earth ions with Ochratoxin A

    International Nuclear Information System (INIS)

    The effect of alkali and alkaline earth ions on the chemical equilibrium of mono- and dianionic forms of the mycotoxin Ochratoxin A (OTA) and their bonding onto the surface of Bovine Serum Albumin (BSA) have been investigated by fluorescence spectroscopy and fluorescence polarization techniques. Our results show that alkali metal ions shift the chemical equilibrium towards formation of dianionic form of OTA. Furthermore, the alkaline earth ions can compete with BSA for binding to OTA when these ions are present in millimolar concentrations. Our data also highlight the possibility that the “free” fraction of OTA (not bound onto the surface of albumin) or at least a part of it is present in cation-bound form in body fluids. These observations are supported by stability constants and quantum-chemical calculations. Among the studied alkaline metal ions magnesium showed the highest affinity towards OTA under physiological conditions. Further research is required to analyze the potential significance of Mg2+–OTA complex in cellular uptake and/or elimination of the toxin in the human body. - Highlights: ? Fluorescence spectroscopy reveals cation–Ochratoxin A (OTA) interactions. ? Alkali ions shift the equilibrium of OTA to formation of a dianionic structure. ? Alkaline earth ions directly bind to OTA in the order: Mg2+, Ca2+, Ba2+. ? Quantum chemical calculations and logK values support our experimental data.

  6. Serum osteoprotegerin as a screening tool for coronary artery calcification score in diabetic pre-dialysis patients

    International Nuclear Information System (INIS)

    Although cardiovascular disease is a principal cause of death in patients with chronic kidney disease (CKD), it is often asymptomatic in diabetic patients. The coronary artery calcification score (CACS) measured by multidetector computed tomography (MDCT) is useful for screening ischemic heart disease in the general population. We investigated which clinical parameters predict high CACS in predialysis diabetic nephropathy (DN). Participants were 85 patients with DN. Nobody had any history of coronary angioplasty or coronary bypass surgery. We measured blood counts, blood chemistry, bone alkaline phosphatase, intact-parathyroid hormone (PTH), interleukin-6, osteoprotegerin (OPG), hemoglobin A1c, 25-hydroxyvitamin D (25(OH)D) and fetuin-A. CACS and bone mineral density (BMD) were measured by a single 16-slice MDCT and Dual Energy X-ray Absorptiometry (DEXA), respectively. The median value of CACS equaled 256 Agatston units (range 0-4494 units). Stepwise increase in CACS with CKD stage progression was observed (p200 was 80%, when the cut-off value was 1.2 ng/mL. In conclusion, CACS increased with CKD stage progression in predialysis DN patients. Serum OPG was positively associated with high CACS and can be a useful screening tool for severe coronary calcification, whereas no association between fetuin-A and CACS was found. (author)

  7. The effect of Stevia rebaudiana on serum omentin and visfatin level in STZ-induced diabetic rats.

    Science.gov (United States)

    Akbarzadeh, Samad; Eskandari, Fatemeh; Tangestani, Hadis; Bagherinejad, Somaieh Tangerami; Bargahi, Afshar; Bazzi, Parviz; Daneshi, Adel; Sahrapoor, Azam; O'Connor, William J; Rahbar, Ali Reza

    2015-03-01

    Recently the role of adipocytokines in relationship to incidence of diabetes has been demonstrated. One of the medicinal plants that are used in the treatment of diabetes is stevia. This study investigates the effect of stevia on serum omentin and visfatin levels as novel adipocytokines in diabetic induced rats to find potential mechanisms for the anti hyperglycemic effect of stevia. Forty male wistar rats weighing 180-250 g were induced with diabetes by intraperitoneal injection of streptozotocin (STZ). The animals were divided into 5 groups of 8. Rats in group 1 (non-diabetic control) and group 2 (diabetic control) were treated with distilled water, and the rats in the treated groups, group 3 (T250), group 4 (T500), and group 5 (T750) were treated with stevia, gavaged every day at 9 a.m. in doses of 250, 500, and 750 mg/kg, respectively. At the end of the study significant reductions in fasting blood sugar (FBS), the homeostasis model assessment insulin resistance (HOMA-IR), triglyceride (TG), alkaline phosphatase (ALP), and Omentin level were found in groups 3 and 4 in comparison with group 2. Pancreatic histopathology slides demonstrated that stevia extract did not induce any increase in the number of ?-cells. The conclusion is that prescription of stevia in the doses of 250 and 500 mg/kg/d decreases the omentin level indirectly via activating insulin sensitivity and lowering blood glucose in STZ-induced diabetic rats. PMID:24689449

  8. [Changes of serum enzymes, lactate and hemoglobin concentrations in the blood of young trotting horses due to training exertion].

    Science.gov (United States)

    Krzywanek, H; Mohr, E; Mill, J; Scharpenack, M

    1996-08-01

    Until the age of about 2 years, trotters normally grow up on pasture without any kind of training. In the stud farm Lindenhof (Templin, Germany), however, these first 2 years are used for a special fitness training for the young animals: 2-3 times a week, a group of the yearlings is forced to run a distance of about 1700 m on a track at an average speed of up to 10 m/s. Until now, little was known about changes of blood parameters which may occur during such special exercise. This study therefore investigated the activity of selected serum enzymes (aspartate-amino-transferase (AST), alanine-amino-transferase (ALT), gamma-glutamyl-transferase (gamma-GT), lactic dehydrogenase (LDH), alkaline phosphatase (AP), creatine kinase (CK)) and the variations of hematocrit, hemoglobin, lactate, protein, and urea concentration before and after exercise. Except for activity of AP and CK and concentration of urea, all parameters showed a distinct increase after exercise. In particular, the rise in lactic-acid concentration with values up to 23.08 mmol/l was remarkable, however, none of the parameters reached a pathological level. It is therefore concluded that exercise for young trotters over a medium distance-even at high speed-does not cause any injury of myocardium, skeletal muscles or liver cells. PMID:9005684

  9. Titania nanotube delivery fetal bovine serum for enhancing MC3T3-E1 activity and osteogenic gene expression.

    Science.gov (United States)

    Peng, Jing; Zhang, Xinming; Li, Zhaoyang; Liu, Yunde; Yang, Xianjin

    2015-11-01

    Titania nanotube (TNT) delivery of fetal bovine serum (FBS) was conducted on titanium (Ti) to enhance bone tissue repair. Scanning electron microscopy (SEM) and energy dispersive spectrometer (EDS) showed FBS increased the tube wall thickness and decreased the tube diameter. Attenuated total reflectance Fourier transform infrared further confirmed that FBS completely covered the TNT and changed the surface composition. Water contact angle tests showed TNT/FBS possessed hydrophilic properties. Compared to original Ti, the TNT/FBS group had more attached osteoblasts after 2h and enhanced filopodia growth at 0.5h. Significantly, more osteoblasts were also observed on TNT/FBS after 7d culturing. FBS was released steadily from TNT; about 70% of FBS had been released at 3d and 90% at 5d, as shown by the bicinchoninic acid method. TNT/FBS also enhanced subsequent osteoblast differentiation and gene expression; the quantum real-time polymerase chain reaction test showed that TNT/FBS up-regulated alkaline phosphatase and osteocalcin gene expression at 7d and 14d. Therefore, TNT/FBS delivered sustained in situ nutrition and enhanced osteoblast activity and osteogenic gene expression. PMID:26249612

  10. Identification of PTP? as an autophagic phosphatase

    OpenAIRE

    Martin, Katie R; Xu, Yong; Looyenga, Brendan D.; Davis, Ryan J.; Wu, Chia-Lun; Tremblay, Michel L; Xu, H. Eric; MacKeigan, Jeffrey P.

    2011-01-01

    Macroautophagy is a dynamic process whereby portions of the cytosol are encapsulated in double-membrane vesicles and delivered to the lysosome for degradation. Phosphatidylinositol-3-phosphate (PtdIns3P) is concentrated on autophagic vesicles and recruits effector proteins that are crucial for this process. The production of PtdIns3P by the class III phosphatidylinositol 3-kinase Vps34, has been well established; however, protein phosphatases that antagonize this early step in autophagy remai...

  11. Leishmanial phosphatase hydrolyzes phosphoproteins and inositol phosphates

    International Nuclear Information System (INIS)

    An extensively purified preparation of the predominant, tartrate-resistant acid phosphatase (ACP) from the external surface of Leishmania donovani promastigotes form catalyzes the dephosphorylation of several phosphoproteins; these include: pyruvate kinase, phosphorylase kinase and histones. However, the protein phosphatase activity of ACP is very low compared with that of other protein phosphates known to be involved in regulating various metabolic pathways. 32P-labelled inositoltriphosphate (IP3), a well-established second messenger derived from phosphatidylinositol-4,5-diphosphate (PIP2), was a substrate for the leishmanial acid phosphatase; incubation of the IP3 preparation with 13.2 milliunits (1 unit equals 1 ?mol 4-methylumbelliferyl phosphate (MUP) cleaved per min at pH 5.5) of ACP at pH 5.5 for 4 hr resulted in hydrolysis of 75% of the radiolabelled substrate resulting in a mixture of inositoldiphosphate and inositolmonophosphate. In addition PIP2 was hydrolyzed rapidly by ACP at pH 5.5 (V/sub max/, 71 units/mg protein; k/sub m/, 4.16 ?M). In contrast, to MUP which is hydrolzyed most rapidly at pH 5.5, PIP2 hydrolysis was optimal at pH 6.8. These observations raise the possibility that ACP could play a role in the host-phagocyte interaction by degrading the precursor of the second messenger, PIP2 or the second messenger itself, IP3

  12. Identification of PTP? as an autophagic phosphatase

    Science.gov (United States)

    Martin, Katie R.; Xu, Yong; Looyenga, Brendan D.; Davis, Ryan J.; Wu, Chia-Lun; Tremblay, Michel L.; Xu, H. Eric; MacKeigan, Jeffrey P.

    2011-01-01

    Macroautophagy is a dynamic process whereby portions of the cytosol are encapsulated in double-membrane vesicles and delivered to the lysosome for degradation. Phosphatidylinositol-3-phosphate (PtdIns3P) is concentrated on autophagic vesicles and recruits effector proteins that are crucial for this process. The production of PtdIns3P by the class III phosphatidylinositol 3-kinase Vps34, has been well established; however, protein phosphatases that antagonize this early step in autophagy remain to be identified. To identify such enzymes, we screened human phosphatase genes by RNA interference and found that loss of PTP?, a dual-domain protein tyrosine phosphatase (PTP), increases levels of cellular PtdIns3P. The abundant PtdIns3P-positive vesicles conferred by loss of PTP? strikingly phenocopied those observed in cells starved of amino acids. Accordingly, we discovered that loss of PTP? hyperactivates both constitutive and induced autophagy. Finally, we found that PTP? localizes to PtdIns3P-positive membranes in cells, and this vesicular localization is enhanced during autophagy. We therefore describe a novel role for PTP? and provide insight into the regulation of autophagy. Mechanistic knowledge of this process is crucial for understanding and targeting therapies for several human diseases, including cancer and Alzheimer's disease, in which abnormal autophagy might be pathological. PMID:21303930

  13. Synthesis of functionalized fluorescent gold nanoclusters for acid phosphatase sensing

    Science.gov (United States)

    Sun, Jian; Yang, Fan; Yang, Xiurong

    2015-10-01

    A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by introducing an alkaline aqueous solution of MUA into the GSH-Au+ complexes or AuNC@GSH solution. Subsequently, a reliable AuNC@GSH/MUA-based real-time assay of acid phosphatase (ACP) is established for the first time, inspired by the selective coordination of Fe3+ with surface ligands of AuNCs, the higher binding affinity between the pyrophosphate ion (PPi) and Fe3+, and the hydrolysis of PPi into orthophosphate by ACP. Our fluorescent chemosensor can also be applied to assay ACP in a real biological sample and, furthermore, to screen the inhibitor of ACP. This report paves a new avenue for synthesizing AuNCs based on either the bottom-up reduction or top-down etching method, establishing real-time fluorescence assays for ACP by means of PPi as the substrate, and further exploring the sensing applications of fluorescent AuNCs.A novel and convenient one-pot but two-step synthesis of fluorescent gold nanoclusters, incorporating glutathione (GSH) and 11-mercaptoundecanoic acid (MUA) as the functionalized ligands (i.e. AuNCs@GSH/MUA), is demonstrated. Herein, the mixing of HAuCl4 and GSH in aqueous solution results in the immediate formation of non-fluorescent GSH-Au+ complexes, and then a class of ~2.6 nm GSH-coated AuNCs (AuNCs@GSH) with mild orange-yellow fluorescence after several days. Interestingly, the intense orange-red emitting ~1.7 nm AuNCs@GSH/MUA can be synthesized within seconds by introducing an alkaline aqueous solution of MUA into the GSH-Au+ complexes or AuNC@GSH solution. Subsequently, a reliable AuNC@GSH/MUA-based real-time assay of acid phosphatase (ACP) is established for the first time, inspired by the selective coordination of Fe3+ with surface ligands of AuNCs, the higher binding affinity between the pyrophosphate ion (PPi) and Fe3+, and the hydrolysis of PPi into orthophosphate by ACP. Our fluorescent chemosensor can also be applied to assay ACP in a real biological sample and, furthermore, to screen the inhibitor of ACP. This report paves a new avenue for synthesizing AuNCs based on either the bottom-up reduction or top-down etching method, establishing real-time fluorescence assays for ACP by means of PPi as the substrate, and further exploring the sensing applications of fluorescent AuNCs. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr04826e

  14. Bismuth citrate in the quantification of inorganic phosphate and its utility in the determination of membrane-bound phosphatases.

    Science.gov (United States)

    Cariani, L; Thomas, L; Brito, J; del Castillo, J R

    2004-01-01

    This paper describes a rapid and sensitive method to determine inorganic phosphate, even in the presence of labile organic phosphate compounds and large quantities of proteins. The method eliminates the use of sodium arsenite, a highly toxic compound, substituting bismuth citrate for it to stabilize the phosphomolybdic acid complex formed during the interaction of inorganic phosphate and molybdate reduced by ascorbic acid. This method has also been adapted to microplates and has been used to determine the activities of Na/K ATPase and alkaline phosphatase of intestinal basolateral and luminal plasma membranes. PMID:14654048

  15. Passive immunity transfer and serum constituents of crossbred calves / Transferência de imunidade passiva e constituintes séricos de bezerros mestiços

    Scientific Electronic Library Online (English)

    Thaís G., Rocha; Ricardo P., Nociti; Alexandre A.M., Sampaio; José Jurandir, Fagliari.

    2012-06-01

    Full Text Available A avaliação da transferência de imunidade passiva (TIP) é uma ferramenta essencial para manutenção de bezerros saudáveis nos primeiros meses de vida. Uma vez que há influência do número de lactações e da raça de vacas nos teores de imunoglobulinas do colostro, o presente estudo foi conduzido com o o [...] bjetivo de avaliar a TIP de vacas Canchim primíparas e pluríparas aos seus bezerros. Amostras de sangue dos bezerros foram coletadas antes da ingestão de colostro e 1, 2, 7, 15 e 30 dias após o nascimento e amostras de colostro das vacas foram coletadas imediatamente após o parto. As atividades de gamaglutamiltransferase (GGT), fosfatase alcalina (ALP) e as concentrações de proteína total, albumina, globulinas, imunoglobulina A (IgA) e imunoglobulina G (IgG), cálcio total e ionizado, fósforo, magnésio, sódio e potássio foram avaliadas no soro dos bezerros e as atividades de GGT e ALP e as concentrações de proteína total, IgA e IgG foram avaliadas no soro colostral. A concentração de imunoglobulinas foi avaliada por meio de eletroforese em gel de poliacrilamida. As avaliações bioquímicas do soro sanguíneo dos bezerros revelaram aumento nas atividades das enzimas gamaglutamiltransferase e fosfatase alcalina e nos teores de proteína total, globulinas, imunoglobulina A e imunoglobulina G após a ingestão do colostro. Apenas os teores de proteína total e imunoglobulina G de cadeia leve no soro colostral foram influenciados pelo número de lactações das vacas. Os teores de fósforo e magnésio aumentaram após a ingestão de colostro, enquanto as concentrações de sódio e potássio oscilaram no decorrer do período experimental. A TIP foi influenciada pelo número de lactações das vacas, no entanto mostrou-se eficiente em ambos os grupos. Abstract in english Passive immunity transfer (PIT) evaluation is an essential tool for the maintenance of healthy calves during the first months of life. Since lactation number and breed have been proven to influence immunoglobulin levels in colostrum, the aim of this study was to evaluate PIT from primiparous and mul [...] tiparous Canchim cows to their calves. Blood samples were collected from the calves before colostrum intake and 1, 2, 7, 15 and 30 days thereafter, while colostrum samples from the cows were taken immediately after parturition. Activities of gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), and concentrations of total protein, albumin, globulins, immunoglobulin A (IgA), immunoglobulin G (IgG), total and ionized calcium, inorganic phosphorus, magnesium, sodium and potassium were evaluated in calves' serum and activities of GGT and ALP and concentrations of total protein, IgA and IgG were assessed in cow's colostrum whey. Immunoglobulins concentrations were evaluated by electrophoresis in polyacrylamide gels. Serum biochemistry evaluations revealed an increase in gamma-glutamyl transferase and alkaline phosphatase activities and in total protein, globulins, immunoglobulin A and immunoglobulin G levels in calves' serum after colostrum intake. Only total protein and light chain immunoglobulin G levels in colostrum whey were affected by the cows' lactation number. Phosphorus and magnesium levels in blood serum increased after colostrum intake, while sodium and potassium levels oscillated in the experimental period. PIT was influenced by the cows' lactation number but was efficient in both groups.

  16. Human prostatic acid phosphatase: purification, characterization, and optimization of conditions for radioimmunoassay

    International Nuclear Information System (INIS)

    Prostatic acid phosphatase was isolated from benign hypertrophic prostate tissue by ammonium sulfate precipitation and affinity chromatography procedures. The purified enzyme was characterized by two-dimensional gel electrophoresis and shown to have a cluster of protein spots with an apparent molecular weight of 48000 at pI 5.9 to 6.3 in 9 mol/l urea. The specific activity of the purified enzyme was 723 and 659 U/mg protein with ?-naphthyl phosphate at 300C and para-nitrophenyl phosphate at 370C respectively. An antibody to the purified enzyme was raised in rabbits and used in a radioimmunoassay (RIA). The use of a phosphate buffer, pH 6.6, and iodination of prostatic acid phosphatase (PAP) by the Bolton-Hunter procedure improved the precision of the assay when compared to RIA's using a phosphate buffer, pH 7.0 or 7.3, or PAP iodinated by a chloramine-T procedure. The former RIA displaced 50% of the tracer at 2 ?g of enzyme per liter of serum. The between-run coefficient of variation for 11 assays ranged from 3.9-7.7% with serum at 1.3 to 5.6 ?g PAP/l. (Auth.)

  17. Enzyme kinetic characterization of protein tyrosine phosphatases

    DEFF Research Database (Denmark)

    Peters, Günther H.j.; Branner, S.; Møller, K. B.; Andersen, J.N.; Møller, N.P.H.

    2003-01-01

    Protein tyrosine phosphatases (PTPs) play a central role in cellular signaling processes, resulting in an increased interest in modulating the activities of PTPs. We therefore decided to undertake a detailed enzyme kinetic evaluation of various transmembrane and cytosolic PTPs (PTPalpha, PTPbeta, PTPis an element of, CD45, LAR, PTP1B and SHP-1), using pNPP as substrate. Most noticeable is the increase in the turnover number for PTPbeta with increasing pH and the weak pH-dependence of the turnove...

  18. RECLAMATION OF ALKALINE ASH PILES

    Science.gov (United States)

    The objective of the study was to develop methods for reclaiming ash disposal piles for the ultimate use as agricultural or forest lands. The ashes studied were strongly alkaline and contained considerable amounts of salts and toxic boron. The ashes were produced from burning bit...

  19. Preventing Effects of Wheat Germ Oil on Sex Hormones, Liver Enzymes, Lipids and Proteins in Rat Serum Following Treatment with p-Nonylphenol

    Directory of Open Access Journals (Sweden)

    M. Soleimani Mehranjani

    2007-01-01

    Full Text Available The aim of this study is to investigate the preventing effect of wheat germ oil as a rich source of vitamin E on the serum biochemical factors in the rats exposed to para-nonylphenol. Four groups (n = 6 of male Wistar rats was orally given para-nonylphenol (200 mg kgG-1dayG-1 for 70 days. After treatment the blood samples were obtained and LH, FSH, testosterone, progesterone and estrogen, transaminases, phosphatases, LDH, triglyceride, cholesterol, HDL-cholestrol and protein analysis were carried out. Para-nonylphenol caused a significant increase in LH level where other hormones remained unchanged. In addition a significant reduction was found in AST, ALT and LDH level while alkaline phosphatase increased significantly following treatment with para-nonylphenol. There was no change in the level of lipids and proteins in this study with respect to para-nonylphenol treatment. Co-administration of wheat germ oil with para-nonylphenol compensated the imbalance of the LH, AST, ALT and LDH caused by para-nonylphenol to the control level. However treatment of the rats with only wheat germ oil caused a significant reduction in the level of the hormones except for progesterone. Administration of wheat germ oil in any case caused a reduction in triglyceride level. Co-administration of wheat germ oil with para-nonylphenol eliminated the effect of para-nonylphenol on LH hormone as well as enzymes. It seems that daily used of wheat germ oil may have some benefits to the para-nohnylphenol exposure, however more clinical studies are needed to find more information.

  20. Raloxifene preserves phenytoin and sodium valproate induced bone loss by modulating serum estradiol and TGF-?3 content in bone of female mice.

