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Sample records for calcium pyrophosphate dihydrate

  1. Pseudotumor of temporomandibular joint: destructive calcium pyrophosphate dihydrate arthropathy.

    PubMed

    Pritzker, K P; Phillips, H; Luk, S C; Koven, I H; Kiss, A; Houpt, J B

    1976-03-01

    The clinical and pathological features of a tumor of the temporamandibular joint occurring in a 55 year old man, and subsequently identified as a calcium pyrophosphate dihydrate (CPPD) arthropathy, are reported. Crystalline deposits were identified by compensated light microscopy and confirmed with X-ray diffraction, transmission, and scanning electron microscopy. The relationship of this unique case to other clinical presentations of CPPD deposition disease and the implications of the histological features to the pathogenesis of pseudogout are discussed. This case demonstrates that CPPD arthropathy should be included in the differential diagnosis of an arthrosis or of a tumor involving the temporomandibular joint.

  2. The ANKH gene and familial calcium pyrophosphate dihydrate deposition disease.

    PubMed

    Netter, Patrick; Bardin, Thomas; Bianchi, Arnaud; Richette, Pascal; Loeuille, Damien

    2004-09-01

    Familial calcium pyrophosphate dihydrate deposition (CPPD) disease is a chronic condition in which CPPD microcrystals deposit in the joint fluid, cartilage, and periarticular tissues. Two forms of familial CPPD disease have been identified: CCAL1 and CCAL2. The CCAL1 locus is located on the long arm of chromosome 8 and is associated with CPPD and severe osteoarthritis. The CCAL2 locus has been mapped to the short arm of chromosome 5 and identified in families from the Alsace region of France and the United Kingdom. The ANKH protein is involved in pyrophosphate metabolism and, more specifically, in pyrophosphate transport from the intracellular to the extracellular compartment. Numerous ANKH gene mutations cause familial CCAL2; they enhance ANKH protein activity, thereby elevating extracellular pyrophosphate levels and promoting the formation of pyrophosphate crystals, which produce the manifestations of the disease. Recent studies show that growth factors and cytokines can modify the expression of the normal ANKH protein. These results suggest a role for ANKH in sporadic CPPD disease and in CPPD associated with degenerative disease.

  3. Calcium pyrophosphate dihydrate crystal deposition of multiple lumbar facet joints: a case report.

    PubMed

    Namazie, Mohamed Ridzwan bin Mohamed; Fosbender, Murray R

    2012-08-01

    Pseudogout of the lumbar facet joints is rare. We report on a 69-year-old woman with 2-level symptomatic synovial cysts of the facet joints caused by calcium pyrophosphate dihydrate crystal deposition. She underwent surgical decompression for sciatica and low back pain. At one-year follow-up, she had recovered completely.

  4. Calcium pyrophosphate dihydrate deposition disease (CPPD)/Pseudogout of the temporomandibular joint - FNA findings and microanalysis.

    PubMed

    Naqvi, Asghar H; Abraham, Jerrold L; Kellman, Robert M; Khurana, Kamal K

    2008-04-21

    We report a case of a Calcium pyrophosphate dihydrate deposition disease (CPPD) presenting as a mass in the parotid and temporomandibular joint (TMJ) that simulated a parotid tumor. A 35 year-old man presented with pain in the left ear area. A CT Scan of the area showed a large, calcified mass surrounding the left condylar head, and extending into the infratemporal fossa. FNA of the mass showed birefringent crystals, most of which were rhomboid with occasional ones being needle shaped, embedded in an amorphous pink substance. Scanning electron microscopy (SEM) with energy dispersive x-ray spectroscopy (EDS) of these crystals showed peaks corresponding to calcium and phosphorus. SEM/EDS is a rapid method of diagnosing calcium pyrophosphate dihydrate deposition disease (CPPD) and an alternative to more commonly used method of special staining of cell block sections coupled with polarizing microscopy.

  5. Crystal structure of monoclinic calcium pyrophosphate dihydrate (m-CPPD) involved in inflammatory reactions and osteoarthritis.

    PubMed

    Gras, Pierre; Rey, Christian; André, Gilles; Charvillat, Cédric; Sarda, Stéphanie; Combes, Christèle

    2016-02-01

    Pure monoclinic calcium pyrophosphate dihydrate (m-CPPD) has been synthesized and characterized by synchrotron powder X-ray diffraction and neutron diffraction. Rietveld refinement of complementary diffraction data has, for the first time, allowed the crystal structure of m-CPPD to be solved. The monoclinic system P2(1)/n was confirmed and unit-cell parameters determined: a = 12.60842 (4), b = 9.24278 (4), c = 6.74885 (2) Å and β = 104.9916 (3)°. Neutron diffraction data especially have allowed the precise determination of the position of H atoms in the structure. The relationship between the m-CPPD crystal structure and that of the triclinic calcium pyrophosphate dihydrate (t-CPPD) phase as well as other pyrophosphate phases involving other divalent cations are discussed by considering the inflammatory potential of these phases and/or their involvement in different diseases. These original structural data represent a key step in the understanding of the mechanisms of crystal formation involved in different types of arthritis and to improve early detection of calcium pyrophosphate (CPP) phases in vivo.

  6. Synovial chondromatosis of the temporomandibular joint with calcium pyrophosphate dihydrate crystal deposition disease (pseudogout)

    PubMed Central

    Matsumura, Y; Nomura, J; Nakanishi, K; Yanase, S; Kato, H; Tagawa, T

    2012-01-01

    This report describes a very rare case of synovial chondromatosis with deposition of calcium pyrophosphate dihydrate (CPPD) crystals (pseudogout) in the temporomandibular joint (TMJ) of a 46-year-old male patient. Synovial chondromatosis is a non-neoplastic disease characterized by metaplasia of the connective tissue leading to chondrogenesis in the synovial membrane. Pseudogout is an inflammatory disease of the joints caused by the deposition of CPPD, producing similar symptoms to those observed in gout but not hyperuricaemia. Both diseases commonly affect the knee, hip and elbow joints, but rarely affect the TMJ. PMID:23166363

  7. [Calcium pyrophosphate dihydrate crystal deposition disease: a clinicopathologic analysis of 20 cases].

    PubMed

    Fang, Hui-qiong; Li, Qi-ming; Huang, Yao-qu; Xing, Ji-si; Mao, Rong-jun; Xie, Le

    2012-12-01

    To investigate the clinicopathologic features of calcium pyrophosphate dihydrate crystal deposition disease (CPPD-CDD). The clinical and pathologic profiles were retrospectively analysed in 20 cases of CPPD-CDD. CPPD-CDD was far more common in women, most frequently involving joints, especially the knees and presenting with various arthrisis. Abnormally calcified and the articular damages were characteristic features by imageing. Histologically, multifocal indigo granular calcinosis was seen in synovium and sometimes appeared as needle-shaped or rhomboid crystals, which characterized the CPPD. Though clinical symptoms of CPPD are quite variable, the definite diagnosis can be made by the abnormal calcification and joint damage radiographically and the indigo CPPD crystals histopathologically.

  8. Identification of monoclinic calcium pyrophosphate dihydrate and hydroxyapatite in human sclera using Raman microspectroscopy

    PubMed Central

    Chen, Ko-Hua; Li, Mei-Jane; Cheng, Wen-Ting; Balic-Zunic, Tonci; Lin, Shan-Yang

    2009-01-01

    Raman microspectroscopy was first used to determine the composition of a calcified plaque located at the pterygium-excision site of a 51-year-old female patient's left nasal sclera after surgery. It was unexpectedly found that the Raman spectrum of the calcified sample at 1149, 1108, 1049, 756, 517, 376 and 352/cm was similar to the Raman spectrum of monoclinic form of calcium pyrophosphate dihydrate (CPPD) crystal, but differed from the Raman spectrum of triclinic form of CPPD. An additional peak at 958/cm was also observed in the Raman spectrum of the calcified plaque, which was identical to the characteristic peak at 958/cm of hydroxyapatite (HA). This is the first study to report the spectral biodiagnosis of both monoclinic CPPD and HA co-deposited in the calcified plaque of a patient with sclera dystrophic calcification using Raman microspectroscopy. PMID:19200254

  9. Calcium pyrophosphate dihydrate deposition disease (CPPD)/Pseudogout of the temporomandibular joint – FNA findings and microanalysis

    PubMed Central

    Naqvi, Asghar H; Abraham, Jerrold L; Kellman, Robert M; Khurana, Kamal K

    2008-01-01

    We report a case of a Calcium pyrophosphate dihydrate deposition disease (CPPD) presenting as a mass in the parotid and temporomandibular joint (TMJ) that simulated a parotid tumor. A 35 year-old man presented with pain in the left ear area. A CT Scan of the area showed a large, calcified mass surrounding the left condylar head, and extending into the infratemporal fossa. FNA of the mass showed birefringent crystals, most of which were rhomboid with occasional ones being needle shaped, embedded in an amorphous pink substance. Scanning electron microscopy (SEM) with energy dispersive x-ray spectroscopy (EDS) of these crystals showed peaks corresponding to calcium and phosphorus. SEM/EDS is a rapid method of diagnosing calcium pyrophosphate dihydrate deposition disease (CPPD) and an alternative to more commonly used method of special staining of cell block sections coupled with polarizing microscopy. PMID:18426573

  10. Enhancement of crystal induced neutrophil responses by opsonisation of calcium pyrophosphate dihydrate crystals.

    PubMed Central

    Burt, H M; Jackson, J K

    1993-01-01

    OBJECTIVES--Little is known about the effect on crystal induced neutrophil responses of the opsonisation of calcium pyrophosphate dihydrate (CPPD) (triclinic) crystals with components of serum and plasma. The purpose of this study was to determine the effects of precoating CPPD crystals with plasma, serum, complement depleted serum, and IgG on a full range of crystal induced neutrophil responses (calcium mobilisation, chemiluminescence, superoxide anion production, non-cytolytic lysosomal enzyme release, and leukotriene synthesis). METHODS--Crystals were precoated with IgG, serum, plasma, or complement depleted serum (heated at 56 degrees C), incubated with neutrophils and the responses monitored with time. Measurement of the extent of neutrophil association with crystals was based on monitoring the decrease in fluorescence intensity of supernatants when crystals and diphenylhexatriene labelled neutrophils were allowed to settle under gravity. RESULTS--Precoating CPPD crystals with IgG, plasma, and serum significantly enhanced chemiluminescence, superoxide anion generation, increases in cytosolic free calcium levels, and non-cytolytic lysosomal enzyme release by neutrophils compared with uncoated CPPD crystals. The enhancement of neutrophil responses by crystals coated with complement depleted serum was less pronounced. The increased neutrophil responses induced by CPPD crystals coated with IgG might have been due to the observed increase in the association of IgG coated crystals with neutrophils. CONCLUSIONS--These data show that there is a marked potentiation of all neutrophil responses to IgG, plasma, and serum coated CPPD crystals. It is suggested that the adsorption of synovial fluid proteins, including IgG and C3b, to CPPD crystals in vivo, results in the opsonised crystals becoming a potent neutrophil stimulant and inflammatory agent. PMID:8215624

  11. The utility of alizarin red s staining in calcium pyrophosphate dihydrate crystal deposition disease.

    PubMed

    Yamakawa, Koji; Iwasaki, Hiroshi; Masuda, Ikuko; Ohjimi, Yuko; Honda, Itsuo; Saeki, Kazuhiko; Zhang, Jingfan; Shono, Eisuke; Naito, Masatoshi; Kikuchi, Masahiro

    2003-05-01

    To determine the most suitable staining method for preservation and detection of calcium pyrophosphate dihydrate (CPPD) crystals in histological sections of patients with CPPD crystal deposition disease. Paraffin sections of CPPD crystal-bearing tissues of 31 patients were stained with hematoxylin and eosin (H&E) and Alizarin red S (ARS). For H&E, the sections were treated with Mayer's hematoxylin (pH 2.3) for 5 min and with eosin alcohol (pH 4.1) for 1 min. For ARS, 1% ARS dissolved in distilled water was adjusted to pH 6.4 by adding 0.1% ammonia solution drop by drop while stirring. As controls, unstained sections were soaked in 1% citric acid monohydrate solution (CAMS, pH 2.3) for 5 or 10 min. The histological preparations were examined under a compensated polarized light using a first-order red compensator. We counted the number of weakly positive birefringent CPPD crystals in 3 high power fields (HPF, 0.272 mm2). CPPD crystals were seen clearly in most specimens stained with ARS, but were markedly reduced in tissue sections stained with H&E or CAMS. The number of CPPD crystals detected in sections stained by ARS (1723 +/- 683 per 3 HPF, mean +/- standard deviation) was significantly higher compared with H&E, CAMS (5 min), and CAMS (10 min) (401 +/- 374, 1022 +/- 616, and 494 +/- 636 per 3 HPF, respectively; p < 0.001, each). Standard H&E staining reduces the number of visible CPPD crystals, probably due to the strong acidity of both hematoxylin and eosin solutions, whereas the ARS stain seems to preserve a large number of CPPD crystals. The utility of ARS staining may improve the identification of CPPD crystals and contribute to a correct diagnosis of CPPD crystal deposition.

  12. Cartilage intermediate layer protein expression in calcium pyrophosphate dihydrate crystal deposition disease.

    PubMed

    Yamakawa, Koji; Iwasaki, Hiroshi; Masuda, Ikuko; Ohjimi, Yuko; Honda, Itsuo; Iyama, Ken-ichi; Shono, Eisuke; Naito, Masatoshi; Kikuchi, Masahiro

    2002-08-01

    To elucidate the mechanisms of calcium pyrophosphate dihydrate crystal deposition disease (CPPDCD) in the meniscus, synovium, labrum, tendon, ligament, and soft tissue, we studied the expression of cartilage intermediate layer protein (CILP). Histological sections and clinical data from 33 patients who fulfilled the criteria of Ryan and McCarty for CPPD were reviewed. Formalin fixed and paraffin embedded tissue sections of 33 patients with CPPDCD were stained with hematoxylin and eosin (H&E) and alizarin red S. Immunostaining was performed using affinity purified polyclonal antibody to synthetic peptide corresponding to the N-terminal sequence of the 61 kDa domain of porcine CILP. The age of patients ranged from 49 to 89 years (median 73). The knee was the commonest site. Radiologically, almost all lesions exhibited fine, radiopaque, linear deposits in the meniscus, articular cartilage, and synovium or joint capsule. Histopathologically, all cases showed deposits of birefringent monoclinic or triclinic crystals, which were visualized by polarized light microscopy with a red analyzer filter. In alizarin red S staining, more numerous crystals were observed than in H&E staining. Crystal deposition was usually associated with adjacent variable amounts of hypertrophic and/or metaplastic chondrocytes in each type of tissue. Variable intensity of CILP immunostaining was found in deposits of each lesion. Hypertrophic/metaplastic chondrocytes in and around CPPD deposits were also positive for CILP. Small cartilaginous islands remote from the CPPD deposits exhibited a weak positivity for CILP. In addition, weakly positive chondrocytes were noted in a transitional zone between cartilaginous islands with and without the deposits. In addition to cytoplasmic immunoreactivity, immunostaining for CILP was observed in the pericellular fibrous matrix. Hypertrophic or metaplastic chondrocytes characteristic of CPPDCD may be directly involved in the formation of CPPD crystals. Our

  13. Preservation of calcium pyrophosphate dihydrate crystals: effect of Mayer's haematoxylin staining period

    PubMed Central

    Ohira, T; Ishikawa, K

    2001-01-01

    OBJECTIVE—To clarify the deleterious effects of Mayer's haematoxylin staining procedure which result in a decrease in, or complete loss of, the number of calcium pyrophosphate dihydrate (CPPD) crystals, and to determine the proper staining period for preserving the crystals in a histological paraffin section of articular tissues.
METHODS—Paraffin sections of CPPD crystal-bearing articular tissues of six patients were stained with Mayer's haematoxylin for 3, 8, or 15 minutes, and subsequently with eosin for one minute. The specimens were examined with an Olympus BHS polarised light microscope. The pH of Mayer's haematoxylin solution was measured with a TOA pH meter.
RESULTS—Positive birefringent CPPD crystals were seen clearly in all specimens stained with Mayer's haematoxylin for three minutes. The specimens stained for eight minutes showed a reduced number of crystals. No crystals were seen in the specimens stained for 15 minutes. Ordinary light microscopy showed no notable differences in the stainability of nucleus, cell membrane, and their surrounding tissues among specimens when stained with Mayer's haematoxylin for either 3, 8, or 15 minutes. The pH of Mayer's haematoxylin solution was 2.31.
CONCLUSIONS—To find CPPD crystals in the paraffin sections of articular tissues, the staining period with Mayer's haematoxylin should be limited to three minutes. The longer the staining period, the greater the reduction in the number of crystals owing to the strong acidity of the haematoxylin solution. A staining period of 15 minutes causes a complete loss of CPPD crystals.

 PMID:11114290

  14. Transmission electron microscopic identification of silicon-containing particles in synovial fluid: potential confusion with calcium pyrophosphate dihydrate and apatite crystals.

    PubMed Central

    Bardin, T; Schumacher, H R; Lansaman, J; Rothfuss, S; Dryll, A

    1984-01-01

    Silicon-containing particles were identified by transmission electron microscopy (TEM) in thin sections of two synovial fluids, which also contained calcium pyrophosphate dihydrate (CPPD) crystals, aspirated during acute attacks of pseudogout. Such particles, which are interpreted as probably being artefacts from glassware, were electron dense and similar in appearance to some CPPD or hydroxyapatite crystals. Images PMID:6476921

  15. The prevalence of chondrocalcinosis of the symphysis pubis on CT scan and correlation with calcium pyrophosphate dihydrate crystal deposition disease.

    PubMed

    Patel, Trusha; Ryan, Lawrence; Dubois, Melissa; Carrera, Guillermo; Baynes, Keith; Mannem, Rajeev; Mulkerin, Jennifer; Visotcky, Alexis

    2016-03-01

    Calcium pyrophosphate dihydrate (CPP) crystal deposition in the articular cartilage can often be seen radiographically as chondrocalcinosis (CC). CPP crystals preferentially deposit in fibrocartilages such as the knee menisci and symphysis pubis (SP). We sought to determine the prevalence of CC in the SP on computed tomography (CT) of the abdomen and pelvis. This retrospective study involved readings on 1070 consecutive CTs of the abdomen and pelvis performed over 3 months in patients over 65 years of age. Medical records of 226 patients found to have CC were reviewed to determine age, gender, documentation of CPPD on problem lists or in medical histories, and whether radiology readings of the CTs mentioned CC. SP CC was identified in 21.1 % (226/1070) of consecutive CT scans with the mean age of CT+ patients being 78.6. Of the 226 patients with SP CC, the observation of CC was documented in only 5.3 % (12/226) of the radiology reports. Of the 12 instances in which the radiology reports mentioned CC, this observation was never (0/12) transmitted to the medical history or problem list. The prevalence of SP CC in patients older than 65 was 21.1 %. Since the majority of CTs of the abdomen and pelvis are not ordered for evaluation of musculoskeletal conditions, this is likely a true prevalence without selection bias. When CC of the SP was present on images, radiologists routinely overlooked or chose not to report CC. Even in the rare instances when it was reported, that information was not added to the medical history or problem list. There are several clinical situations (e.g., acute monoarthritis or atypical osteoarthritis) in which recognizing that a patient has CPP deposition would be useful. Taking the time to review images may yield clinically important findings that are not mentioned anywhere on the patient chart.

  16. Calcium Pyrophosphate Deposition (CPPD)

    MedlinePlus

    ... too. Proper diagnosis depends on detecting calcium pyrophosphate crystals in the fluid of an affected joint. CPPD ... using a microscope to see small calcium pyrophosphate crystals in joint fluid. Anti-inflammatory medications reduce pain ...

  17. Comparison of characteristics of patients with and without calcium pyrophosphate dihydrate crystal deposition disease who underwent total knee replacement surgery for osteoarthritis.

    PubMed

    Viriyavejkul, P; Wilairatana, V; Tanavalee, A; Jaovisidha, K

    2007-02-01

    To compare characteristics of patients with severe osteoarthritis with and without calcium pyrophosphate dihydrate (CPPD) crystal deposition disease. Patients undergoing total knee replacement surgery participated in this study and completed questionnaires. Radiographs of the index knee (extended anteroposterior, lateral and skyline) were reviewed for the presence of chondrocalcinosis. Synovial fluids were obtained during surgery and analyzed under compensated polarized light microscopy. The presence of CPPD crystals was identified in 52.9% of 102 patients. The use of both radiographs and synovial fluid analysis increased the identification of crystals. There was no difference in the following characteristics of the patients with and without CPPD crystals: age of pain onset, gender, difficulty in performing daily functions (including cooking, standing up from chairs, using restroom, going upstairs, and going shopping), history of previous joint inflammation, use of walking aids, and number and types of medications ever used. CPPD patients underwent knee arthroplasty at older age compared to non-CPPD patients (70.3+/-6.37 and 67.5+/-7.15 years old, respectively) (P = 0.037). All but one CPPD patients were unaware of the presence of crystals. High prevalence of CPPD crystals was found in patients undergoing total knee replacement surgery. All but one CPPD patients were unaware of calcium deposition in the index joints. Patients with these crystals experienced similar difficulties in performing daily activities and received similar treatment to patients without CPPD crystals. CPPD patients did not undergo knee arthroplasty at earlier age than non-CPPD patients.

  18. The prevalence of chondrocalcinosis (CC) of the acromioclavicular (AC) joint on chest radiographs and correlation with calcium pyrophosphate dihydrate (CPPD) crystal deposition disease.

    PubMed

    Parperis, Konstantinos; Carrera, Guillermo; Baynes, Keith; Mautz, Alan; Dubois, Melissa; Cerniglia, Ross; Ryan, Lawrence M

    2013-09-01

    Digital imaging combined with picture archiving and communication system (PACS) access allows detailed image retrieval and magnification. Calcium pyrophosphate dihydrate (CPPD) crystals preferentially deposit in fibrocartilages, the cartilage of the acromioclavicular (AC) joint being one such structure. We sought to determine if examination of the AC joints on magnified PACS imaging of chest films would be useful in identifying chondrocalcinosis (CC). Retrospective radiographic readings and chart reviews involving 1,920 patients aged 50 or more who had routine outpatient chest radiographs over a 4-month period were performed. Knee radiographs were available for comparison in 489 patients. Medical records were reviewed to abstract demographics, chest film reports, and diagnoses. AC joint CC was identified in 1.1 % (21/1,920) of consecutive chest films. Patients with AC joint CC were 75 years of age versus 65.4 in those without CC (p < 0.0002). Four hundred eighty-nine patients had knee films. Six of these patients had AC joint CC, and of these, five also had knee CC (83 %). Of the 483 without AC joint CC, 62 (12 %) had knee CC (p = 0.002). Patients with AC joint CC were more likely to have a recorded history of CPPD crystal deposition disease than those without AC joint CC (14 versus 1 %, p = 0.0017). The prevalence of AC joint CC increases with age and is associated with knee CC. A finding of AC joint CC should heighten suspicion of pseudogout or secondary osteoarthritis in appropriate clinical settings and, in a young patient, should alert the clinician to the possibility of an associated metabolic condition.

  19. Activation of S6 kinase in human neutrophils by calcium pyrophosphate dihydrate crystals: protein kinase C-dependent and phosphatidylinositol-3-kinase-independent pathways.

    PubMed Central

    Tudan, C; Jackson, J K; Charlton, L; Pelech, S L; Sahl, B; Burt, H M

    1998-01-01

    Phosphatidylinositol 3-kinase (PI 3-kinase) has been shown previously to be a central enzyme in crystal-induced neutrophil activation. Since activation of the 70 kDa S6 kinase (p70S6K) has been shown to be dependent on PI 3-kinase activation in mammalian cells, and since the former is a key enzyme in the transmission of signals to the cell nucleus, activation of p70(S6K) was investigated in crystal-stimulated neutrophils. Cytosolic fractions from calcium pyrophosphate dihydrate (CPPD)-crystal-activated neutrophils were separated by Mono Q chromatography and analysed for phosphotransferase activity using a range of substrates and probed by Western analysis using antibodies to p70(S6K) and mitogen-activated protein kinase (MAP kinase). CPPD crystals induced a robust, transient activation (peak activity at 2 min) of p70(S6K) that was fully inhibited by pretreatment with rapamycin. This is the first report of the activation of p70(S6K) in neutrophil signal transduction pathways induced by an agonist. This crystal-induced activation of p70(S6K) could also be inhibited by a protein kinase C (PKC) inhibitor (Compound 3), but not by the PI 3-kinase inhibitor wortmannin. CPPD crystals also activated the ERK1 and ERK2 forms of MAP kinase (wortmannin insensitive), PKC (Compound 3 sensitive) and protein kinase B (wortmannin sensitive) in neutrophils. These data suggest that activation of p70(S6K) may proceed through a PI 3-kinase- and protein kinase B-independent but PKC-dependent pathway in crystal-activated neutrophils. PMID:9531494

  20. Calcium pyrophosphate dihydrate crystals activate MAP kinase in human neutrophils: inhibition of MAP kinase, oxidase activation and degranulation responses of neutrophils by taxol.

    PubMed Central

    Jackson, J K; Tudan, C; Sahl, B; Pelech, S L; Burt, H M

    1997-01-01

    The activation of MAP kinase in human neutrophils stimulated by both uncoated and plasma-opsonized crystals of triclinic calcium pyrophosphate dihydrate (CPPD) was investigated. The effect of taxol on MAP kinase activation and on the responses of neutrophils stimulated by plasma-opsonized crystals was determined. MAP kinase activation was identified and quantified in Mono Q chromatography separated fractions of neutrophils that had been incubated with CPPD crystals by measuring [gamma-32P]adenosine triphosphate (ATP) phosphorylation of myelin basic protein and using immunoblotting techniques. Human neutrophils were incubated with taxol (0-50 microM), added to plasma-opsonized CPPD (50 mg/ml) and MAP kinase activation, chemiluminescence, superoxide anion generation, lysozyme and myeloperoxidase release were monitored. Both uncoated and plasma coated CPPD crystals induced a large increase in MAP kinase activity in neutrophils over control levels within 1 min of incubation. Pretreatment of neutrophils with taxol was able to suppress this activation of MAP kinase. Taxol produced a concentration-dependent inhibition of opsonized CPPD-induced neutrophil chemiluminescence, superoxide anion production and myeloperoxide release. Taxol at 28 microM also significantly inhibited chemiluminescence, superoxide anion production and myeloperoxidase release from neutrophils stimulated by opsonized zymosan. This is the first report of crystal-induced activation of MAP kinase in neutrophils. Microtubule-associated processes, such as signal transduction, secretion and phagocytosis are involved in particulate-induced neutrophil responses. We have suggested that the inhibitory effect of taxol observed in this work is due to its stabilizing effect on microtubules and disruption of MAP kinase activation associated with microtubules. Images Figure 1 Figure 3 PMID:9176102

  1. Up-regulated expression of cartilage intermediate-layer protein and ANK in articular hyaline cartilage from patients with calcium pyrophosphate dihydrate crystal deposition disease.

    PubMed

    Hirose, Jun; Ryan, Lawrence M; Masuda, Ikuko

    2002-12-01

    Excess accumulation of extracellular inorganic pyrophosphate (ePPi) in aged human cartilage is crucial in calcium pyrophosphate dihydrate (CPPD) crystal formation in cartilage matrix. Two sources of ePPi are ePPi-generating ectoenzymes (NTPPPH) and extracellular transport of intracellular PPi by ANK. This study was undertaken to evaluate the role of NTPPPH and ANK in ePPi elaboration, by investigating expression of NTPPPH enzymes (cartilage intermediate-layer protein [CILP] and plasma cell membrane glycoprotein 1 [PC-1]) and ANK in human chondrocytes from osteoarthritic (OA) articular cartilage containing CPPD crystals and without crystals. Chondrocytes were harvested from knee cartilage at the time of arthroplasty (OA with CPPD crystals [CPPD], n = 8; OA without crystals [OA], n = 10). Normal adult human chondrocytes (n = 1) were used as a control. Chondrocytes were cultured with transforming growth factor beta1 (TGFbeta1), which stimulates ePPi elaboration, and/or insulin-like growth factor 1 (IGF-1), which inhibits ePPi elaboration. NTPPPH and ePPi were measured in the media at 48 hours. Media CILP, PC-1, and ANK were determined by dot-immunoblot analysis. Chondrocyte messenger RNA (mRNA) was extracted for reverse transcriptase-polymerase chain reaction to study expression of mRNA for CILP, PC-1, and ANK. NTPPPH and ANK mRNA and protein were also studied in fresh frozen cartilage. Basal ePPi elaboration and NTPPPH activity in conditioned media from CPPD chondrocytes were elevated compared with normal chondrocytes, and tended to be higher compared with OA chondrocytes. Basal expression of mRNA for CILP (chondrocytes) and ANK (cartilage) was higher in both CPPD chondrocytes and CPPD cartilage extract than in OA or normal samples. PC-1 mRNA was less abundant in CPPD chondrocytes and cartilage extract than in OA chondrocytes and extract, although the difference was not significant. CILP, PC-1, and ANK protein levels were similar in CPPD, OA, and normal chondrocytes

  2. 21 CFR 582.5223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Calcium pyrophosphate. 582.5223 Section 582.5223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5223 Calcium pyrophosphate. (a) Product. Calcium pyrophosphate. (b) Conditions of use...

  3. 21 CFR 582.5223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Calcium pyrophosphate. 582.5223 Section 582.5223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5223 Calcium pyrophosphate. (a) Product. Calcium pyrophosphate. (b) Conditions of use...

  4. 21 CFR 582.5223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Calcium pyrophosphate. 582.5223 Section 582.5223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5223 Calcium pyrophosphate. (a) Product. Calcium pyrophosphate. (b) Conditions of use...

  5. 21 CFR 582.5223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Calcium pyrophosphate. 582.5223 Section 582.5223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5223 Calcium pyrophosphate. (a) Product. Calcium pyrophosphate. (b) Conditions of use...

  6. Acute calcium pyrophosphate deposition arthropathy.

    PubMed

    Rosen, Thomas; Furman, Janet

    2016-06-01

    Acute calcium pyrophosphate deposition (CPPD) arthropathy, also called pseudogout, is common, and becomes more prevalent as patients age. The presenting symptoms are similar to both gout and septic arthritis but may be treated differently. This article describes a typical patient presentation and management from an emergency medicine and orthopedic surgery standpoint.

  7. 21 CFR 182.8223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Calcium pyrophosphate. 182.8223 Section 182.8223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients § 182.8223 Calcium pyrophosphate. (a) Product....

  8. Synthesis of triclinic calcium pyrophosphate crystals.

    PubMed

    Groves, P J; Wilson, R M; Dieppe, P A; Shellis, R P

    2007-07-01

    This paper presents a method for preparing crystals of triclinic calcium pyrophosphate (t-CPPD). A calcium pyrophosphate intermediate is first prepared by reaction of potassium pyrophosphate and calcium chloride. Samples of the intermediate are dissolved in hydrochloric acid and urea added. Upon heating to 95-100 degrees C, hydrolysis of the urea causes the pH to rise and t-CPPD crystallises out. Purity of the product was ascertained by chemical and physical analysis. Where large crystals are required an unstirred system is used, while smaller crystals are produced by stirring the reaction mixture.

  9. Bilateral Olecranon Bursitis – A Rare Clinical presentation of Calcium Pyrophosphate Crystal Deposition Disease

    PubMed Central

    Patel, Jignesh; Girishkumar; Mruthyunjaya; Rupakumar, C. S

    2014-01-01

    Introduction: Calcium pyrophosphate crystal deposition disease (CPPD) is the most common form of crystal arthropathy second only to gout. Common clinical presentation is an acute monoarticular arthritis commonly occurring in knee joints. We presented a case of bilateral olecranon bursitis in a calcium pyrophosphate crystal deposition disease. Case Report: A 42-year-old female patient is presented with golf ball sized painless swellings in the posterior aspect of her elbows. Elbow joints were clinically normal except for restriction of terminal flexion. X-ray showed mild erosion at the tip of olecranon. Excision biopsy of the swelling showed positive birefringent calcium pyrophosphate dehydrate crystals on the inner wall of the specimen on polarized light microscopy. Conclusion: Bilateral olecranon bursitis may be part of the extraarticular manifestations of calcium pyrophosphate dihydrate crystal deposition disease with good prognosis following in toto bursa excision. PMID:27298934

  10. 21 CFR 182.8223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium pyrophosphate. 182.8223 Section 182.8223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients § 182.8223 Calcium...

  11. 21 CFR 182.8223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Calcium pyrophosphate. 182.8223 Section 182.8223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients § 182.8223 Calcium...

  12. Synthesis and Characterization of Nanoparticles of Calcium Pyrophosphate

    NASA Astrophysics Data System (ADS)

    Vasant, Sonal R.; Joshi, M. J.

    Calcium phosphate based biomaterials play important roles in clinical applications. Calcium pyrophosphate (CPP), a kind of calcium phosphate, can be used as a bone substitution material as well as a bone graft. Because of its similarity to inorganic component of bone and teeth it can be used for surface coating of metallic dental and orthopedic implants. In the present study, calcium pyrophosphate dihydrate (CPPD) nanoparticles were synthesized using surfactant mediated approach. Crystalline nature and average crystallite size was studied using Powder XRD. The CPPD nanocrystallites were found to be triclinic from powder XRD. The TEM study indicated that CPPD nanoparticles were in the range of 13 nm to 20 nm. The presence of various bonds was confirmed by FTIR spectroscopy. The amount of water of hydration and the thermal stability was studied by thermogravimetry. The variations of various dielectric parameters with the frequency of applied field in 3.2 kHz to 32 MHz range and within a temperature range from 60°C to 120°C were studied. The formation of other phases such as β-CPP and α-CPP on heating of CPPD at 900°C and 1250°C, respectively, were studied by the Powder XRD. The results are discussed.

  13. 21 CFR 182.8223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Calcium pyrophosphate. 182.8223 Section 182.8223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients § 182.8223...

  14. 21 CFR 582.5223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Calcium pyrophosphate. 582.5223 Section 582.5223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or...

  15. 21 CFR 182.8223 - Calcium pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Calcium pyrophosphate. 182.8223 Section 182.8223 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients § 182.8223...

  16. Structure of the calcium pyrophosphate monohydrate phase (Ca2P2O7·H2O): towards understanding the dehydration process in calcium pyrophosphate hydrates.

    PubMed

    Gras, Pierre; Ratel-Ramond, Nicolas; Teychéné, Sébastien; Rey, Christian; Elkaim, Erik; Biscans, Béatrice; Sarda, Stéphanie; Combes, Christèle

    2014-09-01

    Calcium pyrophosphate hydrate (CPP, Ca(2)P(2)O(7) · nH2O) and calcium orthophosphate compounds (including apatite, octacalcium phosphate etc.) are among the most prevalent pathological calcifications in joints. Even though only two dihydrated forms of CPP (CPPD) have been detected in vivo (monoclinic and triclinic CPPD), investigations of other hydrated forms such as tetrahydrated or amorphous CPP are relevant to a further understanding of the physicochemistry of those phases of biological interest. The synthesis of single crystals of calcium pyrophosphate monohydrate (CPPM; Ca(2)P(2)O(7) · H2O) by diffusion in silica gel at ambient temperature and the structural analysis of this phase are reported in this paper. Complementarily, data from synchrotron X-ray diffraction on a CPPM powder sample have been fitted to the crystal parameters. Finally, the relationship between the resolved structure for the CPPM phase and the structure of the tetrahydrated calcium pyrophosphate β phase (CPPT-β) is discussed.

  17. Two hydrate pseudopolymorphs of thiamine pyrophosphate: a dihydrate and a trihydrate.

    PubMed

    Li, Shu-Qiang; Hu, Ning-Hai

    2013-07-01

    Two hydrate pseudopolymorphs of 3-[(4-amino-2-methylpyrimidin-1-ium-5-yl)methyl]-4-methyl-1,3-thiazol-3-ium-5-yl hydrogen pyrophosphate (TPP), viz. a dihydrate, C12H18N4O7P2S·2H2O, (I), and a trihydrate, C12H18N4O7P2S·3H2O, (II), were obtained during a structural study of vitamin B1 coenzyme. In both compounds, TPP is a neutral zwitterion, with its pyrophosphate group doubly deprotonated and its pyrimidine ring protonated, and it assumes the usual `F' conformation in terms of the two torsion angles about the bonds by which the methylene group links the thiazolium and pyrimidinium rings [1.1 (3) and 79.7 (3)° for (I), and 2.0 (3) and 75.5 (3)° for (II)]. In (I), two TPP molecules are linked by a pair of O-H···O hydrogen bonds into a phosphate-pairing dimer. N-H···O hydrogen bonds connect the dimers into a sheet parallel to (101). In (II), the TPP molecules are self-assembled solely by N-H···O hydrogen bonds, generating a tape structure along [001]. A comparison of the four known hydrate pseudopolymorphs of TPP shows that the phosphate-pairing dimers are basic building units for the formation of two-dimensional networks.

  18. Titanium implant osseointegration with calcium pyrophosphate in rabbits.

    PubMed

    Koo, Samuel; König, Bruno; Allegrini, Sérgio; Yoshimoto, Marcelo; Carbonari, Marcelo José; Mitri-Luiz, Fábio Franceschini

    2006-02-01

    The objective of this study was to characterize calcium pyrophosphate material, evaluate its in vitro cytotoxicity, and assess its ability to induce bone formation. X-ray diffraction (XRD) was used to determine crystallinity and phases present in material. Serial dilutions of extracts, from 10-day dissolution tests in modified Eagle's medium, were exposed for 24 h to mouse fibroblasts and cytotoxicity assessed via viable staining. In vivo performance was determined by placing Ti screws with and without calcium pyrophosphate agglutinated with marrow adipose tissue in the tibiae of eight rabbits. New bone formation around test and control implants was evaluated histomorphometrically by using three fluorochrome labels: alizarin, calcein, and tetracycline. After 8 postoperative weeks, the animals were killed and specimens were retrieved and processed for fluorescence and light microscopic analysis. Calcium pyrophosphate showed no cytotoxicity and the XRD showed that the main phase of the analyzed sample corresponded to beta-calcium pyrophosphate. The largest fluorochrome labeling area occurred during the fourth and fifth postoperative weeks, in both control and experimental groups. Histologically, the bone neoformation occurred in regions where the calcium pyrophosphate was resorbed. The morphometric analysis showed implants placed with calcium pyrophosphate resulted in smaller polyfluorochrome labeling area (p < 0.05). Copyright 2005 Wiley Periodicals, Inc.

  19. Hyaline cartilage involvement in patients with gout and calcium pyrophosphate deposition disease. An ultrasound study.

    PubMed

    Filippucci, E; Riveros, M Gutierrez; Georgescu, D; Salaffi, F; Grassi, W

    2009-02-01

    The main aim of the present study was to determine the sensitivity, specificity and accuracy of ultrasonography (US) in detecting monosodium urate and calcium pyrophosphate dihydrate crystals deposits at knee cartilage level using clinical definite diagnosis as standard reference. A total of 32 patients with a diagnosis of gout and 48 patients with pyrophosphate arthropathy were included in the study. Fifty-two patients with rheumatoid arthritis (RA), psoriatic arthritis or osteoarthritis (OA) were recruited as disease controls. All diagnoses were made using an international clinical criterion. US examinations were performed by an experienced sonographer, blind to clinical and laboratory data. Hyaline cartilage was assessed to detect two US findings recently indicated as indicative of crystal deposits: hyperechoic enhancement of the superficial margin of the hyaline cartilage and hyperechoic spots within the cartilage layer not generating a posterior acoustic shadow. Hyperechoic enhancement of the chondrosynovial margin was found in at least one knee of 14 out of 32 (43.7%) patients with gout and in a single knee of only one patient affected by pyrophosphate arthropathy (specificity=99%). Intra-cartilaginous hyperechoic spots were detected in at least one knee of 33 out of 48 (68.7%) patients with pyrophosphate arthropathy and in two disease controls one with OA and the second with RA (specificity=97.6%). The results of the present study indicate that US may play a relevant role in distinguishing cartilage involvement in patients with crystal-related arthropathy. The selected US findings were found to be highly specific.

  20. Most calcium pyrophosphate crystals appear as non-birefringent

    PubMed Central

    Ivorra, J.; Rosas, J.; Pascual, E.

    1999-01-01

    OBJECTIVE—To determine the proportion of calcium pyrophosphate dihydrate (CPPD) crystals that appear as non-birefringent when observed under the polarised light microscope.
METHODS—Two observers examined independently 10 synovial fluid samples obtained during an episode of arthritis attributable to CPPD crystals. Ten synovial fluid samples from patients with acute gout were used as a reference. The examination was performed after placing a fluid sample in a Niebauer haemocytometric chamber; a crystal count was done first under ordinary light, then in the area corresponding to a 0.1 ml, under polarised light
RESULTS—The percentages of birefringence appreciated for CPPD were 18% (confidence intervals (CI) 12, 24) for observer 1, and 17% (CI 10, 24) for observer 2 (difference NS). The percentages of birefringence for monosodium urate were 127% (CI 103, 151) for observer 1 and 107% (CI 100, 114) for observer 2 (difference NS). Percentages above 100% indicate that crystals missed under ordinary light became apparent under polarised light.
CONCLUSION—Only about one fifth of all CPPD crystals identified by bright field microscopy show birefringence when the same synovial fluid sample is observed under polarised light. If a search for CPPD crystals is conducted under polarised light, the majority of the crystals will be missed. Ordinary light allows a better rate of CPPD crystal detection but observation under polarised light of crystals showing birefringence is required for definitive CPPD crystal identification.

 PMID:10460193

  1. Hand and wrist arthropathies of hemochromatosis and calcium pyrophosphate deposition disease: distinct radiographic features

    SciTech Connect

    Adamson, T.C. III; Resnik, C.S.; Guerra, J. Jr.; Vint, V.C.; Weisman, M.H.; Resnick, D.

    1983-05-01

    Radiographic features of hand and wrist involvement in 26 patients with hemochromatosis and in 26 patients with idiopathic calcium pyrophosphate dihydrate (CPPD) crystal deposition disease were compared. Two radiologists independently examined the radiographs without knowledge of the specific group to which the patient belonged. The results of this study clearly establish that structural joint diseases in the two disorders are not identical. Characteristic findings allow the radiologist to favor one diagnosis over the other. These radiograhic differences indicate that the arthropathy of hemochromatosis is related to factors additional to the presence of CPPD crystals, specifically, the more prevalent narrowing of the metacarpophalangeal joint spaces, including those in the fourth and fifth digits, peculiar hook-like osteophytes on the radial aspect of the metacarpal heads, and less prevalent separation of the scaphoid and the lunate.

  2. Powder XRD and dielectric studies of gel grown calcium pyrophosphate crystals

    NASA Astrophysics Data System (ADS)

    Parekh, Bharat; Parikh, Ketan; Joshi, Mihir

    2013-06-01

    Formation of calcium pyrophosphate dihydrate (CPPD) crystals in soft tissues such as cartilage, meniscus and synovial tissue leads to CPPD deposition diseases. The appearance of these crystals in the synovial fluid can give rise to an acute arthritic attack with pain and inflammation of the joints, a condition called pseudo-gout. The growth of CPP crystals has been carried out, in the present study, using the single diffusion gel growth technique, which can broadly mimic in vitro the condition in soft tissues. The crystals were characterized by different techniques. The FTIR study revealed the presence of various functional groups. Powder XRD study was also carried out to verify the crystal structure. The dielectric study was carried out at room temperature by applying field of different frequency from 500 Hz to 1 MHz. The dielectric constant, dielectric loss and a.c. resistivity decreased as frequency increased, whereas the a.c. conductivity increased as frequency increased.

  3. Preparation and characterization of RF magnetron sputtered calcium pyrophosphate coatings.

    PubMed

    Yonggang, Yan; Wolke, J G C; Yubao, Li; Jansen, J A

    2006-03-15

    CaP ceramic has been widely used as coating on metals in orthopedics and oral dentistry. Variations in CaP composition can lead to different dissolution/precipitation behavior and may also affect the bone response. In the present study calcium pyrophosphate and hydroxylapatite coatings were successfully prepared by RF magnetron sputtering deposition. The phase composition, morphological properties, and the dissolution in SBF were characterized by using XRD, FTIR, EDS, SEM, and spectrophotometry. The results showed that all the sputtered coatings were amorphous and changed into a crystal structure after IR-radiation. The temperature for the crystallization of the amorphous coatings is lower for the hydroxylapatite coating (550 degrees C), compared to the calcium pyrophosphate coating (650 degrees C). All sputtered amorphous coatings were instable in SBF and dissolved partially within 4 wks of incubation. The heat-treated coatings appeared to be stable after incubation. These results showed that magnetron sputtering of calcium pyrophosphate coating is a promising method for forming a biocompatible ceramic coating.

  4. A new precipitation pathway for calcium sulfate dihydrate (gypsum) via amorphous and hemihydrate intermediates.

    PubMed

    Wang, Yun-Wei; Kim, Yi-Yeoun; Christenson, Hugo K; Meldrum, Fiona C

    2012-01-14

    This work investigates the early stages of precipitation of calcium sulfate from aqueous solution at room temperature and shows for the first time that amorphous calcium sulfate (ACS) and calcium sulfate hemihydrate are sequentially precipitated prior to calcium sulfate dihydrate (gypsum). This journal is © The Royal Society of Chemistry 2012

  5. Dibasic calcium phosphate dihydrate, USP material compatibility with gamma radiation

    NASA Astrophysics Data System (ADS)

    Betancourt Quiles, Maritza

    Gamma radiation is a commonly used method to reduce the microbial bioburden in compatible materials when it is applied at appropriate dose levels. Gamma irradiation kills bacteria and mold by breaking down the organism’s DNA and inhibiting cell division. The purpose of this study is to determine the radiation dosage to be used to treat Dibasic Calcium Phosphate Dihydrate, USP (DCPD) and to evaluate its physicochemical effects if any, on this material. This material will be submitted to various doses of gamma radiation that were selected based on literature review and existing regulations that demonstrate that this method is effective to reduce or eliminate microbial bioburden in natural source and synthetic materials. Analytical testing was conducted to the DCPD exposed material in order to demonstrate that gamma radiation does not alter the physicochemical properties and material still acceptable for use in the manufacture of pharmaceutical products. The results obtained through this study were satisfactory and demonstrated that the gamma irradiation dosages from 5 to 30 kGy can be applied to DCPD without altering its physicochemical properties. These are supported by the Assay test data evaluation of lots tested before and after gamma irradiation implementation that show no significant statistical difference between irradiated and non irradiated assay results. The results of this study represent an achievement for the industry since they provide as an alternative the use of Gamma irradiation technology to control the microbial growth in DCPD.

  6. Calcium sulfate dihydrate urolithiasis in a pet rabbit.

    PubMed

    Kucera, Jaroslav; Koristkova, Tamara; Gottwaldova, Barbora; Jekl, Vladimir

    2017-03-01

    CASE DESCRIPTION A 3-year-old sexually intact male rabbit (Oryctolagus cuniculus) was evaluated because of a 1-day history of signs of anorexia and depression. CLINICAL FINDINGS Clinical examination revealed signs of depression, hunched posture, low skin elasticity (suggesting dehydration), slightly distended abdomen, and penile and preputial edema. The owner reported that the rabbit had been fed a routine diet, received water via a sipper bottle, and was allowed free movement around the home. It had been observed by the owner to bite and chew gypsum-based plaster from the walls of the home. Abdominal radiography and ultrasonography revealed radiopaque material in the urinary bladder, irregular thickening of the urinary bladder wall, and gaseous distention of the cecum. Urinalysis revealed mild hematuria and proteinuria. Results of the physical examination and other diagnostic tests were consistent with urolithiasis, cystitis, and gastrointestinal stasis. TREATMENT AND OUTCOME At clinical examination, numerous small uroliths originating from the urethral orifice were removed and submitted for composition analysis via infrared and Raman spectrometry and polarized microscopy. Laparotomy-assisted flushing of the urinary bladder and urethra was performed, and the rabbit recovered without complication. Results of composition analysis indicated the uroliths were composed of calcium sulfate dihydrate. CLINICAL RELEVANCE This is the first report of calcium sulfate urolithiasis in a rabbit, which was attributed to dehydration (possibly due to inadequate water provision) and excessive dietary intake of sulfur in the form of gypsum-based plaster. Rabbits should be prevented from consuming plaster and other potential extradietary sources of sulfur and provided an appropriate water supply.

  7. Change perspective to increase diagnostic accuracy of ultrasonography in calcium pyrophosphate dehydrate deposition disease! A new approach: the axial scan of the meniscus.

    PubMed

    Filippou, G; Picerno, V; Adinolfi, A; Di Sabatino, V; Bertoldi, I; Galeazzi, M; Frediani, B

    2015-03-31

    Ultrasonography (US) is a relevant tool in the study of calcium pyrophosphate dihydrate (CPP) deposition disease. However, differential diagnosis of hyperechoic deposits within the fibrocartilage can be difficult; moreover, US study is limited by the need of an adequate acoustic window. We describe a US scanning technique that offers a new viewpoint in the study of knee meniscal structure: a longitudinal scan performed according to the long axis of meniscus. This technique proves to be particularly useful for the identification of CPP deposition, but could also improve the US diagnostic utility and accuracy in other meniscal pathologies.

  8. Tri-calcium phosphate (ß-TCP) can be artificially synthesized by recycling dihydrate gypsum hardened.

    PubMed

    Han-Cheol, Cho; Hori, Masaharu; Yoshida, Takakazu; Yamada, Naoko; Komada, Yuko; Tamaki, Yukimichi; Miyazaki, Takashi

    2014-01-01

    Calcium phosphate is known as a major component of biological hard tissues. This study aimed to produce calcium phosphate by recycling kneaded surplus gypsum. β-dihydrate gypsum was derived from commercial dental β-hemihydrate gypsum, which was mechanically powdered and mixed with the liquid component of a commercial zinc phosphate cement. This mixture was fired at 1,200°C and evaluated by XRD analysis, thermal analysis and scanning electron microscopy (SEM). An acceptable ratio of mixing was 4 g of β-dihydrate gypsum powder to 1.5 mL of phosphoric acid liquid. XRD peaks were monotonic below 800°C, but new ß-TCP was formed by firing at 900°C or more, although TG-DTA analysis of synthetic ß-TCP suggested that some residual dihydrate gypsum remained in the sample. SEM images indicated a fused-block bone-like structure covered with phosphorus and calcium. These results suggest that production of synthetic β-TCP is possible through ecological techniques using recycled materials.

  9. Effects of pyrophosphate ions on protein adsorption onto calcium hydroxyapatite.

    PubMed

    Kandori, Kazuhiko; Oda, Shohei; Tsuyama, Shintaro

    2008-02-28

    The effects of pyrophosphate ions (PP: P2O7(4-)) on the adsorption of proteins onto calcium hydroxyapatite (Hap) were examined using typical proteins of bovine serum albumin (BSA: isoelectric point (iep) = 4.7, molecular mass (M(s)) = 67 200 Da, acidic protein), myoglobin (MGB: iep = 7.0, M(s) = 17 800 Da, neutral protein), and lysozyme (LSZ: iep = 11.1, M(s) = 14,600 Da, basic protein). The UV and CD measurements determined that both the secondary and the tertiary structures of protein molecules do not vary in the presence of PP. The adsorption of BSA was strongly depressed by the addition of PP in all the methods with changing the order of PP addition. Even if BSA was pre-adsorbed on the Hap surface, PP replaced BSA molecules by strong preferential adsorption onto Hap to reduce the amounts of adsorbed BSA. A similar effect was observed with the adsorption of MGB. On the other hand, the amount of adsorbed LSZ (n(LSZ)) was increased with an increase in the concentration of PP, and the n(LSZ) value showed a maximum point in each adsorption isotherm. This fact was explained by a compression of the electric double layer (EDL) around each LSZ molecule by PP. This compression of the EDL induced the reduction of lateral electrostatic repulsions between charged LSZ molecules on the Hap surface and enhanced the formation of closed-packed monolayers to raise the n(LSZ) value. However, since the number of PPs around a LSZ molecule is decreased by an increase in the LSZ concentration in each system, the thickness of the EDL may be increased. Hence, n(LSZ) was reduced again after the maximum point in each system. Tripolyphosphate (TPP: P3O10(5-)) ions exhibited similar effects on the adsorption behaviors of all proteins, but a much more pronounced effect was observed on the LSZ system. TPP with a higher eletronegativity shielded the EDL more highly than PP to increase the n(LSZ) value. The results of the zeta potential for all the protein systems supported the modes of protein

  10. Proteomic analysis of a rare urinary stone composed of calcium carbonate and calcium oxalate dihydrate: a case report.

    PubMed

    Kaneko, Kiyoko; Matsuta, Yosuke; Moriyama, Manabu; Yasuda, Makoto; Chishima, Noriharu; Yamaoka, Noriko; Fukuuchi, Tomoko; Miyazawa, Katsuhito; Suzuki, Koji

    2014-03-01

    The objective of the present study was to investigate the matrix protein of a rare urinary stone that contained calcium carbonate. A urinary stone was extracted from a 34-year-old male patient with metabolic alkalosis. After X-ray diffractometry and infrared analysis of the stone, proteomic analysis was carried out. The resulting mass spectra were evaluated with protein search software, and matrix proteins were identified. X-ray diffraction and infrared analysis confirmed that the stone contained calcium carbonate and calcium oxalate dihydrate. Of the identified 53 proteins, 24 have not been previously reported from calcium oxalate- or calcium phosphate-containing stones. The protease inhibitors and several proteins related to cell adhesion or the cytoskeleton were identified for the first time. We analyzed in detail a rare urinary stone composed of calcium carbonate and calcium oxalate dihydrate. Considering the formation of a calcium carbonate stone, the new identified proteins should play an important role on the urolithiasis process in alkaline condition. © 2013 The Japanese Urological Association.

  11. Determination of dosimetric and kinetic features of gamma irradiated solid calcium ascorbate dihydrate using ESR spectroscopy

    NASA Astrophysics Data System (ADS)

    Tuner, H.

    2013-01-01

    Effects of gamma radiation on solid calcium ascorbate dihydrate were studied using electron spin resonance (ESR) spectroscopy. Irradiated samples were found to present two specific ESR lines with shoulder at low and high magnetic field sides. Structural and kinetic features of the radicalic species responsible for experimental ESR spectrum were explored through the variations of the signal intensities with applied microwave power, variable temperature, high-temperature annealing and room temperature storage time studies. Dosimetric potential of the sample was also determined using spectrum area and measured signal intensity measurements. It was concluded that three radicals with different spectroscopic and kinetic features were produced upon gamma irradiation.

  12. Unusual effect of water vapor pressure on dehydration of dibasic calcium phosphate dihydrate.

    PubMed

    Kaushal, Aditya M; Vangala, Venu R; Suryanarayanan, Raj

    2011-04-01

    Dibasic calcium phosphate occurs as an anhydrate (DCPA; CaHPO₄) and as a dihydrate (DCPD; CaHPO₄•2H₂O). Our objective was to investigate the unusual behavior of these phases. Dibasic calcium phosphate dihydrate was dehydrated in a (i) differential scanning calorimeter (DSC) in different pan configurations; (ii) variable-temperature X-ray diffractometer (XRD) at atmospheric and under reduced pressure, and in sealed capillaries; and (iii) water vapor sorption analyzer at varying temperature and humidity conditions. Dehydration was complete by 210°C in an open DSC pan and under atmospheric pressure in the XRD. Unlike "conventional" hydrates, the dehydration of DCPD was facilitated in the presence of water vapor. Variable-temperature XRD in a sealed capillary and DSC in a hermetic pan with pinhole caused complete dehydration by 100°C and 140°C, respectively. Under reduced pressure, conversion to the anhydrate was incomplete even at 300°C. The increase in dehydration rate with increase in water vapor pressure has been explained by the Smith-Topley effect. Under "dry" conditions, a coating of poorly crystalline product is believed to form on the surface of particles and act as a barrier to further dehydration. However, in the presence of water vapor, recrystallization occurs, creating cracks and channels and facilitating continued dehydration.

  13. Cytotoxicity and migration of fibroblasts on two types of calcium sulfate dihydrate.

    PubMed

    Teparat-Burana, Thitiwan; Onsiri, Natnicha; Jantarat, Jeeraphat

    2017-02-01

    The aim of the present study was to compare cytotoxicity and migration ability of L929 on medical-grade calcium sulfate dihydrate (MCS) with commercial calcium sulfate dihydrate (CCS). Samples of both freshly-mixed and set states of MCS and CCS were extracted in culture medium and tested for cytotoxicity according to International Organization of Standardization 10993-5:2009. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used, and the percentage of cell viability was calculated. Thirty petri dishes were divided into three equal groups: polystyrene (control), CCS, and MCS. In each petri dish, triplicate wells were made on the agarose overlay on the material. The central well was occupied with L929 cell suspension, one well was filled with platelet-derived growth factor-BB, and the remaining well with Hanks' Balanced Salt Solution. After incubation, the migration distances were measured and the corrected migration distances were calculated. The final step involved scanning electron microscopy (SEM). The CCS had significantly less cell viability than MCS at a high extracted concentration (P < 0.001) in both samples. The mean corrected migration of MCS was significantly greater than that of CCS (P < 0.05). SEM showed that L929 cells on MCS exhibited an elongated spindle shape. MCS was less cytotoxic and provided greater migration of L929 fibroblasts compared with CCS. © 2015 Wiley Publishing Asia Pty Ltd.

  14. Bone kinetics of calcium-45 and pyrophosphate labeled with technetium-96: an autoradiographic evaluation. [Rabbits

    SciTech Connect

    Guillemart, A.; Le Pape, A.; Galy, G.; Besnard, J.C.

    1980-05-01

    The uptake of calcium-45 and of pyrophosphate labeled with the long-lived technetium-96 isotope was compared by means of liquid-emulsion microautoradiograms of the epiphyseal plates of 10-week-old rabbits, at 30 min, and 3 and 48 hr after i.v administration. For both tracers, thin sections confirm the significant role of the blood supply, especially shortly after injection. However, other more specific mechanisms lead to a mixing of the calcium in the mineral mass and to a linear deposition of technetium facing the osteoid surfaces. These findings suggest that the tropism of tin-reduced technetium pyrophosphate is not governed by the mineral pool but rather by exchanges inside a still poorly calcified organic matrix.

  15. Crystal arthritides - gout and calcium pyrophosphate arthritis : Part 1: Epidemiology and pathophysiology.

    PubMed

    Schlee, S; Bollheimer, L C; Bertsch, T; Sieber, C C; Härle, P

    2017-02-23

    Gout and calcium pyrophosphate deposition disease (CPPD, pseudogout) are still the most frequent inflammatory arthritides in multimorbid elderly patients. Gout and CPPD are different diseases and based on different pathophysiological principles. Gout is closely associated with the metabolic syndrome and is an independent risk factor for cardiovascular mortality. The prevalence of asymptomatic hyperuricemia is estimated to be 10-20% of adults in industrial nations and prevalence is strongly associated with age. More than 7% of persons aged over 65 years suffer from clinically manifest gout. The underlying pathophysiological principle is an imbalance between the formation and elimination of uric acid. The degradation of the purine bases adenine and guanosine to uric acid is catalysed by xanthine oxidase and genetic polymorphisms and mutations play an important role in absorption and excretion processes. Furthermore, carrier proteins, such as URAT-1 or OAT-4 also have an influence on these processes. An imbalance of the physiological processes results in the solubility product being exceeded, which in consequence leads to crystallization of urate. This induces a cascade of massive inflammatory reactions at the molecular and cellular level with the activation of cytokines. The inflammatory process can be stopped by neutrophil extracellular traps (NETs) that modulate aggregation and degradation of chemokines and cytokines and partitioning of crystallized urate against immune cells. Calcium pyrophosphate dehydrate (CPP) crystals are formed in the cartilage and CPP deposition can be found in 30% of people aged over 80 years. Inorganic pyrophosphate (PPi) is synthesized in chondrocytes and plays an important part in the formation of calcium pyrophosphate crystals. The degradation is catalyzed by inorganic pyrophosphatases. If there is dysregulation of this homeostasis more PPi is produced, which ultimately contributes to the formation of the CPP crystals.

  16. Dual roles of borax in kinetics of calcium sulfate dihydrate formation.

    PubMed

    Jiang, Wenge; Pan, Haihua; Tao, Jinhui; Xu, Xurong; Tang, Ruikang

    2007-04-24

    An additive is not exclusively retardant or promoter for a crystallization system. The kinetic studies of calcium sulfate dihydrate (CSD) crystal growth demonstrated that borax played dual roles in the reaction, which accelerated CSD formations at the low concentration levels but inhibited the crystal growth at the high ones. In situ atomic force microscopy studies revealed that borax modulated the CSD crystallization via two different pathways: promoted the secondary nucleation to increase the step density on the growing crystal faces but simultaneously retarded the spread of these growth steps by the Langmuir adsorption. These two contradictory factors were incorporated in the crystallization, and their balance was regulated by the borax concentration. Both the macroscopic and microscopic experimental data nicely displayed the crystallization model of birth and spread that was able to account for the behaviors of borax in CSD formations.

  17. Crystallization of calcium sulfate dihydrate in the presence of some metal ions

    NASA Astrophysics Data System (ADS)

    Hamdona, Samia K.; Al Hadad, Umaima A.

    2007-02-01

    Crystallization of calcium sulfate dihydrate (CaSO 4·2H 2O gypsum) in sodium chloride solutions in the presence of some metal ions, and over a range of relative super-saturation has been studied. The addition of metal ions, even at relatively low concentration (10 -6 mol l -1), markedly retard the rate of crystallization of gypsum. Retardation effect was enhanced with increase in the additives contents. Moreover, the effect was enhanced as the relative super-saturation decreases. Influence of mixed additives on the rate of crystallization (Cd 2++Arg, Cd 2++H 3PO 4 and Cd 2++PAA) has also been studied. Direct adsorption experiments of these metal ions on the surface of gypsum crystals have been made for comparison.

  18. Gitelman syndrome disclosed by calcium pyrophosphate deposition disease: early diagnosis by ultrasonographic study.

    PubMed

    Zabotti, A; Della Siega, P; Picco, L; Quartuccio, L; Bassetti, M; De Vita, S

    2016-06-23

    Gitelman's syndrome is a rare autosomal-recessive tubular disorder characterized by hypomagnesemia and hypocalciuria associated to hypokalemia. The clinical spectrum is wide and usually characterized by chronic fatigue, cramps, muscle weakness and paresthesiae. We describe a case of a 43 year-old male patient with early onset of knee arthritis and no other symptoms. Ultrasound revealed diffuse and confluent hyperechoic deposits in cartilage, fibrocartilage of the menisci and synovium and calcium pyrophosphate crystals were observed in the synovial fluid of the knee. The concomitant presence of hypomagnesemia, hypocalciuria and hypokalemia made clear the diagnosis of Gitelman's syndrome associated with chondrocalcinosis.

  19. Diff Quik staining method for detection and identification of monosodium urate and calcium pyrophosphate crystals in synovial fluids

    PubMed Central

    Selvi, E; Manganelli, S; Catenaccio, M; De Stefano, R; Frati, E; Cucini, S; Marcolongo, R

    2001-01-01

    OBJECTIVE—To evaluate whether the Diff Quik (DQ) staining method might prove useful in identifying monosodium urate (MSU) and calcium pyrophosphate dihydrate (CPPD) crystals on permanent mounted stained slides.
METHODS—27 synovial fluid (SF) samples obtained from the knees of 21 patients with acute CPPD disease and 6 with acute gout were studied. Wet analysis for crystal detection and identification was performed within one hour of joint aspiration. In addition, 16 inflammatory synovial effusions obtained from patients with knee arthritis induced by non-crystalline inflammatory diseases were studied. For each SF, a DQ stained slide was analysed by two of the authors trained in SF analysis. The observers were blinded to the type of crystals present in the SF. Each slide was analysed by compensated polarised as well as transmitted light microscopy. An SF was considered positive if intracellular and/or extracellular crystals were clearly identified. In addition, the observer was asked to identify the type of the crystals using compensated polarised light microscopy. Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) of the DQ staining method were determined.
RESULTS—51 true positive and 28 true negative cases were correctly classified (39 CPPD samples, 12 MSU samples, 28 samples of crystal unrelated arthropathies). Overall, four false positive and three false negative cases were reported. In all the false positive cases, extracellular CPPD crystals were erroneously identified, whereas CPPD crystals present in the SF were not identified in the three false negative cases. All MSU specimens were correctly diagnosed. The overall specificity, sensitivity, and accuracy using DQ stained slides for crystal confirmation were respectively 87.5%, 94.4%, and 91.9%. The PPV was 92.7% and the NPV 90.3%. In particular, the specificity, sensitivity, and accuracy for CPPD detection were 90.9%, 92.9%, and 91

  20. High early strength calcium phosphate bone cement: effects of dicalcium phosphate dihydrate and absorbable fibers.

    PubMed

    Burguera, Elena F; Xu, Hockin H K; Takagi, Shozo; Chow, Laurence C

    2005-12-15

    Calcium phosphate cement (CPC) sets in situ to form resorbable hydroxyapatite with chemical and crystallographic similarity to the apatite in human bones, hence it is highly promising for clinical applications. The objective of the present study was to develop a CPC that is fast setting and has high strength in the early stages of implantation. Two approaches were combined to impart high early strength to the cement: the use of dicalcium phosphate dihydrate with a high solubility (which formed the cement CPC(D)) instead of anhydrous dicalcium phosphate (which formed the conventional cement CPC(A)), and the incorporation of absorbable fibers. A 2 x 8 design was tested with two materials (CPC(A) and CPC(D)) and eight levels of cement reaction time: 15 min, 30 min, 1 h, 1.5 h, 2 h, 4 h, 8 h, and 24 h. An absorbable suture fiber was incorporated into cements at 25% volume fraction. The Gilmore needle method measured a hardening time of 15.8 min for CPC(D), five-fold faster than 81.5 min for CPC(A), at a powder:liquid ratio of 3:1. Scanning electron microscopy revealed the formation of nanosized rod-like hydroxyapatite crystals and platelet crystals in the cements. At 30 min, the flexural strength (mean +/- standard deviation; n = 5) was 0 MPa for CPC(A) (the paste did not set), (4.2 +/- 0.3) MPa for CPC(D), and (10.7 +/- 2.4) MPa for CPC(D)-fiber specimens, significantly different from each other (Tukey's at 0.95). The work of fracture (toughness) was increased by two orders of magnitude for the CPC(D)-fiber cement. The high early strength matched the reported strength for cancellous bone and sintered porous hydroxyapatite implants. The composite strength S(c) was correlated to the matrix strength S(m): S(c) = 2.16S(m). In summary, substantial early strength was imparted to a moldable, self-hardening and resorbable hydroxyapatite via two synergistic approaches: dicalcium phosphate dihydrate, and absorbable fibers. The new fast-setting and strong cement may help prevent

  1. Synthesis of β-Calcium Pyrophosphate by sol-gel method

    NASA Astrophysics Data System (ADS)

    Windarti, T.; Taslimah; Haris, A.; Astuti, Y.; Darmawan, A.

    2017-02-01

    Beta calcium pyrophosphate [β-CPP, β-Ca2P2O7] can be used as bone graft extender in posterolateral lumbar fusion. In this research, β-CPP was synthesized by sol-gel method using phosphorus pentaoxide [P2O5] and calcium nitrate tetrahydrate [Ca(NO3)2.4H2O] as phosphorus and calcium precursors. The reaction was carried out in ethanol medium with Ca/P ratio of 1.67. After 21 hours of reaction and 20 hours of drying at 80°C, white powder of amorphous calcium phosphate (ACP) was produced. Transformation of ACP to β-CPP was undertaken by firring at 400-800°C for 8 hours. Transformations of amorphous to microcrystalline, semicrystalline and crystalline structures occur at 400, 600 and 800°C, respectively. The β-CPP with the crystallite size of 61.71 nm, Ca/P ratio of 0.89 and Ca/O ratio of 0.21 was achieved by firing at 800°C. Morphology changes due to firing in which irregular shape of β-CPP at 400° changed to regular cuboid at 600 °C and above.

