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Alkaline Phosphatase (alkaline + phosphatase)
Kinds of Alkaline Phosphatase Terms modified by Alkaline Phosphatase Selected AbstractsTHERMAL INACTIVATION KINETICS OF ALKALINE PHOSPHATASE IN BUFFER AND MILKJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 3 2006S. FADILO ABSTRACT A detailed kinetic study on the thermal inactivation of alkaline phosphatase (ALP) added into buffer and pasteurized milk and for ALP naturally present in raw cow's milk has been performed. Kinetic parameters (rate constant, k; decimal reduction time, D; activation energy, Ea; and z value) were evaluated based on the first-order rate model at 50,80C. The temperature sensitivity of the kinetic parameters was evaluated considering the Arrhenius-type Ea model. All kinetic behaviors were well described by the first-order model (r2 > 0.91). The D values increased with increasing temperature. Higher temperatures resulted in higher rates of enzyme inactivation as indicated by lower D values and higher k values. There are significant differences (P < 0.01) among the D values for ALP in buffer and milk at treated temperatures. The rate of enzyme inactivation was much more rapid in buffer than in pasteurized milk. The evaluated Ea values for ALP added into the buffer and pasteurized milk, and for ALP naturally present in raw milk were 97.2, 149.9 and 207.8 kJ/mol, respectively. The inactivation kinetics of ALP during heat treatment was found to be dependent on the composition of the medium, and the time and temperature of the heat treatment. [source] Platform for Highly Sensitive Alkaline Phosphatase-Based Immunosensors Using 1-Naphthyl Phosphate and an Avidin-Modified Indium Tin Oxide ElectrodeELECTROANALYSIS, Issue 19 2009Abdul Aziz Abstract We report a versatile platform for highly sensitive alkaline phosphatase (ALP)-based electrochemical biosensors that uses an avidin-modified indium tin oxide (ITO) electrode as a sensing electrode and 1-naphthyl phosphate (NPP) as an ALP substrate. Almost no electrocatalytic activity of NPP and good electrocatalytic activity of 1-naphthol (ALP product) on the ITO electrodes allow a high signal-to-background ratio. The effective surface covering of avidin on the ITO electrodes allows very low levels of nonspecific binding of proteins to the sensing electrodes. The platform technology is used to detect mouse IgG with a detection limit of 1.0,pg/mL. [source] Transgenic mice expressing a dual, CRE-inducible reporter for the analysis of axon guidance and synaptogenesis,GENESIS: THE JOURNAL OF GENETICS AND DEVELOPMENT, Issue 6 2007Aurora Badaloni Abstract Improved and modular tools are needed for the neuroanatomical dissection of CNS axonal tracts, and to study the cell-intrinsic and cell-extrinsic cues that govern their assembly and plasticity. Here we describe a general purpose transgenic tracer that can be used to visualize axonal tracts and synaptic terminals in any region of the embryonic neural tube or postnatal CNS, on any wild type or mutant genetic background. The construct permits CRE-inducible expression of a dicistronic axonal marker encoding two surface reporter proteins: a farnesylated GFP and the human Placental Alkaline Phosphatase (PLAP). Both proteins localize alongside the neuronal surface, permitting the concomitant detection of cell body, neurites, and presynaptic and postsynaptic sites in the same neuron. This provides a CRE-inducible dual system for imaging neural circuits in vivo, and to study their assembly and remodeling in cultured neurons, neural stem cells, and tissue explants derived from the reporter line. Unlike existing lines, this reporter does not encode a ubiquitously expressed, floxable LacZ gene, permitting the simultaneous analysis of beta galactosidase activity in mutant lines. genesis 45:405,412, 2007. © 2007 Wiley-Liss, Inc. [source] Combined Pressure,temperature Inactivation of Alkaline Phosphatase in Bovine Milk: A Kinetic StudyJOURNAL OF FOOD SCIENCE, Issue 1 2000L. Ludikhuyze ABSTRACT: A detailed kinetic study on pressure-temperature inactivation of alkaline phosphatase has been performed in the pressure range 0.1 to 725 MPa at temperatures between 25 and 63 °C. Inactivation could be accurately described by a first order kinetic model, allowing D-values to be calculated. According to the thermal death time terminology, zr - and zp -values were calculated, expressing temperature and pressure dependence respectively. However, at high temperature, pressure dependence could not be calculated unambiguously. D-values firstly increased with increasing pressure up to 300 MPa and then decreased with further pressure increase, showing thermal inactivation to be counteracted by low pressure. Finally, a global model describing the D-value as a function of pressure and temperature has been formulated. [source] Effects of Nigella orientalis and N. segetalis fixed oils on blood biochemistry in ratsPHYTOTHERAPY RESEARCH, Issue 1 2006G. Kökdil Abstract Nigella orientalis and N. segetalis fixed oils were administered orally (1 mL/kg/day) to Wistar Kyoto rats for 4 weeks. The effects of the oils on biochemical parameters were compared with a control group that received distilled water under identical conditions. LDL-cholesterol level was decreased significantly in both oil groups while serum total cholesterol and VLDL-cholesterol were decreased significantly following administration of only N. orientalis fixed oil when compared with the