    Science.gov (United States)

    Anwar, Md Jamir; Radhakrishna, K V; Sharma, Abhay; Vohora, Divya

    2014-10-01

    Antiepileptic drugs (AEDs)-induced adverse consequences on bone are now well recognized. Despite this, there is limited data on the effect of anti-osteoporotic therapies on AEDs-induced bone loss. We hypothesize that estrogen deprivation following phenytoin (PHT) and sodium valproate (SVP) therapy could lead to adverse bony effects. Both PHT and SVP inhibit human aromatase enzyme and stimulate microsomal catabolism of oestrogens. Estrogen deficiency states are known to reduce the deposition of transforming growth factor-? (TGF-?3), a bone matrix protein, having anti-osteoclastic property. Thus, an attempt was made to investigate the effect of raloxifene, a selective oestrogen receptor modulator, in comparison with calcium and vitamin D3 (CVD) supplementation, on PHT and SVP-induced alterations in bone in mice and to unravel the role of estradiol and TGF-?3 in mediation of bony effects by either AEDs or raloxifene. Further, the effect of raloxifene on seizures and on the antiepileptic efficacy of PHT and SVP was investigated. Swiss strains of female mice were treated with PHT (35 mg/kg, p.o.) and SVP (300 mg/kg, p.o.) for 120 days to induce bone loss as evidenced by reduced bone mineral density (BMD) and altered bone turnover markers (BTMs) in lumbar bones (alkaline phosphatase, tartarate resistant acid phosphatase, hydroxyproline) and urine (calcium). The bone loss was accompanied by reduced serum estradiol levels and bone TGF-?3 content. Preventive and therapeutic treatment with raloxifene ameliorated bony alterations and was more effective than CVD. It also significantly restored estradiol and TGF-?3 levels. Deprived estrogen levels (that in turn reduced lumbar TGF-?3 content) following PHT and SVP, thus, might represent one of the various mechanisms of AEDs-induced bone loss. Raloxifene preserved the bony changes without interfering with antiepileptic efficacy of these drugs, and hence raloxifene could be a potential therapeutic option in the management of PHT and SVP-induced bone disease if clinically approved. PMID:24880111

  1. Alkaline resistant ceramics; Alkalimotstaandskraftiga keramer

    Energy Technology Data Exchange (ETDEWEB)

    Westberg, Stig-Bjoern [Vattenfall Utveckling AB, Aelvkarleby (Sweden)

    2001-02-01

    Despite durability in several environments, ceramics and refractories can not endure alkaline environments at high temperature. An example of such an environment is when burning biofuel in modern heat and power plants in which the demand for increasing efficiency results in higher combustion temperatures and content of alkaline substances in the flue gas. Some experiences of these environments has been gained from such vastly different equipment as regenerator chambers in the glass industry and MHD-generators. The grains of a ceramic material are usually bonded together by a glassy phase which despite it frequently being a minor constituent render the materials properties and limits its use at elevated temperature. The damage is usually caused by alkaline containing low-melting phases and the decrease of the viscosity of the bonding glass phase which is caused by the alkaline. The surfaces which are exposed to the flue gas in a modern power plant are not only exposed to the high temperature but also a corroding and eroding, particle containing, gas flow of high velocity. The use of conventional refractory products is limited to 1300-1350 deg C. Higher strength and fracture toughness as well as durability against gases, slag and melts at temperatures exceeding 1700 deg C are expected of the materials of the future. Continuous transport of corrosive compounds to the surface and corrosion products from the surface as well as a suitable environment for the corrosion to occur in are prerequisites for extensive corrosion to come about. The highest corrosion rate is therefore found in a temperature interval between the dew point and the melting point of the alkaline-constituent containing compound. It is therefore important that the corrosion resistance is sufficient in the environment in which alkaline containing melts or slag may appear. In environments such as these, even under normal circumstances durable ceramics, such as alumina and silicon carbide, are attacked. Furthermore, the durability of the silicon carbide is reduced already by small amount of alkaline components in the flue gas. This report is an effort to identify areas for continued research activities. The work is primarily based on conclusions drawn from published articles. The areas in which further studied are most needed are: description of the corroding environment, studies of the mechanism of corrosion and evaluation and development of bonding systems or sintering methods. Two categories of material which can be of special interest due to the inertness of the crystals are different types of mullite and spinel.

  2. Serum creatine kinase elevation associated with olanzapine treatment

    OpenAIRE

    Punukollu, Bhaskar; Rutherford, Helen

    2008-01-01

    On 2 May 2008, a 25-year-old male patient on olanzapine 15 mg developed mild central chest pain, and blood tests revealed a high creatine kinase (CK) value at 1016 iu/l. Troponin, CK-MB, CK:MB ratio, full blood count (FBC), urea and electrolytes (U&E), C reactive protein (CRP) and glucose were all normal. Liver enzymes were marginally raised: alanine aminotransferase (ALT) 91 iu/l, ?-glutamyl transferase (GGT) 46 iu/l, alkaline phosphatase (ALP) 137 iu/l. The ECG was normal and the chest pain...

  3. Isolation and Characterization of Phosphatase Enzyme from the Freshwater Macroalga Cladophora glomerata Kützing (Chlorophyta

    Directory of Open Access Journals (Sweden)

    A.M. El-Shahed

    2006-01-01

    Full Text Available Acid phosphatase enzyme (acPase has been isolated from Cladophora glomerata Kützing and its kinetic properties were investigated. The optimum pH for activity of both the two isolated fractions; the cell wall-bound and the secreted acPases was found to be 4.5. Both fractions exhibited another peak of activity at 9.5 which was attributed to an alkaline phosphatase. The optimum protein concentration that resulted in maximal enzyme activities for both fractions was 8 mg mL-1. The Km value determined with p-nitrophenyl phosphate (pNPP was 24 and 14 mM for the cell wall-bound and the secreted acPases, respectively. The enzyme was inhibited by phosphate in the form of KH2PO4, molybdate in the form of K2MoO4, zinc in the form of ZnCl2.4H2O and iodine as KI. Zinc ions were the most potent inhibitor among all the tested inhibitors (max. % inhibition was 96 and 80% for the cell wall- bound and the secreted acPases, respectively. Molybdate had the least inhibitory effects (72 and 50% on the cell wall- bound and the secreted acPases respectively at 40 mM concentration.

  4. Comparison of enzyme-linked immunosorbent assay and radioimmunoassay for prostate-specific acid phosphatase in prostatic disease

    International Nuclear Information System (INIS)

    Results of an enzyme-linked immunosorbent assay (ELISA) are compared with those of a standard radioimmunoassay (RIA) for detection and quantitation of prostate-specific acid phosphatase (EC 3.1.3.2) in serum. Control subjects, patients with benign prostatic hyperplasia, and patients in all four clinical stages of prostatic adenocarcinoma were tested. The upper limit of normal (95%of the population) by the ELISA was 2.0 ?g/L, and by the RIA was 2.2 ?g/L. In prostatic a denocarcinoma stage I (not detectable by digital rectal examination), ELISA was slightly more sensitive than RIA, but sensitivity was still relatively low (20%). As tumor mass increased (stages II through IV), the frequency of increased concentrations of prostatic acid phosphatase in serum also increased. We confirmed this increase in circulating enzyme in some cases of benign prostatic hyperplasia and suggest that this finding is related to either acinar cytolysis or an increase in acini size and number. Although prostate-specific acid phosphatase is not a cancer-specific enzyme, we conclude that its measurement may be of considerable value in monitoring prostatic disease

  5. Comparison of enzyme-linked immunosorbent assay and radioimmunoassay for prostate-specific acid phosphatase in prostatic disease

    Energy Technology Data Exchange (ETDEWEB)

    Griffiths, J.; Rippe, D.F.; Panfili, P.R.

    1982-01-01

    Results of an enzyme-linked immunosorbent assay (ELISA) are compared with those of a standard radioimmunoassay (RIA) for detection and quantitation of prostate-specific acid phosphatase (EC 3.1.3.2) in serum. Control subjects, patients with benign prostatic hyperplasia, and patients in all four clinical stages of prostatic adenocarcinoma were tested. The upper limit of normal (95%of the population) by the ELISA was 2.0 ..mu..g/L, and by the RIA was 2.2 ..mu..g/L. In prostatic a denocarcinoma stage I (not detectable by digital rectal examination), ELISA was slightly more sensitive than RIA, but sensitivity was still relatively low (20%). As tumor mass increased (stages II through IV), the frequency of increased concentrations of prostatic acid phosphatase in serum also increased. We confirmed this increase in circulating enzyme in some cases of benign prostatic hyperplasia and suggest that this finding is related to either acinar cytolysis or an increase in acini size and number. Although prostate-specific acid phosphatase is not a cancer-specific enzyme, we conclude that its measurement may be of considerable value in monitoring prostatic disease.

  6. Nucleotide sequences encoding a thermostable alkaline protease

    Science.gov (United States)

    Wilson, David B. (Ithaca, NY); Lao, Guifang (Bethesda, MD)

    1998-01-01

    Nucleotide sequences, derived from a thermophilic actinomycete microorganism, which encode a thermostable alkaline protease are disclosed. Also disclosed are variants of the nucleotide sequences which encode a polypeptide having thermostable alkaline proteolytic activity. Recombinant thermostable alkaline protease or recombinant polypeptide may be obtained by culturing in a medium a host cell genetically engineered to contain and express a nucleotide sequence according to the present invention, and recovering the recombinant thermostable alkaline protease or recombinant polypeptide from the culture medium.

  7. Nucleotide sequences encoding a thermostable alkaline protease

    Energy Technology Data Exchange (ETDEWEB)

    Wilson, D.B.; Lao, G.

    1998-01-06

    Nucleotide sequences, derived from a thermophilic actinomycete microorganism, which encode a thermostable alkaline protease are disclosed. Also disclosed are variants of the nucleotide sequences which encode a polypeptide having thermostable alkaline proteolytic activity. Recombinant thermostable alkaline protease or recombinant polypeptide may be obtained by culturing in a medium a host cell genetically engineered to contain and express a nucleotide sequence according to the present invention, and recovering the recombinant thermostable alkaline protease or recombinant polypeptide from the culture medium. 3 figs.

  8. Milk-derived proteins and minerals alter serum osteocalcin in prepubertal boys after 7 days

    DEFF Research Database (Denmark)

    Mark, A.B.; Hoppe, Camilla

    2010-01-01

    We have previously shown that at equal protein content, milk, but not meat, decreased bone turnover in boys. This suggested that milk-derived components are important for bone metabolism. In the present study, we hypothesized that milk-derived proteins (whey and casein) affect bone turnover during growth depending on the content of milk minerals (calcium and phosphorus). This was a randomized, parallel, double-blind study. Eight-year-old boys (n = 57) received 1 of 4 milk drinks: whey protein with low or high content of minerals, or casein protein with low or high content of minerals. The amount of whey and casein was identical to their content in 1.5 L of milk. We measured serum osteocalcin (sOC), bone-specific alkaline phosphatase, and C-terminal telopeptides of type 1 collagen (immunoassay) and estimated dietary intake (3-day weighed food record) at baseline and after 7 days. Only sOC was significantly affected by the treatments (P <.05). There was a significant interaction between milk-derived proteins and minerals with regard to sOC (P = .01). The intake of milk drinks containing whey increased sOC at the low content of minerals, whereas it decreased sOC at the high content of minerals (P <.05). In contrast, milk drinks containing casein increased sOC both at the low and at the high contents of minerals. In conclusion, whey and casein (corresponding to their content in 1.5 L of milk) differently affect sOC in 8-year-old boys depending on the content of milk minerals, but do not seem to affect other markers for bone turnover.

  9. Acid Phosphatase Role in Chickpea/Maize Intercropping

    OpenAIRE

    Li, S M; Li, L.(Institute of High Energy Physics, Chinese Academy of Sciences, Beijing, China; Department of Modern Physics, University of Science and Technology of China, Hefei, Anhui, China; Department of Physics, Nanjing University, Nanjing, Jiangsu, China; School of Physics, Shandong University, Jinan, Shandong, China; Physics Department, Shanghai Jiao Tong University, Shanghai, China); Zhang, F. S.; Tang, C.

    2004-01-01

    • Background and aims Organic P comprises 30–80 % of the total P in most agricultural soils. It has been proven that chickpea facilitates P uptake from an organic P source by intercropped wheat. In this study, acid phosphatase excreted from chickpea roots is quantified and the contribution of acid phosphatase to the facilitation of P uptake by intercropped maize receiving phytate is examined.

  10. Atypical Protein Phosphatases: Emerging Players in Cellular Signaling

    Directory of Open Access Journals (Sweden)

    Kei-ichi Ozaki

    2013-02-01

    Full Text Available It has generally been considered that protein phosphatases have more diverse catalytic domain structures and mechanisms than protein kinases; however, gene annotation efforts following the human genome project appeared to have completed the whole array of protein phosphatases. Ser/Thr phosphatases are divided into three subfamilies that have different structures from each other, whereas Tyr phosphatases and dual-specificity phosphatases targeting Tyr, Ser and Thr belong to a single large family based on their common structural features. Several years of research have revealed, however, the existence of unexpected proteins, designated here as “atypical protein phosphatases”, that have structural and enzymatic features different from those of the known protein phosphatases and are involved in important biological processes. In this review, we focus on the identification and functional characterization of atypical protein phosphatases, represented by eyes absent (EYA, suppressor of T-cell receptor signaling (Sts and phosphoglycerate mutase family member 5 (PGAM5 and discuss their biological significance in cellular signaling.

  11. Development of alkaline fuel cells.

    Energy Technology Data Exchange (ETDEWEB)

    Hibbs, Michael R.; Jenkins, Janelle E.; Alam, Todd Michael; Janarthanan, Rajeswari [Colorado School of Mines, Golden, CO; Horan, James L. [Colorado School of Mines, Golden, CO; Caire, Benjamin R. [Colorado School of Mines, Golden, CO; Ziegler, Zachary C. [Colorado School of Mines, Golden, CO; Herring, Andrew M. [Colorado School of Mines, Golden, CO; Yang, Yuan [Colorado School of Mines, Golden, CO; Zuo, Xiaobing [Argonne National Laboratory, Argonne, IL; Robson, Michael H. [University of New Mexico, Albuquerque, NM; Artyushkova, Kateryna [University of New Mexico, Albuquerque, NM; Patterson, Wendy [University of New Mexico, Albuquerque, NM; Atanassov, Plamen Borissov [University of New Mexico, Albuquerque, NM

    2013-09-01

    This project focuses on the development and demonstration of anion exchange membrane (AEM) fuel cells for portable power applications. Novel polymeric anion exchange membranes and ionomers with high chemical stabilities were prepared characterized by researchers at Sandia National Laboratories. Durable, non-precious metal catalysts were prepared by Dr. Plamen Atanassov's research group at the University of New Mexico by utilizing an aerosol-based process to prepare templated nano-structures. Dr. Andy Herring's group at the Colorado School of Mines combined all of these materials to fabricate and test membrane electrode assemblies for single cell testing in a methanol-fueled alkaline system. The highest power density achieved in this study was 54 mW/cm2 which was 90% of the project target and the highest reported power density for a direct methanol alkaline fuel cell.

  12. Myosin phosphatase is inactivated by caspase-3 cleavage and phosphorylation of myosin phosphatase targeting subunit 1 during apoptosis

    OpenAIRE

    Iwasaki, Takahiro; Katayama, Takeshi; Kohama, Kazuhiro; Endo, Yaeta; Sawasaki, Tatsuya

    2013-01-01

    This study indicates that myosin phosphatase in apoptotic cells is inactivated by the cleavage and the phosphorylation of myosin phosphatase targeting subunit 1 (MYPT1). The results suggest that down-regulation of MYPT1 promotes hyperphosphorylation of myosin II during apoptosis.

  13. Interaction of alkali and alkaline earth ions with Ochratoxin A

    Energy Technology Data Exchange (ETDEWEB)

    Poor, Miklos [Institute of Laboratory Medicine, University of Pecs, Pecs H-7624 (Hungary); Kunsagi-Mate, Sandor; Matisz, Gergely; Li, Yin; Czibulya, Zsuzsanna [Department of General and Physical Chemistry, University of Pecs, Pecs H-7624 (Hungary); Janos Szentagothai Research Center, Pecs H-7624 (Hungary); Peles-Lemli, Beata [Department of General and Physical Chemistry, University of Pecs, Pecs H-7624 (Hungary); Koszegi, Tamas, E-mail: koszegit@freemail.hu [Institute of Laboratory Medicine, University of Pecs, Pecs H-7624 (Hungary)

    2013-03-15

    The effect of alkali and alkaline earth ions on the chemical equilibrium of mono- and dianionic forms of the mycotoxin Ochratoxin A (OTA) and their bonding onto the surface of Bovine Serum Albumin (BSA) have been investigated by fluorescence spectroscopy and fluorescence polarization techniques. Our results show that alkali metal ions shift the chemical equilibrium towards formation of dianionic form of OTA. Furthermore, the alkaline earth ions can compete with BSA for binding to OTA when these ions are present in millimolar concentrations. Our data also highlight the possibility that the 'free' fraction of OTA (not bound onto the surface of albumin) or at least a part of it is present in cation-bound form in body fluids. These observations are supported by stability constants and quantum-chemical calculations. Among the studied alkaline metal ions magnesium showed the highest affinity towards OTA under physiological conditions. Further research is required to analyze the potential significance of Mg{sup 2+}-OTA complex in cellular uptake and/or elimination of the toxin in the human body. - Highlights: Black-Right-Pointing-Pointer Fluorescence spectroscopy reveals cation-Ochratoxin A (OTA) interactions. Black-Right-Pointing-Pointer Alkali ions shift the equilibrium of OTA to formation of a dianionic structure. Black-Right-Pointing-Pointer Alkaline earth ions directly bind to OTA in the order: Mg{sup 2+}, Ca{sup 2+}, Ba{sup 2+}. Black-Right-Pointing-Pointer Quantum chemical calculations and logK values support our experimental data.

  14. Inositol monophosphate phosphatase genes of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Parish Tanya

    2010-02-01

    Full Text Available Abstract Background Mycobacteria use inositol in phosphatidylinositol, for anchoring lipoarabinomannan (LAM, lipomannan (LM and phosphatidylinosotol mannosides (PIMs in the cell envelope, and for the production of mycothiol, which maintains the redox balance of the cell. Inositol is synthesized by conversion of glucose-6-phosphate to inositol-1-phosphate, followed by dephosphorylation by inositol monophosphate phosphatases (IMPases to form myo-inositol. To gain insight into how Mycobacterium tuberculosis synthesises inositol we carried out genetic analysis of the four IMPase homologues that are present in the Mycobacterium tuberculosis genome. Results Mutants lacking either impA (Rv1604 or suhB (Rv2701c were isolated in the absence of exogenous inositol, and no differences in levels of PIMs, LM, LAM or mycothiol were observed. Mutagenesis of cysQ (Rv2131c was initially unsuccessful, but was possible when a porin-like gene of Mycobacterium smegmatis was expressed, and also by gene switching in the merodiploid strain. In contrast, we could only obtain mutations in impC (Rv3137 when a second functional copy was provided in trans, even when exogenous inositol was provided. Experiments to obtain a mutant in the presence of a second copy of impC containing an active-site mutation, in the presence of porin-like gene of M. smegmatis, or in the absence of inositol 1-phosphate synthase activity, were also unsuccessful. We showed that all four genes are expressed, although at different levels, and levels of inositol phosphatase activity did not fall significantly in any of the mutants obtained. Conclusions We have shown that neither impA, suhB nor cysQ is solely responsible for inositol synthesis. In contrast, we show that impC is essential for mycobacterial growth under the conditions we used, and suggest it may be required in the early stages of mycothiol synthesis.

  15. Increased tartrate-resistant acid phosphatase (TRAP) expression in malignant breast, ovarian and melanoma tissue: an investigational study

    International Nuclear Information System (INIS)

    Tartrate-resistant acid phosphatase (TRAP) is a metalloprotein enzyme that belongs to the acid phosphatases and is known to be expressed by osteoclasts. It has already been investigated as a marker of bone metastases in cancer patients. In this study, which examined the value of serum TRAP concentrations as a marker of bone disease in breast cancer patients, we observed high concentrations of TRAP even in patients without bone metastases. To elucidate this phenomenon, we examined the expression of TRAP in breast cancer cells and the cells of several other malignancies. TRAP concentrations in the serum of tumor patients were determined by ELISA. The expression of TRAP in breast, ovarian, and cervical cancer and malignant melanoma was analyzed by immunohistochemistry. RT-PCR and immunocytology were used to evaluate TRAP expression in cultured tumor cells. A marked increase in serum TRAP concentrations was observed in patients with breast and ovarian cancer, regardless of the presence or absence of bone disease. TRAP expression was found in breast and ovarian cancers and malignant melanoma, while cervical cancer showed only minimal expression of TRAP. Expression of TRAP was absent in benign tissue or was much less marked than in the corresponding malignant tissue. TRAP expression was also demonstrated in cultured primary cancer cells and in commercially available cell lines. Overexpression of TRAP was detected in the cells of various different tumors. TRAP might be useful as a marker of progression of malignant disease. It could also be a potential target for future cancer therapies

  16. Protein electrophoresis - serum

    Science.gov (United States)

    This lab test measures the types of protein in the fluid (serum) part of a blood sample. Other electrophoresis tests that measure proteins in the serum include: Immunoelectrophoresis Immunofixation Globulin electrophoresis

  17. Zinc ions and alkaline pH alter the phosphorylation state of human erythrocyte membrane proteins

    International Nuclear Information System (INIS)

    Since the phosphorylation state of the red cell membrane proteins in vitro is likely to be regulated by phosphorylation and dephosphorylation, this research was carried out to investigate the possible role of membrane-bound phosphatase activities. These studies were conducted with red blood cell ghosts and IOVs from normal individuals and from an individual with hereditary spherocytosis. In vitro phosphorylation with (?-32P) ATP was conducted in the presence and the absence of Zn++, or erythrocyte ghosts and IOVs were pretreated for 30 minutes at 37 degree C and pH 7-11 in the presence and the absence of calf intestine alkaline phosphatase. The resulting phosphoproteins were analyzed by SDS-polyacrylamide gel electrophoresis, stained with Coomassie blue, and fluorographed. In the presence of Zn++, the red blood ghosts, with or without pretreatment, demonstrated enhanced phosphorylation of membrane proteins, including band 4.2. Preincubation at pH 10 in the presence of absence of exogenous phosphatase further stimulates phosphorylation of these proteins. Under similar conditions, the erythrocyte membranes also demonstrated the ability to hydrolyze p-nitrophenyl phosphate and to remove 32P from red blood cell phosphoproteins

  18. Dose-response relationship between arsenic exposure and the serum enzymes for liver function tests in the individuals exposed to arsenic: a cross sectional study in Bangladesh

    Directory of Open Access Journals (Sweden)

    Hossain Mostaque

    2011-07-01

    Full Text Available Abstract Background Chronic arsenic exposure has been shown to cause liver damage. However, serum hepatic enzyme activity as recognized on liver function tests (LFTs showing a dose-response relationship with arsenic exposure has not yet been clearly documented. The aim of our study was to investigate the dose-response relationship between arsenic exposure and major serum enzyme marker activity associated with LFTs in the population living in arsenic-endemic areas in Bangladesh. Methods A total of 200 residents living in arsenic-endemic areas in Bangladesh were selected as study subjects. Arsenic concentrations in the drinking water, hair and nails were measured by Inductively Coupled Plasma Mass Spectroscopy (ICP-MS. The study subjects were stratified into quartile groups as follows, based on concentrations of arsenic in the drinking water, as well as in subjects' hair and nails: lowest, low, medium and high. The serum hepatic enzyme activities of alkaline phosphatase (ALP, aspartate transaminase (AST and alanine transaminase (ALT were then assayed. Results Arsenic concentrations in the subjects' hair and nails were positively correlated with arsenic levels in the drinking water. As regards the exposure-response relationship with arsenic in the drinking water, the respective activities of ALP, AST and ALT were found to be significantly increased in the high-exposure groups compared to the lowest-exposure groups before and after adjustments were made for different covariates. With internal exposure markers (arsenic in hair and nails, the ALP, AST and ALT activity profiles assumed a similar shape of dose-response relationship, with very few differences seen in the higher groups compared to the lowest group, most likely due to the temporalities of exposure metrics. Conclusions The present study demonstrated that arsenic concentrations in the drinking water were strongly correlated with arsenic concentrations in the subjects' hair and nails. Further, this study revealed a novel exposure- and dose- response relationship between arsenic exposure metrics and serum hepatic enzyme activity. Elevated serum hepatic enzyme activities in the higher exposure gradients provided new insights into arsenic-induced liver toxicity that might be helpful for the early prognosis of arsenic-induced liver diseases.