  2. Definition and Reliability Assessment of Elementary Ultrasonographic Findings in Calcium Pyrophosphate Deposition Disease: A Study by the OMERACT Calcium Pyrophosphate Deposition Disease Ultrasound Subtask Force.

    PubMed

    Filippou, Georgios; Scirè, Carlo A; Damjanov, Nemanja; Adinolfi, Antonella; Carrara, Greta; Picerno, Valentina; Toscano, Carmela; Bruyn, George A; D'Agostino, Maria Antonietta; Delle Sedie, Andrea; Filippucci, Emilio; Gutierrez, Marwin; Micu, Mihaela; Möller, Ingrid; Naredo, Esperanza; Pineda, Carlos; Porta, Francesco; Schmidt, Wolfgang A; Terslev, Lene; Vlad, Violeta; Zufferey, Pascal; Iagnocco, Annamaria

    2017-03-01

    To define the ultrasonographic characteristics of calcium pyrophosphate crystal (CPP) deposits in joints and periarticular tissues and to evaluate the intra- and interobserver reliability of expert ultrasonographers in the assessment of CPP deposition disease (CPPD) according to the new definitions. After a systematic literature review, a Delphi survey was circulated among a group of expert ultrasonographers, who were members of the CPPD Ultrasound (US) Outcome Measures in Rheumatology (OMERACT) subtask force, to obtain definitions of the US characteristics of CPPD at the level of fibrocartilage (FC), hyaline cartilage (HC), tendon, and synovial fluid (SF). Subsequently, the reliability of US in assessing CPPD at knee and wrist levels according to the agreed definitions was tested in static images and in patients with CPPD. Cohen's κ was used for statistical analysis. HC and FC of the knee yielded the highest interobserver κ values among all the structures examined, in both the Web-based (0.73 for HC and 0.58 for FC) and patient-based exercises (0.55 for the HC and 0.64 for the FC). Kappa values for the other structures were lower, ranging from 0.28 in tendons to 0.50 in SF in the static exercise and from 0.09 (proximal patellar tendon) to 0.27 (triangular FC of the wrist) in the patient-based exercise. The new OMERACT definitions for the US identification of CPPD proved to be reliable at the level of the HC and FC of the knee. Further studies are needed to better define the US characteristics of CPPD and optimize the scanning technique in other anatomical sites.

  3. White mineral trioxide aggregate mixed with calcium chloride dihydrate: chemical analysis and biological properties

    PubMed Central

    2017-01-01

    Objectives This study aimed to evaluate the chemical and biological properties of fast-set white mineral trioxide aggregate (FS WMTA), which was WMTA combined with calcium chloride dihydrate (CaCl2·2H2O), compared to that of WMTA. Materials and Methods Surface morphology, elemental, and phase analysis were examined using scanning electron microscope (SEM), energy dispersive X-ray microanalysis (EDX), and X-ray diffraction (XRD), respectively. The cytotoxicity and cell attachment properties were evaluated on human periodontal ligament fibroblasts (HPLFs) using methyl-thiazol-diphenyltetrazolium (MTT) assay and under SEM after 24 and 72 hours, respectively. Results Results showed that the addition of CaCl2·2H2O to WMTA affected the surface morphology and chemical composition. Although FS WMTA exhibited a non-cytotoxic profile, the cell viability values of this combination were lesser than WMTA, and the difference was significant in 7 out of 10 concentrations at the 2 time intervals (p < 0.05). HPLFs adhered over the surface of WMTA and at the interface, after 24 hours of incubation. After 72 hours, there were increased numbers of HPLFs with prominent cytoplasmic processes. Similar findings were observed with FS WMTA, but the cells were not as confluent as with WMTA. Conclusions The addition of CaCl2·2H2O to WMTA affected its chemical properties. The favorable biological profile of FS WMTA towards HPLFs may have a potential impact on its clinical application for repair of perforation defects. PMID:28808634

  4. Characterization and in vitro evaluation of biphasic calcium pyrophosphate-tricalciumphosphate radio frequency magnetron sputter coatings.

    PubMed

    Takahashi, K; van den Beucken, J J J P; Wolke, J G C; Hayakawa, T; Nishiyama, N; Jansen, J A

    2008-03-01

    The objective of this study was to characterize the physicochemical, dissolution, and osteogenic properties of radio frequency magnetron sputtered dicalcium pyrophosphate/tricalciumphosphate (Pyro/TCP) and hydroxylapatite (HA) coatings. Therefore Pyro/TCP and HA coatings were deposited on grit-blasted titanium discs. The results showed that the deposited coatings were amorphous and changed into a crystalline structure after IR heat-treatment of 550 degrees C for HA and 650 degrees C for Pyro/TCP. Heat-treated HA coatings appeared to be stable during immersion in simulated body fluid (SBF), that is no changes in the XRD pattern were observed. Also, no dissolution of the coating was observed by scanning electron microscopy (SEM). Energy dispersive spectroscopy (EDS) revealed that the Ca/P ratio of the HA coatings remained constant during SBF immersion. On the other hand, the heat-treated Pyro/TCP coatings showed a surface reaction of calcium pyrophosphate into a beta-tricalcium phosphate phase during SBF immersion. This was confirmed by EDS analysis. Rat bone marrow-derived osteoblast-like cells cultured on the heat-treated substrates showed that cell proliferation and differentiation occurred on both types of bioceramic coatings. No significant differences for proliferation and early differentiation were observed between cells cultured on heat-treated Pyro/TCP and HA at individual time points. However, osteocalcin expression, a late marker for osteoblast-like cell differentiation, was significantly increased after 12 days of culture on HA-coatings. These results were confirmed by SEM observations and suggest increased osteogenic properties for HA-coatings over Pyro/TCP-coatings. Additional research is necessary to obtain conclusive evidence on the in vivo osteogenic capacity of Pyro/TCP coatings.

  5. Injectable and rapid-setting calcium phosphate bone cement with dicalcium phosphate dihydrate.

    PubMed

    Burguera, Elena F; Xu, Hockin H K; Weir, Michael D

    2006-04-01

    Calcium phosphate cement (CPC) sets in situ with intimate adaptation to the contours of defect surfaces, and forms an implant having a structure and composition similar to hydroxyapatite, the putative mineral in teeth and bones. The objective of the present study was to develop an injectable CPC using dicalcium phosphate dihydrate (DCPD) with a high solubility for rapid setting. Two agents were incorporated to impart injectability and fast-hardening to the cement: a hardening accelerator (sodium phosphate) and a gelling agent (hydroxypropyl methylcellulose, HPMC). The cement with DCPD was designated as CPC(D), and the conventional cement was referred to as CPC(A). Using water without sodium phosphate, CPC(A) had a setting time of 82 +/- 6 min. In contrast, CPC(D) exhibited rapid setting with a time of 17 +/- 1 min. At 0.2 mol/L sodium phosphate, setting time for CPC(D) was 15 +/- 1 min, significantly faster than 40 +/- 2 min for CPC(A) (Tukey's at 0.95). Sodium phosphate decreased the paste injectability (measured as the paste mass extruded from the syringe divided by the original paste mass inside the syringe). However, the addition of HPMC dramatically increased the paste injectability. For CPC(D), the injectability was increased from 65% +/- 12% without HPMC to 98% +/- 1% with 1% HPMC. Injectability of CPC(A) was also doubled to 99% +/- 1%. The injectable and rapid-setting CPC(D) possessed flexural strength and elastic modulus values overlapping the reported values for sintered porous hydroxyapatite implants and cancellous bone. In summary, the rapid setting and relatively high strength and elastic modulus of CPC(D) should help the graft to quickly attain strength and geometrical integrity within a short period of time postoperatively. Furthermore, the injectability of CPC(D) may have potential for procedures involving defects with limited accessibility or narrow cavities, when there is a need for precise placement of the paste, and when using minimally invasive

  6. Further characterization of ATP-initiated calcification by matrix vesicles isolated from rachitic rat cartilage. Membrane perturbation by detergents and deposition of calcium pyrophosphate by rachitic matrix vesicles.

    PubMed

    Hsu, H H; Camacho, N P; Anderson, H C

    1999-01-12

    Although membrane associated enzymes such as ATPase, alkaline phosphatase, and NTP pyrophosphohydrolase in matrix vesicles (MVs) may underlie the mechanisms of ATP-promoted calcification, prior to the current investigation, the role of the MV membrane in calcification had not been addressed. In this study, various perturbations were introduced to the MV membrane in in vitro calcification systems to determine ideal conditions for ATP-initiated calcification by MVs isolated from rachitic rat epiphyseal cartilage. Membrane integrity appears to be required, since the rupture of the vesicular membrane by vigorously mixing with 10% butanol abolished calcification. In contrast, a mild treatment of MVs with low concentrations (e.g., 0.01%, which is much below the critical concentration for micelle formation) of either neutral Triton X-100 or anionic deoxycholate stimulated calcification by >2-fold, without inducing obvious changes in vesicular appearance. Fourier transform infrared spectroscopic studies were done to identify the mineral phase formed in these experiments. For the first time, rachitic MVs were shown to induce the formation of a calcium pyrophosphate dihydrate-like phase after their exposure to calcifying medium with 1 mM ATP. The integration of spectral areas indicated that calcification was enhanced by Triton X-100. The detergent effect was reversible and appeared to be not mediated through activation of ATPase, alkaline phosphatase, or ATP pyrophosphohydrolase. In contrast to neutral Triton X-100 and anionic deoxycholate, cationic cetyltrimethylammonium bromide inhibited both ATPase activity (I50=10 microM) and ATP-initiated calcification. These observations suggest that membrane perturbations can affect calcification and that the presence of NTP-pyrophosphohydrolase in MVs may play a role in the deposition of CaPPi in rachitic cartilage.

  7. Calcium pyrophosphate and monosodium urate crystals in synovial fluid as a cause of pseudoeosinophilia.

    PubMed

    Robier, Christoph; Neubauer, Manfred; Quehenberger, Franz; Stettin, Mariana; Rainer, Franz

    2011-08-01

    Synovial fluid (SF) leukocytes can be counted microscopically in a Neubauer chamber or by automated procedures using haematology analysers. Knowledge of laboratory artefacts is crucial for the correct interpretation of results obtained using automated methods. SF pseudoeosinophilia has recently been described as a new artefact in patients with crystal-related arthropathies. We investigated whether pseudoeosinophilia of SF is restricted to crystal-related disorders, or if it may also occur in other arthropathies. We compared the percentages of eosinophils in 120 crystal containing SF samples with 185 crystal-free specimens using the Wilcoxon test. Crystal positive samples, determined by polarised microscopy, contained at least two monosodium urate or calcium pyrophosphate crystals per 10 high power fields (630× magnification). True SF eosinophilia was ruled out by microscopic examination of stained slides. Crystal positive samples had significantly higher percentages of eosinophils than the controls (p<0.0001). No significant differences between the two crystal types were found (p=0.693). Thus, pseudoeosinophilia was significantly correlated with the presence of crystals, and none of the distinct crystal types was more likely to be associated with pseudoeosinophilia. In this study, SF pseudoeosinophilia was confirmed as a crystal-related laboratory artefact which has to be considered in the interpretation of automated SF leukocyte differential counts.

  8. Carpal tunnel syndrome induced by two types of calcium deposition.

    PubMed

    Ikawa, H; Hashizume, H; Inoue, H

    1997-12-01

    Two rare cases of carpal tunnel syndrome caused by calcification in the carpal tunnel are reported. One case involved a tumorous calcification consisting of basic calcium phosphate, and the other involved a diffuse calcification consisting of a mixture of calcium pyrophosphate dihydrate and basic calcium phosphate. These cases suggest that the shape of carpal tunnel calcifications is influenced by the nature of calcifying substance itself, i.e., whether it is heterogenous or homogenous.

  9. Osteogenic biphasic calcium sulphate dihydrate/iron-modified alpha-tricalcium phosphate bone cement for spinal applications: in vivo study.

    PubMed

    Vlad, M D; Sindilar, E V; Mariñoso, M L; Poeată, I; Torres, R; López, J; Barracó, M; Fernández, E

    2010-02-01

    In this study, the biocompatibility and the osteogenic features of a new iron-modified alpha-tricalcium phosphate (IM/alpha-TCP) and calcium sulphate dihydrate (CSD) biphasic cement (IM/alpha-TCP/CSD-BC) have been investigated in terms of the in vivo cement resorption, bone tissue formation and host tissue response on sheep animal model. Histological evaluation performed on undecalcified cement-bone specimens assessed the in vivo behaviour. It has been shown that the new IM/alpha-TCP/CSD-BC has the ability to produce firm bone binding in vivo (i.e. bioactivity). Qualitative histology proved cement biocompatibility, osteoconduction and favourable resorption, mainly through a macrophage-mediated mechanism. The results showed that the new cements have biocompatible and osteogenic features of interest as possible cancellous bone replacement biomaterial for minimally invasive spinal surgery applications.

  10. Systematic review and quality analysis of emerging diagnostic measures for calcium pyrophosphate crystal deposition disease

    PubMed Central

    Wu, Y; Chen, K; Terkeltaub, R

    2016-01-01

    Objectives Calcium pyrophosphate crystal deposition disease (CPPD) is common, yet prevalence and overall clinical impact remain unclear. Sensitivity and specificity of CPPD reference standards (conventional crystal analysis (CCA) and radiography (CR)) were meta-analysed by EULAR (published 2011). Since then, new diagnostic modalities are emerging. Hence, we updated 2009–2016 literature findings by systematic review and evidence grading, and assessed unmet needs. Methods We performed systematic search of full papers (PubMed, Scopus/EMBASE, Cochrane 2009–2016 databases). Search terms included CPPD, chondrocalcinosis, pseudogout, ultrasound, MRI, dual energy CT (DECT). Paper selection, data abstraction, EULAR evidence level, and Quality Assessment of Diagnostic Accuracy Studies (QUADAS)-2 bias and applicability grading were performed independently by 3 authors. Results We included 26 of 111 eligible papers, which showed emergence in CPPD diagnosis of ultrasound (U/S), and to lesser degree, DECT and Raman spectroscopy. U/S detected CPPD crystals in peripheral joints with sensitivity >80%, superior to CR. However, most study designs, though analytical, yielded low EULAR evidence level. DECT was marginally explored for CPPD, compared with 35 published DECT studies in gout. QUADAS-2 grading indicated strong applicability of U/S, DECT and Raman spectroscopy, but high study bias risk (in ∼30% of papers) due to non-controlled designs, and non-randomised subject selection. Conclusions Though CCA and CR remain reference standards for CPPD diagnosis, U/S, DECT and Raman spectroscopy are emerging U/S sensitivity appears to be superior to CR. We identified major unmet needs, including for randomised, blinded, controlled studies of CPPD diagnostic performance and rigorous analyses of 4 T MRI and other emerging modalities. PMID:27933211

  11. Calcium-deficient apatite synthesized by ammonia hydrolysis of dicalcium phosphate dihydrate: influence of temperature, time, and pressure.

    PubMed

    Obadia, Laetitia; Rouillon, Thierry; Bujoli, Bruno; Daculsi, Guy; Bouler, Jean Michel

    2007-01-01

    In this work, calcium-deficient apatites (CDA) were synthesized by ammonia hydrolysis reaction of dicalcium phosphate dihydrate (DCPD; CaHPO4 x 2 H2O) to obtain biphasic calcium phosphates (BCP) without any extraionic substitution. The influence of three parameters was studied: temperature of the reaction (70 and 100 degrees C), time of the reaction (4 and 18 h), and the pressure (open and closed system). Experiments were made according to a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. Moreover, the influence of temperature (until 200 degrees C) was investigated. The crystal size of CDA was determined according to the Scherrer's formula and from Rietveld refinements taking the CDA anisotropy into account. The last method seems to be a reliable method to determine crystallite sizes of CDA, since crystallite sizes of CDA along <00l> and directions were accessible. The results describe the hydroxyapatite % (HA%) in BCP by a first-order polynomial equation in the experimental area studied and the HA content was found to increase by raising time and temperature of the reaction. Moreover, the type of reaction system (open/closed vessel) appeared to have little influence on HA%.

  12. Fabrication of Carbonate Apatite Block through a Dissolution-Precipitation Reaction Using Calcium Hydrogen Phosphate Dihydrate Block as a Precursor.

    PubMed

    Tsuru, Kanji; Yoshimoto, Ayami; Kanazawa, Masayuki; Sugiura, Yuki; Nakashima, Yasuharu; Ishikawa, Kunio

    2017-03-31

    Carbonate apatite (CO₃Ap) block, which is a bone replacement used to repair defects, was fabricated through a dissolution-precipitation reaction using a calcium hydrogen phosphate dihydrate (DCPD) block as a precursor. When the DCPD block was immersed in NaHCO₃ or Na₂CO₃ solution at 80 °C, DCPD converted to CO₃Ap within 3 days. β-Tricalcium phosphate was formed as an intermediate phase, and it was completely converted to CO₃Ap within 2 weeks when the DCPD block was immersed in Na₂CO₃ solution. Although the crystal structures of the DCPD and CO₃Ap blocks were different, the macroscopic structure was maintained during the compositional transformation through the dissolution-precipitation reaction. CO₃Ap block fabricated in NaHCO₃ or Na₂CO₃ solution contained 12.9 and 15.8 wt % carbonate, respectively. The diametral tensile strength of the CO₃Ap block was 2 MPa, and the porosity was approximately 57% regardless of the carbonate solution. DCPD is a useful precursor for the fabrication of CO₃Ap block.

  13. Time-dependent subcellular structure injuries induced by nano-/micron-sized calcium oxalate monohydrate and dihydrate crystals.

    PubMed

    Sun, Xin-Yuan; Yu, Kai; Ouyang, Jian-Ming

    2017-10-01

    Comparative studies were conducted to investigate the time effect of cell injury induced by nano-sized (50nm) and micron-sized (10μm) calcium oxalate monohydrate (COM) and dihydrate (COD) crystals in African green monkey renal epithelial (Vero) cells. The effects of nano-/micron-sized COM and COD exposure on Vero cells were investigated by detecting the cell viability, cell morphology, LDH release, reactive oxygen species, mitochondrial membrane potential, cell cycle, and cell apoptosis, as well as the intracellular and extracellular crystal distribution. Nano-/micron-sized COM and COD exposure lead to subcellular organelle injury in varying degrees, but the injury sequence of various organelles differed. The time sequence of organelle injury presenting significant variation was described as follows: cell membrane injury (1h)

  14. Biphasic calcium sulfate dihydrate/iron-modified alpha-tricalcium phosphate bone cement for spinal applications: in vitro study.

    PubMed

    Vlad, M D; Valle, L J; Poeată, I; López, J; Torres, R; Barracó, M; Fernández, E

    2010-04-01

    In this study, the cytocompatibility of new 'iron-modified/alpha-tricalcium phosphate (IM/alpha-TCP) and calcium sulfate dihydrate (CSD)' bone cement (IM/alpha-TCP/CSD-BC) intended for spinal applications has been approached. The objective was to investigate by direct-contact osteoblast-like cell cultures (from 1 to 14 days) the in vitro cell adhesion, proliferation, morphology and cytoskeleton organization of MG-63 cells seeded onto the new cements. The results were as follows: (a) quantitative MTT-assay and scanning electron microscopy (SEM) showed that cell adhesion, proliferation and viability were not affected with time by the presence of iron in the cements; (b) double immunofluorescent labeling of F-actin and alpha-tubulin showed a dynamic interaction between the cell and its porous substrates sustaining the locomotion phenomenon on the cements' surface, which favored the colonization, and confirming the biocompatibility of the experimental cements; (c) SEM-cell morphology and cytoskeleton observations also evidenced that MG-63 cells were able to adhere, to spread and to attain normal morphology on the new IM/alpha-TCP/CSD-BC which offered favorable substratum properties for osteoblast-like cells proliferation and differentiation in vitro. The results showed that these new iron-modified cement-like biomaterials have cytocompatible features of interest not only as possible spinal cancellous bone replacement biomaterial but also as bone tissue engineering scaffolds.

  15. Effect of pyrophosphate ions on the conversion of calcium-lithium-borate glass to hydroxyapatite in aqueous phosphate solution.

    PubMed

    Fu, Hailuo; Rahaman, Mohamed N; Day, Delbert E; Huang, Wenhai

    2010-10-01

    The conversion of glass to a hydroxyapatite (HA) material in an aqueous phosphate solution is used as an indication of the bioactive potential of the glass, as well as a low temperature route for preparing biologically useful materials. In this work, the effect of varying concentrations of pyrophosphate ions in the phosphate solution on the conversion of a calcium-lithium-borate glass to HA was investigated. Particles of the glass (150-355 μm) were immersed for up to 28 days in 0.25 M K(2)HPO(4) solution containing 0-0.1 M K(4)P(2)O(7). The kinetics of degradation of the glass particles and their conversion to HA were monitored by measuring the weight loss of the particles and the ionic concentration of the solution. The structure and composition of the conversion products were analyzed using X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopy. For K(4)P(2)O(7) concentrations of up to 0.01 M, the glass particles converted to HA, but the time for complete conversion increased from 2 days (no K(4)P(2)O(7)) to 10 days (0.01 M K(4)P(2)O(7)). When the K(4)P(2)O(7) concentration was increased to 0.1 M, the product consisted of an amorphous calcium phosphate material, which eventually crystallized to a pyrophosphate product (predominantly K(2)CaP(2)O(7) and Ca(2)P(2)O(7)). The consequences of the results for the formation of HA materials and devices by the glass conversion route are discussed.

  16. Size-controlled preparation of α-calcium sulphate hemihydrate starting from calcium sulphate dihydrate in the presence of modifiers and the dissolution rate in simulated body fluid.

    PubMed

    Chen, Jianmin; Gao, Jun; Yin, Hengbo; Liu, Fanggang; Wang, Aili; Zhu, Yongqiang; Wu, Zhanao; Jiang, Tingshun; Qin, Daming; Chen, Bujun; Ji, Yuqin; Sun, Min

    2013-08-01

    Different-sized α-calcium sulphate hemihydrate (α-CSH) rods were hydrothermally prepared by converting calcium sulphate dihydrate at 110-140°C in the presence of MgCl2, sodium citrate (CANa), and sodium dodecyl benzene sulfonate (SDBS) as the modifiers. The α-CSH rods with the average diameters and the average lengths in the ranges of 2.6-5.2 and 17.5-33.1 μm, respectively, were tunably prepared. The presence of the modifiers favoured the formation of small-sized α-CSH rods. The effect of the modifiers on decreasing the diameters of α-CSH rods was in an order of SDBS>CANa>MgCl2. The dissolution rates of the different-sized α-CSH rods prepared at 140°C in simulated body fluid were in an order of α-CSH (CANa)>α-CSH (MgCl2)>α-CSH (reference)>α-CSH (SDBS). The naked and small-sized α-CSH rods had high dissolution rates. The adsorption of SDBS on the surfaces of α-CSH rods decreased their dissolution rates. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Additive concentration effects on dicalcium phosphate dihydrate cements prepared using monocalcium phosphate monohydrate and hydroxyapatite.

    PubMed

    Santa Cruz Chavez, Grace; Alge, Daniel L; Chu, Tien-Min Gabriel

    2011-12-01

    In our previous study, we investigated the setting time, mechanical properties and microstructure of dicalcium phosphate dihydrate cements prepared using monocalcium phosphate monohydrate (MCPM) and hydroxyapatite (HA). Despite the use of sodium citrate as a setting regulator, setting occurs rapidly in the MCPM/HA system and further studies on other retardants are needed. In the present study, sodium pyrophosphate and sulfuric acid were tested to evaluate their effectiveness in maintaining workability of the cement paste. MCPM/HA cements at a powder to liquid ratio of 1.0 with sodium pyrophosphate and sulfuric acid at 10, 25, 50, 75 and 100 mM were manufactured and studied based on their setting time, mechanical and porosity properties, phase composition, and microstructure. These measurements were compared to our previous data using sodium citrate. The results showed that the additives have a dose-dependent effect on the setting time. Their order of efficiency is sodium pyrophosphate > sodium citrate > sulfuric acid. However, the sulfuric acid group exhibited the highest compressive strength (CS) compared to the other groups. A lack of correlation between the CS and the porosity of the cements suggested that a mechanism other than porosity reduction was responsible for the CS increase. Since x-ray diffraction analysis did not indicate an effect on composition, explanations based on calcium sulfate dihydrate formation and changes in microstructure were proposed based on scanning electron micrograph observations.

  18. Reinjury risk of nano-calcium oxalate monohydrate and calcium oxalate dihydrate crystals on injured renal epithelial cells: aggravation of crystal adhesion and aggregation.

    PubMed

    Gan, Qiong-Zhi; Sun, Xin-Yuan; Bhadja, Poonam; Yao, Xiu-Qiong; Ouyang, Jian-Ming

    2016-01-01

    Renal epithelial cell injury facilitates crystal adhesion to cell surface and serves as a key step in renal stone formation. However, the effects of cell injury on the adhesion of nano-calcium oxalate crystals and the nano-crystal-induced reinjury risk of injured cells remain unclear. African green monkey renal epithelial (Vero) cells were injured with H2O2 to establish a cell injury model. Cell viability, superoxide dismutase (SOD) activity, malonaldehyde (MDA) content, propidium iodide staining, hematoxylin-eosin staining, reactive oxygen species production, and mitochondrial membrane potential (Δψm) were determined to examine cell injury during adhesion. Changes in the surface structure of H2O2-injured cells were assessed through atomic force microscopy. The altered expression of hyaluronan during adhesion was examined through laser scanning confocal microscopy. The adhesion of nano-calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) crystals to Vero cells was observed through scanning electron microscopy. Nano-COM and COD binding was quantitatively determined through inductively coupled plasma emission spectrometry. The expression of hyaluronan on the cell surface was increased during wound healing because of Vero cell injury. The structure and function of the cell membrane were also altered by cell injury; thus, nano-crystal adhesion occurred. The ability of nano-COM to adhere to the injured Vero cells was higher than that of nano-COD crystals. The cell viability, SOD activity, and Δψm decreased when nano-crystals attached to the cell surface. By contrast, the MDA content, reactive oxygen species production, and cell death rate increased. Cell injury contributes to crystal adhesion to Vero cell surface. The attached nano-COM and COD crystals can aggravate Vero cell injury. As a consequence, crystal adhesion and aggregation are enhanced. These findings provide further insights into kidney stone formation.

  19. Reinjury risk of nano-calcium oxalate monohydrate and calcium oxalate dihydrate crystals on injured renal epithelial cells: aggravation of crystal adhesion and aggregation

    PubMed Central

    Gan, Qiong-Zhi; Sun, Xin-Yuan; Bhadja, Poonam; Yao, Xiu-Qiong; Ouyang, Jian-Ming

    2016-01-01

    Background Renal epithelial cell injury facilitates crystal adhesion to cell surface and serves as a key step in renal stone formation. However, the effects of cell injury on the adhesion of nano-calcium oxalate crystals and the nano-crystal-induced reinjury risk of injured cells remain unclear. Methods African green monkey renal epithelial (Vero) cells were injured with H2O2 to establish a cell injury model. Cell viability, superoxide dismutase (SOD) activity, malonaldehyde (MDA) content, propidium iodide staining, hematoxylin–eosin staining, reactive oxygen species production, and mitochondrial membrane potential (Δψm) were determined to examine cell injury during adhesion. Changes in the surface structure of H2O2-injured cells were assessed through atomic force microscopy. The altered expression of hyaluronan during adhesion was examined through laser scanning confocal microscopy. The adhesion of nano-calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD) crystals to Vero cells was observed through scanning electron microscopy. Nano-COM and COD binding was quantitatively determined through inductively coupled plasma emission spectrometry. Results The expression of hyaluronan on the cell surface was increased during wound healing because of Vero cell injury. The structure and function of the cell membrane were also altered by cell injury; thus, nano-crystal adhesion occurred. The ability of nano-COM to adhere to the injured Vero cells was higher than that of nano-COD crystals. The cell viability, SOD activity, and Δψm decreased when nano-crystals attached to the cell surface. By contrast, the MDA content, reactive oxygen species production, and cell death rate increased. Conclusion Cell injury contributes to crystal adhesion to Vero cell surface. The attached nano-COM and COD crystals can aggravate Vero cell injury. As a consequence, crystal adhesion and aggregation are enhanced. These findings provide further insights into kidney stone

  20. Study of a hydraulic dicalcium phosphate dihydrate/calcium oxide-based cement for dental applications.

    PubMed

    el-Briak, Hasna; Durand, Denis; Nurit, Josiane; Munier, Sylvie; Pauvert, Bernard; Boudeville, Phillipe

    2002-01-01

    By mixing CaHPO(4) x 2H(2)O (DCPD) and CaO with water or sodium phosphate buffers as liquid phase, a calcium phosphate cement was obtained. Its physical and mechanical properties, such as compressive strength, initial and final setting times, cohesion time, dough time, swelling time, dimensional and thermal behavior, and injectability were investigated by varying different parameters such as liquid to powder (L/P) ratio (0.35-0.7 ml g(-1)), molar calcium to phosphate (Ca/P) ratio (1.67-2.5) and the pH (4, 7, and 9) and the concentration (0-1 M) of the sodium phosphate buffer. The best results were obtained with the pH 7 sodium phosphate buffer at the concentration of 0.75 M. With this liquid phase, physical and mechanical properties depended on the Ca/P and L/P ratios, varying from 3 to 11 MPa (compressive strength), 6 to 10 min (initial setting time), 11 to 15 min (final setting time), 15 to 30 min (swelling time), 7 to 20 min (time of 100% injectability). The dough or working time was over 16 min. This cement expanded during its setting (1.2-5 % according to Ca/P and L/P ratios); this would allow a tight filling. Given the mechanical and rheological properties of this new DCPD/CaO-based cement, its use as root canal sealing material can be considered as classical calcium hydroxide or ZnO/eugenol-based pastes, without or with a gutta-percha point.

  1. Dry mechanochemical synthesis of hydroxyapatites from dicalcium phosphate dihydrate and calcium oxide: a kinetic study.

    PubMed

    El Briak-BenAbdeslam, Hassane; Mochales, Carolina; Ginebra, Maria Pau; Nurit, Josiane; Planell, Josep A; Boudeville, Philippe

    2003-12-01

    Calcium phosphate ceramics have been used successfully as synthetic bone substitutes in orthopedics, dentistry, and maxillofacial surgery. One way of preparing these ceramics is the sintering of a calcium-deficient hydroxyapatite (CDHA), which can be obtained in different ways. Mechanochemistry is one possible means of synthesizing CDHA, with an expected molar calcium-to-phosphate (Ca/P) ratio +/- 0.005. The grinding can be carried out under dry or wet conditions. To optimize the experimental conditions of CDHA preparation by dry mechanosynthesis and for a better understanding of the DCPD/CaO mechanochemical reaction, we performed a kinetic study in which some of the experimental parameters were varied. Carried out with two different vertical rotating ball mills, this kinetic study showed that (1) experiments are reproducible and give as a final product a hydroxyapatite powder, formed of nano-sized crystals of around 20 nm, with a controlled Ca/P ratio; (2) the time for complete disappearance of DCPD and the time for complete reaction are in direct proportion to the mass of the ground powder; but (3) the time for complete disappearance of DCPD is independent of the Ca/P ratio while the time for complete reaction increases exponentially with the Ca/P ratio; and (4) the time for complete disappearance of DCPD corresponds to the time for complete reaction solely for Ca/P = 1.5. These observations suggest a reaction mechanism in two well differentiated stages: (First stage) CaO reacts with DCPD to give first an amorphous calcium phosphate (ACP) with a low Ca/P ratio that transforms into CDHA when its Ca/P ratio reaches 1.5. At the same time, CaO is hydrated into Ca(OH)(2) by the water produced by the reaction. (Second stage) If the Ca/P > 1.5 in the initial mixture, the excess Ca(OH)(2) is added to CDHA 1.5 by reacting with the HPO(4) group of CDHA until its Ca/P ratio reaches the expected value. The slower the reaction, the higher the Ca/P in the initial mixture.

  2. Phase transformation of calcium oxalate dihydrate-monohydrate: Effects of relative humidity and new spectroscopic data

    NASA Astrophysics Data System (ADS)

    Conti, Claudia; Casati, Marco; Colombo, Chiara; Realini, Marco; Brambilla, Luigi; Zerbi, Giuseppe

    2014-07-01

    New data on vibrational properties of calcium oxalates and their controversial transformation mechanism are presented. We have focused on whewellite (CaC2O4·H2O) and weddellite [CaC2O4·(2 + x) H2O], the most common phases of calcium oxalate; these compounds occur in many organisms, in kidney stones and in particular kinds of films found on the surface of many works of art. Low temperature experiments carried out by Fourier transform infrared spectroscopy have highlighted both the high structural order in the crystalline state of whewellite and the disordered distribution of the zeolitic water molecules in weddellite. The synthesised nanocrystals of weddellite have been kept under different hygrometric conditions in order to study, by X-ray powder diffraction, the role of “external” water molecules on their stability. Moreover, in order to identify the different kinds of water molecules, a re-investigation, supported by quantum chemical calculations, of the observed vibrational spectra (IR and Raman) of whewellite has been conducted.

  3. Thermal analysis of calcium sulfate dihydrate sources used to manufacture gypsum wallboard

    SciTech Connect

    Engbrecht, Dick C.; Hirschfeld, Deidre A.

    2016-07-27

    Gypsum wallboard has been used for over 100 years as a barrier to the spread of fire in residential and commercial structures. The gypsum molecule, CaSO4·2H2O, provides two crystalline waters that are released upon heating providing an endothermic effect. Manufacturers have recognized that the source of the gypsum ore is a factor that affects all aspects of its performance; thus, it is hypothesized that the impurities present in the gypsum ore are the causes of the performance differences. Differential Thermal Analysis/Thermogravimetric Analysis (DTA/TGA) and X-ray Diffraction (XRD) were used in this paper to compare and characterize samples of gypsum ore representing sources of natural, synthetic from a Flue Gas Desulfurization process (FGD) and blends thereof. The hemihydrate phase of representative natural, FGD, and reagent grade calcium sulfate were rehydrated with distilled water and evaluated by DTA/TGA. Analysis of the data shows distinct areas of similarity separated by the conversion to anhydrite ~250 °C. Compositional reconstructions based on DTA/TGA and XRD data were compared and although, the results were comparable, the DTA/TGA suggests thermally active compounds that were not detected by XRD. Anhydrite, silica and halite were reported by XRD but were not thermally reactive in the temperature range evaluated by DTA/TGA (ambient to 1050 °C). Finally, the presence of carbonate compounds (e.g., calcite and dolomite) were indicated by XRD and estimated from the thermal decomposition reaction ~700 °C.

  4. Thermal analysis of calcium sulfate dihydrate sources used to manufacture gypsum wallboard

    DOE PAGES

    Engbrecht, Dick C.; Hirschfeld, Deidre A.

    2016-07-27

    Gypsum wallboard has been used for over 100 years as a barrier to the spread of fire in residential and commercial structures. The gypsum molecule, CaSO4·2H2O, provides two crystalline waters that are released upon heating providing an endothermic effect. Manufacturers have recognized that the source of the gypsum ore is a factor that affects all aspects of its performance; thus, it is hypothesized that the impurities present in the gypsum ore are the causes of the performance differences. Differential Thermal Analysis/Thermogravimetric Analysis (DTA/TGA) and X-ray Diffraction (XRD) were used in this paper to compare and characterize samples of gypsum oremore » representing sources of natural, synthetic from a Flue Gas Desulfurization process (FGD) and blends thereof. The hemihydrate phase of representative natural, FGD, and reagent grade calcium sulfate were rehydrated with distilled water and evaluated by DTA/TGA. Analysis of the data shows distinct areas of similarity separated by the conversion to anhydrite ~250 °C. Compositional reconstructions based on DTA/TGA and XRD data were compared and although, the results were comparable, the DTA/TGA suggests thermally active compounds that were not detected by XRD. Anhydrite, silica and halite were reported by XRD but were not thermally reactive in the temperature range evaluated by DTA/TGA (ambient to 1050 °C). Finally, the presence of carbonate compounds (e.g., calcite and dolomite) were indicated by XRD and estimated from the thermal decomposition reaction ~700 °C.« less

  5. A comparison of the binding of urinary calcium oxalate monohydrate and dihydrate crystals to human kidney cells in urine

    PubMed Central

    Wang, Tingting; Thurgood, Lauren A.; Grover, Phulwinder K.; Ryall, Rosemary L.

    2010-01-01

    Objective To compare the binding kinetics of urinary calcium oxalate monohydrate (COM) and dihydrate (COD) crystals to human kidney (HK-2) cells in ultra-filtered (UF), and centrifuged and filtered (CF) human urine; and to quantify the binding of COM and COD crystals to cultured HK-2 cells in UF and CF urine samples collected from different individuals. Materials and methods Urine was collected from healthy subjects, pooled, centrifuged and filtered. 14C-oxalate-labelled COM and COD crystals were precipitated from the urine by adding oxalate after adjustment of two aliquots of the urine to 2 and 8 mm Ca2+, respectively. For the kinetic study, the crystals were incubated with HK-2 cells for up to 120 min in pooled CF urine adjusted to 2 and 8 mm Ca2+. Identical experiments were also carried out in UF urine samples collected from the same individuals. For the quantitative study, the same radioactively labelled COM and COD crystals were incubated with HK-2 cells for 50 min in separate CF and UF urines collected from eight healthy individuals at the native Ca2+ concentrations of the urines. Field emission electron microscopy and Fourier transform-infrared spectroscopy were used to confirm crystal morphology. Results Binding of both COM and COD crystals generally bound more strongly at 8 mm than at 2 mm Ca2+. The kinetic binding curves of COM were smooth, while those of COD were consistently biphasic, suggesting that the two crystal types induce different cellular metabolic responses: HK-2 cells crystals appear to possess a transitory mechanism for detaching COD, but not COM crystals. In UF urine, COM binding was significantly greater than that of COD at 2 mm Ca2+, but at 8 mm Ca2+ the binding of COD was greater at early and late time points. COD also bound significantly more strongly at early time points in CF urine at both 2 and 8 mm Ca2+. In both CF and UF urine, there was no difference between the binding affinity of urinary COM and COD crystals. Conclusion

  6. A comparison of the binding of urinary calcium oxalate monohydrate and dihydrate crystals to human kidney cells in urine.

    PubMed

    Wang, Tingting; Thurgood, Lauren A; Grover, Phulwinder K; Ryall, Rosemary L

    2010-12-01

    To compare the binding kinetics of urinary calcium oxalate monohydrate (COM) and dihydrate (COD) crystals to human kidney (HK-2) cells in ultra-filtered (UF), and centrifuged and filtered (CF) human urine; and to quantify the binding of COM and COD crystals to cultured HK-2 cells in UF and CF urine samples collected from different individuals. Urine was collected from healthy subjects, pooled, centrifuged and filtered. (14) C-oxalate-labelled COM and COD crystals were precipitated from the urine by adding oxalate after adjustment of two aliquots of the urine to 2 and 8 mm Ca(2+), respectively. For the kinetic study, the crystals were incubated with HK-2 cells for up to 120 min in pooled CF urine adjusted to 2 and 8 mm Ca(2+). Identical experiments were also carried out in UF urine samples collected from the same individuals. For the quantitative study, the same radioactively labelled COM and COD crystals were incubated with HK-2 cells for 50 min in separate CF and UF urines collected from eight healthy individuals at the native Ca(2+) concentrations of the urines. Field emission electron microscopy and Fourier transform-infrared spectroscopy were used to confirm crystal morphology. COM and COD crystals generally bound more strongly at 8 mm than at 2 mm Ca(2+). The kinetic binding curves of COM were smooth, while those of COD were consistently biphasic, suggesting that the two crystal types induce different cellular metabolic responses: HK-2 cells crystals appear to possess a transitory mechanism for detaching COD, but not COM crystals. In UF urine, COM binding was significantly greater than that of COD at 2 mm Ca(2+), but at 8 mm Ca(2+) the binding of COD was greater at early and late time points. COD also bound significantly more strongly at early time points in CF urine at both 2 and 8 mm Ca(2+). In both CF and UF urine, there was no difference between the binding affinity of urinary COM and COD crystals. Binding of both COM and COD crystals to cultured human

  7. The prevalence of monosodium urate and calcium pyrophosphate crystals in synovial fluid from wrist and finger joints.

    PubMed

    Galozzi, Paola; Oliviero, Francesca; Frallonardo, Paola; Favero, Marta; Hoxha, Ariela; Scanu, Anna; Lorenzin, Mariagrazia; Ortolan, Augusta; Punzi, Leonardo; Ramonda, Roberta

    2016-03-01

    The aim of this study was to assess the frequency of monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals in synovial fluids (SFs) aspirated from wrist and finger joints of patients with previously diagnosed joint diseases. We reviewed the results of SF analysis of 1593 samples and identified 126 patients with effusions in the small joints of the hands and wrists. We reported from patients' medical files data about sex, age, diagnosis, disease duration and the microscopic SF results. The prevalence of CPP crystals in SF was 85.71% in CPP-crystals arthritis (CPP-CA), 19.35% in rheumatoid arthritis (RA), 13.89% in osteoarthritis (OA) and 0% in psoriatic arthritis (PsA), spondyloarthritis (SpA), gout and miscellanea. The prevalence of MSU crystals in SF was 83.3% in gout, 10% in PsA, 2.8% in OA and 0% in RA, SpA, miscellanea and CPP-CA. Consistent with previously reported data concerning the big joints, microcrystals can be frequently found also in the small joints of patients with previous diagnosis. The finding underlines the importance of analyzing SF from the hand and wrist joints in the attempt to identify comorbidities associated with the presence of crystals and to develop targeted treatment strategies.

  8. Enzymes Involved in Pyrophosphate and Calcium Metabolism as Targets for Anti-scuticociliate Chemotherapy.

    PubMed

    Mallo, Natalia; Lamas, Jesús; DeFelipe, Ana-Paula; Sueiro, Rosa-Ana; Fontenla, Francisco; Leiro, José-Manuel

    2016-07-01

    Inorganic pyrophosphate (PPi) is a key metabolite in cellular bioenergetics under chronic stress conditions in prokaryotes, protists and plants. Inorganic pyrophosphatases (PPases) are essential enzymes controlling the cellular concentration of PPi and mediating intracellular pH and Ca(2+) homeostasis. We report the effects of the antimalarial drugs chloroquine (CQ) and artemisinin (ART) on the in vitro growth of Philasterides dicentrarchi, a scuticociliate parasite of turbot; we also evaluated the action of these drugs on soluble (sPPases) and vacuolar H+-PPases (H+-PPases). CQ and ART inhibited the in vitro growth of ciliates with IC50 values of respectively 74 ± 9 μM and 80 ± 8 μM. CQ inhibits the H+ translocation (with an IC50 of 13.4 ± 0.2 μM), while ART increased translocation of H+ and acidification. However, both drugs caused a decrease in gene expression of H+-PPases. CQ significantly inhibited the enzymatic activity of sPPases, decreasing the consumption of intracellular PPi. ART inhibited intracellular accumulation of Ca(2+) induced by ATP, indicating an effect on the Ca(2+) -ATPase. The results suggest that CQ and ART deregulate enzymes associated with PPi and Ca(2+) metabolism, altering the intracellular pH homeostasis vital for parasite survival and providing a target for the development of new drugs against scuticociliatosis. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

  9. Synovial Fluid Findings and Demographic Analysis of Patients With Coexistent Intra-articular Monosodium Urate and Calcium Pyrophosphate Crystals.

    PubMed

    Heselden, Emilia L; Freemont, Antony J

    2016-03-01

    The aim of this study was to investigate the characteristics of arthritis in which monosodium urate (urate) and calcium pyrophosphate (CPP) crystals coexisted in synovial fluid (SF) to aid patient management and set a baseline from which to investigate the pathophysiological basis of an unusual coexistence of 2 disorders. Synovial fluid analyses of 33,000 patients were reviewed, identifying those containing urate and/or CPP crystals. Synovial fluid cell count and differential cell count, together with patient age and gender, were retrieved from a computerized database spanning 22 years of SF analysis. In 6983 consecutive SF samples containing crystals, CPP crystals were found in 3685 (53%), urate in 3127 (44.5%), and both in 171 (2.5%). These 171 cases were deemed to have a mixed crystal arthropathy (MCA). Patients with MCA were 77% male and 23% female, and the highest incidence was found in those aged 76 to 80 years.Most commonly (69.4% of cases of MCA), high numbers (>20/10 high-power field) of both crystals and an acute inflammatory cell count were found. In the remainder, other patterns of crystals and cells were observed, perhaps suggesting different clinical situations in which these crystals coexist. This study presents evidence showing that with careful microscopic analysis the coexistence of urate and CPP crystals in a single joint is found in 2.5% of cases of crystal arthritis. The different patterns of SF findings and patient demography described here are novel and might have implications for patient management.

  10. Evaluation of host inflammatory responses of β-tricalcium phosphate bioceramics caused by calcium pyrophosphate impurity using a subcutaneous model.

    PubMed

    Lin, Kaili; Yuan, Wei; Wang, Lu; Lu, Jianxi; Chen, Lei; Wang, Zhen; Chang, Jiang

    2011-11-01

    Implantation of synthetic materials into body elicits inflammatory host responses that limit medical device integration and biological performance. Since the effective use of biomaterials in vivo requires good biocompatibility and bio-functionality, it is vital that we assess the inflammatory reactions provoked by various implanted biomaterials. In chemical precipitation of β-tricalcium phosphate [β-Ca₃(PO₄)₂, β-TCP], the impurity of calcium pyrophosphate (Ca₂P₂O₇, CPP) will easily appear if the preparation conditions are not well controlled. To test the influences of CCP-impurity on the biocompatibility of the material, four groups of β-TCP ceramic samples doped with 0.5-10 wt % of CCP impurity, and pure β-TCP and CCP samples were fabricated and implanted in rat subcutaneous site for one, two, and four weeks. The host tissue responses to the ceramics were evaluated by histomorphometric analysis, and the results were compared with pure β-TCPbioceramics. The results show that the CPP impurity can elicit and stimulate the inflammatory responses at the tissue/implant interface. Moreover, with the increase of CPP doping amount, the inflammation increases apparently. However, the pure β-TCP bioceramics only present slight post-implantation inflammatory responses. The influence of the CPP doping on the inflammatory responses is mainly related to a microparticles release because of an insufficient sintering of β-TCP by CPP doping. The microparticle release could be at the origin of local inflammation and cell/tissue damages. Therefore, to obtain perfect biocompatibility and high quality β-TCP bioceramics, it is important to avoid and control the CPP impurity in the preparation of β-TCP powders and bioceramics. 2011 Wiley Periodicals, Inc.

  11. Urinary stone formation: Efficacy of seed extract of Ensete superbum (Roxb.) Cheesman on growth inhibition of calcium hydrogen phosphate dihydrate crystals

    NASA Astrophysics Data System (ADS)

    Diana, K. J.; George, K. V.

    2013-01-01

    The effect of aqueous seed extract of Ensete superbum (Roxb.) Cheesman on in vitro crystallization and growth patterns of calcium hydrogen phosphate dihydrate (CaHPO4·2H2O, CHPD) crystals was studied using single diffusion gel growth technique. Reduction in growth of CHPD crystals was noticed with increasing concentrations of seed extract. The morphology of CHPD or brushite crystals was studied by microscopy. The structural changes of the treated crystals were assessed by SEM, FT-IR, XRD and TGA/DTA analysis. It is expected that this multidisciplinary approach for in vitro crystallization and characterization of CHPD crystals will provide a better explanation to develop novel strategies for prevention of urinary stones.

  12. Crystal arthritides - gout and calcium pyrophosphate arthritis : Part 2: clinical features, diagnosis and differential diagnostics.

    PubMed

    Schlee, S; Bollheimer, L C; Bertsch, T; Sieber, C C; Härle, P

    2017-02-23

    Gout develops in four stages beginning with an asymptomatic increase in blood levels of uric acid. An acute gout attack is an expression of an underlying inflammatory process, which in the course of time is self-limiting. Without therapy monosodium urate crystals remain in the synovial fluid and synovial membrane and trigger more acute attacks. In the course of the disease monosodium urate crystals form deposits (tophi) leading in severe forms to irreversible joint deformities with loss of functionality. In 20% of cases gout leads to involvement of the kidneys. Overproduction of uric acid can cause nephrolithiasis. These stones can be composed of uric acid or calcium phosphate. Another form of kidney disease caused by gout is uric acid nephropathy. This is a form of abacterial chronic inflammatory response with deposition of sodium urate crystals in the medullary interstitium. Acute obstructive nephropathy is relatively rare and characterized by renal failure due to uric acid precipitation in the tubules because of rapid cell lysis that occurs, for example, with chemotherapy. There is a causal interdependence between the occurrence of hyperuricemia and hypertension. Uric acid activates the renin-angiotensin-aldosterone (RAA) system and inhibits nitric oxide (NO) with the possible consequence of a rise in systemic vascular resistance or arteriolar vasculopathy; however, uric acid is also an apparently independent risk factor for atherosclerosis. In contrast to young patients, the diagnosis of an acute gout attack in the elderly can be a challenge for the physician. Polyarticular manifestations and obscure symptoms can make it difficult to differentiate it from rheumatoid arthritis and calcium pyrophosphate deposition disease (CPPD). Aspiration of synovial fluid with visualization of urate crystals using compensated polarized light microscopy is the gold standard for diagnosis of acute gout. Moreover, analysis of synovial fluid enables a distinction from septic

  13. Properties of fujicalin, a new modified anhydrous dibasic calcium phosphate for direct compression: comparison with dicalcium phosphate dihydrate.

    PubMed

    Schlack, H; Bauer-Brandl, A; Schubert, R; Becker, D

    2001-09-01

    The novel, commercially available, free-flowing spherically granulated dicalcium phosphate anhydrous (SGDCPA) Fujicalin for direct tableting was compared with directly compressible dicalcium phosphate dihYdrate (DCPD), the properties of which are well known. The two excipients were investigated and compared with regard to their physical and powder properties, compressibility, and compactibility. As a consequence of the spherical shape of its particles, SGDCPA shows the same good flowability and even better compactibility. In contrast to DCPD, SGDCPA shows significant uptake of moisture when exposed to relative humidities (RHs) exceeding 70%. For both excipients, the main deformation mechanism is fragmentation, with SGDCPA yielding significantly stronger tablets.

  14. Calcium Pyrophosphate Deposition (CPPD)

    MedlinePlus

    ... Walk Test (SMWT) Arthritis Impact Measurement Scales (AIMS) Evidence Based Practice (EBP) Fibromyalgia Impact Questionnaire (FIQ) Fracture Risk ... Investigators Resources for Doctoral Students/Post-Doctoral Fellows Evidence-Based Practice for Academic Researchers Responsible Data Management in ...

  15. Calcium pyrophosphate arthritis

    MedlinePlus

    ... disease that can cause attacks of arthritis. Like gout, crystals form in the joints. But in this ... CPPD arthritis can be confused with: Gouty arthritis (gout) Osteoarthritis Rheumatoid arthritis Exams and Tests Most arthritic ...

  16. Eight years of follow-up after laminectomy of calcium pyrophosphate crystal deposition in the cervical yellow ligament of patient with Coffin–Lowry syndrome

    PubMed Central

    Morino, Tadao; Ogata, Tadanori; Horiuchi, Hideki; Yamaoka, Shintaro; Fukuda, Mitsumasa; Miura, Hiromasa

    2016-01-01

    Abstract Background: We report 8 years of follow-up after decompression to treat cervical myelopathy in a patient with Coffin–Lowry syndrome (CLS). CLS is a rare X-linked semidominant syndrome associated with growth and psychomotor retardation, general hypotonia, and skeletal abnormalities. In this patient, the spinal cord was compressed by calcium pyrophosphate crystal deposition in the cervical yellow ligament (YL). To date, only 1 report has described clinical features after surgery for calcified cervical YL in CLS. Methods: A 15-year-old male with tetraplegia secondary to compression of the cervical spinal cord induced by a hypoplastic posterior arch of C1 and calcification of the YL from C2 to C7 was treated surgically with laminectomy from C1 to C7. The patient's history, clinical examination, imaging findings, and treatment are reported. The patient was incapable of speech because of mental retardation, so he could not describe his symptoms. Gait disturbance worsened over the 2 months before admission to our hospital. At admission, the patient could not move his extremities, and tendon reflexes of the upper and lower extremities were significantly increased. Computed tomography of the cervical spine showed YL calcification from C2 to C7. Magnetic resonance imaging showed consecutive compression of the cervical spinal cord. We diagnosed quadriplegia secondary to cervical cord damage and performed emergency surgery. Results: During C1–C7 laminectomy, YL calcification in C2–C7 was observed. The calcification was confirmed as calcium pyrophosphate by crystal analysis. Quadriplegia gradually resolved, and almost disappeared by 2 weeks after the operation. Cervical hyperlordosis was observed in radiographs starting from 1 month after the operation, but it has not progressed and is not associated with any symptoms. Conclusions: The efficacy of decompression continued, and no postoperative complications have occurred during at least 8 years of follow-up. PMID

  17. The effect of intracrystalline and surface-bound osteopontin on the degradation and dissolution of calcium oxalate dihydrate crystals in MDCKII cells.

    PubMed

    Thurgood, Lauren A; Sørensen, Esben S; Ryall, Rosemary L

    2012-02-01

    In vivo, urinary crystals are associated with proteins located within the mineral bulk as well as upon their surfaces. Proteins incarcerated within the mineral phase of retained crystals could act as a defence against urolithiasis by rendering them more vulnerable to destruction by intracellular and interstitial proteases. The aim of this study was to examine the effects of intracrystalline and surface-bound osteopontin (OPN) on the degradation and dissolution of urinary calcium oxalate dihydrate (COD) crystals in cultured Madin Darby canine kidney (MDCK) cells. [(14)C]-oxalate-labelled COD crystals with intracrystalline (IC), surface-bound (SB) and IC + SB OPN, were generated from ultrafiltered (UF) urine containing 0, 1 and 5 mg/L human milk OPN and incubated with MDCKII cells, using UF urine as the binding medium. Crystal size and degradation were assessed using field emission scanning electron microscopy (FESEM) and dissolution was quantified by the release of radioactivity into the culture medium. Crystal size decreased directly with OPN concentration. FESEM examination indicated that crystals covered with SB OPN were more resistant to cellular degradation than those containing IC OPN, whose degree of disruption appeared to be related to OPN concentration. Whether bound to the crystal surface or incarcerated within the mineral interior, OPN inhibited crystal dissolution in direct proportion to its concentration. Under physiological conditions OPN may routinely protect against stone formation by inhibiting the growth of COD crystals, which would encourage their excretion in urine and thereby perhaps partly explain why, compared with calcium oxalate monohydrate crystals, COD crystals are more prevalent in urine, but less common in kidney stones.

  18. Treatment with pyrophosphate inhibits uremic vascular calcification.

    PubMed

    O'Neill, W Charles; Lomashvili, Koba A; Malluche, Hartmut H; Faugere, Marie-Claude; Riser, Bruce L

    2011-03-01

    Pyrophosphate, which may be deficient in advanced renal failure, is a potent inhibitor of vascular calcification. To explore its use as a potential therapeutic, we injected exogenous pyrophosphate subcutaneously or intraperitoneally in normal rats and found that their plasma pyrophosphate concentrations peaked within 15 min. There was a single exponential decay with a half-life of 33 min. The kinetics were indistinguishable between the two routes of administration or in anephric rats. The effect of daily intraperitoneal pyrophosphate injections on uremic vascular calcification was then tested in rats fed a high-phosphate diet containing adenine for 28 days to induce uremia. Although the incidence of aortic calcification varied and was not altered by pyrophosphate, the calcium content of calcified aortas was significantly reduced by 70%. Studies were repeated in uremic rats given calcitriol to produce more consistent aortic calcification and treated with sodium pyrophosphate delivered intraperitoneally in a larger volume of glucose-containing solution to prolong plasma pyrophosphate levels. This maneuver significantly reduced both the incidence and amount of calcification. Quantitative histomorphometry of bone samples after double-labeling with calcein indicated that there was no effect of pyrophosphate on the rates of bone formation or mineralization. Thus, exogenous pyrophosphate can inhibit uremic vascular calcification without producing adverse effects on bone.

  19. Treatment with pyrophosphate inhibits uremic vascular calcification

    PubMed Central

    O’Neill, W. Charles; Lomashvili, Koba A.; Malluche, Hartmut H.; Faugere, Marie-Claude; Riser, Bruce L.

    2011-01-01

    Pyrophosphate, which may be deficient in advanced renal failure, is a potent inhibitor of vascular calcification. To explore its use as a potential therapeutic, we injected exogenous pyrophosphate subcutaneously or intraperitoneally in normal rats and found that their plasma pyrophosphate concentrations peaked within 15 min. There was a single exponential decay with a half-life of 33 min. The kinetics were indistinguishable between the two routes of administration or in anephric rats. The effect of daily intraperitoneal pyrophosphate injections on uremic vascular calcification was then tested in rats fed a high-phosphate diet containing adenine for 28 days to induce uremia. Although the incidence of aortic calcification varied and was not altered by pyrophosphate, the calcium content of calcified aortas was significantly reduced by 70%. Studies were repeated in uremic rats given calcitriol to produce more consistent aortic calcification and treated with sodium pyrophosphate delivered intraperitoneally in a larger volume of glucose-containing solution to prolong plasma pyrophosphate levels. This maneuver significantly reduced both the incidence and amount of calcification. Quantitative histomorphometry of bone samples after double-labeling with calcein indicated that there was no effect of pyrophosphate on the rates of bone formation or mineralization. Thus, exogenous pyrophosphate can inhibit uremic vascular calcification without producing adverse effects on bone. PMID:21124302

  20. Modulation of calcium oxalate dihydrate growth by selective crystal-face binding of phosphorylated osteopontin and polyaspartate peptide showing occlusion by sectoral (compositional) zoning.

    PubMed

    Chien, Yung-Ching; Masica, David L; Gray, Jeffrey J; Nguyen, Sarah; Vali, Hojatollah; McKee, Marc D

    2009-08-28

    Calcium oxalate dihydrate (COD) mineral and the urinary protein osteopontin/uropontin (OPN) are commonly found in kidney stones. To investigate the effects of OPN on COD growth, COD crystals were grown with phosphorylated OPN or a polyaspartic acid-rich peptide of OPN (DDLDDDDD, poly-Asp(86-93)). Crystals grown with OPN showed increased dimensions of the {110} prismatic faces attributable to selective inhibition at this crystallographic face. At high concentrations of OPN, elongated crystals with dominant {110} faces were produced, often with intergrown, interpenetrating twin crystals. Poly-Asp(86-93) dose-dependently elongated crystal morphology along the {110} faces in a manner similar to OPN. In crystal growth studies using fluorescently tagged poly-Asp(86-93) followed by imaging of crystal interiors using confocal microscopy, sectoral (compositional) zoning in COD was observed resulting from selective binding and incorporation (occlusion) of peptide exclusively into {110} crystal sectors. Computational modeling of poly-Asp(86-93) adsorption to COD {110} and {101} surfaces also suggests increased stabilization of the COD {110} surface and negligible change to the natively stable {101} surface. Ultrastructural, colloidal-gold immunolocalization of OPN by transmission electron microscopy in human stones confirmed an intracrystalline distribution of OPN. In summary, OPN and its poly-Asp(86-93) sequence similarly affect COD mineral growth; the {110} crystallographic faces become enhanced and dominant attributable to {110} face inhibition by the protein/peptide, and peptides can incorporate into the mineral phase. We, thus, conclude that the poly-Asp(86-93) domain is central to the OPN ability to interact with the {110} faces of COD, where it binds to inhibit crystal growth with subsequent intracrystalline incorporation (occlusion).

  1. Modulation of Calcium Oxalate Dihydrate Growth by Selective Crystal-face Binding of Phosphorylated Osteopontin and Polyaspartate Peptide Showing Occlusion by Sectoral (Compositional) Zoning*

    PubMed Central

    Chien, Yung-Ching; Masica, David L.; Gray, Jeffrey J.; Nguyen, Sarah; Vali, Hojatollah; McKee, Marc D.

    2009-01-01

    Calcium oxalate dihydrate (COD) mineral and the urinary protein osteopontin/uropontin (OPN) are commonly found in kidney stones. To investigate the effects of OPN on COD growth, COD crystals were grown with phosphorylated OPN or a polyaspartic acid-rich peptide of OPN (DDLDDDDD, poly-Asp86–93). Crystals grown with OPN showed increased dimensions of the {110} prismatic faces attributable to selective inhibition at this crystallographic face. At high concentrations of OPN, elongated crystals with dominant {110} faces were produced, often with intergrown, interpenetrating twin crystals. Poly-Asp86–93 dose-dependently elongated crystal morphology along the {110} faces in a manner similar to OPN. In crystal growth studies using fluorescently tagged poly-Asp86–93 followed by imaging of crystal interiors using confocal microscopy, sectoral (compositional) zoning in COD was observed resulting from selective binding and incorporation (occlusion) of peptide exclusively into {110} crystal sectors. Computational modeling of poly-Asp86–93 adsorption to COD {110} and {101} surfaces also suggests increased stabilization of the COD {110} surface and negligible change to the natively stable {101} surface. Ultrastructural, colloidal-gold immunolocalization of OPN by transmission electron microscopy in human stones confirmed an intracrystalline distribution of OPN. In summary, OPN and its poly-Asp86–93 sequence similarly affect COD mineral growth; the {110} crystallographic faces become enhanced and dominant attributable to {110} face inhibition by the protein/peptide, and peptides can incorporate into the mineral phase. We, thus, conclude that the poly-Asp86–93 domain is central to the OPN ability to interact with the {110} faces of COD, where it binds to inhibit crystal growth with subsequent intracrystalline incorporation (occlusion). PMID:19581305

  2. Anti-proliferative activity of L-651,582 correlates with calcium-mediated regulation of nucleotide metabolism at phosphoribosyl pyrophosphate synthetase

    SciTech Connect

    Hupe, D.J.; Behrens, N.D.; Boltz, R. )

    1990-09-01

    L-651,582, 5-amino-(4-(4-chlorobenzoyl)-3,5-dichlorobenzyl)-1, 2,3-triazole-4-carboxamide, is an antiproliferative and antiparasitic agent which inhibits nucleotide metabolism in mammalian cells. The drug equivalently inhibited 3H-hypoxanthine, 14C-adenine, and 14C-formate incorporation into nucleotide pools in Madin-Darby bovine kidney (MDBK) cells, suggesting depletion of the supply of phosphoribosyl pyrophosphate, (PRPP), required for each of these independent pathways. Inhibition of nucleotide metabolism correlated with inhibition of proliferation for three cell types with differing sensitivities toward the drug. L-651,582 inhibited incorporation of 3H-hypoxanthine into nucleotide pools with either glucose, uridine, or ribose as carbon source suggesting a block at PRPP synthetase, rather than a block in a pathway supplying ribose-5-phosphate. PRPP synthetase was not inhibited directly by the compound, indicating regulation of the enzyme in intact cells. Drug treatment did not kill cells but reduced the fraction of cells in S and G2/M while increasing the population in G1. Inhibition of uptake of 45Ca was demonstrated at concentrations identical to those required for inhibition of nucleotide metabolism or proliferation. Inhibition of cellular PRPP biosynthesis rates were also observed using EGTA to lower calcium levels. These data suggest a previously unrecognized link between calcium entry, the regulation of nucleotide biosynthesis at PRPP synthetase, and the rate of proliferation of mammalian cells.

  3. Crystallization of calcium sulfate dihydrate under simulated conditions of phosphoric acid production in the presence of aluminum and magnesium ions

    NASA Astrophysics Data System (ADS)

    Rashad, M. M.; Mahmoud, M. H. H.; Ibrahim, I. A.; Abdel-Aal, E. A.

    2004-06-01

    The effect of Al 3+ and Mg 2+ ions, as additives, on the crystallization of gypsum was studied under simulated conditions of the phosphoric acid production. Calcium hydrogen phosphate and sulfuric acid were mixed with dilute phosphoric acid at 80°C, and the turbidity of the reaction mixture was measured at different time periods to calculate the induction time of gypsum crystals formation. Addition of Al 3+ ions up to 2% decreased the induction time and increased the growth efficiency while addition of Mg 2+ increased the induction time and decreased the growth efficiency compared with in absence of additives. Interestingly, the crystals mean and median diameters were found to increase in the presence of Al 3+ and decrease in the presence of Mg 2+. The surface energy increased with Al 3+ and decreased with Mg 2+ compared to the baseline (without additives). Gypsum morphology changed from needle-like type in absence of additives to thick-rhombic in the presence of Al 3+ ions.

  4. Study of polymeric additive effect on calcium oxalate dihydrate crystal growth using real-time atomic force microscopy

    NASA Astrophysics Data System (ADS)

    Jung, Taesung; Kim, Jong-Nam; Kim, Woo-Sik; Kyun Choi, Chang

    2011-07-01

    Microscopic events associated with crystal growth and characterization of the growth hillocks on the (1 0 0) and (1 0 1) faces of COD were examined by atomic force microscopy. The (1 0 0) and (1 0 1) faces of COD developed elliptical and triangular hillocks and pits, respectively. Each face exhibited hillocks with step sites that can be assigned to specific crystal planes, enabling direct determination of the growth rates along specific crystallographic directions. The addition of macromolecules with anionic side chains, poly- L-aspartate, poly- L-glutamate, and polyacrylate resulted in inhibition of growth on the hillock step planes. The magnitude of their effect depended on the macromolecule structures and identity of the step site. The isotropic shape of the COD hillocks mimicked the shape of the resulting macroscopic COD crystals based on step-specific binding of the macromolecules to the COD crystal, with stronger step pinning along the [0 1 0] direction than in the [0 0 1] direction. Electrostatic matching between the crystal faces and additives according to the ionic array of calcium oxalate in the COD structure was found to be responsible for the preferential binding of the macromolecules to terraces.

  5. The detection of calcium pyrophosphate crystals in the synovial fluid of patients with rheumatoid arthritis using the cytospin technique: prevalence and clinical correlation.

    PubMed

    Theiler, Georg; Quehenberger, Franz; Rainer, Franz; Neubauer, Manfred; Stettin, Mariana; Robier, Christoph

    2014-01-01

    There are only a few studies dealing with the detection and clinical impact of calcium pyrophosphate (CPPD) crystals in patients with rheumatoid arthritis (RA) published to date. In particular, data determined by the cytospin technique, which is an effective tool to enhance the crystal detection rate, are lacking. The objectives of this study were to determine the prevalence of CPPD crystals in the synovial fluid (SF) of patients with RA and to investigate whether the detection of CPPD crystals is correlated with demographic, clinical and serological features. We examined 113 consecutive SF samples of patients with RA, obtained from therapeutic arthrocentesis of knee joints. After cytocentrifugation, the sediments were examined by polarized microscopy for the occurrence of CPPD crystals. Demographic, clinical and serological data, acquired from the medical records, were compared between crystal-positive and crystal-negative subjects. CPPD crystals were observed in 20 of the 113 cases, representing 17.7%. CPPD-positive and CPPD-negative subjects did not differ significantly in sex, duration of disease, Steinbrocker radiologic stage, disease activity score 28, as well as serum rheumatoid factor and anti-CCP positivity. Patients positively tested for CPPD crystals had a significantly higher age than CPPD-negative patients (p < 0.0001). An age-independent association of long-time treatment with diuretics and CPPD crystal formation was not found. In conclusion, demographic, clinical and serological characteristics of patients with RA were not associated with the occurrence of CPPD crystals. Age was the only significant influencing factor on CPPD crystal formation in patients with RA.