control group. The HDL-cholesterol levels were increased significantly in both oil groups. N. orientalis fixed oil significantly reduced Aspartateaminotransferase (AST), Alkaline Phosphatase (ALP), bilirubin and urea levels in rats. There was an increase in the albumin, uric acid and mean corpuscular volume (MCV) concentrations, while the mean corpuscular hemoglobin concentration (MCHC) and RDW (red cell distribution width) levels decreased significantly. In N. segetalis fixed oil treated rats, the levels of ALP, Blood Urea Nitrogen (BUN), MCHC, RDW were decreased significantly, whereas a significant increase was found in albumin, fibrinogen, Hematocrit (HCT) and MCV levels. The effects of 4 weeks oral intake of N. orientalis and N. segetalis fixed oils on blood malondialdehyde (MDA) and total antioxidant status (TOS) were also investigated in rats. The study showed that the oils had no significant effect on MDA production. N. orientalis and N. segetalis fixed oils caused a significant increase in the total antioxidant status in rats. Copyright © 2006 John Wiley & Sons, Ltd. [source] Purification and Concentration of Alkaline Phosphatase by Selective Permeabilization of Escherichia coli Using Reverse Micellar SolutionsBIOTECHNOLOGY PROGRESS, Issue 6 2003Ritu Bansal-Mutalik Recovery of alkaline phosphatase (AP) from the periplasm of Escherichia coli using reverse micellar solutions (RMSs) of sodium dioctyl sulfosuccinate (AOT) in aliphatic hydrocarbons has been attempted. A variety of surface-active agents, solvents, and reverse micellar conditions were screened, and an excellent recovery of the enzyme in a concentrated form, with a high purification factor, was obtained in a single-step process. The permeabilization process strongly depended on the water content of the RMS as well as on the amount of water coating the microbial cell surface. The product was almost free from nucleic acids. In addition, because of the low affinity of AOT and the organic solvent for the aqueous phase, contamination by the permeabilizing agents would also be negligible. [source] A New 4.0-Generation Dendrimer Phosphorescence Labeling Reagent and Its Application to Determination of Trace Alkaline Phosphatase by Affinity Adsorption Solid Substrate-room Temperature PhosphorimetryCHINESE JOURNAL OF CHEMISTRY, Issue 10 2007Zhi-Ming LI Abstract A Triton X-100-4.0G-D (4.0G-D refers to a 4.0-generation dendrimer) was brought forward as a new phosphorescence labeling reagent. Two types of specific affinity adsorption (AA) reactions (direct method and sandwich method) were carried out between the labeling product of Triton X-100-4.0G-D-Wheat germ agglutinin (WGA) and alkaline phosphatase (ALP), the product of AA reaction preserved the good characteristics of room temperature phosphorescence (RTP) of 4.0G-D and ,IP of the product was proportional to the content of ALP. According to the fact stated above, a new method for the determination of trace ALP by affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) was established on the basis of WGA labeled with the Triton X-100-4.0G-D. The detection limits were 0.20 ag·spot,1 (corresponding concentration: 5.0×10,16 g·mL,1, namely 5.0×10,18 mol·L,1) for a direct method and 0.14 ag·spot,1 (corresponding concentration: 3.5×10,16 g·mL,1, namely 3.5×10,18 mol·L,1) for a sandwich method, respectively. For their high sensitivity, good repeatability and high accuracy, the direct method and sandwich method have been successfully applied to determine the content of ALP in human serum, and the results were coincided with the clinical detection results of the enzyme-linked immunosorbent assay method by the Zhangzhou Hospital of Traditional Chinese Medicine. Meanwhile, the mechanism for the determination of trace ALP by AA-SS-RTP was discussed. [source] The regulation of osteogenesis by ECM rigidity in MC3T3-E1 cells requires MAPK activationJOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2007Chirag B. Khatiwala Once thought to provide only structural support to tissues by acting as a scaffold to which cells bind, it is now widely recognized that the extracellular matrix (ECM) provides instructive signals that dictate cell behavior. Recently we demonstrated that mechanical cues intrinsic to the ECM directly regulate the behavior of pre-osteoblastic MC3T3-E1 cells. We hypothesized that one possible mechanism by which ECM compliance exerts its influence on osteogenesis is by modulating the mitogen-activated protein kinase (MAPK) pathway. To address this hypothesis, the differentiation of MC3T3-E1 cells cultured on poly(ethylene glycol) (PEG)-based model substrates with tunable mechanical properties was assessed. Alkaline phosphatase (ALP) levels at days 7 and 14 were found to be significantly higher in cells grown on stiffer substrates (423.9 kPa hydrogels and rigid tissue culture polystyrene (TCPS) control) than on a soft hydrogel (13.7 kPa). Osteocalcin (OCN) and bone sialoprotein (BSP) gene expression levels followed a similar trend. In parallel, MAPK activity was significantly higher in cells cultured on stiffer substrates at both time points. Inhibiting this activation pharmacologically, using PD98059, resulted in significantly lower ALP levels, OCN, and BSP gene expression levels on the hydrogels. Interestingly, the effectiveness of PD98059 was itself dependent on substrate stiffness, with marked inhibition of MAPK phosphorylation in cells grown on compliant hydrogels but insignificant reduction in cells grown on TCPS. Together, these data confirm a role for MAPK in the regulation of osteogenic differentiation by ECM compliance. J. Cell. Physiol. 