  19. Marcadores del remodelamiento óseo en saliva y su correlación con los niveles sanguíneos en ratas Bone remodeling markers in saliva as compared to serum in rats

    Directory of Open Access Journals (Sweden)

    Pellegrini Gretel

    2006-06-01

    Full Text Available Si bien es conocida la utilidad de marcadores óseos en suero u orina para determinar cambios en el remodelamiento óseo, la misma no ha sido totalmente estudiada en saliva. Este trabajo evalúa la correlación entre dos marcadores del recambio óseo: la fosfatasa alcalina ósea (isoforma ósea, FAO y el telopéptido C-terminal del colágeno tipo I (CTX, medidos simultáneamente en suero y saliva de ratas Wistar (250 a 300 g, SHAM (n=12 y ovariectomizadas (OVX (n=12. Luego de una semana de la cirugía se extrajo sangre en ayunas y saliva total estimulada donde se evaluó CTX (ELISA, RatLabs, Osteometer Bio Tech, Dinamarca y FAO (Wiener, colorimetría. En el suero, tanto CTX (ng/ml como FAO (UI/l en ratas OVX fueron significativamente mayores que en ratas SHAM (15.3±4.0 vs. 21.8±6.4, pBone markers are useful tools to measure bone remodeling; currently they are assessed in serum and urinary samples; however there is little information concerning their measurement in saliva. The present experimental study evaluates the possibility to measure collagen type I carboxiterminal telopeptide (CTX and bone alkaline phosphatase (b-AP in saliva, its correlation with serum samples in normal conditions and in the increase of the bone remodeling due to estrogen deficiency. Twenty four normal adult Wistar rats (300±20 g [12 SHAM and 12 rats after 1 week of bilateral ovariectomy (OVX] were studied. Fasting serum and total saliva after stimulation with pilocarpine were collected. In both samples were measured: CTX (ng/ml by ELISA (RatLabs, Osteometer Bio Tech, Denmark and b-AP (IU/L (Wiener, colorimetrically. Both CTX and b-AL in serum samples were significantly higher in OVX than in SHAM rats (15.3±4.0 vs. 21.8±6.4, p<0.05 y 71±29 vs. 104±23; p<0.01, respectively. Saliva presented the same behaviour (3.6±0.5 vs. 6.4±2.9; p<0.02 y 73±29 vs. 90±8; p<0.003, respectively. When saliva CTX and b-AP were plotted against serum concentration significant positive correlations were obtained: r=0.58, p<0.05 and r=0.59; p<0.05, respectively. In conclusion, the present results are promisory in the sense of the potential use of a salivary-based test for evaluating bone remodeling. However, the use of this methodology for clinical practice needs extensive additional investigations.

  20. Association of Tartrate-Resistant Acid Phosphatase-Expressed Macrophages and Metastatic Breast Cancer Progression

    Science.gov (United States)

    Chen, Yu-Guang; Janckila, Anthony; Chao, Tsu-Yi; Yeh, Ren-Hua; Gao, Hong-Wei; Lee, Su-Huei; Yu, Jyh-Cherng; Liao, Guo-Shiou; Dai, Ming-Shen

    2015-01-01

    Abstract Infiltrating neutrophils, lymphocytes, macrophages, and cytokines constitute a state of chronic inflammation within the tumor microenvironment. Tartrate-resistant acid phosphatase 5a (TRACP5a) protein, a novel product of activated macrophage, is postulated to be a biomarker for systemic inflammatory burden in states of chronic inflammation. We aimed to investigate the clinical significance of TRACP5a expression in tumor-infiltrating macrophages and serum TRACP5a in patients with metastatic breast cancer (BC). We retrospectively analyzed the clinical data from 34 BC patients with confirmed skeletal/visceral metastasis upon or during first-line palliative treatment. Patients were stratified into 3 groups based on the therapeutic responses and follow-up disease course. The association of TRACP5a protein with other inflammatory and cancer biomarkers was assessed among the clinically distinct group of patients. Higher TRACP5a protein was significantly correlated with earlier disease progression and survival (P?=?0.0045) in comparison to other inflammatory markers, CRP or IL-6. Patients with higher serum TRACP5a level and shorter survival and treatment refractoriness also had more TRACP+ tumor-infiltrating macrophages. Our data support a hypothesis that serum TRACP5a protein can potentially be a predictive and prognostic marker to evaluate disease progression and therapeutic response in BC patients with bone/visceral metastasis. The associations between overall survival and TRACP expression by macrophages require further prospective investigation. PMID:26632898

  1. Moraxella catarrhalis Synthesizes an Autotransporter That Is an Acid Phosphatase?

    OpenAIRE

    Hoopman, Todd C.; Wang, Wei; Brautigam, Chad A; Sedillo, Jennifer L.; Reilly, Thomas J.; Hansen, Eric J.

    2007-01-01

    Moraxella catarrhalis O35E was shown to synthesize a 105-kDa protein that has similarity to both acid phosphatases and autotransporters. The N-terminal portion of the M. catarrhalis acid phosphatase A (MapA) was most similar (the BLAST probability score was 10?10) to bacterial class A nonspecific acid phosphatases. The central region of the MapA protein had similarity to passenger domains of other autotransporter proteins, whereas the C-terminal portion of MapA resembled the translocation dom...

  2. Alkaline fuel cell performance investigation

    International Nuclear Information System (INIS)

    An exploratory experimental fuel cell test program was conducted to investigate the performance characteristics of alkaline laboratory research electrodes. The objective of this work was to establish the effect of temperature, pressure, and concentration upon performance and evaluate candidate cathode configurations having the potential for improved performance. The performance characterization tests provided data to empirically establish the effect of temperature, pressure, and concentration upon performance for cell temperatures up to 300 F and reactant pressures up to 200 psia. Evaluation of five gold alloy cathode catalysts revealed that three doped gold alloys had more that two times the surface areas of reference cathodes and therefore offered the best potential for improved performance

  3. Alkaline fuel cell performance investigation

    Science.gov (United States)

    Martin, R. E.; Manzo, M. A.

    1988-01-01

    An exploratory experimental fuel cell test program was conducted to investigate the performance characteristics of alkaline laboratory research electrodes. The objective of this work was to establish the effect of temperature, pressure, and concentration upon performance and evaluate candidate cathode configurations having the potential for improved performance. The performance characterization tests provided data to empirically establish the effect of temperature, pressure, and concentration upon performance for cell temperatures up to 300 F and reactant pressures up to 200 psia. Evaluation of five gold alloy cathode catalysts revealed that three doped gold alloys had more that two times the surface areas of reference cathodes and therefore offered the best potential for improved performance.

  4. Valores bioquímicos séricos de tilapia del nilo (oreochromis niloticus) en cultivo intensivo / Serum biochemical values for nile tilapia (oreochromis niloticus) cultured under an intensive system

    Scientific Electronic Library Online (English)

    Leandro, Z. Crivelenti; Sofia, Borín; José Javier, M. Socha; Antonio V, Mundim.

    2011-12-01

    Full Text Available Objetivou-se a avaliação dos parâmetros bioquímicos séricos de tilápias do Nilo (Oreochromis niloticus), linhagem tailandesa Chitralada, produzidas em sistema de piscicultura intensiva e capturadas por tarrafeamento. Amostras sanguíneas de 40 exemplares, pesando em média 453 ± 52 g, foram obtidas po [...] r venopunção caudal e posteriormente analisadas quanto às concentrações séricas de proteína total, albumina, globulinas, ácido úrico, creatinina, ureia, cálcio, fósforo inorgânico, relação Ca/P, magnésio e fosfatase alcalina. Em um contexto geral, os resultados evidenciaram parâmetros semelhantes aos estabelecidos para peixes de escama, com exceção do ácido úrico. Abstract in spanish El objetivo del estudio fue evaluar los parámetros bioquímicos séricos de tilapias del Nilo (Oreochromis niloticus), linaje tailandés chitralada, producidas en un sistema de piscicultura intensiva y capturadas con atarraya. Se tomaron muestras de sangre a 40 ejemplares por punción de la vena caudal. [...] Los peces tenían un peso de 453 ± 52 g. Se determinó proteína total, albúmina, globulinas, ácido úrico, creatinina, urea, calcio, fósforo inorgánico, relación Ca/P, magnesio y fosfatasa alcalina. En un contexto general, los resultados mostraron parámetros semejantes a los establecidos para peces de escama, con excepción del ácido úrico. Abstract in english The aim of the study was to evaluate the serum biochemical parameters of the Nile tilapia (Oreochromis niloticus), Thai Chitralada lineage, raised under an intensive pisciculture system and captured by the cast net technique. Blood samples from 40 individuals were obtained by caudal venopuncture. Me [...] an body weight was 453 ± 52 g. The serum biochemical parameters evaluated were total protein, albumin, globulins, acid uric, creatinine, urea, calcium, inorganic phosphorus, Ca/P ratio, alkaline phosphatase, and magnesium. In general, the data showed similar parameters in comparison to those established for scale fish, exception of the acid uric.

  5. Impact of growth conditions on susceptibility of five microbial species to alkaline stress

    OpenAIRE

    Brändle, N; Zehnder, M; Weiger, R.; Waltimo, T.

    2008-01-01

    The effects of different growth conditions on the susceptibility of five taxa to alkaline stress were investigated. Enterococcus faecalis ATCC 29212, Streptococcus sobrinus OMZ 176, Candida albicans ATCC 90028, Actinomyces naeslundii ATCC 12104, and Fusobacterium nucleatum ATCC 10953 were grown as planktonic cells, allowed to adhere to dentin for 24 hours, grown as monospecies or multispecies biofilms on dentin under anaerobic conditions with a serum-enriched nutrient supply at 37 degrees C f...

  6. Changes of the activity of certain lysosomal enzymes in the blood serum of whole-body irradiated rats

    International Nuclear Information System (INIS)

    The activities of acid phosphatase, ?-glucuronidase, ?-galactosidase, acid ribonuclease, and acid deoxyribonuclease were studied in the blood serum of rats after total, either single or fractionated, exposure. After the single, total exposure to 800 R of X-rays, remarkable increases in the activities of acid phosphatase and acid deoxyribonuclease were observed in the blood serum immediately after the irradiation. At later stages were observed statistically significant decreases of ?-glucuronidase and ?-galactosidase in the rat blood serum after the total, single exposure. The serum acid ribonuclease activity remained essentially unaltered over the whole time interval of interest. In the blood serum of the rats exposed to total, fractionated irradiation, statistically significant decreases in the acid phosphatase and ?-glucuronidase activities were observed 1 and 8 days after completing the irradiation. In the case of ?--galactosidase, this decrease lasted even up to the 15th day after the end of irradiation. The activities of serum acid deoxyribonuclease and acid ribonuclease exhibited no statistically significant changes. (orig.)

  7. Acid phosphatase and lipid peroxidation in human cataractous lens epithelium

    OpenAIRE

    Vasavada Abhay; Thampi Prajitha; Yadav Savita; Rawal U

    1993-01-01

    The anterior lens epithelial cells undergo a variety of degenerative and proliferative changes during cataract formation. Acid phosphatase is primarily responsible for tissue regeneration and tissue repair. The lipid hydroperoxides that are obtained by lipid peroxidation of polysaturated or unsaturated fatty acids bring about deterioration of biological membranes at cellular and tissue levels. Acid phosphatase and lipid peroxidation activities were studied on the lens epithelial cells of nucl...

  8. Haptoglobin, acid phosphatase and demographic factors: obesity risk

    OpenAIRE

    Ramos, Maria do Rosário; Carolino, Elisabete; Oliveira, Teresa; Silva, A. P.; Carvalho, R.; Bicho, M.

    2009-01-01

    The aim of this work is to study the risk of obesity posed by two genetic factors: haptoglobin phenotype and acid phosphatase phenotype, one enzymatic activity: acid phosphatase activity (ACP1), age and gender. Haptoglobin (Hp) is a protein of the immune system, and three phenotypes of Hp are found in humans: Hp1-1, Hp2-1, and Hp2-2. This protein is associated with a susceptibility to common pathological conditions, such as obesity. ...

  9. Acid phosphatases in seeds and developing of squash (Cucurbita ficifolia)

    OpenAIRE

    Irena Lorenc-Kubis

    1994-01-01

    Changes in protein content and acid phosphatase activity were followed during germination (imbition through seedlings development) in extracts from cotyledons of squash (Cucurbita ficifolia). It has been shown that the activity of acid phosphatase was initially low and than increased to a maximum after 6 days of imbition. Acid phosphates were isolated from cotyledons of seeds and from 6-, 10- and 22-days old seedlings by extraction the proteins with 0.1 M acetate buffer pH 5.1, precipitation ...

  10. Acid phosphatase activity in Coxiella burnetii: a possible virulence factor.

    OpenAIRE

    Baca, O. G.; Roman, M.J.; Glew, R. H.; Christner, R F; Buhler, J E; Aragon, A S

    1993-01-01

    High-speed supernatant fluids derived from sonicated Coxiella burnetii contained considerable acid phosphatase activity when assayed by using 4-methylumbelliferylphosphate; they also contained a factor that blocked superoxide anion production by human neutrophils stimulated with formyl-Met-Leu-Phe. The pH optimum of the enzyme was approximately 5.0. The level of phosphatase activity detected in several isolates of C. burnetii implicated in acute (Nine Mile) and chronic (S Q217, PRS Q177, K Q1...

  11. The end of mitosis from a phosphatase perspective

    OpenAIRE

    Visconti, Roberta; Palazzo, Luca; Pepe, Anna; Monica, Rosa Della; Grieco, Domenico

    2013-01-01

    Transition through mitosis, the cell division cycle phase deputed to segregate replicated chromosomes, requires a wave of protein phosphorylation. While in the past decades a wealth of information has been gathered on the major kinase activities responsible for the onset of mitosis, only recently has a picture emerged of how their effects are reversed by protein phosphatases at the end of mitosis. Here, we summarized some recent data on the relevance for protein phosphatases in the reversal o...

  12. High-resolution colorimetric assay for rapid visual readout of phosphatase activity based on gold/silver core/shell nanorod.

    Science.gov (United States)

    Gao, Zhuangqiang; Deng, Kaichao; Wang, Xu-Dong; Miró, Manuel; Tang, Dianping

    2014-10-22

    Nanostructure-based visual assay has been developed for determination of enzymatic activity, but most involve in poor visible color resolution and are not suitable for routine utilization. Herein, we designed a high-resolution colorimetric protocol based on gold/silver core/shell nanorod for visual readout of alkaline phosphatase (ALP) activity by using bare-eyes. The method relied on enzymatic reaction-assisted silver deposition on gold nanorod to generate significant color change, which was strongly dependent on ALP activity. Upon target ALP introduction into the substrate, the ascorbic acid 2-phosphate was hydrolyzed to form ascorbic acid, and then, the generated ascorbic acid reduced silver ion to metal silver and coated on the gold nanorod, thereby resulting in the blue shift of longitudinal localized surface plasmon resonance peak of gold nanorod accompanying a perceptible color change from red to orange to yellow to green to cyan to blue and to violet. Under optimal conditions, the designed method exhibited the wide linear range 5-100 mU mL(-1) ALP with a detection limit of 3.3 mU mL(-1). Moreover, it could be used for the semiquantitative detection of ALP from 20 to 500 mU mL(-1) by using the bare-eyes. The coefficients of variation for intra- and interassay were below 3.5% and 6.2%, respectively. Finally, this method was validated for the analysis of real-life serum samples, giving results matched well with those from the 4-nitrophenyl phosphate disodium salt hexahydrate (pNPP)-based standard method. In addition, the system could even be utilized in the enzyme-linked immunosorbent assay (ELISA) to detect IgG at picomol concentration. With the merits of simplification, low cost, user-friendliness, and sensitive readout, the gold nanorod-based colorimetric assay has the potential to be utilized by the public and opens a new horizon for bioassays. PMID:25244147

  13. Ionized alkaline water: new strategy for management of metabolic acidosis in experimental animals.

    Science.gov (United States)

    Abol-Enein, Hassan; Gheith, Osama A; Barakat, Nashwa; Nour, Eman; Sharaf, Abd-Elhameed

    2009-06-01

    Metabolic acidosis can occur as a result of either the accumulation of endogenous acids or loss of bicarbonate from the gastrointestinal tract or the kidney, which represent common causes of metabolic acidosis. The appropriate treatment of acute metabolic acidosis has been very controversial. Ionized alkaline water was not evaluated in such groups of patients in spite of its safety and reported benefits. So, we aimed to assess its efficacy in the management of metabolic acidosis in animal models. Two models of metabolic acidosis were created in dogs and rats. The first model of renal failure was induced by ligation of both ureters; and the second model was induced by urinary diversion to gut (gastrointestinal bicarbonate loss model). Both models were subjected to ionized alkaline water (orally and by hemodialysis). Dogs with renal failure were assigned to two groups according to the type of dialysate utilized during hemodialysis sessions, the first was utilizing alkaline water and the second was utilizing conventional water. Another two groups of animals with urinary diversion were arranged to receive oral alkaline water and tap water. In renal failure animal models, acid-base parameters improved significantly after hemodialysis with ionized alkaline water compared with the conventional water treated with reverse osmosis (RO). Similar results were observed in urinary diversion models as there was significant improvement of both the partial pressure of carbon dioxide and serum bicarbonate (P = 0.007 and 0.001 respectively) after utilizing alkaline water orally. Alkaline ionized water can be considered as a major safe strategy in the management of metabolic acidosis secondary to renal failure or dialysis or urinary diversion. Human studies are indicated in the near future to confirm this issue in humans. PMID:19527469

  14. Activation analysis of alkaline rocks

    International Nuclear Information System (INIS)

    The United States Geological Survey reference sample AGV-1 andesite and three alkaline rocks from the apatite mine of Jacupiranga, Brasil, were analysed by thermal neutron activation analysis using destructive an non-destructive methods and high resolution Ge(Li) gamma-ray detectors. One of the rocks, a carbonite, was also analysed by instrumental activation analysis with epithermal neutrons. A greater number of elements can be determined using the radiochemical separation, but the precision and accuracy attained by INAA and RNAA were of the same order for most of the elements analysed. Epithermal activation was more advantageous for tantalum, terbium and holmium. Comparison of the analytical results for USGS reference sample (AGV-1) with the data published by others gave good agreement. Sta tistical tests used for comparison of the results of destructive and non-destructive methods, are presented. Gamma spectra are shown and tabulated data are given for more than 25 elements. (T.G.)

  15. Alkaline azide mutagenicity in cowpea

    International Nuclear Information System (INIS)

    Sodium azide is known as a potent mutagen in cereals and legumes. It is very effective in acidic medium in barley. Here an attempt is made to measure the effectiveness of sodium azide in alkaline medium (pH 7.4) on cowpea (Vigna unguiculata (L.) Walp., variety FS-68). Seeds pre-soaked in distilled water for 5 hours were treated with different concentrations (10-6, 10-5, 10-4 and 10-3M) of sodium azide (NaN3) for 4 hours at 28± 2 deg. C. Bottles were intermittently shaken, then the seeds were thoroughly washed in running tap water and subsequently planted in pots. The treatment caused significant biological damage such as reduction in seed germination, length of root and shoot, number of nodules and pods per plant and morphological leaf variations. Morphological, as well as chlorophyll mutants, were detected in M2

  16. Serum oxidative stress is increased in patients with post cholecystectomy bile duct injury / Aumento del estrés oxidativo en el suero de pacientes con lesiones de vías biliares postcolecistectomía

    Scientific Electronic Library Online (English)

    A. G., Miranda-Díaz; J. M., Hermosillo-Sandoval; G. G., Ortiz; D., Lizardi-García; E. G., Cardona-Muñoz; F., Pacheco-Moisés.

    2010-06-01

    Full Text Available Introducción: las lesiones de las vías biliares postcolecistectomía se establecen por la aparición de ictericia, elevación de las bilirrubinas y de la fosfatasa alcalina durante el periodo perioperatorio. Se desconoce cómo se modifican los marcadores de estrés oxidativo en el suero de los pacientes [...] con lesiones de las vías biliares postcolecistectomía. Objetivo: determinar los marcadores de estrés oxidativo (productos de peroxidación de lípidos, catabolitos del óxido nítrico y capacidad antioxidante total) en el suero de pacientes con lesiones de las vías biliares. Pacientes y métodos: se realizó un estudio prospectivo transversal analítico. Se formaron 2 grupos de estudio. Grupo 1: 5 voluntarios sanos que sirvieron como control. Grupo 2: 52 pacientes con lesiones de las vías biliares postcolecistectomía (43 femeninos y 9 masculinos). La reconstrucción bilio-digestiva se realizó de manera electiva a las 8 semanas. Los niveles séricos de los marcadores de estrés oxidativo se cuantificaron mediante métodos colorimétricos. Resultados: los niveles séricos de los productos de peroxidación de lípidos (malondialdehído y 4-hidroxialquenos) y de los catabolitos del óxido nítrico (nitritos/nitratos) aumentaron significativamente en los pacientes con lesiones de las vías biliares en comparación a los voluntarios sanos. En contraste, la capacidad antioxidante total en el suero de los pacientes con lesiones de las vías biliares fue similar a la de los voluntarios sanos. Conclusiones: los resultados muestran que el estrés oxidativo en suero es un componente que se asocia a las lesiones de las vías biliares. Abstract in english Background: post-cholecystectomy bile duct injuries are identified by the onset of jaundice as well as elevated bilirubin and alkaline phosphatase levels during the peri-operative period. It is unknown how serum oxidative stress markers are modified in patients with post-cholecystectomy bile duct in [...] juries. Objective: to determine serum oxidative stress marker levels (lipid peroxidation by-products, nitrites/nitrates and total antioxidant capacity) in patients with post-cholecystectomy bile duct injuries. Patients and methods: a prospective, transversal and analytical study was designed with two groups. Group 1: 5 healthy volunteer subjects. Group 2: 52 patients with post-cholecystectomy bile duct injuries (43 female and 9 male). An elective bilio-digestive reconstruction was performed at week 8. The serum oxidative stress marker levels were quantified by colorimetric method. Results: patients with bile duct injuries had a significant increased serum lipid peroxides (malondialdehyde and 4-hydroxy-alkenals) and nitric oxide metabolites (nitrites/nitrates) levels compared to the control group. In contrast, total antioxidant capacity in patients with bile duct injuries remained similar compared to healthy controls. Conclusions: the results show that oxidative stress is usually associated to bile duct injury.