  6. A 3-day delay in synovial fluid crystal identification did not hinder the reliable detection of monosodium urate and calcium pyrophosphate crystals.

    PubMed

    Tausche, Anne-Kathrin; Gehrisch, Siegmund; Panzner, Ines; Winzer, Maria; Range, Ursula; Bornstein, Stefan R; Siegert, Gabriele; Wunderlich, Carsten; Aringer, Martin

    2013-08-01

    Arthrocentesis is an essential emergency step in managing patients with acute arthritis. To identify a bacterial infection, Gram staining is performed promptly. However, crystal analysis may not be immediately performed in many facilities. Being considered not to be stable over time, synovial fluid (SF) is sometimes discarded instead of being stored for crystal identification. The aim of this study was to assess the detectability of monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals in SF over a period of 3 days. Consecutive SF samples from 75 joints were analyzed for MSU, CPP crystals, and pH. Two independent observers evaluated the samples by regular light and polarization microscopy immediately after arthrocentesis and after 1, 2, and 3 days at room temperature or at 4°C. Of 75 samples, 27 contained crystals (16 MSU, 6 CPP, 5 both); semiquantitative counts of both MSU and CPP crystals did not change significantly after 3 days. There was no new formation of crystals in any of the crystal-negative samples, which was independent of the storage temperature. Synovial fluid pH was not predictive of crystals and did not change over time. Although immediate workup for microbiology, including Gram stain and culture, is indispensable and well established, crystal analysis may at times not be immediately performed. Our study suggests that when crystal identification cannot be done immediately, it can be safely performed up to 3 days after arthrocentesis when SF is stored at 4°C or even at stable room temperature (20°C).

  7. Bone pyrophosphate in uremia and its association with extraosseous calcification.

    PubMed Central

    Alfrey, A C; Solomons, C C

    1976-01-01

    The mean bone pyrophosphate was 0.360 +/- 0.15 mg/g in 8 controls and 1.22 +/- 1.39 mg/g bone in 27 uremic patients (P less than 0.0025). 13 of the 27 uremic patients had bone pyrophosphate levels greater than 2 SD above control values. The ash content of uremic bones with increased pyrophosphate levels (group II) was 56 +/- 9% as compared to 64 +/- 2% in control bones (P less than 0.01) and 60 +/- 7% in uremic bones having normal pyrophosphate levels (P less than 0.1) (group I). The magnesium content of bones in group II was 338 +/- 47 as compared to 211 +/- 13 (P less than 0.0005) in the controls and 294 +/- 73 mmol/kg ash (P less than 0.05) in group I. In group II, but not group I, there was a significant inverse correlation between duration of dialysis and percent bone ash (r = -0.59) (P less than 0.05). A definite relationship existed between elevated bone pyrophosphate levels and soft tissue calcification. In group II the mean pulmonary calcium content was 530 +/- 459 as compared to 32 +/- 26 mmol/kg/ash in group I (P less than 0.0025). All patients with a bone pyrophosphate level greater than 1.4 mg/g bone had extensive pulmonary calcification. It is concluded that the excess bone pyrophosphate present in some uremic patients is either deposited in the apatite crystal in the transphosphorylated form or else as the magnesium salt since the pyrophosphate is resistant to pyrophosphatase and surface adsorption of pyrophosphate is not altered by the increased bone pyrophosphate levels. The excess bone pyrophosphate could disturb bone calcification mechanisms in uremic patients. The association between increased bone pyrophosphate and soft tissue calcification suggests that the disordered pyrophosphate metabolism may be important in the pathogenesis of extraosseous calcification. PMID:175092

  8. [Pyrophosphate in medicine].

    PubMed

    Goldman, Adrian; Boije af Gennis, Gustav; Xhaard, Henri; Meri, Seppo; Yli-Kauhaluoma, Jari

    2016-01-01

    In all organisms from bacteria to humans, specific hydrolases--pyrophosphatases--hydrolyse inorganic pyrophosphate to phosphate. Without this, DNA, RNA and protein synthesis stops. Pyrophosphatases are thus essential for all life. In humans, disorders in pyrophosphate metabolism cause chondrocalcinosis and hypophosphatasia. Currently, pyrophosphate analogues, e.g. alendronate, are in clinical use in osteoporosis and Paget's disease but also for e.g. complications of prostate cancer. In bacteria and protozoan parasites, membrane-bound pyrophosphatases (mPPases), which do not occur in humans, convert pyrophosphate to a proton or sodium gradient. mPPases, which are crucial for protozoan parasites, are thus promising drug targets e.g. for malaria and leishmaniasis.

  9. Alizarin red S staining as a screening test to detect calcium compounds in synovial fluid.

    PubMed

    Paul, H; Reginato, A J; Schumacher, H R

    1983-02-01

    A simple, rapid screening method using alizarin red S stain and ordinary light microscopy to detect microcrystalline or noncrystalline calcium phosphate salts was used on wet drop preparations of synovial fluids. This proved to be helpful in detecting apatite crystal clumps and small calcium pyrophosphate dihydrate (CPPD) crystals missed by polarized light. The staining was positive in 100% of synovial fluids from patients later proven to have apatite and/or CPPD deposition diseases. Apatite and CPPD crystals were commonly found together in the same fluids. In addition, some synovial fluids from patients with osteoarthritis, renal failure dialysis, rheumatoid arthritis, and gout also exhibited positive staining. The correlation of positive alizarin red S staining with radiologic evidence of osteoarthritis suggests that apatite crystals might be related to articular cartilage degeneration in different rheumatic diseases.

  10. The effect of intracrystalline and surface-bound osteopontin on the attachment of calcium oxalate dihydrate crystals to Madin-Darby canine kidney (MDCK) cells in ultrafiltered human urine.

    PubMed

    Thurgood, Lauren A; Sørensen, Esben S; Ryall, Rosemary L

    2012-04-01

    To determine the effects of intracrystalline (IC), surface-bound (SB) and combined IC + SB osteopontin (OPN) on the binding of urinary calcium oxalate dihydrate (COD) crystals to Madin-Darby canine kidney (MDCK-II) cells in ultrafiltered (UF) human urine. (14)C-oxalic acid-labelled urinary COD crystals containing IC OPN were generated in pooled UF human urine containing human milk OPN at concentrations of 0, 1.0 and 5.0 mg/L. Additional labelled crystals were nucleated from a separate sample of the same pooled UF urine, to which were later added the same amounts of protein to produce crystals with SB OPN. COD crystals with IC+SB OPN were prepared using a combination of both techniques. Control crystals were prepared in the absence of OPN. Crystals were incubated with MDCK-II cells for up to 180 min in UF urine adjusted to 8 mm Ca(2+). Binding values for individual concentrations at specific time points and overall differences between binding curves were compared using the Mann-Whitney U-test. Crystal morphology and attachment to the cells were confirmed using field emission scanning electron microscopy (FESEM). The sizes of crystals precipitated from UF urine in the presence of 0, 1 and 5 mg/L OPN were 21.9 µm, 19.3 µm and 16.5 µm, indicating that OPN had inhibited crystal growth in a dose-dependent fashion. Binding curves for control crystals were smooth, while those of the IC and IC+SB COD crystals associated with 1 and 5 mg/L OPN were bimodal, as were those of the 1 mg/L SB crystals. This suggests that OPN induces or potentiates a transient response that enables MDCK-II cells to release COD crystals after they have attached. Although OPN generally reduced the binding of urinary COD crystals to MDCK-II cells, at times it also appeared to mediate adhesion. It is possible therefore that OPN can reduce or increase crystal binding, and that our data represent the net effect of its opposing inhibitory or promotory properties. In UF urine, OPN inhibits the growth of

  11. Development of optically transparent water oxidation catalysts using manganese pyrophosphate compounds.

    PubMed

    Takashima, Toshihiro; Hotori, Yuki; Irie, Hiroshi

    2015-11-01

    One challenge in artificial photosynthetic systems is the development of active oxygen evolution catalysts composed of abundant elements. The oxygen evolution activities of manganese pyrophosphate compounds were examined in electrochemical and photochemical experiments. Electrocatalysis using calcium-manganese pyrophosphate exhibited good catalytic ability under neutral pH and an oxygen evolution reaction was driven with a small overpotential (η<100 mV). UV-vis diffuse reflectance measurements revealed that manganese pyrophosphates exhibit weak absorption in the visible light region while commonly used oxygen evolution catalysts exhibit intense absorption. Therefore, the efficient light absorption of a photocatalyst was retained even after surface modification with a manganese pyrophosphate, and photochemical oxygen evolution was achieved by using magnesium ferrite modified with manganese pyrophosphate nanoparticles under the illumination of visible light at wavelength of over 420 nm. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Calcium Input Potentiates the Transforming Growth Factor (TGF)-β1-dependent Signaling to Promote the Export of Inorganic Pyrophosphate by Articular Chondrocyte*

    PubMed Central

    Cailotto, Frederic; Reboul, Pascal; Sebillaud, Sylvie; Netter, Patrick; Jouzeau, Jean-Yves; Bianchi, Arnaud

    2011-01-01

    Transforming growth factor (TGF)-β1 stimulates extracellular PPi (ePPi) generation and promotes chondrocalcinosis, which also occurs secondary to hyperparathyroidism-induced hypercalcemia. We previously demonstrated that ANK was up-regulated by TGF-β1 activation of ERK1/2 and Ca2+-dependent protein kinase C (PKCα). Thus, we investigated mechanisms by which calcium could affect ePPi metabolism, especially its main regulating proteins ANK and PC-1 (plasma cell membrane glycoprotein-1). We stimulated articular chondrocytes with TGF-β1 under extracellular (eCa2+) or cytosolic Ca2+ (cCa2+) modulations. We studied ANK, PC-1 expression (quantitative RT-PCR, Western blotting), ePPi levels (radiometric assay), and cCa2+ input (fluorescent probe). Voltage-operated Ca2+-channels (VOC) and signaling pathways involved were investigated with selective inhibitors. Finally, Ank promoter activity was evaluated (gene reporter). TGF-β1 elevated cCa2+ and ePPi levels (by up-regulating Ank and PC-1 mRNA/proteins) in an eCa2+ dose-dependent manner. TGF-β1 effects were suppressed by cCa2+ chelation or L- and T-VOC blockade while being mostly reproduced by ionomycin. In the same experimental conditions, the activation of Ras, the phosphorylation of ERK1/2 and PKCα, and the stimulation of Ank promoter activity were affected similarly. Activation of SP1 (specific protein 1) and ELK-1 (Ets-like protein-1) transcription factors supported the regulatory role of Ca2+. SP1 or ELK-1 overexpression or blockade experiments demonstrated a major contribution of ELK-1, which acted synergistically with SP1 to activate Ank promoter in response to TGF-β1. TGF-β1 promotes input of eCa2+ through opening of L- and T-VOCs, to potentiate ERK1/2 and PKCα signaling cascades, resulting in an enhanced activation of Ank promoter and ePPi production in chondrocyte. PMID:21471198

  13. Calcium input potentiates the transforming growth factor (TGF)-beta1-dependent signaling to promote the export of inorganic pyrophosphate by articular chondrocyte.

    PubMed

    Cailotto, Frederic; Reboul, Pascal; Sebillaud, Sylvie; Netter, Patrick; Jouzeau, Jean-Yves; Bianchi, Arnaud

    2011-06-03

    Transforming growth factor (TGF)-β1 stimulates extracellular PP(i) (ePP(i)) generation and promotes chondrocalcinosis, which also occurs secondary to hyperparathyroidism-induced hypercalcemia. We previously demonstrated that ANK was up-regulated by TGF-β1 activation of ERK1/2 and Ca(2+)-dependent protein kinase C (PKCα). Thus, we investigated mechanisms by which calcium could affect ePP(i) metabolism, especially its main regulating proteins ANK and PC-1 (plasma cell membrane glycoprotein-1). We stimulated articular chondrocytes with TGF-β1 under extracellular (eCa(2+)) or cytosolic Ca(2+) (cCa(2+)) modulations. We studied ANK, PC-1 expression (quantitative RT-PCR, Western blotting), ePP(i) levels (radiometric assay), and cCa(2+) input (fluorescent probe). Voltage-operated Ca(2+)-channels (VOC) and signaling pathways involved were investigated with selective inhibitors. Finally, Ank promoter activity was evaluated (gene reporter). TGF-β1 elevated cCa(2+) and ePP(i) levels (by up-regulating Ank and PC-1 mRNA/proteins) in an eCa(2+) dose-dependent manner. TGF-β1 effects were suppressed by cCa(2+) chelation or L- and T-VOC blockade while being mostly reproduced by ionomycin. In the same experimental conditions, the activation of Ras, the phosphorylation of ERK1/2 and PKCα, and the stimulation of Ank promoter activity were affected similarly. Activation of SP1 (specific protein 1) and ELK-1 (Ets-like protein-1) transcription factors supported the regulatory role of Ca(2+). SP1 or ELK-1 overexpression or blockade experiments demonstrated a major contribution of ELK-1, which acted synergistically with SP1 to activate Ank promoter in response to TGF-β1. TGF-β1 promotes input of eCa(2+) through opening of L- and T-VOCs, to potentiate ERK1/2 and PKCα signaling cascades, resulting in an enhanced activation of Ank promoter and ePP(i) production in chondrocyte.

  14. Pyrophosphate Stimulates Differentiation, Matrix Gene Expression and Alkaline Phosphatase Activity in Osteoblasts

    PubMed Central

    Pujari-Palmer, Michael; Pujari-Palmer, Shiuli; Lu, Xi; Lind, Thomas; Melhus, Håkan; Engstrand, Thomas; Karlsson-Ott, Marjam; Engqvist, Hakan

    2016-01-01

    Pyrophosphate is a potent mitogen, capable of stimulating proliferation in multiple cell types, and a critical participant in bone mineralization. Pyrophosphate can also affect the resorption rate and bioactivity of orthopedic ceramics. The present study investigated whether calcium pyrophosphate affected proliferation, differentiation and gene expression in early (MC3T3 pre-osteoblast) and late stage (SAOS-2 osteosarcoma) osteoblasts. Pyrophosphate stimulated peak alkaline phosphatase activity by 50% and 150% at 100μM and 0.1μM in MC3T3, and by 40% in SAOS-2. The expression of differentiation markers collagen 1 (COL1), alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) were increased by an average of 1.5, 2, 2 and 3 fold, by high concentrations of sodium pyrophosphate (100μM) after 7 days of exposure in MC3T3. COX-2 and ANK expression did not differ significantly from controls in either treatment group. Though both high and low concentrations of pyrophosphate stimulate ALP activity, only high concentrations (100μM) stimulated osteogenic gene expression. Pyrophosphate did not affect proliferation in either cell type. The results of this study confirm that chronic exposure to pyrophosphate exerts a physiological effect upon osteoblast differentiation and ALP activity, specifically by stimulating osteoblast differentiation markers and extracellular matrix gene expression. PMID:27701417

  15. Arthritis

    MedlinePlus

    ... joints Infection, most often by bacteria or virus Crystals such as uric acid or calcium pyrophosphate dihydrate ... common types of inflammatory arthritis include: Ankylosing ... calcium pyrophosphate deposition disease Juvenile rheumatoid ...

  16. Permanent stained preparations of synovial fluid for detection of calcium compounds using alizarin red S.

    PubMed

    Lazcano, O; Bilbao, J; Beissner, R S; Vandiver, M; Li, C Y

    1992-01-01

    Permanent preparations of air dried synovial fluids were prepared by staining calcium compounds with alizarin red S stain; each slide was coverslipped with Permount. Variables studied were: (a) concentration of the solution of alizarin red S, (b) pH of staining solution, (c) time of incubation in staining solution and aqueous and ethanolic content of staining solution. The staining effect of each solution was tested on calcium pyrophosphate dihydrate, calcium oxalate, apatite and monosodium urate (MSU). Of all the solutions, best results were obtained with 0.25% alizarin red S in 50% ethanol at pH 7.0 for 30 min. With this solution, the calcium-containing compounds were well stained. MSU did not stain and still preserved negative birefringence on polarization. Fixation of smears with ethanol served a double purpose: It fixed the slides without dissolving or removing MSU or the calcium compounds, yet it did dissolve five corticosteroids commonly used for intra-articular injection which may interfere with interpretation of compensated polarized light microscopy of synovial fluids.

  17. 21 CFR 182.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium pyrophosphate. 182.6787 Section 182.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Conditions of use. This substance...

  18. 21 CFR 182.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium pyrophosphate. 182.6787 Section 182.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Conditions of use. This substance...

  19. 21 CFR 582.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Sodium pyrophosphate. 582.6787 Section 582.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Condition of use. This substance...

  20. 21 CFR 582.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium pyrophosphate. 582.6787 Section 582.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Condition of use. This substance...

  1. 21 CFR 182.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Sodium pyrophosphate. 182.6787 Section 182.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Conditions of use. This substance...

  2. 21 CFR 582.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium pyrophosphate. 582.6787 Section 582.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Condition of use. This substance...

  3. 21 CFR 182.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Sodium pyrophosphate. 182.6787 Section 182.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Conditions of use. This substance...

  4. 21 CFR 582.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium pyrophosphate. 582.6787 Section 582.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Condition of use. This substance...

  5. 21 CFR 582.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium pyrophosphate. 582.6787 Section 582.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Sodium pyrophosphate. (a) Product. Sodium pyrophosphate. (b) Condition of use. This substance...

  6. 21 CFR 582.5304 - Ferric pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ferric pyrophosphate. 582.5304 Section 582.5304 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5304 Ferric pyrophosphate. (a) Product. Ferric pyrophosphate. (b) Conditions of use. This...

  7. 21 CFR 582.5304 - Ferric pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ferric pyrophosphate. 582.5304 Section 582.5304 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5304 Ferric pyrophosphate. (a) Product. Ferric pyrophosphate. (b) Conditions of use. This...

  8. 21 CFR 582.5304 - Ferric pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Ferric pyrophosphate. 582.5304 Section 582.5304 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5304 Ferric pyrophosphate. (a) Product. Ferric pyrophosphate. (b) Conditions of use. This...

  9. 21 CFR 582.5304 - Ferric pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Ferric pyrophosphate. 582.5304 Section 582.5304 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Supplements 1 § 582.5304 Ferric pyrophosphate. (a) Product. Ferric pyrophosphate. (b) Conditions of use. This...

  10. Calcium

    MedlinePlus

    ... You'll also find calcium in broccoli and dark green, leafy vegetables (especially collard and turnip greens, ... can enjoy good sources of calcium such as dark green, leafy vegetables, broccoli, chickpeas, and calcium-fortified ...

  11. The different phases in the precipitation of dicalcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Ferreira, A.; Oliveira, C.; Rocha, F.

    2003-05-01

    The precipitation of dicalcium phosphate dihydrate, brushite, by mixing a calcium hydroxide suspension and an orthophosphoric acid solution in equimolar quantities, has been investigated in a batch system at 25°C. The concentration of calcium hydroxide and orthophosphoric acid, before mixing, ranged from 50 to 300 mmol dm -3. The phase first precipitated is Ca 5OH(PO 4) 3, hydroxyapatite. The precipitation process of brushite is divided into five stages and is similar for all initial experimental conditions. The extension of each stage varies with the initial reagents' concentrations. These stages are discussed individually as a function of pH and reagents' concentrations. The precipitate was analysed by scanning electron microscopy and X-ray diffraction. The solubility of brushite was determined at 25°C, 30°C and 35°C, and in the pH range 4.5-8.

  12. Inorganic pyrophosphate generation by transforming growth factor-beta-1 is mainly dependent on ANK induction by Ras/Raf-1/extracellular signal-regulated kinase pathways in chondrocytes.

    PubMed

    Cailotto, Frederic; Bianchi, Arnaud; Sebillaud, Sylvie; Venkatesan, Narayanan; Moulin, David; Jouzeau, Jean-Yves; Netter, Patrick

    2007-01-01

    ANK is a multipass transmembrane protein transporter thought to play a role in the export of intracellular inorganic pyrophosphate and so to contribute to the pathophysiology of chondrocalcinosis. As transforming growth factor-beta-1 (TGF-beta1) was shown to favor calcium pyrophosphate dihydrate deposition, we investigated the contribution of ANK to the production of extracellular inorganic pyrophosphate (ePPi) by chondrocytes and the signaling pathways involved in the regulation of Ank expression by TGF-beta1. Chondrocytes were exposed to 10 ng/mL of TGF-beta1, and Ank expression was measured by quantitative polymerase chain reaction and Western blot. ePPi was quantified in cell supernatants. RNA silencing was used to define the respective roles of Ank and PC-1 in TGF-beta1-induced ePPi generation. Finally, selective kinase inhibitors and dominant-negative/overexpression plasmid strategies were used to explore the contribution of several signaling pathways to Ank induction by TGF-beta1. TGF-beta1 strongly increased Ank expression at the mRNA and protein levels, as well as ePPi production. Using small interfering RNA technology, we showed that Ank contributed approximately 60% and PC-1 nearly 20% to TGF-beta1-induced ePPi generation. Induction of Ank by TGF-beta1 required activation of the extracellular signal-regulated kinase (ERK) pathway but not of p38-mitogen-activated protein kinase or of protein kinase A. In line with the general protein kinase C (PKC) inhibitor calphostin C, Gö6976 (a Ca2+-dependent PKC inhibitor) diminished TGF-beta1-induced Ank expression by 60%, whereas a 10% inhibition was observed with rottlerin (a PKCdelta inhibitor). These data suggest a regulatory role for calcium in TGF-beta1-induced Ank expression. Finally, we demonstrated that the stimulatory effect of TGF-beta1 on Ank expression was inhibited by the suppression of the Ras/Raf-1 pathway, while being enhanced by their constitutive activation. Transient overexpression of Smad 7, an

  13. Calcium

    MedlinePlus

    ... in luck if you like sardines and canned salmon with bones. Almond milk. previous continue Working Calcium ... drinks, and cereals. Other Considerations for Building Bones Vitamin D is essential for calcium absorption, so it's ...

  14. Calcium

    MedlinePlus

    ... such as canned sardines and salmon Calcium-enriched foods such as breakfast cereals, fruit juices, soy and rice drinks, and tofu. Check the product labels. The exact amount of calcium you need depends on your age and other factors. Growing children and teenagers need more calcium than ...

  15. Enrofloxacin hydro-chloride dihydrate.

    PubMed

    Miranda-Calderón, Jorge E; Gutiérrez, Lilia; Flores-Alamo, Marcos; García-Gutiérrez, Ponciano; Sumano, Héctor

    2014-04-01

    The asymmetric unit of the title compound, C19H23FN3O3 (+)·Cl(-)·2H2O [systematic name: 4-(3-carb-oxy-1-cyclo-propyl-6-fluoro-4-oxo-1,4-di-hydro-quin-o-lin-7-yl)-1-ethyl-piperazin-1-ium chloride dihydrate], consists of two independent monocations of the protonated enrofloxacin, two chloride anions and four water mol-ecules. In the cations, the piperazinium rings adopt chair conformations and the dihedral angles between the cyclo-propyl ring and the 10-membered quinoline ring system are 56.55 (2) and 51.11 (2)°. An intra-molecular O-H⋯O hydrogen bond is observed in each cation. In the crystal, the components are connected via O-H⋯Cl, N-H⋯Cl and O-H⋯O hydrogen bonds, and a π-π inter-action between the benzene rings [centroid-centroid distance = 3.6726 (13) Å], resulting in a three-dimensional array.

  16. Feasibility of a tetracycline-binding method for detecting synovial fluid basic calcium phosphate crystals.

    PubMed

    Rosenthal, Ann K; Fahey, Mark; Gohr, Claudia; Burner, Todd; Konon, Irina; Daft, Laureen; Mattson, Eric; Hirschmugl, Carol; Ryan, Lawrence M; Simkin, Peter

    2008-10-01

    Basic calcium phosphate (BCP) crystals are common components of osteoarthritis (OA) synovial fluid. Progress in understanding the role of these bioactive particles in clinical OA has been hampered by difficulties in their identification. Tetracyclines stain calcium phosphate mineral in bone. The aim of this study was to investigate whether tetracycline staining might be an additional or alternative method for identifying BCP crystals in synovial fluid. A drop of oxytetracycline was mixed with a drop of fluid containing synthetic or native BCP, calcium pyrophosphate dihydrate (CPPD), or monosodium urate (MSU) crystals and placed on a microscope slide. Stained and unstained crystals were examined by light microscopy, with and without a portable broad-spectrum ultraviolet (UV) pen light. A small set of characterized synovial fluid samples were compared by staining with alizarin red S and oxytetracycline. Synthetic BCP crystals in synovial fluid were quantified fluorimetrically using oxytetracycline. After oxytetracycline staining, synthetic and native BCP crystals appeared as fluorescent amorphous aggregates under UV light. Oxytetracycline did not stain CPPD or MSU crystals or other particulates. Oxytetracycline staining had fewer false-positive test results than did alizarin red S staining and could provide estimates of the quantities of synthetic BCP crystals in synovial fluid. With further validation, oxytetracycline staining may prove to be a useful adjunct or alternative to currently available methods for identifying BCP crystals in synovial fluid.

  17. 21 CFR 182.6787 - Sodium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Sodium pyrophosphate. 182.6787 Section 182.6787 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Sequestrants 1 § 182.6787 Sodium pyrophosphate. (a) Product....

  18. PREPARATION OF ALKYL PYROPHOSPHATE EXTRACTANTS

    DOEpatents

    Levine, C.A.; Skiens, W.E.; Moore, G.R.

    1960-08-01

    A process for providing superior solvent extractants for metal recovery processes is given wherein the extractant comprises an alkyl pyrophosphoric acid ester dissolved in an organic solvent diluent. Finely divided solid P/sub 2/O/ sub 5/ is slurried in an organic solvent-diluent selected from organic solvents such as kerosene, benzene, chlorobenzene, toluene, etc. An alcohol selected from the higher alcohols having 4 to 17 carbon atoms. e.g.. hexanol-1. heptanol-3, octanol-1. 2.6-dimethyl-heptanol-4, and decanol-1, is rapidly added to the P/sub 2/O/sub 5/ slurry in the amount of about 2 moles of alcohol to 1 mole of P/sub 2/ O/sub 5/. The temperature is maintained below about 110 deg C during the course of the P/sub 2/O/sub 5/-alcohol reaction. An alkyl pyrophosphate extractant compound is formed as a consequence of the reaction process. The alkyl pyrophosphate solvent-diluent extractant phase is useful in solvent extraction metal recovery processes.

  19. The Thiamin Pyrophosphate-Motif

    NASA Technical Reports Server (NTRS)

    Dominiak, Paulina M.; Ciszak, Ewa M.

    2003-01-01

    Using databases the authors have identified a common thiamin pyrophosphate (TPP)-motif in the family of functionally diverse TPP-dependent enzymes. This common motif consists of multimeric organization of subunits, two catalytic centers, common amino acid sequence, and specific contacts to provide a flip-flop, or alternate site, mechanism of action. Each catalytic center [PP:PYR] is formed at the interface of the PP-domain binding the magnesium ion, pyrophosphate and aminopyrimidine ring of TPP, and the PYR-domain binding the aminopyrimidine ring of that cofactor. A pair of these catalytic centers constitutes the catalytic core [PP:PYR]* within these enzymes. Analysis of the structural elements of this catalytic core reveals novel definition of the common amino acid sequences, which are GX@&(G)@XXGQ, and GDGX25-30 within the PP- domain, and the E&(G)@XXG@ within the PYR-domain, where Q, corresponds to a hydrophobic amino acid. This TPP-motif provides a novel tool for annotation of TPP-dependent enzymes useful in advancing functional proteomics.

  20. The Thiamin Pyrophosphate-Motif

    NASA Technical Reports Server (NTRS)

    Dominiak, Paulina M.; Ciszak, Ewa M.

    2003-01-01

    Using databases the authors have identified a common thiamin pyrophosphate (TPP)-motif in the family of functionally diverse TPP-dependent enzymes. This common motif consists of multimeric organization of subunits, two catalytic centers, common amino acid sequence, and specific contacts to provide a flip-flop, or alternate site, mechanism of action. Each catalytic center [PP:PYR] is formed at the interface of the PP-domain binding the magnesium ion, pyrophosphate and aminopyrimidine ring of TPP, and the PYR-domain binding the aminopyrimidine ring of that cofactor. A pair of these catalytic centers constitutes the catalytic core [PP:PYR]* within these enzymes. Analysis of the structural elements of this catalytic core reveals novel definition of the common amino acid sequences, which are GX@&(G)@XXGQ, and GDGX25-30 within the PP- domain, and the E&(G)@XXG@ within the PYR-domain, where Q, corresponds to a hydrophobic amino acid. This TPP-motif provides a novel tool for annotation of TPP-dependent enzymes useful in advancing functional proteomics.

  1. The Thiamin Pyrophosphate-Motif

    NASA Technical Reports Server (NTRS)

    Dominiak, P.; Ciszak, E.

    2003-01-01

    Using databases the authors have identified a common thiamin pyrophosphate (TPP)-motif in the family of functionally diverse TPP-dependent enzymes. This common motif consists of multimeric organization of subunits and two catalytic centers. Each catalytic center (PP:PYR) is formed at the interface of the PP-domain binding the magnesium ion, pyrophosphate and amhopyrimidine ring of TPP, and the PYR-domain binding the aminopyrimidine ring of that cofactor. A pair of these catalytic centers constitutes the catalytic core (PP:PYR)(sub 2) within these enzymes. Analysis of the structural elements of this catalytic core reveals novel definition of the common amino acid sequences, which are GXPhiX(sub 4)(G)PhiXXGQ and GDGX(sub 25-30)NN in the PP-domain, and the EX(sub 4)(G)PhiXXGPhi in the PYR-domain, where Phi corresponds to a hydrophobic amino acid. This TPP-motif provides a novel tool for annotation of TPP-dependent enzymes useful in advancing functional proteomics.

  2. The Thiamin Pyrophosphate-Motif

    NASA Technical Reports Server (NTRS)

    Dominiak, P.; Ciszak, E.

    2003-01-01

    Using databases the authors have identified a common thiamin pyrophosphate (TPP)-motif in the family of functionally diverse TPP-dependent enzymes. This common motif consists of multimeric organization of subunits and two catalytic centers. Each catalytic center (PP:PYR) is formed at the interface of the PP-domain binding the magnesium ion, pyrophosphate and amhopyrimidine ring of TPP, and the PYR-domain binding the aminopyrimidine ring of that cofactor. A pair of these catalytic centers constitutes the catalytic core (PP:PYR)(sub 2) within these enzymes. Analysis of the structural elements of this catalytic core reveals novel definition of the common amino acid sequences, which are GXPhiX(sub 4)(G)PhiXXGQ and GDGX(sub 25-30)NN in the PP-domain, and the EX(sub 4)(G)PhiXXGPhi in the PYR-domain, where Phi corresponds to a hydrophobic amino acid. This TPP-motif provides a novel tool for annotation of TPP-dependent enzymes useful in advancing functional proteomics.

  3. Dehydration behavior of eprosartan mesylate dihydrate.

    PubMed

    Sheng, J; Venkatesh, G M; Duddu, S P; Grant, D J

    1999-10-01

    Eprosartan mesylate (SKF 108566-J; EM) is an antihypertensive agent approved for marketing in the USA. EM dihydrate was prepared by three methods, one of which included suspending the anhydrous drug in an aqueous solution of 1.0 M methanesulfonic acid to form a slurry, followed by filtration. The dehydration kinetics of EM dihydrate were derived by analyzing the fit of the isothermal thermogravimetric analytical (TGA) data to numerous kinetic models. EM dihydrate undergoes dehydration in two distinct steps, each involving the loss of 1 mol of water at 25-70 degrees C and 70-120 degrees C, respectively. Recrystallization of EM occurs at approximately 120-140 degrees C after dehydration to the anhydrous phase. This explanation is supported by variable temperature powder X-ray diffractometry. The mechanism of the dehydration reaction is complex, the dependence of the reaction rate on temperature varying as a function of the particles size. For the dihydrate of sieve fraction <125 microm, the kinetics of the first and second dehydration steps are consistent with the Avrami-Erofeev equation (A3, n = 1/3) over the temperature range studied, corresponding to three-dimensional growth of nuclei. In contrast, for the 125-180-microm and 180-250-microm sieve fractions, the kinetics are best described by the two-dimensional phase boundary reaction (R2) at a lower dehydration temperature (i.e., 28.3 degrees C), and by the Avrami-Erofeev equation (A3, n = 1/3) at a higher dehydration temperature (i.e., 93.7 degrees C). The activation energies (15-40 kcal/mol) and frequency factors of the dehydration of EM dihydrate were determined both by Arrhenius plots of the isothermal rates determined by TGA and by Kissinger plots of the nonisothermal differential scanning calorimetric data. Hot stage microscopy of single crystals of EM dihydrate showed random nucleation at the surface and dehydration with the growth of microcrystals along the needle a axis. Cerius(2) molecular modeling

  4. Farnesyl pyrophosphate synthetase. Mechanistic studies of the 1'-4 coupling reaction with 2-fluorogeranyl pyrophosphate.

    PubMed

    Poulter, C D; Argyle, J C; Mash, E A

    1978-10-25

    The mechanism of the 1'-4 coupling reaction between isopentenyl pyrophosphate and geranyl pyrophosphate catalyzed by farnesyl pyrophosphate synthetase from porcine liver was studied with the allylic substrate analogue 2-fluorogeranyl pyrophosphate. 2-Fluorogeranyl pyrophosphate is an alternate substrate for the enzyme, yielding 6-fluorofarnesyl pyrophosphate upon condensation with isopentenyl pyrophosphate. The Michaelis constant for the fluoroanalogue, Km = 1.1 micron, is similar to that measured for geranyl pyrophosphate, Km = 0.7 micron. However, the rate of condensation with the fluoroanalogue was only 8.4 X 10(-4) that of the normal reaction. A similar rate of depression (4.4 X 10(-3)) was found for solvolysis of geranyl methanesulfonate and the corresponding 2-fluoro derivative, reactions known to proceed via cationic intermediates. In contrast, displacement of chlorine from geranyl chloride and 2-fluorogeranyl chloride by cyanide showed a small (2-fold) rate enhancement for the fluoro compound. Finally, 2-fluorogeranyl pyrophosphate is a competitive inhibitor against geranyl pyrophosphate. These data are interpreted in terms of an ionization-condensation-elimination mechanism for the 1'-4 coupling reaction.

  5. Cytocompatibility evaluation of microwave sintered biphasic calcium phosphate scaffolds synthesized using pH control.

    PubMed

    Wagner, Darcy E; Jones, Andrew D; Zhou, Huan; Bhaduri, Sarit B

    2013-04-01

    Compounds belonging to the calcium phosphate (CaP) system are known to be major constituents of bone and are bioactive to different extents in vitro and in vivo. Their chemical similarity makes them prime candidates for implants and bone tissue engineering scaffolds. CaP nanoparticles of amorphous hydroxyapatite (aHA) and dicalcium phosphate dihydrate (DCPD) were synthesized using chemical precipitation. Uniaxially pressed aHA and DCPD powders were subjected to microwave radiation to promote solid state phase transformations resulting in crystalline hydroxyapatite (HA), tricalcium phosphate (TCP) and biphasic compositions: HA/TCP and TCP/calcium pyrophosphate (CPP) and their subsequent densification. Phase composition of microwave sintered compacts was confirmed via X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). Solution pH during crystal growth was found to have a profound effect on particle morphology and post-sintered phases, despite constant sintering temperature. Cytocompatibility assessment using 7F2 cells, corresponding to adult mouse osteoblasts, on microwave and conventional, furnace sintered samples demonstrated that manufacturing method does not impact cellular viability after 24 h or proliferation over 7 days. New CaP deposition and extracellular matrix components were observed in vitro via scanning electron microscopy (SEM). Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Calcium.

    PubMed

    Williams, Robert J P

    2002-01-01

    This chapter describes the chemical and biological value of the calcium ion. In calcium chemistry, our main interest is in equilibria within static, nonflowing systems. Hence, we examined the way calcium formed precipitates and complex ions in solution. We observed thereafter its uses by humankind in a vast number of materials such as minerals, e.g., marble, concrete, mortars, which parallel the biological use in shells and bones. In complex formation, we noted that many combinations were of anion interaction with calcium for example in the uses of detergents and medicines. The rates of exchange of calcium from bound states were noted but they had little application. Calcium ions do not act as catalysts of organic reactions. In biological systems, interest is in the above chemistry, but extends to the fact that Ca2+ ions can carry information by flowing in one solution or from one solution to another through membranes. Hence, we became interested in the details of rates of calcium exchange. The fast exchange of this divalent ion from most organic binding sites has allowed it to develop as the dominant second messenger. Now the flow can be examined in vitro as calcium binds particular isolated proteins, which it activates as seen in physical mechanical changes or chemical changes and this piece-by-piece study of cells is common. Here, however, we have chosen to stress the whole circuit of Ca2+ action indicating that the cell is organized both at a basal and an activated state kinetic level by the steady state flow of the ion (see Fig. 11). Different time constants of exchange utilizing very similar binding constants lead to: 1) fast responses as in the muscle of an animal; or 2) slower change as in differentiation of an egg or seed. Many other changes of state may relate to Ca2+ steady-state levels of flow in the circuitry and here we point to two: 1) dormancy in reptiles and animals; and 2) sporulation in both bacteria and lower plants. In the other chapters of

  7. Isothermal Decomposition of Hydrogen Peroxide Dihydrate

    NASA Technical Reports Server (NTRS)

    Loeffler, M. J.; Baragiola, R. A.

    2011-01-01

    We present a new method of growing pure solid hydrogen peroxide in an ultra high vacuum environment and apply it to determine thermal stability of the dihydrate compound that forms when water and hydrogen peroxide are mixed at low temperatures. Using infrared spectroscopy and thermogravimetric analysis, we quantified the isothermal decomposition of the metastable dihydrate at 151.6 K. This decomposition occurs by fractional distillation through the preferential sublimation of water, which leads to the formation of pure hydrogen peroxide. The results imply that in an astronomical environment where condensed mixtures of H2O2 and H2O are shielded from radiolytic decomposition and warmed to temperatures where sublimation is significant, highly concentrated or even pure hydrogen peroxide may form.

  8. Isothermal Decomposition of Hydrogen Peroxide Dihydrate

    NASA Technical Reports Server (NTRS)

    Loeffler, M. J.; Baragiola, R. A.

    2011-01-01

    We present a new method of growing pure solid hydrogen peroxide in an ultra high vacuum environment and apply it to determine thermal stability of the dihydrate compound that forms when water and hydrogen peroxide are mixed at low temperatures. Using infrared spectroscopy and thermogravimetric analysis, we quantified the isothermal decomposition of the metastable dihydrate at 151.6 K. This decomposition occurs by fractional distillation through the preferential sublimation of water, which leads to the formation of pure hydrogen peroxide. The results imply that in an astronomical environment where condensed mixtures of H2O2 and H2O are shielded from radiolytic decomposition and warmed to temperatures where sublimation is significant, highly concentrated or even pure hydrogen peroxide may form.

  9. Isothermal decomposition of hydrogen peroxide dihydrate.

    PubMed

    Loeffler, M J; Baragiola, R A

    2011-06-02

    We present a new method of growing pure solid hydrogen peroxide in an ultra high vacuum environment and apply it to determine thermal stability of the dihydrate compound that forms when water and hydrogen peroxide are mixed at low temperatures. Using infrared spectroscopy and thermogravimetric analysis, we quantified the isothermal decomposition of the metastable dihydrate at 151.6 K. This decomposition occurs by fractional distillation through the preferential sublimation of water, which leads to the formation of pure hydrogen peroxide. The results imply that in an astronomical environment where condensed mixtures of H(2)O(2) and H(2)O are shielded from radiolytic decomposition and warmed to temperatures where sublimation is significant, highly concentrated or even pure hydrogen peroxide may form. © 2011 American Chemical Society

  10. ALKYL PYROPHOSPHATE METAL SOLVENT EXTRACTANTS AND PROCESS

    DOEpatents

    Long, R.L.

    1958-09-30

    A process is presented for the recovery of uranium from aqueous mineral acidic solutions by solvent extraction. The extractant is a synmmetrical dialkyl pyrophosphate in which the alkyl substituents have a chain length of from 4 to 17 carbon atoms. Mentioned as a preferred extractant is dioctyl pyrophosphate. The uranium is precipitated irom the organic extractant phase with an agent such as HF, fluoride salts. alcohol, or ammonia.

  11. Inhibition of pyruvate decarboxylase from Z. mobilis by novel analogues of thiamine pyrophosphate: investigating pyrophosphate mimics.

    PubMed

    Erixon, Karl M; Dabalos, Chester L; Leeper, Finian J

    2007-03-07

    Replacement of the thiazolium ring of thiamine pyrophosphate with a triazole gives extremely potent inhibitors of pyruvate decarboxylase from Z. mobilis, with K(I) values down to 20 pM; this system was used to explore pyrophosphate mimics and several effective analogues were discovered.

  12. The Thiamine-Pyrophosphate-Motif

    NASA Technical Reports Server (NTRS)

    Ciszak, Ewa; Dominiak, Paulina

    2004-01-01

    Thiamin pyrophosphate (TPP), a derivative of vitamin B1, is a cofactor for enzymes performing catalysis in pathways of energy production including the well known decarboxylation of a-keto acid dehydrogenases followed by transketolation. TPP-dependent enzymes constitute a structurally and functionally diverse group exhibiting multimeric subunit organization, multiple domains and two chemically equivalent catalytic centers. Annotation of functional TPP-dependcnt enzymes, therefore, has not been trivial due to low sequence similarity related to this complex organization. Our approach to analysis of structures of known TPP-dependent enzymes reveals for the first time features common to this group, which we have termed the TPP-motif. The TPP-motif consists of specific spatial arrangements of structural elements and their specific contacts to provide for a flip-flop, or alternate site, enzymatic mechanism of action. Analysis of structural elements entrained in the flip-flop action displayed by TPP-dependent enzymes reveals a novel definition of the common amino acid sequences. These sequences allow for annotation of TPP-dependent enzymes, thus advancing functional proteomics. Further details of three-dimensional structures of TPP-dependent enzymes will be discussed.

  13. The growth of dicalcium phosphate dihydrate on octacalcium phosphate at 25°C

    NASA Astrophysics Data System (ADS)

    Heughebaert, Jean-Claude; De Rooij, J. F.; Nancollas, G. H.

    1986-07-01

    The crystallization of calcium phosphate phases from metastable supersaturated solutions following seeding with well characterized octacalcium phosphate [OCP) has been studied at 25°C, using a constant composition method. At pH 6.00, and at low supersaturation (σ<1.02), OCP is grown, whereas at higher supersaturation, dicalcium phosphate dihydrate (DCPD) nucleates and grows on the OCP seed, after well-defined induction periods. The linear dependencies of the logarithm of the induction periods upon (log supersaturation) -2, is indicative of a surface nucleation process for DCPD with an effective surface energy of 7±1 mJ m -2.

  14. Is ligation the only solution to the pyrophosphate problem?

    NASA Technical Reports Server (NTRS)

    Visscher, J.; Jongbloets, R.; Schwartz, Alan W.

    1993-01-01

    Pyrophosphate linkages are easily formed during the nonenzymatic oligomerization of activated nucleotides. They often form `caps' which terminate an oligonucleotide with a 5'-5' pyrophosphate. Owing to their structural resemblance to the intermediates in enzymatic ligation reactions, it has been suggested that pyrophosphate caps might have been capable of acting as activating groups in chain elongation processes. We argue that an alternative possibility would have been the specific hydrolysis of pyrophosphates.

  15. Is ligation the only solution to the pyrophosphate problem?

    NASA Astrophysics Data System (ADS)

    Visscher, J.; Jongbloets, R.; Schwartz, Alan W.

    1993-12-01

    Pyrophosphate linkages are easily formed during the nonenzymatic oligomerization of activated nucleotides. They often form ‘caps’ which terminate an oligonucleotide with a 5'-5' pyrophosphate. Owing to their structural resemblance to the intermediates in enzymatic ligation reactions, it has been suggested that pyrophosphate caps might have been capable of acting as activating groups in chain elongation processes. We argue that an alternative possibility would have been the specific hydrolysis of pyrophosphates.

  16. A water setting tetracalcium phosphate-dicalcium phosphate dihydrate cement.

    PubMed

    Burguera, E F; Guitián, F; Chow, L C

    2004-11-01

    The development of a calcium phosphate cement, comprising tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD), that hardens in 14 min with water as the liquid or 6 min with a 0.25 mol/L sodium phosphate solution as the liquid, without using hydroxyapatite (HA) seeds as setting accelerator, is reported. It was postulated that reduction in porosity would increase cement strength. Thus, the effects of applied pressure during the initial stages of the cement setting reaction on cement strength and porosity were studied. The cement powder comprised an equimolar mixture of TTCP and DCPD (median particle sizes 17 and 1.7 microm, respectively). Compressive strengths (CS) of samples prepared with distilled water were 47.6 +/- 2.4 MPa, 50.7 +/- 4.2 MPa, and 52.9 +/- 4.7 MPa at applied pressures of 5 MPa, 15 MPa, and 25 MPa, respectively. When phosphate solution was used, the CS values obtained were 41.5 +/- 2.3 MPa, 37.9 +/- 1.7 MPa, and 38.1 +/- 2.3 MPa at the same pressure levels. Statistical analysis of the results showed that pressure produced an improvement in CS when water was used as liquid but not when the phosphate solution was used. Compared to previously reported TTCP-DCPD cements, the greater CS values and shorter setting times together with a simplified formulation should make the present TTCP-DCPD cement a useful material as a bone substitute for clinical applications.

  17. Heterogeneous Nucleation of Dicalcium Phosphate Dihydrate on Modified Silica Surfaces.

    PubMed

    Miller, Carrie; Komunjer, Ljepša; Hlady, Vladimir

    2010-01-01

    Heterogeneous nucleation of dicalcium phosphate dihydrate, CaHPO4•2H2O (DCPD) was studied on untreated planar fused silica and on three modified silica surfaces: octadecylsilyl (OTS) modified silica, human serum albumin treated OTS silica, and UV-oxidized 3-mercaptopropyltriethoxysilyl (MTS) modified silica. The supersaturation ratio of calcium and phosphate solution with respect to DCPD was kept below ~10. The nucleated crystals were observed 24 hours and one week after initial contact between supersaturated solutions and substrate surfaces using bright field and reflectance interference contrast microscopy. No DCPD crystals nucleated on albumin-treated OTS-silica. Majority of the DCDP crystals formed on the other modified silica surfaces appeared to be morphologically similar irrespective of the nature of nucleating substrate. Reflectance interference contrast microscopy provided a proof that the majority of the crystals on these substrates do not develop an extended contact with the substrate surface. The images showed that the most extended contact planes were between the DCPD crystals and MTS modified silica surface. The crystals nucleated on OTS-treated and untreated silica surfaces showed only few or none well-developed contact planes.

  18. Inositol pyrophosphates modulate hydrogen peroxide signalling.

    PubMed

    Onnebo, Sara Maria Nancy; Saiardi, Adolfo

    2009-09-14

    Inositol pyrophosphates are involved in a variety of cellular functions, but the specific pathways and/or downstream targets remain poorly characterized. In the present study we use Saccharomyces cerevisiae mutants to examine the potential roles of inositol pyrophosphates in responding to cell damage caused by ROS (reactive oxygen species). Yeast lacking kcs1 [the S. cerevisiae IP6K (inositol hexakisphosphate kinase)] have greatly reduced IP7 (diphosphoinositol pentakisphosphate) and IP8 (bisdiphosphoinositol tetrakisphosphate) levels, and display increased resistance to cell death caused by H2O2, consistent with a sustained activation of DNA repair mechanisms controlled by the Rad53 pathway. Other Rad53-controlled functions, such as actin polymerization, appear unaffected by inositol pyrophosphates. Yeast lacking vip1 [the S. cerevisiae PP-IP5K (also known as IP7K, IP7 kinase)] accumulate large amounts of the inositol pyrophosphate IP7, but have no detectable IP8, indicating that this enzyme represents the physiological IP7 kinase. Similar to kcs1Delta yeast, vip1Delta cells showed an increased resistance to cell death caused by H2O2, indicating that it is probably the double-pyrophosphorylated form of IP8 [(PP)2-IP4] which mediates the H2O2 response. However, these inositol pyrophosphates are not involved in directly sensing DNA damage, as kcs1Delta cells are more responsive to DNA damage caused by phleomycin. We observe in vivo a rapid decrease in cellular inositol pyrophosphate levels following exposure to H2O2, and an inhibitory effect of H2O2 on the enzymatic activity of Kcs1 in vitro. Furthermore, parallel cysteine mutagenesis studies performed on mammalian IP6K1 are suggestive that the ROS signal might be transduced by the direct modification of this evolutionarily conserved class of enzymes.

  19. 21 CFR 182.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Sodium acid pyrophosphate. 182.1087 Section 182.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Food Substances § 182.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b...

  20. 21 CFR 182.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium acid pyrophosphate. 182.1087 Section 182.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Food Substances § 182.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b...

  1. 21 CFR 182.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Sodium acid pyrophosphate. 182.1087 Section 182.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Food Substances § 182.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b...

  2. 21 CFR 582.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Sodium acid pyrophosphate. 582.1087 Section 582.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Additives § 582.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b) Conditions of...

  3. 21 CFR 582.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Sodium acid pyrophosphate. 582.1087 Section 582.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Additives § 582.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b) Conditions of...

  4. 21 CFR 582.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Sodium acid pyrophosphate. 582.1087 Section 582.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Additives § 582.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b) Conditions of...

  5. 21 CFR 582.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Sodium acid pyrophosphate. 582.1087 Section 582.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Additives § 582.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b) Conditions of...

  6. 21 CFR 182.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Sodium acid pyrophosphate. 182.1087 Section 182...) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Multiple Purpose GRAS Food Substances § 182.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate. (b) Conditions of use. This substance is generally...

  7. 21 CFR 582.5306 - Ferric sodium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Ferric sodium pyrophosphate. 582.5306 Section 582.5306 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Dietary Supplements 1 § 582.5306 Ferric sodium pyrophosphate. (a) Product. Ferric sodium pyrophosphate....

  8. 21 CFR 182.6789 - Tetra sodium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Tetra sodium pyrophosphate. 182.6789 Section 182...) SUBSTANCES GENERALLY RECOGNIZED AS SAFE Sequestrants 1 § 182.6789 Tetra sodium pyrophosphate. (a) Product. Tetra sodium pyrophosphate. (b) Conditions of use. This substance is generally recognized as safe...

  9. 21 CFR 582.6789 - Tetra sodium pyrophosphate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Tetra sodium pyrophosphate. 582.6789 Section 582.6789 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED....6789 Tetra sodium pyrophosphate. (a) Product. Tetra sodium pyrophosphate. (b) Conditions of use....

  10. 21 CFR 182.1087 - Sodium acid pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Sodium acid pyrophosphate. 182.1087 Section 182.1087 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... Food Substances § 182.1087 Sodium acid pyrophosphate. (a) Product. Sodium acid pyrophosphate....

  11. Plasma thiamin pyrophosphate and erythrocyte transketolase in chronic alcoholism.

    PubMed

    McLaren, D S; Docherty, M A; Boyd, D H

    1981-06-01

    Thiamin status in patients with an alcohol problem was studied before and after intramuscular thiamin hydrochloride. Results for erythrocyte transketolase activity and plasma thiamin pyrophosphate are compared. Plasma thiamin pyrophosphate values for healthy human subjects are reported for the first time. Advantages of plasma thiamin pyrophosphate in the assessment of thiamin status of patients are discussed.

  12. Pyrophosphate scanning in early frostbite injury

    SciTech Connect

    Purdue, G.F.; Lewis, S.A.; Hunt, J.L.

    1983-11-01

    Early identification of soft tissue injury is a major problem in the patient with frostbite injury. A patient is presented with a method for early (less than 2 days) identification of nonviable tissue. This is a noninvasive method employing technetium 99m stannous pyrophosphate which was used to accurately predict the level of ultimate amputation.

  13. 21 CFR 582.5304 - Ferric pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Ferric pyrophosphate. 582.5304 Section 582.5304 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients and/or...

  14. Effect of the temperature on the synthesis of dicalcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Dhiba, D.; Hakam, A.; El Khoukhi, H.; Albourine, A.; Nounah, A.

    2005-03-01

    The use of calcium phosphates as biomaterials was largely developed during the last decades. Among the phosphorous compounds solid minerals of biological interest, the dicalcium phosphate dihydrate, (DCPD or brushite) (CaHPO{4}, 2H{2}O), disserves a special attention. It can be prepared in different ways. The manufacture of inorganic phosphate, it is usually produced by the acidulation of limestone form of calcium with phosphoric acid. The synthesis of this phosphate depends on many operational parameters. This study was performed in order to verify the influence of temperature and the time on the chemical purity and the yield of precipitation. All the samples were characterized by X-ray diffraction, infrared spectrometry and chemical analysis.

  15. 21 CFR 184.1845 - Stannous chloride (anhydrous and dihydrated).

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... chlorine or gaseous tin tetrachloride. Dihydrated stannous chloride (SnCl2·2H2O, CAS Reg. No. 10025-69-1... granulated tin suspended in water and hydrochloric acid or chlorine. (b) Both forms of the ingredient...

  16. 21 CFR 184.1845 - Stannous chloride (anhydrous and dihydrated).

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... prepared by reacting molten tin with either chlorine or gaseous tin tetrachloride. Dihydrated stannous... molecules of water. It is prepared from granulated tin suspended in water and hydrochloric acid or...

  17. 21 CFR 184.1845 - Stannous chloride (anhydrous and dihydrated).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... prepared by reacting molten tin with either chlorine or gaseous tin tetrachloride. Dihydrated stannous... molecules of water. It is prepared from granulated tin suspended in water and hydrochloric acid or...

  18. 21 CFR 184.1845 - Stannous chloride (anhydrous and dihydrated).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... prepared by reacting molten tin with either chlorine or gaseous tin tetrachloride. Dihydrated stannous... molecules of water. It is prepared from granulated tin suspended in water and hydrochloric acid or...

  19. 21 CFR 184.1845 - Stannous chloride (anhydrous and dihydrated).

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... prepared by reacting molten tin with either chlorine or gaseous tin tetrachloride. Dihydrated stannous... molecules of water. It is prepared from granulated tin suspended in water and hydrochloric acid or...

  20. [Alizarin red S staining of calcium compound crystals in synovial fluid].

    PubMed

    Shoji, K

    1993-04-01

    In order to study the clinical value of alizarin red S staining of calcium compound crystals in synovial fluid, the optimum conditions for staining and the detectable threshold of the crystals were examined. The staining of crystals was mainly affected by the concentration of the dye and pH of the solution. The best results were obtained with an alizarin red S concentration of 1.5-3.0% for hydroxyapatite (HA) and 2.0-3.0% for calcium pyrophosphate dihydrate (CPPD) crystal. The optimal pH of the solution was 4.0-6.0 for HA and 4.0-5.5 for CPPD crystal. For the detection of crystals phagocytosed by synovial fluid leukocytes, staining was enhanced by the addition of chloral hydrate to the dye solution which increased the permeability of the cell membrane. The detectable threshold levels of HA and CPPD crystal in synovial fluid by alizarin red S staining were 0.1 micrograms/ml and 0.5 micrograms/ml, respectively. These results suggest that alizarin red S staining is suitable for screening calcium compound crystals in synovial fluid, because it is more sensitive than other methods, such as polarized microscopy and X-ray diffraction. Alizarin red S staining was performed on 148 synovial fluids from patients with various joint diseases. The staining was positive in 100% of synovial fluids from patients with CPPD deposition disease, in 54% of fluids with osteoarthritis, and in 39% of fluids with rheumatoid arthritis. In osteoarthritis, the increase in the proportion of positive cases was found to be in accordance with the radiological grading of the joints.

  1. Pseudopolymorphism of levodopa: A novel “disappearing” dihydrate

    NASA Astrophysics Data System (ADS)

    André, Vânia; Duarte, M. Teresa

    2014-11-01

    We report herein the crystal structure of a novel L-dopa dihydrate, an unstable pseudopolymorph detected in some co-crystallization studies. This form is obtained by traditional solution techniques and tends to convert to the anhydrous form under ambient conditions. Even though pseudopolymorphism has generally been subject of large conceptual discussions, it is indeed of importance in the pharmaceutical industry and, in this particularly case, the knowledge of the formation of this novel dihydrate may be very relevant for processing issues.

  2. Analysis of Dictyostelium discoideum inositol pyrophosphate metabolism by gel electrophoresis.

    PubMed

    Pisani, Francesca; Livermore, Thomas; Rose, Giuseppina; Chubb, Jonathan Robert; Gaspari, Marco; Saiardi, Adolfo

    2014-01-01

    The social amoeba Dictyostelium discoideum was instrumental in the discovery and early characterization of inositol pyrophosphates, a class of molecules possessing highly-energetic pyrophosphate bonds. Inositol pyrophosphates regulate diverse biological processes and are attracting attention due to their ability to control energy metabolism and insulin signalling. However, inositol pyrophosphate research has been hampered by the lack of simple experimental procedures to study them. The recent development of polyacrylamide gel electrophoresis (PAGE) and simple staining to resolve and detect inositol pyrophosphate species has opened new investigative possibilities. This technology is now commonly applied to study in vitro enzymatic reactions. Here we employ PAGE technology to characterize the D. discoideum inositol pyrophosphate metabolism. Surprisingly, only three major bands are detectable after resolving acidic extract on PAGE. We have demonstrated that these three bands correspond to inositol hexakisphosphate (IP₆ or Phytic acid) and its derivative inositol pyrophosphates, IP₇ and IP₈. Biochemical analyses and genetic evidence were used to establish the genuine inositol phosphate nature of these bands. We also identified IP₉ in D. discoideum cells, a molecule so far detected only from in vitro biochemical reactions. Furthermore, we discovered that this amoeba possesses three different inositol pentakisphosphates (IP₅) isomers, which are largely metabolised to inositol pyrophosphates. Comparison of PAGE with traditional Sax-HPLC revealed an underestimation of the cellular abundance of inositol pyrophosphates by traditional methods. In fact our study revealed much higher levels of inositol pyrophosphates in D. discoideum in the vegetative state than previously detected. A three-fold increase in IP₈ was observed during development of D. discoideum a value lower that previously reported. Analysis of inositol pyrophosphate metabolism using ip6k null amoeba

  3. Analysis of Dictyostelium discoideum Inositol Pyrophosphate Metabolism by Gel Electrophoresis

    PubMed Central

    Pisani, Francesca; Livermore, Thomas; Rose, Giuseppina; Chubb, Jonathan Robert; Gaspari, Marco; Saiardi, Adolfo

    2014-01-01

    The social amoeba Dictyostelium discoideum was instrumental in the discovery and early characterization of inositol pyrophosphates, a class of molecules possessing highly-energetic pyrophosphate bonds. Inositol pyrophosphates regulate diverse biological processes and are attracting attention due to their ability to control energy metabolism and insulin signalling. However, inositol pyrophosphate research has been hampered by the lack of simple experimental procedures to study them. The recent development of polyacrylamide gel electrophoresis (PAGE) and simple staining to resolve and detect inositol pyrophosphate species has opened new investigative possibilities. This technology is now commonly applied to study in vitro enzymatic reactions. Here we employ PAGE technology to characterize the D. discoideum inositol pyrophosphate metabolism. Surprisingly, only three major bands are detectable after resolving acidic extract on PAGE. We have demonstrated that these three bands correspond to inositol hexakisphosphate (IP6 or Phytic acid) and its derivative inositol pyrophosphates, IP7 and IP8. Biochemical analyses and genetic evidence were used to establish the genuine inositol phosphate nature of these bands. We also identified IP9 in D. discoideum cells, a molecule so far detected only from in vitro biochemical reactions. Furthermore, we discovered that this amoeba possesses three different inositol pentakisphosphates (IP5) isomers, which are largely metabolised to inositol pyrophosphates. Comparison of PAGE with traditional Sax-HPLC revealed an underestimation of the cellular abundance of inositol pyrophosphates by traditional methods. In fact our study revealed much higher levels of inositol pyrophosphates in D. discoideum in the vegetative state than previously detected. A three-fold increase in IP8 was observed during development of D. discoideum a value lower that previously reported. Analysis of inositol pyrophosphate metabolism using ip6k null amoeba revealed

  4. Solubility of dicalcium phosphate dihydrate by solid titration.

    PubMed

    Pan, H-B; Darvell, B W

    2009-01-01

    Solid-titration results for hydroxyapatite (HAp), octacalcium phosphate, beta-tricalcium phosphate and tetracalcium phosphate have shown that the only stable phase in 100 mmol x l(-1) KCl at 37 degrees C is HAp. In particular, dicalcium phosphate dihydrate (DCPD) did not form at pH <4.2 (where it is otherwise believed to be stable) except as a metastable phase under conditions of slight supersaturation. The behaviour of DCPD itself under the same conditions requires checking. Solid titration was used to determine the apparent solubility of DCPD in a 100-mmol x l(-1) KCl solution at 37.0 +/- 0.1 degrees C over the pH range 3.2-11.6. The constitution of the precipitate was determined by X-ray diffraction, particle morphology was observed by scanning and transmission electron microscopy, and the precipitate Ca/P ratio was calculated by energy-dispersive X-ray analysis. The titration curve for DCPD was substantially lower than the position reported elsewhere. DCPD was the only identified phase at equilibrium at pH 3.60 and 4.50, but HAp was formed after seeding with 1 mg HAp at DCPD equilibrium at pH 4.47, 3.60 and 3.30. It is concluded that the titration curve observed for DCPD corresponds to the solubility isotherm for the phase, but that this represents a metastable equilibrium. HAp is more stable than DCPD, particularly below pH 4.2. The implications for calcium phosphate studies are profound as the reverse is generally believed to be true. Thus, solubility results and the nature of the carious lesion need reconsideration.

  5. In vitro formation of dicalcium phosphate dihydrate, CaHPO4 . 2H2O (DCPD).

    PubMed

    LeGeros, R Z; Lee, D; Quirolgico, G; Shirra, W P; Reich, L

    1983-01-01

    Dicalcium phosphate dihydrate, CaHPO4 . 2H2O (DCPD) crystals were grown in solutions and in different types of gels (silica, agar, collagen) at different pH's, in the presence of other ions, and on different Ca-bearing substrates (enamel, dentine, bone, calcite). Hydrolysis of DCPD to other calcium phosphates and calcium oxalates were made in solutions containing different ions at 37 degrees C, 24h. Results showed that the type of media (solution or gel), type of gel, pH, presence of other ions affect the morphology (size/shape) and growth of DCPD. In systems of changing pH, the more basic calcium phosphate (e.g., apatite, OCP) formed in the first zone, DCPD in the last zone, regardless of the initial Ca/P of the system. Growth of DCPD was suppressed in the presence of F-(which favored growth of apatite), of Cd2+, Pb2+ (which favored OCP). Hydrolysis of DCPD to apatite was promoted in solutions containing Ca2+, F-, CO3=, and inhibited by Mg2+, P2O74-. Hydrolysis of DCPD to calcium oxalate occurs in presence of C2O4=. Results indicate that the variation in morphology of DCPD occurring in various human tissue calcinosis and the co-existence of DCPD with other calcium phosphates and oxalates in urinary and dental calculi are due to variations in pH, and presence of different ions in the media.

  6. Synthesis and biological evaluation of pyrophosphate mimics of thiamine pyrophosphate based on a triazole scaffold.

    PubMed

    Erixon, Karl M; Dabalos, Chester L; Leeper, Finian J

    2008-10-07

    Novel triazole-based pyrophosphate analogues of thiamine pyrophosphate (TPP) have been synthesised and tested for inhibition of pyruvate decarboxylase (PDC) from Zymomonas mobilis. The thiazolium ring of thiamine was replaced by a triazole in an efficient two-step procedure. Pyrophosphorylation then gave extremely potent triazole inhibitors with K(I) values down to 20 pM, compared to a K(D) value of 0.35 microM for TPP. This triazole scaffold was used for further investigation and six analogues containing mimics of the pyrophosphate group were synthesised and tested for inhibition of PDC. Several effective analogues were found with K(I) values down to around 1 nM.

  7. Hydrogen bonding. Part 82. Thermodynamic and infrared study of dimethonium and pentamethonium halide dihydrates

    NASA Astrophysics Data System (ADS)

    Harmon, Kenneth M.; Nikolla, Eranda

    2003-09-01

    We have carried out equilibrium dissociation vapor pressure measurements on dimethonium bromide dihydrate and pentamethonium chloride and bromide dihydrates. None of these salts forms a monohydrate. Surprisingly the thermodynamic parameters for the two pentamethonium hydrates are nearly identical, although the properties of these hydrates are quite different. This is explained by a larger negative differential lattice enthalpy for the chloride dihydrate dissociation, which lowers the observed enthalpy of H 2O removal, and greater stabilization of saturated solution by chloride ion which makes the chloride dihydrate deliquescent while the bromide dihydrate is efflorescent. Infrared comparison suggests that tetramethonium chloride dihydrate and pentamethonium chloride and bromide dihydrates have the ladder type halide-water structure determined by X-ray analysis in tetramethonium bromide dihydrate.

  8. Substitution of calcium by strontium within selected calcium phosphates

    NASA Astrophysics Data System (ADS)

    Rokita, E.; Hermes, C.; Nolting, H.-F.; Ryczek, J.

    1993-06-01

    Sr incorporation in the molecules of amorphous calcium phosphate, apatitic tricalcium phosphate, hydroxyapatite, octacalcium phosphate and dicalcium phosphate dihydrate was investigated. The concentration of Sr ranged from 225 to 1010 μ g / g, i.e. it overlapped with the physiological range of Sr concentrations in human bone. The leading experimental technique was extended X-ray absorption fine structure (EXAFS) at the Sr K edge. Results of these studies demonstrated the following: (1) Sr incorporation in the calcium phosphates is compound-dependent, (2) the coordination of incorporated Sr atoms in the Ca-P molecules is similar to that of Ca atoms, but interatomic distances are ≈0.015 nm larger, (3) in apatitic tricalcium phosphate, hydroxyapatite and octacalcium phosphate lattices Sr atoms may occupy selected Ca sites, which was not the case for dicalcium phosphate dihydrate, (4) in the apatite lattice Sr atoms are coordinated by 6 PO 4 tetrahedrals and (5) EXAFS spectra at the K edge of the incorporated Sr may be used to distinguish the structures of amorphous calcium phosphate, dicalcium phosphate dihydrate as well as apatite and its derivatives (apatitic tricalcium phosphate, octacalcium phosphate).

  9. Formation of ettringite, Ca 6Al 2(SO 4) 3(OH) 12·26H 2O, AFt, and monosulfate, Ca 4Al 2O 6(SO 4)·14H 2O, AFm-14, in hydrothermal hydration of Portland cement and of calcium aluminum oxide—calcium sulfate dihydrate mixtures studied by in situ synchrotron X-ray powder diffraction

    NASA Astrophysics Data System (ADS)

    Christensen, Axel Nørlund; Jensen, Torben R.; Hanson, Jonathan C.

    2004-06-01

    In the hydration of calcium aluminum oxide-gypsum mixtures, i.e., Ca 3Al 2O 6, Ca 12Al 14O 33 and CaSO 4·2H 2O, the reaction products can be ettringite, Ca 6Al 2(SO 4) 3(OH) 12·26H 2O, monosulfate, Ca 4Al 2O 6(SO 4)·14H 2O, or the calcium aluminum oxide hydrate, Ca 4Al 2O 7·19H 2O. Ettringite is formed if sufficient CaSO 4·2H 2O is present in the mixture. Ettringite is converted to monosulfate when all CaSO 4·2H 2O is consumed in the synthesis of ettringite. The reactions were investigated in the temperature range 25-170°C using in situ synchrotron X-ray powder diffraction. This technique allows the study of very fast chemical reactions that are observed here under hydrothermal conditions. A new experimental approach was developed to perform in situ mixing of the reactants during X-ray data collection.