211: 661,672, 2007. © 2007 Wiley-Liss, Inc. [source] Alterations of liver function test in patients treated with antipsychoticsJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 6 2003M. Teresa Garcia-Unzueta Abstract The prevalence of alterations of liver function tests in patients treated with a wide range of antypsychotics is unknown. The aim of this study was to analyze the effects of antipsychotics on liver function tests in a population of schizophrenic outpatients. Concentrations of AST, ALT, GGT, alkaline phosphatase, albumin, and bilirubin were determined in 54 patients fitting DSM-IV criteria of schizophrenia, and the same number of sex- and age-matched healthy subjects. Assessments included the Clinical Global Impression (CGI) and the Positive and Negative Syndrome Scale (PANSS) in addition to treatment related variables. Transaminases concentrations were slightly elevated in study patients compared to healthy controls, but without statistical significance. Alkaline phosphatase showed higher values in schizophrenic patients. Albumin and bilirubin were lower in study patients. Liver function tests abnormalities were found in about 10% of schizophrenic patients treated with antipsychotics. Treatment with depot phenotiazines induces alteration in these tests more frequently than treatment with other antipsychotics. PANSS negative subscale scores directly correlated with alkaline phosphatase and inversely correlated with albumin. A substantial number of patients in treatment with antipsychotic drugs present alterations of liver function tests. Both pharmacological and clinical factors could be related with these alterations. J. Clin. Lab. Anal. 17:216,218, 2003. © 2003 Wiley-Liss, Inc. [source] Pressureless Sintering and Mechanical and Biological Properties of Fluor-hydroxyapatite Composites with ZirconiaJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 12 2003Hae-Won Kim Fluor-hydroxyapatite (FHA) fabricated by a reaction between fluorapatite (FA) and hydroxyapatite (HA) was mixed with ZrO2 to produce FHA,ZrO2 composites. When the relative amount of FA to HA increased, the decomposition of the composite was decreased gradually because of the formation of thermally stable FHA solid solutions. With such suppression of decomposition, the FHA,ZrO2 composites retained fully densified bodies. As a result, significant enhancements in mechanical properties, such as hardness, flexural strength, and fracture toughness, were achieved as the relative amount of FA to HA increased. The highest values in strength and toughness were 220 MPa and 2.5 MPa·m1/2, respectively, with FHA,40 vol% ZrO2 composites. In vitro proliferation of osteoblast-like cells (MG63) on the composites showed behavior similar to that observed on pure HA and FHA. Alkaline phosphatase (ALP) activity of the growing cells (HOS) on the composites was slightly down-regulated compared with that on pure HA and FHA at prolonged periods. [source] Kinetic behaviour and stability of Escherichia coli ATCC27257 alkaline phosphatase immobilised in soil humatesJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2003María C Pilar Abstract Alkaline phosphatase (EC 3.1.3.1) extracted from Escherichia coli ATCC27257 was immobilised by co-flocculation with soil humates in the presence of Ca2+. The effects of time, temperature, pH and concentration of enzyme and support on immobilisation were studied. Between 58 and 92% of the added phosphatase was strongly bound to the humates, depending on the conditions of immobilisation used. Some characteristics of the humate,phosphatase complexes and of the free enzyme were compared. The enzymatic complexes showed values of Km (2.22,mM) and activation energy (33.4,kJ,mol,1) similar to those of the free enzyme (2.00,mM and 27.6,kJ,mol,1). The pH/activity profiles revealed no change in terms of shape or optimum pH (10.5) upon immobilisation of alkaline phosphatase. However, the immobilised enzyme showed maximal activity in the range of 80,100,°C, while the free enzyme had its highest activity at 60,°C. The thermal stability of alkaline phosphatase was enhanced by complexation to the soil humates. © 2003 Society of Chemical Industry [source] Generation and Characterization of Embryonic Stem-Like Cell Lines Derived from In Vitro Fertilization Buffalo (Bubalus bubalis) EmbryosREPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2010B Huang Contents In the present study, buffalo embryonic stem-like (ES-like) cell lines were successfully isolated, cultured and characterized. From a total of 92 normal buffalo embryos obtained by in vitro fertilization, 18 were morulae, 33 were blastocyst and 41 were hatched blastocyst, the inside of morulae or inner cell masses of blastocysts were isolated mechanically and cultured onto mitomocin-C-inactivated buffalo embryonic fibroblasts as feeder layers. Alkaline phosphatase (AP) of ES-like cells, as well as the specific stage embryonic antigen SSEA-1, SSEA-3, SSEA-4 and transcription factor OCT-4, was used to evaluate the characterization of the cells. The spontaneous differentiation of ES-like cells was induced by culturing on leukaemia inhibitory factor-free medium for more than 2 weeks without passage. To evaluate mark gene expression, total RNA was extracted from cells, and specific primers were used for reverse transcriptase-polymerase chain