  17. Replacement of inorganic zinc with lower levels of organic zinc (zinc nicotinate on performance, hematological and serum biochemical constituents, antioxidants status, and immune responses in rats

    Directory of Open Access Journals (Sweden)

    D. Nagalakshmi

    2015-09-01

    Full Text Available Aim: A study was undertaken to investigate the effect of organic zinc (zinc nicotinate, Zn-nic supplementation (6, 9, and 12 ppm compared to inorganic zinc (12 ppm on growth performance, hematology, serum biochemical constituents oxidative stress, and immunity in weaned female Sprague–Dawley rats. Material and Methods: A 48 weaned rats (285.20±1.95 g were randomly distributed to 4 dietary treatments with 6 replicates in each and reared in polypropylene cages for 10 weeks. Basal diet (BD was formulated with purified ingredients without zinc (Zn. Four dietary treatments were prepared by adding 12 ppm Zn from ZnCO3 (control and 6, 9, and 12 ppm Zn from Zn-nic to the BD. On 42nd day, blood was collected by retro-orbital puncture for analyzing hematological constituents, glucose, cholesterol, alkaline phosphatase, total protein, albumin, and globulin and antioxidant enzyme activities. At 43rd day, rats were antigenically challenged with sheep red blood cell (RBC to assess humoral immune response and on 70th day cell-mediated immune response. Results: Weekly body weight gains, daily feed intake, blood hematological constituents (white blood cell, RBC, hemoglobin concentration, packed cell volume, mean corpuscular volume, lymphocyte, monocyte, and granulocyte concentration and serum glucose, total protein levels were comparable among the rats feed Zn from ZnCO3 and Zn-nic (6, 9, and 12 ppm. Serum cholesterol reduced with organic Zn supplementation at either concentration (6-12 ppm. Serum globulin concentration reduced (p<0.05 with 6 ppm Zn-nic supplementation compared to other dietary treatments. Lipid peroxidation lowered (p<0.05 reduced with 12 ppm organic Zn; thiobarbituric acid reacting substances and protein carbonyls concentrations in liver reduced (p<0.05 with 9 and 12 ppm levels of organic Zn supplementation compared to 12 ppm Zn supplementation from inorganic source. RBC catalase and glutathione peroxidase enzymes activities were highest (p<0.05 in rats supplemented with 12 ppm Zn-nic, followed by 9 ppm. Comparable immune response (humoral and cell-mediated was observed between 12 ppm inorganic Zn and 9 ppm organic Zn and higher (p<0.05 immune response was noticed at 12 ppm Zn-nic supplementation. Conclusion: Based on the results, it is concluded that dietary Zn concentration can be reduced by 50% (6 ppm as Zn nicotinate without affecting growth performance, hemato-biochemical constituents, antioxidant status, and immunity. In addition, replacement of 12 ppm inorganic Zn with 12 ppm organic Zn significantly improved antioxidant status and immune response.

  18. Maternal serum screening.

    OpenAIRE

    Klein, M. C.

    1994-01-01

    Maternal serum screening (MSS) measures three serum markers: alpha-fetoprotein, human chorionic gonadotropin, and unconjugated estriol, from which the risk of fetal Down syndrome or open neural tube defect is calculated. Initially, 8% of women will have positive results. I present a protocol for investigating these women. Family physicians should be informed about MSS so they can give their patients information and guidance.

  19. Acid phosphatase and lipid peroxidation in human cataractous lens epithelium

    Directory of Open Access Journals (Sweden)

    Vasavada Abhay

    1993-01-01

    Full Text Available The anterior lens epithelial cells undergo a variety of degenerative and proliferative changes during cataract formation. Acid phosphatase is primarily responsible for tissue regeneration and tissue repair. The lipid hydroperoxides that are obtained by lipid peroxidation of polysaturated or unsaturated fatty acids bring about deterioration of biological membranes at cellular and tissue levels. Acid phosphatase and lipid peroxidation activities were studied on the lens epithelial cells of nuclear cataract, posterior subcapsular cataract, mature cataract, and mixed cataract. Of these, mature cataractous lens epithelium showed maximum activity for acid phosphatase (516.83 moles of p-nitrophenol released/g lens epithelium and maximum levels of lipid peroxidation (86.29 O.D./min/g lens epithelium. In contrast, mixed cataractous lens epithelium showed minimum activity of acid phosphatase (222.61 moles of p-nitrophenol released/g lens epithelium and minimum levels of lipid peroxidation (54.23 O.D./min/g lens epithelium. From our study, we correlated the maximum activity of acid phosphatase in mature cataractous lens epithelium with the increased areas of superimposed cells associated with the formation of mature cataract. Likewise, the maximum levels of lipid peroxidation in mature cataractous lens epithelium was correlated with increased permeability of the plasma membrane. Conversely, the minimum levels of lipid peroxidation in mixed cataractous lens epithelium makes us presume that factors other than lipid peroxidation may also account for the formation of mixed type of cataract.

  20. Protein phosphatase 2A regulatory subunit B56? limits phosphatase activity in the heart.

    Science.gov (United States)

    Little, Sean C; Curran, Jerry; Makara, Michael A; Kline, Crystal F; Ho, Hsiang-Ting; Xu, Zhaobin; Wu, Xiangqiong; Polina, Iuliia; Musa, Hassan; Meadows, Allison M; Carnes, Cynthia A; Biesiadecki, Brandon J; Davis, Jonathan P; Weisleder, Noah; Györke, Sandor; Wehrens, Xander H; Hund, Thomas J; Mohler, Peter J

    2015-07-21

    Protein phosphatase 2A (PP2A) is a serine/threonine-selective holoenzyme composed of a catalytic, scaffolding, and regulatory subunit. In the heart, PP2A activity is requisite for cardiac excitation-contraction coupling and central in adrenergic signaling. We found that mice deficient in the PP2A regulatory subunit B56? (1 of 13 regulatory subunits) had altered PP2A signaling in the heart that was associated with changes in cardiac physiology, suggesting that the B56? regulatory subunit had an autoinhibitory role that suppressed excess PP2A activity. The increase in PP2A activity in the mice with reduced B56? expression resulted in slower heart rates and increased heart rate variability, conduction defects, and increased sensitivity of heart rate to parasympathetic agonists. Increased PP2A activity in B56?(+/-) myocytes resulted in reduced Ca(2+) waves and sparks, which was associated with decreased phosphorylation (and thus decreased activation) of the ryanodine receptor RyR2, an ion channel on intracellular membranes that is involved in Ca(2+) regulation in cardiomyocytes. In line with an autoinhibitory role for B56?, in vivo expression of B56? in the absence of altered abundance of other PP2A subunits decreased basal phosphatase activity. Consequently, in vivo expression of B56? suppressed parasympathetic regulation of heart rate and increased RyR2 phosphorylation in cardiomyocytes. These data show that an integral component of the PP2A holoenzyme has an important inhibitory role in controlling PP2A enzyme activity in the heart. PMID:26198358

  1. Alkaline rocks and the occurrence of uranium

    International Nuclear Information System (INIS)

    Many alkaline complexes contain uranium and other minerals in low concentrations and are regarded as constituting valuable potential reserves. Certain complex metallurgical problems, however, remain to be solved. Alkaline rocks occur in a number of forms and environments and it is noted that they are generated during periods of geological quiescence emplaced mainly in stable aseismic areas. Many occur along the extensions of oceanic transform faults beneath the continental crust and the application of this concept to areas not currently known to host alkaline complexes may prove useful in identifying potential target areas for prospecting operations

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

    OpenAIRE

    Peifang Wang; Lingxiao Ren; Chao(Saul) Wang; Jin Qian; Jun Hou

    2014-01-01

    Phosphorus (P) is an important limiting nutrient in aquatic ecosystems and knowledge of P cycling is fundamental for reducing harmful algae blooms and other negative effects in water. Despite their importance, the characteristics of P cycling under changing nutrient conditions in shallow lakes were poorly investigated. In this study, in situ incubation experiments were conducted in a natural riparian zone in the main diversion channel used for water transfer into Lake Taihu (Wangyu River). Va...

  3. LOW DIETARY COPPER INCREASES FECAL FREE RADICAL PRODUCTION AND FECAL WATER ALKALINE PHOSPHATASE ACTIVITY AND CYTOTOXICITY IN HEALTHY MEN

    Science.gov (United States)

    Background: One possible dietary factor that may increase susceptibility to colon cancer is inadequate copper intake. Objective: To investigate the effects of low and adequate copper intakes on copper nutriture and putative risk factors for colon cancer susceptibility in healthy men. Design: Sevente...

  4. Alkaline phosphatase activity at the southwest coast of India: A comparison of locations differently affected by upwelling..

    Digital Repository Service at National Institute of Oceanography (India)

    Mamatha, S.S.; Malik, A.; Varik, S.; Parvathi, V.; Jineesh, V.K.; Gauns, M.; LokaBharathi, P.A.

    The realization of the potential importance of phosphorus (P) as a limiting nutrient in marine ecosystem is increasing globally. Hence, the contribution of biotic variables in mobilizing this nutrient would be relevant especially in productive...

  5. Influence of Protein Conformation and Adjuvant Aggregation on the Effectiveness of Aluminum Hydroxide Adjuvant in a Model Alkaline Phosphatase Vaccine

    OpenAIRE

    Clausi, Amber L.; Morin, Andrea; CARPENTER, JOHN F.; Randolph, Theodore W

    2009-01-01

    The mechanism(s) of the enhancement of the immune response by addition of aluminum salt adjuvants to parenterally administered protein-based vaccines is still the subject of debate. It has been hypothesized, however, that destabilization of the antigen structure on the surface of the adjuvant may be important for eliciting immune response. Also, it has been suggested that immune response to adjuvanted vaccines is reduced if the adjuvant particles become aggregated before administration becaus...

  6. Host Plant Effects on Alkaline Phosphatase Activity in the Whiteflies, Bemisia tabaci Biotype B and Trialeurodes vaporariorum

    OpenAIRE

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

    2011-01-01

    Bemisia tabaci (Gennadius) B-biotype and Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae) often coexist on greenhouse-grown vegetable crops in northern China. The recent spread of B. tabaci B-biotype has largely replaced T. vaporariorum, and B-biotype now overlaps with T. vaporariorum where common hosts occur in most invaded areas. The impact of the B-biotype on the agro eco system appears to be widespread, and involves the ability to compete with and perhaps replace other phytop...

  7. Yam (Dioscorea batatas) Root and Bark Extracts Stimulate Osteoblast Mineralization by Increasing Ca and P Accumulation and Alkaline Phosphatase Activity

    OpenAIRE

    Kim, Suji; Shin, Mee-Young; Son, Kun-Ho; Sohn, Ho-Yong; Lim, Jae-Hwan; Lee, Jong-Hwa; Kwun, In-Sook

    2014-01-01

    Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within 0~10 ?g/mL during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineral...

  8. Association of erythrocyte acid phosphatase phenotypes with myopia

    Directory of Open Access Journals (Sweden)

    Himabindu P

    2005-01-01

    Full Text Available Acid phosphatase is a polymorphic nonspecific orthophosphate monoesterase which catalyses the cleaving of phosphoric acid and subsequent breakdown of several monophosphoric esters under acidic pH conditions. Acid phosphatase has a physiologic function as a flavin mononucleotide phosphatase (FMN and regulates the intracellular concentrations of flavin coenzymes that are electron carriers in the oxidative phosphorylation pathway. Myopia or nearsightedness is caused by both environmental and genetic factors. Myopic eyes when subjected to excessive oxidative stress results in retinal detachments .In the present study there is a significant elevation of AA phenotype in myopes when compared to controls. The AA phenotype is more susceptible to oxidative stress and its lower enzyme activity is known to be associated with increased intrauterine growth that further results in increased axial length in progressive myopia. The AA phenotype also confers risk for myopia development in males, early age group and cases with parental consanguinity.

  9. Alkalinity in oil field waters - what alkalinity is and how it is measured

    International Nuclear Information System (INIS)

    The alkalinity is an important parameter in the description of pH-behaviour, buffer capacity and scaling potentials in oil field waters. Although the alkalinity is widely used, it seems to be considerable confusion in connection with the concept. It is often used incorrectly and different authors define the concept in different ways. Several different methods for the determination of alkalinity can be found in the literature. This paper discusses the definition of alkalinity and how to use alkalinity in oil field waters to obtain data of importance for scale and pH predictions. There is also shown how a simple titration of oil field waters can give both the alkalinity and the content of organic acids in these waters. It is obvious from these findings that most of the methods used to day may give considerable errors when applied to oil field waters with high contents of organic acids. 8 refs., 8 figs., 5 tabs

  10. Structural Basis of Response Regulator Dephosphorylation by Rap Phosphatases

    Energy Technology Data Exchange (ETDEWEB)

    V Parashar; N Mirouze; D Dubnau; M Neiditch

    2011-12-31

    Bacterial Rap family proteins have been most extensively studied in Bacillus subtilis, where they regulate activities including sporulation, genetic competence, antibiotic expression, and the movement of the ICEBs1 transposon. One subset of Rap proteins consists of phosphatases that control B. subtilis and B. anthracis sporulation by dephosphorylating the response regulator Spo0F. The mechanistic basis of Rap phosphatase activity was unknown. Here we present the RapH-Spo0F X-ray crystal structure, which shows that Rap proteins consist of a 3-helix bundle and a tetratricopeptide repeat domain. Extensive biochemical and genetic functional studies reveal the importance of the observed RapH-Spo0F interactions, including the catalytic role of a glutamine in the RapH 3-helix bundle that inserts into the Spo0F active site. We show that in addition to dephosphorylating Spo0F, RapH can antagonize sporulation by sterically blocking phosphoryl transfer to and from Spo0F. Our structure-function analysis of the RapH-Spo0F interaction identified Rap protein residues critical for Spo0F phosphatase activity. This information enabled us to assign Spo0F phosphatase activity to a Rap protein based on sequence alone, which was not previously possible. Finally, as the ultimate test of our newfound understanding of the structural requirements for Rap phosphatase function, a non-phosphatase Rap protein that inhibits the binding of the response regulator ComA to DNA was rationally engineered to dephosphorylate Spo0F. In addition to revealing the mechanistic basis of response regulator dephosphorylation by Rap proteins, our studies support the previously proposed T-loop-Y allostery model of receiver domain regulation that restricts the aromatic 'switch' residue to an internal position when the {beta}4-{alpha}4 loop adopts an active-site proximal conformation.

  11. Perfil bioquímico e nutricional do ácido glutâmico e da vitamina K no soro e no fígado de frangos de corte de 1 a 21 dias de idade / Biochemical and nutritional profile of glutamic acid and vitamin K in serum and liver of broilers from 1 to 21 days

    Scientific Electronic Library Online (English)

    Cibele Silva, Minafra; George Henrique Kling de, Moraes; Ana Cláudia Peres, Rodrigues; Fernanda Alvares da, Silva; José Henrique, Stringhini; Cíntia Silva Minafra e, Rezende.

    2008-11-01

    Full Text Available Foram investigados os efeitos nutricionais de dois níveis de ácido L-glutâmico (L-Glu) combinados a quatro níveis de vitamina K (vit. K) no soro e no fígado de pintos de corte machos, Hubbard, de 1 dia de idade, criados em baterias aquecidas recebendo dieta básica purificada suplementada com 6,25 e [...] 12,5% de ácido glutâmico combinados a 0,02; 0,2; 2,0 e 20,0 mg de vit. K/kg. Duzentas e cinqüenta e seis aves foram distribuídas em esquema fatorial 2 × 4, em delineamento inteiramente casualizado, com quatro repetições de oito aves. As concentrações de cálcio e a fosfatase alcalina (FA) no soro não diferiram aos 7, 14 e 21 dias de idade. Os níveis séricos de fósforo diferiram aos 21 dias de idade e foram maiores com 12,5% de ácido glutâmico. O peso de fígado foi maior nas aves alimentadas com a dieta com 6,25% de ácido glutâmico e 12,5% de ácido glutâmico + 0,02 mg vit. K/kg. Os níveis séricos de proteínas totais mantiveram-se muito abaixo do normal. As concentrações da enzima glutamato-piruvato transaminase (GPT) foram elevadas nas aves alimentadas com ácido glutâmico e vit. K nos níveis avaliados. A enzima alanina aminotransferase foi encontrada em concentração muito superior aos níveis considerados normais para aves alimentadas com dietas contendo ácido glutâmico e vitamina K nos níveis avaliados. Os níveis de vit. K estudados não afetaram a absorção de cálcio e fósforo e da enzima fosfatase alcalina nem o metabolismo do tecido ósseo. O nível de nitrogênio não-específico (ácido glutâmico) de 6,25% das dietas alterou o metabolismo do tecido ósseo, como resultado da maior concentração de fosfatase alcalina no soro das aves. Abstract in english Nutritional effects of two levels of L-glutamic acid (L-Glu) combined with four levels of vitamin K (vit. K) in serum and the liver of chicks of day-old broilers, male, Hubbard, raised in brooded batteries, receiving basic purified diet supplemented with 6.25 and 12.5% of L-Glu combined with 0.02, 0 [...] .2, 2.0 and 20.0 mg vit. K/kg. Two hundred and fifty birds were allotted to completely randomized design an in 2 × 4 factorial arrangement, with four replications of eight birds each. Calcium and alkaline phosphatase (AP) enzyme concentrations in serum did not differ at 7, 14 and 21 days old. Serum phosphorus levels differed at 21 days and were higher with 12.5% L-Glu. Liver weight was higher in birds fed diet with 6.25% L-Glu and 12.5% L-Glu + 0.02 mg vit. K/kg. Total serum protein remained far below normal levels. Concentrations of the alanine aminotransferase enzyme were high in birds fed L-Glu and vit. K in the evaluated levels. Serum concentrations of alanine aminotransferase enzyme was much higher than that levels considered normal for birds fed diets containing L-Glu and vit. K, in the evaluated levels. Vitamin K levels affected nor the absorption of calcium and phosphorus and the AP enzyme neither the bone tissue metabolism. No-specific nitrogen level (glutamic acid) of 6.25% of the diets changed the metabolism of bone tissue, as a result of the higher concentration of alkaline phosphatase enzyme in the serum of birds.

  12. Characterization of the Human LPIN1-encoded Phosphatidate Phosphatase Isoforms*

    OpenAIRE

    Han, Gil-Soo; CARMAN, George M.

    2010-01-01

    The human LPIN1 gene encodes the protein lipin 1, which possesses phosphatidate (PA) phosphatase (3-sn-phosphatidate phosphohydrolase; EC 3.1.3.4) activity (Han, G.-S., Wu, W.-I., and Carman, G. M. (2006) J. Biol. Chem. 281, 9210–9218). In this work, we characterized human lipin 1 ?, ?, and ? isoforms that were expressed in Escherichia coli and purified to near homogeneity. PA phosphatase activities of the ?, ?, and ? isoforms were dependent on Mg2+ or Mn2+ ions at pH 7.5 at 37 °C. The activi...

  13. Downregulation of protein tyrosine phosphatase PTP-BL represses adipogenesis.

    OpenAIRE

    Glondu-Lassis, Murielle; Dromard, Mathilde; Chavey, Carine; Puech, Carole; Fajas, Lluis; Hendriks, Wiljan; Freiss, Gilles

    2009-01-01

    The insulin/insulin-like growth factor 1 (IGF-1) signaling pathway is a major regulator of adipose tissue growth and differentiation. We recently demonstrated that human protein tyrosine phosphatase (PTP) L1, a large cytoplasmic phosphatase also known as PTP-BAS/PTPN13/PTP-1E, is a negative regulator of IGF-1R/IRS-1/Akt pathway in breast cancer cells. This triggered us to investigate the potential role of PTPL1 in adipogenesis. To evaluate the implication of PTP-BL, the mouse orthologue of PT...

  14. Structure determination of T cell protein-tyrosine phosphatase

    DEFF Research Database (Denmark)

    Iversen, L.F.; Møller, K. B.; Pedersen, A.K.; Peters, Günther H.J.; Petersen, A.S.; Andersen, H.S.; Branner, S.; Mortensen, S.B.; Møller, N.P.H.

    2002-01-01

    Protein-tyrosine phosphatase 1B (PTP1B) has recently received much attention as a potential drug target in type 2 diabetes. This has in particular been spurred by the finding that PTP1B knockout mice show increased insulin sensitivity and resistance to diet-induced obesity. Surprisingly, the highly homologous T cell protein-tyrosine phosphatase (TC-PTP) has received much less attention, and no x-ray structure has been provided. We have previously co-crystallized PTP1B with a number of low molecu...

  15. Catalytic oxidation of soot over alkaline niobates

    International Nuclear Information System (INIS)

    Highlights: ? No previous reported studies about alkaline niobates as catalysts for soot oxidation. ? NaNbO3 and KNbO3 perovskite-type oxides show lower activation energy than other lanthanoid perovskite-type oxides. ? The alkaline niobate does not show deactivation by metal loss. - Abstract: The lack of studies in the current literature about the assessment of alkaline niobates as catalysts for soot oxidation has motivated this research. In this study, the synthesis, characterization and assessment of alkaline metal niobates as catalysts for soot combustion are reported. The solids MNbO3 (M = Li, Na, K, Rb) are synthesized by a citrate method, calcined at 450 °C, 550 °C, 650 °C, 750 °C, and characterized by AAS, N2 adsorption, XRD, O2-TPD, FTIR and SEM. All the alkaline niobates show catalytic activity for soot combustion, and the activity depends basically on the nature of the alkaline metal and the calcination temperature. The highest catalytic activity, expressed as the temperature at which combustion of carbon black occurs at the maximum rate, is shown by KNbO3 calcined at 650 °C. At this calcination temperature, the catalytic activity follows an order dependent on the atomic number, namely: KNbO3 > NaNbO3 > LiNbO3. The RbNbO3 solid do not follow this trend presumably due to the perovskite structure was not reached. The highest catalytic activity shown by of KNbO3, despite the lower apparent activation energy of NaNbO3, stress the importance of the metal nature and suggests the hypothesis that K+ ions are the active sites for soot combustion. It must be pointed out that alkaline niobate subjected to consecutive soot combustion cycles does not show deactivation by metal loss, due to the stabilization of the alkaline metal inside the perovskite structure.

  16. Increased tartrate-resistant acid phosphatase (TRAP expression in malignant breast, ovarian and melanoma tissue: an investigational study

    Directory of Open Access Journals (Sweden)

    Eck M

    2006-07-01

    Full Text Available Abstract Background Tartrate-resistant acid phosphatase (TRAP is a metalloprotein enzyme that belongs to the acid phosphatases and is known to be expressed by osteoclasts. It has already been investigated as a marker of bone metastases in cancer patients. In this study, which examined the value of serum TRAP concentrations as a marker of bone disease in breast cancer patients, we observed high concentrations of TRAP even in patients without bone metastases. To elucidate this phenomenon, we examined the expression of TRAP in breast cancer cells and the cells of several other malignancies. Methods TRAP concentrations in the serum of tumor patients were determined by ELISA. The expression of TRAP in breast, ovarian, and cervical cancer and malignant melanoma was analyzed by immunohistochemistry. RT-PCR and immunocytology were used to evaluate TRAP expression in cultured tumor cells. Results A marked increase in serum TRAP concentrations was observed in patients with breast and ovarian cancer, regardless of the presence or absence of bone disease. TRAP expression was found in breast and ovarian cancers and malignant melanoma, while cervical cancer showed only minimal expression of TRAP. Expression of TRAP was absent in benign tissue or was much less marked than in the corresponding malignant tissue. TRAP expression was also demonstrated in cultured primary cancer cells and in commercially available cell lines. Conclusion Overexpression of TRAP was detected in the cells of various different tumors. TRAP might be useful as a marker of progression of malignant disease. It could also be a potential target for future cancer therapies.