  10. Enrofloxacin hydro­chloride dihydrate

    PubMed Central

    Miranda-Calderón, Jorge E.; Gutiérrez, Lilia; Flores-Alamo, Marcos; García-Gutiérrez, Ponciano; Sumano, Héctor

    2014-01-01

    The asymmetric unit of the title compound, C19H23FN3O3 +·Cl−·2H2O [systematic name: 4-(3-carb­oxy-1-cyclo­propyl-6-fluoro-4-oxo-1,4-di­hydro­quin­o­lin-7-yl)-1-ethyl­piperazin-1-ium chloride dihydrate], consists of two independent monocations of the protonated enrofloxacin, two chloride anions and four water mol­ecules. In the cations, the piperazinium rings adopt chair conformations and the dihedral angles between the cyclo­propyl ring and the 10-membered quinoline ring system are 56.55 (2) and 51.11 (2)°. An intra­molecular O—H⋯O hydrogen bond is observed in each cation. In the crystal, the components are connected via O—H⋯Cl, N—H⋯Cl and O—H⋯O hydrogen bonds, and a π–π inter­action between the benzene rings [centroid–centroid distance = 3.6726 (13) Å], resulting in a three-dimensional array. PMID:24826167

  11. A comparison of trehalose dihydrate and mannitol as stabilizing agents for dicalcium phosphate dihydrate based tablets.

    PubMed

    Landín, M; Fontao, M J; Martínez-Pacheco, R

    2005-03-01

    This study investigated the possible utility of trehalose dihydrate (TD) as a tablet stabilizing agent. Acetylsalicylic acid was used as the model hydrolyzable drug and dicalcium phosphate dihydrate (DCPD) as the base excipient, because it is well documented that ASA/DCPD tablets are unstable during storage at low temperature and high relative humidity; DCPD is usually combined with mannitol in order to improve tablet stability. Tablets comprising DCPD, 10% ASA, and 0%, 10%, or 20% w/w of TD were prepared by direct compression and stored at 35 degrees C and 82.9% relative humidity for 6 months. Additionally, control tablets with DCPD and ASA, only, or with DCPD, ASA and 20% mannitol, were also evaluated. At predetermined time intervals, formulations were tested for drug content, mechanical, microstructural, and drug dissolution properties. Additionally, thermal analyses and ASA solution stability studies were carried out. Results reveal that both TD and mannitol significantly reduce degradation of ASA included in DCPD-based tablets, but neither effectively protects against the marked decline in tablet mechanical properties on aging. The ASA stabilization effects of TD and mannitol were also observed in solution, indicating an interaction between these sugars and ASA.

  12. Effects of ethanol addition on formation of hydroxyapatite through hydrothermal treatment of dicalcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Goto, T.; Kamitakahara, M.; Kim, I. Y.; Ohtsuki, C.

    2011-10-01

    The mixture of dicalcium phosphate dihydrate (DCPD) and calcium acetate monohydrate were hydrothermally treated in a condition of water-ethanol mixed solvent at 120 °C for various periods. The rate of hydroxyapatite (HAp) formation was decreased with increasing the volume ratio of ethanol, to result in formation of dicalcium phosphate anhydrous and β-tricalcium phosphate. Needle-like HAp particles were observed in the sample treated with the mere water. The sample treated with the ethanol-water mixed solvent had nano-sized HAp particles with a form of the plate-like crystals. The size of HAp crystal was decreased with increasing the fraction of ethanol. These results show that HAp formation and crystal growth were prevented by the ethanol addition.

  13. Inositol pyrophosphates inhibit synaptotagmin-dependent exocytosis

    PubMed Central

    Lee, Tae-Sun; Lee, Joo-Young; Kyung, Jae Won; Yang, Yoosoo; Park, Seung Ju; Lee, Seulgi; Pavlovic, Igor; Kong, Byoungjae; Jho, Yong Seok; Jessen, Henning J.; Kweon, Dae-Hyuk; Shin, Yeon-Kyun; Kim, Sung Hyun; Yoon, Tae-Young; Kim, Seyun

    2016-01-01

    Inositol pyrophosphates such as 5-diphosphoinositol pentakisphosphate (5-IP7) are highly energetic inositol metabolites containing phosphoanhydride bonds. Although inositol pyrophosphates are known to regulate various biological events, including growth, survival, and metabolism, the molecular sites of 5-IP7 action in vesicle trafficking have remained largely elusive. We report here that elevated 5-IP7 levels, caused by overexpression of inositol hexakisphosphate (IP6) kinase 1 (IP6K1), suppressed depolarization-induced neurotransmitter release from PC12 cells. Conversely, IP6K1 depletion decreased intracellular 5-IP7 concentrations, leading to increased neurotransmitter release. Consistently, knockdown of IP6K1 in cultured hippocampal neurons augmented action potential-driven synaptic vesicle exocytosis at synapses. Using a FRET-based in vitro vesicle fusion assay, we found that 5-IP7, but not 1-IP7, exhibited significantly higher inhibitory activity toward synaptic vesicle exocytosis than IP6. Synaptotagmin 1 (Syt1), a Ca2+ sensor essential for synaptic membrane fusion, was identified as a molecular target of 5-IP7. Notably, 5-IP7 showed a 45-fold higher binding affinity for Syt1 compared with IP6. In addition, 5-IP7–dependent inhibition of synaptic vesicle fusion was abolished by increasing Ca2+ levels. Thus, 5-IP7 appears to act through Syt1 binding to interfere with the fusogenic activity of Ca2+. These findings reveal a role of 5-IP7 as a potent inhibitor of Syt1 in controlling the synaptic exocytotic pathway and expand our understanding of the signaling mechanisms of inositol pyrophosphates. PMID:27364007

  14. Thermodynamic examination of the pyrophosphate sensor helix in the thiamine pyrophosphate riboswitch.

    PubMed

    Furniss, Stephanie; Grover, Neena

    2011-04-01

    Riboswitches are functional mRNA that control gene expression. Thiamine pyrophosphate (TPP) binds to thi-box riboswitch RNA and allosterically inhibits genes that code for proteins involved in the biosynthesis and transport of thiamine. Thiamine binding to the pyrimidine sensor helix and pyrophosphate binding to the pyrophosphate sensor helix cause changes in RNA conformation that regulate gene expression. Here we examine the thermodynamic properties of the internal loop of the pyrophosphate binding domain by comparing the wild-type construct (RNA WT) with six modified 2 x 2 bulged RNA and one 2 x 2 bulged DNA. The wild-type construct retains five conserved bases of the pyrophosphate sensor domain, two of which are in the 2 x 2 bulge (C65 and G66). The RNA WT construct was among the most stable (ΔG°₃₇ = -7.7 kcal/mol) in 1 M KCl at pH 7.5. Breaking the A•G mismatch of the bulge decreases the stability of the construct ~0.5-1 kcal/mol, but does not affect magnesium binding to the RNA WT. Guanine at position 48 is important for RNA-Mg²+ interactions of the TPP-binding riboswitch at pH 7.5. In the presence of 9.5 mM magnesium at pH 5.5, the bulged RNA constructs gained an average of 1.1 kcal/mol relative to 1 M salt. Formation of a single A+•C mismatch base pair contributes about 0.5 kcal/mol at pH 5.5, whereas two tandem A+•C mismatch base pairs together contribute about 2 kcal/mol.

  15. Two inositol hexakisphosphate kinases drive inositol pyrophosphate synthesis in plants

    USDA-ARS?s Scientific Manuscript database

    Inositol pyrophosphates are novel cellular signaling molecules with newly discovered roles in energy sensing and metabolic control. Studies in eukaryotes have revealed that these compounds turn over rapidly, and thus only small amounts accumulate. Inositol pyrophosphates have not been the subject of...

  16. Formation of pyrophosphate on hydroxyapatite with thioesters as condensing agents

    NASA Technical Reports Server (NTRS)

    Weber, A. L.

    1982-01-01

    'Energy-rich' thioesters are shown to act as condensing agents in the formation of pyrophosphate on hydroxyapatite in the presence of water at ambient temperature. The yield of pyrophosphate based on thioester ranges from 2.5% to 11.4% and depends upon the pH and concentration of reactants. Reaction of 0.130 M hydroxyapatite suspended in a solution of 0.08 M sodium phosphate and 0.20 M imidazole hydrochloride (pH 7.0) with 0.10 M N,S-diacetylcysteamine for 6 days gives the highest yield of pyrophosphate (11.4%). Pyrophosphate formation requires the presence of hydroxyapatite, sodium phosphate and the thioester, N,S-diacetylcysteamine. The related thioester, N,S-diacetylcysteine, also yields pyrophosphate in reactions on hydroxyapatite.

  17. Effect of tetrasodium pyrophosphate concentration and cooking time on the physicochemical properties of process cheese.

    PubMed

    Shirashoji, N; Aoyagi, H; Jaeggi, J J; Lucey, J A

    2016-09-01

    Tetrasodium pyrophosphate (TSPP) is widely used as an emulsifying salt (ES) in process cheese. Previous reports have indicated that TSPP exhibits some unusual properties, including the gelation of milk proteins at specific ES concentrations. We studied the effect of various concentrations (0.25-2.75%) of TSPP and cooking times (0-20min) on the rheological, textural, and physical properties of pasteurized process Cheddar cheese using a central composite rotatable experimental design. Cheeses were made with a constant pH value to avoid pH as a confounding factor. Modeling of the textural properties of process cheese made with TSPP exhibited complex behavior, with polynomial models (cubic) giving better predictions (higher coefficient of determination values) than simpler quadratic models. Meltability indices (degree of flow from the UW MeltProfiler (University of Wisconsin-Madison), loss tangent value at 60°C from rheological testing, and Schreiber melt area) initially decreased with increasing TSPP concentrations, but above a critical ES concentration (~1.0%) meltability increased at higher TSPP concentrations. The storage modulus values measured at 70°C for process cheese initially increased with increasing TSPP concentration, but above a concentration of 1% ES, the storage modulus values decreased. Cooking time had little effect on the various melting or rheological properties. With an increase in TSPP concentration, the insoluble Ca and P contents increased, suggesting that TSPP addition resulted in the formation of insoluble calcium pyrophosphate complexes; some of which were likely associated with caseins. A portion of the added TSPP remained in the soluble phase. The acid-base buffering profiles also indicated that calcium pyrophosphate complexes were formed in cheese made with TSPP. In milk systems, low levels of TSPP have been shown to induce protein crosslinking and gelation, whereas at higher TSPP concentrations milk gelation was inhibited due to

  18. Identification of dicalcium phosphate dihydrate deposited during osteoblast mineralization in vitro.

    PubMed

    Zhang, Zheng-Lai; Chen, Xiao-Rong; Bian, Sha; Huang, Jian; Zhang, Tian-Lan; Wang, Kui

    2014-02-01

    The hydroxyapatite (HAP) with variable chemical substitutions has been considered as the major component in the mineralized part of bones. Various metastable crystalline phases have been suggested as transitory precursors of HAP in bone, but there are no consensuses as to the nature of these phases and their temporal evolution. In the present study, we cultured rat calvarial osteoblasts with ascorbate and β-glycerophosphate to explore which calcium phosphate precursor phases comprise the initial mineral in the process of osteoblast mineralization in vitro. At the indicated time points, the deposited calcium phosphate was analyzed after removing organic substances from the extracellular matrix with hydrazine. The features comparable to dicalcium phosphate dihydrate (DCPD) and octacalcium phosphate (OCP), in addition to HAP, were detected in the mineral phases by high resolution transmission electron microscopy. And there was a trend of conversion from DCPD- and OCP-like phases to HAP in the course of mineralization, as indicated by Fourier-transform infrared microspectroscopy, energy-dispersive X-ray spectroscopy and synchrotron X-ray powder diffraction analyses. Besides, biochemical assay showed a progressive decrease in the ratio of mineral-associated proteins to calcium with time. These findings suggest that DCPD- and OCP-like phases are likely to occur on the course of osteoblast mineralization, and the mineral-associated proteins might be involved in modulating the mineral phase transformation.

  19. Growth, spectral and thermal properties of manganous malonate dihydrate crystals

    NASA Astrophysics Data System (ADS)

    Lincy, A.; Mahalakshmi, V.; Thomas, J.; Saban, K. V.

    2012-02-01

    Manganous malonate dihydrate crystals have been grown by ionic diffusion in hydrosilica gel. Powder XRD pattern and the FTIR spectrum have been recorded for the sample. The vibrational bands corresponding to various functional groups are identified. Thermogravimetric studies have been done to explore the nature of the material as regards thermal decomposition.

  20. Uranium pyrophosphate / methylenediphosphonate polyoxometalate cage clusters

    SciTech Connect

    Ling, Jie; Qiu, Jie; Sigmon, Ginger E.; Ward, Matt; Szymanowski, Jennifer E.S.; Burns, Peter C

    2010-09-29

    Despite potential applications in advanced nuclear energy systems, nanoscale control of uranium materials is in its infancy. In its hexavalent state, U occurs as (UO{sub 2}){sup 2+} uranyl ions that are coordinated by various ligands to give square, pentagonal, or hexagonal bipyramids. Creation and design of nanostructured uranyl materials requires interruption of the tendency of uranyl bipyramids to share equatorial edges to form infinite sheets that occur in extended structures. Where a bidentate peroxide group bridges uranyl bipyramids, the configuration is inherently bent, fostering formation of cage clusters. Here the bent configurations of four- and five-membered rings of uranyl peroxide hexagonal bipyramids are bridged by pyrophosphate or methylenediphosphonate, creating eight chemically complex cage clusters with specific topologies. Chemical complexity in such clusters provides opportunities for the tuning of cage sizes, pore sizes, and properties such as aqueous solubility. Several of these are topological derivatives of simpler clusters that contain only uranyl bipyramids, whereas others exhibit new topologies.

  1. The purification of 3,3-dimethylallyl- and geranyl-transferase and of isopentenyl pyrophosphate isomerase from pig liver

    PubMed Central

    Holloway, P. W.; Popják, G.

    1967-01-01

    The enzyme catalysing the synthesis of farnesyl pyrophosphate from dimethylallyl pyrophosphate and isopentenyl pyrophosphate, or from geranyl pyrophosphate and isopentenyl pyrophosphate, has been purified 100-fold from homogenates of pig liver. The enzyme has optimum pH 7·9 and requires Mg2+ as activator in preference to Mn2+; it is inhibited by iodoacetamide, N-ethylmaleimide, p-hydroxymercuribenzoate and phosphate ions in addition to the products of the reaction, inorganic pyrophosphate and farnesyl pyrophosphate. From product-inhibition studies of the geranyltransferase reaction, the order of addition of substrates to and release of products from the enzyme has been deduced: geranyl pyrophosphate combines with the enzyme first, followed by isopentenyl pyrophosphate. Farnesyl pyrophosphate dissociates from the enzyme before inorganic pyrophosphate. The existence of isopentenyl pyrophosphate isomerase in liver is confirmed. Methods for the preparation of the pyrophosphate esters of isopentenol, 3,3-dimethylallyl alcohol, geraniol and farnesol are also described. PMID:4292002

  2. Estimation of crystallinity of trehalose dihydrate microspheres by usage of terahertz time-domain spectroscopy.

    PubMed

    Takeuchi, Issei; Tomoda, Keishiro; Nakajima, Takehisa; Terada, Hiroshi; Kuroda, Hideki; Makino, Kimiko

    2012-09-01

    Crystalline state of pharmaceutical materials is of great importance in the preparation of pharmaceutics because their physicochemical properties affect bioavailability, quality of products, therapeutic level, and manufacturing process. In this study, we have estimated the crystallinity of trehalose dihydrate microspheres by measuring terahertz (THz) spectroscopy. The commercially available trehalose dihydrate takes in general a crystalline state, but trehalose dihydrate microspheres prepared by using spray-drying method are in an amorphous state. We have prepared amorphous anhydrous trehalose by using melt-quenched method from crystalline trehalose dihydrate. We have measured the absorbance of trehalose dihydrate containing amorphous anhydrous trehalose (0%, 25%, 50%, 75%, and 100%) using THz time-domain spectroscopy (THz-TDS) to prepare calibration curves. Using the calibration curves, we have estimated the crystallinity of trehalose dihydrate microspheres prepared by using spray-drying method. Our results suggest that THz-TDS is well suited to distinguish crystallinity differences in pharmaceutical compounds.

  3. A Novel Inositol Pyrophosphate Phosphatase in Saccharomyces cerevisiae

    PubMed Central

    Steidle, Elizabeth A.; Chong, Lucy S.; Wu, Mingxuan; Crooke, Elliott; Fiedler, Dorothea; Resnick, Adam C.; Rolfes, Ronda J.

    2016-01-01

    Inositol pyrophosphates are high energy signaling molecules involved in cellular processes, such as energetic metabolism, telomere maintenance, stress responses, and vesicle trafficking, and can mediate protein phosphorylation. Although the inositol kinases underlying inositol pyrophosphate biosynthesis are well characterized, the phosphatases that selectively regulate their cellular pools are not fully described. The diphosphoinositol phosphate phosphohydrolase enzymes of the Nudix protein family have been demonstrated to dephosphorylate inositol pyrophosphates; however, the Saccharomyces cerevisiae homolog Ddp1 prefers inorganic polyphosphate over inositol pyrophosphates. We identified a novel phosphatase of the recently discovered atypical dual specificity phosphatase family as a physiological inositol pyrophosphate phosphatase. Purified recombinant Siw14 hydrolyzes the β-phosphate from 5-diphosphoinositol pentakisphosphate (5PP-IP5 or IP7) in vitro. In vivo, siw14Δ yeast mutants possess increased IP7 levels, whereas heterologous SIW14 overexpression eliminates IP7 from cells. IP7 levels increased proportionately when siw14Δ was combined with ddp1Δ or vip1Δ, indicating independent activity by the enzymes encoded by these genes. We conclude that Siw14 is a physiological phosphatase that modulates inositol pyrophosphate metabolism by dephosphorylating the IP7 isoform 5PP-IP5 to IP6. PMID:26828065

  4. Thermal and dielectric studies of nickel malonate dihydrate single crystals

    NASA Astrophysics Data System (ADS)

    Mathew, Varghese; Mathai, K. C.; Mahadeven, C. K.; Abraham, K. E.

    2011-02-01

    Single crystals of nickel malonate dihydrate were grown by the gel technique, employing the single diffusion method. Thermal dehydration of the crystal was investigated by thermogravimetric and differential thermal analyses. The title compound exhibits a steady thermal behaviour at higher temperature range of 350-800 °C. The dielectric properties of the prepared sample were analyzed as a function of frequency in the range of 1 kHz-1 MHz and at temperatures between 40 and 140 °C.

  5. Inositol pyrophosphates: why so many phosphates?

    PubMed Central

    Shears, Stephen B.

    2014-01-01

    The inositol pyrophosphates (PP-InsPs) are a specialized group of “energetic” signaling molecules found in yeasts, plants and animals. PP-InsPs boast the most crowded three dimensional phosphate arrays found in Nature; multiple phosphates and diphosphates are crammed around the six-carbon, inositol ring. Yet, phosphate esters are also a major energy currency in cells. So the synthesis of PP-InsPs, and the maintenance of their levels in the face of a high rate of ongoing turnover, all requires significant bioenergetic input. What are the particular properties of PP-InsPs that repay this investment of cellular energy? Potential answers to that question are discussed here, against the backdrop of a recent hypothesis that signaling by PP-InsPs is evolutionarily ancient. The latter idea is extended herein, with the proposal that the primordial origins of PP-InsPs is reflected in the apparent lack of isomeric specificity of certain of their actions. Nevertheless, there are other aspects of signaling by these polyphosphates that are more selective for a particular PP-InsP isomer. Consideration of the nature of both specific and non-specific effects of PP-InsPs can help rationalize why such molecules possess so many phosphates. PMID:25453220

  6. PLUTONIUM PURIFICATION PROCESS EMPLOYING THORIUM PYROPHOSPHATE CARRIER

    DOEpatents

    King, E.L.

    1959-04-28

    The separation and purification of plutonium from the radioactive elements of lower atomic weight is described. The process of this invention comprises forming a 0.5 to 2 M aqueous acidffc solution containing plutonium fons in the tetravalent state and elements with which it is normally contaminated in neutron irradiated uranium, treating the solution with a double thorium compound and a soluble pyrophosphate compound (Na/sub 4/P/sub 2/O/sub 7/) whereby a carrier precipitate of thorium A method is presented of reducing neptunium and - trite is advantageous since it destroys any hydrazine f so that they can be removed from solutions in which they are contained is described. In the carrier precipitation process for the separation of plutonium from uranium and fission products including zirconium and columbium, the precipitated blsmuth phosphate carries some zirconium, columbium, and uranium impurities. According to the invention such impurities can be complexed and removed by dissolving the contaminated carrier precipitate in 10M nitric acid, followed by addition of fluosilicic acid to about 1M, diluting the solution to about 1M in nitric acid, and then adding phosphoric acid to re-precipitate bismuth phosphate carrying plutonium.

  7. Uropontin in urinary calcium stone formation.

    PubMed

    Hoyer, J R

    1994-01-01

    Normal urine is frequently supersaturated with respect to calcium oxalate. Thus, urinary inhibitors of crystallization appear to have an important role in preventing urinary stone formation. Uropontin was isolated by monoclonal antibody immunoaffinity chromatography and has the same N-terminal sequence as osteopontin derived from bone. This urinary form of osteopontin is a potent inhibitor of calcium oxalate monohydrate crystal growth at concentrations (approximately 0.1 microM) that normally prevail in human urine. Interaction with calcium oxalate monohydrate in vivo was shown by analysis of EDTA extracts of calcium stones. Uropontin is an abundant component of calcium oxalate monohydrate stones and present in only trace quantities in calcium oxalate dihydrate and hydroxyapatite stones. However, the precise role of uropontin in the pathogenesis of urinary stone formation is not known and is the subject of ongoing investigations.

  8. Proposed Carrier Lipid-binding Site of Undecaprenyl Pyrophosphate Phosphatase from Escherichia coli*

    PubMed Central

    Chang, Hsin-Yang; Chou, Chia-Cheng; Hsu, Min-Feng; Wang, Andrew H. J.

    2014-01-01

    Undecaprenyl pyrophosphate phosphatase (UppP), an integral membrane protein, catalyzes the dephosphorylation of undecaprenyl pyrophosphate to undecaprenyl phosphate, which is an essential carrier lipid in the bacterial cell wall synthesis. Sequence alignment reveals two consensus regions, containing glutamate-rich (E/Q)XXXE plus PGXSRSXXT motifs and a histidine residue, specific to the bacterial UppP enzymes. The predicted topological model suggests that both of these regions are localized near the aqueous interface of UppP and face the periplasm, implicating that its enzymatic function is on the outer side of the plasma membrane. The mutagenesis analysis demonstrates that most of the mutations (E17A/E21A, H30A, S173A, R174A, and T178A) within the consensus regions are completely inactive, indicating that the catalytic site of UppP is constituted by these two regions. Enzymatic analysis also shows an absolute requirement of magnesium or calcium ions in enzyme activity. The three-dimensional structural model and molecular dynamics simulation studies have shown a plausible structure of the catalytic site of UppP and thus provides insights into the molecular basis of the enzyme-substrate interaction in membrane bilayers. PMID:24855653

  9. Red blood cell membrane fragments but not intact red blood cells promote calcium oxalate monohydrate crystal growth and aggregation.

    PubMed

    Chutipongtanate, Somchai; Thongboonkerd, Visith

    2010-08-01

    Cell membranes are thought to promote calcium oxalate kidney stone formation but to our knowledge the modulating effect of red blood cell membranes on calcium oxalate crystals has not been previously investigated. Thus, we examined the effects of red blood cell membrane fragments on calcium oxalate monohydrate and calcium oxalate dihydrate crystal growth and aggregation. Calcium oxalate monohydrate and calcium oxalate dihydrate crystals were treated with red blood cell membrane fragments or intact red blood cells from a healthy donor. Phase contrast microscopy was performed to evaluate crystal morphology and aggregation. We used ImageMaster 2D Platinum software to evaluate crystal size and spectrophotometric oxalate depletion assay to monitor crystal growth. Red blood cell membrane fragments had significant promoting activity on calcium oxalate monohydrate crystal growth with an approximately 75% increase in size and aggregation with an approximately 2.5-fold increase in aggregate number compared to the control without membrane fragments or cells. Approximately 50% of calcium oxalate monohydrate crystals were adhered by red blood cell membrane fragments. Intact red blood cells had no significant effect on calcium oxalate monohydrate crystal growth or aggregation but they could transform calcium oxalate monohydrate to calcium oxalate dihydrate crystals. Red blood cell membrane fragments and intact red blood cells had no effect on calcium oxalate dihydrate crystals. The promoting activity of red blood cell membrane fragments on calcium oxalate monohydrate crystal growth was successfully confirmed by spectrophotometric oxalate depletion assay. To our knowledge our data provide the first direct evidence that red blood cell membrane fragments are a promoting factor for calcium oxalate monohydrate crystal growth and aggregation. Thus, they may aggravate calcium oxalate stone formation. Copyright (c) 2010 American Urological Association Education and Research, Inc

  10. Revisiting spatial distribution and biochemical composition of calcium-containing crystals in human osteoarthritic articular cartilage

    PubMed Central

    2013-01-01

    Introduction Calcium-containing (CaC) crystals, including basic calcium phosphate (BCP) and calcium pyrophosphate dihydrate (CPP), are associated with destructive forms of osteoarthritis (OA). We assessed their distribution and biochemical and morphologic features in human knee OA cartilage. Methods We prospectively included 20 patients who underwent total knee replacement (TKR) for primary OA. CaC crystal characterization and identification involved Fourier-transform infra-red spectrometry and scanning electron microscopy of 8 to 10 cartilage zones of each knee, including medial and lateral femoral condyles and tibial plateaux and the intercondyle zone. Differential expression of genes involved in the mineralization process between cartilage with and without calcification was assessed in samples from 8 different patients by RT-PCR. Immunohistochemistry and histology studies were performed in 6 different patients. Results Mean (SEM) age and body mass index of patients at the time of TKR was 74.6 (1.7) years and 28.1 (1.6) kg/m², respectively. Preoperative X-rays showed joint calcifications (chondrocalcinosis) in 4 cases only. The medial femoro-tibial compartment was the most severely affected in all cases, and mean (SEM) Kellgren-Lawrence score was 3.8 (0.1). All 20 OA cartilages showed CaC crystals. The mineral content represented 7.7% (8.1%) of the cartilage weight. All patients showed BCP crystals, which were associated with CPP crystals for 8 joints. CaC crystals were present in all knee joint compartments and in a mean of 4.6 (1.7) of the 8 studied areas. Crystal content was similar between superficial and deep layers and between medial and femoral compartments. BCP samples showed spherical structures, typical of biological apatite, and CPP samples showed rod-shaped or cubic structures. The expression of several genes involved in mineralization, including human homolog of progressive ankylosis, plasma-cell-membrane glycoprotein 1 and tissue

  11. Synthesis and conductivity of indium-doped tin pyrophosphates

    SciTech Connect

    Garzon, Fernando H; Mukundan, Rangachary; Brosha, Eric L

    2008-01-01

    We have synthesized indium-doped tin pyrophosphates as high-temperature anhydrous proton conductors. The ratio of tin to indium was varied using two different synthetic methods. The first is a high-temperature reaction in which a paste containing the reactants in excess phosphoric acid was heated for various amounts of time at various temperatures. The second method is a solution precipitation procedure followed by calcination, which offers several advantages over traditional synthetic techniques. These advantages inc 1 ude better stoichiometric control, lower temperature requirements, and chemically uniform products. Several phosphate sources were investigated, including phosphoric acid, pyrophosphoric acid, and potassium pyrophosphate. The resulting indium-doped tin pyrophosphates had good proton conductivity over a wide temperature range with no humidification.

  12. Pyrophosphate-Fueled Na+ and H+ Transport in Prokaryotes

    PubMed Central

    Malinen, Anssi M.; Luoto, Heidi H.

    2013-01-01

    SUMMARY In its early history, life appeared to depend on pyrophosphate rather than ATP as the source of energy. Ancient membrane pyrophosphatases that couple pyrophosphate hydrolysis to active H+ transport across biological membranes (H+-pyrophosphatases) have long been known in prokaryotes, plants, and protists. Recent studies have identified two evolutionarily related and widespread prokaryotic relics that can pump Na+ (Na+-pyrophosphatase) or both Na+ and H+ (Na+,H+-pyrophosphatase). Both these transporters require Na+ for pyrophosphate hydrolysis and are further activated by K+. The determination of the three-dimensional structures of H+- and Na+-pyrophosphatases has been another recent breakthrough in the studies of these cation pumps. Structural and functional studies have highlighted the major determinants of the cation specificities of membrane pyrophosphatases and their potential use in constructing transgenic stress-resistant organisms. PMID:23699258

  13. Compositional changes of a dicalcium phosphate dihydrate cement after implantation in sheep.

    PubMed

    Bohner, M; Theiss, F; Apelt, D; Hirsiger, W; Houriet, R; Rizzoli, G; Gnos, E; Frei, C; Auer, J A; von Rechenberg, B

    2003-09-01

    A hydraulic calcium phosphate cement having dicalcium phosphate dihydrate (DCPD) as end-product of the setting reaction was implanted in a cylindrical defect in the diaphysis of sheep for up to 6 months. The composition of the cement was investigated as a function of time. After setting, the cement composition consisted essentially of a mixture of DCPD and beta-tricalcium phosphate (beta-TCP). In the first few weeks of implantation, the edges of the cement samples became depleted in DCPD, suggesting a selective dissolution of DCPD, possibly due to low pH conditions. The cement resorption at this stage was high. After 8 weeks, the resorption rate slowed down. Simultaneously, a change of the color and density of the cement center was observed. These changes were due to the conversion of DCPD into a poorly crystalline apatite. Precipitation started after 6-8 weeks and progressed rapidly. At 9 weeks, the colored central zone reached its maximal size. The fraction of beta-TCP in the cement was constant at all time. Therefore, this study demonstrates that the resorption rate of DCPD cement is more pronounced as long as DCPD is not transformed in vivo.

  14. Comparative intraoral tolerance of sodium hexametaphosphate and pyrophosphate antitartar dentifrices.

    PubMed

    Gerlach, Robert W

    2002-01-01

    A randomized, controlled clinical trial was conducted to assess the relative acute oral tolerance of a novel, dual-phase, tartar control dentifrice containing 7.0% sodium hexametaphosphate (5% hexametaphosphate anion). In this 4-day study, 159 healthy adult volunteers were randomly assigned to the experimental sodium hexametaphosphate dentifrice or a marketed, single-phase, antitartar dentifrice control containing 5.0% ionic pyrophosphate. A detailed oral soft tissue examination and interview were conducted each day by blinded evaluators to elicit clinical signs and symptoms associated with ad libitum use of the assigned dentifrice. A total of 24 subjects (15% of the study population) had new symptoms/signs after baseline. By treatment, 9% of subjects in the sodium hexametaphosphate group had new findings, compared to 21% of subjects in the pyrophosphate group, and these groups differed statistically (p < 0.03, two-sided) with respect to occurrence. In addition, onset, severity, duration and clinical presentation were generally milder in the sodium hexametaphosphate group compared to the pyrophosphate control. Only one subject (in the pyrophosphate group) discontinued treatment early due to oral intolerance. In this study, the 7.0% sodium hexametaphosphate dentifrice was well-tolerated, with significantly fewer symptoms and superior overall tolerance compared to the marketed antitartar dentifrice control.

  15. 21 CFR 582.5306 - Ferric sodium pyrophosphate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 6 2014-04-01 2014-04-01 false Ferric sodium pyrophosphate. 582.5306 Section 582.5306 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients...

  16. 21 CFR 582.5306 - Ferric sodium pyrophosphate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 6 2012-04-01 2012-04-01 false Ferric sodium pyrophosphate. 582.5306 Section 582.5306 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients...

  17. 21 CFR 582.5306 - Ferric sodium pyrophosphate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 6 2013-04-01 2013-04-01 false Ferric sodium pyrophosphate. 582.5306 Section 582.5306 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients...

  18. 21 CFR 582.5306 - Ferric sodium pyrophosphate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Ferric sodium pyrophosphate. 582.5306 Section 582.5306 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Nutrients...

  19. Amyloid goiter: preoperative scintigraphic diagnosis using Tc-99m pyrophosphate

    SciTech Connect

    Lee, V.W.; Rubinow, A.; Pehrson, J.; Skinner, M.; Cohen, A.S.

    1984-04-01

    Amyloid goiter is a rare clinical entity. The diagnosis is rarely made preoperatively because clinical and laboratory findings are nonspecific. The authors report two cases of amyloid goiter in whom the diagnosis was made preoperatively using Tc-99m pyrophosphate scintigraphy.

  20. Fabrication of dicalcium phosphate dihydrate-coated β-TCP granules and evaluation of their osteoconductivity using experimental rats.

    PubMed

    Shariff, Khairul Anuar; Tsuru, Kanji; Ishikawa, Kunio

    2017-06-01

    β-Tricalcium phosphate (β-TCP) has attracted much attention as an artificial bone substitute owing to its biocompatibility and osteoconductivity. In this study, osteoconductivity of β-TCP bone substitute was enhanced without using growth factors or cells. Dicalcium phosphate dihydrate (DCPD), which is known to possess the highest solubility among calcium phosphates, was coated on β-TCP granules by exposing their surface with acidic calcium phosphate solution. The amount of coated DCPD was regulated by changing the reaction time between β-TCP granules and acidic calcium phosphate solution. Histomorphometry analysis obtained from histological results revealed that the approximately 10mol% DCPD-coated β-TCP granules showed the largest new bone formation compared to DCPD-free β-TCP granules, approximately 2.5mol% DCPD-coated β-TCP granules, or approximately 27mol% DCPD-coated β-TCP granules after 2 and 4weeks of implantation. Based on this finding, we demonstrate that the osteoconductivity of β-TCP granules could be improved by coating their surface with an appropriate amount of DCPD. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Dissolution behaviour of ferric pyrophosphate and its mixtures with soluble pyrophosphates: Potential strategy for increasing iron bioavailability.

    PubMed

    Tian, Tian; Blanco, Elena; Smoukov, Stoyan K; Velev, Orlin D; Velikov, Krassimir P

    2016-10-01

    Ferric pyrophosphate (FePP) is a widely used iron source in food fortification and in nutritional supplements, due to its white colour, that is very uncommon for insoluble Fe salts. Although its dissolution is an important determinant of Fe adsorption in human body, the solubility characteristics of FePP are complex and not well understood. This report is a study on the solubility of FePP as a function of pH and excess of pyrophosphate ions. FePP powder is sparingly soluble in the pH range of 3-6 but slightly soluble at pH<2 and pH>8. In the presence of pyrophosphate ions the solubility of FePP strongly increases at pH 5-8.5 due to formation a soluble complex between Fe(III) and pyrophosphate ions, which leads to an 8-10-fold increase in the total ionic iron concentration. This finding is beneficial for enhancing iron bioavailability, which important for the design of fortified food, beverages, and nutraceutical products.

  2. Lisinopril dihydrate: single-crystal x-ray structure and physicochemical characterization of derived solid forms.

    PubMed

    Sorrenti, Milena; Catenacci, Laura; Cruickshank, Dyanne L; Caira, Mino R

    2013-10-01

    Screening for new solid forms of the antihypertensive lisinopril was performed by recrystallization of the commercial form, lisinopril dihydrate, from various solvents and by exposing the product of its dehydration to a series of vapors under controlled conditions. Modifications other than the dihydrate encountered in the study included new anhydrous and amorphous forms, with intrinsic dissolution rates significantly greater than that of the dihydrate. Further physicochemical characterization included constant and programmed temperature powder X-ray diffraction, differential scanning calorimetry, thermogravimetry, and Fourier transform infrared spectroscopy. In the course of this study, the single-crystal X-ray structure of lisinopril dihydrate, [a = 14.550(2), b = 5.8917(8), c = 14.238(2) Å, β = 112.832(3)° at T = 173(2) K, space group P21 , Z = 2], was determined for the first time, revealing its double zwitterionic character in the solid state. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association.

  3. Interpretation of dissolution kinetics of dicalcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Zhang, Jingwu; Nancollas, George H.

    1992-11-01

    The kinetics of dissolution of dicalcium phosphate dihydrate (DCPD) has been investigated at physiological temperature 37°C using the constant composition method. In order to calculate the undersaturation, the solubility product was determined at the ionic strength of the dissolution experiments. The rates were sensitive to the solution hydrodynamics at high driving forces and under mild agitation, suggesting that volume diffusion provides a substantial resistance to dissolution. However, surface processes became rate determining at low undersaturations as reflected by the insensitivity of rates to changes of stirring speed and the relatively high activation energy. The dependence of the rate upon the relative undersaturation was analyzed using recent crystal growth theories. Though not necessarily rate determining, surface diffusion appeared to be involved in transporting dissolved ions into the solution phase.

  4. Thiamine pyrophosphate riboswitches are targets for the antimicrobial compound pyrithiamine.

    PubMed

    Sudarsan, Narasimhan; Cohen-Chalamish, Smadar; Nakamura, Shingo; Emilsson, Gail Mitchell; Breaker, Ronald R

    2005-12-01

    Thiamine metabolism genes are regulated in numerous bacteria by a riboswitch class that binds the coenzyme thiamine pyrophosphate (TPP). We demonstrate that the antimicrobial action of the thiamine analog pyrithiamine (PT) is mediated by interaction with TPP riboswitches in bacteria and fungi. For example, pyrithiamine pyrophosphate (PTPP) binds the TPP riboswitch controlling the tenA operon in Bacillus subtilis. Expression of a TPP riboswitch-regulated reporter gene is reduced in transgenic B. subtilis or Escherichia coli when grown in the presence of thiamine or PT, while mutant riboswitches in these organisms are unresponsive to these ligands. Bacteria selected for PT resistance bear specific mutations that disrupt ligand binding to TPP riboswitches and derepress certain TPP metabolic genes. Our findings demonstrate that riboswitches can serve as antimicrobial drug targets and expand our understanding of thiamine metabolism in bacteria.

  5. Pyrophosphate-condensing activity linked to nucleic acid synthesis.

    PubMed Central

    Volloch, V Z; Rits, S; Tumerman, L

    1979-01-01

    In some preparations of DNA dependent RNA polymerase a new enzymatic activity has been found which catalyzes the condensation of two pyrophosphate molecules, liberated in the process of RNA synthesis, to one molecule of orthophosphate and one molecule of Mg (or Mn) - chelate complex with trimetaphosphate. This activity can also cooperate with DNA-polymerase, on condition that both enzymes originate from the same cells. These results point to two general conclusions. First, energy is conserved in the overall process of nucleic acid synthesis and turnover, so that the process does not require an energy influx from the cell's general resources. Second, the synthesis of nucleic acids is catalyzed by a complex enzyme system which contains at least two separate enzymes, one responsible for nucleic acid polymerization and the other for energy conservation via pyrophosphate condensation. Images PMID:88040

  6. Tc-99m pyrophosphate myocardial scanning in Chagas' disease

    SciTech Connect

    Goncalves da Rocha, A.F.; Meguerian, B.A.; Harbert, J.C.

    1981-04-01

    Chagas' disease is a serious protozoan infection affecting up to 20% of populations in some endemic areas. Myocarditis and cardiomyopathy occur in 50% of patients who go on to develop chronic Chagas's disease. We have studied a patient with no overt cardiac symptoms who revealed intense myocardial uptake of Tc-99m pyrophosphate. The significance of this finding in relation to early detection and progress of therapy is explored.

  7. Tc-99m pyrophosphate myocardial scanning in Chagas' disease

    SciTech Connect

    da Rocha, A.F.; Meguerian, B.A.; Harbert, J.C.

    1981-04-01

    Chagas' disease is a serious protozoan infection affecting up to 20% of populations in some endemic areas. Myocarditis and cardiomyopathy occur in 50% of patients who go on to develop chronic Chagas' disease. We have studied a patient with no overt cardiac symptoms who revealed intense myocardial uptake of Tc-99m pyrophosphate. The significance of this finding in relation to early detection and progress of therapy is explored.

  8. The coenzyme thiamine pyrophosphate inhibits the self-splicing of the group I intron.

    PubMed

    Ahn, Sung Joon; Park, In Kook

    2003-02-01

    Effects of the coenzyme thiamine pyrophosphate and its analogs on the inhibition of self-splicing of primary transcripts of the phage T4 thymidylate synthase gene (td) were investigated. Of all compounds tested, the coenzyme thiamine pyrophosphate was the most potent inhibitor and the order of inhibitory efficiency for compounds tested was as follows: thiamine pyrophosphate>thiamine monophosphate>thiamine>thiochrome. Increasing guanosine concentration overcame the suppression of self-splicing by thiamine pyrophosphate close to the level of normal splicing. Kinetic analysis demonstrated that thiamine pyrophosphate acts as a competitive inhibitor for the td intron RNA with a Ki of 2.2mM. The splicing specificity inhibition by thiamine pyrophosphate is predominantly due to changes in Km.

  9. "Fabrication of arbitrarily shaped carbonate apatite foam based on the interlocking process of dicalcium hydrogen phosphate dihydrate".

    PubMed

    Sugiura, Yuki; Tsuru, Kanji; Ishikawa, Kunio

    2017-08-01

    Carbonate apatite (CO3Ap) foam with an interconnected porous structure is highly attractive as a scaffold for bone replacement. In this study, arbitrarily shaped CO3Ap foam was formed from α-tricalcium phosphate (α-TCP) foam granules via a two-step process involving treatment with acidic calcium phosphate solution followed by hydrothermal treatment with NaHCO3. The treatment with acidic calcium phosphate solution, which is key to fabricating arbitrarily shaped CO3Ap foam, enables dicalcium hydrogen phosphate dihydrate (DCPD) crystals to form on the α-TCP foam granules. The generated DCPD crystals cause the α-TCP granules to interlock with each other, inducing an α-TCP/DCPD foam. The interlocking structure containing DCPD crystals can survive hydrothermal treatment with NaHCO3. The arbitrarily shaped CO3Ap foam was fabricated from the α-TCP/DCPD foam via hydrothermal treatment at 200 °C for 24 h in the presence of a large amount of NaHCO3.

  10. Fabrication of interconnected porous calcite by bridging calcite granules with dicalcium phosphate dihydrate and their histological evaluation.

    PubMed

    Ishikawa, Kunio; Koga, Noriko; Tsuru, Kanji; Takahashi, Ichiro

    2016-03-01

    Interconnected porous calcite has attracted attention as an artificial bone replacement material and as a precursor for the fabrication of carbonate apatite, which is also an artificial bone replacement material. In this study, calcite granules were exposed to acidic calcium phosphate solution, and the feasibility of fabricating interconnected porous calcite using this process was evaluated. No setting reaction was observed under the nonloading condition. In contrast, under loading conditions, calcite granules were bridged with dicalcium phosphate dihydrate crystals, and the calcite granules set into interconnected porous calcite foam. When applied 0.4 MPa of loading pressure during sample preparation, compressive strength of the obtained interconnected porous calcite was ∼1.5 MPa. The exposure of the calcite granules to acidic calcium phosphate solution under loading conditions was the key for the setting reaction to occur. This is because calcite granules cannot contact one another under the nonloading condition because of bubble formation on the surfaces of the calcite granules. The interconnected porous calcite revealed excellent tissue response, and new bone was able to penetrate into the porous calcite 2 weeks after implantation. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A 104A: 652-658, 2016. © 2015 Wiley Periodicals, Inc.

  11. Poly(Propylene Fumarate) Reinforced Dicalcium Phosphate Dihydrate Cement Composites for Bone Tissue Engineering

    PubMed Central

    Alge, Daniel L.; Bennet, Jeffrey; Treasure, Trevor; Voytik-Harbin, Sherry; Goebel, W. Scott; Chu, Tien-Min Gabriel

    2012-01-01

    Calcium phosphate cements have many desirable properties for bone tissue engineering, including osteoconductivity, resorbability, and amenability to rapid prototyping based methods for scaffold fabrication. In this study, we show that dicalcium phosphate dihydrate (DCPD) cements, which are highly resorbable but also inherently weak and brittle, can be reinforced with poly(propylene fumarate) (PPF) to produce strong composites with mechanical properties suitable for bone tissue engineering. Characterization of DCPD-PPF composites revealed significant improvements in mechanical properties for cements with a 1.0 powder to liquid ratio. Compared to non-reinforced controls, flexural strength improved from 1.80 ± 0.19 MPa to 16.14 ± 1.70 MPa, flexural modulus increased from 1073.01 ± 158.40 MPa to 1303.91 ± 110.41 MPa, maximum displacement during testing increased from 0.11 ± 0.04 mm to 0.51 ± 0.09 mm, and work of fracture improved from 2.74 ± 0.78 J/m2 to 249.21 ± 81.64 J/m2. To demonstrate the utility of our approach for scaffold fabrication, 3D macroporous scaffolds were prepared with rapid prototyping technology. Compressive testing revealed that PPF reinforcement increased scaffold strength from 0.31 ± 0.06 MPa to 7.48 ± 0.77 MPa. Finally, 3D PPF-DCPD scaffolds were implanted into calvarial defects in rabbits for 6 weeks. Although the addition of mesenchymal stem cells to the scaffolds did not significantly improve the extent of regeneration, numerous bone nodules with active osteoblasts were observed within the scaffold pores, especially in the peripheral regions. Overall, the results of this study suggest that PPF-DCPD composites may be promising scaffold materials for bone tissue engineering. PMID:22489012

  12. Poly(propylene fumarate) reinforced dicalcium phosphate dihydrate cement composites for bone tissue engineering.

    PubMed

    Alge, Daniel L; Bennett, Jeffrey; Treasure, Trevor; Voytik-Harbin, Sherry; Goebel, W Scott; Chu, Tien-Min Gabriel

    2012-07-01

    Calcium phosphate cements have many desirable properties for bone tissue engineering, including osteoconductivity, resorbability, and amenability to rapid prototyping-based methods for scaffold fabrication. In this study, we show that dicalcium phosphate dihydrate (DCPD) cements, which are highly resorbable but also inherently weak and brittle, can be reinforced with poly(propylene fumarate) (PPF) to produce strong composites with mechanical properties suitable for bone tissue engineering. Characterization of DCPD-PPF composites revealed significant improvements in mechanical properties for cements with a 1.0 powder to liquid ratio. Compared with nonreinforced controls, flexural strength improved from 1.80 ± 0.19 MPa to 16.14 ± 1.70 MPa, flexural modulus increased from 1073.01 ± 158.40 MPa to 1303.91 ± 110.41 MPa, maximum displacement during testing increased from 0.11 ± 0.04 mm to 0.51 ± 0.09 mm, and work of fracture improved from 2.74 ± 0.78 J/m(2) to 249.21 ± 81.64 J/m(2) . To demonstrate the utility of our approach for scaffold fabrication, 3D macroporous scaffolds were prepared with rapid prototyping technology. Compressive testing revealed that PPF reinforcement increased scaffold strength from 0.31 ± 0.06 MPa to 7.48 ± 0.77 MPa. Finally, 3D PPF-DCPD scaffolds were implanted into calvarial defects in rabbits for 6 weeks. Although the addition of mesenchymal stem cells to the scaffolds did not significantly improve the extent of regeneration, numerous bone nodules with active osteoblasts were observed within the scaffold pores, especially in the peripheral regions. Overall, the results of this study suggest that PPF-DCPD composites may be promising scaffold materials for bone tissue engineering.

  13. SEPARATION OF PLUTONIUM IONS FROM SOLUTION BY ADSORPTION ON ZIRCONIUM PYROPHOSPHATE

    DOEpatents

    Stoughton, R.W.

    1961-01-31

    A method is given for separating plutonium in its reduced, phosphate- insoluble state from other substances. It involves contacting a solution containing the plutonium with granular zirconium pyrophosphate.

  14. Dimethylallyl pyrophosphate is not the committed precursor of isopentenyl pyrophosphate during terpenoid biosynthesis from 1-deoxyxylulose in higher plants.

    PubMed

    Arigoni, D; Eisenreich, W; Latzel, C; Sagner, S; Radykewicz, T; Zenk, M H; Bacher, A

    1999-02-16

    Cell cultures of Catharanthus roseus were supplied with [2-13C, 3-2H]-deoxyxylulose or [2-13C,4-2H]1-deoxyxylulose. Lutein and chlorophylls were isolated from the cell mass, and hydrolysis of the chlorophyll mixtures afforded phytol. Isotope labeling patterns of phytol and lutein were determined by 2H NMR and 1H,2H-decoupled 13C NMR. From the data it must be concluded that the deuterium atom in position 3 of deoxyxylulose was incorporated into both isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate with a rate of 75% (with respect to the internal 13C label). The detected stereochemical signature implies that the label is located preferentially in the (E)-hydrogen atom of IPP. This preferential labeling, in turn, rules out dimethylallyl pyrophosphate as the compulsory precursor of IPP. In the experiment with [2-13C, 4-2H]1-deoxyxylulose, the 13C label was efficiently transferred to the terpenoids whereas the 2H label was completely washed out, most probably after IPP formation as a consequence of the isomerization and elongation process. In addition, the data cast light on the stereochemical course of the dehydrogenation and cyclization steps involved in the biosynthesis of lutein.

  15. Calcium supplements

    MedlinePlus

    ... TYPES OF CALCIUM SUPPLEMENTS Forms of calcium include: Calcium carbonate: Over-the-counter (OTC) antacid products, such as Tums and Rolaids, contain calcium carbonate. These sources of calcium do not cost much. ...

  16. Crystal growth of calcium phosphates - epitaxial considerations

    NASA Astrophysics Data System (ADS)

    Koutsoukos, P. G.; Nancollas, G. H.

    1981-05-01

    The growth of one crystalline phase on the surface of another that offers a good crystal lattice match, may be important in environmental, physiological and pathological mineralization processes. The epitaxial relationships and kinetics of growth of hydroxyapatite on crystals of dicalcium phosphate dihydrate, calcium fluoride and calcite have been studied at sustained low supersaturation with respect to hydroxyapatite. At the very low supersaturations, the crystallization of hydroxyapatite takes place without the formation of precursor phases. The experimental results are in agreement with theoretical predictions for epitaxial growth, while the kinetics of hydroxyapatite crystallization on the foreign substrates is the same as that for the growth of hydroxyapatite on synthetic hydroxyapatite crystals.

  17. Calcium phosphate scaffold from biogenic calcium carbonate by fast ambient condition reactions

    NASA Astrophysics Data System (ADS)

    Dutta, Abhishek; Fermani, Simona; Arjun Tekalur, Srinivasan; Vanderberg, Abigail; Falini, Giuseppe

    2011-12-01

    Calcium phosphate biogenic materials are biocompatible and promote bioactivity and osteoconductivity, which implies their natural affinity and tendency to bond directly to bones subsequently replacing the host bone after implantation owing to its biodegradability. Calcium hydrogen phosphate dihydrate, CaHPO 4·2H 2O, is known to be a nucleation precursor, in aqueous solutions, for apatitic calcium phosphates and, hence, a potential starting material for bone substitutes. Numerous approaches, via hydrothermal and ambient synthetic routes, have been used to produce calcium phosphate from biogenic calcium carbonate, taking advantage of the peculiar architecture and composition of the latter. In this article, the lamellar region of the cuttlefish bone ( Sepia officinalis) was used as a framework for the organized deposition of calcium phosphate crystals, at ambient conditions via a fast procedure involving an amorphous calcium carbonate intermediate, and ending with a conversion to calcium phosphate and a fixation procedure, thereby resulting in direct conversion of biogenic calcium carbonate into calcium phosphates at ambient conditions from the scale of months to hours.

  18. Effect of waters of crystallization on terahertz spectra: anhydrous oxalic acid and its dihydrate.

    PubMed

    King, Matthew D; Korter, Timothy M

    2010-07-08

    Oxalic acid and oxalic acid dihydrate were studied using terahertz spectroscopy and solid-state density functional theory (DFT) in the spectral range 10-100 cm(-1). The size of the oxalic acid molecule and its limited internal degrees of freedom make it ideal for evaluating the performance of computational methods for the structural and dynamical simulation of strongly hydrogen-bonded solids. Calculations of the solid-state structures and terahertz spectra of oxalic acid and oxalic acid dihydrate were performed using the hybrid B3LYP and B3PW91 and the nonhybrid BLYP and PW91 density functionals employing the 6-311G(2d,2p) basis set. When these simulations were compared to the experimental spectra of the oxalic acid solids, a constant overprediction of the dihydrate frequencies was observed in contrast to the results of the anhydrous system. This change in behavior is connected to the nature of the vibrational motions being accessed. The primary molecular motion contributions to the terahertz vibrations of oxalic acid dihydrate were found to originate in the external motions of the cocrystallized H(2)O molecules. The observed overestimation of the vibrational energies in the simulated terahertz spectra is attributed to increased anharmonicity of the vibrational motions in the dihydrate system versus the anhydrous, resulting from weaker hydrogen bonding through the networked water molecules.

  19. Structure and spectral properties of L-histidinium dipicrate dihydrate.

    PubMed

    Sethuram, M; Dhandapani, M; Sethu Raman, M; Amirthaganesan, G; Senthilkumar, K

    2014-01-24

    Non-linear optical active L-histidinium dipicrate dihydrate (LHDD) single crystals were grown by slow evaporation method. The Fourier transform FT-IR, FT-Raman, (1)H and (13)C NMR spectra of the crystal have been recorded and analysed. The spectral analyses confirm the formation of the compound and the stoichiometry. The geometry and spectral characteristics were examined using the density functional theory (DFT) method, B3LYP with 6-31G(d,p) basis set. The first-order hyperpolarisability, energies of frontier molecular orbitals and the Mulliken population analysis were also calculated. The HOMO-LUMO energy gap value 2.73 eV indicates the charge transfer from picrate to histidinium through the hydrogen bond. The second-order NLO properties of the molecule are studied by Kurtz-Perry powder technique. SHG efficiency of the compound is nearly 2.5 times greater than KDP. Theoretical calculations indicate hyperpolarisability of LHDD is 39 times greater than urea. The results show that the title molecule can be used for opto-electronic applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Dissolution kinetics of dicalcium phosphate dihydrate under pseudophysiological conditions

    NASA Astrophysics Data System (ADS)

    Kanzaki, Noriko; Onuma, Kazuo; Treboux, Gabin; Ito, Atsuo

    2002-02-01

    Dissolution kinetics of the (0 1 0) face of dicalcium phosphate dihydrate (DCPD) and the transformation of DCPD to hydroxyapatite were observed using in situ atomic force microscopy. Step velocities for [2 0 1]- and [1 0 1]'-oriented steps were measured in pseudophysiological solutions. The latter step was inclined 5-7° toward [0 0 1] from [1 0 1] directions. The step kinetic coefficients calculated for the [2 0 1] and [1 0 1]' steps were, respectively, 0.70×10 -4 and 1.43×10 -4 m/s, values comparable to those for soluble inorganic crystals. The solubility product for DCPD accurately estimated from the relation between the step velocity and the undersaturation was (4.13±0.1)×10 -7, a value 1.6 times larger than that previously reported. It was shown that the precipitation of hydroxyapatite occurred after the dissolution of DCPD, and no evidence of direct structural transformation from DCPD to hydroxyapatite was observed. This indicates that DCPD acts as a simple heterogeneous growth center for hydroxyapatite without requiring any structural resemblance.

  1. Protein pyrophosphorylation by inositol pyrophosphates is a posttranslational event

    PubMed Central

    Bhandari, Rashna; Saiardi, Adolfo; Ahmadibeni, Yousef; Snowman, Adele M.; Resnick, Adam C.; Kristiansen, Troels Z.; Molina, Henrik; Pandey, Akhilesh; Werner, J. Kent; Juluri, Krishna R.; Xu, Yong; Prestwich, Glenn D.; Parang, Keykavous; Snyder, Solomon H.

    2007-01-01

    In a previous study, we showed that the inositol pyrophosphate diphosphoinositol pentakisphosphate (IP7) physiologically phosphorylates mammalian and yeast proteins. We now report that this phosphate transfer reflects pyrophosphorylation. Thus, proteins must be prephosphorylated by ATP to prime them for IP7 phosphorylation. IP7 phosphorylates synthetic phosphopeptides but not if their phosphates have been masked by methylation or pyrophosphorylation. Moreover, IP7 phosphorylated peptides are more acid-labile and more resistant to phosphatases than ATP phosphorylated peptides, indicating a different type of phosphate bond. Pyrophosphorylation may represent a novel mode of signaling to proteins. PMID:17873058

  2. Polymerization of the cyclic pyrophosphates of nucleosides and their analogues

    NASA Technical Reports Server (NTRS)

    Tohidi, Mahrokh; Orgel, Leslie E.

    1990-01-01

    When 2-prime-deoxythymidine 3-prime, 5-prime-cyclic diphosphate, or the cyclic pyrophosphates of the acyclic nucleoside analogs II and IV are heated to 65-85 C in the presence of imidazole, oligomers with lengths up to 20-30 are formed in excellent yield. This reaction provides a useful source of oligomers for use as templates in aqueous condensation reactions. In the absence of evidence to the contrary, it is assumed that the oligomers are atactic. The potential significance of this reaction in prebiotic chemistry is discussed.

  3. Polymerization of the cyclic pyrophosphates of nucleosides and their analogues

    NASA Technical Reports Server (NTRS)

    Tohidi, Mahrokh; Orgel, Leslie E.

    1990-01-01

    When 2-prime-deoxythymidine 3-prime, 5-prime-cyclic diphosphate, or the cyclic pyrophosphates of the acyclic nucleoside analogs II and IV are heated to 65-85 C in the presence of imidazole, oligomers with lengths up to 20-30 are formed in excellent yield. This reaction provides a useful source of oligomers for use as templates in aqueous condensation reactions. In the absence of evidence to the contrary, it is assumed that the oligomers are atactic. The potential significance of this reaction in prebiotic chemistry is discussed.

  4. Distribution of Endogenous Farnesyl Pyrophosphate and Four Species of Lysophosphatidic Acid in Rodent Brain

    PubMed Central

    Lee, Sung Ha; Raboune, Siham; Walker, J. Michael; Bradshaw, Heather B.

    2010-01-01

    Lysophosphatidic acid (LPA) is the umbrella term for lipid signaling molecules that share structural homology and activate the family of LPA receptors. Farnesyl Pyrophosphate (FPP) is commonly known as an intermediate in the synthesis of steroid hormones; however, its function as a signaling lipid is beginning to be explored. FPP was recently shown to an activator of the G-protein coupled receptor 92 (also known as LPA5) of the calcium channel TRPV3. The LPA receptors (including GPR92) are associated with the signal transduction of noxious stimuli, however, very little is known about the distribution of their signaling ligands (LPAs and FPP) in the brain. Here, using HPLC/MS/MS, we developed extraction and analytical methods for measuring levels of FPP and 4 species of LPA (palmitoyl, stearoyl, oleoyl and arachidonoyl-sn-glycerol-3 phosphate) in rodent brain. Relative distributions of each of the five compounds was significantly different across the brain suggesting divergent functionality for each as signaling molecules based on where and how much of each is being produced. Brainstem, midbrain, and thalamus contained the highest levels measured for each compound, though none in the same ratios while relatively small amounts were produced in cortex and cerebellum. These data provide a framework for investigations into functional relationships of these lipid ligands in specific brain areas, many of which are associated with the perception of pain. PMID:21152313

  5. Novel phosphate-activated macrophages prevent ectopic calcification by increasing extracellular ATP and pyrophosphate

    PubMed Central

    Villa-Bellosta, Ricardo; Hamczyk, Magda R.; Andrés, Vicente

    2017-01-01

    Purpose Phosphorus is an essential nutrient involved in many pathobiological processes. Less than 1% of phosphorus is found in extracellular fluids as inorganic phosphate ion (Pi) in solution. High serum Pi level promotes ectopic calcification in many tissues, including blood vessels. Here, we studied the effect of elevated Pi concentration on macrophage polarization and calcification. Macrophages, present in virtually all tissues, play key roles in health and disease and display remarkable plasticity, being able to change their physiology in response to environmental cues. Methods and results High-throughput transcriptomic analysis and functional studies demonstrated that Pi induces unpolarized macrophages to adopt a phenotype closely resembling that of alternatively-activated M2 macrophages, as revealed by arginine hydrolysis and energetic and antioxidant profiles. Pi-induced macrophages showed an anti-calcifying action mediated by increased availability of extracellular ATP and pyrophosphate. Conclusion We conclude that the ability of Pi-activated macrophages to prevent calcium-phosphate deposition is a compensatory mechanism protecting tissues from hyperphosphatemia-induced pathologic calcification. PMID:28362852

  6. Alizarin Red S for Online Pyrophosphate Detection Identified by a Rapid Screening Method

    PubMed Central

    Fischbach, Jens; Loh, Qiuting; Bier, Frank F.; Lim, Theam Soon; Frohme, Marcus; Glökler, Jörn

    2017-01-01

    We identified Alizarin Red S and other well known fluorescent dyes useful for the online detection of pyrophosphate in enzymatic assays, including the loop mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assays. An iterative screening was used for a selected set of compounds to first secure enzyme compatibility, evaluate inorganic pyrophosphate sensitivity in the presence of manganese as quencher and optimize conditions for an online detection. Of the selected dyes, the inexpensive alizarin red S was found to selectively detect pyrophosphate under LAMP and PCR conditions and is superior with respect to its defined red-shifted spectrum, long shelf life and low toxicity. In addition, the newly identified properties may also be useful in other enzymatic assays which do not generate nucleic acids but are based on inorganic pyrophosphate. Finally, we propose that our screening method may provide a blueprint for rapid screening of compounds for detecting inorganic pyrophosphate. PMID:28338022

  7. Nitrogen-containing bisphosphonates inhibit isopentenyl pyrophosphate isomerase/farnesyl pyrophosphate synthase activity with relative potencies corresponding to their antiresorptive potencies in vitro and in vivo.

    PubMed

    van Beek, E; Pieterman, E; Cohen, L; Löwik, C; Papapoulos, S

    1999-02-16

    Bisphosphonates, synthetic compounds which suppress bone resorption, are used in the treatment of skeletal disorders. Their mode of action and intracellular targets have not yet been identified. Recent evidence suggested that enzymes of the mevalonate pathway are the potential targets. In this study, we examined the effect of four potent nitrogen (N)-containing bisphosphonates, clodronate and NH2-olpadronate, an inactive analogue of olpadronate, on isopentenyl pyrophosphate isomerase/farnesyl pyrophosphate synthase, geranylgeranyl pyrophosphate synthase, and protein geranylgeranyl transferase I activity. We found that all N-containing bisphosphonates inhibited isopentenyl pyrophosphate isomerase/farnesyl pyrophosphate synthase activity dose dependently with relative potencies corresponding to their antiresorptive potencies in vitro and in vivo, whereas clodronate and NH2-olpadronate had no effect. Furthermore, none of the bisphosphonates tested affected geranylgeranyl pyrophosphate synthase or geranylgeranyl transferase I activity. Our study reveals for the first time the intracellular target of N-containing bisphosphonates and supports the view that all bisphosphonates do not share the same molecular mechanism of action. Copyright 1999 Academic Press.

  8. Biochemical characterization of the Mycobacterium tuberculosis phosphoribosyl-1-pyrophosphate synthetase

    PubMed Central

    Alderwick, Luke J; Lloyd, Georgina S; Lloyd, Adrian J; Lovering, Andrew L; Eggeling, Lothar; Besra, Gurdyal S

    2011-01-01

    Mycobacterium tuberculosis arabinogalactan (AG) is an essential cell wall component. It provides a molecular framework serving to connect peptidoglycan to the outer mycolic acid layer. The biosynthesis of the arabinan domains of AG and lipoarabinomannan (LAM) occurs via a combination of membrane bound arabinofuranosyltransferases, all of which utilize decaprenol-1-monophosphorabinose as a substrate. The source of arabinose ultimately destined for deposition into cell wall AG or LAM originates exclusively from phosphoribosyl-1-pyrophosphate (pRpp), a central metabolite which is also required for other essential metabolic processes, such as de novo purine and pyrimidine biosyntheses. In M. tuberculosis, a single pRpp synthetase enzyme (Mt-PrsA) is solely responsible for the generation of pRpp, by catalyzing the transfer of pyrophosphate from ATP to the C1 hydroxyl position of ribose-5-phosphate. Here, we report a detailed biochemical and biophysical study of Mt-PrsA, which exhibits the most rapid enzyme kinetics reported for a pRpp synthetase. PMID:21045009

  9. Label-Free Pyrophosphate Recognition with Functionalized Asymmetric Nanopores.

    PubMed

    Ali, Mubarak; Ahmed, Ishtiaq; Ramirez, Patricio; Nasir, Saima; Niemeyer, Christof M; Mafe, Salvador; Ensinger, Wolfgang

    2016-04-01

    The label-free detection of pyrophosphate (PPi) anions with a nanofluidic sensing device based on asymmetric nanopores is demonstrated. The pore surface is functionalized with zinc complexes based on two di(2-picolyl)amine [bis(DPA)] moieties using carbodiimide coupling chemistry. The complexation of zinc (Zn(2+) ) ion is achieved by exposing the modified pore to a solution of zinc chloride to form bis(Zn(2+) -DPA) complexes. The chemical functionalization is demonstrated by recording the changes in the observed current-voltage (I-V) curves before and after pore modification. The bis(Zn(2+) -DPA) complexes on the pore walls serve as recognition sites for pyrophosphate anion. The experimental results show that the proposed nanofluidic sensor has the ability to sense picomolar concentrations of PPi anion in the surrounding environment. On the contrary, it does not respond to other phosphate anions, including monohydrogen phosphate, dihydrogen phosphate, adenosine monophosphate, adenosine diphosphate, and adenosine triphosphate. The experimental results are described theoretically by using a model based on the Poisson-Nernst-Planck equations. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Subcellular pyrophosphate metabolism in developing tubers of potato (Solanum tuberosum).

    PubMed

    Farré, Eva M; Tech, Susanne; Trethewey, Richard N; Fernie, Alisdair R; Willmitzer, Lothar

    2006-09-01

    PPi has previously been implicated specifically in the co-ordination of the sucrose-starch transition and in the broader context of its role as co-factor in heterotrophic plant metabolism. In order to assess the compartmentation of pyrophosphate (PPi) metabolism in the potato tuber we analysed the effect of expressing a bacterial pyrophosphatase in the amyloplast of wild type tubers or in the cytosol or amyloplast of invertase-expressing tubers. The second and third approaches were adopted since we have previously characterized the invertase expressing lines to both exhibit highly altered sucrose metabolism and to contain elevated levels of PPi (Farré et al. (2000a) Plant Physiol 123:681) and therefore this background rendered questions concerning the level of communication between the plastidic and cytosolic pyrophosphate pools relatively facile. In this study we observed that the increase in PPi in the invertase expressing lines was mainly confined to the cytosol. Accordingly, the expression of a bacterial pyrophosphatase in the plastid of either wild type or invertase-expressing tubers did not lead to a decrease in total PPi content. However, the expression of the heterologous pyrophosphatase in the cytosol of cytosolic invertase-expressing tubers led to strong metabolic changes. These results are discussed both with respect to our previous hypotheses and to current models of the compartmentation of potato tuber metabolism.