reaction (RT-PCR). After 8,10 days of culture, primary ES-like cell colonies were formed in 0% (0/18) of morulae, 24.24% (8/33) of blastocysts and 60.98% (25/41) of hatched blastocysts, respectively. The forming rate of primary ES-like cells colonies in hatched blastocyst group was significantly (p < 0.05) higher than the obtained for other groups. Two ES-like cell lines could survive to eight passages at least by using the method of mechanical dissociation, but just three passages by using the method of enzymatic dissociation. The cells formed large, multicellular colonies with distinct boundaries, exhibited many important features of ES/ES-like cells, including positive AP, SSEA-1, SSEA-3 and SSEA-4 activity. Undifferentiated buffalo ES-like cells expressed Oct-4, Nanog, Sox2 gene mRNA. In vitro differentiation experiments had demonstrated that those cells were pluripotent. [source] Study of the Structure of Canine Mesenchymal Stem Cell Osteogenic CultureANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 5 2010M. B. Eslaminejad With 6 figures and 1 table Summary This study was designed to investigate the morphological features of osteogenic cultures that were established from canine marrow derived-mesenchymal stem cells (MSCs). Tripotent canine MSCs were plated in osteogenic conditions for 3 weeks, at the end of which the cultures were observed by light and transmission electron microscopy. Alkaline phosphatase (ALP) activity of the culture was determined during the differentiation period. To assess whether endochondral or intramembranous ossification was involved in MSC bone differentiation, the cultures were explored for cartilage-related gene expression. Multiple nodule-like cell aggregates appeared to form in the osteogenic cultures. These nodules were covered by a periosteum-like layer and osteocyte-like cells of varying morphology were located in lacuna-like cavities within the nodule mass. Furthermore, the bone nodules possessed an abundant matrix in which clearly striated collagen I fibres were arranged in perpendicular bundles. Matrix vesicles involving in matrix mineralization were evident in the nodules. This was in accordance with increased ALP activity in the culture. No expression of cartilage-related genes was observed, which suggested that osteogenesis might occur by intramembranous ossification. In conclusion, canine MSCs could be an appropriate model for studying in vitro bone development. [source] Bioavailability of amino acids chelated and glass embedded zinc to rainbow trout, Oncorhynchus mykiss, fingerlingsAQUACULTURE NUTRITION, Issue 4 2001M.J. Apines A 2 × 4 factorial experiment was conducted to determine the bioavailability of zinc (Zn) from amino acids chelated (Zn,Am) and glass embedded Zn (Zn,Gl) as sources for rainbow trout, Oncorhynchus mykiss, fed practical type diets. Two levels of Zn (20 and 40 mg kg,1) were supplemented to the diets using either zinc sulphate (Zn,Sf), zinc methionine (Zn,Mt), Zn,Am or Zn,Gl. Rainbow trout with an average weight of 2 g were fed the experimental diets for 15 weeks. Growth and feed gain ratio (FGR) were not significantly influenced by the dietary Zn content and forms. Alkaline phosphatase (ALP) activity for both levels of Zn,Am was significantly higher than that of Zn,Sf and Zn,Gl at 20 mg supplementation. In another experiment, fish of about 95 g were fed the same experimental diets to determine the absorption of Zn and it was found to be significantly higher from Zn,Am compared with the rest. Retention from Zn,Am at 20 mg was significantly higher than the rest, excluding Zn,Sf. The results suggest that the availability of Zn from Zn,Am might be superior among the sources compared. [source] Growth, digestive capacity and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of dietary inositolAQUACULTURE RESEARCH, Issue 8 2009Wei-Dan Jiang Abstract A 60-day feeding trial was carried out with juvenile Jian carp (Cyprinus carpio var. Jian) to study the effects of myo -inositol (MI) on the growth, digestive enzyme and intestinal microbial population. Diets with seven levels of inositol (163.5, 232.7, 384.2, 535.8, 687.3, 838.8 and 990.3 mg MI kg,1 diet) were fed to Jian carp (initial weight 22.28±0.07 g). Per cent weight gain (PWG) was improved with increasing inositol levels up to 535.8 mg MI kg,1 diet (P<0.05), and plateaued (P>0.05). The protein production value, lipid production value and ash production value were increased with increasing dietary inositol levels up to 384.2, 838.8 and 838.8 mg MI kg,1 diet respectively (P<0.05). Although intestinal protein content and trypsin activity were not affected by inositol levels (P>0.05), chymotrypsin, lipase and amylase activities in intestine were the lowest for fish fed the MI-unsupplemented diet (P<0.05). Alkaline phosphatase, Na+, K+ -ATPase, ,-glutamyl transpeptidase and creatinkinase activities in the intestine were increased with an increase in the inositol levels up to 384.2,687.3 mg MI kg,1 diet (P<0.05). Intestinal Aeromonas hydrophila and Escherichia coli decreased with an increase in the levels of dietary inositol up to 232.7 and 687.3 mg MI kg,1 diet respectively (P<0.05), while Lactobacillus in the intestine increased with an increase in inositol levels up to 990.3 mg MI kg,1 diet (P<0.05). In conclusion, inositol improved growth, digestive capacity and intestinal microbial population of juvenile Jian carp, and the dietary inositol requirement for PWG of juvenile Jian carp is 518.0 mg MI kg,1 