  17. The informative value of the radioimmunological determination of acid prostate phosphatase for the diagnostic of prostate carcinoma: A comparison of various reagent kits

    International Nuclear Information System (INIS)

    Patients of the university urological clinic in Freiburg gave, in all, 89 serum samples for the determination of acid prostate phosphatase using four different methods (3 radioimmunological methods and 1 enzymatic procedure) in order to compare the informative ability of these procedures in the diagnostic of prostate carcinoma. The results of the radioimmunological determinations agreed mostly with one another. They showed a higher sensitivity than the enzymatic method, but a lower specificity. The rate of false positive as well as false negative results is with both procedures too large, so that a reliable early diagnosis is not possible. Not until after metastasis of the carcinoma do all four methods give clearly pathological values. The acid prostate phosphatase determination is therefore suited for progress and therapy control. The values sink with successful therapy, rise again with recitivism. (TRV)

  18. Tyrosine Phosphatase Epsilon Is a Positive Regulator of Osteoclast Function in Vitro and In Vivo

    OpenAIRE

    Chiusaroli, Riccardo; Knobler, Hilla; Luxenburg, Chen; Sanjay, Archana; Granot-Attas, Shira; Tiran, Zohar; Miyazaki, Tsuyoshi; Harmelin, Alon; Baron, Roland; Elson, Ari

    2004-01-01

    Protein tyrosine phosphorylation is a major regulator of bone metabolism. Tyrosine phosphatases participate in regulating phosphorylation, but roles of specific phosphatases in bone metabolism are largely unknown. We demonstrate that young (

  19. Serum free hemoglobin test

    Science.gov (United States)

    Blood hemoglobin; Serum hemoglobin ... Hemoglobin (Hb) is the main component of red blood cells. It is a protein that carries oxygen. ... people may contain up to 5 mg/dL hemoglobin. Normal value ranges may vary slightly among different ...

  20. Biocatalysis with Sol-Gel Encapsulated Acid Phosphatase

    Science.gov (United States)

    Kulkarni, Suhasini; Tran, Vu; Ho, Maggie K.-M.; Phan, Chieu; Chin, Elizabeth; Wemmer, Zeke; Sommerhalter, Monika

    2010-01-01

    This experiment was performed in an upper-level undergraduate biochemistry laboratory course. Students learned how to immobilize an enzyme in a sol-gel matrix and how to perform and evaluate enzyme-activity measurements. The enzyme acid phosphatase (APase) from wheat germ was encapsulated in sol-gel beads that were prepared from the precursor…

  1. Impact of ionic aluminium on extracellular phosphatases in acidified lakes.

    Czech Academy of Sciences Publication Activity Database

    Bittl, T.; Vrba, Jaroslav; Nedoma, Ji?í; Kopá?ek, Ji?í

    2001-01-01

    Ro?. 3, ?. 9 (2001), s. 578-587. ISSN 1462-2912 R&D Projects: GA ?R GA206/97/0072; GA ?R GA206/00/0063 Keywords : acid phosphatases * pH effect * inhibition Subject RIV: DA - Hydrology ; Limnology Impact factor: 3.276, year: 2001

  2. Protein tyrosine phosphatases ? and ? perform nonredundant roles in osteoclasts

    OpenAIRE

    Finkelshtein, Eynat; Lotinun, Sutada; Levy-Apter, Einat; Arman, Esther; den Hertog, Jeroen; Baron, Roland; Elson, Ari

    2014-01-01

    The closely related tyrosine phosphatases PTPa and PTPe fulfill distinct roles in osteoclasts. The various effects of each PTP on podosome organization in osteoclasts are caused by their distinct N-termini. The function of PTPe in these cells requires the presence of its 12 N-terminal residues, in particular serine 2.

  3. Cytochemical localization of acid phosphatase in Stigeoclonium tenue (Chaetophorales, Chlorophyceae

    Directory of Open Access Journals (Sweden)

    Karina M. Michetti

    2006-12-01

    Full Text Available Nonspecific acid phosphatases are a group of enzymes whose activity increases the availability of exogenous and endogenous orthophosphate either through extra- or intracellular hydrolysis of phosphate compounds. Our study demonstrates the activity of acid phosphatases in the filamentous freshwater alga Stigeoclonium tenue. These enzymes were detected following a cerium-based method in which cerium was used as an orthophosphate-capture reagent. In thalli from S. tenue from the natural environment, acid phosphatases were found in the longitudinal cell wall, plasmalemma, and vacuole. In thalli from Bold's Basal Medium culture, these enzymes were found mainly in the plasmalemma; they were scarce in the cell wall. In the thalli grown in phosphate-enriched culture medium, enzymes were found only in the plasmalemma. The low availability of orthophosphate in the medium seems to induce the transport of these enzymes to the cell wall. Its abundance, on the contrary, seems to attenuate this response without affecting the localization of acid phosphatases in the plasmalemma.

  4. Cytochemical localization of acid phosphatase in Stigeoclonium tenue (Chaetophorales, Chlorophyceae)

    Scientific Electronic Library Online (English)

    Karina M., Michetti; Patricia I., Leonardi; Eduardo J., Cáceres.

    2006-12-01

    Full Text Available Nonspecific acid phosphatases are a group of enzymes whose activity increases the availability of exogenous and endogenous orthophosphate either through extra- or intracellular hydrolysis of phosphate compounds. Our study demonstrates the activity of acid phosphatases in the filamentous freshwater a [...] lga Stigeoclonium tenue. These enzymes were detected following a cerium-based method in which cerium was used as an orthophosphate-capture reagent. In thalli from S. tenue from the natural environment, acid phosphatases were found in the longitudinal cell wall, plasmalemma, and vacuole. In thalli from Bold's Basal Medium culture, these enzymes were found mainly in the plasmalemma; they were scarce in the cell wall. In the thalli grown in phosphate-enriched culture medium, enzymes were found only in the plasmalemma. The low availability of orthophosphate in the medium seems to induce the transport of these enzymes to the cell wall. Its abundance, on the contrary, seems to attenuate this response without affecting the localization of acid phosphatases in the plasmalemma.

  5. Par-4: A New Activator of Myosin Phosphatase

    OpenAIRE

    Vetterkind, Susanne; Lee, EunHee; Sundberg, Eric; Poythress, Ransom H.; Tao, Terence C.; Preuss, Ute; Morgan, Kathleen G

    2010-01-01

    We show here for the first time that the pro-apoptotic protein Par-4 binds to and activates myosin phosphatase (MP). During agonist stimulation, Par-4 facilitates ZIPK targeting and inhibitory phosphorylation of MP, however, phosphorylation of Par-4 is required for MP inhibition. Our model presents Par-4 as an amplifier of the MP activity range.

  6. Protein Tyrosine Phosphatases in Hypothalamic Insulin and Leptin Signaling.

    Science.gov (United States)

    Zhang, Zhong-Yin; Dodd, Garron T; Tiganis, Tony

    2015-10-01

    The hypothalamus is critical to the coordination of energy balance and glucose homeostasis. It responds to peripheral factors, such as insulin and leptin, that convey to the brain the degree of adiposity and the metabolic status of the organism. The development of leptin and insulin resistance in hypothalamic neurons appears to have a key role in the exacerbation of diet-induced obesity. In rodents, this has been attributed partly to the increased expression of the tyrosine phosphatases Protein Tyrosine Phosphatase 1B (PTP1B) and T cell protein tyrosine phosphatase (TCPTP), which attenuate leptin and insulin signaling. Deficiencies in PTP1B and TCPTP in the brain, or specific neurons, promote insulin and leptin signaling and prevent diet-induced obesity, type 2 diabetes mellitus (T2DM), and fatty liver disease. Although targeting phosphatases and hypothalamic circuits remains challenging, recent advances indicate that such hurdles might be overcome. Here, we focus on the roles of PTP1B and TCPTP in insulin and leptin signaling and explore their potential as therapeutic targets. PMID:26435211

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

    Scientific Electronic Library Online (English)

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

    2014-08-01

    Full Text Available We evaluated the concentrations of 25-hydroxyvitamin D [25(OH)D] in children and adolescents with juvenile systemic lupus erythematosus (JSLE) and associated them with disease duration and activity, use of medication (chloroquine and glucocorticoids), vitamin D intake, calcium and alkaline phosphata [...] se levels, and bone mineral density. Thirty patients with JSLE were evaluated and compared to 30 healthy individuals, who were age and gender matched. Assessment was performed of clinical status, disease activity, anthropometry, laboratory markers, and bone mineral density. The 30 patients included 25 (83.3%) females and 16 (53.3%) Caucasians, with a mean age of 13.7 years. The mean age at diagnosis was 10.5 years and mean disease duration was 3.4 years. Mean levels of calcium, albumin, and alkaline phosphatase were significantly lower in patients with JSLE compared with controls (P

  8. Effects of Dietary Furundu (Fermented Hibiscus sabdariffa Seed Supplementation on the Performance, Some Blood and Serum Parameters and Histopathology of Wistar Rats

    Directory of Open Access Journals (Sweden)

    Warda S. Abdelgadir

    2012-04-01

    Full Text Available The present study was conducted to obtain information on the effects of various dietary doses of the fermented Hibiscus sabdariffa seed (Furundu on Wistar rats. Emphasis was put on changes on growth, lesions, alteration in hematology and serobiochemical constituents and histopathology of treated rats. Fermented Hibiscus sabdariffa seed product (Furundu was fed to Wistar rats at 10, 15 and 20%, respectively of the standard diet for 12 weeks. Incorporation in the diet of the three doses of both was toxic but not fatal. Depression in growth and hepatonephrotoxicity were observed in rats that had been given 10, 15 and 20% Furundu, respectively for 12 weeks. These findings were accompanied by leukocytosis, macrocytic normochromic anaemia in rats fed on 20% Furundu and macrocytic hypochromic, normocytic hypochromic anaemia in those fed on 10 and 15% Furundu respectively, in addition to alterations in serum aspartate aminotransferase (AST, alanine aminotransferase (ALT and alkaline phosphatase (ALP activity and albumin, globulin, bilirubin and cholesterol concentrations. At the end of the experimental period (12 weeks, the total protein of all the test groups and the globulin concentration of group 2 fed on 15% Furundu decreased significantly (p<0.05, albumin concentration of all the test groups and the urea concentration of group 3 fed on 15% Furundu remained unchanged. Histopathological examination showed Fatty cytoplasmic vacuolation of the centrilobular hepatocytes and lymphocytic infiltration, desquamation of the intestinal epithelium and intestinal congestion were noticed in rats fed on 20% Furundu. Fermented Hibiscus sabdariffa seed product Furundu is considered enterohepatonephrotoxic to Wistar rats at dietary levels of 10, 15 and 20%, respectively.

  9. Effect of Turmeric and Carrot Seed Extracts on Serum Liver Biomarkers and Hepatic Lipid Peroxidation, Antioxidant Enzymes and Total Antioxidant Status in Rats

    Directory of Open Access Journals (Sweden)

    Adel Rezaei-Moghadam

    2012-07-01

    Full Text Available Introduction: Pathogenic role of free radicals are well known in various metabolic diseases. They originate from internal and external sources of body. Essential roles of antioxidant defense system for cellular redox regulation and free radical scavenging activity were described in this study. Many in vitro investigations have shown that turmeric (TE and carrot seed extract (CSE exhibits to possess antioxidant activities. In this study, we evaluated the antioxidant potentials of ethanolic TE and CSE based on in vivo experiment in the rats. Methods: Animals were assigned to six groups: the 1st and 2nd groups were control groups and 2nd group received 0.2 ml dimethyl sulphoxide as vehicle treated group; other four experimental groups received different doses of TE (100, 200 mg/kg b.w. and CSE (200, 400 mg/kg b.w. by gavages, respectively for a period of one month. The indicators of oxidative stress, lipids peroxidation, markers of hepatocyte injury and biliary function markers were measured. Results: The levels of superoxide dismutase, catalase, and glutathione peroxidase were significantly stimulated in the hepatic tissue of treatment groups. The malondialdehyde contents of liver tissue were significantly reduced in the groups fed with TE and CSE. Serum levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase, in treated groups were found to be significantly decreased, whereas albumin and total protein increased as compared to the control groups (P<0.05. Conclusion: this study showed that the regular intake of TE and CSE through the diet can improve antioxidant status and inhibit peroxidation activity in the liver tissue so that using these extracts may protect tissue oxidative stress.

  10. Ferritin in cattle serum

    International Nuclear Information System (INIS)

    A commercially available radioimmunoassay kit for human serum ferritin was used to determine the ferritin concentration in serum or plasma of 41 calves from 0 to 106 days of age, 192 cows and 35 bulls from 2 to 11 years of age. The geometric average concentration of ferritin was 2.1, 12.6 and 4.5 ng/ml for the calves, cows and bulls, respectively. The cows were statistically different from the calves and bulls; there were no differences between the calves and bulls. Within the cows one herd was found to have lower serum ferritin levels than all the other herds (P < 0.05) but no differences in packed cell volumes were present. The data suggest that a radioimmunoassay procedure with a ferritin antibody specific for bovine ferritin could be useful in the study of iron metabolism in cattle. (author)

  11. Some durability aspects of hybrid alkaline cements

    Directory of Open Access Journals (Sweden)

    Donatello S.

    2014-04-01

    Full Text Available Blended cements that contain a high content of fly ash and a low content of Portland cement typically suffer from low early strength development and long setting times. Recently, one method of overcoming these problems has been to use an alkali activator to enhance the reactivity of fly ash particles at early ages. Such cements can be grouped under the generic term “hybrid alkaline cements”, where both cement clinker and fly ash, encouraged by the presence of alkalis, are expected to contribute to cementitious gel formation. The work presented here examines some of the durability aspects of high fly ash content hybrid alkaline cement. Specifically, the aspects investigated were: exposure at high temperatures (up to 1000°C, resistance to immersion in aggressive solutions and susceptibility to the alkali aggregate reaction. All tests were repeated with a commercially available sulfate resistant Portland cement for comparison. When exposed to high temperatures, the hybrid alkaline cement showed strikingly different behaviour compared to the control Portland cement, showing fewer micro-cracks and maintaining residual compressive strengths at least equal to original strengths. Beyond 700°C, the hybrid alkaline cement began to sinter, which resulted in shrinkage of around 5% and a 100% increase in residual compressive strengths. No such sintering event was noted in the control Portland cement, which showed a drastic loss in residual compressive strengths upon heating. In immersion tests, the hybrid alkaline cement possessed excellent resistance to sulfate and seawater attack, similar to the control sulfate resistant cement. Both cements were however severely degraded by immersion in 0.1M HCl for 90 days. Both binders complied with the accelerated alkali-aggregate test but when this test was extended, the hybrid alkaline binder showed much greater dimensional stability. Possible reasons for the differences in durability behaviour in both cements are discussed, based on experimental evidence provided.

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

    DEFF Research Database (Denmark)

    Zhang, Yao; Cheng, Yajun; Hansen, Gert H; Niels-Christiansen, Lise-Lotte; Koentgen, Frank; Ohlsson, Lena; Nilsson, Ake; Duan, Rui-Dong

    2011-01-01

    Alkaline sphingomyelinase (alk-SMase) hydrolyses sphingomyelin (SM) to ceramide in the gut. To evaluate the physiological importance of the enzyme, we generated alk-SMase knockout (KO) mice by the Cre-recombinase-Locus of X-over P1(Cre-LoxP) system and studied SM digestion. Both wild-type (WT) and KO mice were fed ³H-palmitic acid labeled SM together with milk SM by gavage. The lipids in intestinal content, intestinal tissues, serum, and liver were analyzed by TLC. In KO mice, nondigested ³H-SM ...

  13. Electrochemical behaviour of alkaline copper complexes

    Indian Academy of Sciences (India)

    C L Aravinda; S M Mayanna; V R Muralidharan

    2000-10-01

    A search for non-cyanide plating baths for copper resulted in the development of alkaline copper complex baths containing trisodium citrate [TSC] and triethanolamine [TEA]. Voltammetric studies were carried out on platinum to understand the electrochemical behaviour of these complexes. In TSC solutions, the deposition of copper involves the slow formation of a monovalent species. Adsorption of this species obeys Langmuir isotherm. In TEA solutions the deposition involves the formation of monovalent ions obeying the non-activated Temkin isotherm. Conversion of divalent to monovalent copper is also slow. In TEA and TSC alkaline copper solutions, the predominant species that undergo stepwise reduction contain only TEA ligands

  14. [Postoperative alkaline reflux gastritis (author's transl)].

    Science.gov (United States)

    Rothmund, M; Neher, M

    1977-02-25

    Postoperative alkaline reflux gastritis is a distinct clinical entity occuring after operations enlarging, bypassing or resecting the pylorus. Reflux of alkaline duodenal content into the stomach is the causative factor. Primarily bile acids have an aggressive effect and lead to a destruction of the gastric mucosal barrier. Epigastric pain, fullness after meals and bile vomiting are the main symptoms. Gastroscopy with biopsy reveals a severe chronic atrophic gastritis and bile reflux. In most cases an achlorhydria that can be histamin-resistant is present. For adequate treatment surgical procedures diverting the bile flow from the stomach should be performed. PMID:836529

  15. Alkaline earth filled nickel skutterudite antimonide thermoelectrics

    Science.gov (United States)

    Singh, David Joseph

    2013-07-16

    A thermoelectric material including a body centered cubic filled skutterudite having the formula A.sub.xFe.sub.yNi.sub.zSb.sub.12, where A is an alkaline earth element, x is no more than approximately 1.0, and the sum of y and z is approximately equal to 4.0. The alkaline earth element includes guest atoms selected from the group consisting of Be, Mb, Ca, Sr, Ba, Ra and combinations thereof. The filled skutterudite is shown to have properties suitable for a wide variety of thermoelectric applications.

  16. Cambios séricos en las enzimas alt, ast, fa, ck-total y ldh inducidos por veneno de crotalus durissus cumanensis en ratones balb/c / Serum Changes in alt, ast, fa, ck-total and ldh Enzymes Induced by the Venom of Crotalus durissus cumanensis Snake in Mouse Balb/C

    Scientific Electronic Library Online (English)

    José, Noriega-Alvarado; David, Colmenarez; Alexander, Mogollón; Nelson, Márquez; Virgilio, Hernández; Mirleny, Pérez.

    2009-08-01

    Full Text Available El objetivo del presente trabajo fue medir los cambios séricos en las enzimas ALT, AST, FA, CK-TOTAL Y LDH inducidos por veneno de Crotalus durissus cumanensis en ratones balb/c que fueron inoculados con veneno crudo a una dosis de 0,75 mg proteínas/kg vía intraperitoneal, y los controles con soluci [...] ón fisiológica. Las muestras fueron obtenidas a diferentes intervalos de tiempo post inyección (1; 3; 6; 12 y 24 h) y se determinó la actividad de las enzimas: Alanina Aminotransferasa (ALT), Aspartato Aminotransferasa (AST), Fosfatasa Alcalina (FA), Creatina cinasa-total (CK-total) y Lactato Deshidrogenasa (LDH). Se observó diferencia significativa (P Abstract in english The objective of the present work was to measure the serum changes in the enzymes ALT, AST, AF, TOTAL-CK and LDH induced by the venom of Crotalus durissus cumanensis in Balb/C mice which been inoculated with crude venom in a doses of 0.75 mg/kg by intraperitoneal way, and the control with physiologi [...] cal solution. The samples were obtained at different intervals post-injection (1, 3, 6, 12 and 24 h) and was determinated the activity of the enzymes: Alanine Aminotransferase (ALT), Aspatate Aminotransferase (AST) and Alkaline Phosphatase (AF); Total Creatin Kinase (TCK) and Lactate Deshidrogenase (LDH). There was significant difference (P

  17. The Alkaline Diet: Is There Evidence That an Alkaline ph Diet Benefits Health?

    International Nuclear Information System (INIS)

    This review looks at the role of an alkaline diet in health. Pub med was searched looking for articles on ph, potential renal acid loads, bone health, muscle, growth hormone, back pain, vitamin D and chemotherapy. Many books written in the lay literature on the alkaline diet were also reviewed and evaluated in light of the published medical literature. There may be some value in considering an alkaline diet in reducing morbidity and mortality from chronic diseases and further studies are warranted in this area of medicine

  18. Protein tyrosine phosphatases in the JAK/STAT pathway

    OpenAIRE

    Xu, Dan; Qu, Cheng-Kui

    2008-01-01

    The Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway is crucial in controlling cellular activities in response to extracellular cytokines. Dysfunctions of the JAK/STAT pathway result in various hematopoietic and immune disorders. The central events in regulating this pathway are tyrosine phosphorylation and dephosphorylation of the signaling components, which are carried out by protein tyrosine kinases and protein tyrosine phosphatases (PTP), respectively. He...

  19. MAP Kinase Phosphatase-1 Protects against Inflammatory Bone Loss

    OpenAIRE

    R SARTORI; Li, F.; Kirkwood, K.L.

    2009-01-01

    The mitogen-activated protein (MAP) kinase phosphatase (MKP) family plays an important function in regulating the pro-inflammatory cytokines by de-activating MAP kinases. MKP-1 is essential for the dephosphorylation of p38 MAP kinase that regulates expression of IL-6, TNF-?, and IL-1?. We hypothesized that MKP-1 regulates inflammatory bone loss in experimental periodontitis. Wild-type and Mkp-1?/? mice received A. actinomycetemcomitans LPS injection in the palatal region or PBS control 3 time...

  20. Light availability may control extracellular phosphatase production in turbid environments.

    Czech Academy of Sciences Publication Activity Database

    Rychtecký, Pavel; ?eháková, Klára; Kozlíková, Eliška; Vrba, Jaroslav

    2015-01-01

    Ro?. 69, ?. 1 (2015), s. 37-44. ISSN 0095-3628 R&D Projects: GA ?R(CZ) GA206/09/0309; GA ?R(CZ) GAP504/11/2177; GA ?R(CZ) GAP504/11/2182 Institutional support: RVO:60077344 Keywords : phytoplankton * phosphatase activity * ELF97 phosphate Subject RIV: DA - Hydrology ; Limnology Impact factor: 2.973, year: 2014

  1. LAFORIN: A DUAL SPECIFICITY PHOSPHATASE THAT DEPHOSPHORYLATES COMPLEX CARBOHYDRATES

    OpenAIRE

    Worby, Carolyn A.; Gentry, Matthew S.; Dixon, Jack E

    2006-01-01

    Laforin is the only phosphatase in the animal kingdom that contains a carbohydrate-binding module (CBM). Mutations in the gene encoding laforin result in Lafora disease (LD), a fatal autosomal recessive neurodegenerative disorder, which is diagnosed by the presence of intracellular deposits of insoluble complex carbohydrates known as Lafora bodies (LB). We demonstrate that laforin interacts with proteins known to be involved in glycogen metabolism and rule out several of these proteins as pot...