  11. Probing riboswitch-ligand interactions using thiamine pyrophosphate analogues.

    PubMed

    Chen, Liuhong; Cressina, Elena; Dixon, Neil; Erixon, Karl; Agyei-Owusu, Kwasi; Micklefield, Jason; Smith, Alison G; Abell, Chris; Leeper, Finian J

    2012-08-14

    The Escherichia coli thiM riboswitch forms specific contacts with its natural ligand, thiamine pyrophosphate (TPP or thiamine diphosphate), allowing it to generate not only nanomolar binding affinity, but also a high degree of discrimination against similar small molecules. A range of synthetic TPP analogues have been used to probe each of the riboswitch-ligand interactions. The results show that the pyrimidine-sensing helix of thiM is exquisitely tuned to select for TPP by recognising the H-bonding donor and acceptors around its aminopyrimidine ring and also by forming π-stacking interactions that may be sensitive to the electronics of the ring. The central thiazolium ring of TPP appears to be more important for ligand recognition than previously thought. It may contribute to binding via long-range electrostatic interactions and/or by exerting an electron withdrawing effect on the pyrimidine ring, allowing its presence to be sensed indirectly and thereby allowing discrimination between thiamine (and its phosphate esters) and other aminopyrimidines found in vivo. The pyrophosphate moiety is essential for submicromolar binding affinity, but unexpectedly, it does not appear to be strictly necessary for modulation of gene expression.

  12. Micro-RNA quantification using DNA polymerase and pyrophosphate quantification.

    PubMed

    Yu, Hsiang-Ping; Hsiao, Yi-Ling; Pan, Hung-Yin; Huang, Chih-Hung; Hou, Shao-Yi

    2011-12-15

    A rapid quantification method for micro-RNA based on DNA polymerase activity and pyrophosphate quantification has been developed. The tested micro-RNA serves as the primer, unlike the DNA primer in all DNA sequencing methods, and the DNA probe serves as the template for DNA replication. After the DNA synthesis, the pyrophosphate detection and quantification indicate the existence and quantity of the tested miRNA. Five femtomoles of the synthetic RNA could be detected. In 20-100 μg RNA samples purified from SiHa cells, the measurement was done using the proposed assay in which hsa-miR-16 and hsa-miR-21 are 0.34 fmol/μg RNA and 0.71 fmol/μg RNA, respectively. This simple and inexpensive assay takes less than 5 min after total RNA purification and preparation. The quantification is not affected by the pre-miRNA which cannot serve as the primer for the DNA synthesis in this assay. This assay is general for the detection of the target RNA or DNA with a known matched DNA template probe, which could be widely used for detection of small RNA, messenger RNA, RNA viruses, and DNA. Therefore, the method could be widely used in RNA and DNA assays.

  13. A study about some phosphate derivatives as inhibitors of calcium oxalate crystal growth

    NASA Astrophysics Data System (ADS)

    Grases, F.; March, P.

    1989-08-01

    The kinetic of crystal growth of calcium oxalate monohydrate seed crystals were investigated potentiometrically in the presence of several phosphate derivatives, D-fructose-1,6-diphosphate, pyrophosphate, methylene diphosphonate and phytate, and it was found that in some cases they strongly inhibited crystal growth. The inhibitory action of the different substances assayed was comparatively evaluated.

  14. Evaluation of carbonate apatite blocks fabricated from dicalcium phosphate dihydrate blocks for reconstruction of rabbit femoral and tibial defects.

    PubMed

    Kanazawa, Masayuki; Tsuru, Kanji; Fukuda, Naoyuki; Sakemi, Yuta; Nakashima, Yasuharu; Ishikawa, Kunio

    2017-06-01

    This study aimed to evaluate in vivo behavior of a carbonate apatite (CO3Ap) block fabricated by compositional transformation via a dissolution-precipitation reaction using a calcium hydrogen phosphate dihydrate [DCPD: CaHPO4·2H2O] block as a precursor. These blocks were used to reconstruct defects in the femur and tibia of rabbits, using sintered dense hydroxyapatite (HAp) blocks as the control. Both the CO3Ap and HAp blocks showed excellent tissue response and good osteoconductivity. HAp block maintained its structure even after 24 weeks of implantation, so no bone replacement of the implant was observed throughout the post-implantation period in either femoral or tibial bone defects. In contrast, CO3Ap was resorbed with increasing time after implantation and replaced with new bone. The CO3Ap block was resorbed approximately twice as fast at the metaphysis of the proximal tibia than at the epiphysis of the distal femur. The CO3Ap block was resorbed at an approximately linear change over time, with complete resorption was estimated by extrapolation of data at approximately 1-1.5 years. Hence, the CO3Ap block fabricated in this study has potential value as an ideal artificial bone substitute because of its resorption and subsequent replacement by bone.

  15. Crystallization of dicalcium phosphate dihydrate with presence of glutamic acid and arginine at 37 °C.

    PubMed

    Li, Chengfeng; Ge, Xiaolu; Li, Guochang; Bai, Jiahai; Ding, Rui

    2014-08-01

    The formations of non-metabolic stones, bones and teeth were seriously related to the morphology, size and surface reactivity of dicalcium phosphate dihydrate (DCPD). Herein, a facile biomimetic mineralization method with presence of glutamic acid and arginine was employed to fabricate DCPD with well-defined morphology and adjustable crystallite size. In reaction solution containing more arginine, crystallization of DCPD occurred with faster rate of nucleation and higher density of stacked layers due to the generation of more OH(-) ions after hydrolysis of arginine at 37 °C. With addition of fluorescein or acetone, the consumption of OH(-) ions or desolvation reaction of Ca(2+) ions was modulated, which resulted in the fabrication of DCPD with adjustable crystallite sizes and densities of stacked layers. In comparison with fluorescein-loading DCPD, dicalcium phosphate anhydrate was prepared with enhanced photoluminescence properties due to the reduction of self-quenching effect and regular arrangement of encapsulated fluorescein molecules. With addition of more acetone, DCPD was prepared with smaller crystallite size via antisolvent crystallization. The simulated process with addition of amino acids under 37 °C would shed light on the dynamic process of biomineralization for calcium phosphate compounds.

  16. Enhanced triterpene accumulation in Panax ginseng hairy roots overexpressing mevalonate-5-pyrophosphate decarboxylase and farnesyl pyrophosphate synthase.

    PubMed

    Kim, Yong-Kyoung; Kim, Yeon Bok; Uddin, Md Romij; Lee, Sanghyun; Kim, Soo-Un; Park, Sang Un

    2014-10-17

    To elucidate the function of mevalonate-5-pyrophosphate decarboxylase (MVD) and farnesyl pyrophosphate synthase (FPS) in triterpene biosynthesis, the genes governing the expression of these enzymes were transformed into Panax ginseng hairy roots. All the transgenic lines showed higher expression levels of PgMVD and PgFPS than that by the wild-type control. Among the hairy root lines transformed with PgMVD, M18 showed the highest level of transcription compared to the control (14.5-fold higher). Transcriptions of F11 and F20 transformed with PgFPS showed 11.1-fold higher level compared with control. In triterpene analysis, M25 of PgMVD produced 4.4-fold higher stigmasterol content (138.95 μg/100 mg, dry weight [DW]) than that by the control; F17 of PgFPS showed the highest total ginsenoside (36.42 mg/g DW) content, which was 2.4-fold higher compared with control. Our results indicate that metabolic engineering in P. ginseng was successfully achieved through Agrobacterium rhizogenes-mediated transformation and that the accumulation of phytosterols and ginsenosides was enhanced by introducing the PgMVD and PgFPS genes into the hairy roots of the plant. Our results suggest that PgMVD and PgFPS play an important role in the triterpene biosynthesis of P. ginseng.

  17. Investigating the pharmacodynamic and magnetic properties of pyrophosphate-bridged coordination complexes

    NASA Astrophysics Data System (ADS)

    Ikotun, Oluwatayo (Tayo) F.

    The multidentate nature of pyrophosphate makes it an attractive ligand for complexation of metal cations. The participation of pyrophosphate in a variety of biological pathways and its metal catalyzed hydrolysis has driven our investigation into its coordination chemistry. We have successfully synthesized a library of binuclear pyrophosphate bridge coordination complexes. The problem of pyrophosphate hydrolysis to phosphate in the presence of divalent metal ions was overcome by incorporating capping ligands such as 1,10-phenanthroline and 2,2'-bipyridine prior to the addition of the pyrophosphate. The magnetic properties of these complexes was investigated and magneto-structural analysis was conducted. The biological abundance of pyrophosphate and the success of metal based drugs such as cisplatin, prompted our investigation of the cytotoxic properties of M(II) pyrophosphate dimeric complexes (where M(II) is CoII, CuII, and NiII) in adriamycin resistant human ovarian cancer cells. Thess compounds were found to exhibit toxicity in the nanomolar to picomolar range. We conducted in vitro stability studies and the mechanism of cytoxicity was elucidated by performing DNA mobility and binding assays, enzyme inhibition assays, and in vitro oxidative stress studies.

  18. Suicidal dephosphorylation of thiamine pyrophosphate coupled with pyruvate dehydrogenase complex.

    PubMed

    Strumilo, Slawomir; Dobrzyn, Pawel; Czerniecki, Jan; Tylicki, Adam

    2004-12-01

    Earlier it was noted that purified pyruvate dehydrogenase complex (PDC) produced by "Sigma" usually contains almost saturating amounts of thiamine pyrophosphate (ThPP). In this communication we present the observation that the endogenous ThPP coupled to PDC is dephosphorylated while staying at -10 degrees C, because in the enzyme preparation thiamine monophosphate and un-phosphorylated thiamine appear (HPLC determination). Under the same conditions exogenous ThPP is not dephosphorylated despite contact with the PDC preparation. This may suggest that interactions of some active groups of the enzyme with molecules of endogenous ThPP leads to break-up of the phosphoesters bonds, and destruction of the coenzyme. Decrease of PDC activity during storage is not in proportion with the degree of ThPP dephosphorylation. However the observed instability of PDC activity may be a consequence of the spontaneous process of its coenzyme autodestruction.

  19. Identification of novel ligands for thiamine pyrophosphate (TPP) riboswitches.

    PubMed

    Cressina, Elena; Chen, Liuhong; Moulin, Michael; Leeper, Finian J; Abell, Chris; Smith, Alison G

    2011-04-01

    Riboswitches are regions of mRNA to which a metabolite binds in the absence of proteins, resoulting in alteration of transcription, translation or splicing. The most widespread forms of riboswitches are those responsive to TPP (thiamine pyrophosphate) the active form of vitamin B1, thiamine. TPP-riboswitches have been found in all bacterial genomes examined, and are the only ones found in eukaryotes. In each case, the riboswitch appears to regulate the expression of a gene involved in synthesis or uptake of the vitamin. Riboswitches offer an attractive target for chemical intervention, and identification of novel ligands would allow a detailed study on structure-activity relationships, as well as potential leads for the development of antimicrobial compounds. To this end, we have developed a medium-throughput methodology for screening libraries of small molecules using biophysical methods.

  20. Effects of farnesyl pyrophosphate accumulation on calvarial osteoblast differentiation.

    PubMed

    Weivoda, Megan M; Hohl, Raymond J

    2011-08-01

    Statins, drugs commonly used to lower serum cholesterol, have been shown to stimulate osteoblast differentiation and bone formation. Statins inhibit 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A reductase (HMGCR), the first step of the isoprenoid biosynthetic pathway, leading to the depletion of the isoprenoids farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP). The effects of statins on bone have previously been attributed to the depletion of GGPP, because the addition of exogenous GGPP prevented statin-stimulated osteoblast differentiation in vitro. However, in a recent report, we demonstrated that the specific depletion of GGPP did not stimulate but, in fact, inhibited osteoblast differentiation. This led us to hypothesize that isoprenoids upstream of GGPP play a role in the regulation of osteoblast differentiation. We demonstrate here that the expression of HMGCR and FPP synthase decreased during primary calvarial osteoblast differentiation, correlating with decreased FPP and GGPP levels during differentiation. Zaragozic acid (ZGA) inhibits the isoprenoid biosynthetic pathway enzyme squalene synthase, leading to an accumulation of the squalene synthase substrate FPP. ZGA treatment of calvarial osteoblasts led to a significant increase in intracellular FPP and resulted in inhibition of osteoblast differentiation as measured by osteoblastic gene expression, alkaline phosphatase activity, and matrix mineralization. Simultaneous HMGCR inhibition prevented the accumulation of FPP and restored osteoblast differentiation. In contrast, specifically inhibiting GGPPS to lower the ZGA-induced increase in GGPP did not restore osteoblast differentiation. The specificity of HMGCR inhibition to restore osteoblast differentiation of ZGA-treated cultures through the reduction in isoprenoid accumulation was confirmed with the addition of exogenous mevalonate. Similar to ZGA treatment, exogenous FPP inhibited the mineralization of primary calvarial osteoblasts

  1. The Role of Inorganic Pyrophosphate in Aortic Valve Calcification

    PubMed Central

    Rathan, Swetha; Yoganathan, Ajit P.; O’Neill, W. Charles

    2017-01-01

    Background and aim of the study Aortic valve (AV) calcification is a major cause of morbidity and mortality, yet the molecular mechanisms involved are poorly understood. Hence, an ex vivo model of calcification in intact AVs was developed in order to test the role of orthophosphate and pyrophosphate (PPi), both of which factors are known to influence vascular calcification. Methods Porcine AV leaflets were cultured in serum-free medium under static conditions for eight days, over which time leaflet architecture and viability were preserved. Calcification was measured as the incorporation of 45Ca, with confirmation by Alizarin Red staining. Results Calcification required both a high phosphate concentration (3.8 mM) and removal of PPi with alkaline phosphatase or inorganic pyrophosphatase. Calcification occurred predominantly on the fibrosa and was arrested by the bisphosphonate etidronate, a non-hydrolyzable analog of PPi. Leaflets released PPi into the medium, and this was enhanced by MLS38949, a specific inhibitor of tissue non-specific alkaline phosphatase (TNAP). Furthermore, leaflets synthesized PPi from extracellular ATP, which was reduced by β, γ-methylene-ATP, an inhibitor of ectonucleotide pyrophosphorylase phosphodiesterase (NPP1). Conclusion The ex vivo AV calcification model developed in the present study showed that extracellular PPi, produced by valvular tissue, is a potent inhibitor of valvular calcification. In addition to synthesis, hydrolysis by TNAP also controls PPi levels and calcification. The results suggest that a decreased synthesis or increased hydrolysis of pyrophosphate may contribute to valvular calcification, and that bisphosphonates or inhibitors of TNAP are potential preventive strategies of the process. TNAP are potential preventive strategies. PMID:25803964

  2. Effect of pyrophosphate ions on the conversion of calcium–lithium–borate glass to hydroxyapatite in aqueous phosphate solution

    PubMed Central

    Fu, Hailuo; Day, Delbert E.; Huang, Wenhai

    2010-01-01

    The conversion of glass to a hydroxyapatite (HA) material in an aqueous phosphate solution is used as an indication of the bioactive potential of the glass, as well as a low temperature route for preparing biologically useful materials. In this work, the effect of varying concentrations of pyrophosphate ions in the phosphate solution on the conversion of a calcium–lithium–borate glass to HA was investigated. Particles of the glass (150–355 µm) were immersed for up to 28 days in 0.25 M K2HPO4 solution containing 0–0.1 M K4P2O7. The kinetics of degradation of the glass particles and their conversion to HA were monitored by measuring the weight loss of the particles and the ionic concentration of the solution. The structure and composition of the conversion products were analyzed using X-ray diffraction, scanning electron microscopy, and Fourier transform infrared spectroscopy. For K4P2O7 concentrations of up to 0.01 M, the glass particles converted to HA, but the time for complete conversion increased from 2 days (no K4P2O7) to 10 days (0.01 M K4P2O7). When the K4P2O7 concentration was increased to 0.1 M, the product consisted of an amorphous calcium phosphate material, which eventually crystallized to a pyrophosphate product (predominantly K2CaP2O7 and Ca2P2O7). The consequences of the results for the formation of HA materials and devices by the glass conversion route are discussed. PMID:20680413

  3. A review on the chemical synthesis of pyrophosphate bonds in bioactive nucleoside diphosphate analogs.

    PubMed

    Xu, Zhihong

    2015-09-15

    Currently, there is an ongoing interest in the synthesis of nucleoside diphosphate analogs as important regulators in catabolism/anabolism, and their potential applications as mechanistic probes and chemical tools for bioassays. However, the pyrophosphate bond formation step remains as the bottleneck. In this Digest, the chemical synthesis of the pyrophosphate bonds of representative bioactive nucleoside diphosphate analogs, i.e. phosphorus-modified analogs, nucleoside cyclic diphosphates, and nucleoside diphosphate conjugates, will be described.

  4. Inositol pyrophosphates modulate cell cycle independently of alteration in telomere length.

    PubMed

    Banfic, Hrvoje; Crljen, Vladiana; Lukinovic-Skudar, Vesna; Dembitz, Vilma; Lalic, Hrvoje; Bedalov, Antonio; Visnjic, Dora

    2016-01-01

    Synthesis of inositol pyrophosphates through activation of Kcs1 plays an important role in the signalling response required for cell cycle progression after mating pheromone arrest. Overexpression of Kcs1 doubled the level of inositol pyrophosphates when compared to wild type cells and 30 min following the release from α-factor block further increase in inositol pyrophosphates was observed, which resulted that cells overexpressing Kcs1 reached G2/M phase earlier than wild type cells. Similar effect was observed in ipk1Δ cells, which are unable to synthesize IP6-derived inositol pyrophosphates (IP7 and IP8) but will synthesize IP5-derived inositol pyrophosphates (PP-IP4 and (PP)2-IP3). Although ipk1Δ cells have shorter telomeres than wild type cells, overexpression of Kcs1 in both strains have similar effect on cell cycle progression. As it is known that PP-IP4 regulates telomere length through Tel1, inositol polyphosphates, cell cycle and telomere length were determined in tel1Δ cells. The release of the cells from α-factor block and overexpression of Kcs1 in tel1Δ cells produced similar effects on inositol pyrophosphates level and cell cycle progression when compared to wild type cells, although tel1Δ cells possesses shorter telomeres than wild type cells. It can be concluded that telomere length does not affect cell cycle progression, since cells with short telomeres (ipk1Δ and tel1Δ) progress through cell cycle in a similar manner as wild type cells and that overexpression of Kcs1 in cells with either short or normal telomeres will increase S phase progression without affecting telomere length. Furthermore, IP5-derived inositol pyrophosphates can compensate for the loss of IP6-derived inositol pyrophosphates, in modulating S phase progression of the cell cycle.

  5. Calcium Carbonate

    MedlinePlus

    Calcium carbonate is a dietary supplement used when the amount of calcium taken in the diet is not ... for healthy bones, muscles, nervous system, and heart. Calcium carbonate also is used as an antacid to relieve ...

  6. Development of an enhanced anticaries efficacy dual component dentifrice containing sodium fluoride and dicalcium phosphate dihydrate.

    PubMed

    Sullivan, R J; Masters, J; Cantore, R; Roberson, A; Petrou, I; Stranick, M; Goldman, H; Guggenheim, B; Gaffar, A

    2001-05-01

    A dual-chamber dentifrice, which contains sodium fluoride (NaF) in one component and dicalcium phosphate dihydrate (dical) in the other, has been developed. The dentifrice is packaged in a dual-chamber tube and is formulated to deliver 1100 ppm F. A series of studies consisting of in vitro fluoride uptake, in vivo calcium labeling, intraoral remineralization-demineralization, and animal caries studies were performed to support the improved anticaries efficacy of this product in comparison to a sodium fluoride/silica dentifrice (NaF/silica). An in vitro fluoride uptake study comparing 1100 ppm F NaF/dical dentifrice to 1100 ppm F NaF/silica showed that NaF/dical delivered significantly more fluoride than NaF/silica, 3.72 +/- 0.36 micrograms/cm2 versus 2.41 +/- 0.10 micrograms/cm2. A 6-day in vivo brushing study with a 44Ca labeled NaF/dical dentifrice showed that calcium from dical penetrated demineralized enamel and was present in plaque up to 18 hrs since the last brushing. An intra-oral remineralization-demineralization study was performed to evaluate NaF/dical's ability to promote remineralization in comparison to three silica-based dentifrices containing 0, 250, and 1100 ppm F as NaF. The percent mineral changes after treatment were +20.44 +/- 17.14 for NaF/dical, +9.27 +/- 19.53 for 1100 ppm NaF/silica, -1.43 +/- 20.57 for 250 ppm NaF/silica, and -12.36 +/- 32.76 for 0 ppm F/silica. A statistical analysis showed that the dual-chamber NaF/dical dentifrice was significantly more effective than the 1100 ppm NaF/silica dentifrice at promoting remineralization. A rat caries study was performed to evaluate NaF/dical ability to prevent caries in comparison to 1100 ppm F NaF/silica, 250 ppm F NaF/silica, silica, and dical dentifrices. The mean smooth surface caries scores were 1.6 +/- 2.8 for NaF/dical, 5.5 +/- 6.2 for 1100 ppm F NaF/silica, 10.6 +/- 6.2 for 250 ppm F NaF/silica, 13.7 +/- 4.7 for 0 ppm F/silica, and 9.5 +/- 7.8 0 ppm F/dical. A statistical analysis

  7. IdsA is the major geranylgeranyl pyrophosphate synthase involved in carotenogenesis in Corynebacterium glutamicum.

    PubMed

    Heider, Sabine A E; Peters-Wendisch, Petra; Beekwilder, Jules; Wendisch, Volker F

    2014-11-01

    Corynebacterium glutamicum, a yellow-pigmented soil bacterium that synthesizes the rare cyclic C50 carotenoid decaprenoxanthin and its glucosides, has been engineered for the production of various carotenoids. CrtE was assumed to be the major geranylgeranyl pyrophosphate (GGPP) synthase in carotenogenesis; however, deletion of crtE did not abrogate carotenoid synthesis. In silico analysis of the repertoire of prenyltransferases encoded by the C. glutamicum genome revealed two candidate GGPPS genes (idsA and ispB). The absence of pigmentation of an idsA deletion mutant and complementation experiments with a double deletion mutant lacking both idsA and crtE showed that IdsA is the major GGPPS of C. glutamicum and that crtE overexpression compensated for the lack of IdsA, whereas plasmid-borne overexpression of ispB did not. Purified His-tagged CrtE was active as a homodimer, whereas the active form of IdsA was homotetrameric. Both enzymes catalyzed prenyl transfer with isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate, geranyl pyrophosphate and farnesylphosphate (FPP) as substrates. IdsA showed the highest catalytic efficiency with dimethylallyl pyrophosphate and IPP, whereas the catalytic efficiency of CrtE was highest with geranyl pyrophosphate and IPP. Finally, application of prenyltransferase overexpression revealed that combined overexpression of idsA and the IPP isomerase gene idi in the absence of crtE led to the highest decaprenoxanthin titer reported to date.

  8. Protein-Mediated Precipitation of Calcium Carbonate

    PubMed Central

    Polowczyk, Izabela; Bastrzyk, Anna; Fiedot, Marta

    2016-01-01

    Calcium carbonate is an important component in exoskeletons of many organisms. The synthesis of calcium carbonate was performed by mixing dimethyl carbonate and an aqueous solution of calcium chloride dihydrate. The precipitation product was characterized by means of scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) measurements. In addition, the turbidity of the reaction solution was acquired to monitor the kinetics of the calcium carbonate structure’s growth in the investigated system. In this study, samples of CaCO3 particles obtained with individual proteins, such as ovalbumin, lysozyme, and a mixture of the proteins, were characterized and compared with a control sample, i.e., synthesized without proteins. The obtained data indicated that the addition of ovalbumin to the reaction changed the morphology of crystals from rhombohedral to ‘stack-like’ structures. Lysozyme, however, did not affect the morphology of calcium carbonate, yet the presence of the protein mixture led to the creation of more complex composites in which the calcium carbonate crystals were constructed in protein matrices formed by the ovalbumin-lysozyme interaction. It was also observed that in the protein mixture, ovalbumin has a major influence on the CaCO3 formation through a strong interaction with calcium ions, which leads to the coalescence and creation of a steric barrier reducing particle growth. The authors proposed a mechanism of calcium carbonate grain growth in the presence of both proteins, taking into account the interaction of calcium ions with the protein. PMID:28774065

  9. Solubility of Calcium Phosphate in Concentrated Dairy Effluent Brines.

    PubMed

    Kezia, K; Lee, J; Zisu, B; Chen, G Q; Gras, S L; Kentish, S E

    2017-05-24

    The solubility of calcium phosphate in concentrated dairy brine streams is important in understanding mineral scaling on equipment, such as membrane modules, evaporators, and heat exchangers, and in brine pond operation. In this study, the solubility of calcium phosphate has been assessed in the presence of up to 300 g/L sodium chloride as well as lactose, organic acids, and anions at 10, 30, and 50 °C. As a neutral molecule, lactose has a marginal but still detectable effect upon calcium solubility. However, additions of sodium chloride up to 100 g/L result in a much greater increase in calcium solubility. Beyond this point, the concentrations of ions in the solution decrease significantly. These changes in calcium solubility can readily be explained through changes in the activity coefficients. There is little difference in calcium phosphate speciation between 10 and 30 °C. However, at 50 °C, the ratio of calcium to phosphate in the solution is lower than at the other temperatures and varies less with ionic strength. While the addition of sodium lactate has less effect upon calcium solubility than sodium citrate, it still has a greater effect than sodium chloride at an equivalent ionic strength. Conversely, when these organic anions are present in the solution in the acid form, the effect of pH dominates and results in much higher solubility and a calcium/phosphate ratio close to one, indicative of dicalcium phosphate dihydrate as the dominant solid phase.

  10. Crystal arthritides - gout and calcium pyrophosphate arthritis : Part 3: Treatment.

    PubMed

    Schlee, S; Bollheimer, L C; Bertsch, T; Sieber, C C; Härle, P

    2017-02-28

    The treatment of gout is based on several principles. Symptom control and termination of the inflammatory process are important early goals, whereas the urate level should be lowered in the long term to prevent further gout attacks and complications. The non-pharmacological approach is based on individually informing the patient on dietary measures and changes of life style. Besides physical measures, such as cold applications on the affected joint, various medications are available for treatment of an acute gout attack. The choice of drug depends on the individual risk profile. If non-steroidal anti-inflammatory drugs (NSAID) and coxibs are chosen it should be taken into account that the use is restricted in patients with renal insufficiency. Moreover, these drugs may have gastrointestinal side effects and are associated with increased cardiovascular morbidity and mortality. Colchicine has gastrointestinal side effects at high dosages but can also be used for differential diagnostics if there is a quick response to treatment. Steroids are an effective alternative and can be given orally or parenterally in patients with dysphagia. Moreover, steroids can be used in cases of renal insufficiency. After symptoms of the acute attack have subsided, urate lowering therapy should be initiated to prevent further attacks. Low-dose urate lowering therapy can be started during an acute gout attack when acute therapy is initiated. Allopurinol is still the medication of choice but its use is restricted in patients with renal insufficiency. A rare but serious side effect is allopurinol hypersensitivity syndrome. Febuxostat can be an alternative in patients who do not tolerate allopurinol. In February 2016, lesinurad, an URAT-1 and OAT-4 inhibitor, was approved in combination with allopurinol or febuxostat. Data on the effectiveness and safety of synthetic uricases and biologicals are still sparse for elderly patients. These substances are reserved for severe cases of gout.

  11. Crystal structures of the thi-box riboswitch bound to thiamine pyrophosphate analogs reveal adaptive RNA-small molecule recognition.

    PubMed

    Edwards, Thomas E; Ferré-D'Amaré, Adrian R

    2006-09-01

    Riboswitches are noncoding mRNA elements that bind small-molecule metabolites with high affinity and specificity, and they regulate the expression of associated genes. The thi-box riboswitch can exhibit a 1000-fold higher affinity for thiamine pyrophosphate over closely related noncognate compounds such as thiamine monophosphate. To understand the chemical basis of thi-box pyrophosphate specificity, we have determined crystal structures of an E. coli thi-box bound to thiamine pyrophosphate, thiamine monophosphate, and the structural analogs benfotiamine and pyrithiamine. When bound to monophosphorylated compounds, the RNA elements that recognize the thiamine and phosphate moieties of the ligand move closer together. This allows the riboswitch to recognize the monophosphate in a manner similar to how it recognizes the beta-phosphate of thiamine pyrophosphate. In the pyrithiamine complex, the pyrophosphate binding site is largely unstructured. These results show how the riboswitch can bind to various metabolites, and why the thi-box preferentially binds thiamine pyrophosphate.

  12. Biosynthesis of monoterpenes: Stereochemistry of the coupled isomerization and cyclization of geranyl pyrophosphate to camphane and isocamphane monoterpenes

    SciTech Connect

    Croteau, R.; Gershenzon, J.; Wheeler, C.J.; Satterwhite, D.M. )

    1990-03-01

    The conversion of geranyl pyrophosphate to (+)-bornyl pyrophosphate and (+)-camphene is considered to proceed by the initial isomerization of the substrate to (-)-(3R)-linalyl pyrophosphate and the subsequent cyclization of this bound intermediate. In the case of (-)-bornyl pyrophosphate and (-)-camphene, isomerization of the substrate to the (+)-(3S)-linalyl intermediate precedes cyclization. The geranyl and linalyl precursors were shown to be mutually competitive substrates (inhibitors) of the relevant cyclization enzymes isolated from Salvia officinalis (sage) and Tanacetum vulgare (tansy) by the mixed substrate analysis method, demonstrating that isomerization and cyclization take place at the same active site. Incubation of partially purified enzyme preparations with (3R)-(1Z-3H)linalyl pyrophosphate plus (1-14C)geranyl pyrophosphate gave rise to double-labeled (+)-bornyl pyrophosphate and (+)-camphene, whereas incubation of enzyme preparations catalyzing the antipodal cyclizations with (3S)-(1Z-3H)-linalyl pyrophosphate plus (1-14C)geranyl pyrophosphate yielded double-labeled (-)-bornyl pyrophosphate and (-)-camphene. Each product was then transformed to the corresponding (+)- or (-)-camphor without change in the 3H:14C isotope ratio, and the location of the tritium label was deduced in each case by stereoselective, base-catalyzed exchange of the exo-alpha-hydrogen of the derived ketone. The finding that the 1Z-3H of the linalyl precursor was positioned at the endo-alpha-hydrogen of the corresponding camphor in all cases, coupled to the previously demonstrated retention of configuration at C1 of the geranyl substrate in these transformations, confirmed the syn-isomerization of geranyl pyrophosphate to linalyl pyrophosphate and the cyclization of the latter via the anti,endo- conformer.

  13. Calpain-1 Mediated Disorder of Pyrophosphate Metabolism Contributes to Vascular Calcification Induced by oxLDL

    PubMed Central

    Lu, Meili; Zhang, Xiaowen; Dai, Chunmei; Mei, Meng; Zhang, Suping; Wang, Hongxin; Song, Qing

    2015-01-01

    We previously reported that oxidized low density lipoprotein (oxLDL) accelerated the calcification in aorta of rats and rat vascular smooth muscle cells (RVSMCs). However, the molecular mechanism underlying the acceleration remains poorly understood. The present study aimed to investigate the role of calpain-1, Ca2+-sensitive intracellular cysteine proteases, in the vascular calcification of rats treated with both high dose of vitamin D2 and high cholesterol diet. The results showed that calpain activity significantly increased in calcified aortic tissue of rats and RVSMCs treated with oxLDL. Specific calpain inhibitor I (CAI, 0.5mg/kg, intraperitoneal) inhibited the vascular calcification in rats with hypercholesterolemia accompanied by the increase in the level of extracellular inorganic pyrophosphate (PPi), the endogenous inhibitor of vascular calcification. In addition, CAI increased the content of adenosine triphosphate (ATP), decreased the activity, mRNA and protein expression of alkaline phosphatase (ALP) and reduced the production of superoxide anion in calcified aortic tissue. CAI also increased the activity of ATP synthase as well as protein expression of ATP5D, δ subunit of ATP synthase. In the in vitro study, suppression of calpain-1 using siRNA assay inhibited the calcium deposition, increased the levels of PPi and ATP, improved the activity of ATP synthase as well as protein expression of ATP5D in RVSMCs treated with oxLDL. Calpain-1 suppression also decreased the activity, mRNA and protein expression of ALP and reduced the mitochondrial ROS (Mito-ROS) production in RVSMCs. However, mito-TEMPO, the mitochondria-targeted ROS scavenger, reduced the calcium deposition, increased the PPi in culture medium, decreased the activity, mRNA and protein expression of ALP in RVSMCs treated with oxLDL. Taken together, the results suggested that calpain-1 activation plays critical role in vascular calcification caused by oxLDL, which might be mediated by PPi

  14. Destructive wrist arthropathy of pseudogout

    SciTech Connect

    Smathers, R.L.; Keats, T.E.; Stelling, C.B.

    1982-01-01

    The wrist is a frequent site of arthropathy in the pseudogout syndrome (calcium pyrophosphate dihydrate crystal deposition disease). Three cases of severe wrist arthropathy are presented to emphasize the destructive changes which may occur, including necrosis or collapse of the lunate and navicular bones. The roentgenographic findings characteristic of pseudogout in the wrist are discussed.

  15. The crystallization of hydroxyapatite and dicalcium phosphate dihydrate; representation of growth curves

    NASA Astrophysics Data System (ADS)

    Hohl, H.; Koutsoukos, P. G.; Nancollas, G. H.

    1982-04-01

    The kinetics of growth of dicalcium phosphate dihydrate seed crystals has been investigated by a method in which the activities of the lattice ions were maintained constant during the reaction. The constant composition procedure has also been used to study the crystallization of hydroxyapatite at very low supersaturation. Changes in specific area during the resulting appreciable extents of growth have been used to predict the developing morphologies of the crystals. For dicalcium phosphate dihydrate and hydroxyapatite, the predominant crystal growth takes place in two dimensions and one dimension, respectively. In both cases the growth rates are pH dependent. Dimensionless representation of the crystallization rates and driving forces enable comparisons to be made between data for electrolytes of different charge types.

  16. Crystal structure of strychninium chloride dihydrate: Hidden helix in the water/anion tape

    NASA Astrophysics Data System (ADS)

    Białońska, Agata; Ciunik, Zbigniew

    2005-11-01

    The crystal structure of strychninium chloride dihydrate SH +Cl -·2H 2O was determined. The structure is composed of strychnine herring-bone bilayer sheets with channels occupied by the hydrogen bonded anion/water tape between them. Considering different amount of water molecules in isomorphous crystal of SH +Cl -·2H 2O and previously described crystal of strychninium chloride sesquihydrate, we found that water molecules and chloride anions form a left-handed helix. Similarly, anions and water molecules in the crystal of strychnine bromide dihydrate form a left-handed helix related by the two-fold screw axis symmetry. Contrary, in the crystals of strychninium chloride sesquihydrate and SH +Cl -·2H 2O, the helices are related by only translation vector and are stabilized by one or two water bridges, respectively.

  17. Cellular Cations Control Conformational Switching of Inositol Pyrophosphate Analogues.

    PubMed

    Hager, Anastasia; Wu, Mingxuan; Wang, Huanchen; Brown, Nathaniel W; Shears, Stephen B; Veiga, Nicolás; Fiedler, Dorothea

    2016-08-22

    The inositol pyrophosphate messengers (PP-InsPs) are emerging as an important class of cellular regulators. These molecules have been linked to numerous biological processes, including insulin secretion and cancer cell migration, but how they trigger such a wide range of cellular responses has remained unanswered in many cases. Here, we show that the PP-InsPs exhibit complex speciation behaviour and propose that a unique conformational switching mechanism could contribute to their multifunctional effects. We synthesised non-hydrolysable bisphosphonate analogues and crystallised the analogues in complex with mammalian PPIP5K2 kinase. Subsequently, the bisphosphonate analogues were used to investigate the protonation sequence, metal-coordination properties, and conformation in solution. Remarkably, the presence of potassium and magnesium ions enabled the analogues to adopt two different conformations near physiological pH. Understanding how the intrinsic chemical properties of the PP-InsPs can contribute to their complex signalling outputs will be essential to elucidate their regulatory functions.

  18. Thiamine pyrophosphate biosynthesis and transport in the nematode Caenorhabditis elegans.

    PubMed

    de Jong, Liesbeth; Meng, Yan; Dent, Joseph; Hekimi, Siegfried

    2004-10-01

    Thiamine (vitamin B1) is required in the diet of animals, and thiamine deficiency leads to diseases such as beri-beri and the Wernicke-Korsakoff syndrome. Dietary thiamine (vitamin B1) consists mainly of thiamine pyrophosphate (TPP), which is transformed into thiamine by gastrointestinal phosphatases before absorption. It is believed that TPP itself cannot be transported across plasma membranes in significant amounts. We have identified a partial loss-of-function mutation in the Caenorhabditis elegans gene (tpk-1) that encodes thiamine pyrophosphokinase, which forms TPP from thiamine at the expense of ATP inside cells. The mutation slows physiological rhythms and the phenotype it produces can be rescued by TPP but not thiamine supplementation. tpk-1 functions cell nonautonomously, as the expression of wild-type tpk-1 in one tissue can rescue the function of other tissues that express only mutant tpk-1. These observations indicate that, in contrast to expectation from previous evidence, TPP can be transported across cell membranes. We also find that thiamine supplementation partially rescues the phenotype of partial loss-of-function mutants of the Na/K ATPase, providing genetic evidence that thiamine absorption, and/or redistribution from the absorbing cells, requires the full activity of this enzyme.

  19. Folding energy landscape of the thiamine pyrophosphate riboswitch aptamer.

    PubMed

    Anthony, Peter C; Perez, Christian F; García-García, Cuauhtémoc; Block, Steven M

    2012-01-31

    Riboswitches are motifs in the untranslated regions (UTRs) of RNA transcripts that sense metabolite levels and modulate the expression of the corresponding genes for metabolite import, export, synthesis, or degradation. All riboswitches contain an aptamer: an RNA structure that, upon binding ligand, folds to expose or sequester regulatory elements in the adjacent sequence through alternative nucleotide pairing. The coupling between ligand binding and aptamer folding is central to the regulatory mechanisms of thiamine pyrophosphate (TPP) riboswitches and has not been fully characterized. Here, we show that TPP aptamer folding can be decomposed into ligand-independent and -dependent steps that correspond to the formation of secondary and tertiary structures, respectively. We reconstructed the full energy landscape for folding of the wild-type (WT) aptamer and measured perturbations of this landscape arising from mutations or ligand binding. We show that TPP binding proceeds in two steps, from a weakly to a strongly bound state. Our data imply a hierarchical folding sequence, and provide a framework for understanding molecular mechanism throughout the TPP riboswitch family.

  20. A Multi-Enzyme Bioluminescent Time-Resolved Pyrophosphate Assay

    PubMed Central

    Sun, Ye; Jacobson, K. Bruce; Golovlev, Val

    2009-01-01

    We have developed a high-sensitivity assay for measurement of inorganic pyrophosphate (PPi) in adenosine 5′-triphosphate (ATP) contaminated samples. The assay is based on time-resolved measurements of the luminescence kinetics and implements multiple enzymes to convert PPi to ATP that is, in turn, utilized to produce light; and to hydrolyze PPi for measurement of the steady-state background luminescence. A theoretical model for describing luminescence kinetics and optimizing composition of the assay detection mixture is presented. We found the model is in excellent agreement with the experimental results. We have developed and evaluated two algorithms for PPi measurement from luminescence kinetics acquired from ATP-contaminated samples. The first algorithm is considered to be the method of choice for analysis of long, i.e., 3-5 min, kinetics. The activity of enzymes is controlled during the experiment; the sensitivity of PPi detection is about 7 pg/ml or 15 pM of PPi in ATP-contaminated samples. The second algorithm is designed for analysis of short, i.e., less than 1-min luminescence kinetics. It has about 20 pM PPi detection sensitivity and may be the better choice for assays in microplate format, where a short measurement time is required. The PPi assay is primarily developed for RNA expression analysis, but it also can be used in various applications, which require high sensitivity PPi detection in ATP-contaminated samples. PMID:17540325

  1. PMBP extraction and TPE separation in alkaline pyrophosphate solutions

    SciTech Connect

    Karalova, Z.K.; Bukina, T.I.; Lavrinovich, E.A.; Trofimov, T.I.; Kulyako, Yu.M.; Myasoedov, B.F.

    1988-11-01

    Measurements have been made on the solvent extraction of Am(III), Am(V), Cm(III), and Eu(III) in trace and macroscopic amounts from (NH/sub 4/)/sub 4/P/sub 2/O/sub 7/ (pH = 7.6) and Na/sub 4/P/sub 2/O/sub 7/ (pH = 10) solutions as coordination compounds with 1-phenyl-3-methyl-4-benzoylpyrazol-5-one (PMBP). A method has been devised for separating weighable amounts of americium and curium, which is based in trivalent curium being extracted quantitatively from 0.1 M sodium pyrophosphate (pH 10) by 0.1 M PMBP solution in chloroform, where high distribution coefficients apply, while americium is electrochemically oxidized in that medium to Am(VI) and on contact with the extraction agent is reduced to Am(V) and remains in the aqueous phase. The separation factor for the Cm(III)-Am(V) pair is about 10/sup 3/.

  2. Protein farnesyltransferase: kinetics of farnesyl pyrophosphate binding and product release.

    PubMed

    Furfine, E S; Leban, J J; Landavazo, A; Moomaw, J F; Casey, P J

    1995-05-23

    Protein farnesyltransferase (FTase) catalyzes the prenylation of Ras and several other key proteins involved in cell regulation. The mechanism of the FTase reaction was elucidated by pre-steady-state and steady-state kinetic analysis. FTase catalyzed the farnesylation of biotinylated peptide substrate (BiopepSH) by farnesyl pyrophosphate (FPP) to an S-farnesylated peptide (BiopepS-C15). The steady-state kinetic mechanism was ordered. FTase bound FPP in a two-step process with an effective dissociation rate constant of 0.013 s-1 and an overall Kd of 2.8 nM. BiopepSH reacted with FTase.FPP irreversibly, with a second-order rate constant of 2.2 x 10(5) M-1 s-1, to form FTase.BiopepS-C15. Because most of the FPP in FTase.FPP was trapped as FTase.BiopepS-C15 at high concentrations of BiopepSH, FPP dissociated slowly from the ternary complex relative to catalysis, so that the commitment to catalysis was high. The maximal rate constant for formation of FTase.BiopepS-C15 (enzyme-bound product) is much larger than kcat (0.06 s-1), indicating that product release is the rate-determining step in the reaction mechanism.

  3. Farnesyl pyrophosphate synthase modulators: a patent review (2006 - 2010)

    PubMed Central

    Sun, Shuting; McKenna, Charles E.

    2012-01-01

    Introduction Farnesyl pyrophosphosphate synthase (FPPS (also known as farnesyl diphosphate synthase, FDPS)) is one of the key enzymes involved in the mevalonate pathway and as such is widely expressed. FPPS modulators, specifically FPPS inhibitors, are useful in treating a number of diseases, including bone related disorders characterized by excessive bone resorption e.g. osteoporosis, cancer metathesis to bone and infectious diseases caused by certain parasites. Areas covered This review covers structures and applications of novel FPPS modulators described in the patent literature from 2006 to 2010. Patents disclosing new formulations and uses of existing FPPS inhibitors are also reviewed. Thirty-three patents retrieved from the USPTO, EP and WIPO databases are examined with the goal of defining current trends in drug discovery related to FPPS inhibition, and its therapeutic effects. Expert opinion Bisphosphonates continue to dominate in this area, although other types of modulator are making their appearance. Remarkable for their high bone mineral affinity, bisphosphonates are structural mimics of the dimethylallyl pyrophosphate (DMAPP) substrate of FPPS, and constitute the major type of FPPS inhibitor currently used in the clinic for treatment of bone-related diseases. Lipophilic bisphosphonates and new classes of non-bisphosphonate FPPS inhibitors (salicylic acid and quinoline derivatives) have been introduced as possible alternatives for treatment of soft tissue diseases, such as some cancers. Novel formulations, fluorescent diagnostic probes and new therapeutic applications of existing FPPS inhibitors are also areas of significant patent activity, demonstrating growing recognition of the versatility and underdeveloped potential of these drugs. PMID:21702715

  4. Links between hydrothermal environments, pyrophosphate, na(+), and early evolution.

    PubMed

    Holm, Nils G; Baltscheffsky, Herrick

    2011-10-01

    The discovery that photosynthetic bacterial membrane-bound inorganic pyrophosphatase (PPase) catalyzed light-induced phosphorylation of orthophosphate (Pi) to pyrophosphate (PPi) and the capability of PPi to drive energy requiring dark reactions supported PPi as a possible early alternative to ATP. Like the proton-pumping ATPase, the corresponding membrane-bound PPase also is a H(+)-pump, and like the Na(+)-pumping ATPase, it can be a Na(+)-pump, both in archaeal and bacterial membranes. We suggest that PPi and Na(+) transport preceded ATP and H(+) transport in association with geochemistry of the Earth at the time of the origin and early evolution of life. Life may have started in connection with early plate tectonic processes coupled to alkaline hydrothermal activity. A hydrothermal environment in which Na(+) is abundant exists in sediment-starved subduction zones, like the Mariana forearc in the W Pacific Ocean. It is considered to mimic the Archean Earth. The forearc pore fluids have a pH up to 12.6, a Na(+)-concentration of 0.7 mol/kg seawater. PPi could have been formed during early subduction of oceanic lithosphere by dehydration of protonated orthophosphates. A key to PPi formation in these geological environments is a low local activity of water.

  5. Hierarchy of Pyrophosphate-Functionalized Uranyl Peroxide Nanocluster Synthesis.

    PubMed

    Dembowski, Mateusz; Colla, Christopher A; Hickam, Sarah; Oliveri, Anna F; Szymanowski, Jennifer E S; Oliver, Allen G; Casey, William H; Burns, Peter C

    2017-04-10

    Herein, we report a new salt of a pyrophosphate-functionalized uranyl peroxide nanocluster {U24Pp12} (1) exhibiting Oh molecular symmetry both in the solid and solution. Study of the system yielding 1 across a wide range of pH by single-crystal X-ray diffraction, small-angle X-ray scattering, and a combination of traditional (31)P and diffusion-ordered spectroscopy (DOSY) NMR affords unprecedented insight into the amphoteric chemistry of this uranyl peroxide system. Key results include formation of a rare binary {U24}·{U24Pp12} (3) system observed under alkaline conditions, and evidence of acid-promoted decomposition of {U24Pp12} (1) followed by spatial rearrangement and condensation of {U4} building blocks into the {U32Pp16} (2) cluster. Furthermore, (31)P DOSY NMR measurements performed on saturated solutions containing crystalline {U32Pp16} show only trace amounts (∼2% relative abundance) of the intact form of this cluster, suggesting a complex interconversion of {U24Pp12}, {U32Pp16}, and {U4Pp4-x} ions.

  6. Biosynthesis and possible functions of inositol pyrophosphates in plants

    PubMed Central

    Williams, Sarah P.; Gillaspy, Glenda E.; Perera, Imara Y.

    2015-01-01

    Inositol phosphates (InsPs) are intricately tied to lipid signaling, as at least one portion of the inositol phosphate signaling pool is derived from hydrolysis of the lipid precursor, phosphatidyl inositol (4,5) bisphosphate. The focus of this review is on the inositol pyrophosphates, which are a novel group of InsP signaling molecules containing diphosphate or triphosphate chains (i.e., PPx) attached to the inositol ring. These PPx-InsPs are emerging as critical players in the integration of cellular metabolism and stress signaling in non-plant eukaryotes. Most eukaryotes synthesize the precursor molecule, myo-inositol (1,2,3,4,5,6)-hexakisphosphate (InsP6), which can serve as a signaling molecule or as storage compound of inositol, phosphorus, and minerals (referred to as phytic acid). Even though plants produce huge amounts of precursor InsP6 in seeds, almost no attention has been paid to whether PPx-InsPs exist in plants, and if so, what roles these molecules play. Recent work has delineated that Arabidopsis has two genes capable of PP-InsP5 synthesis, and PPx-InsPs have been detected across the plant kingdom. This review will detail the known roles of PPx-InsPs in yeast and animal systems, and provide a description of recent data on the synthesis and accumulation of these novel molecules in plants, and potential roles in signaling. PMID:25729385

  7. Differential diagnosis between secondary hyperparathyroidism and aluminum intoxication in uremic patients: Usefulness of /sup 99m/Tc-pyrophosphate bone scintigraphy

    SciTech Connect

    Kinnaert, P.; Van Hooff, I.; Schoutens, A.; Bergmann, P.; Fuss, M.; Dratwa, M.; Vienne, A.; Pasteels, J.L.; van Geertruyden, J.; Vanherweghem, J.L.

    1989-03-01

    Forty-one patients in chronic end-stage renal failure and 4 patients with a functioning kidney transplant presented with spontaneous hypercalcemia or intolerance to vitamin D3 sterols and/or oral calcium supplements. Bone iliac crest biopsy with aluminum staining and Tc-pyrophosphate bone scintigraphy with determination of Fogelman score were performed in all cases. Two patients had aluminum-induced osteomalacia (AL O). Thirty-eight biopsies showed renal osteodystrophy (secondary hyperparathyroidism or various combinations of osteitis fibrosa and osteomalacia): 19 with positive staining for aluminum (RO + AL) and 19 without aluminum deposits (RO). The series also comprised 2 cases of pure osteomalacia (OM), 2 cases of osteoporosis (OP), and 1 case of osteoporosis with aluminum accumulation (OP + AL). Mean Fogelman score in RO patients (9.1 +/- 0.3) was significantly higher than in all other categories (5.9 +/- 0.5 for RO + AL, and scores ranging from 0 to 8 in the last 7 patients, p less than 0.01). Patients with massive aluminum accumulation in bone (greater than 75% of the total trabecular surface) showed no or very low uptake of the isotope by the skeleton. Fogelman scores of 9 or higher were always associated with histological secondary hyperparathyroidism. /sup 99m/Tc-pyrophosphate bone scintigraphy is helpful to distinguish aluminum intoxication from secondary hyperparathyroidism in uremic patients.

  8. Nucleation and morphology of sodium metaborate dihydrate from NaOH solution

    NASA Astrophysics Data System (ADS)

    Qin, Shiyue; Zhang, Yifei; Zhang, Yi

    2016-01-01

    Szaibelyite ore is an important boron mineral used for producing boron compounds. Sodium metaborate dihydrate can be prepared through leaching of the szaibelyite ore in NaOH solution and the leaching liquor mainly consists of NaBO2 and NaOH. In this work, the induction time for sodium metaborate dihydrate crystallized in NaOH solution from 30 to 50 °C was systematically investigated. The primary nucleation and growth mechanism were determined on the basis of the induction time measurements. The crystals of various morphologies under different crystallization conditions were obtained: the rod-like crystals preferred to form at low temperature, while the plate-like crystals formed at high temperature; when the crystallization temperature was 30 °C, the flat rod-like crystals formed at low supersaturation, while the slim rod-like crystals formed at high supersaturation. Finally, the growth mechanism of the sodium metaborate dihydrate was identified with various models and the 2D nucleation-mediated model gave satisfactory fitting results.

  9. Functional Characterization of the Xanthophyllomyces dendrorhous Farnesyl Pyrophosphate Synthase and Geranylgeranyl Pyrophosphate Synthase Encoding Genes That Are Involved in the Synthesis of Isoprenoid Precursors

    PubMed Central

    Niklitschek, Mauricio; Sepúlveda, Dionisia; Rojas, María Cecilia; Baeza, Marcelo; Cifuentes, Víctor

    2014-01-01

    The yeast Xanthophyllomyces dendrorhous synthesizes the carotenoid astaxanthin, which has applications in biotechnology because of its antioxidant and pigmentation properties. However, wild-type strains produce too low amounts of carotenoids to be industrially competitive. Considering this background, it is indispensable to understand how the synthesis of astaxanthin is controlled and regulated in this yeast. In this work, the steps leading to the synthesis of the carotenoid precursor geranylgeranyl pyrophosphate (GGPP, C20) in X. dendrorhous from isopentenyl pyrophosphate (IPP, C5) and dimethylallyl pyrophosphate (DMAPP, C5) was characterized. Two prenyl transferase encoding genes, FPS and crtE, were expressed in E. coli. The enzymatic assays using recombinant E. coli protein extracts demonstrated that FPS and crtE encode a farnesyl pyrophosphate (FPP, C15) synthase and a GGPP-synthase, respectively. X. dendrorhous FPP-synthase produces geranyl pyrophosphate (GPP, C10) from IPP and DMAPP and FPP from IPP and GPP, while the X. dendrorhous GGPP-synthase utilizes only FPP and IPP as substrates to produce GGPP. Additionally, the FPS and crtE genes were over-expressed in X. dendrorhous, resulting in an increase of the total carotenoid production. Because the parental strain is diploid, the deletion of one of the alleles of these genes did not affect the total carotenoid production, but the composition was significantly altered. These results suggest that the over-expression of these genes might provoke a higher carbon flux towards carotenogenesis, most likely involving an earlier formation of a carotenogenic enzyme complex. Conversely, the lower carbon flux towards carotenogenesis in the deletion mutants might delay or lead to a partial formation of a carotenogenic enzyme complex, which could explain the accumulation of astaxanthin carotenoid precursors in these mutants. In conclusion, the FPS and the crtE genes represent good candidates to manipulate to favor

  10. Calcium oxalate monohydrate precipitation investigation by thermometric method

    NASA Astrophysics Data System (ADS)

    Söhnel, O.; Costa-Bauzá, A.; Velich, V.

    1993-01-01

    Calcium oxalate monohydrate (COM) precipitation from diluted solutions of 100 mol m -3 ionic strength at 25°C was studied by an isoperibolic reaction twin calorimeter. The molar reaction enthalpy was determined as - 17.5 kJ mol -1. Results achieved with a pure system were highly reproducible. Citrate, pyrophosphate and phytate retard COM precipitation that is manifested mainly by an induction period appearance and a decrease of the initial precipitation rate. Effect of the studied impurities on individual precipitation experiments carried out under identical conditions was to some extent "random", i.e. the reaction extent reached at arbitrary time considerably differed for individual experiments. Impurity effectiveness in retarding spontaneous precipitation increases in succession citrate < pyrophosphate < phytate.

  11. Farnesyl pyrophosphate synthase is the molecular target of nitrogen-containing bisphosphonates.

    PubMed

    van Beek, E; Pieterman, E; Cohen, L; Löwik, C; Papapoulos, S

    1999-10-14

    Bisphosphonates (Bps), inhibitors of osteoclastic bone resorption, are used in the treatment of skeletal disorders. Recent evidence indicated that farnesyl pyrophosphate (FPP) synthase and/or isopentenyl pyrophosphate (IPP) isomerase is the intracellular target(s) of bisphosphonate action. To examine which enzyme is specifically affected, we determined the effect of different Bps on incorporation of [(14)C]mevalonate (MVA), [(14)C]IPP, and [(14)C]dimethylallyl pyrophosphate (DMAPP) into polyisoprenyl pyrophosphates in a homogenate of bovine brain. HPLC analysis revealed that the three intermediates were incorporated into FPP and geranylgeranyl pyrophosphate (GGPP). In contrast to clodronate, the nitrogen-containing Bps (NBps), alendronate, risedronate, olpadronate, and ibandronate, completely blocked FPP and GGPP formation and induced in incubations with [(14)C]MVA a 3- to 5-fold increase in incorporation of label into IPP and/or DMAPP. Using a method that could distinguish DMAPP from IPP on basis of their difference in stability in acid, we found that none of the NBps affected the conversion of [(14)C]IPP into DMAPP, catalyzed by IPP isomerase, excluding this enzyme as target of NBp action. On the basis of these and our previous findings, we conclude that none of the enzymes up- or downstream of FPP synthase are affected by NBps, and FPP synthase is, therefore, the exclusive molecular target of NBp action.

  12. The biochemical properties of the mitochondrial thiamine pyrophosphate carrier from Drosophila melanogaster.

    PubMed

    Iacopetta, Domenico; Carrisi, Chiara; De Filippis, Giuseppina; Calcagnile, Valeria M; Cappello, Anna R; Chimento, Adele; Curcio, Rosita; Santoro, Antonella; Vozza, Angelo; Dolce, Vincenza; Palmieri, Ferdinando; Capobianco, Loredana

    2010-03-01

    The mitochondrial carriers are a family of transport proteins that shuttle metabolites, nucleotides and cofactors across the inner mitochondrial membrane. The genome of Drosophila melanogaster encodes at least 46 members of this family. Only five of these have been characterized, whereas the transport functions of the remainder cannot be assessed with certainty. In the present study, we report the functional identification of two D. melanogaster genes distantly related to the human and yeast thiamine pyrophosphate carrier (TPC) genes as well as the corresponding expression pattern throughout development. Furthermore, the functional characterization of the D. melanogaster mitochondrial thiamine pyrophosphate carrier protein (DmTpc1p) is described. DmTpc1p was over-expressed in bacteria, the purified protein was reconstituted into liposomes, and its transport properties and kinetic parameters were characterized. Reconstituted DmTpc1p transports thiamine pyrophosphate and, to a lesser extent, pyrophosphate, ADP, ATP and other nucleotides. The expression of DmTpc1p in Saccharomyces cerevisiaeTPC1 null mutant abolishes the growth defect on fermentable carbon sources. The main role of DmTpc1p is to import thiamine pyrophosphate into mitochondria by exchange with intramitochondrial ATP and/or ADP.

  13. Substrate binding mode and reaction mechanism of undecaprenyl pyrophosphate synthase deduced from crystallographic studies

    PubMed Central

    Chang, Sing-Yang; Ko, Tzu-Ping; Chen, Annie P.-C.; Wang, Andrew H.-J.; Liang, Po-Huang

    2004-01-01

    Undecaprenyl pyrophosphate synthase (UPPs) catalyzes eight consecutive condensation reactions of farnesyl pyrophosphate (FPP) with isopentenyl pyrophosphate (IPP) to form a 55-carbon long-chain product. We previously reported the crystal structure of the apo-enzyme from Escherichia coli and the structure of UPPs in complex with sulfate ions (resembling pyrophosphate of substrate), Mg2+, and two Triton molecules (product-like). In the present study, FPP substrate was soaked into the UPPs crystals, and the complex structure was solved. Based on the crystal structure, the pyrophosphate head group of FPP is bound to the backbone NHs of Gly29 and Arg30 as well as the side chains of Asn28, Arg30, and Arg39 through hydrogen bonds. His43 is close to the C2 carbon of FPP and may stabilize the farnesyl cation intermediate during catalysis. The hydrocarbon moiety of FPP is bound with hydrophobic amino acids including Leu85, Leu88, and Phe89, located on the α3 helix. The binding mode of FPP in cis-type UPPs is apparently different from that of trans-type and many other prenyltransferases which utilize Asprich motifs for substrate binding via Mg2+. The new structure provides a plausible mechanism for the catalysis of UPPs. PMID:15044730

  14. Sensitivity of technetium-99m-pyrophosphate scintigraphy in diagnosing cardiac amyloidosis

    SciTech Connect

    Falk, R.H.; Lee, V.W.; Rubinow, A.; Hood, W.B. Jr.; Cohen, A.S.

    1983-03-01

    To determine the value of technetium-99m-pyrophosphate myocardial scintigraphy in the diagnosis of amyloid heart disease this procedure was prospectively performed in 20 consecutive patients with biopsy-proven primary amyloidosis. Eleven patients had echocardiographic abnormalities compatible with amyloid cardiomyopathy, 9 of whom had congestive heart failure. Diffuse myocardial pyrophosphate uptake was of equal or greater intensity than that of the ribs in 9 of the 11 patients with echocardiograms suggestive of amyloidosis, but in only 2 of the 9 with normal echocardiograms, despite abnormal electrocardiograms (p less than 0.01). Increased wall thickness measured by M-mode echocardiography correlated with myocardial pyrophosphate uptake (r . 0.68, p less than 0.01). None of 10 control patients with nonamyloid, nonischemic heart disease had a strongly positive myocardial pyrophosphate uptake. Thus, myocardial technetium-99m-pyrophosphate scanning is a sensitive and specific test for the diagnosis of cardiac amyloidosis in patients with congestive heart failure of obscure origin. It does not appear to be of value for the early detection of cardiac involvement in patients with known primary amyloidosis without echocardiographic abnormalities.

  15. Calcium and phosphate release from resin-based materials containing different calcium orthophosphate nanoparticles.

    PubMed

    Rodrigues, Marcela C; Natale, Livia C; Arana-Chaves, Victor E; Braga, Roberto R

    2015-11-01

    The study compared ion release from resin-based materials containing calcium orthophosphates. Amorphous calcium phosphate (ACP), dicalcium phosphate anhydrous (DCPA), dicalcium phosphate dihydrate (DCPD), and tricalcium phosphate (β-TCP) nanoparticles were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), dynamic light scattering (DLS), and surface area (nitrogen adsorption isotherms, BET method). Nanoparticles were added to a dimethacrylate-based resin and materials were tested for degree of conversion (DC) and calcium/phosphate release up to 28 days under pH 5.5 and 7.0. Data were analyzed by ANOVA/Tukey test (alpha: 0.05).The crystallinity of DCPA, DCPD, and β-TCP were confirmed, as well as the ACP amorphous nature. DCPD and β-TCP presented larger agglomerates than DCPA and ACP. The surface area of ACP was 5-11 times higher than those of the other nanoparticles. Materials showed similar DC. The material containing ACP released significantly more ions than the others, which released similar amounts of calcium and, in most cases, phosphate. Ion release was not affected by pH. Calcium release decreased between 7 and 21 days, while phosphate levels remained constant after 14 days. In conclusion, ACP higher ion release can be ascribed to its high surface area. DCPA, DCPD, and β-TCP had similar performances as ion-releasing fillers. © 2015 Wiley Periodicals, Inc.

  16. Farnesyl pyrophosphate regulates adipocyte functions as an endogenous PPARγ agonist.

    PubMed

    Goto, Tsuyoshi; Nagai, Hiroyuki; Egawa, Kahori; Kim, Young-Il; Kato, Sota; Taimatsu, Aki; Sakamoto, Tomoya; Ebisu, Shogo; Hohsaka, Takahiro; Miyagawa, Hiroh; Murakami, Shigeru; Takahashi, Nobuyuki; Kawada, Teruo

    2011-08-15

    The cholesterol biosynthetic pathway produces not only sterols but also non-sterol mevalonate metabolites involved in isoprenoid synthesis. Mevalonate metabolites affect transcriptional and post-transcriptional events that in turn affect various biological processes including energy metabolism. In the present study, we examine whether mevalonate metabolites activate PPARγ (peroxisome-proliferator-activated receptor γ), a ligand-dependent transcription factor playing a central role in adipocyte differentiation. In the luciferase reporter assay using both GAL4 chimaera and full-length PPARγ systems, a mevalonate metabolite, FPP (farnesyl pyrophosphate), which is the precursor of almost all isoprenoids and is positioned at branch points leading to the synthesis of other longer-chain isoprenoids, activated PPARγ in a dose-dependent manner. FPP induced the in vitro binding of a co-activator, SRC-1 (steroid receptor co-activator-1), to GST (glutathione transferase)-PPARγ. Direct binding of FPP to PPARγ was also indicated by docking simulation studies. Moreover, the addition of FPP up-regulated the mRNA expression levels of PPARγ target genes during adipocyte differentiation induction. In the presence of lovastatin, an HMG-CoA (3-hydroxy-3-methylglutaryl-CoA) reductase inhibitor, both intracellular FPP levels and PPARγ-target gene expressions were decreased. In contrast, the increase in intracellular FPP level after the addition of zaragozic acid, a squalene synthase inhibitor, induced PPARγ-target gene expression. The addition of FPP and zaragozic acid promotes lipid accumulation during adipocyte differentiation. These findings indicated that FPP might function as an endogenous PPARγ agonist and regulate gene expression in adipocytes. © The Authors Journal compilation © 2011 Biochemical Society

  17. Farnesyl pyrophosphate regulates adipocyte functions as an endogenous PPARγ agonist

    PubMed Central

    Goto, Tsuyoshi; Nagai, Hiroyuki; Egawa, Kahori; Kim, Young-Il; Kato, Sota; Taimatsu, Aki; Sakamoto, Tomoya; Ebisu, Shogo; Hohsaka, Takahiro; Miyagawa, Hiroh; Murakami, Shigeru; Takahashi, Nobuyuki; Kawada, Teruo

    2011-01-01

    The cholesterol biosynthetic pathway produces not only sterols but also non-sterol mevalonate metabolites involved in isoprenoid synthesis. Mevalonate metabolites affect transcriptional and post-transcriptional events that in turn affect various biological processes including energy metabolism. In the present study, we examine whether mevalonate metabolites activate PPARγ (peroxisome-proliferator-activated receptor γ), a ligand-dependent transcription factor playing a central role in adipocyte differentiation. In the luciferase reporter assay using both GAL4 chimaera and full-length PPARγ systems, a mevalonate metabolite, FPP (farnesyl pyrophosphate), which is the precursor of almost all isoprenoids and is positioned at branch points leading to the synthesis of other longer-chain isoprenoids, activated PPARγ in a dose-dependent manner. FPP induced the in vitro binding of a co-activator, SRC-1 (steroid receptor co-activator-1), to GST (glutathione transferase)–PPARγ. Direct binding of FPP to PPARγ was also indicated by docking simulation studies. Moreover, the addition of FPP up-regulated the mRNA expression levels of PPARγ target genes during adipocyte differentiation induction. In the presence of lovastatin, an HMG-CoA (3-hydroxy-3-methylglutaryl-CoA) reductase inhibitor, both intracellular FPP levels and PPARγ-target gene expressions were decreased. In contrast, the increase in intracellular FPP level after the addition of zaragozic acid, a squalene synthase inhibitor, induced PPARγ-target gene expression. The addition of FPP and zaragozic acid promotes lipid accumulation during adipocyte differentiation. These findings indicated that FPP might function as an endogenous PPARγ agonist and regulate gene expression in adipocytes. PMID:21605082

  18. Computational Insights into Binding of Bisphosphates to Farnesyl Pyrophosphate Synthase

    PubMed Central

    Ohno, K; Mori, K; Orita, M; Takeuchi, M

    2011-01-01

    Bisphosphonates (BPs) are the most widely used and effective treatment for osteoporosis and Paget's disease. Non-nitrogen containing BPs (non-N-BPs), namely etidronate, clodronate, tiludronate, as well as nitrogen-containing BPs (N-BPs), namely pamidronate, alendronate, ibandronate, risedronate, zoledronate and minodronate have been launched on the market to date. N-BPs act by inhibiting the enzyme farnesyl pyrophosphate synthase (FPPS), and several crystal structures of complexes between FPPS and N-BPs have been revealed. Understanding the physical basis of the binding between protein and small molecules is an important goal in both medicinal chemistry and structural biology. In this review, we analyze in detail the energetic basis of molecular recognition between FPPS and N-BPs. First, we summarize the interactions between ligands and proteins observed in N-BPs-FPPS complexes in the Protein Data Bank (PDB). Second, we present an interaction energy analysis on the basis of full quantum mechanical calculation of FPPS and N-BP complexes using the fragment molecular orbital (FMO) method. The FMO result revealed that not only hydrogen bond and electrostatic interaction but also CH-O and π-π interaction with FPPS are important for N-BP’s potency. Third, we describe a binding site analysis of FPPS on the basis of the inhomogeneous solvation theory which, by clustering the results from an explicit solvent molecular dynamics simulation (MD), is capable of describing the entropic and enthalpic contributions to the free energies of individual hydration sites. Finally, we also discuss the structure-activity relationship (SAR) of the series of minodronate derivatives. PMID:21110804

  19. Pathway of thiamine pyrophosphate synthesis in Micrococcus denitrificans.

    PubMed Central

    Sanemori, H; Egi, Y; Kawasaki, T

    1976-01-01

    The pathway of thiamine pyrophosphate (TPP) biosynthesis, which is formed either from exogeneously added thiamine or from the pyrimidine and thiazole moieties of thiamine, in Micrococcus denitrificans was investigated. The following indirect evidence shows that thiamine pyrophosphokinase (EC 2.7.6.2) catalyzes the synthesis of TPP from thiamine: (i) [35S]thiamine incubated with cells of this microorganism was detected in the form of [35S]thiamine; (ii) thiamine gave a much faster rate of TPP synthesis than thiamine monophosphate (TMP) when determined with the extracts; and (iii) a partially purified preparation of the extracts can use thiamine, but not TMP, as the substrate. The activities of the four enzymes involved in TMP synthesis from pyrimidine and thiazole moieties of thiamine were detected in the extracts of M. denitrificans. The extracts contained a high activity of the phosphatase, probably specific for TMP. After M. denitrificans cells were grown on a minimal medium containing 3 mM adenosine, which causes derepression of de novo thiamine biosynthesis in Escherichia coli, the activities of the four enzymes involved with TMP synthesis, the TMP phosphatase, and the thiamine pyrophosphokinase were enhanced two- to threefold. These results indicate that TPP is synthesized directly from thiamine without forming TMP as an intermediate and that de novo synthesis of TPP from the pyrimidine and thiazole moieties involves the formation of TMP, followed by hydrolysis to thiamine, which is then converted to TPP directly. Thus, the pathway of TPP synthesis from TMP synthesized de novo in M. denitrificans is different from that found in E. coli, in which TMP synthesized de novo is converted directly to TPP without producing thiamine. PMID:181359

  20. Central Role of Pyrophosphate in Acellular Cementum Formation

    PubMed Central

    Foster, Brian L.; Nagatomo, Kanako J.; Nociti, Francisco H.; Fong, Hanson; Dunn, Daisy; Tran, Anne B.; Wang, Wei; Narisawa, Sonoko; Millán, Jose Luis; Somerman, Martha J.