diet. [source] Altered mineralization of human osteoarthritic osteoblasts is attributable to abnormal type I collagen productionARTHRITIS & RHEUMATISM, Issue 5 2009Denis Couchourel Objective Bone tissue in osteoarthritis (OA) is composed of abundant undermineralized osteoid matrix. The aim of this study was to investigate the mechanisms responsible for this abnormal matrix, using in vitro OA subchondral osteoblasts. Methods Primary normal and OA osteoblasts were prepared from tibial plateaus. Phenotype was determined by alkaline phosphatase activity, and osteocalcin, osteopontin, prostaglandin E2 (PGE2), and transforming growth factor ,1 (TGF,1) were assessed by enzyme-linked immunosorbent assay. Expression of COL1A1 and COL1A2 was determined by real-time polymerase chain reaction. The production of type I collagen was determined by the release of its C-terminal propeptide and Western blot analysis. In vitro mineralization was evaluated by alizarin red staining. Inhibition of TGF,1 expression was performed using a small interfering RNA technique. Results Mineralization of OA osteoblasts was reduced compared with mineralization of normal osteoblasts, even in the presence of bone morphogenetic protein 2 (BMP-2). Alkaline phosphatase and osteocalcin levels were elevated in OA osteoblasts compared with normal osteoblasts, whereas osteopontin levels were similar. The COL1A1 -to- COL1A2 messenger RNA ratio was 3-fold higher in OA osteoblasts compared with normal osteoblasts, and the production of collagen by OA osteoblasts was increased. Because TGF,1 inhibits BMP-2,dependent mineralization, and because TGF,1 levels are ,4-fold higher in OA osteoblasts than in normal osteoblasts, inhibiting TGF,1 levels in OA osteoblasts corrected the abnormal COL1A1 -to- COL1A2 ratio and increased alizarin red staining. Conclusion Elevated TGF,1 levels in OA osteoblasts are responsible, in part, for the abnormal ratio of COL1A1 to COL1A2 and for the abnormal production of mature type I collagen. This abnormal COL1A1 -to- COL1A2 ratio generates a matrix that blunts mineralization in OA osteoblasts. [source] Crystallization and preliminary X-ray crystallographic analysis of PhoK, an extracellular alkaline phosphatase from Sphingomonas sp.ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009BSAR- Alkaline phosphatases (APs) are widely distributed from microbes to humans and are involved in several important biological processes such as phosphate nutrition, signal transduction and pathogenesis. Alkaline phosphatases are also useful in various industrial applications and in recombinant DNA technology. A new AP enzyme from Sphingomonas sp. strain BSAR-1, termed PhoK, has been shown to be useful in uranium bioprecipitation. PhoK was expressed, purified and crystallized. The crystals belonged to space group P43212 or P41212, with unit-cell parameters a = b = 87.37, c = 168.16,Å, and contained one enzyme molecule in the asymmetric unit. Native diffraction data have been collected to 1.95,Å resolution at the ESRF. [source] Effect of single and binary combinations of plant-derived molluscicides on different enzyme activities in the nervous tissue of Achatina fulicaJOURNAL OF APPLIED TOXICOLOGY, Issue 1 2003I. G. Rao Abstract Effect of single and binary treatments of plant-derived molluscicides on different enzymes,acetylcholinesterase (AChE), lactic dehydrogenase (LDH) and acid/alkaline phosphatase (ACP/ALP),in the nervous tissue of the harmful terrestrial snail Achatina fulica were studied. Sublethal in vivo 24-h exposure to 40% and 80% LC50 of Azadirachta indica oil, Cedrus deodara oil, Allium sativum bulb powder, Nerium indicum bark powder and binary combinations of A. sativum (AS) + C. deodara (CD) and CD + A. indica (AI) oils significantly altered the activity of these enzymes in the nervous tissue of Achatina fulica. The binary treatment of AS + CD was more effective against AChE, LDH, and ALP than the single ones. However, binary treatment of AI + CD was more effective against ALP. Copyright © 2003 John Wiley & Sons, Ltd. [source] Pamidronate treatment of bone fibrous dysplasia in nine children with McCune-Albright syndromeACTA PAEDIATRICA, Issue 2 2000R Lala McCune-Albright syndrome is a rare genetic disorder consisting of skin and bone dysplasia and peripheral endocrinopathies. Little data have been collected regarding bisphosphonate treatment of bone fibrous dysplasia in paediatric patients with this syndrome. The aim of our study was to investigate the therapeutic efficacy of pamidronate in these patients. Nine patients with moderate to severe forms of bone fibrous dysplasia were treated with pamidronate intravenously (0.5-1 mg/ kg/daily for 2-3 d) at 0.5-1-y intervals. Patients were treated over a time period of 0.5-3.5 y. During treatment no spontaneous fracture occurred. Bone pain and gait abnormality due to pain disappeared after 2-3 therapeutic cycles. Cranial asymmetry and limb length discrepancy remained unchanged. Elevated serum alkaline phosphatase and urine hydroxyproline values were reduced by the treatment, demonstrating drug activity at the lesional level. The effectiveness of pamidronate was also seen at the non-lesional level through an increase in bone density. Radiographic and scintigraphic evidence of lesion healing was not attained. Pamidronate treatment can ameliorate the course of bone fibrous dysplasia in children and adolescents with McCune-Albright syndrome. [source] Optimization of a flow cytometry-based protocol