  2. Bod1 regulates protein phosphatase 2A at mitotic kinetochores

    OpenAIRE

    Porter, Iain M.; Schleicher, Katharina; Porter, Michael; SWEDLOW, JASON R

    2013-01-01

    Mitotic entry and progression require the activation of several mitotic kinases and the proper regulation and localization of several phosphatases. The activity and localization of each of these enzymes is tightly controlled through a series of specific activators, inhibitors and regulatory subunits. Two proteins, Ensa and Arpp-19, were recently identified as specific inhibitors of PP2A-B55 and are critical for allowing full activity of Cdk1/cyclin B and entry into mitosis. Here we show that ...

  3. Protein phosphatase 1 and memory: practice makes PP1 imperfect?

    OpenAIRE

    Waddell, S.

    2003-01-01

    Long-lasting memories are most efficiently formed by multiple training sessions separated by appropriately timed intervals. A recent study revealed that expression of a transgene encoding an inhibitor of protein phosphatase 1 (PP1) in the forebrain enhanced memory formed during sub-optimal training. Thus, PP1 apparently constrains memory formation in the mouse. Furthermore, the report proposes that PP1 promotes forgetting.

  4. Colorimetric Determination of Pure Mg2+-dependent Phosphatidate Phosphatase Activity

    OpenAIRE

    Havriluk, Tara; Lozy, Fred; Siniossoglou, Symeon; Carman, George M.

    2007-01-01

    The malachite green-molybdate reagent was used for a colorimetric assay of pure Mg2+-dependent phosphatidate phosphatase activity. This enzyme plays a major role in fat metabolism. Enzyme activity was linear with time and protein concentration, and with the concentration of water-soluble dioctanoyl phosphatidate. The colorimetric assay was used to examine enzyme inhibition by phenylglyoxal, propranolol, and dimethyl sulfoxide. Pure enzyme and a water-soluble phosphatidate substrate were requi...

  5. Searching for the role of protein phosphatases in eukaryotic microorganisms

    Directory of Open Access Journals (Sweden)

    da-Silva A.M.

    1999-01-01

    Full Text Available Preference for specific protein substrates together with differential sensitivity to activators and inhibitors has allowed classification of serine/threonine protein phosphatases (PPs into four major types designated types 1, 2A, 2B and 2C (PP1, PP2A, PP2B and PP2C, respectively. Comparison of sequences within their catalytic domains has indicated that PP1, PP2A and PP2B are members of the same gene family named PPP. On the other hand, the type 2C enzyme does not share sequence homology with the PPP members and thus represents another gene family, known as PPM. In this report we briefly summarize some of our studies about the role of serine/threonine phosphatases in growth and differentiation of three different eukaryotic models: Blastocladiella emersonii, Neurospora crassa and Dictyostelium discoideum. Our observations suggest that PP2C is the major phosphatase responsible for dephosphorylation of amidotransferase, an enzyme that controls cell wall synthesis during Blastocladiella emersonii zoospore germination. We also report the existence of a novel acid- and thermo-stable protein purified from Neurospora crassa mycelia, which specifically inhibits the PP1 activity of this fungus and mammals. Finally, we comment on our recent results demonstrating that Dictyostelium discoideum expresses a gene that codes for PP1, although this activity has never been demonstrated biochemically in this organism.

  6. Searching for the role of protein phosphatases in eukaryotic microorganisms

    Scientific Electronic Library Online (English)

    A.M., da-Silva; P.D.A., Zapella; L.P.M., Andrioli; R.B., Campanhã; L.C., Fiorini; L.C., Etchebehere; J.C., da-Costa-Maia; H.F., Terenzi.

    1999-07-01

    Full Text Available Preference for specific protein substrates together with differential sensitivity to activators and inhibitors has allowed classification of serine/threonine protein phosphatases (PPs) into four major types designated types 1, 2A, 2B and 2C (PP1, PP2A, PP2B and PP2C, respectively). Comparison of seq [...] uences within their catalytic domains has indicated that PP1, PP2A and PP2B are members of the same gene family named PPP. On the other hand, the type 2C enzyme does not share sequence homology with the PPP members and thus represents another gene family, known as PPM. In this report we briefly summarize some of our studies about the role of serine/threonine phosphatases in growth and differentiation of three different eukaryotic models: Blastocladiella emersonii, Neurospora crassa and Dictyostelium discoideum. Our observations suggest that PP2C is the major phosphatase responsible for dephosphorylation of amidotransferase, an enzyme that controls cell wall synthesis during Blastocladiella emersonii zoospore germination. We also report the existence of a novel acid- and thermo-stable protein purified from Neurospora crassa mycelia, which specifically inhibits the PP1 activity of this fungus and mammals. Finally, we comment on our recent results demonstrating that Dictyostelium discoideum expresses a gene that codes for PP1, although this activity has never been demonstrated biochemically in this organism.

  7. Acid phosphatases in seeds and developing of squash (Cucurbita ficifolia

    Directory of Open Access Journals (Sweden)

    Irena Lorenc-Kubis

    2014-02-01

    Full Text Available Changes in protein content and acid phosphatase activity were followed during germination (imbition through seedlings development in extracts from cotyledons of squash (Cucurbita ficifolia. It has been shown that the activity of acid phosphatase was initially low and than increased to a maximum after 6 days of imbition. Acid phosphates were isolated from cotyledons of seeds and from 6-, 10- and 22-days old seedlings by extraction the proteins with 0.1 M acetate buffer pH 5.1, precipitation with ethanol and by affinity chromatography on con A-Sepharose. Two glycoprotein enzymes AcPase Ba and AcPase Bb which differ in their affinity to immobilized con A were obtained. Both acid phosphatates retained the enzyme activity after binding to free con A. Rocket affinity electrophoresis of AcPase Ba and AcPase Bb, isolated from cotyledons of seeds and seedlings, revealed differences in their ability to bind to con A during seeds germination and seedling develop-ment indicating changes in their sugar component. Con A was found to activate both enzymes. The enzymes cross-reacted with monospecific antibodies raised against grass seed acid phosphatate Ba indicating an antigenic relationship between squash and grass acid phosphatases.

  8. Displacement affinity chromatography of protein phosphatase one (PP1 complexes

    Directory of Open Access Journals (Sweden)

    Gourlay Robert

    2008-11-01

    Full Text Available Abstract Background Protein phosphatase one (PP1 is a ubiquitously expressed, highly conserved protein phosphatase that dephosphorylates target protein serine and threonine residues. PP1 is localized to its site of action by interacting with targeting or regulatory proteins, a majority of which contains a primary docking site referred to as the RVXF/W motif. Results We demonstrate that a peptide based on the RVXF/W motif can effectively displace PP1 bound proteins from PP1 retained on the phosphatase affinity matrix microcystin-Sepharose. Subsequent co-immunoprecipitation experiments confirmed that each identified binding protein was either a direct PP1 interactor or was in a complex that contains PP1. Our results have linked PP1 to numerous new nuclear functions and proteins, including Ki-67, Rif-1, topoisomerase II?, several nuclear helicases, NUP153 and the TRRAP complex. Conclusion This modification of the microcystin-Sepharose technique offers an effective means of purifying novel PP1 regulatory subunits and associated proteins and provides a simple method to uncover a link between PP1 and additional cellular processes.

  9. Delignification of biomass using alkaline glycerol

    Energy Technology Data Exchange (ETDEWEB)

    Kucuk, M. [YYU Educational Facility, Van (Turkey)

    2005-10-15

    Ailanthus altissima and Spruce orientalis chips were delignified by using glycerol as a solvent and alkaline-gliycerol with and without catalyst at different temperatures (438, 458, 478, and 498 K) in atmospheric pressure, and results were compared with the other organic solvent systems. (Author)

  10. Biomass production on saline-alkaline soils

    Energy Technology Data Exchange (ETDEWEB)

    Chaturvedi, A.N.

    1985-01-01

    In a trial of twelve tree species (both nitrogen fixing and non-fixing) for fuel plantations on saline-alkaline soil derived from Gangetic alluvium silty clay, Leucaena leucocephala failed completely after showing rapid growth for six months. Results for other species at age two showed that Prosopis juliflora had the best productivity.

  11. Second Trimester Maternal Serum Screening

    Science.gov (United States)

    ... Home Visit Global Sites Search Help? Second Trimester Maternal Serum Screening Share this page: Was this page helpful? Also known as: AFP Maternal; Maternal Serum AFP; MSAFP; msAFP; Triple Screen; Triple ...

  12. CHANGES IN SERUM ENZYMES LEVELS ASSOCIATED WITH LIVER FUNCTIONS IN STRESSED MARWARI GOAT

    Directory of Open Access Journals (Sweden)

    Kataria N.

    2011-03-01

    Full Text Available Serum enzyme levels were determined in goats of Marwari breed belonging to farmers’ stock of arid tract of Rajasthan state, India. The animals were grouped into healthy and stressed comprising of gastrointestinal parasiticised, pneumonia affected, and drought affected. The serum enzymes determined were sorbitol dehydrogenase, malate dehydrogenase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, ornithine carbamoyl transferase, gamma-glutamayl transferase, 5’nucleotidase, glucose-6-phosphatase, arginase, and aldolase. In stressed group the mean values of all the enzymes increased significantly (p?0.05 as compared to respective healthy mean value. All the enzymes showed highest values in the gastrointestinal parasiticised animals and least values in the animals having pneumonia. In gastrointestinal parasiticised animals maximum change was observed in G-6-Pase activity and minimum change was observed in malate dehydrogenase mean value. It was concluded that Increased activity of all the serum enzymes was due to modulation of liver functions directly or indirectly.

  13. Seasonal changes in phosphatase activities in Toulon Bay (France).

    Science.gov (United States)

    Boge, Gérard; Jean, Natacha; Jamet, Jean-Louis; Jamet, Dominique; Richard, Simone

    2006-02-01

    We studied the characteristics of the phosphatase activity (Km and Vmax) in total seawater and in particulate material of the three main plankton classes (0.25-5, 5-90 and >90 microm) in a coastal marine ecosystem of Toulon Bay (French Mediterranean Sea). The measurement of the hydrolysis of sodium paranitrophenylphosphate (pNPP), a substrate of phosphatase, revealed low and high affinity components in unfiltered seawater and in particulate matter. In unfiltered seawater, the low affinity activity was predominant from October to March during phytoplankton development. The high affinity activity dominated from April to June and was significantly correlated with the bacterial abundances. The phosphatase behaviour in the particulate material differs from that in the unfiltered seawater. The activity of the three particulate classes was generally much lower than that of unfiltered seawater, particularly the low affinity activity. The >90 microm size fraction consisted in greater part of zooplankton. In this size class, the activity (nmol l(-1) h(-1)) of the low affinity component was predominant from May to August, when the abundance of the larvae of copepods (copepodites) was highest. Its high specific activity (Activity/Protein concentration as nmol l(-1) h(-1) microg(-1)) was particularly elevated during this period. The 5-90 microm fraction consisted of phytoplankton cells, especially Dinoflagellates. Between September and January, the activity (nmol l(-1) h(-1)) of this size class was mostly supported by the low affinity component. The specific activity (nmol l(-1) h(-1) microg(-1)) of the high affinity component was highest in June and August. No significant correlation was found between phosphatase activities and chlorophyll a or total cell abundance. In return temporary relationships with specific taxa exist in particular with Ceratium spp., Gymnodinium spp. and Protoperidinium spp. The contribution of the 0.25-5 microm size class exceeded rarely 20% of the total particulate activity. Between June and August, high specific activities (nmol l(-1) h(-1) microg(-1)) were observed for its high affinity component. In autumn, strong rainfall increased the phosphate and nitrate concentrations and led to a drop in salinity, which probably explains the low phosphatase activities (nmol l(-1) h(-1)) and cell densities observed during this period. PMID:15935465

  14. Some hereditary blood factors of the Bengali Muslim of Bangladesh (red cell enzymes, haemoglobins, and serum proteins).

    Science.gov (United States)

    Papiha, S S; Roberts, D F; Ali, S G; Islam, M M

    1975-08-25

    In a sample of Bengali Muslems from Dacca, haptoglobin, group-specific component, haemoglobin, adenosine deaminase, adenylate kinase, 6-phosphogluconate dehydrogenase, phosphoglucomutase, acid phosphatase and several other red cell enzyme types were studied. For most serum protein and red cell enzyme systems the gene frequencies are similar to those in other populations to the west of Bangladesh, but others suggest affinity with populations to the east. PMID:51817

  15. Dephosphorylation of chicken cardiac myofibril C-protein by protein phosphatases 1 and 2A

    International Nuclear Information System (INIS)

    C-Protein, which is a regulatory component of cardiac muscle myofibrils, is phosphorylated in response to ?-adrenergic agonists by a cAMP-dependent mechanism and dephosphorylated in response to cholinergic agonists. It is believed that the cAMP-dependent phosphorylation is due to cAMP-dependent protein kinase. The protein phosphatase(s) involved in the dephosphorylation of C-protein has not been determined. In this study, chicken cardiac C-protein was phosphorylated with the cAMP-dependent protein kinase to about 3 mol phosphate/mol C-protein. Incubation of [32P]C-protein with the catalytic subunit of protein phosphatase 1 or 2A rapidly removed 30-40% of 32[P]. Phosphopeptide maps and phosphoamino acid analysis revealed that the major site(s) dephosphorylated by either phosphatase was a phosphothreonine residue(s) located on the same tryptic peptide and on the same CNBr fragment. Increasing the incubation period or the phosphatase concentration did not result in any further dephosphorylation of C-protein by phosphatase 1, but phosphatase 2A completely dephosphorylated C-protein. Preliminary studies showed that the major protein phosphatase associated with the myofibril was phosphatase 2A. These results indicate the phosphatase 2A may be important in the regulation of the phosphorylation state of C-protein

  16. Tyr Phosphatase-Mediated P-ERK Inhibition Suppresses Senescence in EIA + v-raf Transformed Cells, Which, Paradoxically, Are Apoptosis-Protected in a MEK-Dependent Manner

    Directory of Open Access Journals (Sweden)

    Stefania De Vitis

    2011-02-01

    Full Text Available Activation of the Ras-Raf-extracellular signal-regulated kinase (ERK pathway causes not only proliferation and suppression of apoptosis but also the antioncogenic response of senescence. How these contrasting effects are reconciled to achieve cell transformation and cancer formation is poorly understood. In a system of two-step carcinogenesis (dedifferentiated PC EIA, transformed PC EIA-polyoma-middle T [PC EIA + Py] and PC EIA-v-raf [PC EIA + raf] cells], v-raf cooperated with EIA by virtue of a strong prosurvival effect, not elicited by Py-middle T, evident toward serum-deprivation-and H2O2-induced apoptosis. Apoptosis was detected by DNA fragmentation and annexin V staining. The prosurvival function of v-raf was, in part, mitogen-activated protein kinase/ERK kinase (MEK-dependent, as shown by pharmacological MEK inhibition. The MEK-dependent antiapoptotic effect of v-raf was exerted despite a lower level of P-ERK1/2 in EIA + raf cells with respect to EIA + Py/EIA cells, which was dependent on a high tyrosine phosphatase activity, as shown by orthovanadate blockade. An ERK1/2 tyrosine phosphatase was likely involved. The high tyrosine phosphatase activity was instrumental to the complete suppression of senescence, detected by ?-galactosidase activity, because tyrosine phosphatase blockade induced senescence in EIA + raf but not in EIA + Py cells. High tyrosine phosphatase activity and evasion from senescence were confirmed in an anaplastic thyroid cancer cell line. Therefore, besides EIA, EIA + raf cells suppress senescence through a new mechanism, namely, phosphatase-mediated P-ERK1/2 inhibition, but, paradoxically, retain the oncogenic effects of the Raf-ERK pathway. We propose that the survival effect of Raf is not a function of absolute P-ERK1/2 levels at a given time but is rather dynamically dependent on greater variations after an apoptotic stimulus.

  17. The cell wall-targeted purple acid phosphatase AtPAP25 is critical for acclimation of Arabidopsis thaliana to nutritional phosphorus deprivation.

    Science.gov (United States)

    Del Vecchio, Hernan A; Ying, Sheng; Park, Joonho; Knowles, Vicki L; Kanno, Satomi; Tanoi, Keitaro; She, Yi-Min; Plaxton, William C

    2014-11-01

    Plant purple acid phosphatases (PAPs) belong to a relatively large gene family whose individual functions are poorly understood. Three PAP isozymes that are up-regulated in the cell walls of phosphate (Pi)-starved (-Pi) Arabidopsis thaliana suspension cells were purified and identified by MS as AtPAP12 (At2g27190), AtPAP25 (At4g36350) and AtPAP26 (At5g34850). AtPAP12 and AtPAP26 were previously isolated from the culture medium of -Pi cell cultures, and shown to be secreted by roots of Arabidopsis seedlings to facilitate Pi scavenging from soil-localized organophosphates. AtPAP25 exists as a 55 kDa monomer containing complex NX(S/T) glycosylation motifs at Asn172, Asn367 and Asn424. Transcript profiling and immunoblotting with anti-AtPAP25 immune serum indicated that AtPAP25 is exclusively synthesized under -Pi conditions. Coupled with potent mixed-type inhibition of AtPAP25 by Pi (I50 = 50 ?m), this indicates a tight feedback control by Pi that prevents AtPAP25 from being synthesized or functioning as a phosphatase except when Pi levels are quite low. Promoter-GUS reporter assays revealed AtPAP25 expression in shoot vascular tissue of -Pi plants. Development of an atpap25 T-DNA insertion mutant was arrested during cultivation on soil lacking soluble Pi, but rescued upon Pi fertilization or complementation with AtPAP25. Transcript profiling by quantitative RT-PCR indicated that Pi starvation signaling was attenuated in the atpap25 mutant. AtPAP25 exhibited near-optimal phosphatase activity with several phosphoproteins and phosphoamino acids as substrates. We hypothesize that AtPAP25 plays a key signaling role during Pi deprivation by functioning as a phosphoprotein phosphatase rather than as a non-specific scavenger of Pi from extracellular P-monoesters. PMID:25270985

  18. Radioimmunoassay of serum ferritin

    International Nuclear Information System (INIS)

    Purified human spleen ferritin was labelled with 125I. On Sepharose 6-B gel filtration four species of labelled products were separated: a component with a higher molecular weight than ferritin; a component which is eluted in the same volume as unlabelled ferritin; and two labelled compounds with molecular weights lower than ferritin. When these labelled materials were used in a double antibody radioimmunoassay, the higher molecular weight fraction showed variable and high non-specific binding and was poorly displaced by unlabelled ferritin; the fraction behaving like true ferritin gave good standard curves and showed non-specific binding of less than 1%. The remaining two components showed poor binding to rabbit antiferritin. Using labelled material from the second fraction, a double antibody radioimmunoassay capable of measuring 2?g ferritin protein/litre of serum was developed. Inter-and intra-assay variation was between 3% and 8% over a concentration range of 0 to 250 ?g ferritin protein/litre. Good agreement between serum ferritin levels assayed by the present method and by an immunoradiometric method was obtained. Labelled ferritin was stable for at least six weeks. The simplicity of the methodology makes it possible to assay serum ferritin in large batches. (author)

  19. Radioimmunoassay for serum ferritin

    International Nuclear Information System (INIS)

    We describe the development and evaluation of a serum ferritin radioimmunoassay, in which 125I-labeled ferritin and rabbit anti-ferritin antibody are used. Goat anti-rabbit gamma-globulin antibody, together with polyethylene glycol, is used as the separating reagent. The assay has a working range up to 500 ?g of ferritin per litre, and a sample requirement of 75 ?l of serum for assay at two dilutions. The assay requires 24 h. It has a sensitivity of 1.5 ?g of ferritin per litre and a long-term precision (CV) of 13%. Reference intervals for a population of men were 18 to 330 ?g/litre, with no marked age dependence, while those for a population of women older than 50 years were 18 to 200 ?g/litre. Many apparently healthy women in the 20 to 50 year age group have much lower concentrations. Serum ferritin concentrations of <18 ?g/litre are indicative of iron deficiency, defined as the absence of stainable iron in an aspirate of bone marrow

  20. Alteraciones de analitos séricos y de orina en perros diabéticos: Informe de 30 casos / Alterations of serum and urine analytes in diabetic dogs: 30 cases report

    Scientific Electronic Library Online (English)

    Samuel Genaro, Jardón Herrera; Rosa Luz, Mondragón Vargas; Luis Enrique, García Ortuño; Jan, Bouda.

    2008-12-01

    Full Text Available El objetivo de este estudio fue describir alteraciones en la bioquímica clínica sanguínea y en el urianálisis, con el fin de establecer un diagnóstico integral en perros con diabetes mellitus. Se analizaron 30 casos de perros diabéticos. Los criterios de inclusión fueron: hiperglucemia superior a 14 [...] mmol/L, glucosuria, densidad urinaria superior a 1.014 y poliuria-polidipsia. Los perros fueron de diferentes razas, de siete meses a 14 años de edad y de ambos géneros (73.3% hembras y 26.7% machos). Las frecuencias de las principales alteraciones bioquímicas en suero fueron: hiperglucemia (100.0%), incremento en las concentraciones de urea (46%) y de creatinina (13%), incremento de la actividad de las enzimas ALT (50%), AST (46%), fosfatasa alcalina (56%), amilasa (20%), creatina cinasa (66%), hipercolestrolemia (66%), hiperglobulinemia (33%), hiperfosforemia (33%), hipercaliemia (43%), hiponatremia (16%), hipocloremia (46%), hipobicarbonatemia (50%), aumento de ácidos no volátiles (53%), incremento en la diferencia de iones fuertes (30%), hiperosmolalidad (23%) e hipertrigliceridemia (50%). La densidad urinaria osciló entre 1.015 y 1.064, la glucosuria presentó, en promedio, 49 mmol/L. En 10% de los casos se observó cetonuria. Las alteraciones descritas en la bioquímica clínica son importantes para el diagnóstico, manejo clínico y pronóstico de los perros con diabetes mellitus. Abstract in english The objective of this study was to describe alterations in blood clinical biochemistry and urinalysis which enable to establish the diagnosis of diabetes mellitus in dogs. Thirty cases of diabetic dogs were analyzed. The inclusion criteria were the following: hyperglycemia above 14 mmol/L, glucosuri [...] a, urine density above 1.014 and polyuria-polydipsia. Dogs of different breeds, from 7 months to 14 years of age and both genders (73.3% females and 26.7% males) were studied. The most frequent biochemical alterations in blood serum were: hyperglycemia (100%), increased urea (46%) and creatinine (13%), increase of enzyme activities ALT (50%), AST (46%), alkaline phosphatase (56%), amylase (20%), creatine kinase (66%); hypercholesterolemia (66%), hyperglobulinemia (33%), hyperphosphoremia (33%), hyperkaliemia (43%), hyponatremia (16%), hypochloremia (46%), hypobicarbonatemia (50%), increased anion gap (53%), increased strong ion difference (30%), increased serum osmolality (23%) and hypertriglyceridemia (50%). The urine density was between 1.015 and 1.064, mean value of urine glucose 49 mmol/L. Urine ketone bodies were detected in 10% of all cases. Described alterations in clinical biochemistry are important for the diagnosis, clinical care, and prognosis of dogs with diabetes mellitus.