    2012-01-01

    Background Inorganic pyrophosphate (PPi) is a physiologic inhibitor of hydroxyapatite mineral precipitation involved in regulating mineralized tissue development and pathologic calcification. Local levels of PPi are controlled by antagonistic functions of factors that decrease PPi and promote mineralization (tissue-nonspecific alkaline phosphatase, Alpl/TNAP), and those that increase local PPi and restrict mineralization (progressive ankylosis protein, ANK; ectonucleotide pyrophosphatase phosphodiesterase-1, NPP1). The cementum enveloping the tooth root is essential for tooth function by providing attachment to the surrounding bone via the nonmineralized periodontal ligament. At present, the developmental regulation of cementum remains poorly understood, hampering efforts for regeneration. To elucidate the role of PPi in cementum formation, we analyzed root development in knock-out (−/−) mice featuring PPi dysregulation. Results Excess PPi in the Alpl−/− mouse inhibited cementum formation, causing root detachment consistent with premature tooth loss in the human condition hypophosphatasia, though cementoblast phenotype was unperturbed. Deficient PPi in both Ank and Enpp1−/− mice significantly increased cementum apposition and overall thickness more than 12-fold vs. controls, while dentin and cellular cementum were unaltered. Though PPi regulators are widely expressed, cementoblasts selectively expressed greater ANK and NPP1 along the root surface, and dramatically increased ANK or NPP1 in models of reduced PPi output, in compensatory fashion. In vitro mechanistic studies confirmed that under low PPi mineralizing conditions, cementoblasts increased Ank (5-fold) and Enpp1 (20-fold), while increasing PPi inhibited mineralization and associated increases in Ank and Enpp1 mRNA. Conclusions Results from these studies demonstrate a novel developmental regulation of acellular cementum, wherein cementoblasts tune cementogenesis by modulating local levels of PPi

  1. Temperature-dependent formation of NaCl dihydrate in levitated NaCl and sea salt aerosol particles.

    PubMed

    Peckhaus, Andreas; Kiselev, Alexei; Wagner, Robert; Duft, Denis; Leisner, Thomas

    2016-12-28

    Recent laboratory studies indicate that the hydrated form of crystalline NaCl is potentially important for atmospheric processes involving depositional ice nucleation on NaCl dihydrate particles under cirrus cloud conditions. However, recent experimental studies reported a strong discrepancy between the temperature intervals where the efflorescence of NaCl dihydrate has been observed. Here we report the measurements of the volume specific nucleation rate of crystalline NaCl in the aqueous solution droplets of pure NaCl suspended in an electrodynamic balance at constant temperature and humidity in the range from 250 K to 241 K. Based on these measurements, we derive the interfacial energy of crystalline NaCl dihydrate in a supersaturated NaCl solution and determined its temperature dependence. Taking into account both temperature and concentration dependence of nucleation rate coefficients, we explain the difference in the observed fractions of NaCl dihydrate reported in the previous studies. Applying the heterogeneous classical nucleation theory model, we have been able to reproduce the 5 K shift of the NaCl dihydrate efflorescence curve observed for the sea salt aerosol particles, assuming the presence of super-micron solid inclusions (hypothetically gypsum or hemihydrate of CaSO4). These results support the notion that the phase transitions in microscopic droplets of supersaturated solution should be interpreted by accounting for the stochastic nature of homogeneous and heterogeneous nucleation and cannot be understood on the ground of bulk phase diagrams alone.

  2. Temperature-dependent formation of NaCl dihydrate in levitated NaCl and sea salt aerosol particles

    NASA Astrophysics Data System (ADS)

    Peckhaus, Andreas; Kiselev, Alexei; Wagner, Robert; Duft, Denis; Leisner, Thomas

    2016-12-01

    Recent laboratory studies indicate that the hydrated form of crystalline NaCl is potentially important for atmospheric processes involving depositional ice nucleation on NaCl dihydrate particles under cirrus cloud conditions. However, recent experimental studies reported a strong discrepancy between the temperature intervals where the efflorescence of NaCl dihydrate has been observed. Here we report the measurements of the volume specific nucleation rate of crystalline NaCl in the aqueous solution droplets of pure NaCl suspended in an electrodynamic balance at constant temperature and humidity in the range from 250 K to 241 K. Based on these measurements, we derive the interfacial energy of crystalline NaCl dihydrate in a supersaturated NaCl solution and determined its temperature dependence. Taking into account both temperature and concentration dependence of nucleation rate coefficients, we explain the difference in the observed fractions of NaCl dihydrate reported in the previous studies. Applying the heterogeneous classical nucleation theory model, we have been able to reproduce the 5 K shift of the NaCl dihydrate efflorescence curve observed for the sea salt aerosol particles, assuming the presence of super-micron solid inclusions (hypothetically gypsum or hemihydrate of CaSO4). These results support the notion that the phase transitions in microscopic droplets of supersaturated solution should be interpreted by accounting for the stochastic nature of homogeneous and heterogeneous nucleation and cannot be understood on the ground of bulk phase diagrams alone.

  3. Human farnesyl pyrophosphate synthase is allosterically inhibited by its own product

    PubMed Central

    Park, Jaeok; Zielinski, Michal; Magder, Alexandr; Tsantrizos, Youla S.; Berghuis, Albert M.

    2017-01-01

    Farnesyl pyrophosphate synthase (FPPS) is an enzyme of the mevalonate pathway and a well-established therapeutic target. Recent research has focused around a newly identified druggable pocket near the enzyme's active site. Pharmacological exploitation of this pocket is deemed promising; however, its natural biological function, if any, is yet unknown. Here we report that the product of FPPS, farnesyl pyrophosphate (FPP), can bind to this pocket and lock the enzyme in an inactive state. The Kd for this binding is 5–6 μM, within a catalytically relevant range. These results indicate that FPPS activity is sensitive to the product concentration. Kinetic analysis shows that the enzyme is inhibited through FPP accumulation. Having a specific physiological effector, FPPS is a bona fide allosteric enzyme. This allostery offers an exquisite mechanism for controlling prenyl pyrophosphate levels in vivo and thus contributes an additional layer of regulation to the mevalonate pathway. PMID:28098152

  4. The protective effect of thiamine pyrophosphate, but not thiamine, against cardiotoxicity induced with cisplatin in rats.

    PubMed

    Coskun, Resit; Turan, Mehmet Ibrahim; Turan, Isil Siltelioglu; Gulapoglu, Mine

    2014-07-01

    This study investigated the effect of thiamine pyrophosphate on oxidative damage associated with cardiotoxicity caused by cisplatin (CIS), an antineoplastic agent, in rats, and compared this with thiamine. Animals used in the study were divided into four groups of 6 rats each. These represented a control group receiving 5 mg/kg of CIS, study groups receiving 20 mg/kg of thiamine pyrophosphate plus 5 mg/kg of cisplatin (CTPG) or 20 mg/kg of thiamine plus 5 mg/kg of cisplatin and a healthy (H) group. All doses were administered intraperitoneally once a day for 14 days. Malondialdehyde, total glutathione and products of DNA injury results were similar in the CTPG and H groups (p > 0.05). Creatinine kinase, creatine kinase MB and troponin 1 levels were similar in the CTPG and H groups (p > 0.05). Thiamine pyrophosphate prevented CIS-associated oxidative stress and heart injury, whereas thiamine did not prevent these.

  5. Synthesis of high specific activity (1- sup 3 H) farnesyl pyrophosphate

    SciTech Connect

    Saljoughian, M.; Morimoto, H.; Williams, P.G.

    1991-08-01

    The synthesis of tritiated farnesyl pyrophosphate with high specific activity is reported. trans-trans Farnesol was oxidized to the corresponding aldehyde followed by reduction with lithium aluminium tritide (5%-{sup 3}H) to give trans-trans (1-{sup 3}H)farnesol. The specific radioactivity of the alcohol was determined from its triphenylsilane derivative, prepared under very mild conditions. The tritiated alcohol was phosphorylated by initial conversion to an allylic halide, and subsequent treatment of the halide with tris-tetra-n-butylammonium hydrogen pyrophosphate. The hydride procedure followed in this work has advantages over existing methods for the synthesis of tritiated farnesyl pyrophosphate, with the possibility of higher specific activity and a much higher yield obtained. 10 refs., 3 figs.

  6. Applying Molecular Dynamics Simulations to Identify Rarely Sampled Ligand-bound Conformational States of Undecaprenyl Pyrophosphate Synthase, an Antibacterial Target

    SciTech Connect

    Sinko, William; de Oliveira, César; Williams, Sarah; Van Wynsberghe, Adam; Durrant, Jacob D.; Cao, Rong; Oldfield, Eric; McCammon, J. Andrew

    2012-04-30

    Undecaprenyl pyrophosphate synthase is a cis-prenyltransferase enzyme, which is required for cell wall biosynthesis in bacteria. Undecaprenyl pyrophosphate synthase is an attractive target for antimicrobial therapy. We performed long molecular dynamics simulations and docking studies on undecaprenyl pyrophosphate synthase to investigate its dynamic behavior and the influence of protein flexibility on the design of undecaprenyl pyrophosphate synthase inhibitors. We also describe the first X-ray crystallographic structure of Escherichia coli apo-undecaprenyl pyrophosphate synthase. The molecular dynamics simulations indicate that undecaprenyl pyrophosphate synthase is a highly flexible protein, with mobile binding pockets in the active site. By carrying out docking studies with experimentally validated undecaprenyl pyrophosphate synthase inhibitors using high- and low-populated conformational states extracted from the molecular dynamics simulations, we show that structurally dissimilar compounds can bind preferentially to different and rarely sampled conformational states. By performing structural analyses on the newly obtained apo-undecaprenyl pyrophosphate synthase and other crystal structures previously published, we show that the changes observed during the molecular dynamics simulation are very similar to those seen in the crystal structures obtained in the presence or absence of ligands. We believe that this is the first time that a rare 'expanded pocket' state, key to drug design and verified by crystallography, has been extracted from a molecular dynamics simulation.

  7. Calcium Test

    MedlinePlus

    ... Hyperthyroidism Sarcoidosis Tuberculosis Prolonged immobilization Excess vitamin D intake Thiazide diuretics Kidney transplant HIV/AIDS Low total calcium (hypocalcemia) The most common cause of low total calcium is: Low blood protein levels, especially a low level of albumin , which ...

  8. Isopentenyl Pyrophosphate cis-1,4-Polyisoprenyl Transferase from Guayule (Parthenium argentatum Gray) 1

    PubMed Central

    Madhavan, S.; Benedict, Chauncey R.

    1984-01-01

    Electron micrographs of the mesophyll cells of guayule Parthenium argentatum Gray leaves show deposits of cis-polyisoprene (rubber) in the cytoplasm in the vicinity of mitochondria and chloroplasts and demonstrate that the rubber-synthesizing enzymes are present in guayule leaves. The terminal step in the synthesis of cis-polyisoprene from isopentenyl pyrophosphate (IPP) catalyzed by isopentenyl pyrophosphate cis-1,4-polyisoprenyl transferase has been demonstrated in crude leaf extracts by the enzymic incorporation of [14C]isopentenyl pyrophosphate into the polymer and the recovery of [14C]levulinic acid following ozonolysis. The rubber transferase activity in the crude extracts of guayule leaves was 5.8 nanomoles isopentenyl pyrophosphate incorporated per milligram protein per hour. This is the first description of the rubber transferase from a nonlaticiferous plant. The specific activity (in units of nanomoles IPP converted per milligram protein per hour) of the partially purified enzyme following chromatography on diethylaminoethyl-cellulose columns was 41.7 units and contained 0.29 units of IPP isomerase activity and 0.08 units of farnesyl pyrophosphate synthetase activity. The rubber transferase requires reduced glutathione and Mg2+ for maximal activity. There was no incorporation of IPP into cis-1,4-polyisoprene in the absence of rubber particles as primer, and Langmuir isotherm plots showed that the specific activity of the enzyme was proportional to the concentration of the enzyme on the surface of the rubber particles. For a given rubber particle distribution, enzyme activity was proportional to time, IPP concentration, and rubber concentration. The addition of 0.4 millimolar dimethylallyl pyrophosphate to the rubber transferase reaction resulted in a 2-fold increase in the incorporation of IPP into rubber. A comparison was made of the relative activities of rubber transferase in different species of Parthenium, Ficus, and Euphorbia. Images Fig. 2 Fig. 3

  9. Pharmacokinetics of Ferric Pyrophosphate Citrate, a Novel Iron Salt, Administered Intravenously to Healthy Volunteers.

    PubMed

    Pratt, Raymond D; Swinkels, Dorine W; Ikizler, T Alp; Gupta, Ajay

    2017-03-01

    Ferric pyrophosphate citrate (Triferic) is a water-soluble iron salt that is administered via dialysate to maintain iron balance and hemoglobin in hemodialysis patients. This double-blind, randomized, placebo-controlled, single-, ascending-dose study was conducted to evaluate the pharmacokinetics and safety of intravenous ferric pyrophosphate citrate in 48 healthy iron-replete subjects (drug, n = 36; placebo, n = 12). Single doses of 2.5, 5.0, 7.5, or 10 mg of ferric pyrophosphate citrate or placebo were administered over 4 hours, and single doses of 15 or 20 mg of ferric pyrophosphate citrate or placebo were administered over 12 hours via intravenous infusion. Serum total iron (sFetot ), transferrin-bound iron (TBI), hepcidin-25, and biomarkers of oxidative stress and inflammation were determined using validated assays. Marked diurnal variation in sFetot was observed in placebo-treated subjects. Concentrations of sFetot and TBI increased rapidly after drug administration, with maximum serum concentrations (Cmax ) reached at the end of infusion. Increases in baseline-corrected Cmax and area under the concentration-time curve from 0 to the time of the last quantifiable concentration (AUC0-t ) were dose proportional up to 100% transferrin saturation. Iron was rapidly cleared (apparent terminal phase half-life 1.2-2 hours). No significant changes from baseline in serum hepcidin-25 concentration were observed at end of infusion for any dose. Biomarkers of oxidative stress and inflammation were unaffected. Intravenous doses of ferric pyrophosphate citrate were well tolerated. These results demonstrate that intravenous ferric pyrophosphate citrate is rapidly bound to transferrin and cleared from the circulation without increasing serum hepcidin levels or biomarkers of oxidative stress or inflammation. © 2016 The Authors. The Journal of Clinical Pharmacology Published by Wiley Periodicals, Inc. on behalf of American College of Clinical Pharmacology.

  10. Isopentenyl Pyrophosphate cis-1,4-Polyisoprenyl Transferase from Guayule (Parthenium argentatum Gray).

    PubMed

    Madhavan, S; Benedict, C R

    1984-08-01

    Electron micrographs of the mesophyll cells of guayule Parthenium argentatum Gray leaves show deposits of cis-polyisoprene (rubber) in the cytoplasm in the vicinity of mitochondria and chloroplasts and demonstrate that the rubber-synthesizing enzymes are present in guayule leaves. The terminal step in the synthesis of cis-polyisoprene from isopentenyl pyrophosphate (IPP) catalyzed by isopentenyl pyrophosphate cis-1,4-polyisoprenyl transferase has been demonstrated in crude leaf extracts by the enzymic incorporation of [(14)C]isopentenyl pyrophosphate into the polymer and the recovery of [(14)C]levulinic acid following ozonolysis. The rubber transferase activity in the crude extracts of guayule leaves was 5.8 nanomoles isopentenyl pyrophosphate incorporated per milligram protein per hour. This is the first description of the rubber transferase from a nonlaticiferous plant.The specific activity (in units of nanomoles IPP converted per milligram protein per hour) of the partially purified enzyme following chromatography on diethylaminoethyl-cellulose columns was 41.7 units and contained 0.29 units of IPP isomerase activity and 0.08 units of farnesyl pyrophosphate synthetase activity. The rubber transferase requires reduced glutathione and Mg(2+) for maximal activity. There was no incorporation of IPP into cis-1,4-polyisoprene in the absence of rubber particles as primer, and Langmuir isotherm plots showed that the specific activity of the enzyme was proportional to the concentration of the enzyme on the surface of the rubber particles. For a given rubber particle distribution, enzyme activity was proportional to time, IPP concentration, and rubber concentration. The addition of 0.4 millimolar dimethylallyl pyrophosphate to the rubber transferase reaction resulted in a 2-fold increase in the incorporation of IPP into rubber. A comparison was made of the relative activities of rubber transferase in different species of Parthenium, Ficus, and Euphorbia.

  11. Pharmacokinetics of Ferric Pyrophosphate Citrate, a Novel Iron Salt, Administered Intravenously to Healthy Volunteers

    PubMed Central

    Swinkels, Dorine W.; Ikizler, T. Alp; Gupta, Ajay

    2016-01-01

    Abstract Ferric pyrophosphate citrate (Triferic) is a water‐soluble iron salt that is administered via dialysate to maintain iron balance and hemoglobin in hemodialysis patients. This double‐blind, randomized, placebo‐controlled, single‐, ascending‐dose study was conducted to evaluate the pharmacokinetics and safety of intravenous ferric pyrophosphate citrate in 48 healthy iron‐replete subjects (drug, n = 36; placebo, n = 12). Single doses of 2.5, 5.0, 7.5, or 10 mg of ferric pyrophosphate citrate or placebo were administered over 4 hours, and single doses of 15 or 20 mg of ferric pyrophosphate citrate or placebo were administered over 12 hours via intravenous infusion. Serum total iron (sFetot), transferrin‐bound iron (TBI), hepcidin‐25, and biomarkers of oxidative stress and inflammation were determined using validated assays. Marked diurnal variation in sFetot was observed in placebo‐treated subjects. Concentrations of sFetot and TBI increased rapidly after drug administration, with maximum serum concentrations (Cmax) reached at the end of infusion. Increases in baseline‐corrected Cmax and area under the concentration‐time curve from 0 to the time of the last quantifiable concentration (AUC0‐t) were dose proportional up to 100% transferrin saturation. Iron was rapidly cleared (apparent terminal phase half‐life 1.2‐2 hours). No significant changes from baseline in serum hepcidin‐25 concentration were observed at end of infusion for any dose. Biomarkers of oxidative stress and inflammation were unaffected. Intravenous doses of ferric pyrophosphate citrate were well tolerated. These results demonstrate that intravenous ferric pyrophosphate citrate is rapidly bound to transferrin and cleared from the circulation without increasing serum hepcidin levels or biomarkers of oxidative stress or inflammation. PMID:27557937

  12. Crystallization of calcium phosphate in polyacrylamide hydrogels containing phosphate ions

    NASA Astrophysics Data System (ADS)

    Yokoi, Taishi; Kawashita, Masakazu; Kikuta, Koichi; Ohtsuki, Chikara

    2010-08-01

    Calcium phosphate crystals were formed in polyacrylamide (PAAm) hydrogels containing phosphate ions by diffusion of calcium ions from calcium nitrate (Ca(NO 3) 2) solutions covering the gels. Changes in crystalline phases and crystal morphology of calcium phosphate, and in ion concentrations of the Ca(NO 3) 2 solutions were investigated as a function of reaction time. Single or two coexisting crystalline phases of calcium phosphate, hydroxyapatite (HAp), HAp/dicalcium phosphate dihydrate (DCPD) or octacalcium phosphate (OCP)/DCPD were formed in the gels. HAp crystals are formed near the surface of the gels. The dense HAp layer and HAp/DCPD layer prevented diffusion of calcium ions from the Ca(NO 3) 2 solution, thus formation of calcium phosphate in the gel phase was inhibited. Formation of DCPD was observed to follow the formation of OCP or HAp. The size of the OCP crystals gradually increased with reaction time, while changes in size of HAp crystals were not observed. The reaction time required for DCPD formation depended on the degree of supersaturation with respect to DCPD in the systems. DCPD formed within 1 day under high supersaturation conditions, whereas it formed at 10 days in low supersaturation conditions.

  13. Thallium-201 and technetium-99m-pyrophosphate: myocardial imaging in the coronary care unit

    SciTech Connect

    Wackers, F.J.T.

    1980-01-01

    The editor and contributors have developed a textbook that explains the use of thallium 201 and technetium-99m pyrophosphate for the detection of acute myocardial infarct. Their integrated approach emphasizes the complementary nature of both imaging modalities. One aim of this book is to discuss the technical characteristics of the two myocardial imaging methods. The text has fourteen chapters. Instrumentation, mechanisms of accumulation of thallium 201 and technetium-99m pyrophosphate, acute myocardial infarct, unstable angina pectoris, atypical chest pain, and the interrelationship of the techniques are among the topics discussed. (JMT)

  14. New synthesis routes for indium-doped tin pyrophosphate proton conductors

    SciTech Connect

    Garzon, Fernando H; Mukundan, Rangachary; Brosha, Eric L; Einsla, Melinda L

    2008-01-01

    We have synthesized various indium-tin pyrophosphates using the traditional high-temperature method described in the literature as well as a novel solution-precipitation process developed in our laboratory. An ammonium-based precursor was used in an attempt to achieve a lower crystallization temperature. The use of this precursor in the solution-precipitation of indium-tin pyrophosphate gels resulted in precursors that could be crystallized at less than 600 {sup o}C. Inelastic neutron scattering studies indicate the presence of phosphate rich grain boundary phases that may be responsible for the high conductivity of the materials.

  15. Technetium-99m pyrophosphate imaging in acute renal failure associated with nontraumatic rhabdomyolysis

    SciTech Connect

    Patel, R.; Mishkin, F.S.

    1986-10-01

    Technetium-99m pyrophosphate (Tc-PYP) imaging was performed in five patients with acute renal failure associated with nontraumatic rhabdomyolysis. Four patients had phencyclidine intoxication and one had viral pneumonia. During the acute phase, marked uptake of pyrophosphate was seen in all patients in several muscle groups, but always in the thigh adductors. The results show that phencyclidine intoxication can result in diffuse muscle uptake of Tc-PYP without overt evidence of muscle injury. Tc-PYP imaging may provide a clue to the cause of acute renal failure in patients with suspected rhabdomyolysis in whom elevations of serum creatine phosphokinase concentrations are equivocal.

  16. Biosynthesis of monoterpenes. Enantioselectivity in the enzymatic cyclization of (+)- and (-)-linalyl pyrophosphate to (+)- and (-)-pinene and (+)- and (-)-camphene

    SciTech Connect

    Croteau, R.; Satterwhite, D.M.; Cane, D.E.; Chang, C.C.

    1988-07-25

    Cyclase I from Salvia officinalis leaf catalyzes the conversion of geranyl pyrophosphate to the stereo-chemically related bicyclic monoterpenes (+)-alpha-pinene and (+)-camphene and to lesser quantities of monocyclic and acyclic olefins, whereas cyclase II from this plant tissue converts the same acyclic precursor to (-)-alpha-pinene, (-)-beta-pinene and (-)-camphene as well as to lesser amounts of monocyclics and acyclics. These antipodal cyclizations are considered to proceed by the initial isomerization of the substrate to the respective bound tertiary allylic intermediates (-)-(3R)- and (+)-(3S)-linalyl pyrophosphate. ((3R)-8,9-14C,(3RS)-1E-3H)Linalyl pyrophosphate (3H:14C = 5.14) was tested as a substrate with both cyclases to determine the configuration of the cyclizing intermediate. This substrate with cyclase I yielded alpha-pinene and camphene with 3H:14C ratios of 3.1 and 4.2, respectively, indicating preferential, but not exclusive, utilization of the (3R)-enantiomer. With cyclase II, the doubly labeled substrate gave bicyclic olefins with 3H:14C ratios of from 13 to 20, indicating preferential, but not exclusive, utilization of the (3S)-enantiomer in this case. (3R)- and (3S)-(1Z-3H)linalyl pyrophosphate were separately compared to the achiral precursors (1-3H)geranyl pyrophosphate and (1-3H)neryl pyrophosphate (cis-isomer) as substrates for the cyclizations. With cyclase I, geranyl, neryl, and (3R)-linalyl pyrophosphate gave rise exclusively to (+)-alpha-pinene and (+)-camphene, whereas (3S)-linayl pyrophosphate produced, at relatively low rates, the (-)-isomers. With cyclase II, geranyl, neryl, and (3S)-linalyl pyrophosphate yielded exclusively the (-)-isomer series, whereas (3R)-linalyl pyrophosphate afforded the (+)-isomers at low rates.

  17. Hydrogen bonding. Part 26. Thermodynamics of dissociation and infrared spectracrystal structure correlations for betaine monohydrate and trimethylamine oxide dihydrate

    NASA Astrophysics Data System (ADS)

    Toccalino, Patricia L.; Harmon, Kenneth M.; Harmon, Jennifer

    1988-10-01

    Thermodynamic parameters for the dissociation of betaine monohydrate and trimethylamine oxide dihydrate have been determined by equilibrium vapor pressure measurements. Betaine monohydrate appears in two slightly different crystalline forms, one obtained by crystallization from water and the other by addition of water vapor to solid anhydrous betaine. Hydrogen bond energies in these hydrates are at least 8-9 kcal mol -1 per OH⋯O bond. Hydrogen bond energies in trimethylamine oxide dihydrate average at least 14 kcal mol -1 per OH⋯O bond; however, as there are two distinct types of hydrogen bonds in this hydrate, some bonds are stronger and some weaker than 14 kcal mol -1. These studies show conclusively that trimethylamine oxide monohydrate does not exist. The infrared spectrum of trimethylamine oxide dihydrate is correlated with the crystal structure.

  18. Postcountershock myocardial damage after pretreatment with adrenergic and calcium channel antagonists in halothane-anesthetized dogs

    SciTech Connect

    Gaba, D.M.; Metz, S.; Maze, M.

    1985-05-01

    Transthoracic electric countershock can cause necrotic myocardial lesions in humans as well as experimental animals. The authors investigated the effect on postcountershock myocardial damage of pretreatment with prazosin, an alpha-1 antagonist; L-metoprolol, a beta-1 antagonist, and verapamil, a calcium channel-blocking agent. Twenty dogs were anesthetized with halothane and given two transthoracic countershocks of 295 delivered joules each after drug or vehicle treatment. Myocardial injury was quantitated 24 h following countershock by measuring the uptake of technetium-99m pyrophosphate in the myocardium. Elevated technetium-99m pyrophosphate uptake occurred in visible lesions in most dogs regardless of drug treatment. For each of four parameters of myocardial damage there was no statistically significant difference between control animals and those treated with prazosin, metoprolol, or verapamil. These data suggest that adrenergic or calcium channel-mediated mechanisms are not involved in the pathogenesis of postcountershock myocardial damage.

  19. Zinc(II) and cadmium(II) metal complexes of thiamine pyrophosphate and 2-(alpha-hydroxyethyl)thiamine pyrophosphate: models for activation of pyruvate decarboxylase.

    PubMed

    Malandrinos, G; Louloudi, M; Koukkou, A I; Sovago, I; Drainas, C; Hadjiliadis, N

    2000-04-01

    Metal complexes of thiamine pyrophosphate (TPP) of the general formula [M2(TPPH)2Cl2]x4H2O (M = Zn2+, Cd2+) were isolated from methanolic solutions and characterized by elemental analysis, FT-IR, and multinuclear NMR spectroscopies. The data provide evidence for the bonding of the metals to the N(1') atom of the pyrimidine ring and to the pyrophosphate group. The stability constant measurements of TPP and 2-(alpha-hydroxyethyl)thiamine pyrophosphate (HETPP) metal complexes in aqueous solution imply the formation of dimeric complex species similar to the isolated solid products. They indicate also that HETPP forms more stable metal complexes than does TPP. To evaluate the coenzyme action of TPP and HETPP metal complexes, enzymic studies have been done using pyruvate decarboxylase apoenzyme. TPP metal complexes do not bind to the apoenzyme, unlike the Zn(II)-HETPP complex which can act as coenzyme. Considering these results, possible functional implications for thiamine involvement in catalysis are discussed.

  20. Acute and 3-month effects of calcium carbonate on the calcification propensity of serum and regulators of vascular calcification: secondary analysis of a randomized controlled trial.

    PubMed

    Bristow, S M; Gamble, G D; Pasch, A; O'Neill, W C; Stewart, A; Horne, A M; Reid, I R

    2016-03-01

    Calcium supplements have been associated with increased cardiovascular risk, but the mechanism is unknown. We investigated the effects of calcium supplements on the propensity of serum to calcify, based on the transition time of primary to secondary calciprotein particles (T50). Changes in serum calcium were related to changes in T50. Calcium supplements have been associated with increased cardiovascular risk; however, it is unknown whether this is related to an increase in vascular calcification. We investigated the acute and 3-month effects of calcium supplements on the propensity of serum to calcify, based on the transition time of primary to secondary calciprotein particles (T50), and on three possible regulators of calcification: fetuin-A, pyrophosphate and fibroblast growth factor-23 (FGF23). We randomized 41 postmenopausal women to 1 g/day of calcium as carbonate, or to a placebo containing no calcium. Measurements were performed at baseline and then 4 and 8 h after their first dose, and after 3 months of supplementation. Fetuin-A, pyrophosphate and FGF23 were measured in the first 10 participants allocated to calcium carbonate and placebo who completed the study. T50 declined in both groups, the changes tending to be greater in the calcium group. Pyrophosphate declined from baseline in the placebo group at 4 h and was different from the calcium group at this time point (p = 0.04). There were no other significant between-groups differences. The changes in serum total calcium from baseline were significantly related to changes in T50 at 4 h (r = -0.32, p = 0.05) and 8 h (r = -0.39, p = 0.01), to fetuin-A at 3 months (r = 0.57, p = 0.01) and to pyrophosphate at 4 h (r = 0.61, p = 0.02). These correlative findings suggest that serum calcium concentrations modulate the propensity of serum to calcify (T50), and possibly produce counter-regulatory changes in pyrophosphate and fetuin-A. This provides a possible mechanism by which

  1. Total lattice potential energy of sodium bromide dihydrate NaBr · 2H 2O

    NASA Astrophysics Data System (ADS)

    Herzig, P.; Jenkins, H. D. B.; Pritchett, M. S. F.

    1984-08-01

    In addition to presenting comparative calculations by two approaches for the total lattice potential energy of sodium bromide dihydrate, NaBr · 2H 2O, found to take the value 803.9 kJ mol -1, we investigate the influence of the size and nature of the basis set used to generate multipole moments in a Hartee-Fock calculation which are in turn used to calculate the Madelung constant. The requirement is one of critical size of the basis set and once this is reached the electrostatic energy will be reliable.

  2. Pseudopolymorphism in hydroxybenzophenones: the dihydrate of 2,2',4,4'-tetrahydroxybenzophenone.

    PubMed

    Landre, I M R; Martins, F T; Ellena, J A; Dos Santos, M H; Doriguetto, A C

    2012-04-01

    A dihydrate pseudopolymorph of bis(2,4-dihydroxyphenyl)methanone, C(13)H(10)O(5)·2H(2)O, (I), was obtained during polymorphism screening of hydroxybenzophenone derivatives. This structure, in which the molecule sits on a twofold axis, was compared with the known anhydrous form of (I) [Schlemper (1982). Acta Cryst. B38, 554-559]. The role of water in the crystal assembly was established on the basis of the known monohydrate pseudopolymorph of 3,4-dihydroxybenzophenone [Landre, Souza, Corrêa, Martins & Doriguetto (2010). Acta Cryst. C66, o463-o465].

  3. Thermal and dielectric properties of gel-grown cobalt malonate dihydrate single crystals

    NASA Astrophysics Data System (ADS)

    Mathew, Varghese; Xavier, Lizymol; Mahadevan, C. K.; Abraham, K. E.

    2011-03-01

    Single crystals of cobalt malonate dihydrate were grown in a silica gel medium by the single diffusion method. The thermal behavior of the crystals was investigated by thermogravimetric and differential thermal analyses. The dielectric constant, dielectric loss and ac conductivity of the crystals were estimated as a function of temperature in the range 40-140 °C for four different frequencies. The results indicate that the grown crystals are thermally stable up to about 150 °C and exhibit a phase transition at 130 °C.

  4. Nitric acid dihydrate at ambient and high pressure: An experimental and computational study

    SciTech Connect

    Walker, Martin; Pulham, Colin R.; Morrison, Carole A.; Allan, David R.; Marshall, William G.

    2006-06-01

    The high pressure structural behavior of nitric acid dihydrate ([H{sub 3}O]{sup +}{center_dot}[NO{sub 3}]{sup -}{center_dot}H{sub 2}O) has been investigated up to 3.8 GPa using single crystal x-ray diffraction and neutron powder diffraction techniques. A new structural phase has been identified above 1.33 GPa and this has been further studied by ab initio quantum mechanical calculations. These have guided the refinement by neutron powder diffraction.

  5. Dosimetric and kinetic investigations of γ-irradiated sodium tartrate dihydrate.

    PubMed

    Tuner, H; Kayikçi, M A

    2012-03-01

    Effects of gamma radiation on solid sodium tartrate dihydrate (NaTA) were studied using electron spin resonance (ESR) spectroscopy. One main singlet located at g = 2.0034 and many weak lines located at low and high magnetic field sides were found in the irradiated samples. Dosimetric and kinetic features of the radical species responsible for the experimental ESR spectra were explored through the variations in the signal intensities with respect to applied microwave power, temperature and storage time. Activation energies of the involved radical species were also determined using data derived from annealing studies.

  6. Fabrication of Poly-l-lactic Acid/Dicalcium Phosphate Dihydrate Composite Scaffolds with High Mechanical Strength—Implications for Bone Tissue Engineering

    PubMed Central

    Tanataweethum, Nida; Liu, Wai Ching; Scott Goebel, W.; Li, Ding; Chu, Tien Min

    2015-01-01

    Scaffolds were fabricated from poly-l-lactic acid (PLLA)/dicalcium phosphate dihydrate (DCPD) composite by indirect casting. Sodium citrate and PLLA were used to improve the mechanical properties of the DCPD scaffolds. The resulting PLLA/DCPD composite scaffold had increased diametral tensile strength and fracture energy when compared to DCPD only scaffolds (1.05 vs. 2.70 MPa and 2.53 vs. 12.67 N-mm, respectively). Sodium citrate alone accelerated the degradation rate by 1.5 times independent of PLLA. Cytocompatibility of all samples were evaluated using proliferation and differentiation parameters of dog-bone marrow stromal cells (dog-BMSCs). The results showed that viable dog-BMSCs attached well on both DCPD and PLLA/DCPD composite surfaces. In both DCPD and PLLA/DCPD conditioned medium, dog-BMSCs proliferated well and expressed alkaline phosphatase (ALP) activity indicating cell differentiation. These findings indicate that incorporating both sodium citrate and PLLA could effectively improve mechanical strength and biocompatibility without increasing the degradation time of calcium phosphate cement scaffolds for bone tissue engineering purposes. PMID:26556380

  7. Fabrication of Poly-l-lactic Acid/Dicalcium Phosphate Dihydrate Composite Scaffolds with High Mechanical Strength-Implications for Bone Tissue Engineering.

    PubMed

    Tanataweethum, Nida; Liu, Wai Ching; Goebel, W Scott; Li, Ding; Chu, Tien Min

    2015-11-04

    Scaffolds were fabricated from poly-l-lactic acid (PLLA)/dicalcium phosphate dihydrate (DCPD) composite by indirect casting. Sodium citrate and PLLA were used to improve the mechanical properties of the DCPD scaffolds. The resulting PLLA/DCPD composite scaffold had increased diametral tensile strength and fracture energy when compared to DCPD only scaffolds (1.05 vs. 2.70 MPa and 2.53 vs. 12.67 N-mm, respectively). Sodium citrate alone accelerated the degradation rate by 1.5 times independent of PLLA. Cytocompatibility of all samples were evaluated using proliferation and differentiation parameters of dog-bone marrow stromal cells (dog-BMSCs). The results showed that viable dog-BMSCs attached well on both DCPD and PLLA/DCPD composite surfaces. In both DCPD and PLLA/DCPD conditioned medium, dog-BMSCs proliferated well and expressed alkaline phosphatase (ALP) activity indicating cell differentiation. These findings indicate that incorporating both sodium citrate and PLLA could effectively improve mechanical strength and biocompatibility without increasing the degradation time of calcium phosphate cement scaffolds for bone tissue engineering purposes.

  8. ESR spectra of VO2+ ions adsorbed on calcium phosphates.

    PubMed

    Oniki, T; Doi, Y

    1983-07-01

    The ESR spectra of oxovanadium(IV) ions, (VO2+), adsorbed on hydroxyapatite(OHAp), fluorhydroxyapatite(FHAp), Mg-containing tricalcium phosphate(Mg-TCP), .octacalcium phosphate (OCP), dicalcium phosphate dihydrate (DCPD), and amorphous calcium phosphate(ACP) were measured at room temperature. The ESR parameters of VO2+ adsorbed on these compounds were slightly different from one another and accordingly, the ESR technique by use of VO2+ was useful for an analysis of the calcium phosphates precipitated from supersaturated solutions. The ESR parameters of VO2+ adsorbed on ACP and Mg-TCP were found to be very similar to each other, suggesting that ACP and TCP resemble each other in the structure of their crystal surfaces.

  9. Unexpectedly facile synthesis of symmetrical P1,P2-dinucleoside-5'pyrophosphates

    NASA Technical Reports Server (NTRS)

    Kanavarioti, Anastassia; Lu, Jonathan; Rosenbach, Morgan T.; Hurley, T. B.

    1991-01-01

    Symmetrical dinucleoside 5'-pyrophosphates have been synthesized from the corresponding nucleoside 5'-phosphate free acid in high yield. The one-pot procedure is carried out in DMF or DMSO using triphenylphosphine and 2,2'-dipyridyldisulfide as the coupling agents, and 1-methylimidazole as the catalyst.

  10. Modeling studies with Helicobacter pylori octaprenyl pyrophosphate synthase reveal the enzymatic mechanism of trans-prenyltransferases.

    PubMed

    Zhang, Jinyong; Zhang, Xiaoli; Zhang, Rui; Wu, Chao; Guo, Ying; Mao, Xuhu; Guo, Gang; Zhang, Ying; Wang, Da-Cheng; Li, Defeng; Zou, Quanming

    2012-12-01

    Octaprenyl pyrophosphate synthase (OPPs), an enzyme belonging to the trans-prenyltransferases family, is involved in the synthesis of C40 octaprenyl pyrophosphate (OPP) by reacting farnesyl pyrophosphate (FPP) with five isopentenyl pyrophosphates (IPP). It has been reported that OPPs is essential for bacteria's normal growth and is a potential target for novel antibacterial drug design. Here we report the crystal structure of OPPs from Helicobacter pylori, determined by MAD method at 2.8 Å resolution and refined to 2.0 Å resolution. The substrate IPP was docked into HpOPPs structure and residues involved in IPP recognition were identified. The other substrate FPP, the intermediate GGPP and a nitrogen-containing bisphosphonate drug were also modeled into the structure. The resulting model shed some lights on the enzymatic mechanism, including (1) residues Arg87, Lys36 and Arg39 are essential for IPP binding; (2) residues Lys162, Lys224 and Gln197 are involved in FPP binding; (3) the second DDXXD motif may involve in FPP binding by Mg(2+) mediated interactions; (4) Leu127 is probably involved in product chain length determination in HpOPPs and (5) the intermediate products such as GGPP need a rearrange to occupy the binding site of FPP and then IPP is reloaded. Our results also indicate that the nitrogen-containing bisphosphonate drugs are potential inhibitors of FPPs and other trans-prenyltransferases aiming at blocking the binding of FPP. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Unexpectedly facile synthesis of symmetrical P1,P2-dinucleoside-5'pyrophosphates

    NASA Technical Reports Server (NTRS)

    Kanavarioti, Anastassia; Lu, Jonathan; Rosenbach, Morgan T.; Hurley, T. B.

    1991-01-01

    Symmetrical dinucleoside 5'-pyrophosphates have been synthesized from the corresponding nucleoside 5'-phosphate free acid in high yield. The one-pot procedure is carried out in DMF or DMSO using triphenylphosphine and 2,2'-dipyridyldisulfide as the coupling agents, and 1-methylimidazole as the catalyst.

  12. Synthesis, characterizations and investigation of thermoluminescence properties of strontium pyrophosphate doped with metals

    NASA Astrophysics Data System (ADS)

    Ilkay, L. S.; Ozbayoglu, G.; Yilmaz, A.

    2014-11-01

    Strontium pyrophosphate, Sr2P2O7, was synthesized by solid-state synthesis method; the product was co-doped with copper-silver (Cu-Ag), copper-indium (Cu-In) and manganese-praseodymium (Mn-Pr) oxides (CuO, MnO, In2O3, Pr6O11 and AgNO3) by solid-state reaction method. The variation of dopant concentrations was investigated from 0.5 to 15% by weight. In addition to these processes, chemical characterizations of samples and the investigation of thermoluminescence (TLD) properties of strontium pyrophosphate with and without dopants were conducted. For the characterization; powder X-ray Diffraction (XRD) were implemented for phase purity of samples. Fourier Transform Infrared Spectroscopy (FTIR) was used to determine whether the bond structures were affected from the doping or not. Thermoluminescence (TLD) analyses were conducted on strontium pyrophosphate doped with different amounts of dopants for the first time. Glow curves showed that intensities were affected by different amounts of dopants. It can be concluded from that strontium pyrophosphate doped with 7% MnO and 1% Pr6O11 had the most powerful peak intensity around 160 °C and dosimetric property for promising application.

  13. Structure of a heterotetrameric geranyl pyrophosphate synthase from mint (Mentha piperita) reveals intersubunit regulation.

    PubMed

    Chang, Tao-Hsin; Hsieh, Fu-Lien; Ko, Tzu-Ping; Teng, Kuo-Hsun; Liang, Po-Huang; Wang, Andrew H-J

    2010-02-01

    Terpenes (isoprenoids), derived from isoprenyl pyrophosphates, are versatile natural compounds that act as metabolism mediators, plant volatiles, and ecological communicators. Divergent evolution of homomeric prenyltransferases (PTSs) has allowed PTSs to optimize their active-site pockets to achieve catalytic fidelity and diversity. Little is known about heteromeric PTSs, particularly the mechanisms regulating formation of specific products. Here, we report the crystal structure of the (LSU . SSU)(2)-type (LSU/SSU = large/small subunit) heterotetrameric geranyl pyrophosphate synthase (GPPS) from mint (Mentha piperita). The LSU and SSU of mint GPPS are responsible for catalysis and regulation, respectively, and this SSU lacks the essential catalytic amino acid residues found in LSU and other PTSs. Whereas no activity was detected for individually expressed LSU or SSU, the intact (LSU . SSU)(2) tetramer produced not only C(10)-GPP at the beginning of the reaction but also C(20)-GGPP (geranylgeranyl pyrophosphate) at longer reaction times. The activity for synthesizing C(10)-GPP and C(20)-GGPP, but not C(15)-farnesyl pyrophosphate, reflects a conserved active-site structure of the LSU and the closely related mustard (Sinapis alba) homodimeric GGPPS. Furthermore, using a genetic complementation system, we showed that no C(20)-GGPP is produced by the mint GPPS in vivo. Presumably through protein-protein interactions, the SSU remodels the active-site cavity of LSU for synthesizing C(10)-GPP, the precursor of volatile C(10)-monoterpenes.

  14. Surface immobilizable chelator for label-free electrical detection of pyrophosphate.

    PubMed

    Liu, David J; Credo, Grace M; Su, Xing; Wu, Kai; Lim, Hsiao C; Elibol, Oguz H; Bashir, Rashid; Varma, Madoo

    2011-08-07

    A new pyrophosphate (PPi) chelator was designed for surface-sensitive electrical detection of biomolecular reactions. This article describes the synthesis of the PPi-selective receptor, its surface immobilization and application to label-free electrical detection on a silicon-based field-effect transistor (FET) sensor.

  15. Bioactive calcium phosphate coating formed on micro-arc oxidized magnesium by chemical deposition

    NASA Astrophysics Data System (ADS)

    Liu, G. Y.; Hu, J.; Ding, Z. K.; Wang, C.

    2011-01-01

    In order to improve the bioactivity of the micro-arc oxidized magnesium, a calcium phosphate coating was formed on the surface of micro-arc oxidized magnesium using a chemical method. The microstructures of the substrate and the calcium phosphate coating before and after the simulated body fluids (SBF) incubation were characterized by X-ray diffraction, Fourier-transformed infrared spectroscopy and scanning electron microscopy. The results showed that the calcified coating was composed of calcium deficient hydroxyapatite (HA) and dicalcium phosphate dihydrate (DCPD). After SBF incubation, some new apatite formation on the calcified coating surface from SBF could be found. The corrosion behaviours of the samples in SBF were also investigated by potentiodynamic polarization curves and immersion tests. The results showed that calcium phosphate coating increased the corrosion potential, and decreased the hydrogen gas release.

  16. Molecular mechanisms of crystallization impacting calcium phosphate cements

    PubMed Central

    Giocondi, Jennifer L.; El-Dasher, Bassem S.; Nancollas, George H.; Orme, Christine A.

    2010-01-01

    The biomineral calcium hydrogen phosphate dihydrate (CaHPO4·2H2O), known as brushite, is a malleable material that both grows and dissolves faster than most other calcium minerals, including other calcium phosphate phases, calcium carbonates and calcium oxalates. Within the body, this ready formation and dissolution can play a role in certain diseases, such as kidney stone and plaque formation. However, these same properties, along with brushite’s excellent biocompatibility, can be used to great benefit in making resorbable biomedical cements. To optimize cements, additives are commonly used to control crystallization kinetics and phase transformation. This paper describes the use of in situ scanning probe microscopy to investigate the role of several solution parameters and additives in brushite atomic step motion. Surprisingly, this work demonstrates that the activation barrier for phosphate (rather than calcium) incorporation limits growth kinetics and that additives such as magnesium, citrate and bisphosphonates each influence step motion in distinctly different ways. Our findings provide details of how, and where, molecules inhibit or accelerate kinetics. These insights have the potential to aid in designing molecules to target specific steps and to guide synergistic combinations of additives. PMID:20308110

  17. Effect of tetrasodium pyrophosphate on the physicochemical properties of yogurt gels.

    PubMed

    Ozcan, T; Lucey, J A; Horne, D S

    2008-12-01

    The effect of tetrasodium pyrophosphate (TSPP) on the properties of yogurt gels was investigated. Various concentrations (0.05 to 0.2%) of TSPP were added to preheated (85 degrees C for 30 min) reconstituted skim milk, which was readjusted to pH 6.50. Milk was inoculated with 2% starter culture and incubated at 42 degrees C until the pH reached 4.6. Acid-base buffering profiles of milk and total and soluble calcium levels were measured. Turbidity measurements were used to indicate changes in casein dispersion. Storage modulus (G') and loss tangent (LT) values of yogurts were monitored during fermentation using dynamic oscillatory rheology. Large deformation properties of gels were also measured. Microstructural properties of yogurt were observed using fluorescence microscopy. The addition of TSPP resulted in the disappearance of the buffering peak during acid titration at pH approximately 5.1 that is due to the solubilization of colloidal calcium phosphate (CCP), and a new peak was observed at lower pH values (pH 4.0-4.5). The buffering peak at pH 6.0 during base titration virtually disappeared with addition of TSPP and a new peak appeared at pH approximately 4.8. The addition of TSPP reduced the soluble Ca content of milk and increased casein-bound Ca values. The addition of up to 0.125% TSPP resulted in a reduction in turbidity because of micelle dispersion but at 0.15%, turbidity increased and these samples exhibited a time-dependent increase in turbidity because of aggregation of casein particles. Gels made with 0.20% TSPP were very weak and had a very high gelation pH (6.35), probably due to complete dispersion of the micelle structure in this sample. The LT value of gels at pH 5.1 decreased with an increase in TSPP concentration, probably due to the loss of CCP with the addition of TSPP. The G' values at pH 4.6 of gels made with or=0.125% TSPP significantly decreased G' values. The addition

  18. Apatite coating of electrospun PLGA fibers using a PVA vehicle system carrying calcium ions.

    PubMed

    Kim, In Ae; Rhee, Sang-Hoon

    2010-01-01

    A novel method to coat electrospun poly(D,L-lactic-co-glycolic acid) (PLGA) fiber surfaces evenly and efficiently with low-crystalline carbonate apatite crystals using a poly(vinyl alcohol) (PVA) vehicle system carrying calcium ions was presented. A non-woven PLGA fabric was prepared by electrospinning: a 10 wt% PLGA solution was prepared using 1,1,3,3-hexafluoro-2-propanol as a solvent and electrospun under a electrical field of 1 kV/cm using a syringe pump with a flowing rate of 3 ml/h. The non-woven PLGA fabric, 12 mm in diameter and 1 mm in thickness, was cut and then coated with a PVA solution containing calcium chloride dihydrate (specimen PPC). As controls, pure non-woven PLGA fabric (specimen P) and fabric coated with a calcium chloride dihydrate solution without PVA (specimen PC) were also prepared. Three specimens were exposed to simulated body fluid for 1 week and this exposure led to form uniform and complete apatite coating layer on the fiber surfaces of specimen PPC. However, no apatite had formed to the fiber surfaces of specimen P and only inhomogeneous coating occurred on the fiber surfaces of specimen PC. These results were explained in terms of the calcium chelating and adhesive properties of PVA vehicle system. The practical implication of the results is that this method provides a simple but efficient technique for coating the fiber surface of an initially non-bioactive material with low-crystalline carbonate apatite.

  19. Catalytic effects of ZnO nanorods grown by sonochemical decomposition of zinc acetate dihydrate.

    PubMed

    Cho, Seok Cheol; Lee, Ho Suk; Sohn, Sang Ho

    2012-07-01

    In this study, we prepared ZnO nanorods by a sonochemical method using a zinc acetate dihydrate as a new precursor. Well-aligned high-quality ZnO nanorods were synthesized on FTO glass by the sonochemical decomposition of zinc acetate dihydrate using a ZnO thin-film as the catalytic layer. The ZnO thin-films were deposited on the FTO glass by a sputtering method. To investigate their catalytic effects on the ZnO nanorods, catalytic ZnO thin-films of 20 nm, 40 nm, and 60 nm thickness were prepared by adjusting the sputtering time. The ZnO nanorods grown on catalytic layers with different thicknesses were characterized by SEM, XRD, and PL. The ZnO nanorods grown on the catalytic layer of 40 nm thickness show the best crystal and spatial orientation and as a result display the best optical properties. It was found that a catalytic ZnO thin-film of 40 nm in thickness yields well-aligned high-quality ZnO nanorods, due to its small surface roughness and structural strain.

  20. In situ dehydration of carbamazepine dihydrate: a novel technique to prepare amorphous anhydrous carbamazepine.

    PubMed

    Li, Y; Han, J; Zhang, G G; Grant, D J; Suryanarayanan, R

    2000-01-01

    The purposes of this project were to prepare amorphous carbamazepine by dehydration of crystalline carbamazepine dihydrate, and to study the kinetics of crystallization of the prepared amorphous phase. Amorphous carbamazepine was formed and characterized in situ in the sample chamber of a differential scanning calorimeter (DSC), a thermogravimetric analyzer (TGA), and a variable temperature x-ray powder diffractometer (VTXRD). It has a glass transition temperature of 56 degrees C and it is a relatively strong glass with a strength parameter of 37. The kinetics of its crystallization were followed by isothermal XRD, under a controlled water vapor pressure of 23 Torr. The crystallization kinetics are best described by the three-dimensional nuclear growth model with rate constants of 0.014, 0.021, and 0.032 min-1 at 45, 50, and 55 degrees C, respectively. When the Arrhenius equation was used, the activation energy of crystallization was calculated to be 74 kJ/mol in the presence of water vapor (23 Torr). On the basis of the Kissinger plot, the activation energy of crystallization in the absence of water vapor (0 Torr water vapor pressure) was determined to be 157 kJ/mol. Dehydration of the dihydrate is a novel method to prepare amorphous carbamazepine; in comparison with other methods, it is a relatively gentle and effective technique.

  1. Polymer-based material containing calcium phosphate particles functionalized with a dimethacrylate monomer for use in restorative dentistry.

    PubMed

    Rodrigues, Marcela C; Xavier, Tathy A; Arana-Chavez, Victor E; Braga, Roberto R

    2016-11-23

    Dicalcium phosphate dihydrate particles functionalized with triethyleneglycol dimethacrylate were synthesized and added to a photocurable mixture of bisphenol-A glycidyl dimethacrylate and triethyleneglycol dimethacrylate with the purpose of developing a resin composite capable of releasing calcium and phosphate ions to foster dental remineralization. Particle functionalization would minimize the deleterious effect of adding low cohesive strength nano-structured particles with no chemical interaction with the organic matrix on the material's mechanical properties. The results showed that calcium release over 28 days was not impaired by particle functionalization. A statistically significant 32% increase in strength was recorded with the use of functionalized dicalcium phosphate dihydrate in comparison to the material containing non-functionalized particles. However, the strength of the unfilled resin was not matched by the composite with functionalized particles. Elastic modulus increased with particle incorporation, regardless of functionalization. Degree of conversion and optical properties (total transmittance and color change/ΔE) of the resin-based materials were not affected by the addition of dicalcium phosphate dihydrate particles (functionalized or not).

  2. A Neat Trick Using Oxalic Acid Dihydrate and Potassium Permanganate and Other Experiments with Small Organic Amine or Oxygenated Compounds

    ERIC Educational Resources Information Center

    Kelland, Malcolm A.

    2011-01-01

    Solid potassium permanganate (KMnO[subscript 4]) is shown to react in a variety of ways with small organic amines or oxygenated compounds depending on whether they are liquids or solids and whether water is present. In particular, its reaction with solid oxalic acid dihydrate can be initiated by the moisture in one's breath, making an intriguing…

  3. A Neat Trick Using Oxalic Acid Dihydrate and Potassium Permanganate and Other Experiments with Small Organic Amine or Oxygenated Compounds

    ERIC Educational Resources Information Center

    Kelland, Malcolm A.

    2011-01-01

    Solid potassium permanganate (KMnO[subscript 4]) is shown to react in a variety of ways with small organic amines or oxygenated compounds depending on whether they are liquids or solids and whether water is present. In particular, its reaction with solid oxalic acid dihydrate can be initiated by the moisture in one's breath, making an intriguing…

  4. Higher plasma quercetin levels following oral administration of an onion skin extract compared with pure quercetin dihydrate in humans.

    PubMed

    Burak, Constanze; Brüll, Verena; Langguth, Peter; Zimmermann, Benno F; Stoffel-Wagner, Birgit; Sausen, Udo; Stehle, Peter; Wolffram, Siegfried; Egert, Sarah

    2017-02-01

    To investigate the plasma kinetics of quercetin derived from hard capsules filled with onion skin extract powder or quercetin dihydrate in humans. In a randomized, single-blind, diet-controlled crossover study, 12 healthy subjects (six men and six women) aged 21-33 years were administered a single oral supra-nutritional dose of approximately 163 mg quercetin derived from onion skin extract powder (containing 95.3 % of total flavonoids as quercetin aglycone) or quercetin dihydrate (134 mg quercetin aglycone equivalent). Blood samples were collected before and during a 24-h period after quercetin administration. The concentrations of quercetin and its two monomethylated derivatives, isorhamnetin (3'-O-methyl quercetin), and tamarixetin (4'-O-methyl quercetin), were measured using HPLC with fluorescence detection after plasma enzymatic treatment. The systemic availability, determined by comparing the plasma concentration-time curves of quercetin, was 4.8 times higher, and the maximum plasma concentration (C max) was 5.4 times higher after ingestion of the onion skin extract than after ingestion of pure quercetin dihydrate. By contrast, t max did not differ significantly between the two formulations. The C max values for isorhamnetin and tamarixetin were 3.8 and 4.4 times higher, respectively, after administration of onion skin extract than after pure quercetin dihydrate. The plasma kinetics of quercetin were not significantly different in men and women. Quercetin aglycone derived from onion skin extract powder is significantly more bioavailable than that from quercetin dihydrate powder filled hard capsules.

  5. Formation of pyrophosphate, tripolyphosphate, and phosphorylimidazole with the thioester, N, S-diacetylcysteamine, as the condensing agent. [molecular evolution

    NASA Technical Reports Server (NTRS)

    Weber, A. L.

    1981-01-01

    The formation of pyrophosphate, tripolyphosphate and phosphorylimidazole from orthophosphate in the presence of the thioester N, S-diacetylcysteamine is reported. Reactions performed with 0.20 M orthophosphate and 0.20 M N, S-diacetylcysteamine in 0.40 M imidazole under drying conditions at 50 C yielded pyrophosphate and tripolyphosphate after six days in the presence and absence of divalent metal ions. Reactions carried out at ambient temperature yielded phosphorylimidazole in the presence or absence of 0.05 M MgCl2, phosphorylimidazole and pyrophosphate in the presence of 0.05 M CaCl2, and pyrophosphate and tripolyphosphate in the presence of 0.15 M CaCl2. Such reactions represent potential pathways for the formation of energy-rich compounds providing free energy for use in prebiotic biopolymer synthesis.

  6. Formation of pyrophosphate, tripolyphosphate, and phosphorylimidazole with the thioester, N, S-diacetylcysteamine, as the condensing agent. [molecular evolution

    NASA Technical Reports Server (NTRS)

    Weber, A. L.

    1981-01-01

    The formation of pyrophosphate, tripolyphosphate and phosphorylimidazole from orthophosphate in the presence of the thioester N, S-diacetylcysteamine is reported. Reactions performed with 0.20 M orthophosphate and 0.20 M N, S-diacetylcysteamine in 0.40 M imidazole under drying conditions at 50 C yielded pyrophosphate and tripolyphosphate after six days in the presence and absence of divalent metal ions. Reactions carried out at ambient temperature yielded phosphorylimidazole in the presence or absence of 0.05 M MgCl2, phosphorylimidazole and pyrophosphate in the presence of 0.05 M CaCl2, and pyrophosphate and tripolyphosphate in the presence of 0.15 M CaCl2. Such reactions represent potential pathways for the formation of energy-rich compounds providing free energy for use in prebiotic biopolymer synthesis.

  7. Stimulation of bovine heart pyruvate dehydrogenase kinase by. cap alpha. -ketoisovalerate in the presence of thiamin pyrophosphate

    SciTech Connect

    Robertson, J.G.; Barron, L.L.; Olson, M.S.

    1987-05-01

    Purified bovine heart pyruvate dehydrogenase complex was used to study pyruvate dehydrogenase kinase (PDH kinase) regulation. Previously, they showed that KCl and NH4Cl stimulate PDH kinase, and that thiamin pyrophosphate over the range of 1-80 ..mu..M completely blocks PDH kinase stimulation by 20 mM KCl, whereas thiamin pyrophosphate has very little inhibitory effect in the absence of KCl. Under inhibitory conditions, 100 ..mu..M thiamin pyrophosphate and 20 mM KCl, addition of 1 mM ..cap alpha..-ketoisovalerate stimulated PDH kinase activity 1.5-fold. Addition of 1 mM ..cap alpha..-ketoisovalerate had a similar stimulatory effect in the presence of 1 mM NH4Cl and 100 ..mu..M thiamin pyrophosphate. Half-maximal stimulation occurred at about 200 ..mu..M ..cap alpha..-ketoisovalerate in both cases. Inhibition by thiamin pyrophosphate was uncompetitive, and the effect of ..cap alpha..-ketoisovalerate on thiamin pyrophosphate inhibition was to shift both Vm and Km toward control values without changing the slope of the reciprocal plot. After incubation of pyruvate dehydrogenase complex with ..cap alpha..-ketoisovalerate and gel filtration on Sephadex G-25 to remove ..cap alpha..-ketoisovalerate, PDH kinase activity remained stimulated relative to controls incubated with only buffer. In bovine heart mitochondria solubilized in 0.2% Triton X-100 and incubated with (..gamma..-/sup 32/P)ATP, ..cap alpha..-ketoisovalerate stimulated PDH phosphorylation in the presence of thiamin pyrophosphate, as detected by SDS-PAGE and autoradiography. The results indicate that acylation of pyruvate dehydrogenase complex results in PDH kinase stimulation, but only in the presence of monovalent cation and thiamin pyrophosphate. Furthermore, the results also suggest that this effect may be relevant under more in vivo conditions.

  8. Modulation of polyepoxysuccinic acid on crystallization of calcium oxalate

    SciTech Connect

    Zhang, Yanqing; Tang, Yongming; Xu, Jinqiu; Zhang, Dongqin; Lu, Gang; Jing, Wenheng

    2015-11-15

    The influence of polyepoxysuccinic acid (PESA) on the phase composition and crystal morphology of calcium oxalate was investigated in this paper. It was found that the presence of PESA inhibited the growth of the monoclinic calcium oxalate monohydrate (COM) crystal and promoted the nucleation of the tetragonal calcium oxalate dihydrate (COD). In addition, with the increase in PESA concentration, the aggregation of COD crystals was reduced but the particle size was increased. Under the conditions of low calcium-to-oxalate ratio and high CaOx concentration, PESA could not effectively stabilize the formation of COD. Based on molecular dynamic simulations, the adsorption of PESA on CaOx crystal faces was confirmed. - Graphical abstract: Introduction of PESA into crystallization solutions promotes the formation of calcium oxalate dehydrate and modifies the morphology of crystals. - Highlights: • PESA induces the formation of COD at low supersaturation. • Establishment of Ca-rich surface augments the adsorption of PESA. • At Ca/Ox=0.5 PESA cannot induce the formation of COD compared with Ca/Ox=2. • Interaction of PESA with COM faces is stronger than that with COD faces.

  9. Experimental models of renal calcium stones in rodents

    PubMed Central

    Bilbault, Héloïse; Haymann, Jean-Philippe

    2016-01-01

    In human nephrolithiasis, most stones are containing calcium and are located within urinary cavities; they may contain monohydrate calcium oxalate, dihydrate calcium oxalate and/or calcium phosphates in various proportion. Nephrolithiasis may also be associated with nephrocalcinosis, i.e., crystal depositions in tubular lumen and/or interstitium, an entity which suggests specific pathological processes. Several rodents models have been developed in order to study the pathophysiology of intrarenal crystal formation. We review here calcium rodent models classified upon the presence of nephrolithiasis and/or nephrocalcinosis. As rodents are not prone to nephrolithiasis, models require the induction of a long standing hypercalciuria or hyperoxaluria (thus explaining the very few studies reported), conversely to nephrocalcinosis which may occur within hours or days. Whereas a nephrotoxicity leading to tubular injury and regeneration appears as a critical event for crystal retention in nephrocalcinosis models, surprisingly very little is known about the physiopathology of crystal attachment to urothelium in nephrolithiasis. Creating new models of nephrolithiasis especially in different genetic mice strains appears an important challenge in order to unravel the early mechanisms of urinary stone formation in papilla and fornices. PMID:26981444

  10. Thiamine pyrophosphate (TPP) negatively regulates transcription of some thi genes of Salmonella typhimurium.

    PubMed

    Webb, E; Febres, F; Downs, D M

    1996-05-01

    In Salmonella typhimurium, thiamine is a required nutrient that is synthesized de novo. Labeling studies have demonstrated probable precursors for both the 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate moiety and the 4-methyl-5-(beta-hydroxyethyl) thiazole monophosphate moiety. The isolation of thiamine auxotrophs with mutations in at least five different genetic loci is reported. The majority (22 of 25) of the mutants required only the thiazole moiety of thiamine to satisfy their growth requirement. Most (14 of 25) of the mutants were affected in the thi cluster at min 90 on the S. typhimurium genetic map. Data provided herein indicate that this cluster encodes an operon whose transcription is regulated by thiamine and suggest that thiamine pyrophosphate, or a molecule derived form it, is the effector molecule. Mutants with altered regulation of this operon were isolated, and we propose that they are defective in thiamine phosphate kinase, the product of the thiL gene.

  11. Muscle necrosis in the extremities: evaluation with Tc-99m pyrophosphate scanning--a retrospective review

    SciTech Connect

    Timmons, J.H.; Hartshorne, M.F.; Peters, V.J.; Cawthon, M.A.; Bauman, J.M.

    1988-04-01

    A retrospective review was done of 34 extremities studied between 1981 and 1985 with technetium-99m pyrophosphate scanning; 22 were subsequently amputated. Results of detailed pathologic examination or immediate postoperative examination of the resected extremity were available in 16 cases. In these cases, scanning had allowed correct prediction of the level of amputation and of the specific areas of muscle infarction in 13 cases. In the one case in which amputation was performed for infection rather than muscle necrosis, the lack of necrosis was correctly predicted with the scan. The limited results of this study indicate that the Tc-99m pyrophosphate scan allows the location of necrotic muscle to be predicted accurately and may therefore be a useful adjunct in determining the best level for ultimate amputation. Special caution is required in those cases in which muscle necrosis is due to acute causes (e.g., traumatic thrombosis) rather than chronic vascular disease.

  12. Pyrophosphate selective fluorescent chemosensors: cascade recognition of nuclear stain mimicking DAPI.

    PubMed

    Goswami, Shyamaprosad; Das, Avijit Kumar; Pakhira, Bholanath; Basu Roy, Sohini; Maity, Anup Kumar; Saha, Partha; Sarkar, Sabyasachi

    2014-09-07

    A new zinc(ii) complex with a condensed hydroxynaphthyl pyridine (SPHN) as the coordinated ligand has been synthesized for the selective recognition of pyrophosphate (PPi) over other anions including phosphate in a mixed aqueous solution. The fluorescence enhancement of SPHN in association with Zn(2+) ions is quenched in the presence of intracellular pyrophosphate. This phenomenon is utilized in the construction of a logic gate. The binding of SPHN with Zn(2+) and its displacement by PPi have been established by photophysical investigation and supported by the DFT level of studies. The development of blue fluorescence in the {} complex upon binding of zinc with is shown to be useful as a nucleus marker in a cell similar to the commercially available staining compound, DAPI (diamino-2-phenylindole).

  13. Thiamine pyrophosphate stimulates acetone activation by Desulfococcus biacutus as monitored by a fluorogenic ATP analogue.

    PubMed

    Gutiérrez Acosta, Olga B; Hardt, Norman; Hacker, Stephan M; Strittmatter, Tobias; Schink, Bernhard; Marx, Andreas

    2014-06-20

    Acetone can be degraded by aerobic and anaerobic microorganisms. Studies with the strictly anaerobic sulfate-reducing bacterium Desulfococcus biacutus indicate that acetone degradation by these bacteria starts with an ATP-dependent carbonylation reaction leading to acetoacetaldehyde as the first reaction product. The reaction represents the second example of a carbonylation reaction in the biochemistry of strictly anaerobic bacteria, but the exact mechanism and dependence on cofactors are still unclear. Here, we use a novel fluorogenic ATP analogue to investigate its mechanism. We find that thiamine pyrophosphate is a cofactor of this ATP-dependent reaction. The products of ATP cleavage are AMP and pyrophosphate, providing first insights into the reaction mechanism by indicating that the reaction proceeds without intermediate formation of acetone enol phosphate.