for detection and phenotypic characterization of multipotent mesenchymal stromal cells from human bone marrowCYTOMETRY, Issue 6 2006Elena A. Jones Abstract Background: To study the biology of rare bone marrow (BM) multipotent mesenchymal stromal cells (MSCs), recognized protocols are needed. Colony-forming unit-fibroblast (CFU-F) assays have historically been used for the enumeration of MSCs. However, the need to isolate and further analyze MSCs requires new strategies based on cell surface markers. The purpose of this work was to verify the phenotype of BM MSCs in vivo and to develop flow cytometry-based methods for their evaluation. Methods: Pre-enrichment with D7-FIB-conjugated microbeads, cell sorting for CD45lowD7-FIB+LNGFR+ cells, and CFU-F assay were used to confirm the phenotype of BM MSCs in vivo. Further phenotypic characterization of MSCs was performed using three-color flow cytometry following pre-enrichment or by direct four-color flow cytometry. The sensitivity of direct flow cytometry/rare event analysis for the accurate enumeration of MSCs was validated using 85 samples from patients with neoplastic BM diseases. Results: In normal BM, a significant correlation was found between the frequencies of CFU-Fs and CD45lowD7-FIB+LNGFR+ cells (n = 19, R = 0.719, P = 0.001). Following cell sorting, ,15% of these cells were clonogenic. The same cells were enriched using LNGFR-based positive selection, CD45/Glycophorin A-based depletion, or plastic adherence. CD45lowD7-FIB+LNGFR+ cells expressed classic makers of cultured MSCs CD73/SH3 and CD105/SH2 and markers of stromal reticular cells CD106/VCAM and alkaline phosphatase. Novel markers were identified including leukemia inhibitory factor receptor and gp130. CD45lowD7-FIB+LNGFR+ cells were increased fourfold in the floating fat fraction of normal BM aspirates. Their frequency was decreased in chronic lymphocytic leukemia (threefold, n = 13, P = 0.049) and chronic myelogenous leukemia (ninefold, n = 11, P = 0.001) compared with that in age-matched controls (n = 26 and n = 31, respectively). Conclusions: This study demonstrates the usefulness of flow cytometry-based methods for the detection, enumeration and further phenotypic analysis of BM MSCs. These findings have broad applications for the future evaluation of BM MSCs in health and disease. © 2006 International Society for Analytical Cytology [source] HMG-CoA reductase inhibitors prevent bone loss in patients with Type 2 diabetes mellitusDIABETIC MEDICINE, Issue 9 2004A. Nakashima Abstract Aims It has been reported that 3-hydroxy-3-methylglutaryl co-enzyme A (HMG-CoA) reductase inhibitors increase bone mineral density (BMD) in vivo. We investigated the effect of HMG-CoA reductase inhibitors on BMD in patients with Type 2 diabetes mellitus. Patients and methods We selected 122 patients with Type 2 diabetes, who were not taking active vitamin D preparations. Their mean age was 67.3 ± 9.2 years. They were divided into a control group (n = 63) without HMG-CoA reductase inhibitor therapy and an HMG-CoA group (n = 59) who were treated with these drugs. The BMD of the distal one-third of the radius was measured by dual-energy X-ray adsorptiometry at baseline and after 2 years. Results There were no significant differences between the control and HMG-CoA groups at baseline with respect to age, gender, body mass index, duration of diabetes, haemoglobin A1c, fasting plasma glucose, adjusted calcium, serum phosphorus, alkaline phosphatase, albumin excretion rate and radial BMD. However, there was a significantly smaller annual decrease of the radial BMD in the HMG-CoA group. Multiple regression analysis with a forward elimination procedure revealed a positive correlation of the radial BMD Z-score with body mass index, while there was a negative correlation with alkaline phosphatase and albumin excretion rate. In addition, the annual rate of change of the radial BMD showed a positive correlation with HMG-CoA reductase inhibitor therapy. Conclusions These findings suggest that HMG-CoA reductase inhibitors may prevent bone loss in patients with Type 2 diabetes. [source] Epithelioid angiosarcoma: A neoplasm with potential diagnostic challengesDIAGNOSTIC CYTOPATHOLOGY, Issue 2 2010Christine F. Lin B.S. (Student) Abstract Epithelioid angiosarcomas are extremely rare tumors associated with poor prognosis and early metastases. Its epithelioid cytomorphology and limited vasoformation make it difficult to distinguish from more common malignancies, such as, carcinoma. This can be a potential diagnostic pitfall for the cytopathologist. In this report, the patient is a 24-year-old man presenting with testicular pain, a pelvic mass, and innumerable liver nodules. Immediate interpretation of the needle core biopsies of the pelvic mass and liver lesions initially favored a poorly differentiated adenocarcinoma. Unusual positive immunohistochemical stains for CD30 and CK7 ultimately led the investigation toward a tumor of mesenchymal origin. Further, immunohistochemical evaluation demonstrated positive CD31 and Factor VIII staining and established the final diagnosis of epithelioid angiosarcoma. The tumor cells were negative for CD34, CK20, alpha-fetoprotein, placental-like alkaline phosphatase, hepatocyte paraffin 1, polyclonal carcinoembryonic antigen, CD10, CA-125, prostate-specific antigen, and prostatic acid phosphatase. This case is reported to illustrate the importance of considering the diagnosis of epithelioid angiosarcoma when encountering an "epithelioid" neoplasm particularly with unusual immunoreactivity for CK7 and CD30. Diagn. Cytopathol. 