  1. Receptor-like protein-tyrosine phosphatase alpha specifically inhibits insulin-increased prolactin gene expression

    DEFF Research Database (Denmark)

    Jacob, K K; Sap, J; Stanley, F M

    1998-01-01

    A physiologically relevant response to insulin, stimulation of prolactin promoter activity in GH4 pituitary cells, was used as an assay to study the specificity of protein-tyrosine phosphatase function. Receptor-like protein-tyrosine phosphatase alpha (RPTPalpha) blocks the effect of insulin to increase prolactin gene expression but potentiates the effects of epidermal growth factor and cAMP on prolactin promoter activity. RPTPalpha was the only protein-tyrosine phosphatase tested that did this....

  2. Colorimetric Immuno-Protein Phosphatase Inhibition Assay for Specific Detection of Microcystins and Nodularins of Cyanobacteria

    OpenAIRE

    James S. Metcalf; Bell, Steven G.; Geoffrey A. Codd

    2001-01-01

    A novel immunoassay was developed for specific detection of cyanobacterial cyclic peptide hepatotoxins which inhibit protein phosphatases. Immunoassay methods currently used for microcystin and nodularin detection and analysis do not provide information on the toxicity of microcystin and/or nodularin variants. Furthermore, protein phosphatase inhibition-based assays for these toxins are not specific and respond to other environmental protein phosphatase inhibitors, such as okadaic acid, calyc...

  3. Okadaic acid and microcystin insensitive PPP-family phosphatases may represent novel biotechnology targets

    OpenAIRE

    Uhrig, R Glen; Moorhead, Greg B.

    2011-01-01

    Reversible protein phosphorylation is of central importance to the proper cellular functioning of all living organisms. Catalyzed by the opposing reactions of protein kinases and phosphatases, dysfunction in reversible protein phosphorylation can result in a wide variety of cellular aberrations. In eukaryotic organisms there exists four classes of protein phosphatases, of which the PPP-family protein phosphatases have documented susceptibility to a range of protein and small molecule inhibito...

  4. Phosphorylation of Myosin-Binding Subunit (Mbs) of Myosin Phosphatase by Rho-Kinase in Vivo

    OpenAIRE

    Kawano, Yoji; Fukata, Yuko; Oshiro, Noriko; Amano, Mutsuki; Nakamura, Toshikazu; Ito, Masaaki; Matsumura, Fumio; Inagaki, Masaki; Kaibuchi, Kozo

    1999-01-01

    Rho-associated kinase (Rho-kinase), which is activated by the small GTPase Rho, phosphorylates myosin-binding subunit (MBS) of myosin phosphatase and thereby inactivates the phosphatase activity in vitro. Rho-kinase is thought to regulate the phosphorylation state of the substrates including myosin light chain (MLC), ERM (ezrin/radixin/moesin) family proteins and adducin by their direct phosphorylation and by the inactivation of myosin phosphatase. Here we identified the sites of phosphorylat...

  5. Protein and phosphatases pattern in the irradiated and non-irradiated pupae of med fly Ceratitis Capitata (Wield.) using different chemicals with the larval rearing media

    International Nuclear Information System (INIS)

    Thiamine, tyrosine, tryptophane and uracil were added with specific concentrations (0.5 g/kg media) to the larval rearing media of Ceratitis capitata (Wied.) as a trial to improve the quality of the produced adults to be used in sterile insect technique (SIT). The competitiveness value (C.V) of the treated and irradiated adults were so high (> 1) as compared with that of control, which is considered as a monitor for the success of SIT. The epigenetic level of protein pattern recorded a deep different pattern in each of irradiated and treated pupae and the similarity index was lowered to ? 0.08 between the control and the treated samples. Acid and alkaline phosphatases of the treated and irradiated samples were completely differed from control, which reflected a great positive variance for the use of SIT

  6. Effect of Peppermint Oil on Serum Lipid Peroxidation and Hepatic Enzymes after Immobility Stress in Mice

    OpenAIRE

    Marjani, Abdoljalal; Rahmati, Reza; Mansourian, Azad Reza; Veghary, Gholamreza

    2012-01-01

    This study was undertaken to determine the influences of various doses of peppermint oil on the hepatic en-zymes, alanine transaminase, apartate tranaminase, alkaline phosphotase and gamma glutamyl transferase and the level of malondialdehyde in the serum of mice with and without immobility stress. The mice exposed to drink water, 0.9, 27 and 60 mg/kg peppermint oil from the days 1 to 5 for a period of 4 h before and after immobility stress. Serum MDA in-creased in treatment group II, III and...

  7. Development of low cost alkaline fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Tomantschger, K.; McClusky, F.; Oporto L.; Reid, A.; Kordesch, K.

    1986-11-01

    Fuel cells as direct energy converters will find wide application in a future hydrogen economy. At the Institute for Hydrogen Systems (IHS) emphasis was placed on designing a mass producible, low, cost, alkaline, bipolar fuel cell. Carbon-filled plastics are used in the construction of the fuel cell stack. Teflon bonded, multilayer carbon electrodes have been developed. The pretreatment of carbon materials proved necessary to prolong the life of the electrodes. Electrocatalysis work resulted in the replacement of the noble metal electrocatalyst of the cathode and a significant reduction in the loading of the anode. The material cost of the alkaline, bipolar hydrogen-air fuel cell currently stands at Can Dollar 250 (US Dollar 175) per kW.

  8. Study of niobium corrosion in alkaline medium

    International Nuclear Information System (INIS)

    A comparative study of niobium electrochemical behaviour in NaOH and KOH solution, with concentrations between 0,5 and 6,1M is presented. The studies were done through electrochemicals assays, consisting in the corrosion potential and anodic and cathodic polarization curves, complemented by loss of mass experiments. The niobium anodic behaviour in alkaline medium is characterized by passivation occurrence, with a stable film formation. The Na oH solution in alkaline medium are more corrosible to niobium than the KOH solution. The loss of mass assays showed that the corrosion velocit is more dependente of hydroxide concentration in KOH medium than the NaOH medium. (C.G.C.)

  9. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    Energy Technology Data Exchange (ETDEWEB)

    Robert J. Martinez; Melanie J. Beazley; Samuel M. Webb; Martial Taillefert (co-PI); and Patricia A. Sobecky

    2007-04-19

    The overall objective of this project is to examine the activity of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium [U(VI)] phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO4 3- as a means to detoxify radionuclides and heavy metals. An experimental approach was designed to determine the extent of phosphatase activity in bacteria previously isolated from contaminated subsurface soils collected at the ERSP Field Research Center (FRC) in Oak Ridge, TN. Screening of 135 metal resistant isolates for phosphatase activity indicated the majority (75 of 135) exhibited a phosphatase-positive phenotype. During this phase of the project, a PCR based approach has also been designed to assay FRC isolates for the presence of one or more classes of the characterized non-specific acid phophastase (NSAP) genes likely to be involved in promoting U(VI) precipitation. Testing of a subset of Pb resistant (Pbr) Arthrobacter, Bacillus and Rahnella strains indicated 4 of the 9 Pbr isolates exhibited phosphatase phenotypes suggestive of the ability to bioprecipitate U(VI). Two FRC strains, a Rahnella sp. strain Y9602 and a Bacillus sp. strain Y9-2, were further characterized. The Rahnella sp. exhibited enhanced phosphatase activity relative to the Bacillus sp. Whole-cell enzyme assays identified a pH optimum of 5.5, and inorganic phosphate accumulated in pH 5.5 synthetic groundwater (designed to mimic FRC conditions) incubations of both strains in the presence of a model organophosphorus substrate provided as the sole C and P source. Kinetic experiments showed that these two organisms can grow in the presence of 200 ?M dissolved uranium and that Rahnella is much more efficient in precipitating U(VI) than Bacillus sp. The precipitation of U(VI) must be mediated by biological activity as less than 3% soluble U(VI) was removed either from the abiotic or the heat-killed cell controls. Interestingly, the pH has a strong effect on growth and U(VI) biomineralization rates by Rahnella. Thermodynamic modeling identifies autunite-type minerals [Ca(UO2)2(PO4)2] as the precipitate likely formed in the synthetic FRC groundwater conditions at all pH investigated. Extended X-ray absorption fine structure measurements have recently confirmed that the precipitate found in these incubations is an autunite and meta-autunite-type mineral. A kinetic model of U biomineralization at the different pH indicates that hydrolysis of organophosphate can be described using simple Monod kinetics and that uranium precipitation is accelerated when monohydrogen phosphate is the main orthophosphate species in solution. Overall, these experiments and ongoing soil slurry incubations demonstrate that the biomineralization of U(VI) through the activity of phosphatase enzymes can be expressed in a wide range of geochemical conditions pertaining to the FRC site.

  10. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    International Nuclear Information System (INIS)

    The overall objective of this project is to examine the activity of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium U(VI) phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO43- as a means to detoxify radionuclides and heavy metals. An experimental approach was designed to determine the extent of phosphatase activity in bacteria previously isolated from contaminated subsurface soils collected at the ERSP Field Research Center (FRC) in Oak Ridge, TN. Screening of 135 metal resistant isolates for phosphatase activity indicated the majority (75 of 135) exhibited a phosphatase-positive phenotype. During this phase of the project, a PCR based approach has also been designed to assay FRC isolates for the presence of one or more classes of the characterized non-specific acid phophastase (NSAP) genes likely to be involved in promoting U(VI) precipitation. Testing of a subset of Pb resistant (Pbr) Arthrobacter, Bacillus and Rahnella strains indicated 4 of the 9 Pbr isolates exhibited phosphatase phenotypes suggestive of the ability to bioprecipitate U(VI). Two FRC strains, a Rahnella sp. strain Y9602 and a Bacillus sp. strain Y9-2, were further characterized. The Rahnella sp. exhibited enhanced phosphatase activity relative to the Bacillus sp. Whole-cell enzyme assays identified a pH optimum of 5.5, and inorganic phosphate accumulated in pH 5.5 synthetic groundwater (designed to mimic FRC conditions) incubations of both strains in the presence of a model organophosphorus substrate provided as the sole C and P source. Kinetic experiments showed that these two organisms can grow in the presence of 200 (micro)M dissolved uranium and that Rahnella is much more efficient in precipitating U(VI) than Bacillus sp. The precipitation of U(VI) must be mediated by biological activity as less than 3% soluble U(VI) was removed either from the abiotic or the heat-killed cell controls. Interestingly, the pH has a strong effect on growth and U(VI) biomineralization rates by Rahnella. Thermodynamic modeling identifies autunite-type minerals Ca(UO2)2(PO4)2 as the precipitate likely formed in the synthetic FRC groundwater conditions at all pH investigated. Extended X-ray absorption fine structure measurements have recently confirmed that the precipitate found in these incubations is an autunite and meta-autunite-type mineral. A kinetic model of U biomineralization at the different pH indicates that hydrolysis of organophosphate can be described using simple Monod kinetics and that uranium precipitation is accelerated when monohydrogen phosphate is the main orthophosphate species in solution. Overall, these experiments and ongoing soil slurry incubations demonstrate that the biomineralization of U(VI) through the activity of phosphatase enzymes can be expressed in a wide range of geochemical conditions pertaining to the FRC site

  11. Osteocalcin as an indicator of bone metabolism during spaceflight

    Science.gov (United States)

    Patterson-Buckendahl, P. E.; Cann, C. E.; Grindeland, R. E.; Arnaud, S. B.; Martin, R. B.

    1985-01-01

    The effect of microgravity on osteocalcin (OC) is investigated in rats flown on Spacelab 3. Serum, Ca, Pi, total protein, alkaline phosphatase, and OC contents, and the breaking strength of the humerus of control and Spacelab rats are calculated; the procedures utilized for these analyses are described. It is detected that the OC is reduced, and the serum and alkaline phosphatase are unaffected by microgravity.

  12. Atypical form of active melorheostosis and its treatment with bisphosphonate

    Energy Technology Data Exchange (ETDEWEB)

    Donath, Judit; Poor, Gyula; Kiss, Csaba [National Institute of Rheumatology and Physiotherapy, 1027 Budapest (Hungary); Fornet, Bela [Semmelweis University Department of Radiology, Budapest (Hungary); Genant, Harry [University of California Department of Radiology, San Francisco, California (United States)

    2002-12-01

    We present the case of a 38-year-old man in whom extensive bilateral melorheostosis was associated with elevated serum alkaline phosphatase, swelling of the right foot and progressive deformity of the left hand, left leg and right foot. Radiography, computed tomography and bone scintigraphy were performed. Following treatment with bisphosphonate (30 mg/day of pamidronate for 6 days) infusion, the pain and swelling of his right foot showed improvement and his elevated serum alkaline phosphatase decreased. (orig.)

  13. Atypical form of active melorheostosis and its treatment with bisphosphonate

    International Nuclear Information System (INIS)

    We present the case of a 38-year-old man in whom extensive bilateral melorheostosis was associated with elevated serum alkaline phosphatase, swelling of the right foot and progressive deformity of the left hand, left leg and right foot. Radiography, computed tomography and bone scintigraphy were performed. Following treatment with bisphosphonate (30 mg/day of pamidronate for 6 days) infusion, the pain and swelling of his right foot showed improvement and his elevated serum alkaline phosphatase decreased. (orig.)

  14. CARINA alkalinity data in the Atlantic Ocean

    Directory of Open Access Journals (Sweden)

    A. Velo

    2009-08-01

    Full Text Available Data on carbon and carbon-relevant hydrographic and hydrochemical parameters from previously non-publicly available cruise data sets in the Arctic, Atlantic and Southern Ocean have been retrieved and merged to a new database: CARINA (CARbon IN the Atlantic.

    These data have gone through rigorous quality control (QC procedures to assure the highest possible quality and consistency. The data for most of the measured parameters in the CARINA data base were objectively examined in order to quantify systematic differences in the reported values, i.e. secondary quality control. Systematic biases found in the data have been corrected in the data products, i.e. three merged data files with measured, calculated and interpolated data for each of the three CARINA regions; Arctic, Atlantic and Southern Ocean. Out of a total of 188 cruise entries in the CARINA database, 98 were conducted in the Atlantic Ocean and of these, 75 cruises report alkalinity values.

    Here we present details of the secondary QC on alkalinity for the Atlantic Ocean part of CARINA. Procedures of quality control, including crossover analysis between cruises and inversion analysis of all crossover data are briefly described. Adjustments were applied to the alkalinity values for 16 of the cruises in the Atlantic Ocean region. With these adjustments the CARINA database is consistent both internally as well as with GLODAP data, an oceanographic data set based on the World Hydrographic Program in the 1990s. Based on our analysis we estimate the internal accuracy of the CARINA-ATL alkalinity data to be 3.3 ?mol kg?1. The CARINA data are now suitable for accurate assessments of, for example, oceanic carbon inventories and uptake rates and for model validation.

  15. Alkaline decomposition of synthetic jarosite with arsenic

    OpenAIRE

    Patiño, Francisco; Flores, Mizraim U; Reyes, Iván A; Reyes, Martín; Hernández, Juan; Rivera, Isauro; Juárez, Julio C

    2013-01-01

    The widespread use of jarosite-type compounds to eliminate impurities in the hydrometallurgical industry is due to their capability to incorporate several elements into their structures. Some of these elements are of environmental importance (Pb2+, Cr6+, As5+, Cd2+, Hg2+). For the present paper, AsO43- was incorporated into the lattice of synthetic jarosite in order to carry out a reactivity study. Alkaline decomposition is characterized by removal of sulfate and potassium ions from the latti...

  16. 2nd Generation Alkaline Electrolysis : Final report

    DEFF Research Database (Denmark)

    Yde, Lars; Kjartansdóttir, Cecilia Kristin

    2013-01-01

    This report provides the results of the 2nd Generation Alkaline Electrolysis project which was initiated in 2008. The project has been conducted from 2009-2012 by a consortium comprising Århus University Business and Social Science – Centre for Energy Technologies (CET (former HIRC)), Technical University of Denmark – Mechanical Engineering (DTU-ME), Technical University of Denmark – Energy Conversion (DTU-EC), FORCE Technology and GreenHydrogen.dk. The project has been supported by EUDP.

  17. Bifunctional Catalysts for Alkaline Fuel Cells.

    Czech Academy of Sciences Publication Activity Database

    Klápšt?, B?etislav; Vondrák, Ji?í; Velická, Jana

    Vol. 1. Brno : Akademické nakladatelství CERM, 2000 - (Vondrák, J.; Sedla?íková, M.), s. 24.1-24.4 ISBN 80-214-1614-9. [Advanced Batteries and Accumulators /1./. Brno (CZ), 28.08.2000-01.09.2000] R&D Projects: GA MŠk ME 216 Institutional research plan: CEZ:AV0Z4032918; CEZ:MSM 262200010 Keywords : alkaline * catalysts * electrochemistry Subject RIV: CG - Electrochemistry

  18. Regulation of a Novel Phosphatase Common in Marine Bacteria

    Science.gov (United States)

    D'Ambrosio, L. A.; Sebastian, M.; Ammerman, J. W.

    2008-12-01

    The assessment of the in situ phosphorus (P) status of marine bacteria will help us understand the P cycle in the ocean and its effect on bacterial ecology and diversity. A possible marker of P status is phoX, a recently discovered, widespread phosphatase gene in marine bacteria. This gene has been shown to encode for the major P-regulated phosphatase in Silicibacter pomeroyi. However, further information about the regulation of this gene is required in order to use it as a molecular marker of P status. The aim of this study was to elucidate whether it PhoX is solely responding to P starvation or it is induced upon other conditions. We investigated the effect of carbon and iron limitation and the addition of organic P on phoX expression and activity. S. pomeroyi was grown in minimal media in the presence and absence of phosphate, glucose, and glucose-6-phosphate (G6P), and under low and high iron conditions. Growth and phosphatase activity were monitored and phoX gene expression was determined by RT-PCR techniques. phoX expression was induced in all the treatments that lacked phosphate, but not by the presence of G6P, indicating that PhoX is not regulated by organic P substrates. The effect of carbon limitation on phoX expression could not be assessed, since there was no growth in any of the treatments that lacked glucose. Iron limitation in the presence of phosphate had no effect on PhoX activity, but the effect on phoX gene expression is still unclear and requires further study.

  19. Characterization of the protein tyrosine phosphatase PRL from Entamoeba histolytica.

    Science.gov (United States)

    Ramírez-Tapia, Ana Lilia; Baylón-Pacheco, Lidia; Espíritu-Gordillo, Patricia; Rosales-Encina, José Luis

    2015-12-01

    Protein tyrosine phosphatase of regenerating liver (PRL) is a group of phosphatases that has not been broadly studied in protozoan parasites. In humans, PRLs are involved in metastatic cancer, the promotion of cell migration and invasion. PTPs have been increasingly recognized as important effectors of host-pathogen interactions. We characterized the only putative protein tyrosine phosphatase PRL (PTP EhPRL) in the eukaryotic human intestinal parasite Entamoeba histolytica. Here, we reported that the EhPRL protein possessed the classical HCX5R catalytic motif of PTPs and the CAAX box characteristic of the PRL family and exhibited 31-32% homology with the three human PRL isoforms. In amebae, the protein was expressed at low but detectable levels. The recombinant protein (rEhPRL) had enzymatic activity with the 3-o-methyl fluorescein phosphate (OMFP) substrate; this enzymatic activity was inhibited by the PTP inhibitor o-vanadate. Using immunofluorescence we showed that native EhPRL was localized to the cytoplasm and plasma membrane. When the trophozoites interacted with collagen, EhPRL relocalized over time to vesicle-like structures. Interaction with fibronectin increased the presence of the enzyme in the cytoplasm. Using RT-PCR, we demonstrated that EhPRL mRNA expression was upregulated when the trophozoites interacted with collagen but not with fibronectin. Trophozoites recovered from amoebic liver abscesses showed higher EhPRL mRNA expression levels than normal trophozoites. These results strongly suggest that EhPRL may play an important role in the biology and adaptive response of the parasite to the host environment during amoebic liver abscess development, thereby participating in the pathogenic mechanism. PMID:26431820

  20. Degradation of polymers in an alkaline environment

    Energy Technology Data Exchange (ETDEWEB)

    Borges, P. G.; Thorpe, S. J.; Venter, R. D. [Toronto Univ., Toronto, ON (Canada)

    1999-10-01

    To introduce widespread use of hydrogen as a source of fuel, it is essential to significantly reduce the cost of electrolysers and fuel cells, preferably by substituting degradation-resistant low cost alternatives for traditional materials. An experimental apparatus for evaluating short-term degradation characteristics of polymers in 25 per cent potassium hydroxide aqueous solution at 70 and at 85 degrees C was constructed to determine changes in physical appearance, weight, volume and dimensions as a function of immersion time from 0 to 670 hours. Polymers that were part of this test included chlorinated polyvinyl chloride (CPVC), polypropylene (PP),and a polyurethane/polyester blend. Polypropylene demonstrated the greatest degradation resistance to the alkaline environment at both temperatures. CPVC discolored and the polyurethane/polyester blend deteriorated into smaller pieces when subjected to identical conditions. Resistance to degradation in an alkaline environment is a primary requirement for the selection of polymers for use as structural materials in alkaline water analyzers, which in turn is fundamental to the widespread adoption of a hydrogen economy. 6 refs., 13 figs.

  1. Heavy water production by alkaline water electrolysis

    International Nuclear Information System (INIS)

    Several heavy water isotope production processes are reported in literature. Water electrolysis in combination with catalytic exchange CECE process is considered as a futuristic process to increase the throughput and reduce the cryogenic distillation load but the application is limited due to the high cost of electricity. Any improvement in the efficiency of electrolyzers would make this process more attractive. The efficiency of alkaline water electrolysis is governed by various phenomena such as activation polarization, ohmic polarization and concentration polarization in the cell. A systematic study on the effect of these factors can lead to methods for improving the efficiency of the electrolyzer. A bipolar and compact type arrangement of the alkaline water electrolyzer leads to increased efficiency and reduced inventory in comparison to uni-polar tank type electrolyzers. The bipolar type arrangement is formed when a number of single cells are stacked together. Although a few experimental studies have been reported in the open literature, CFD simulation of a bipolar compact alkaline water electrolyzer with porous electrodes is not readily available.The principal aim of this study is to simulate the characteristics of a single cell compact electrolyzer unit. The simulation can be used to predict the Voltage-Current Density (V-I) characteristics, which is a measure of the efficiency of the process.The model equations were solved using COMSOL multi-physics software. The simulated V-I characteristic is compared with the experimental data

  2. Promoting Uranium Immobilization by the Activities of Microbial Phosphatases

    Energy Technology Data Exchange (ETDEWEB)

    Martinez, Robert J.; Beazley, Melanie J.; Wilson, Jarad J.; Taillefert, Martial; Sobecky, Patricia A.