  14. Repeptization by dissolution in a colloidal system of iron(III) pyrophosphate.

    PubMed

    van Leeuwen, Y Mikal; Velikov, Krassimir P; Kegel, Willem K

    2012-12-04

    Repeptization (redispersion) from an aggregated state is usually only possible in charge-stabilized colloidal systems if the system is either coagulated in the secondary minimum of the interaction potential or if the system cannot settle completely into the primary minimum. In this work, we analyze the zeta potential, conductivity, and long-term stability of colloidal systems of iron(III) pyrophosphate and surprisingly find that the system seems to defy conventional wisdom as it can be repeptized from its coagulated state regardless of aging time and background ions. Moreover, after having been stored for up to a month in 2 M NaCl, dialysis of iron pyrophosphate will yield a colloidal dispersion that is actually stable for a longer period of time than a fresh system with background electrolyte removed.

  15. Technetium-99m stannous pyrophosphate scintigraphy in patients with calcification within the cardiac silhouette.

    PubMed Central

    Wald, R W; Sternberg, L; Huckell, V F; Staniloff, H M; Feiglin, D H; Morch, J E

    1978-01-01

    Technetium-99m stannous pyrophosphate scintiscanning was performed in 22 patients with radiographically detected calcification within the cardiac silhouette. All but one of these scintigrams showed a localised area of increased activity similar to that ordinarily seen in acute myocardial infarction. Scintiscans in 3 patients after removal of the calcified aortic valve reverted to negative. It was concluded that this technique for acute infarct detection may yield false positive results in the presence of cardiac calcification. Images PMID:207292

  16. Technetium-99m stannous pyrophosphate scintigraphy in patients with calcification within the cardiac silhouette.

    PubMed

    Wald, R W; Sternberg, L; Huckell, V F; Staniloff, H M; Feiglin, D H; Morch, J E

    1978-05-01

    Technetium-99m stannous pyrophosphate scintiscanning was performed in 22 patients with radiographically detected calcification within the cardiac silhouette. All but one of these scintigrams showed a localised area of increased activity similar to that ordinarily seen in acute myocardial infarction. Scintiscans in 3 patients after removal of the calcified aortic valve reverted to negative. It was concluded that this technique for acute infarct detection may yield false positive results in the presence of cardiac calcification.

  17. Imaging necrotic myocardium: Detection with 99mTc-pyrophosphate and radiolabeled antimyosin

    SciTech Connect

    Khaw, B.A.; Haber, E. )

    1989-08-01

    The major value of hot-spot imaging of the myocardium is its ability to define areas of necrosis rather than areas of diminished blood flow or cellular function. Applications of hot-spot imaging include the diagnosis and quantitation of myocardial infarction, myocarditis, and cardiac transplant rejection. The two agents in clinical use, 99mTc-Pyrophosphate and radiolabeled antimyosin, are discussed. 52 references.

  18. Crystal growth and characterization of sodium p-nitrophenolate dihydrate (NPNa) single crystals for NLO applications

    NASA Astrophysics Data System (ADS)

    Sethupathi, D.; Pandian, Muthu Senthil; Maurya, K. K.; Ramasamy, P.

    2017-05-01

    A good quality and transparent semi-organic nonlinear optical sodium p-nitrophenolate dihydrate (NPNa) crystal has been grown by slow evaporation solution technique (SEST) in a period of 180 days. The single crystal XRD confirms the NPNa crystal belongs to orthorhombic system with the non-centrosymmetry space group Ima2. The quality of the grown crystal was examined by high resolution X-ray diffraction (HXRD) analysis and the full width at half maximum (FWHM) of NPNa crystal was found to be 19 arc sec. The thermal property of the NPNa crystal was analyzed by TG-DTA analysis. The second harmonic generation (SHG) efficiency of the grown crystal was studied by Kurtz-Perry powder technique and the calculated SHG conversion efficiency was found to be 2.4 times that of standard KDP material.

  19. Growth, photoluminescence, thermal and mechanical behaviour of Ethyltriphenylphosphonium bromide dihydrate crystal

    NASA Astrophysics Data System (ADS)

    Parthasarathy, M.; Gopalakrishnan, R.

    2013-10-01

    Single crystal of Ethyltriphenylphosphonium bromide dihydrate (ETPB) was grown by slow evaporation solution growth technique. The grown crystal was confirmed by single crystal X-ray diffraction. The functional groups and vibrational frequencies were identified using FT-IR and FT-Raman spectral analyses. Optical properties were studied by UV-Visible and photoluminescence spectroscopic techniques to explore its efficacy towards device fabrication. Thermal characteristics of ETPB were studied using the TGA/DTA and DSC response curves. The mechanical behaviour of the grown crystal was studied using Vicker's microhardness tester and the work hardening coefficient was evaluated. The second harmonic generation of the title compound was tested by Kurtz-Perry powder technique.

  20. The EFG tensors and the positions of the deuterons in deuterated pyromellitic acid dihydrate

    NASA Astrophysics Data System (ADS)

    Schajor, W.; Tegenfeldt, J.; Haeberlen, U.

    We report on high-field FT NMR measurements of the EFG tensors at the sites of the deuterons in selectively deuterated single crystals of pyromellitic acid dihydrate, PMADH. The principal directions of the EFGs and the quadrupole coupling constants are used to derive information about the positions of the deuterons relative to the framework of the heavy atoms of the PMA molecule, and about the hydrogen bond network in PMADH. The results are compared with the hydrogen positions found in Takusagawa's X-ray study of PMADH. The D NMR and the X-ray results are at variance. It is concluded that at least in this case of PMADH D NMR is superior to X-ray diffraction with regard to the positioning of the hydrogens.

  1. Crystal structure of 3,5-di­methyl­pyridine N-oxide dihydrate

    PubMed Central

    Merino García, Rosario; Ríos-Merino, Francisco Javier; Bernès, Sylvain; Reyes-Ortega, Yasmi

    2016-01-01

    In the title compound, also known as 3,5-lutidine N-oxide dihydrate, C7H9NO·2H2O, the N—O bond is weakened due to the involvement of the O atom as an acceptor of hydrogen bonds from the two water mol­ecules of crystallization present in the asymmetric unit. Fused R 3 5(10) ring motifs based on O—H⋯O hydrogen bonds form chains in the [010] direction, which are further connected by weak C—H⋯O inter­molecular contacts. As a result, the lutidine mol­ecules are stacked in an efficient manner, with π–π contacts characterized by a short separation of 3.569 (1) Å between the benzene rings. PMID:27980810

  2. Mapping of the pyrophosphate binding sites of beef heart mitochondrial F1-ATPase by photolabelling with azidonitrophenyl [alpha-32P]pyrophosphate.

    PubMed

    Michel, L; Garin, J; Vinçon, M; Gagnon, J; Vignais, P

    1995-02-14

    4-Azido-2-nitrophenyl [alpha-32P]pyrophosphate (azido-[alpha-32P]PPi) mimics ADP and PPi by some of its binding properties when assayed in the absence of photoirradiation with mitochondrial F1-ATPase. Upon photoirradiation, both alpha- and beta-subunits of F1-ATPase were covalently labelled. Following chemical and enzymatic cleavages of each of the two photolabelled subunits, peptides containing the covalently bound radioactivity were separated by HPLC and identified by amino acid sequencing. Bound azido-[alpha-32P]PPi was found to be concentrated in two distant sequences of the alpha-subunit, namely Asp194-Thr221 and Lys386-Met437, and in a single sequence of the beta-subunit Glu294-Met358 with most of the photoprobe bound to beta-Tyr-311 and beta-Tyr-345. These results are discussed in terms of a model in which the pyrophosphate binding sites of F1 are located in regions of the alpha- and beta-subunits exposed at the interface between the two subunits and correspond to non-catalytic and catalytic adenine nucleotide binding sites, respectively.

  3. Simple tools for assembling and searching high-density picolitre pyrophosphate sequence data.

    PubMed

    Parker, Nicolas J; Parker, Andrew G

    2008-04-18

    The advent of pyrophosphate sequencing makes large volumes of sequencing data available at a lower cost than previously possible. However, the short read lengths are difficult to assemble and the large dataset is difficult to handle. During the sequencing of a virus from the tsetse fly, Glossina pallidipes, we found the need for tools to search quickly a set of reads for near exact text matches. A set of tools is provided to search a large data set of pyrophosphate sequence reads under a "live" CD version of Linux on a standard PC that can be used by anyone without prior knowledge of Linux and without having to install a Linux setup on the computer. The tools permit short lengths of de novo assembly, checking of existing assembled sequences, selection and display of reads from the data set and gathering counts of sequences in the reads. Demonstrations are given of the use of the tools to help with checking an assembly against the fragment data set; investigating homopolymer lengths, repeat regions and polymorphisms; and resolving inserted bases caused by incomplete chain extension. The additional information contained in a pyrophosphate sequencing data set beyond a basic assembly is difficult to access due to a lack of tools. The set of simple tools presented here would allow anyone with basic computer skills and a standard PC to access this information.

  4. Substrate specificity and membrane topology of Escherichia coli PgpB, an undecaprenyl pyrophosphate phosphatase.

    PubMed

    Touzé, Thierry; Blanot, Didier; Mengin-Lecreulx, Dominique

    2008-06-13

    The synthesis of the lipid carrier undecaprenyl phosphate (C(55)-P) requires the dephosphorylation of its precursor, undecaprenyl pyrophosphate (C(55)-PP). The latter lipid is synthesized de novo in the cytosol and is also regenerated after its release from the C(55)-PP-linked glycans in the periplasm. In Escherichia coli the dephosphorylation of C(55)-PP was shown to involve four integral membrane proteins, BacA, and three members of the type 2 phosphatidic acid phosphatase family, PgpB, YbjG, and YeiU. Here, the PgpB protein was purified to homogeneity, and its phosphatase activity was examined. This enzyme was shown to catalyze the dephosphorylation of C(55)-PP with a relatively low efficiency compared with diacylglycerol pyrophosphate and farnesyl pyrophosphate (C(15)-PP) lipid substrates. However, the in vitro C(55)-PP phosphatase activity of PgpB was specifically enhanced by different phospholipids. We hypothesize that the phospholipids are important determinants to ensure proper conformation of the atypical long axis C(55) carrier lipid in membranes. Furthermore, a topological analysis demonstrated that PgpB contains six transmembrane segments, a large periplasmic loop, and the type 2 phosphatidic acid phosphatase signature residues at a periplasmic location.

  5. [Efficacy of thiamine pyrophosphate or carboxylase in the salvage of diabetic foot].

    PubMed

    Carmona-Cervantes, J

    2014-01-01

    Diabetic foot represents one of the most common complications in patients with a long standing disease. The etiology is neuropathy, infections and ischemia that together contribute to the sequence of tissue necrosis, ulceration and gangrene. Since treatment is very difficult, we must look for several options to solve these problems caused by chronic hyperglycemia. Thiamine pyrophosphate or carboxylase perform multiple metabolic and non-metabolic activities that are considered important in the resolution of diabetic impairments, therefore, this work shows the results when using it in patients with diabetic foot. 29 patients with diabetic foot were treated between January 1998 and July 2012: 19 Wagner type III and 12 Wagner type IV. Management was the administration of antibiotics, partial surgical procedures and thiamine pyrophosphate. The infectious process was controlled, the appearance of granulation tissue and scarring of the lesion in a period of 2 to 6 months depending on the severity of the problem. Given the clinical data and evolution of the patients, we conclude that the administration of thiamine pyrophosphate was able to control metabolic and non-metabolic dysfunctions that lead to complications in diabetic patients, therefore we must consider it a tool in the treatment of diabetic patients in general and for diabetic foot salvage in particular.

  6. Structural basis of thiamine pyrophosphate analogues binding to the eukaryotic riboswitch.

    PubMed

    Thore, Stéphane; Frick, Christian; Ban, Nenad

    2008-07-02

    The thiamine pyrophosphate (TPP)-sensing riboswitch is the only riboswitch found in eukaryotes. In plants, TPP regulates its own production by binding to the 3' untranslated region of the mRNA encoding ThiC, a critical enzyme in thiamine biosynthesis, which promotes the formation of an unstable splicing variant. In order to better understand the molecular basis of TPP-analogue binding to the eukaryotic TPP-responsive riboswitch, we have determined the crystal structures of the Arabidopsis thaliana TPP-riboswitch in complex with oxythiamine pyrophosphate (OTPP) and with the antimicrobial compound pyrithiamine pyrophosphate (PTPP). The OTPP-riboswitch complex reveals that the pyrimidine ring of OTPP is stabilized in its enol form in order to retain key interactions with guanosine 28 of the riboswitch previously observed in the TPP complex. The structure of PTPP in complex with the riboswitch shows that the base moiety of guanosine 60 undergoes a conformational change to cradle the pyridine ring of the PTPP. Structural information from these complexes has implications for the design of novel antimicrobials targeting TPP-sensing riboswitches.

  7. Regularities of extracting humic acids from soils using sodium pyrophosphate solutions

    NASA Astrophysics Data System (ADS)

    Bakina, L. G.; Drichko, V. F.; Orlova, N. E.

    2017-02-01

    Regularities of extracting humic acids from different soil types (soddy-podzolic soil, gray forest soil, and all chernozem subtypes) with sodium pyrophosphate solutions at different pH values (from 5 to 13) have been studied. It is found that, regardless of soil type, the process occurs in two stages through the dissociation of carboxylic groups and phenolic hydroxyls, each of which can be described by a logistic function. Parameters of the logistic equations approximating the extraction of humic acids from soils at different pH values are independent of the content and composition of humus in soils. Changes in the optical density of humic acids extracted from soils using sodium pyrophosphate solutions with different pH values are described in the first approximation by the Gaussian function. The optically densest humic acids are extracted using sodium pyrophosphate solutions at pH 10. Therefore, it is proposed to use an extract with pH 10 for the characterization of organic matter with the maximum possible degree of humification in the given soil.

  8. Preparation of cubic niobium pyrophosphate containing Nb(IV) and topatactic extraction of phosphorus atoms

    SciTech Connect

    Fukuoka, Hiroshi; Imoto, Hideo; Saito, Taro

    1995-10-01

    A reduced phase of niobium pyrophosphate containing Nb{sup 4+} has been prepared from the reaction of Nb{sup 6}Cl{sub 14}{center_dot}8H{sub 2}O and phosphoric acid. The X-ray powder diffraction and electron diffraction studies have shown that the compound belongs to the Pa3 space group and has the ZrP{sub 2}O{sub 7} structure with a cubic superstructure (a{prime} = 3a{sub 0}). Magnetic susceptibility was measured for two samples, and the mean oxidation numbers of niobium in them are deduced to by + 4.66 and +4.88. The cell constants of these samples are a = 8.0830(4) and 8.0705(2) {angstrom}, respectively. As the mean oxidation number of niobium increases, the color of the compound varies from brown to gray. When the compound is heated in oxygen, it changes into the known white niobium pyrophosphate, in which all niobium is in the +5 oxidation state. Rietveld refinements indicate that niobium pyrophosphates have defects in the phosphorus sites. The topotactic extraction of the phosphorus atoms in the reaction with oxygen was confirmed by the analysis of phosphorus oxide generated during the reaction.

  9. Crystallization and preliminary X-ray diffraction study of phosphoribosyl pyrophosphate synthetase from E. Coli

    NASA Astrophysics Data System (ADS)

    Timofeev, V. I.; Abramchik, Yu. A.; Zhukhlistova, N. E.; Kuranova, I. P.

    2015-09-01

    Enzymes of the phosphoribosyl pyrophosphate synthetase family (PRPPS, EC 2.7.6.1) catalyze the formation of 5-phosphoribosyl pyrophosphate (5-PRPP) from adenosine triphosphate and ribose 5-phosphate. 5-Phosphoribosyl pyrophosphate is an important intermediate in the synthesis of purine, pyrimidine, and pyridine nucleotides, as well as of the amino acids histidine and tryptophan. The crystallization conditions for E. coli PRPPS were found by the vapor-diffusion technique and were optimized to apply the capillary counter-diffusion technique. The X-ray diffraction data set was collected from the crystals grown by the counter-diffusion technique using a synchrotron radiation source to 3.1-Å resolution. The crystals of PRPPS belong to sp. gr. P6322 and have the following unit-cell parameters: a = b = 104.44 Å, c = 124.98 Å, α = β = 90°, γ = 120°. The collected X-ray diffraction data set is suitable for the solution of the three-dimensional structure of PRPPS at 3.1-Å resolution.

  10. Crystallization and preliminary X-ray diffraction study of phosphoribosyl pyrophosphate synthetase from E. Coli

    SciTech Connect

    Timofeev, V. I. Abramchik, Yu. A. Zhukhlistova, N. E. Kuranova, I. P.

    2015-09-15

    Enzymes of the phosphoribosyl pyrophosphate synthetase family (PRPPS, EC 2.7.6.1) catalyze the formation of 5-phosphoribosyl pyrophosphate (5-PRPP) from adenosine triphosphate and ribose 5-phosphate. 5-Phosphoribosyl pyrophosphate is an important intermediate in the synthesis of purine, pyrimidine, and pyridine nucleotides, as well as of the amino acids histidine and tryptophan. The crystallization conditions for E. coli PRPPS were found by the vapor-diffusion technique and were optimized to apply the capillary counter-diffusion technique. The X-ray diffraction data set was collected from the crystals grown by the counter-diffusion technique using a synchrotron radiation source to 3.1-Å resolution. The crystals of PRPPS belong to sp. gr. P6{sub 3}22 and have the following unit-cell parameters: a = b = 104.44 Å, c = 124.98 Å, α = β = 90°, γ = 120°. The collected X-ray diffraction data set is suitable for the solution of the three-dimensional structure of PRPPS at 3.1-Å resolution.

  11. Gallium Arsenate Dihydrate under Pressure: Elastic Properties, Compression Mechanism, and Hydrogen Bonding.

    PubMed

    Spencer, Elinor C; Soghomonian, Victoria; Ross, Nancy L

    2015-08-03

    Gallium arsenate dihydrate is a member of a class of isostructural compounds, with the general formula M(3+)AsO4·2H2O (M(3+) = Fe, Al, In, or Ga), which are being considered as potential solid-state storage media for the sequestration of toxic arsenic cations. We report the first high-pressure structural analysis of a metal arsenate dihydrate, namely, GaAsO4·2H2O. This compound crystallizes in the orthorhombic space group Pbca with Z = 8. Accurate unit cell parameters as a function of pressure were obtained by high-pressure single-crystal X-ray diffraction, and a bulk modulus of 51.1(3) GPa for GaAsO4·2H2O was determined from a third-order Birch-Murnaghan equation of state fit to the P-V data. Assessment of the pressure dependencies of the unit cell lengths showed that the compressibility of the structure along the axial directions increases in the order of [010] < [100] < [001]. This order was found to correlate well with the proposed compression mechanism for GaAsO4·2H2O, which involves deformation of the internal channel void spaces of the polyhedral helices that lie parallel to the [010] direction, and increased distortion of the GaO6 octahedra. The findings of the high-pressure diffraction experiment were further supported by the results from variable-pressure Raman analysis of GaAsO4·2H2O. Moreover, we propose a revised and more complex model for the hydrogen-bonding scheme in GaAsO4·2H2O, and on the basis of this revision, we reassigned the peaks in the OH stretching regions of previously published Raman spectra of this compound.

  12. Influence of environmental conditions on the kinetics and mechanism of dehydration of carbamazepine dihydrate.

    PubMed

    Han, J; Suryanarayanan, R

    1998-11-01

    The object of this project was to study the influence of temperature and water vapor pressure on the kinetics and mechanism of dehydration of carbamazepine dihydrate and to establish the relationship between the dehydration mechanism and the solid-state of the anhydrous phase formed. Three experimental techniques were utilized to study the kinetics of dehydration of carbamazepine dihydrate (C15H12N2O.2H2O)-thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and variable temperature powder X-ray diffractometry (VTXRD). These techniques respectively provide information about the changes in weight, heat flow and solid-state (phase) during the dehydration process. The instrumental setup was modified so that simultaneous control of both the temperature and the water vapor pressure was possible. The experiments were carried out at different temperatures, ranging from 26 to 64 degrees C. In the absence of water vapor, the dehydration followed the 2-dimensional phase boundary controlled model at all the temperatures studied. In the next stage, the water vapor pressure was altered while the studies were carried out at a single temperature of 44 degrees C. The dehydration was 2-dimensional phase boundary controlled at water vapor pressures < or = 5.1 torr while the Avrami-Erofeev kinetics (3-dimensional nucleation) was followed at water vapor pressures > or = 12.0 torr. In the former case, the anhydrous phase formed was X-ray amorphous while it was the crystalline anhydrous gamma-carbamazepine in the latter. Thus a relationship between the mechanism of dehydration and the solid-state of the product phase was evident. The dehydration conditions influence not only the mechanism but also the solid-state of the anhydrous phase formed. While the techniques of TGA and DSC have found extensive use in studying dehydration reactions, VTXRD proved to be an excellent complement in characterizing the solid-states of the reactant and product phases.

  13. Calcium - ionized

    MedlinePlus

    ... 245. Read More Acute kidney failure Albumin - blood (serum) test Bone tumor Calcium blood test Hyperparathyroidism Hypoparathyroidism Malabsorption Milk-alkali syndrome Multiple myeloma Osteomalacia Paget disease of the bone Rickets Sarcoidosis Vitamin D Review ...

  14. Calcium - urine

    MedlinePlus

    ... Monitor someone who has a problem with the parathyroid gland , which helps control calcium levels in the blood ... much production of parathyroid hormone (PTH) by the parathyroid glands in the neck (hyperparathyroidism) Use of loop diuretics ...

  15. Effect of biomedical organic compounds on the setting reaction of calcium phosphates.

    PubMed

    Yu, Tao; Ye, Jiandong; Gao, Chengying; Yu, Long; Wang, Yingjun

    2010-01-01

    In the present study, the effect of biomedical organic compounds (starch, sodium alginate, chitosan and gelatin) on the hydration of calcium phosphates was studied using X-ray diffraction, infrared spectroscopy, scanning electron microscopy and XPS analysis. Amorphous calcium phosphate (ACP) was prepared by a mechanochemical route and mixed with biomedical organic compounds. A solidification reaction occurred between ACP and dicalcium phosphate dihydrate (DCPD); the hydration product was poorly crystallized hydroxyapatite (HA). During the setting reaction of ACP and DCPD, the presence of biomedical organic compounds had an effect on the hydration product: the bonding energies of the primary elements (Ca, P) in the hydration product (HA) were changed; also different hydration morphologies, self-setting properties, rheological properties and mechanical strength of the cement were obtained. This work will allow advances in the synthesis of bionic composite calcium phosphate cement (CPC).

  16. EVALUATION OF SURGICAL CAVITIES FILLED WITH THREE TYPES OF CALCIUM SULFATE

    PubMed Central

    Maeda, Sergio Toshinori; Bramante, Clovis Monteiro; Taga, Rumio; Garcia, Roberto Brandão; de Moraes, Ivaldo Gomes; Bernadineli, Norberti

    2007-01-01

    The aim of this study was to evaluate histologically, three types of calcium sulfate - Merck (Brazil), Surgiplaster (Italy) and Capset (USA) - in surgically created defects on rabbit femurs. Twenty male New Zealand rabbits were used. Two surgical bone defects (5 mm diameter x 8 mm depth) were created on each distal epiphysis using a #3 Dentoflex trephine bur. Defects were filled with: group 1 - di-hydrated calcium sulfate (Merck); group 2 - Capset (Lifecore-USA); group 3 - Surgiplaster (Classimport-Italy); group 4 – control (blood clot). The animals were sacrificed 30, 60, 90 and 180 days postoperatively. Semi-serial 6-mm-thick sections were obtained, stained with hematoxylin and eosin and examined under light microscopy. Bone defects treated with calcium sulfate exhibited new bone formation regardless of the product trademark. PMID:19089171

  17. Calcium Carbonate.

    PubMed

    Al Omari, M M H; Rashid, I S; Qinna, N A; Jaber, A M; Badwan, A A

    2016-01-01

    Calcium carbonate is a chemical compound with the formula CaCO3 formed by three main elements: carbon, oxygen, and calcium. It is a common substance found in rocks in all parts of the world (most notably as limestone), and is the main component of shells of marine organisms, snails, coal balls, pearls, and eggshells. CaCO3 exists in different polymorphs, each with specific stability that depends on a diversity of variables. © 2016 Elsevier Inc. All rights reserved.

  18. The role of hydrogen bonding in the thermal expansion and dehydration of brushite, di-calcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Schofield, P. F.; Knight, K. S.; Houwen, J. A. M. Van Der; Valsami-Jones, E.

    2004-12-01

    The unit-cell and atomic parameters of perdeuterated brushite have been extracted from Rietveld analysis of neutron powder diffraction data within the temperature range 4.2 to 470 K. The thermal expansion of brushite is anisotropic, with the largest expansion along the b axis due principally to the effect of the O(1)···D(4) and O(3)···D(2) hydrogen bonds. Expansion along the c axis, influenced by the Ow1···D(5) interwater hydrogen bond, is also large. The high temperature limits for the expansion coefficients for the unit-cell edges a, b and c are 9.7(5) × 10-6, 3.82(9) × 10-5 and 5.54(5) × 10-5 K-1, respectively, and for the cell volume it is 9.7(1) × 10-5 K-1. The β angle displays oscillatory variation, and empirical data analysis results in αβ = 1.28(3) × 10-6sin(0.0105 T) K-1, within this temperature range. The evolution of the thermal expansion tensor of brushite has been calculated between 50 ≤T≤ 400 K. At 300 K the magnitudes of the principal axes are α11 = 50(6) × 10-6 K-1, α22 = 26.7(7) × 10-6 K-1 and α33 = 7.0(5) × 10-6 K-1. The intermediate axis, α22, is parallel to b, and using IRE convention for the tensor orthonormal basis, the axes α11 and α33 have directions equal to (-0.228, 0, -0.974) and (-0.974, 0, 0.228) respectively. Under the conditions of these experiments, the onset of dehydration occurred at temperatures above 400 K. Bond valence analysis combined with assessments of the thermal evolution of the bonding within brushite suggests that dehydration is precipitated through instabilities in the chemical environment of the second water molecule.

  19. Calcium orthophosphates

    PubMed Central

    Dorozhkin, Sergey V.

    2011-01-01

    The present overview is intended to point the readers’ attention to the important subject of calcium orthophosphates. This type of materials is of special significance for human beings, because they represent the inorganic part of major normal (bones, teeth and antlers) and pathological (i.e., those appearing due to various diseases) calcified tissues of mammals. For example, atherosclerosis results in blood vessel blockage caused by a solid composite of cholesterol with calcium orthophosphates, while dental caries and osteoporosis mean a partial decalcification of teeth and bones, respectively, that results in replacement of a less soluble and harder biological apatite by more soluble and softer calcium hydrogenphosphates. Therefore, the processes of both normal and pathological calcifications are just an in vivo crystallization of calcium orthophosphates. Similarly, dental caries and osteoporosis might be considered an in vivo dissolution of calcium orthophosphates. Thus, calcium orthophosphates hold a great significance for humankind, and in this paper, an overview on the current knowledge on this subject is provided. PMID:23507744

  20. Calcium Hydroxylapatite

    PubMed Central

    Yutskovskaya, Yana Alexandrovna; Philip Werschler, WM.

    2015-01-01

    Background: Calcium hydroxylapatite is one of the most well-studied dermal fillers worldwide and has been extensively used for the correction of moderate-to-severe facial lines and folds and to replenish lost volume. Objectives: To mark the milestone of 10 years of use in the aesthetic field, this review will consider the evolution of calcium hydroxylapatite in aesthetic medicine, provide a detailed injection protocol for a global facial approach, and examine how the unique properties of calcium hydroxylapatite provide it with an important place in today’s market. Methods: This article is an up-to-date review of calcium hydroxylapatite in aesthetic medicine along with procedures for its use, including a detailed injection protocol for a global facial approach by three expert injectors. Conclusion: Calcium hydroxylapatite is a very effective agent for many areas of facial soft tissue augmentation and is associated with a high and well-established safety profile. Calcium hydroxylapatite combines high elasticity and viscosity with an ability to induce long-term collagen formation making it an ideal agent for a global facial approach. PMID:25610523

  1. Excessive Osteocytic Fgf23 Secretion Contributes to Pyrophosphate Accumulation and Mineralization Defect in Hyp Mice

    PubMed Central

    Murali, Sathish K.; Andrukhova, Olena; Clinkenbeard, Erica L.; White, Kenneth E.; Erben, Reinhold G.

    2016-01-01

    X-linked hypophosphatemia (XLH) is the most frequent form of inherited rickets in humans caused by mutations in the phosphate-regulating gene with homologies to endopeptidases on the X-chromosome (PHEX). Hyp mice, a murine homologue of XLH, are characterized by hypophosphatemia, inappropriately low serum vitamin D levels, increased serum fibroblast growth factor-23 (Fgf23), and osteomalacia. Although Fgf23 is known to be responsible for hypophosphatemia and reduced vitamin D hormone levels in Hyp mice, its putative role as an auto-/paracrine osteomalacia-causing factor has not been explored. We recently reported that Fgf23 is a suppressor of tissue nonspecific alkaline phosphatase (Tnap) transcription via FGF receptor-3 (FGFR3) signaling, leading to inhibition of mineralization through accumulation of the TNAP substrate pyrophosphate. Here, we report that the pyrophosphate concentration is increased in Hyp bones, and that Tnap expression is decreased in Hyp-derived osteocyte-like cells but not in Hyp-derived osteoblasts ex vivo and in vitro. In situ mRNA expression profiling in bone cryosections revealed a ~70-fold up-regulation of Fgfr3 mRNA in osteocytes versus osteoblasts of Hyp mice. In addition, we show that blocking of increased Fgf23-FGFR3 signaling with anti-Fgf23 antibodies or an FGFR3 inhibitor partially restored the suppression of Tnap expression, phosphate production, and mineralization, and decreased pyrophosphate concentration in Hyp-derived osteocyte-like cells in vitro. In vivo, bone-specific deletion of Fgf23 in Hyp mice rescued the suppressed TNAP activity in osteocytes of Hyp mice. Moreover, treatment of wild-type osteoblasts or mice with recombinant FGF23 suppressed Tnap mRNA expression and increased pyrophosphate concentrations in the culture medium and in bone, respectively. In conclusion, we found that the cell autonomous increase in Fgf23 secretion in Hyp osteocytes drives the accumulation of pyrophosphate through auto-/paracrine suppression

  2. Stone size limits the use of Hounsfield units for prediction of calcium oxalate stone composition.

    PubMed

    Stewart, Gregory; Johnson, Lewis; Ganesh, Halemane; Davenport, Daniel; Smelser, Woodson; Crispen, Paul; Venkatesh, Ramakrishna

    2015-02-01

    To evaluate the role of stone size in predicting urinary calculus composition using Hounsfield units on noncontrasted computed tomography (CT) scan. A retrospective review was performed for all patients who underwent ureteroscopy or percutaneous nephrolithotomy during a 1-year period, had a stone analysis performed, and had CT imaging available for review. All CT scans were reviewed by a board-certified radiologist. Variables evaluated included age, sex, body mass index, stone size, stone location, Hounsfield units (HUs), and stone composition. We identified a total of 91 patients (41 men and 50 women) with CT imaging and stone analysis available for review. Stone analysis showed 41 calcium oxalate monohydrate (CaOxMH), 13 calcium oxalate dihydrate, 29 calcium phosphate, 5 uric acid, 2 struvite, and 1 cystine stone. Average age was 46 years, and average body mass index was 32 kg/m2. Measured HUs varied significantly with size for CaOxMH and calcium oxalate dihydrate stones (P values <.05), but not for calcium phosphate stones (P = .126). Using a CaOxMH identification value of 700-1000 HUs, 28 of 41 stone compositions (68%) would not have been correctly identified, including all 10 (100%) small (<5 mm) stones, 13 of 22 (59%) medium (5-10 mm) stones, and 5 of 9 large (>10 mm) stones (55%). For calcium stones, the ability of CT HUs to predict stone composition was limited, likely due to the mixed stone composition. Within a cohort of CaOxMH stone formers, measured HUs varied linearly with stone size. All stones <5 mm were below thresholds for CaOxMH composition. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Pyrophosphate-mediated iron acquisition from transferrin in Neisseria meningitidis does not require TonB activity.

    PubMed

    Biville, Francis; Brézillon, Christophe; Giorgini, Dario; Taha, Muhamed-Kheir

    2014-01-01

    The ability to acquire iron from various sources has been demonstrated to be a major determinant in the pathogenesis of Neisseria meningitidis. Outside the cells, iron is bound to transferrin in serum, or to lactoferrin in mucosal secretions. Meningococci can extract iron from iron-loaded human transferrin by the TbpA/TbpB outer membrane complex. Moreover, N. meningitidis expresses the LbpA/LbpB outer membrane complex, which can extract iron from iron-loaded human lactoferrin. Iron transport through the outer membrane requires energy provided by the ExbB-ExbD-TonB complex. After transportation through the outer membrane, iron is bound by periplasmic protein FbpA and is addressed to the FbpBC inner membrane transporter. Iron-complexing compounds like citrate and pyrophosphate have been shown to support meningococcal growth ex vivo. The use of iron pyrophosphate as an iron source by N. meningitidis was previously described, but has not been investigated. Pyrophosphate was shown to participate in iron transfer from transferrin to ferritin. In this report, we investigated the use of ferric pyrophosphate as an iron source by N. meningitidis both ex vivo and in a mouse model. We showed that pyrophosphate was able to sustain N. meningitidis growth when desferal was used as an iron chelator. Addition of a pyrophosphate analogue to bacterial suspension at millimolar concentrations supported N. meningitidis survival in the mouse model. Finally, we show that pyrophosphate enabled TonB-independent ex vivo use of iron-loaded human or bovine transferrin as an iron source by N. meningitidis. Our data suggest that, in addition to acquiring iron through sophisticated systems, N. meningitidis is able to use simple strategies to acquire iron from a wide range of sources so as to sustain bacterial survival.

  4. Pyrophosphate-Mediated Iron Acquisition from Transferrin in Neisseria meningitidis Does Not Require TonB Activity

    PubMed Central

    Biville, Francis; Brézillon, Christophe; Giorgini, Dario; Taha, Muhamed-Kheir

    2014-01-01

    The ability to acquire iron from various sources has been demonstrated to be a major determinant in the pathogenesis of Neisseria meningitidis. Outside the cells, iron is bound to transferrin in serum, or to lactoferrin in mucosal secretions. Meningococci can extract iron from iron-loaded human transferrin by the TbpA/TbpB outer membrane complex. Moreover, N. meningitidis expresses the LbpA/LbpB outer membrane complex, which can extract iron from iron-loaded human lactoferrin. Iron transport through the outer membrane requires energy provided by the ExbB-ExbD-TonB complex. After transportation through the outer membrane, iron is bound by periplasmic protein FbpA and is addressed to the FbpBC inner membrane transporter. Iron-complexing compounds like citrate and pyrophosphate have been shown to support meningococcal growth ex vivo. The use of iron pyrophosphate as an iron source by N. meningitidis was previously described, but has not been investigated. Pyrophosphate was shown to participate in iron transfer from transferrin to ferritin. In this report, we investigated the use of ferric pyrophosphate as an iron source by N. meningitidis both ex vivo and in a mouse model. We showed that pyrophosphate was able to sustain N. meningitidis growth when desferal was used as an iron chelator. Addition of a pyrophosphate analogue to bacterial suspension at millimolar concentrations supported N. meningitidis survival in the mouse model. Finally, we show that pyrophosphate enabled TonB-independent ex vivo use of iron-loaded human or bovine transferrin as an iron source by N. meningitidis. Our data suggest that, in addition to acquiring iron through sophisticated systems, N. meningitidis is able to use simple strategies to acquire iron from a wide range of sources so as to sustain bacterial survival. PMID:25290693

  5. Biosynthesis of monoterpenes. Stereochemistry of the enzymatic cyclizations of geranyl pyrophosphate to (+)-alpha-pinene and (-)-beta-pinene

    SciTech Connect

    Croteau, R.; Satterwhite, D.M.; Wheeler, C.J.; Felton, N.M.

    1989-02-05

    The conversion of geranyl pyrophosphate to (+)-alpha-pinene and to (-)-beta-pinene is considered to proceed by the initial isomerization of the substrate to (-)-(3R)- and to (+)-(3S)-linalyl pyrophosphate, respectively, and the subsequent cyclization of the anti, endo-conformer of these bound intermediates by mirror-image sequences which should result in the net retention of configuration at C1 of the geranyl precursor. Incubation of (1R)-(2-14C,1-3H)- and (1S)-(2-14C,1-3H)geranyl pyrophosphate with (+)-pinene cyclase and with (-)-pinene cyclase from common sage (Salvia officinalis) gave labeled (+)-alpha- and (-)-beta-pinene of unchanged 3H/14C ratio in all cases, and the (+)- and (-)-olefins were stereoselectively converted to (+)- and (-)-borneol, respectively, which were oxidized to the corresponding (+)- and (-)-isomers of camphor, again without change in isotope ratio. The location of the tritium was determined in each case by stereoselective, base-catalyzed exchange of the exo-alpha-hydrogens of these derived ketones. The results indicated that the configuration at C1 of the substrate was retained in the enzymatic transformations to the (+)- and (-)-pinenes, which is entirely consistent with the syn-isomerization of geranyl pyrophosphate to linalyl pyrophosphate, transoid to cisoid rotation, and anti, endo-cyclization of the latter. The absolute stereochemical elements of the antipodal reaction sequences were confirmed by the selective enzymatic conversions of (3R)- and (3S)-1Z-(1-3H)linalyl pyrophosphate to (+)-alpha-pinene and (-)-beta-pinene, respectively, and by the location of the tritium in the derived camphors as before. The summation of the results fully defines the overall stereochemistry of the coupled isomerization and cyclization of geranyl pyrophosphate to the antipodal pinenes.

  6. Effects of phosphates on shellfish and on calcium carbonate crystallization in vitro. Final report

    SciTech Connect

    Wilbur, K.M.

    1986-07-17

    It has been known that inorganic phosphate inhibits the precipitation of calcium carbonate in artificial sea water. This work addresses the question of whether phosphate also affects the deposition of CaCO/sub 3/ in the exoskeletons of invertebrates. Tetrasodiumpyrophosphate and pentasodiumtripolyphosphate in concentrations of 15 ppM caused abnormality, mortality, and inhibition of shell deposition in trochophore larvae of the oyster Crassostrea. Inhibition of shell growth resulting from pollution at 15 ppM could be expected in Rangia with orthophosphate, tetrasodium pyrophosphate, and sodiumtripolyphosphate, in Helisoma with tetrasodium pyrophosphate, and pentasodium tripolyphosphate, and in larvae of Crassostea the relative inhibitory action of shell growth was tetrasodiumpyrophosphate > sodiumtripolyphosphate > sodium orthophosphate greater than or equal to sodium hexametaphosphate. 4 refs.

  7. Calcium and Calcium Supplements: Achieving the Right Balance

    MedlinePlus

    ... calcium. Common calcium supplements may be labeled as: Calcium carbonate (40 percent elemental calcium) Calcium citrate (21 percent ... forms of calcium supplements are carbonate and citrate. Calcium carbonate is cheapest and therefore often a good first ...

  8. Anhydrous ammonioguanidinium(2+) and dihydrated bis[aminoguanidinium(1+)] hexafluorosilicates: new co-products of preparing ferroelectric ammonioguanidinium(2+) hexafluorozirconate.

    PubMed

    Ross, C R; Bauer, M R; Nielson, R M; Abrahams, S C

    1999-04-01

    Ammonioguanidinium hexafluorosilicate, CH8N4(2+).SiF6(2-), and bis(aminoguanidinium) hexafluorosilicate dihydrate, 2CH7N4+.SiF6(2-).2H2O, are new materials formed as by-products in course of preparing ferroelectric CH8N4ZrF6 in the presence of glassware. Their structures were determined for comparison with the corresponding hexafluorozirconates. All atoms including the eight H atoms in the CH8N4(2+) cation and the seven H atoms in the CH7N4+ cation have been located and refined with wR(F2) = 0.0653, R = 0.0255, S = 1.146 and wR(F2) = 0.0745, R = 0.0301, S = 1.065, respectively. The N2C-N-N backbone of the 2+ cation is close to planarity, while that of the 1+ cation does not differ significantly from planarity. The SiF6(2-) octahedron is nearly ideally regular in both materials, with < Si-F > = 1.684 (unbiassed estimator of standard uncertainty = 0.016) A in the anhydrous hexafluorosilicate and 1.6801 (unbiassed estimator of standard uncertainty = 0.0006) A in the dihydrate. The combination of coulombic and NH...F interactions in CH8N4SiF6 results in a relatively dense variant of the NaCl structure. In addition to similar forces, the dihydrate is also characterized by the role of the water molecule with its strong NH...O interactions; its packing efficiency is, however, appreciably less than that of the anhydrous hexafluorosilicate with an approximately 8% increase in void space. Cleaved crystals of the dihydrate are frequently twinned across the (001) composition plane, with a twofold rotation about the b axis as the twin operation.

  9. Studies on the mechanism of pyrophosphate-mediated uptake of iron from transferrin by isolated rat-liver mitochondria.

    PubMed

    Konopka, K; Romslo, I

    1981-07-01

    1. Respiring rat liver mitochondria accumulate iron released from transferrin by pyrophosphate. The amount of iron accumulated is 1--1.5 nmol mg protein-1 h-1, or approximately 60% of the amount of iron mobilized from transferrin. 2. The uptake declines if respiration is inhibited, substrate is deleted, or the experiments are run under anaerobic conditions. Substrate depletion and respiratory inhibitors are less inhibitory under anaerobic conditions. 3. More than 80% of the amount of iron accumulated by aerobic, actively respiring mitochondria can be chelated by bathophenanthroline sulphonate, and with deuteroporphyrin included, up to 30% of the amount of iron accumulated is recovered as deuteroheme. Iron accumulated by respiration-inhibited mitochondria under aerobic conditions is not available for heme synthesis. 4. With time the uptake of iron increases eightfold relative to the uptake of pyrophosphate. 5. The results are compatible with a model in which ferric iron is mobilized from transferrin by pyrophosphate, ferric iron pyrophosphate is bound to the mitochondria, iron is reduced, dissociates from pyrophosphate and is taken up by the mitochondria. Ferrous iron thus formed is available for heme synthesis.

  10. Detection of Calcium Crystals in Knee Osteoarthritis Synovial Fluid: A Comparison Between Polarized Light and Scanning Electron Microscopy.

    PubMed

    Frallonardo, Paola; Oliviero, Francesca; Peruzzo, Luca; Tauro, Leonardo; Scanu, Anna; Galozzi, Paola; Ramonda, Roberta; Punzi, Leonardo

    2016-10-01

    The identification of calcium crystals in synovial fluid (SF) of patients with osteoarthritis (OA) represents an important step in understanding the role of these crystals in synovial inflammation and disease progression. This study aimed to investigate the presence of calcium pyrophosphate (CPP) and basic calcium phosphate (BCP) crystals in SF collected from patients with symptomatic knee OA by scanning electron microscopy (SEM) coupled to x-ray energy dispersive spectroscopy, compensated polarized light microscopy (CPLM), and alizarin red staining. Seventy-four patients with knee OA were included in the study. Synovial fluid samples were collected after arthrocentesis and examined under CPLM for the assessment of CPP crystals. Basic calcium phosphate crystals were evaluated by alizarin red staining. All the samples were examined by SEM. The concordance between the 2 techniques was evaluated by Cohen κ agreement coefficient. Calcium pyrophosphate and BCP crystals were found, respectively, in 23 (31.1%) and 13 (17.5%) of 74 OA SFs by SEM analysis. Calcium pyrophosphate crystals were identified in 23 (31.1%) of 74 samples by CPLM, whereas BCP crystals were suspected in 27 (36.4%) of 74 samples. According to κ coefficient, the concordance between CPLM and SEM was 0.83 for CPP, and that between alizarin red and SEM was 0.68 for BCP. The results of our study showed a high level of concordance between the 2 microscope techniques as regards CPP crystal identification and a lower agreement for BCP crystals. Although this finding highlights the difficulty in identifying BCP crystals by alizarin red staining, the use of SEM remains unsuitable to apply in the clinical setting. Because of the in vitro inflammatory effect of BCP crystals, further work on their analysis in SF could provide important information about the OA process.

  11. Studies on magnetic properties of chemically synthesized crystalline calcium ferrite nanoparticles

    SciTech Connect

    Debnath, A.; Bera, A.; Saha, B.; Chattopadhyay, K. K.

    2016-05-23

    Spinel-type ferrites have taken a very important role for modern electronic industry. Most of these ferrites exhibit low-loss dielectric properties, high resistivity, low eddy current and also high temperature ferromagnetism. Calcium ferrite is one such important metal oxide which is environmentally safe, chemically stable, low cost and greatly abundant. This outstanding material of calcium ferrite is synthesized by a simple chemical precipitation method using NaOH as the precipitating agent. Ferric chloride anhydrous (FeCl{sub 3}) and Calcium chloride dihydrate (CaCl{sub 2}.2H{sub 2}O) were used as iron and calcium sources respectively. The samples were heated at 200°C for 8h to obtain homogeneous powder of Calcium ferrite. The powders were characterized by using X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), Transmission electrical microscopy (TEM), and Fourier transform infrared spectroscopic (FTIR) measurements. The polycrystalline nature of the sample was confirmed by X-ray diffraction study. The magnetic properties of the sample were investigated by vibrating sample magnetometer (VSM) measurements. Magnetization curve of the prepared sample depicts that as synthesized calcium ferrite nanoparticles have saturation magnetic moment of 1.74 emu/g and the coercivity of 35.08 Oe with superparamagnetic behavior. The synthesized calcium ferrite nanoparticles with such magnetic properties will be a candidate material for different applications in electronics and exploring its functionality in the field of recently developing semiconductor device physics and spintronics.

  12. Studies on magnetic properties of chemically synthesized crystalline calcium ferrite nanoparticles

    NASA Astrophysics Data System (ADS)

    Debnath, A.; Bera, A.; Chattopadhyay, K. K.; Saha, B.

    2016-05-01

    Spinel-type ferrites have taken a very important role for modern electronic industry. Most of these ferrites exhibit low-loss dielectric properties, high resistivity, low eddy current and also high temperature ferromagnetism. Calcium ferrite is one such important metal oxide which is environmentally safe, chemically stable, low cost and greatly abundant. This outstanding material of calcium ferrite is synthesized by a simple chemical precipitation method using NaOH as the precipitating agent. Ferric chloride anhydrous (FeCl3) and Calcium chloride dihydrate (CaCl2.2H2O) were used as iron and calcium sources respectively. The samples were heated at 200°C for 8h to obtain homogeneous powder of Calcium ferrite. The powders were characterized by using X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), Transmission electrical microscopy (TEM), and Fourier transform infrared spectroscopic (FTIR) measurements. The polycrystalline nature of the sample was confirmed by X-ray diffraction study. The magnetic properties of the sample were investigated by vibrating sample magnetometer (VSM) measurements. Magnetization curve of the prepared sample depicts that as synthesized calcium ferrite nanoparticles have saturation magnetic moment of 1.74 emu/g and the coercivity of 35.08 Oe with superparamagnetic behavior. The synthesized calcium ferrite nanoparticles with such magnetic properties will be a candidate material for different applications in electronics and exploring its functionality in the field of recently developing semiconductor device physics and spintronics.

  13. Phase transformation of calcium phenyl phosphate in calcium hydroxyapatite

    SciTech Connect

    Tanaka, Hidekazu . E-mail: hidekazu@riko.shimane-u.ac.jp; Ibaraki, Koshiro; Uemura, Masao; Hino, Ryozi; Kandori, Kazuhiko; Ishikawa, Tatsuo

    2007-07-03

    Calcium phenyl phosphate (CaPP) was synthesized from a mixture of Ca(OH){sub 2} and phenyl phosphate (C{sub 6}H{sub 5}PO{sub 4}H{sub 2}) in an aqueous media. XRD pattern of CaPP exhibited five diffraction peaks at 2{theta} = 6.6, 13.3, 20.0, 26.8 and 33.7{sup o}. The d-spacing ratio of these peaks was ca. 1:1/2:1/3:1/4:1/5. The molar ratios of Ca/P and phenyl/P of CaPP were 1.0 and 0.92, respectively, and the chemical formula of the material was expressed as (C{sub 6}H{sub 5}PO{sub 4}){sub 0.92}(HPO{sub 4}){sub 0.08}Ca.1.3H{sub 2}O, similar to that of dicalcium phosphate dihydrate (CaHPO{sub 4}.2H{sub 2}O: DCPD). These results allowed us to infer that CaPP is composed of a multilayer alternating bilayer of phenyl groups of the phosphates and DCPD-like phase. The structure of the material was essentially not altered after aging at pH 9.0-11.0 and 85 deg. C in an aqueous media. While, after aging at pH {<=}8.0, the diffraction peaks of CaPP were suddenly weakened and disappeared at pH 7.0. Besides, new peaks due to calcium hydroxyapatite (Ca{sub 10}(PO{sub 4}){sub 6}(OH){sub 2}: Hap) appeared and their intensity was strengthened with decreasing the solution pH. TEM observation revealed that the Hap particles formed at pH 6.0 are fibrous with ca. 1.5 {mu}m in length and ca. 0.2 {mu}m in width. From these results, it is presumed that the layered CaPP was dissolved, hydrolyzed and reprecipitated to fibrous Hap particles at pH {<=}8.0 and 85 deg. C in aqueous media. This phase transformation of CaPP in Hap resembled to the formation mechanism of Hap in animal organism.

  14. Calcium cyanide

    Integrated Risk Information System (IRIS)

    Jump to main content . Integrated Risk Information System Recent Additions | Contact Us Search : All EPA IRIS • You are here : EPA Home • Research • Environmental Assessment • IRIS • IRIS Summaries Redirect Page As of September 28 , 2010 , the assessment summary for calcium cyanide is included in th

  15. Structural basis for gene regulation by a thiamine pyrophosphate-sensing riboswitch.

    PubMed

    Serganov, Alexander; Polonskaia, Anna; Phan, Anh Tuân; Breaker, Ronald R; Patel, Dinshaw J

    2006-06-29

    Riboswitches are metabolite-sensing RNAs, typically located in the non-coding portions of messenger RNAs, that control the synthesis of metabolite-related proteins. Here we describe a 2.05 angstroms crystal structure of a riboswitch domain from the Escherichia coli thiM mRNA that responds to the coenzyme thiamine pyrophosphate (TPP). TPP is an active form of vitamin B1, an essential participant in many protein-catalysed reactions. Organisms from all three domains of life, including bacteria, plants and fungi, use TPP-sensing riboswitches to control genes responsible for importing or synthesizing thiamine and its phosphorylated derivatives, making this riboswitch class the most widely distributed member of the metabolite-sensing RNA regulatory system. The structure reveals a complex folded RNA in which one subdomain forms an intercalation pocket for the 4-amino-5-hydroxymethyl-2-methylpyrimidine moiety of TPP, whereas another subdomain forms a wider pocket that uses bivalent metal ions and water molecules to make bridging contacts to the pyrophosphate moiety of the ligand. The two pockets are positioned to function as a molecular measuring device that recognizes TPP in an extended conformation. The central thiazole moiety is not recognized by the RNA, which explains why the antimicrobial compound pyrithiamine pyrophosphate targets this riboswitch and downregulates the expression of thiamine metabolic genes. Both the natural ligand and its drug-like analogue stabilize secondary and tertiary structure elements that are harnessed by the riboswitch to modulate the synthesis of the proteins coded by the mRNA. In addition, this structure provides insight into how folded RNAs can form precision binding pockets that rival those formed by protein genetic factors.

  16. Structural Basis for Gene Regulation by a Thiamine Pyrophosphate-Sensing Riboswitch

    SciTech Connect

    Serganov,A.; Polonskaia, A.; Phan, A.; Breaker, R.; Patel, D.

    2006-01-01

    Riboswitches are metabolite-sensing RNAs, typically located in the non-coding portions of messenger RNAs, that control the synthesis of metabolite-related proteins. Here we describe a 2.05 Angstroms crystal structure of a riboswitch domain from the Escherichia coli thiM mRNA4 that responds to the coenzyme thiamine pyrophosphate (TPP). TPP is an active form of vitamin B1, an essential participant in many protein-catalysed reactions. Organisms from all three domains of life including bacteria, plants and fungi, use TPP-sensing riboswitches to control genes responsible for importing or synthesizing thiamine and its phosphorylated derivatives, making this riboswitch class the most widely distributed member of the metabolite-sensing RNA regulatory system. The structure reveals a complex folded RNA in which one subdomain forms an intercalation pocket for the 4-amino-5-hydroxymethyl-2-methylpyrimidine moiety of TPP, whereas another subdomain forms a wider pocket that uses bivalent metal ions and water molecules to make bridging contacts to the pyrophosphate moiety of the ligand. The two pockets are positioned to function as a molecular measuring device that recognizes TPP in an extended conformation. The central thiazole moiety is not recognized by the RNA, which explains why the antimicrobial compound pyrithiamine pyrophosphate targets this riboswitch and downregulates the expression of thiamine metabolic genes. Both the natural ligand and its drug-like analogue stabilize secondary and tertiary structure elements that are harnessed by the riboswitch to modulate the synthesis of the proteins coded by the mRNA. In addition, this structure provides insight into how folded RNAs can form precision binding pockets that rival those formed by protein genetic factors.

  17. Pyrophosphate-Driven Proton Transport by Microsomal Membranes of Corn Coleoptiles 1

    PubMed Central

    Chanson, Alain; Fichmann, Jenny; Spear, David; Taiz, Lincoln

    1985-01-01

    Corn (Zea mays L. cv Trojan T929) coleoptile membranes were fractionated on isopycnic sucrose density gradients. Two peaks of ATP-driven H+-transport activity, corresponding to the previously characterized tonoplast (1.07 grams per cubic centimeter) and Golgi (1.13 grams per cubic centimeter) fractions (Chanson and Taiz, Plant Physiol 1985 78: 232-240) were localized. Coincident with these were two peaks of inorganic pyrophosphate (PPi)-driven H+-transport. At saturating (3 millimolar) concentrations of Mg2+:ATP, the rate of proton transport was further enhanced by the addition of 3 millimolar PPi, and the stimulation was additive, i.e. equal to the sum of the two added separately. The specific PPi analog, imidodiphosphate, antagonized PPi-driven H+-transport, but had no effect on ATP-driven transport. Moreover, PPi-dependent proton transport in both tonoplast-enriched and Golgi-enriched fractions was strongly promoted by 50 millimolar KNO3, unlike the ATP-dependent H+-pumps of the same membranes. Taken together, the results indicate that PPi-driven proton transport is mediated by specific membrane-bound H+-translocating pyrophosphatases. Both potassium and a permanent anion (NO3− > Cl−), were required for maximum activity. The PPi-driven proton pumps were totally inhibited by N,N′-dicyclohexylcarbodiimide, but were insensitive to 100 millimolar vanadate. The PPi concentration in coleoptile extracts was determined using an NADH oxidation assay system coupled to purified pyrophosphate:fructose 6-phosphate 1-phosphotransferase (EC 2.7.1.90). The total pyrophosphate content of corn coleoptiles was 20 nanomoles/gram fresh weight. Assuming a cytoplasmic location, the calculated PPi concentration is sufficient to drive proton transport at 20% of the maximum rate measured in vitro for the tonoplast-enriched fraction, and 10% of the maximum rate for the Golgi-enriched fraction. PMID:16664362

  18. Effect of thiamine pyrophosphate on ischemia-reperfusion induced oxidative damage in rat kidney

    PubMed Central

    Altuner, Durdu; Cetin, Nihal; Suleyman, Bahadir; Aslan, Zeynep; Hacimuftuoglu, Ahmet; Gulaboglu, Mine; Isaoglu, Neslihan; Demiryilmaz, Ismail; Suleyman, Halis

    2013-01-01

    Objectives: The biochemical effects of thiamine pyrophosphate on ischemia-reperfusion (IR) induced oxidative damage and DNA mutation in rat kidney tissue were investigated, and compared to thiamine. Materials and Methods: Rats were divided into four groups: Renal ischemia-reperfusion (RIR); thiamine pyrophosphate + RIR (TPRIR); thiamine + RIR (TRIR); and sham group (SG). Results: The results of biochemical experiments have shown that malondialdehyde (MDA) levels in rat kidney tissue after TRIR and TPRIR treatment were 7.2 ± 0.5 (P > 0.05) and 3.3 ± 0.3 (P < 0.0001) μmol/g protein, respectively. The MDA levels in the SG rat kidney tissue and in RIR group were 3.6 ± 0.2 (P < 0.0001) and 7.6 ± 0.6 μmol/g protein, respectively. Total glutathione (tGSH) levels in TRIR, TPRIR, SG, and RIR animal groups were 2.2 ± 0.3 (P > 0.05), 5.8 ± 0.4 (P < 0.0001), 6.2 ± 0.2 (P < 0.0001), and 1.7 ± 0.2 nmol/g protein, respectively. In the TRIR, TPRIR, SG, and RIR animal groups; 8-hydroxyguanine (8-OHGua)/Gua levels, which indicate mutagenic DNA, were 1.75 ± 0.12 (P > 0.05), 0.93 ± 0.1 (P < 0.0001), 0.85 ± 0.08 (P < 0.0001), and 1.93 ± 0.24 pmol/L, respectively. Conclusions: It has been shown that thiamine pyrophosphate prevents increase in mutagenic DNA in IR induced oxidative damage, whereas thiamine does not have this effect. PMID:24014907

  19. Thallium-201 versus technetium-99m pyrophosphate myocardial imaging in detection and evaluation of patients with acute myocardial infarction

    SciTech Connect

    Pitt, B.; Thrall, J.H.

    1980-12-18

    Thallium-201 myocardial imaging is of value in the early detection and evaluation of patients with suspected acute infarction. Thallium imaging may have a special value in characterizing patients with cardiogenic shock and in detecting patients at risk for subsequent infarction or death or death or both, before hospital discharge. Approximately 95 percent of pateints with transmural or nontransmural myocardial infarction can be detected with technetium-99m pyrophosphate myocardial imaging if the imaging is performed 24 to 72 hours after the onset of symptoms. Pyrophosphate imaging may have an important role in the evaluation of patients during the early follow-up period after hospital discharge from an episode of acute infarction. The finding of a persistently positive pyrophosphate image suggests a poor prognosis and is associated with a relatively large incidence of subsequent myocardial infarction and death.

  20. Structural Basis for Flip-Flop Action of Thiamin Pyrophosphate-dependent Enzymes Revealed by Human Pyruvate Dehydrogenase

    NASA Technical Reports Server (NTRS)

    Ciszak, Ewa M.; Korotchkina, Lioubov G.; Dominiak, Paulina M.; Sidhu, Sukdeep; Patel, Mulchand S.

    2003-01-01

    The derivative of vitamin B1, thiamin pyrophosphate, is a cofactor of enzymes performing catalysis in pathways of energy production. In alpha (sub 2) beta (sub 2)-heterotetrameric human pyruvate dehydrogenase, this cofactor is used to cleave the C(sup alpha) -C(=O) bond of pyruvate followed by reductive acetyl transfer to lipoyl-dihydrolipoamide acetyltransferase. The dynamic nonequivalence of two, otherwise chemically equivalent, catalytic sites has not yet been understood. To understand the mechanism of action of this enzyme, we determined the crystal structure of the holo-form of human pyruvate dehydrogenase at 1.95-Angstrom resolution. We propose a model for the flip-flop action of this enzyme through a concerted approximately 2-Angstrom shuttle-like motion of its heterodimers. Similarity of thiamin pyrophosphate binding in human pyruvate dehydrogenase with functionally related enzymes suggests that this newly defined shuttle-like motion of domains is common to the family of thiamin pyrophosphate-dependent enzymes.

  1. Structural Basis for Flip-Flop Action of Thiamin Pyrophosphate-dependent Enzymes Revealed by Human Pyruvate Dehydrogenase

    NASA Technical Reports Server (NTRS)

    Ciszak, Ewa M.; Korotchkina, Lioubov G.; Dominiak, Paulina M.; Sidhu, Sukdeep; Patel, Mulchand S.

    2003-01-01

    The derivative of vitamin B1, thiamin pyrophosphate, is a cofactor of enzymes performing catalysis in pathways of energy production. In alpha (sub 2) beta (sub 2)-heterotetrameric human pyruvate dehydrogenase, this cofactor is used to cleave the C(sup alpha) -C(=O) bond of pyruvate followed by reductive acetyl transfer to lipoyl-dihydrolipoamide acetyltransferase. The dynamic nonequivalence of two, otherwise chemically equivalent, catalytic sites has not yet been understood. To understand the mechanism of action of this enzyme, we determined the crystal structure of the holo-form of human pyruvate dehydrogenase at 1.95-Angstrom resolution. We propose a model for the flip-flop action of this enzyme through a concerted approximately 2-Angstrom shuttle-like motion of its heterodimers. Similarity of thiamin pyrophosphate binding in human pyruvate dehydrogenase with functionally related enzymes suggests that this newly defined shuttle-like motion of domains is common to the family of thiamin pyrophosphate-dependent enzymes.

  2. Cyclization of farnesyl pyrophosphate to the sesquiterpene olefins humulene and caryophyllene by an enzyme system from sage (Salvia officinalis)

    SciTech Connect

    Croteau, R.; Gundy, A.

    1984-09-01

    A soluble enzyme preparation obtained from sage (Salvia officinalis) leaves was shown to catalyze the divalent metal-ion dependent cyclization of trans, trans-farnesyl pyrophosphate to the macrocyclic sesquiterpene olefins humulene and caryophyllene. The identities of the biosynthetic products were confirmed by radiochromatographic analysis and by preparation of crystalline derivatives, and the specificity of labeling in the cyclization reaction was established by chemical degradation of the olefins derived enzymatically from (1-3H2)farnesyl pyrophosphate. These results constitute the first report on the cyclization of farnesyl pyrophosphate to humulene and caryophyllene, two of the most common sesquiterpenes in nature, and the first description of a soluble sesquiterpene cyclase to be isolated from leaves of a higher plant.

  3. Rapid efflux of Ca2+ from heart mitochondria in the presence of inorganic pyrophosphate.

    PubMed

    Vercesi, A; Lehninger, A L

    1984-01-13

    Inorganic pyrophosphate (PPi) in the intracellular concentration range causes rapid efflux of Ca2+ from rat heart mitochondria oxidizing pyruvate + malate in a low Na+ medium. Half-maximal rates of Ca2+ efflux were given by 20 microM PPi. During and after PPi-stimulated Ca2+ efflux the mitochondria retain their structural integrity and complete respiratory control. Carboxyatractyloside inhibits PPi-stimulated Ca2+ efflux, indicating PPi must enter the matrix in order to promote Ca2+ efflux. Heart mitochondria have a much higher affinity for PPi uptake and PPi-induced Ca2+ efflux than liver mitochondria.

  4. Structure of the eukaryotic thiamine pyrophosphate riboswitch with its regulatory ligand.

    PubMed

    Thore, Stéphane; Leibundgut, Marc; Ban, Nenad

    2006-05-26

    Riboswitches are untranslated regions of messenger RNA, which adopt alternate structures depending on the binding of specific metabolites. Such conformational switching regulates the expression of proteins involved in the biosynthesis of riboswitch substrates. Here, we present the 2.9 angstrom-resolution crystal structure of the eukaryotic Arabidopsis thaliana thiamine pyrophosphate (TPP)-specific riboswitch in complex with its natural ligand. The riboswitch specifically recognizes the TPP via conserved residues located within two highly distorted parallel "sensor" helices. The structure provides the basis for understanding the reorganization of the riboswitch fold upon TPP binding and explains the mechanism of resistance to the antibiotic pyrithiamine.

  5. N-glycosylation in the thermoacidophilic archaeon Sulfolobus acidocaldarius involves a short dolichol pyrophosphate carrier.

    PubMed

    Guan, Ziqiang; Delago, Antonia; Nußbaum, Phillip; Meyer, Benjamin; Albers, Sonja-Verena; Eichler, Jerry

    2016-09-01

    N-glycosylation is a post-translational modification that occurs across evolution. In the thermoacidophilic archaea Sulfolobus acidocaldarius, glycoproteins are modified by an N-linked tribranched hexasaccharide reminiscent of the N-glycans assembled in Eukarya. Previously, hexose-bearing dolichol phosphate was detected in a S. acidocaldarius Bligh-Dyer lipid extract. Here, we used a specialized protocol for extracting lipid-linked oligosaccharides to detect a dolichol pyrophosphate bearing the intact hexasaccharide, as well as its biosynthetic intermediates. Furthermore, evidence for N-glycosylation of two S. acidocaldarius proteins by the same hexasaccharide and its derivatives was collected. These findings thus provide novel insight into archaeal N-glycosylation.

  6. Single- and repeated-dose oral toxicity studies of citicoline free-base (choline cytidine 5'-pyrophosphate) in Sprague-Dawley rats.

    PubMed

    Schauss, A G; Somfai-Relle, S; Financsek, I; Glavits, R; Parent, S C; Endres, J R; Varga, T; Szücs, Z; Clewell, A

    2009-01-01

    The dietary supplement Citicoline free-base (choline cytidine 5'-pyrophosphate) was toxicologically evaluated in Sprague-Dawley rats using oral gavage. In an acute 14-day study, 2000 mg/kg was well tolerated. In a 90-day study, 100, 350, and 1000 mg/kg/day doses resulted in no mortality. In males, slight significant increases in serum creatinine (350 and 1000 mg/kg/day), and decreases in urine volume (all treated groups) were observed. In females, slight significant increases in total white blood cell and absolute lymphocyte counts (1000 mg/kg/day), and blood urea nitrogen (BUN) (100 and 350, but not 1000 mg/kg/day) were noted. A dose-related increase in renal tubular mineralization, without degenerative or inflammatory reaction, was found in females (all treated groups) and two males (1000 mg/kg/day). Renal mineralization in rats (especially females) is influenced by calcium:phosphorus ratios in the diet. A high level of citicoline consumption resulted in increased phosphorus intake in the rats, and likely explains this result.

  7. Involvement of the pyrophosphate and the 2'-phosphate binding regions of ferredoxin-NADP+ reductase in coenzyme specificity.

    PubMed

    Tejero, Jesús; Martínez-Julvez, Marta; Mayoral, Tomas; Luquita, Alejandra; Sanz-Aparicio, Julia; Hermoso, Juan A; Hurley, John K; Tollin, Gordon; Gómez-Moreno, Carlos; Medina, Milagros

    2003-12-05

    Previous studies indicated that the determinants of coenzyme specificity in ferredoxin-NADP+ reductase (FNR) from Anabaena are situated in the 2'-phosphate (2'-P) NADP+ binding region, and also suggested that other regions must undergo structural rearrangements of the protein backbone during coenzyme binding. Among the residues involved in such specificity could be those located in regions where interaction with the pyrophosphate group of the coenzyme takes place, namely loops 155-160 and 261-268 in Anabaena FNR. In order to learn more about the coenzyme specificity determinants, and to better define the structural basis of coenzyme binding, mutations in the pyrophosphate and 2'-P binding regions of FNR have been introduced. Modification of the pyrophosphate binding region, involving residues Thr-155, Ala-160, and Leu-263, indicates that this region is involved in determining coenzyme specificity and that selected alterations of these positions produce FNR enzymes that are able to bind NAD+. Thus, our results suggest that slightly different structural rearrangements of the backbone chain in the pyrophosphate binding region might determine FNR specificity for the coenzyme. Combined mutations at the 2'-P binding region, involving residues Ser-223, Arg-224, Arg-233, and Tyr-235, in combination with the residues mentioned above in the pyrophosphate binding region have also been carried out in an attempt to increase the FNR affinity for NAD+/H. However, in most cases the analyzed mutants lost the ability for NADP+/H binding and electron transfer, and no major improvements were observed with regard to the efficiency of the reactions with NAD+/H. Therefore, our results confirm that determinants for coenzyme specificity in FNR are also situated in the pyrophosphate binding region and not only in the 2'-P binding region. Such observations also suggest that other regions of the protein, yet to be identified, might also be involved in this process.