2010. © 2009 Wiley-Liss, Inc. [source] Platform for Highly Sensitive Alkaline Phosphatase-Based Immunosensors Using 1-Naphthyl Phosphate and an Avidin-Modified Indium Tin Oxide ElectrodeELECTROANALYSIS, Issue 19 2009Abdul Aziz Abstract We report a versatile platform for highly sensitive alkaline phosphatase (ALP)-based electrochemical biosensors that uses an avidin-modified indium tin oxide (ITO) electrode as a sensing electrode and 1-naphthyl phosphate (NPP) as an ALP substrate. Almost no electrocatalytic activity of NPP and good electrocatalytic activity of 1-naphthol (ALP product) on the ITO electrodes allow a high signal-to-background ratio. The effective surface covering of avidin on the ITO electrodes allows very low levels of nonspecific binding of proteins to the sensing electrodes. The platform technology is used to detect mouse IgG with a detection limit of 1.0,pg/mL. [source] Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich AssayELECTROANALYSIS, Issue 11 2009Sonia Centi Abstract A disposable electrochemical assay involving magnetic particles and carbon-based screen-printed electrodes (SPCEs) was developed for the detection of C Reactive Protein (CRP). CRP is a plasma protein and is among the most expressed proteins in acute phase inflammation cases, being a known biomarker for inflammatory states. The assay was based on a sandwich format in which a RNA aptamer was coupled to a monoclonal antibody and alkaline phosphatase (AP) was used as enzymatic label. After the sandwich assay, the modified magnetic beads were captured by a magnet on the surface of a graphite working electrode and the electrochemical detection was thus achieved through the addition of the AP substrate (,-naphthyl-phosphate) and ,-naphthol produced during the enzymatic reaction was detected using differential pulse voltammetry (DPV). The parameters influencing the different steps of the assay were optimized in order to reach the best sensitivity and specificity. With the optimized conditions, the assay was applied to the analysis of CRP free serum and serum samples. [source] Comparison of Electrochemical and Surface Plasmon Resonance Immunosensor Responses on Single Thin FilmELECTROANALYSIS, Issue 20 2008Ryoji Kurita Abstract This paper reports results obtained when comparing an electrochemical enzyme immunosensor and a surface plasmon resonance (SPR) based immunosensor on the same gold surface installed in an electrochemical SPR flow cell. Simultaneous electrochemical and SPR measurements were performed on a gold surface modified with multilayers of poly- L -lysine and poly-styrenesulfonate assembled with the layer-by-layer method. First, we obtained the SPR response induced by the formation of an immunocomplex from the shift in the SPR angle by injecting an anti tumor necrosis factor-, antibody solution labeled with alkaline phosphatase into the flow cell containing the multilayer modified with tumor necrosis factor-,. Then we compared this SPR result with that obtained for the electrochemical oxidation current of p -aminophenol catalyzed by alkaline phosphatase from p -aminophenolphosphate on the same gold film. We compared the two immunosensor responses obtained using the different measurement principles and found that there was a high correlation efficient of 0.973 between them. This was because we were able to immobilize the immunoreagents with good stability and without losing the transport of the enzyme product in the multilayer whose thickness we easily controlled with nanometer scale accuracy. We also report that the detection limit of our electrochemical immunosensor after optimization was around 100,pg/mL (0.4,pM), which is one of the lowest values yet reported for an electrochemical immunosensor. [source] Development of a Rapid Single-Drop Analysis Biosensor for Screening of Phenanthrene in Water SamplesELECTROANALYSIS, Issue 20 2004Abstract Detection techniques for biosensors often require bulky instruments or cells that are not feasible for in-field analysis. Our single-drop cell design, optimized in this work, comprised a screen-printed three-electrode (SPE), strip in horizontal position onto which a volume of 100,,L of sample or substrate solution was placed to ensure electrical contact (complete circuit). Together with optimized linear sweep voltammetry (LSV), parameters for the detection of the enzyme alkaline phosphatase (AP), the system was applied to a biosensor for the analysis of polycyclic aromatic hydrocarbons (PAHs), in environmental samples. A limit of detection (LOD), of 0.15,ppb was achieved for a model system with an IC50 value of 0.885 ppb and a linear range (LR), of 0.2,10,ppb. Application of the single drop analysis (SDA), format to a PAH biosensor gave a LOD of 1.4,ppb for detection of phenanthrene with an IC50 value of 29.3,ppb and linear range of 2,100,ppb. Proof of concept is shown with spiked sample analysis of phenanthrene in matrices such as sea, river and tap water. [source] Microfluidic tectonics platform: A colorimetric, disposable botulinum toxin enzyme-linked immunosorbent assay systemELECTROPHORESIS, Issue 10-11 2004Jaisree Moorthy Abstract A fabrication platform for realizing integrated microfluidic devices is discussed. The platform allows for creating specific microsystems for multistep assays in an ad hoc manner as the components that perform the assay steps can be