    2005-04-05

    The overall goal of this project is to examine the role of nonspecific phosphohydrolases present in naturally occurring subsurface microorganisms for the purpose of promoting the immobilization of radionuclides through the production of uranium [U(VI)] phosphate precipitates. Specifically, we hypothesize that the precipitation of U(VI) phosphate minerals may be promoted through the microbial release and/or accumulation of PO{sub 4}{sup 3-}. During this phase of the project we have been conducting assays to determine the effects of pH, inorganic anions and organic ligands on U(VI) mineral formation and precipitation when FRC bacterial isolates were grown in simulated groundwater medium. The molecular characterization of FRC isolates has also been undertaken during this phase of the project. Analysis of a subset of gram-positive FRC isolates cultured from FRC soils (Areas 1, 2 and 3) and background sediments have indicated a higher percentage of isolates exhibiting phosphatase phenotypes (i.e., in particular those surmised to be PO{sub 4}{sup 3-}-irrepressible) relative to isolates from the reference site. A high percentage of strains that exhibited such putatively PO{sub 4}{sup 3-}-irrepressible phosphatase phenotypes were also resistant to the heavy metals lead and cadmium. Previous work on FRC strains, including Arthrobacter, Bacillus and Rahnella spp., has demonstrated differences in tolerance to U(VI) toxicity (200 {micro}M) in the absence of organophosphate substrates. For example, Arthrobacter spp. exhibited the greatest tolerance to U(VI) while the Rahnella spp. have been shown to facilitate the precipitation of U(VI) from solution and the Bacillus spp. demonstrate the greatest sensitivity to acidic conditions and high concentrations of U(VI). PCR-based detection of FRC strains are being conducted to determine if non-specific acid phosphatases of the known molecular classes [i.e., classes A, B and C] are present in these FRC isolates. Additionally, these amplified phosphatases are being analyzed to determine whether or not there is evidence for the horizontal transfer of such genes amongst subsurface microbial populations. Microbially precipitated U(VI) phosphate minerals will be further analyzed via capillary electrophoresis and extended x-ray absorption fine structure spectroscopy to determine uranium speciation.

  3. Tea Contains Potent Inhibitors of Tyrosine Phosphatase PTP1B

    OpenAIRE

    Ma, Junfeng; LI Zhe; Xing, Shu; Ho, Wanting Tina; Fu, Xueqi; Zhao, Zhizhuang Joe

    2011-01-01

    Tea is widely consumed all over the world. Studies have demonstrated the role of tea in prevention and treatment of various chronic diseases including diabetes and obesity, but the underlying mechanism is unclear. PTP1B is a widely expressed tyrosine phosphatase which has been defined as a target for therapeutic drug development to treat diabetes and obesity. In screening for inhibitors of PTP1B, we found that aqueous extracts of teas exhibited potent PTP1B inhibitory effects with an IC50 val...

  4. PROTEN TYROSINE PHOSPHATASE ACTIVITY IN RAT ASCITES HEPATOMA CELLS

    Directory of Open Access Journals (Sweden)

    M.Saadat

    1998-10-01

    Full Text Available Protein tyrosine phosphatases (PTPases regulate tyrosine phosphorylation of target proteins involved in several aspects of cellular functions. Enzyme activities of the PTPases in cytosolic and particulate fractions of rat ascites hepatoma cell lines were determined and compared with those of normal rat liver. Our present data revealed that although there was no neoplatic-specific alteration of the PTPase activity in examined hepatomas, the activity in particulate fractions of island type of hepatomas was remarkably decreased compared with either rat liver or free type hepatomas.

  5. The fate of added alkalinity in model scenarios of ocean alkalinization

    Science.gov (United States)

    Ferrer González, Miriam; Ilyina, Tatiana

    2014-05-01

    The deliberate large-scale manipulation of the Earth's climate (geo-engineering) has been proposed to mitigate climate change and ocean acidification. Whilst the mitigation potential of these technologies could sound promising, they may also pose many environmental risks. Our research aims at exploring the ocean-based carbon dioxide removal method of alkalinity enhancement. Its mitigation potential to reduce atmospheric CO2 and counteract the consequences of ocean acidification, risks and unintended consequences are studied. In order to tackle these questions, different scenarios are implemented in the state-of-the-art Earth system model of the Max Planck Institute for Meteorology. The model configuration is based on the 5th phase of the coupled model intercomparison project following a high CO2 future climate change scenario RCP8.5 (in which radiative forcing rises to 8.5 W/m² in 2100). Two different scenarios are performed where the alkalinity is artificially added globally uniformly in the upper ocean. In the first scenario, alkalinity is increased as a pulse by doubling natural values of the first 12 meters. In the second scenario we add alkalinity into the same ocean layer such that the atmospheric CO2 concentration is reduced from RCP8.5 to RCP4.5 levels (with the radiative forcing of 4.5 W/m² in 2100). We investigate the fate of the added alkalinity in these two scenarios and compare the differences in alkalinity budgets. In order to increase oceanic CO2 uptake from the atmosphere, enhanced alkalinity has to stay in the upper ocean. Once the alkalinity is added, it will become part of the biogeochemical cycles and it will be distributed with the ocean currents. Therefore, we are particularly interested in the residence time of the added alkalinity at the surface. Variations in CO2 partial pressure, seawater pH and saturation state of carbonate minerals produced in the implemented scenarios will be presented. Collateral changes in ocean biogeochemistry and climate will be also discussed.

  6. 18O isotope effect in 13C nuclear magnetic resonance spectroscopy. Part 9. Hydrolysis of benzyl phosphate by phosphatase enzymes and in acidic aqueous solutions

    International Nuclear Information System (INIS)

    The 18O isotope-induced shifts in 13C and 31P nuclear magnetic resonance (NMR) spectroscopy were used to establish the position of bond cleavage in the phosphatase-catalyzed and acid-catalyzed hydrolysis reactions of benzyl phosphate. The application of the 18O-isotope effect in NMR spectroscopy affords a continuous, nondestructive assay method for following the kinetics and position of bond cleavage in the hydrolytic process. The technique provides advantages over most discontinuous methods in which the reaction components must be isolated and converted to volatile derivatives prior to analysis. In the present study, [?-13C,ester-18O]benzyl phosphate and [ester-18O]benzyl phosphate were synthesized for use in enzymatic and nonenzymatic studies. Hydrolysis reactions catalyzed by the alkaline phosphatase from E. coli and by the acid phosphatases isolated from human prostate and human liver were all accompanied by cleavage of the substrate phosphorus-oxygen bond consistent with previously postulated mechanisms involving covalent phosphoenzyme intermediates. An extensive study of the acid-catalyzed hydrolysis of benzyl phosphate at 750C revealed that the site of bond cleavage is dependent on pH. At pH less than or equal to 1.3, the hydrolysis proceeds with C-O bond cleavage; at 1.3 2H]Benzyl phosphate was also synthesized. Hydrolysis of this chiral benzyl derivative demonstrated that the acid-catalyzed C-O bond scission of benzyl phosphate proceeds by an A-1 (S/sub N/1) mechanism with 70% racemization and 30% inversion at carbon. 37 references, 4 figures, 2 tables

  7. Identification of a non-purple tartrate-resistant acid phosphatase: an evolutionary link to Ser/Thr protein phosphatases?

    Directory of Open Access Journals (Sweden)

    Hume David A

    2008-09-01

    Full Text Available Abstract Background Tartrate-resistant acid phosphatases (TRAcPs, also known as purple acid phosphatases (PAPs, are a family of binuclear metallohydrolases that have been identified in plants, animals and fungi. The human enzyme is a major histochemical marker for the diagnosis of bone-related diseases. TRAcPs can occur as a small form possessing only the ~35 kDa catalytic domain, or a larger ~55 kDa form possessing both a catalytic domain and an additional N-terminal domain of unknown function. Due to its role in bone resorption the 35 kDa TRAcP has become a promising target for the development of anti-osteoporotic chemotherapeutics. Findings A new human gene product encoding a metallohydrolase distantly related to the ~55 kDa plant TRAcP was identified and characterised. The gene product is found in a number of animal species, and is present in all tissues sampled by the RIKEN mouse transcriptome project. Construction of a homology model illustrated that six of the seven metal-coordinating ligands in the active site are identical to that observed in the TRAcP family. However, the tyrosine ligand associated with the charge transfer transition and purple color of TRAcPs is replaced by a histidine. Conlusion The gene product identified here may represent an evolutionary link between TRAcPs and Ser/Thr protein phosphatases. Its biological function is currently unknown but is unlikely to be associated with bone metabolism.

  8. Molecular cloning, expression, and characterization of PTPA, a protein that activates the tyrosyl phosphatase activity of protein phosphatase 2A.

    Science.gov (United States)

    Cayla, X; Van Hoof, C; Bosch, M; Waelkens, E; Vandekerckhove, J; Peeters, B; Merlevede, W; Goris, J

    1994-06-01

    PTPA, or phosphotyrosyl phosphatase activator, is a protein that stimulates the tyrosyl phosphatase activity of protein phosphatase 2A in an ATP, Mg(2+)-requiring reaction (Cayla, X., Goris, J., Hermann, J., Hendrix, P., Ozon, R., and Merevede, W. (1990) Biochemistry 29, 658-667). We constructed oligonucleotide probes based on the amino acid sequences of peptides isolated from purified PTPA and used them to probe rabbit muscle and human heart cDNA libraries. A putative full-length clone was isolated from the rabbit skeletal muscle as well as from the human heart library. The nucleotide sequence of both clones contains an open reading frame of 969 nucleotides starting from an assigned initial ATG codon and encodes for a protein of 323 amino acids. The predicted rabbit and human PTPA protein sequences show an identity of 96.6%. The predicted protein matched all the peptide sequences obtained from the rabbit skeletal muscle protein. Bacterially expressed protein, as well as the in vitro reticulocyte lysate translation product, comigrated with the purified 37-kDa protein on sodium dodecyl sulfate-polyacrylamide gels. Both proteins reacted with immunopurified, anti-PTPA polyclonal antiserum. The recombinant protein was a soluble and active protein. Northern blot analysis revealed two transcripts of 2.8 and 4 kilobases, respectively, in human placenta but only one 2.8-kilobase transcript in rabbit and rat tissues. High levels of PTPA mRNA were detected in testis, which contrasted with the low levels present in skeletal muscle. PMID:8195217

  9. The TriTryp Phosphatome: analysis of the protein phosphatase catalytic domains

    Directory of Open Access Journals (Sweden)

    Huxley-Jones Julie

    2007-11-01

    Full Text Available Abstract Background The genomes of the three parasitic protozoa Trypanosoma cruzi, Trypanosoma brucei and Leishmania major are the main subject of this study. These parasites are responsible for devastating human diseases known as Chagas disease, African sleeping sickness and cutaneous Leishmaniasis, respectively, that affect millions of people in the developing world. The prevalence of these neglected diseases results from a combination of poverty, inadequate prevention and difficult treatment. Protein phosphorylation is an important mechanism of controlling the development of these kinetoplastids. With the aim to further our knowledge of the biology of these organisms we present a characterisation of the phosphatase complement (phosphatome of the three parasites. Results An ontology-based scan of the three genomes was used to identify 86 phosphatase catalytic domains in T. cruzi, 78 in T. brucei, and 88 in L. major. We found interesting differences with other eukaryotic genomes, such as the low proportion of tyrosine phosphatases and the expansion of the serine/threonine phosphatase family. Additionally, a large number of atypical protein phosphatases were identified in these species, representing more than one third of the total phosphatase complement. Most of the atypical phosphatases belong to the dual-specificity phosphatase (DSP family and show considerable divergence from classic DSPs in both the domain organisation and sequence features. Conclusion The analysis of the phosphatome of the three kinetoplastids indicates that they possess orthologues to many of the phosphatases reported in other eukaryotes, including humans. However, novel domain architectures and unusual combinations of accessory domains, suggest distinct functional roles for several of the kinetoplastid phosphatases, which await further experimental exploration. These distinct traits may be exploited in the selection of suitable new targets for drug development to prevent transmission and spread of the diseases, taking advantage of the already extensive knowledge on protein phosphatase inhibitors.

  10. Perspective: Tyrosine phosphatases as novel targets for antiplatelet therapy.

    Science.gov (United States)

    Tautz, Lutz; Senis, Yotis A; Oury, Cécile; Rahmouni, Souad

    2015-06-15

    Arterial thrombosis is the primary cause of most cases of myocardial infarction and stroke, the leading causes of death in the developed world. Platelets, highly specialized cells of the circulatory system, are key contributors to thrombotic events. Antiplatelet drugs, which prevent platelets from aggregating, have been very effective in reducing the mortality and morbidity of these conditions. However, approved antiplatelet therapies have adverse side effects, most notably the increased risk of bleeding. Moreover, there remains a considerable incidence of arterial thrombosis in a subset of patients receiving currently available drugs. Thus, there is a pressing medical need for novel antiplatelet agents with a more favorable safety profile and less patient resistance. The discovery of novel antiplatelet targets is the matter of intense ongoing research. Recent findings demonstrate the potential of targeting key signaling molecules, including kinases and phosphatases, to prevent platelet activation and aggregation. Here, we offer perspectives to targeting members of the protein tyrosine phosphatase (PTP) superfamily, a major class of enzymes in signal transduction. We give an overview of previously identified PTPs in platelet signaling, and discuss their potential as antiplatelet drug targets. We also introduce VHR (DUSP3), a PTP that we recently identified as a major player in platelet biology and thrombosis. We review our data on genetic deletion as well as pharmacological inhibition of VHR, providing proof-of-principle for a novel and potentially safer VHR-based antiplatelet therapy. PMID:25921264

  11. Spatial control of protein phosphatase 2A (de)methylation

    International Nuclear Information System (INIS)

    Reversible methylation of the protein phosphatase 2A catalytic subunit (PP2AC) is an important regulatory mechanism playing a crucial role in the selective recruitment of regulatory B subunits. Here, we investigated the subcellular localization of leucine carboxyl methyltransferase (LCMT1) and protein phosphatase methylesterase (PME-1), the two enzymes catalyzing this process. The results show that PME-1 is predominantly localized in the nucleus and harbors a functional nuclear localization signal, whereas LCMT1 is underrepresented in the nucleus and mainly localizes to the cytoplasm, Golgi region and late endosomes. Indirect immunofluorescence with methylation-sensitive anti-PP2AC antibodies revealed a good correlation with the methylation status of PP2AC, demethylated PP2AC being substantially nuclear. Throughout mitosis, demethylated PP2AC is associated with the mitotic spindle and during cytokinesis with the cleavage furrow. Overexpression of PME-1, but not of an inactive mutant, results in increased demethylation of PP2AC in the nucleus, whereas overexpression of a cytoplasmic PME-1 mutant lacking the NLS results in increased demethylation in the cytoplasm-in all cases, however, without any obvious functional consequences. PME-1 associates with an inactive PP2A population, regardless of its esterase activity or localization. We propose that stabilization of this inactive, nuclear PP2A pool is a major in vivo function of PME-1

  12. Electron emission of alkaline and alkaline-earth vapors adsorbed on refractory metals

    International Nuclear Information System (INIS)

    This work has been carried out as an aid to research on thermionic converter electrodes. The improvement of the converter efficiency makes it necessary to explain physically the adsorption process, and to determine theoretically its laws. The Gyftopoulos, Levine, Steiner's latest theory has been used to plot, by means of a computer, the 'S' shaped curves which bind the electron emission of adsorbed alkaline and alkaline-earth metals to their temperature and to the substrate temperature. 21 examples are presented, some of them being studied with crystallographic orientation. (authors)

  13. Hemograma e perfil bioquímico sérico, inclusive hemogasométrico, de bezerros infectados experimentalmente com Salmonella Dublin / Blood cell counts and serum biochemical profile, including blood gas levels, in Salmonella Dublin-infected calves

    Scientific Electronic Library Online (English)

    D.G., Silva; P.R.L., Silva; J.J., Fagliari.

    2010-04-01

    Full Text Available O objetivo do estudo foi avaliar o hemograma e o perfil bioquímico sérico, inclusive hemogasométrico, de bezerros infectados experimentalmente com Salmonella Dublin. Foram utilizados 12 bezerros sadios da raça Holandesa com 10 a 15 dias de idade, distribuídos aleatoriamente em dois grupos experiment [...] ais: grupo-controle (n= 6) e grupo infectado com 10(8)UFC de Salmonella Dublin (n=6). Os bezerros foram submetidos ao exame físico diário, e as amostras de sangue foram coletadas minutos antes da inoculação (0h) e 24, 48, 72, 96, 120, 144 e 168h após a inoculação. Além do hemograma e das análises hemogasométricas, foram mensuradas as atividades séricas das enzimas aspartato aminotransferase (AST), fosfatase alcalina (ALP), creatina cinase (CK), gamaglutamiltransferase (GGT) e lactato desidrogenase (LDH), e os teores de albumina, bilirrubinas, cálcio total, cálcio ionizado, sódio, potássio, cloretos, creatinina, ferro, fibrinogênio, fósforo, glicose, magnésio, proteína totais e ureia. As principais alterações foram: redução das concentrações de albumina, ferro, glicose, magnésio e proteína total, aumento do teor de fibrinogênio, leucocitose e acidose metabólica e hiponatremia. Abstract in english The blood cell counts and biochemical profile, including blood gas levels, were evaluated, in Salmonella Dublin-infected calves. Twelve healthy 10 to 15-day old Holstein calves were randomly allotted into two groups: control (n=6) and group orally infected with 10(8) CFU Salmonella Dublin (n=6). The [...] calves were submitted to physical examination and the blood samples were taken just before the inoculation (0h) and at 24, 48, 72, 96, 120, 144, and 168h later. Besides, blood cell counts, blood gas levels, and the serum concentrations of aspartate aminotransferase (AST), alkaline phosphatase (ALP), creatine kinase (CK), gammaglutamyltransferase (GGT), lactate desidrogenase (LDH), albumin, bilirubin, total calcium, ionic calcium, sodium, potassium, chlorides, creatinine, iron, fibrinogen, phosphorus, glucose, magnesium, total protein, and urea were measured. The main findings included decrease in albumin, iron, glucose, magnesium, and total protein concentrations, and increase in fibrinogen level, leukocytosis, metabolic acidosis, and hyponatremia.

  14. Telopéptido carboxilo terminal del colágeno tipo I (b-CTX sérico y compromiso óseo en la insuficiencia renal crónica Serum b-Type I collagen carboxyterminal telopeptide (b-CTXs and bone involvement in chronic renal failure

    Directory of Open Access Journals (Sweden)

    Beatriz Oliveri

    2005-08-01

    Full Text Available La osteodistrofia renal (ODR se caracteriza por alteraciones óseas. Se evaluaron métodos bioquímicos alternativos a la biopsia ósea en pacientes renales para determinar cambios rápidos del remodelamiento óseo en 43 pacientes predialíticos (PD y 49 hemodializados (HD. Los PD presentaron fosfatemia, fosfatasa alcalina ósea (FAO, hormona paratiroidea intacta (PTHi y b-telopéptido carboxilo terminal del colágeno tipo I (bCTXs mayores y clearence de creatinina (Ccr menores (p40 ml/min. En PD, bCTXs (pAn increase in parathyroid hormone (PTH levels in chronic renal failure (CRF induces bone abnormalities known as renal osteodystrophy (ROD. The aim of the present study was to evaluate alternative biochemical methods to bone biopsy, to evaluate changes in bone remodeling in renal patients. Intact PTH (iPTH and bone markers were measured in 43 predialysis (PD, 49 hemodialysis patients (HD and 185 controls. bCTXs, bone alkaline phosphatase (bAL, iPTH were higher and creatinine clearance (Ccr was lower in PD and HD compared with controls (p40 ml/min. bCTXs (p<0.05 in PD and bCTXs and bAL in HD patients were higher than controls, even when iPTH was within normal range (<65 pg/ml. Patients with severe secondary hyperparathyroidism showed higher bone markers than patients with normal or moderately increased iPTH (p<0.001. These results suggest that even when there is no increase in iPTH, bone remodeling increases (possibly due to other factors exhibiting higher bone resorption, and bCTXs would seem to be an adequate non-invasive tool to assess early bone changes in CRF and prevent future fractures. Bone marker measurements in ROD would be useful to identify patients who may require bone biopsy. However, further studies comparing both methods must be performed before replacing bone biopsy with serum b-CTX.

  15. Free serum thyroxine

    International Nuclear Information System (INIS)

    The use of radiommunoassay (RIA) tehcniques has increased the diagnosis of thyroid functional alterations. A solid phase RIA method for free thyroxine (FT4) measurement was tested. Serum FT4, Total T4, T3 and TSH were determined by radioimmunoassay in 179 subjects. One hundred twenty two patients were normal (8 to 75 years old); FT4 was 1.42 +- 0.03 ng/100 ml (avg. value and std. error). In 27 cases of thyrotoxicosis the values were 4.66 +- 0.48 ng/100 ml and in 15 cases of hypothyroidism 0.50 +- 0.06 ng/100 ml (statistics probability <0,01). Four euthyroid patients under estrogen treatment, 9 hypothyroid patients under l-thyroxine treatment, one hyperthyroid patient under antithyroid drugs and one hyperthyroid patient under estrogen treatment, were studied. Total T4 and T3 values were in agreement with the variations of the serum protein transport capacity, and for that reason did not show the real thyroid function state. Clinical decision making analysis (with Receiver Operating Characteristic (ROC) curves) indicate that FT4 is the best choice for hyperthyroidism diagnosis and T4 for hypothyroidism. Cost/benefit analysis indicates also that FT4 has the best ratio. (author)

  16. Preparation, Biodegradation of Coconut Oil Driven Chemically Modified Bovine Serum Albumin Microparticles of Encapsulated Cicer arietinum Amylase and Study of Their Application in Washing Detergents

    OpenAIRE

    Kirti Rani; Vartika Mehta

    2014-01-01

    In present work, Cicer arietinum amylase was encapsulated by emulsification through covalent coupling by glutaraldehyde into chemically modified bovine serum albumin. Biodegradation of coconut oil driven emulsified bovine serum albumin encapsulated Cicer arietinum amylase was carried out by the alkaline protease for its controlled and sustained release of encapsulated enzyme from prepared microparticles of encapsulated Cicer arietinum amylase and its stability increased up to 6 months as comp...

  17. Correlations of Serum IL-6 Levels and Prolidase Activity Between Bone Turnover Markers and Bone Mineral Density in Postmenopausal Women With and Without Osteoporosis

    OpenAIRE

    Toker, Aysun

    2007-01-01

    Aim: The purpose of this study was to measure serum prolidase activity and interleukin-2 and