created at any location inside the device via in situ fabrication. The platform was utilized to create a prototype microsystem for detecting botulinum neurotoxin directly from whole blood. Process steps such as sample preparation by filtration, mixing and incubation with reagents was carried out on the device. Various microfluidic components such as channel network, valves and porous filter were fabricated from prepolymer mixture consisting of monomer, cross-linker and a photoinitiator. For detection of the toxoid, biotinylated antibodies were immobilized on streptavidin-functionalized agarose gel beads. The gel beads were introduced into the device and were used as readouts. Enzymatic reaction between alkaline phosphatase (on secondary antibody) and substrate produced an insoluble, colored precipitate that coated the beads thus making the readout visible to the naked eye. Clinically relevant amounts of the toxin can be detected from whole blood using the portable enzyme-linked immunosorbent assay (ELISA) system. Multiple layers can be realized for effective space utilization and creating a three-dimensional (3-D) chaotic mixer. In addition, external materials such as membranes can be incorporated into the device as components. Individual components that were necessary to perform these steps were characterized, and their mutual compatibility is also discussed. [source] Hematotoxic and hepatotoxic effects of dichlorvos at sublethal dosages in ratsENVIRONMENTAL TOXICOLOGY, Issue 2 2009Ismail Celik Abstract The present study was designed to understand the effects of sublethal concentrations of dichlorvos (DIC) on hematological constituent [red blood corpuscles, white blood corpuscles (WBC), mean cell volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelet counts, hemoglobin and hematocrite levels] and serum damage marker enzymes (aspartate aminotransferase, alanin aminotransferase, alkaline phosphatase, and lactate dehydrogenase) in rats at subacute period under laboratory conditions. DIC at dosages of 5 and 10 ppm was administered orally to six male rats ad libitum during the tests for 4 weeks consecutively. According to the results, DIC treatments increased significantly the levels of serum marker enzyme activities, whereas they did not change hematologic constituent except for WBC number treated with both dosages of DIC. The observations presented led us to conclude that the administrations of subacute DIC induced the levels of damage marker enzymes and leukocytosis. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009. [source] Mouse toxicity of Anabaena flos-aquae from Lake Dianchi, ChinaENVIRONMENTAL TOXICOLOGY, Issue 1 2009Xiaojie Pan Abstract Some species of the genera Anabaena can produce various kinds of cyanotoxins, which may pose risks to environment and human health. Anabaena has frequently been observed in eutrophic freshwater of China in recent years, but its toxicity has been reported only in a few studies. In the present study, the toxicity of an Anabaena flos-aquae strain isolated from Lake Dianchi was investigated. Acute toxicity testing was performed by mouse bioassay using crude extracts from the lyophilized cultures. The mice exposed to crude extracts showed visible symptoms of toxicity and died within 10,24 h of the injection. Serum biochemical parameters were evaluated by the use of commercial diagnostic kits. Significant alterations were found in the serum biochemical parameters: alkaline phosphatase (AKP), ,-glutamyl transpeptidase (,-GT), aspartate amino transferase (AST), alanine amino transferase (ALT), AST/ALT ratio, total protein content, albumin content, albumin/globulin (A/G) ratio, blood urea nitrogen (BUN), serum creatinine (Ssr), and total antioxidative capacity (T-AOC). Histopathological observations were carried out with hematoxylin and eosin (HE) stain under light microscope. Severe lesions were seen in the livers, kidneys, and lungs of the mice injected with crude extracts. The alterations of biochemical parameters were in a dose-dependent manner, and the severities of histological lesions were in the same manner. Based on biochemical and histological studies, this research firstly shows the presence of toxin-producing Anabaena species in Lake Dianchi and the toxic effects of its crude extracts on mammals. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009. [source] Levels of transaminases, alkaline phosphatase, and protein in tissues of Clarias gariepienus fingerlings exposed to sublethal concentrations of cadmium chlorideENVIRONMENTAL TOXICOLOGY, Issue 6 2008Babu Velmurugan Abstract The freshwater fish, Clarias gariepienus fingerlings, were exposed to sublethal concentrations (1.7 and 3.4 mg/L) of cadmium chloride for 12 days. Aspartate aminotransferase (AAT), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total protein levels were assayed in the gill, brain, and muscle of the fish at regular intervals of 6 and 12 days. The activities of AAT, ALT, and ALP of the treated fishes increased significantly in all the tissues compared with the control fish. Protein level in all the tissues showed a significant decrease in comparison to unexposed controls throughout the experimental periods. These results revealed that cadmium chloride effects the intermediary metabolism of C. gariepienus fingerlings and that the assayed enzymes can work as good biomarkers of contamination. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source] | ||||||||||||||||||||||||||||||||||||||||