  8. Reduction of orthophosphates loss in agricultural soil by nano calcium sulfate.

    PubMed

    Chen, Dong; Szostak, Paul; Wei, Zongsu; Xiao, Ruiyang

    2016-01-01

    Nutrient loss from soil, especially phosphorous (P) from farmlands to natural water bodies via surface runoff or infiltration, have caused significant eutrophication problems. This is because dissolved orthophosphates are usually the limiting nutrient for algal blooms. Currently, available techniques to control eutrophication are surprisingly scarce. Calcium sulfate or gypsum is a common soil amendment and has a strong complexation to orthophosphates. The results showed that calcium sulfate reduced the amount of water extractable P (WEP) through soil incubation tests, suggesting less P loss from farmlands. A greater decrease in WEP occurred with a greater dosage of calcium sulfate. Compared to conventional coarse calcium sulfate, nano calcium sulfate further reduced WEP by providing a much greater specific surface area, higher solubility, better contact with the fertilizer and the soil particles, and superior dispersibility. The enhancement of the nano calcium sulfate for WEP reduction is more apparent for a pellet- than a powdered- fertilizer. At the dosage of Ca/P weight ratio of 2.8, the WEP decreased by 31±5% with the nano calcium sulfate compared to 20±5% decrease with the coarse calcium sulfate when the pellet fertilizer was used. Computation of the chemical equilibrium speciation shows that calcium hydroxyapatite has the lowest solubility. However, other mineral phases such as hydroxydicalcium phosphate, dicalcium phosphate dihydrate, octacalcium phosphate, and tricalcium phosphate might form preceding to calcium hydroxyapatite. Since calcium sulfate is the major product of the flue gas desulfurization (FGD) process, this study demonstrates a potential beneficial reuse and reduction of the solid FGD waste. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. A colorimetric assay for inorganic pyrophosphate that is also useful for measuring product accumulation in polymerase chain reactions.

    PubMed

    Tagiri-Endo, Misako

    2003-04-15

    A novel coupled enzyme assay for measuring inorganic pyrophosphate (PP(i)) in biological samples is described. The total PP(i) is determined by a reaction with inosine 5'-monophosphate, catalyzed by hypoxanthine-guanine phosphoribosyl transferase, yielding hypoxanthine and phosphoribosyl pyrophosphate. The hypoxanthine is oxidized to uric acid by xanthine oxidase/xanthine dehydrogenase and can be measured by formation of formazan when a tetrazolium salt is used as the oxidant. The method is also useful for detecting and quantifying PP(i) released from nucleotides during polymerase chain reactions. This rapid and simple method for detecting amplified nucleic acids permits low-cost monitoring by eye or spectrophotometer.

  10. Synthesis of calcium oxalate crystals in culture medium irradiated with non-equilibrium atmospheric-pressure plasma

    NASA Astrophysics Data System (ADS)

    Kurake, Naoyuki; Tanaka, Hiromasa; Ishikawa, Kenji; Nakamura, Kae; Kajiyama, Hiroaki; Kikkawa, Fumitaka; Mizuno, Masaaki; Yamanishi, Yoko; Hori, Masaru

    2016-09-01

    Octahedral particulates several tens of microns in size were synthesized in a culture medium irradiated through contact with a plume of non-equilibrium atmospheric-pressure plasma (NEAPP). The particulates were identified in the crystalline phase as calcium oxalate dihydrate (COD). The original medium contained constituents such as NaCl, d-glucose, CaCl2, and NaHCO3 but not oxalate or oxalic acid. The oxalate was clearly synthesized and crystallized in the medium as thermodynamically unstable COD crystals after the NEAPP irradiation.

  11. On 2:1 melamine - Squaric acid dihydrate complex: The structure and vibrational spectra

    NASA Astrophysics Data System (ADS)

    Nowicka-Scheibe, J.; Pawlukojć, A.; Sobczyk, L.; Jański, J.

    2017-01-01

    In the present paper we would like to describe the structural and dynamical properties of crystalline dihydrated complex of melamine (2,4,6-triamino-1,3,5-triazin-1-ium) with squaric acid (3,4-dihydroxycyclobut-3-ene-1,2-dione) abbreviated as MH·SQ. The X-ray diffraction studies show the presence of deprotonated units (C4O4)2- and single protonated melamine cations surrounded by tetrameric water assemblies (H2O)4. The formation of the water tetramers deserves a special attention. IR absorption and Raman spectra reflect a richness of structural units and numerous hydrogen bonds. The presence of the continua in the IR spectra, with a characteristic presence of the Hadži's trio enriched by a numerous submaxima, may be ascribed to the structural units and to the various types of hydrogen bonds. The density functional theory calculation with the periodic boundary conditions was use to precise analysis of experimental data.

  12. Synthesis and bioactivities of silver nanoparticles capped with 5-Amino-?-resorcylic acid hydrochloride dihydrate

    PubMed Central

    2014-01-01

    Background Conjugated and drug loaded silver nanoparticles are getting an increased attention for various biomedical applications. Nanoconjugates showed significant enhancement in biological activity in comparison to free drug molecules. In this perspective, we report the synthesis of bioactive silver capped with 5-Amino-?-resorcylic acid hydrochloride dihydrate (AR). The in vitro antimicrobial (antibacterial, antifungal), enzyme inhibition (xanthine oxidase, urease, carbonic anhydrase, ?-chymotrypsin, cholinesterase) and antioxidant activities of the developed nanostructures was investigated before and after conjugation to silver metal. Results The conjugation of AR to silver was confirmed through FTIR, UV¿vis and TEM techniques. The amount of AR conjugated with silver was characterized through UV¿vis spectroscopy and found to be 9% by weight. The stability of synthesized nanoconjugates against temperature, high salt concentration and pH was found to be good. Nanoconjugates, showed significant synergic enzyme inhibition effect against xanthine and urease enzymes in comparison to standard drugs, pure ligand and silver. Conclusions Our synthesized nanoconjugate was found be to efficient selective xanthine and urease inhibitors in comparison to Ag and AR. On a per weight basis, our nanoconjugates required less amount of AR (about 11 times) for inhibition of these enzymes. PMID:25201390

  13. Molecular structure, vibrational spectra and DFT computational studies of melaminium N-acetylglycinate dihydrate

    NASA Astrophysics Data System (ADS)

    Tanak, H.; Pawlus, K.; Marchewka, M. K.

    2016-10-01

    Melaminium N-acetylglycinate dihydrate, an organic material has been synthesized and characterized by X-ray diffraction, FT-IR, and FT-Raman spectroscopies for the protiated and deuteriated crystals. The title complex crystallizes in the triclinic system, and the space group is P-1 with a = 5.642(1) Å, b = 7.773(2) Å, c = 15.775(3) Å, α = 77.28(1)°, β = 84.00(1)°, γ = 73.43(1)° and Z = 2. The molecular geometry, vibrational frequencies and intensity of the vibrational bands have been interpreted with the aid of structure optimization based on density functional method (B3LYP) with the 6-311++G(d,p) basis set. The obtained vibrational wavenumbers and optimized geometric parameters were seen to be in good agreement with the experimental data. The intermolecular hydrogen bonding interactions of the title compound have been investigated using the natural bonding orbital analysis. It reveals that the O-H···O, N-H···N and N-H···O intermolecular interactions significantly influence crystal packing of this molecule. The non-linear optical properties are also addressed theoretically. The predicted NLO properties of the title compound are much greater than ones of urea. In addition, DFT calculations of the title compound, molecular electrostatic potential, thermodynamic properties, frontier orbitals and chemical reactivity descriptors were also performed at 6-311++G(d,p) level of theory.

  14. Influence of an organic and an inorganic additive on the crystallization of dicalcium phosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Anee, T. K.; Meenakshi Sundaram, N.; Arivuoli, D.; Ramasamy, P.; Narayana Kalkura, S.

    2005-12-01

    Dicalcium phosphate dihydrate (DCPD) was crystallized by single diffusion method under physiological pH, in the presence of cobalt and malic acid. The morphology, composition and microstructure of the grown crystals were analyzed using EDTA titration, UV-Visible, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), atomic absorption spectroscopy (AAS) and scanning electron microscopy (SEM). These analyses showed that the grown crystals were Co 2+-doped DCPD. The temperature has influenced the dopant entry into DCPD crystals, but the dopant content and temperature were having not much influence on the crystal morphology. In pathological crystallization, the size and morphology are very important since they decide the mode of treatment to adopt. The morphological changes arise when the growth assay is doped with cobalt and malic acid. The effect of malic acid on the crystallization was highly specific, adsorbing on certain crystal faces during growth and producing different morphologies. At higher concentration, the morphology showed a feature frequently encountered in biomineralization, the orientational ordering in [0 0 1], leading to elongated crystals.

  15. Characterization of dicalcium phosphate dihydrate cements prepared using a novel hydroxyapatite-based formulation.

    PubMed

    Alge, Daniel L; Santa Cruz, Grace; Goebel, W Scott; Chu, Tien-Min Gabriel

    2009-04-01

    Dicalcium phosphate dihydrate (DCPD) cements are typically prepared using beta-tricalcium phosphate (beta-TCP) as the base component. However, hydroxyapatite (HA) is an interesting alternative because of its potential for reducing cement acidity, as well as modulating cement properties via ionic substitutions. In the present study, we have characterized DCPD cements prepared with a novel formulation based on monocalcium phosphate monohydrate (MCPM) and HA. Cements were prepared using a 4:1 MCPM:HA molar ratio. The reactivity of HA in this system was verified by showing DCPD formation using poorly crystalline HA, as well as highly crystalline HA. Evaluation of cements prepared with poorly crystalline HA revealed that setting occurs rapidly in the MCPM/HA system, and that the use of a setting regulator is necessary to maintain workability of the cement paste. Compressive testing showed that MCPM/HA cements have strengths comparable to what has previously been published for DCPD cements. However, preliminary in vitro analysis of cement degradation revealed that conversion of DCPD to HA may occur much more rapidly in the MCPM/HA system compared to cements prepared with beta-TCP. Future studies should investigate this property further, as it could have important implications for the use of HA-based DCPD cement formulations.

  16. Channel-like crystal structure of cinchoninium L-O-phosphoserine salt dihydrate.

    PubMed

    Wesełucha-Birczyńska, Aleksandra; Oleksyn, Barbara J; Watroba, Joanna

    2010-06-01

    Studies on the interactions between L-O- phosphoserine, as one of the simplest fragments of membrane components, and the Cinchona alkaloid cinchonine, in the crystalline state were performed. Cinchoninium L-O-phosposerine salt dihydrate (PhSerCin) crystallizes in a monoclinic crystal system, space group P2(1), with unit cell parameters: a = 8.45400(10) A, b = 7.17100(10) A, c = 20.7760(4) A, alpha = 90 degrees , beta = 98.7830(10) degrees , gamma = 90 degrees , Z = 2. The asymmetric unit consists of the cinchoninium cation linked by hydrogen bonds to a phosphoserine anion and two water molecules. Intermolecular hydrogen bonds connecting phosphoserine anions via water molecules form chains extended along the b axis. Two such chains symmetrically related by twofold screw axis create a "channel." On both sides of this channel cinchonine cations are attached by hydrogen bonds in which the atoms N1, O12, and water molecules participate. This arrangement mimics the system of bilayer biological membrane.

  17. Biodegradation behavior and cytotoxicity of the composite membrane composed of beta-dicalcium pyrophosphate and glucose mediated (polyethylene glycol/chitosan).

    PubMed

    Wang, Jian Wen; Hon, Min Hsiung

    2004-02-01

    The purpose of this study is to prepare and evaluate the biodegradation behavior and cytotoxicity of a composite membrane, G-beta-DCP, combining beta-dicalcium pyrophosphate (beta-DCP) ceramic particles and glucose mediated chitosan-polyethylene glycol (PEG) membrane. The cytotoxicity of the G-beta-DCP was examined by the in vitro method of NIH 3T3 fibroblast cell culture. Extracts were obtained by soaking the G-beta-DCP composite in lysozyme containing phosphate buffer solution for 2, 7, 14, 21 and 28 days, respectively. The substances released from the G-beta-DCP composite were analyzed by gas chromatography-mass spectrometry (GC-MAS) and inductively coupled plasma atomic emission spectrometry (ICP-AES). The change in morphologies, chemical composition and crystal structure was examined by scanning electron microscopy (SEM) and X-ray diffraction pattern (XRD). The results of extracts cocultured with fibroblasts show that the growth of fibroblasts would increase for the extracts obtained from different beta-DCP feeding weight G-beta-DCP composites after soaking for 7 days. After further increasing the soaking time, the cell number still increases. It is found that the glucose amine and calcium are gradually released from the G-beta-DCP composites, which is considered to be nutritious for the growth of the fibroblast. The release rate of calcium ion and glucosamine concentration can be regulated by feeding the beta-DCP. The degradation behavior of G-beta-DCP composite is considered as an "onion degradation model" that the G-beta-DCP degrades from outer layer to inner layer. The developed material should have a great potential as a cell substrate in the field of tissue engineering.

  18. Cloning and characterization of farnesyl pyrophosphate synthase from the highly branched isoprenoid producing diatom Rhizosolenia setigera

    PubMed Central

    Ferriols, Victor Marco Emmanuel N.; Yaginuma, Ryoko; Adachi, Masao; Takada, Kentaro; Matsunaga, Shigeki; Okada, Shigeru

    2015-01-01

    The diatom Rhizosolenia setigera Brightwell produces highly branched isoprenoid (HBI) hydrocarbons that are ubiquitously present in marine environments. The hydrocarbon composition of R. setigera varies between C25 and C30 HBIs depending on the life cycle stage with regard to auxosporulation. To better understand how these hydrocarbons are biosynthesized, we characterized the farnesyl pyrophosphate (FPP) synthase (FPPS) enzyme of R. setigera. An isolated 1465-bp cDNA clone contained an open reading frame spanning 1299-bp encoding a protein with 432 amino acid residues. Expression of the RsFPPS cDNA coding region in Escherichia coli produced a protein that exhibited FPPS activity in vitro. A reduction in HBI content from diatoms treated with an FPPS inhibitor, risedronate, suggested that RsFPPS supplies precursors for HBI biosynthesis. Product analysis by gas chromatography-mass spectrometry also revealed that RsFPPS produced small amounts of the cis-isomers of geranyl pyrophosphate and FPP, candidate precursors for the cis-isomers of HBIs previously characterized. Furthermore, RsFPPS gene expression at various life stages of R. setigera in relation to auxosporulation were also analyzed. Herein, we present data on the possible role of RsFPPS in HBI biosynthesis, and it is to our knowledge the first instance that an FPPS was cloned and characterized from a diatom. PMID:25996801

  19. Nuclear receptor engineering based on novel structure activity relationships revealed by farnesyl pyrophosphate

    PubMed Central

    Goyanka, Ritu; Das, Sharmistha; Samuels, Herbert H.; Cardozo, Timothy

    2010-01-01

    Nuclear receptors (NRs) comprise the second largest protein family targeted by currently available drugs, acting via specific ligand interactions within the ligand binding domain (LBD). Recently, farnesyl pyrophosphate (FPP) was shown to be a unique promiscuous NR ligand, activating a subset of NR family members and inhibiting wound healing in skin. The current study aimed at visualizing the unique basis of FPP interaction with multiple receptors in order to identify general structure–activity relationships that operate across the NR family. Docking of FPP to the 3D structures of the LBDs of a diverse set of NRs consistently revealed an electrostatic FPP pyrophosphate contact with an NR arginine conserved in the NR family, a hydrophobic farnesyl contact with NR helix-12 and a ligand binding pocket volume between 300 and 430 Å3 as the minimal requirements for FPP activation of any NR. Lack of any of these structural features appears to render a given NR resistant to FPP activation. We used these structure–activity relationships to rationally design and successfully engineer several mutant human estrogen receptors that retain responsiveness to estradiol but no longer respond to FPP. PMID:20817759

  20. Nuclear receptor engineering based on novel structure activity relationships revealed by farnesyl pyrophosphate.

    PubMed

    Goyanka, Ritu; Das, Sharmistha; Samuels, Herbert H; Cardozo, Timothy

    2010-11-01

    Nuclear receptors (NRs) comprise the second largest protein family targeted by currently available drugs, acting via specific ligand interactions within the ligand binding domain (LBD). Recently, farnesyl pyrophosphate (FPP) was shown to be a unique promiscuous NR ligand, activating a subset of NR family members and inhibiting wound healing in skin. The current study aimed at visualizing the unique basis of FPP interaction with multiple receptors in order to identify general structure-activity relationships that operate across the NR family. Docking of FPP to the 3D structures of the LBDs of a diverse set of NRs consistently revealed an electrostatic FPP pyrophosphate contact with an NR arginine conserved in the NR family, a hydrophobic farnesyl contact with NR helix-12 and a ligand binding pocket volume between 300 and 430 Å(3) as the minimal requirements for FPP activation of any NR. Lack of any of these structural features appears to render a given NR resistant to FPP activation. We used these structure-activity relationships to rationally design and successfully engineer several mutant human estrogen receptors that retain responsiveness to estradiol but no longer respond to FPP.

  1. Cloning and characterization of farnesyl pyrophosphate synthase from the highly branched isoprenoid producing diatom Rhizosolenia setigera.

    PubMed

    Ferriols, Victor Marco Emmanuel N; Yaginuma, Ryoko; Adachi, Masao; Takada, Kentaro; Matsunaga, Shigeki; Okada, Shigeru

    2015-05-21

    The diatom Rhizosolenia setigera Brightwell produces highly branched isoprenoid (HBI) hydrocarbons that are ubiquitously present in marine environments. The hydrocarbon composition of R. setigera varies between C25 and C30 HBIs depending on the life cycle stage with regard to auxosporulation. To better understand how these hydrocarbons are biosynthesized, we characterized the farnesyl pyrophosphate (FPP) synthase (FPPS) enzyme of R. setigera. An isolated 1465-bp cDNA clone contained an open reading frame spanning 1299-bp encoding a protein with 432 amino acid residues. Expression of the RsFPPS cDNA coding region in Escherichia coli produced a protein that exhibited FPPS activity in vitro. A reduction in HBI content from diatoms treated with an FPPS inhibitor, risedronate, suggested that RsFPPS supplies precursors for HBI biosynthesis. Product analysis by gas chromatography-mass spectrometry also revealed that RsFPPS produced small amounts of the cis-isomers of geranyl pyrophosphate and FPP, candidate precursors for the cis-isomers of HBIs previously characterized. Furthermore, RsFPPS gene expression at various life stages of R. setigera in relation to auxosporulation were also analyzed. Herein, we present data on the possible role of RsFPPS in HBI biosynthesis, and it is to our knowledge the first instance that an FPPS was cloned and characterized from a diatom.

  2. Molecular Characterization of a Novel Geranylgeranyl Pyrophosphate Synthase from Plasmodium Parasites*

    PubMed Central

    Artz, Jennifer D.; Wernimont, Amy K.; Dunford, James E.; Schapira, Matthieu; Dong, Aiping; Zhao, Yong; Lew, Jocelyne; Russell, R. Graham G.; Ebetino, F. Hal; Oppermann, Udo; Hui, Raymond

    2011-01-01

    We present here a study of a eukaryotic trans-prenylsynthase from the malaria pathogen Plasmodium vivax. Based on the results of biochemical assays and contrary to previous indications, this enzyme catalyzes the production of geranylgeranyl pyrophosphate (GGPP) rather than farnesyl pyrophosphate (FPP). Structural analysis shows that the product length is constrained by a hydrophobic cavity formed primarily by a set of residues from the same subunit as the product as well as at least one other from the dimeric partner. Furthermore, Plasmodium GGPP synthase (GGPPS) can bind nitrogen-containing bisphosphonates (N-BPs) strongly with the energetically favorable cooperation of three Mg2+, resulting in inhibition by this class of compounds at IC50 concentrations below 100 nm. In contrast, human and yeast GGPPSs do not accommodate a third magnesium atom in the same manner, resulting in their insusceptibility to N-BPs. This differentiation is in part attributable to a deviation in a conserved motif known as the second aspartate-rich motif: whereas the aspartates at the start and end of the five-residue motif in FFPP synthases and P. vivax GGPPSs both participate in the coordination of the third Mg2+, an asparagine is featured as the last residue in human and yeast GGPPSs, resulting in a different manner of interaction with nitrogen-containing ligands. PMID:21084289

  3. Molecular cloning, characterization, and function analysis of a mevalonate pyrophosphate decarboxylase gene from Ganoderma lucidum.

    PubMed

    Shi, Liang; Qin, Lei; Xu, Yingjie; Ren, Ang; Fang, Xing; Mu, Dashuai; Tan, Qi; Zhao, Mingwen

    2012-05-01

    This study investigated the role of the mevalonate pyrophosphate decarboxylase gene in the triterpene biosynthetic pathway of Ganoderma lucidum. The mevalonate pyrophosphate decarboxylase gene (mvd) was isolated using a degenerate primer-PCR technique. An analysis of the Gl-mvd transcription profile revealed a positive correlation between the expression of the Gl-mvd gene and triterpene content changes in G. lucidum during development. Furthermore, a promoter deletion analysis was conducted in G. lucidum to investigate the promoter activity and the role of methyl jasmonate (MeJA) responsive elements in the mvd promoter under the MeJA elicitor. The overexpression of Gl-mvd increased triterpene accumulation compared with the wild-type strain and increased the expression of several genes involved in the triterpene biosynthetic pathway. The findings of this study suggest that mvd may play an important role in triterpene biosynthesis regulation. Moreover, there may be the interactions among the genes involved in the triterpene biosynthetic pathway in the G. lucidum. Additionally, this study provides an approach for improving triterpene content through the overexpression of a key gene.

  4. Phosphate-group recognition by the aptamer domain of the thiamine pyrophosphate sensing riboswitch.

    PubMed

    Noeske, Jonas; Richter, Christian; Stirnal, Elke; Schwalbe, Harald; Wöhnert, Jens

    2006-09-01

    Riboswitches are highly structured RNA elements that control gene expression by binding directly to small metabolite molecules. Remarkably, many of these metabolites contain negatively charged phosphate groups that contribute significantly to the binding affinity. An example is the thiamine pyrophosphate-sensing riboswitch in the 5'-untranslated region of the E. coli thiM mRNA. This riboswitch binds, in order of decreasing affinity, to thiamine pyrophosphate (TPP), thiamine monophosphate (TMP), and thiamine, which contain two, one, and no phosphate groups, respectively. We examined the binding of TPP and TMP to this riboswitch by using (31)P NMR spectroscopy. Chemical-shift changes were observed for the alpha- and beta-phosphate group of TPP and the phosphate group of TMP upon RNA binding; this indicates that they are in close contact with the RNA. Titration experiments with paramagnetic Mn(2+) ions revealed strong line-broadening effects for both (31)P signals of the bound TPP; this indicates a Mg(2+) binding site in close proximity and suggests that the phosphate group(s) of the ligand is/are recognized in a magnesium ion-mediated manner by the RNA.

  5. Thiamine pyrophosphate riboswitch in some representative plant species: a bioinformatics study.

    PubMed

    Yadav, Sunita; Swati, D; Chandrasekharan, Hariharan

    2015-01-01

    Metabolites regulate their own production by directly interacting with highly conserved regions of mRNA that are capable of forming discrete tertiary structures. Such regions of mRNA are called riboswitches. The thiamine pyrophosphate (TPP) riboswitch is the most common riboswitch in different organisms. The TPP is an essential coenzyme that is synthesized by the coupling of pyrimidine (hydroxymethyl pyrimidine) and thiazole (hydroxyethyl thiazole). The TPP riboswitch was searched across all possible phyla of plant kingdom by using Arabidopsis thaliana, a model organism in which TPP riboswitch is already found. The aptameric domain of the TPP riboswitch is conserved at the sequence as well as structure levels in all chosen plant species. The sequence of the noncoding RNA that acts as a riboswitch and is folded into an appropriate stem-loop hairpin secondary structure with minimum free energy is predicted by several computational tools. Most of the secondary structures are similar but not the same, because of variation in sequence size. The TPP ligand can bind to the 3' untranslated region of the aptameric sequence, between the loops P2, P4, and P5 and the region between J2/3 and J4/5. The sequence of these loop regions in all predicted tertiary structure of the riboswitch in representative plant species--green algae to flowering plants--is the same, and the residues situated in these junctions are directly involved in binding thymine pyrophosphate and are conserved in all the representative species.

  6. Isopentenyl pyrophosphate is a novel antinociceptive substance that inhibits TRPV3 and TRPA1 ion channels.

    PubMed

    Bang, Sangsu; Yoo, Sungjae; Yang, Tae-Jin; Cho, Hawon; Hwang, Sun Wook

    2011-05-01

    Transient receptor potential ion channels (TRPs) expressed in the periphery sense and electrically transduce noxious stimuli to transmit the signals to the brain. Many natural and synthetic ligands for the sensory TRPs have been found, but little is known about endogenous inhibitors of these TRP channels. Recently, we reported that farnesyl pyrophosphate, an endogenous substance produced in the mevalonate pathway, is a specific activator for TRPV3. Here, we show that isopentenyl pyrophosphate (IPP), an upstream metabolite in the same pathway, is a dual inhibitor for TRPA1 and TRPV3. By using Ca(2+) imaging and voltage clamp experiments with human embryo kidney cell heterologous expression system, cultured sensory neurons, and epidermal keratinocytes, we demonstrate that micromolar IPP suppressed responses to specific agonists of TRPA1 and TRPV3. Consistently, peripheral IPP administration attenuated TRPA1 and TRPV3 agonist-specific acute pain behaviors. Furthermore, local IPP pretreatment significantly reversed mechanical and thermal hypersensitivity of inflamed animals. Taken together, the present study suggests that IPP is a novel endogenous TRPA1 and TRPV3 inhibitor that causes local antinociception. Our results may provide useful chemical information to elucidate TRP physiology in peripheral pain sensation.

  7. Mode of IMP and pyrophosphate enhancement of myosin and actin extraction from porcine meat.

    PubMed

    Nakamura, Yukinobu; Migita, Koshiro; Okitani, Akihiro; Matsuishi, Masanori

    2013-01-01

    We examined the mode of IMP and pyrophosphate enhancement of myosin and actin extraction from porcine meat. Extractabilities were determined after homogenates, prepared by adding 9 volumes of 0.3, 0.4, or 0.5 M NaCl solutions containing 0 to 36 mM IMP and 0 to 9 mM tetrapotassium pyrophosphate (KPP) to minced pork, were incubated at 4 °C for 0 or 12 h. Irrespective of the NaCl concentrations, IMP-induced extraction of both proteins increased with increasing extraction time. In contrast, that of KPP did not. When 0.3 M NaCl solutions containing both IMP and KPP were used, the solutions with 1.5 mM KPP showed marked enhancement of IMP-induced myosin and actin extraction. Incorporating these results with our previously published data (Nakamura et al., Biosci. Biotechnol. Biochem., 76, 1611-1615 (2012)), we hypothesized that IMP and KPP have the ability to release thick and thin filaments from restraints in myofibrils, in addition to the ability to dissociate actomyosin into myosin and actin, and that the restraint-releasing ability of IMP is dependent on reaction time and NaCl concentration while that of KPP is not.

  8. [Comparative effects of fluoride on three enzymes, hydrolyzing pyrophosphate - acid and alkaline phosphatases and inorganic pyrophosphatase].

    PubMed

    Kasho, V N; Baĭkov, A A; Avaeva, S M

    1982-08-01

    The effects of fluoride on the activities of acid phosphatase (EC 3.1.3.2) from potato and alkaline phosphatase (EC 3.1.3.1) from E. coli during pyrophosphate and p-nitrophenylphosphate hydrolysis and on the activities of inorganic pyrophosphatase (EC 3.6.1.1) from baker's yeast during pyrophosphate hydrolysis were compared. For both phosphatases the type of interaction was found to be independent on the nature of substrate. For acid phosphatase and inorganic pyrophosphatase the inhibition was of non-competitive and uncompetitive types, respectively. In the case of alkaline phosphatase fluoride increased the rate of p-nitrophenol release during p-nitrophenylphosphate hydrolysis at pH greater than or equal to 7.9 without affecting the rate of phosphate release, which is indicative of fluorophosphate formation in the course of the transphosphorylation reaction. The data obtained suggest the existence of essential differences in the mechanisms of fluoride effects on the three enzymes under study.

  9. Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Xanthomonas campestris pv. campestris.

    PubMed

    Frese, Marcel; Schatschneider, Sarah; Voss, Julia; Vorhölter, Frank-Jörg; Niehaus, Karsten

    2014-03-15

    Xanthomonads are plant pathogenic proteobacteria that produce the polysaccharide xanthan. They are assumed to catabolize glucose mainly via the Entner-Doudoroff pathway. Whereas previous studies have demonstrated no phosphofructokinase (PFK) activity in xanthomonads, detailed genome analysis revealed in Xanthomonas campestris pathovar campestris (Xcc) genes for all Embden-Meyerhof-Parnas pathway (glycolysis) enzymes, including a conserved pfkA gene similar to 6-phosphofructokinase genes. To address this discrepancy between genetic and physiological properties, the pfkA gene of Xcc strain B100 was cloned into the expression vector pET28a+. The 45-kDa pfkA gene product exhibited no conventional PFK activity. Bioinformatic analysis of the Xcc PfkA amino acid sequence suggested utilization of pyrophosphate as an alternative cosubstrate. Pyrophosphate-dependent PFK activity was shown in an in vitro enzyme assay for purified Xcc PfkA, as well as in the Xcc B100 crude protein extract. Kinetic constants were determined for the forward and reverse reactions. Primary structure conservation indicates the global presence of similar enzymes among Xanthomonadaceae.

  10. Calcium and bone disease

    PubMed Central

    Blair, Harry C.; Robinson, Lisa J.; Huang, Christopher L.-H.; Sun, Li; Friedman, Peter A.; Schlesinger, Paul H.; Zaidi, Mone

    2013-01-01

    Calcium transport and calcium signaling are of basic importance in bone cells. Bone is the major store of calcium and a key regulatory organ for calcium homeostasis. Bone, in major part, responds to calcium-dependent signals from the parathyroids and via vitamin D metabolites, although bone retains direct response to extracellular calcium if parathyroid regulation is lost. Improved understanding of calcium transporters and calcium-regulated cellular processes has resulted from analysis of genetic defects, including several defects with low or high bone mass. Osteoblasts deposit calcium by mechanisms including phosphate and calcium transport with alkalinization to absorb acid created by mineral deposition; cartilage calcium mineralization occurs by passive diffusion and phosphate production. Calcium mobilization by osteoclasts is mediated by acid secretion. Both bone forming and bone resorbing cells use calcium signals as regulators of differentiation and activity. This has been studied in more detail in osteoclasts, where both osteoclast differentiation and motility are regulated by calcium. PMID:21674636

  11. Calcium sulfate polymorph evolution dominated by competitive nucleation in gypsum metastable zone

    NASA Astrophysics Data System (ADS)

    Fu, Hailu; Jia, Caiyun; Chen, Qiaoshan; Jiang, Guangming

    2017-07-01

    Calcium sulfate polymorph evolution during spontaneous precipitation was probed in 1.75 m CaCl2 solutions at 85.0 °C. Metastable calcium sulfate dihydrate (DH) and unstable α-hemihydrate (α-HH) precipitated concomitantly with the dominant phase presenting an order of DH → α-HH → DH upon supersaturation. Competitive nucleation assisted with heterogeneous nucleation of relatively less stable phase at lower supersaturations accounts for such a polymorph evolution. Lower interfacial energy and higher supersaturation result in the DH formation, while heterogeneous nucleation triggers the α-HH formation. Increment in CaCl2 concentration favors α-HH precipitation by decreasing the water activity and narrowing the supersaturation gap between DH and α-HH. This work presents a facile rule manipulating the polymorph evolution and provides a control strategy for selective synthesis of α-HH in industry.

  12. Limitation of the antibiotic-eluting bone graft substitute: An example of gentamycin-impregnated calcium sulfate.

    PubMed

    Wu, Chang-Chin; Huang, Yang-Kai; Chang, Wei-Jen; Wu, Yun-Ching; Wang, Chen-Chie; Yang, Kai-Chiang

    2016-11-18

    Patients with inadequate volume of alveolar processes or bone defects commonly require graft substitutes in oral, maxillofacial or orthopedic surgery. Ridge augmentation and reconstruction of facial bony defects with bone graft materials achieve better outcomes in functional and aesthetic rehabilitation. The injectable calcium sulfate filler is used widely in intra-operative applications. Calcium sulfate bone filler has been shown to upregulate bone formation-related mRNA genes in vitro and improve osseointegration in vivo. In addition, the bone graft substitute can be used as a drug delivery system for antibiotics to treat or prevent infections based on the clinical experiences. However, the influences of antibiotics addition on the calcium sulfate are not fully understood. In this study, calcium sulfate impregnated with gentamycin in different weight ratios was characterized. The results showed that gentamycin prolonged the hydration process and extended initial/final setting times of calcium sulfate. The addition of gentamycin slowed the conversion from calcium sulfate hemihydrate to dihydrate and changed the crystalline phase and microstructure. Higher amounts of gentamycin added resulted in faster degradation and lower mechanical strength of calcium sulfate. This study reveals that the extended setting time, decreased compressive strength, and the accelerated degradation of the gentamycin-impregnated calcium sulfate bone graft substitutes should be considered during intra-operative applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2016. © 2016 Wiley Periodicals, Inc.

  13. Do Ca2+-adsorbing ceramics reduce the release of calcium ions from gypsum-based biomaterials?

    PubMed

    Belcarz, Anna; Zalewska, Justyna; Pałka, Krzysztof; Hajnos, Mieczysław; Ginalska, Grazyna

    2015-02-01

    Bone implantable materials based on calcium sulfate dihydrate dissolve quickly in tissue liquids and release calcium ions at very high levels. This phenomenon induces temporary toxicity for osteoblasts, may cause local inflammation and delay the healing process. Reduction in the calcium ion release rate by gypsum could be therefore beneficial for the healing of gypsum-filled bone defects. The aim of this study concerned the potential use of calcium phosphate ceramics of various porosities for the reduction of high Ca(2+) ion release from gypsum-based materials. Highly porous ceramics failed to reduce the level of Ca(2+) ions released to the medium in a continuous flow system. However, it succeeded to shorten the period of high calcium level. It was not the phase composition but the high porosity of ceramics that was found crucial for both the shortening of the Ca(2+) release-related toxicity period and intensification of apatite deposition on the composite. Nonporous ceramics was completely ineffective for this purpose and did not show any ability to absorb calcium ions at a significant level. Moreover, according to our observations, complex studies imitating in vivo systems, rather than standard tests, are essential for the proper evaluation of implantable biomaterials. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Transforming growth factor-beta1 incorporation in an alpha-tricalcium phosphate/dicalcium phosphate dihydrate/tetracalcium phosphate monoxide cement: release characteristics and physicochemical properties.

    PubMed

    Blom, E J; Klein-Nulend, J; Wolke, J G C; Kurashina, K; van Waas, M A J; Burger, E H

    2002-02-01

    The osteoconductive properties of calcium phosphate cements (CPCs) may be improved by the addition of growth factors, such as recombinant human transforming growth factor-beta1 (rhTGF-beta1). Previously we have shown that rhTGF-beta1 was released from cement enriched with rhTGF-beta1 and subsequently stimulated the differentiation of pre-osteoblastic cells from adult rat long bones. It is unknown whether the addition of rhTGF-beta1 changes the material properties of this alpha-tricalcium-phosphate (alpha-TCP)/tetracalcium-phosphate-monoxide (TeCP)/dicalcium-phosphate-dihydrate (DCPD) cement, and what the characteristics of the release of rhTGF-beta1 from this CPC are. Therefore, in the present study we determined the release of rhTGF-beta1 from cement pellets in vitro. The possible intervening effects of the CPC modification for intermixing rhTGF-beta1 on physicochemical properties were studied by assessing the compressive strength and setting time, as well as crystallinity, calcium to phosphorus ratio, porosity and microscopic structure. Most of the previously incorporated rhTGF-beta1 in the cement pellets was released within the first 48 h. For all concentrations of rhTGF-beta1 intermixed (100 ng-2.5 mg/g CPC), approximately 0.5% of the amount of rhTGF-beta1 incorporated initially was released in the first 2 h, increasing to 1.0% after 48 h. The release of rhTGF-beta1 continued hereafter at a rate of about 0.1% up to 1 week, after which no additional release was found. The initial setting time, nor the final setting time was changed in control cement without rhTGF-beta1 (standard CPC) or in cement modified for rhTGF-beta1 (modified CPC) at 20 degrees C and 37 degrees C. Setting times were more than six times decreased at 37 degrees C compared to 20 degrees C. The compressive strength was initially low for both standard CPC and modified CPC, after which it increased between 24 h and 8 weeks. The compressive strength for the modified CPC was significantly higher

  15. The influence of discharge power and heat treatment on calcium phosphate coatings prepared by RF magnetron sputtering deposition.

    PubMed

    Yonggang, Yan; Wolke, J G C; Yubao, Li; Jansen, J A

    2007-06-01

    Ca-P coatings with different Ca/P ratio and composition were successfully prepared by RF magnetron sputtering deposition. The Ca/P ratio, phase composition, structure and morphological properties were characterized by XRD, FTIR, EDS and SEM analyses. All the as-sputtered coatings were amorphous and after IR-irradiation the coatings altered into a crystalline phase. The obtained coatings had a Ca/P ratio that varied from 0.55 to 2.10 and different phase compositions or mixtures of apatite, beta-pyrophosphate and beta-tricalciumphosphate structures were formed. Evidently, the phase compositions of the sputtered coatings are determined not only by the discharge power ratio of the hydroxylapatite and calcium pyrophosphate targets but also by the annealing temperature.

  16. High-performance liquid chromatographic determination of pyrophosphate in the presence of a 20,000-fold excess of orthophosphate.

    PubMed

    Yoza, N; Akazaki, I; Nakazato, T; Ueda, N; Kodama, H; Tateda, A

    1991-12-01

    An HPLC method was based on anion-exchange separation of pyrophosphate (diphosphate) and orthophosphate and postcolumn spectrophotometric detection at 140 degrees C with a molybdenum(V)-molybdenum(VI) reagent. The reagent was easy to prepare, stable for at least 6 months at room temperature, and ready for the determination of pyrophosphate and orthophosphate by the so-called heteropoly blue method without use of any reducing agent. A photodiode-array detector for HPLC indicated the spectral characteristics of the heteropoply blue complex that was detectable at 330-800 nm. The HPLC method had a wide dynamic range from 3 x 10(-7) to 5 x 10(-4) M for both pyrophosphate and orthophosphate with a relative standard deviation of measurement of 10 approximately 2%. Pyrophosphate of 5 x 10(-7) and 5 x 10(-6) M, respectively, could be determined in the presence of a 20,000-fold excess of orthophosphate; 0.01 and 0.1 M.

  17. Structure of a Heterotetrameric Geranyl Pyrophosphate Synthase from Mint (Mentha piperita) Reveals Intersubunit Regulation[W][OA

    PubMed Central

    Chang, Tao-Hsin; Hsieh, Fu-Lien; Ko, Tzu-Ping; Teng, Kuo-Hsun; Liang, Po-Huang; Wang, Andrew H.-J.

    2010-01-01

    Terpenes (isoprenoids), derived from isoprenyl pyrophosphates, are versatile natural compounds that act as metabolism mediators, plant volatiles, and ecological communicators. Divergent evolution of homomeric prenyltransferases (PTSs) has allowed PTSs to optimize their active-site pockets to achieve catalytic fidelity and diversity. Little is known about heteromeric PTSs, particularly the mechanisms regulating formation of specific products. Here, we report the crystal structure of the (LSU · SSU)2-type (LSU/SSU = large/small subunit) heterotetrameric geranyl pyrophosphate synthase (GPPS) from mint (Mentha piperita). The LSU and SSU of mint GPPS are responsible for catalysis and regulation, respectively, and this SSU lacks the essential catalytic amino acid residues found in LSU and other PTSs. Whereas no activity was detected for individually expressed LSU or SSU, the intact (LSU · SSU)2 tetramer produced not only C10-GPP at the beginning of the reaction but also C20-GGPP (geranylgeranyl pyrophosphate) at longer reaction times. The activity for synthesizing C10-GPP and C20-GGPP, but not C15-farnesyl pyrophosphate, reflects a conserved active-site structure of the LSU and the closely related mustard (Sinapis alba) homodimeric GGPPS. Furthermore, using a genetic complementation system, we showed that no C20-GGPP is produced by the mint GPPS in vivo. Presumably through protein–protein interactions, the SSU remodels the active-site cavity of LSU for synthesizing C10-GPP, the precursor of volatile C10-monoterpenes. PMID:20139160

  18. Acetyl-CoA metabolism in amprolium-evoked thiamine pyrophosphate deficits in cholinergic SN56 neuroblastoma cells.

    PubMed

    Bizon-Zygmańska, D; Jankowska-Kulawy, A; Bielarczyk, H; Pawełczyk, T; Ronowska, A; Marszałł, M; Szutowicz, A

    2011-08-01

    Inhibition of pyruvate (PDHC) and ketoglutarate (KDHC) dehydrogenase complexes induced by thiamine pyrophosphate deficits is known cause of disturbances of cholinergic transmission in the brain, yielding clinical symptoms of cognitive, vegetative and motor deficits. However, particular alterations in distribution of key acetylcholine precursor, acetyl-CoA, in the cholinergic neuron compartment of thiamine pyrophosphate-deficient brain remain unknown. Therefore, the aim of our work was to find out how amprolium-induced thiamine pyrophosphate deficits (TD) affect distribution of acetyl-CoA in the compartment of pure cholinergic neuroblastoma SN56 cells originating from murine septum. Amprolium caused similar concentration-dependent decreases in thiamine pyrophosphate levels in nondifferentiated (NC) and differentiated (DC) cells cultured in low thiamine medium. In such conditions DC displayed significantly greater loss of viability than the NC ones, despite of lesser suppressions of PDHC activities and tetrazolium salt reduction rates in the former. On the other hand, intramitochondrial acetyl-CoA levels in DC were 73% lower than in NC, which explains their greater susceptibility to TD. Choline acetyltransferase activity and acetylcholine content in DC were two times higher than in NC. TD caused 50% decrease of cytoplasmic acetyl-CoA levels that correlated with losses of acetylcholine pool in DC but not in NC. These data indicate that particular sensitivity of DC to TD may result from relative shortage of acetyl-CoA due to its higher utilization in acetylcholine synthesis.

  19. A novel Y243S mutation in the pyruvate dehydrogenase El alpha gene subunit: correlation with thiamine pyrophosphate interaction.

    PubMed

    Benelli, C; Fouque, F; Redonnet-Vernhet, I; Malgat, M; Fontan, D; Marsac, C; Dey, R

    2002-08-01

    We identified a new Y243S mutation in the X-linked E1 alpha-PDH gene in a patient with pyruvate dehydrogenase complex (PDHc) deficiency. The activity in cultured fibroblasts was very low even in the presence of high thiamine pyrophosphate (TPP) concentrations, indicating that the defect could be due to decreased affinity of PDHc for TPP.

  20. Simple and rapid determination of hydrogen peroxide using phosphine-based fluorescent reagents with sodium tungstate dihydrate.

    PubMed

    Onoda, Maki; Uchiyama, Takefumi; Mawatari, Ken-Ichi; Kaneko, Kiyoko; Nakagomi, Kazuya

    2006-06-01

    A simple batch method for the fluorometric determination of hydrogen peroxide using phosphine-based fluorescent reagents has been developed. A rapid, mild and selective derivatization reaction was achieved by adding sodium tungstate dihydrate to the reaction mixture of hydrogen peroxide and a phosphine-based fluorescent reagent. When 4-diphenylphosphino-7-methylthio-2,1,3-benzoxadiazole was used as a reagent, the derivatization reaction was completed after 2 min at room temperature. The calibration curve was linear between 12.5 and 500 ng hydrogen peroxide in a 10 microL sample solution. This method is accurate and has potential for on-line applications.

  1. Active Calcium and Strontium Transport in Human Erythrocyte Ghosts

    PubMed Central

    Olson, Erik J.; Cazort, Ralph J.

    1969-01-01

    Both calcium and strontium could be transported actively from erythrocytes if adenosine triphosphate, guanosine triphosphate, or inosine triphosphate were included in the hypotonic medium used to infuse calcium or strontium into the cells. Acetyl phosphate and pyrophosphate were not energy sources for the transport of either ion. Neither calcium nor strontium transport was accompanied by magnesium exchange, and the addition of Mg++ to the reaction medium in a final concentration of 3.0 mmoles/liter did not promote the transport of either ion. In the absence of nucleotide triphosphates, the addition of 1.5 mmoles/liter of Sr++ to the reaction solution did not bring about active calcium transport and similarly 1.5 mmoles/liter of Ca++ did not bring about active strontium transport. The inclusion of 1.5 mmoles/liter of Ca++ or Sr++ in the reaction medium did not interfere with the transport of the other ion when the erythrocytes were infused with adenosine triphosphate. PMID:4304202

  2. Calcium and Vitamin D

    USDA-ARS?s Scientific Manuscript database

    Calcium is required for the bone formation phase of bone remodeling. Typically about 5 nmol (200 mg) of calcium is removed from the adult skeleton and replaced each day. To supply this amount, one would need to consume about 600 mg of calcium, since calcium is not very efficiently absorbed. Calcium ...

  3. Physicochemical characterization and pharmacokinetics in broiler chickens of a new recrystallized enrofloxacin hydrochloride dihydrate.

    PubMed

    Gutierrez, L; Miranda-Calderon, J E; Garcia-Gutierrez, P; Sumano, H

    2015-04-01

    Enrofloxacin, a key antimicrobial agent in commercial avian medicine, has limited bioavailability (60%). This prompted its chemical manipulation to yield a new solvate-recrystallized enrofloxacin hydrochloride dihydrate entity (enroC ). Its chemical structure was characterized by means of mass spectroscopy, Fourier transformed infrared spectroscopy, X-ray powder diffraction, and thermal analysis. Comparative oral pharmacokinetics (PK) of reference enrofloxacin (enroR ) and enroC in broiler chickens after oral administration revealed noticeable improvements in key parameters and PK/PD ratios. Maximum serum concentration values were 2.61 ± 0.21 and 5.9 ± 0.42 μg/mL for enroR and enroC , respectively; mean residence time was increased from 5.50 ± 0.26 h to 6.20 ± 0.71 h and the relative bioavailability of enroC was 336%. Considering Cmax /MIC and AUC/MIC ratios and the MIC values for a wild-type Escherichia coli O78/H12 (0.25 μg/mL), optimal ratios will only be achieved by enroC (Cmax /MIC = 23.6 and AUC/MIC = 197.7 for enroC ; vs. Cmax /MIC = 10.4 and AUC/MIC = 78.1 for enroR ). Furthermore, enroC may provide in most cases mutant prevention concentrations (Cmax /MIC ≥ 16). Ready solubility of powder enroC in drinking water at concentrations regularly used (0.01%) to provide an additional advantage of enroC in the field. Further development of enroC is warranted before it can be recommended for clinical use in veterinary medicine. © 2014 John Wiley & Sons Ltd.

  4. Cooperativity Assisted Shortening of Hydrogen Bonds in Crystalline Oxalic Acid Dihydrate: DFT and NBO Model Studies.

    PubMed

    Stare, Jernej; Hadži, Dušan

    2014-04-08

    The distance of ∼2.49 Å separating the carboxylic OH oxygen from the water oxygen atom in the α-polymorph of crystalline oxalic acid dihydrate is by ∼0.1 Å shorter than the average distance in carboxylic acid monohydrates. It is also by ∼0.2 Å shorter than the corresponding distance presently calculated for the heterotrimer consisting of one acid and two water molecules. The large difference between RO···O in the heterotrimer and in the crystal is attributed to the cooperative effect in the latter; this is supported by calculations carried out on clusters constituted of an increasing number of acid and water molecules. The present DFT calculations with geometry optimization include seven isolated model clusters, the largest of which contains five acid and eight water molecules. The RO···O of the short hydrogen bond shortens progressively with increasing the number of cluster constituents; in the largest cluster, it reaches 2.50 Å. This is remarkably close to both the experimental distance as well as to the distance obtained by the periodic DFT calculation. The electronic effects were studied by Natural Bond Orbital analysis, revealing an enhancement of hydrogen bonding on extending the network by increased polarization of the carbonyl group and by the increased delocalization interaction between the lone electron pair on the acceptor oxygen atom and the OH antibond orbital. The formation of circular motifs appears to be the most important factor in the cooperative shortening of the hydrogen bonds. In agreement with the measured hydrogen bond distances, inspection of the electron density reveals a notable difference in hydrogen bond shrinking tendency between the two known polymorphs of the title system.

  5. Spatial Distribution of Trehalose Dihydrate Crystallization in Tablets by X-ray Diffractometry.

    PubMed

    Thakral, Naveen K; Yamada, Hiroyuki; Stephenson, Gregory A; Suryanarayanan, Raj

    2015-10-05

    Crystallization of trehalose dihydrate (C12H22O11·2H2O) was induced by storing tablets of amorphous anhydrous trehalose (C12H22O11) at 65% RH (RT). Our goal was to evaluate the advantages and limitations of two approaches of profiling spatial distribution of drug crystallization in tablets. The extent of crystallization, as a function of depth, was determined in tablets stored for different time-periods. The first approach was glancing angle X-ray diffractometry, where the penetration depth of X-rays was modulated by the incident angle. Based on the mass attenuation coefficient of the matrix, the depth of X-ray penetration was calculated as a function of incident angle, which in turn enabled us to "calculate" the extent of crystallization to different depths. In the second approach, the tablets were split into halves and the split surfaces were analyzed directly. Starting from the tablet surface and moving toward the midplane, XRD patterns were collected in 36 "regions", in increments of 0.05 mm. The results obtained by the two approaches were, in general, in good agreement. Additionally, the results obtained were validated by determining the "average" crystallization in the entire tablet by using synchrotron radiation in the transmission mode. The glancing angle method could detect crystallization up to ∼650 μm and had a "surface bias". Being a nondestructive technique, this method will permit repeated analyses of the same tablet at different time points, for example, during a stability study. However, split tablet analyses, while a "destructive" technique, provided comprehensive and unbiased depth profiling information.

  6. Bovine albumin release and degradation analysis of dicalcium phosphate dihydrate cement.

    PubMed

    Metz, Jeremy; Sargent, Peter; Chu, Tien-Min G

    2006-01-01

    Dicalcium phosphate dihydrate (DCPD) cement was effective in our prior study as a bone morphogenetic protein-2 (BMP-2) delivery vehicle in a rat segmental defect regeneration study. In this study, we investigated the effects of liquid-to-powder (L/P) ratio on the in vitro degradation and protein release behavior of this material. The L/P ratios used in this study ranged from 0.50 to 0.83. DCPD cylinders were formed with a diameter of 1/4" and a height of 1/4". The effect of L/P on the initial compressive strength was found to be related to the porosity of the material at different L/P level. The strength of the material in phosphate buffered solution was found to degrade roughly 20% in 14 days. The relation between the final porosity and the compressive strength after degradation was modeled with Ryshkewitch equation. A liquid-to-powder ratio of 0.55, 0.7, and 0.8 was then used to fabricate samples for the protein release kinetic study. The low porosity (L/P = 0.55) group was found to have the fastest release rate, while the L/P = 0.8 group had the lowest. More then 60% of the loaded protein was released after 10 hours in all three groups with a final total release ranging between 75% and 93%. The findings suggested that the protein release profile of DCPD cements can be adjusted by the L/P ratio.

  7. Electron paramagnetic resonance spectroscopic study of copper hopping in doped bis(L-histidinato)cadmium dihydrate.

    PubMed

    Colaneri, Michael J; Vitali, Jacqueline; Kirschbaum, Kristin

    2013-04-25

    Electron paramagnetic resonance (EPR) spectroscopy was used to study Cu(II) dynamic behavior in a doped biological model crystal, bis(L-histidinato)cadmium dihydrate, in order to gain better insight into copper site stability in metalloproteins. Temperature-dependent changes in the low temperature X-band EPR spectra became visible around 100 K and continued up to room temperature. The measured 298 K g-tensor (principal values: 2.17, 2.16, 2.07) and copper hyperfine coupling tensor (principal values: -260, -190, -37 MHz) were similar to the average of the 77 K tensor values pertaining to two neighboring histidine binding sites. The observed temperature dependence was interpreted using Anderson's theory of motional narrowing, where the magnetic parameters for the different states are averaged as the copper rapidly hops between sites. The EPR pattern was also found to undergo a sharp sigmoidal-shaped, temperature-dependent conversion between two species with a critical temperature T(c) ≈ 160 K. The species below T(c) hops between the two low temperature site patterns, and the one above T(c) represents an average of the molecular spin Hamiltonian coupling tensors of the two 77 K sites. In addition, the low and high temperature species hop between one another, contributing to the dynamic averaging. Spectral simulations using this 4-state model determined a hop rate between the two low temperature sites ν(h4) = 4.5 × 10(8) s(-1) and between the low and high temperature states ν(h2) = 1.7 × 10(8) s(-1) at 160 K. An Arrhenius relationship of hop rate and temperature gave energy barriers of ΔE4 = 389 cm(-1) and ΔE2 = 656 cm(-1) between the two low temperature sites and between the low and high temperature states, respectively.

  8. ESR and optical study of Mn 2+-doped sodium hydrogen orthophosphate dihydrate

    NASA Astrophysics Data System (ADS)

    Kripal, Ram; Singh, D. K.

    2008-03-01

    ESR study of Mn 2+-doped sodium hydrogen orthophosphate dihydrate (SHOD) single crystals is done at room temperature. The Mn 2+ spin-Hamiltonian parameters have been evaluated employing a large number of resonant line positions observed for different orientations of the external magnetic field. The values of g, A, B, D, E and a are: 2.0042 ± 0.0002, 86 ± 2 × 10 -4 cm -1, 83 ± 2 × 10 -4 cm -1, 238 ± 2 × 10 -4 cm -1, 76 ± 2 × 10 -4 cm -1, 13 ± 1 × 10 -4 cm -1 for site I and 2.0032 ± 0.0002, 86 ± 2 × 10 -4 cm -1, 83 ± 2 × 10 -4 cm -1, 238 ± 2 × 10 -4 cm -1, 76 ± 2 × 10 -4 cm -1, 13 ± 1 × 10 -4 cm -1 for site II, respectively. The optical absorption study of the crystal is also done. The observed bands are assigned as transitions from the 6A 1g(S) ground state to various excited quartet levels of a Mn 2+ ion in a cubic crystalline field. These bands are fitted with four parameters B, C, Dq and α and the values found for the parameters are B = 777 cm -1, C = 3073 cm -1, Dq = 755 cm -1, and α = 76 cm -1. On the basis of the data obtained the surrounding crystalline field and the nature of metal-ligand bonding are discussed.

  9. Colorimetric sensing of pyrophosphate in aqueous media using bis-functionalised silica surfaces.

    PubMed

    Climent, Estela; Casasús, Rosa; Marcos, M Dolores; Martínez-Máñez, Ramón; Sancenón, Félix; Soto, Juan

    2009-06-28

    Bis-functionalised silica surfaces have been designed in order to develop selective and sensitive probes for the chromo-fluorogenic detection of certain guests. The designed system consists of a siliceous support bis-functinalised with thiol and polyamine groups. Thiol groups are suitable reactive centres (R) that are know to react with squaraine dyes (D) inducing bleaching, whereas polyamines act as host sites (H) able to coordinate certain target guests (G). In the absence of G, the reactive groups (R) react with the dye resulting in a bleaching of the solution. On the contrary, the presence of certain guest (G) results in a control of the reaction kinetic between R and D and eventually in a complete reaction inhibition. Different functionalised solids were prepared by reaction of the siliceous surface with different concentrations of mercaptopropyltrimethoxysilane (MPTS) and 3-[2-(2-aminoethylamino)ethylamino]propyl-trimethoxysilane (N3TS). The final materials (solids to ) were characterized employing standard procedures. In a first step the reactivity of the signaling dye D (squaraine ) with the reactive centre R (thiol groups) was studied as a function of the pH using solid that showed a most suitable response. At pH 7 and pH 5 there was a quick reaction between the squaraine and the thiol groups on the surface. On the contrary this reaction is significantly slower at pH 3 due to the different degree of protonation of the amines as a function of the pH. Additionally the reaction of with the squaraine has been studied in the presence of a range of inorganic anions with different structural dimensions and charges, including chloride, perchlorate, nitrate, sulfate, phosphate and pyrophosphate. At pH 3 the reaction of the dye with the thiol groups is still effective in the presence of chloride and perchlorate, but the reaction is highly inhibited in the presence of the anions nitrate, sulfate, phosphate and pyrophosphate. At pH 7 the squaraine reacts very fast

  10. The effects of 1% pyrophosphate and 0.02% sodium fluoride on artificial caries lesions in vivo.

    PubMed

    Mellberg, J R; Petrou, I D; Deutchman, M; Grote, N

    1988-12-01

    We evaluated the potential of a pyrophosphate-fluoride solution to affect the remineralizing-demineralizing equilibrium, i.e., caries-inhibiting/-promoting effects on enamel in vivo. Fifteen subjects carried dentin blocks and enamel thin sections with artificial caries lesions in removable partial dentures for periods of two weeks, during which time they rinsed twice daily in a double-blind, randomized cross-over design with solutions containing (a) 90 ppm F, (b) 90 ppm F and 1% pyrophosphate, or (c) no active agents (placebo). A severe cariogenic challenge provided to the lesions by plaque under a stainless steel mesh cover resulted in the placebo-treated lesions losing 70.2% +/- 72.1% mineral. The pyrophosphate rinse with fluoride held the mineral loss to only 28.1 +/- 52.8%, while the fluoride rinse without pyrophosphate held the loss to 24.2 +/- 50.1%. The differences between the fluoride and placebo results were significant (p less than 0.01), but the difference between the two fluoride groups was not. A large amount of fluoride was deposited in the dentin lesions. In the surface layer, the concentration was increased from 1000 ppm F to more than 2300 ppm F by both fluoride treatments. The concentration of fluoride in both groups of fluoride-treated lesions further increased to more than 3500 ppm F in the approximate center of the lesion before declining in deeper layers toward the level found in the placebo-treated group. The findings from both mineral change and fluoride uptake phases of this study show that in the presence of 90 ppm F, 1% pyrophosphate did not promote demineralization of artificial caries lesions.

  11. Calcium and Vitamin D

    MedlinePlus

    ... A calcium-rich diet (including dairy, nuts, leafy greens and fish) helps to build and protect your ... yogurt and cheese are high in calcium. Certain green vegetables and other foods contain calcium in smaller ...

  12. Molecular Cloning and Characterisation of Farnesyl Pyrophosphate Synthase from Tripterygium wilfordii

    PubMed Central

    Zhao, Yu-Jun; Chen, Xin; Zhang, Meng; Su, Ping; Liu, Yu-Jia; Tong, Yu-Ru; Wang, Xiu-Juan; Huang, Lu-Qi; Gao, Wei

    2015-01-01

    Farnesylpyrophosphate synthase (FPS) catalyzes the biosynthesis of farnesyl pyrophosphate (FPP), which is an important precursor of sesquiterpenoids such as artemisinin and wilfordine. In the present study, we report the molecular cloning and characterization of two full-length cDNAs encoding FPSs from Tripterygium wilfordii (TwFPSs). TwFPSs maintained their capability to synthesise FPP in vitro when purified as recombinant proteins from E. coli. Consistent with the endogenous role of FPS in FPP biosynthesis, TwFPSs were highly expressed in T. wilfordii roots, and were up-regulated upon methyl jasmonate (MeJA) treatment. The global gene expression profiles suggested that the TwFPSs might play an important regulatory role interpenoid biosynthesis in T. wilfordii, laying the groundwork for the future study of the synthetic biology of natural terpene products. PMID:25938487

  13. Colorimetric determination of pyrophosphate anion and its application to adenylation enzyme assay.

    PubMed

    Katano, Hajime; Watanabe, Hiro; Takakuwa, Masahiro; Maruyama, Chitose; Hamano, Yoshimitsu

    2013-01-01

    A colorimetric pyrophosphate assay based on the formation and reduction of the 18-molybdopyrophosphate ([(P2O7)Mo18O54](4-)) anion in an acetonitrile-water mixed solvent was modified and improved. The [(P2O7)Mo18O54](4-) anion is precipitated from the acetonitrile-water solution containing MoO4(2-) and HCl, and is re-dissolved in neat acetonitrile or propylene carbonate. This separation process decreases the interference by ATP, and prevents a yellow coloration of the reducing agent, ascorbic acid, due to excess Mo(VI) species. In the organic solvent, the [(P2O7)Mo18O54](4-) anion is reduced to a more intense blue molybdopyrophosphate species. The application of the colorimetry to the assay of adenylation enzymes is also described in this note.

  14. Nanomolar pyrophosphate detection and nucleus staining in living cells with simple terpyridine–Zn(II) complexes

    PubMed Central

    Chao, Duobin; Ni, Shitan

    2016-01-01

    Great efforts have been made to develop fluorescent probes for pyrophosphate (PPi) detection. Nucleus staining with fluorescence microscopy has been also widely investigated. But fluorescent probes for PPi detection with high sensitivity in water medium and nucleus staining with low–cost non–precious metal complexes in living cells are still challenging. Herein, we report simple terpyridine–Zn(II) complexes for selective nanomolar PPi detection over ATP and ADP in water based on aggregation induced emission (AIE) and intramolecular charge transfer (ICT). In addition, these terpyridine–Zn(II) complexes were successfully employed for nucleus staining in living cells. These results demonstrated simply obtained terpyridine–Zn(II) complexes are powerful tool for PPi detection and the development of PPi–related studies. PMID:27198968

  15. Pyrophosphate synthesis in iron mineral films and membranes simulating prebiotic submarine hydrothermal precipitates

    NASA Astrophysics Data System (ADS)

    Barge, Laura M.; Doloboff, Ivria J.; Russell, Michael J.; VanderVelde, David; White, Lauren M.; Stucky, Galen D.; Baum, Marc M.; Zeytounian, John; Kidd, Richard; Kanik, Isik

    2014-03-01

    Cells use three main ways of generating energy currency to drive metabolism: (i) conversion of adenosine diphosphate (ADP) to adenosine triphosphate (ATP) by the proton motive force through the rotor-stator ATP synthase; (ii) the synthesis of inorganic phosphate˜phosphate bonds via proton (or sodium) pyrophosphate synthase; or (iii) substrate-level phosphorylation through the direct donation from an active phosphoryl donor. A mechanism to produce a pyrophosphate bond as “energy currency” in prebiotic systems is one of the most important considerations for origin of life research. Baltscheffsky (1996) suggests that inorganic pyrophosphate (PO74-; PPi) may have preceded ATP/ADP as an energy storage molecule in earliest life, produced by an H+ pyrophosphatase. Here we test the hypothesis that PPi could be synthesized in inorganic precipitates simulating hydrothermal chimney structures transected by thermal and/or ionic gradients. Appreciable yields of PPi were obtained via substrate phosphorylation by acetyl phosphate within the iron sulfide/silicate precipitates at temperatures expected for an alkaline hydrothermal system. The formation of PPi only occurred in the solid phase, i.e. when both Pi and the phosphoryl donor were precipitated with Fe-sulfides or Fe-silicates. The amount of Ac-Pi incorporated into the precipitate was a significant factor in the amount of PPi that could form, and phosphate species were more effectively incorporated into the precipitate at higher temperatures (⩾50 to >85 °C). Thus, we expect that the hydrothermal precipitate would be more enriched in phosphate (and especially, Ac-Pi) near the inner margins of a hydrothermal mound where PPi formation would be at a maximum. Iron sulfide and iron silicate precipitates effectively stabilized Ac-Pi and PPi against hydrolysis (relative to hydrolysis in aqueous solution). Thus it is plausible that PPi could accumulate as an energy currency up to useful concentrations for early life in a

  16. Nanomolar pyrophosphate detection and nucleus staining in living cells with simple terpyridine–Zn(II) complexes

    NASA Astrophysics Data System (ADS)

    Chao, Duobin; Ni, Shitan

    2016-05-01

    Great efforts have been made to develop fluorescent probes for pyrophosphate (PPi) detection. Nucleus staining with fluorescence microscopy has been also widely investigated. But fluorescent probes for PPi detection with high sensitivity in water medium and nucleus staining with low–cost non–precious metal complexes in living cells are still challenging. Herein, we report simple terpyridine–Zn(II) complexes for selective nanomolar PPi detection over ATP and ADP in water based on aggregation induced emission (AIE) and intramolecular charge transfer (ICT). In addition, these terpyridine–Zn(II) complexes were successfully employed for nucleus staining in living cells. These results demonstrated simply obtained terpyridine–Zn(II) complexes are powerful tool for PPi detection and the development of PPi–related studies.

  17. Inositol pyrophosphate mediated pyrophosphorylation of AP3B1 regulates HIV-1 Gag release

    PubMed Central

    Azevedo, Cristina; Burton, Adam; Ruiz-Mateos, Ezequiel; Marsh, Mark; Saiardi, Adolfo

    2009-01-01

    High-energy inositol pyrophosphates, such as IP7 (diphosphoinositol pentakisphosphate), can directly donate a β-phosphate to a prephosphorylated serine residue generating pyrophosphorylated proteins. Here, we show that the β subunit of AP-3, a clathrin-associated protein complex required for HIV-1 release, is a target of IP7-mediated pyrophosphorylation. We have identified Kif3A, a motor protein of the kinesin superfamily, as an AP3B1-binding partner and demonstrate that Kif3A, like the AP-3 complex, is involved in an intracellular process required for HIV-1 Gag release. Importantly, IP7-mediated pyrophosphorylation of AP3B1 modulates the interaction with Kif3A and, as a consequence, affects the release of HIV-1 virus-like particles. This study identifies a cellular process that is regulated by IP7-mediated pyrophosphorylation. PMID:19934039

  18. Abdominal and hepatic uptake of /sup 99m/Tc-pyrophosphate in neonatal necrotizing enterocolitis

    SciTech Connect

    Caride, V.J.; Touloukian, R.J.; Ablow, R.C.; Lange, R.C.; Matthews, T.

    1981-04-01

    Abdominal /sup 99m/Tc-pyrophosphate (/sup 99m/Tc-PYP) scans were obtained in 15 neonates: 12 with neonatal necrotizing enterocolitis (NEC), two with osteomyelitis, and one with myocarditis. Ten of the babies with NEC had at least one positive scan; of these 10 studies, seven (Group A) showed both diffuse abdominal uptake and localized hepatic activity, two (Group B) showed abdominal uptake and questionable hepatic uptake, and one (Group C) demonstrated diffuse abdominal uptake only. The other two babies with NEC had normal scans (Group D). All NEC patients had normal scans. A patient with myocarditis had hepatic uptake of /sup 99m/Tc-PYP while the abdominal scan in the two infants with osteomyelitis was normal. These preliminary observations suggest that further study of a relationship between abdominal scan findings and the course of NEC is warranted.

  19. Thiamine pyrophosphate as an effector of 2-oxoglutarate dehydrogenase complex from European bison heart.

    PubMed

    Strumilo, S; Markiewicz, J

    1995-09-01

    The purified 2-oxoglutarate dehydrogenase complex (OGDC) from the European bison heart was near saturated with endogenous bound thiamine pyrophosphate (TPP). Exogenous TPP added to the full OGDC reaction medium decreased S0.5 for 2-oxoglutarate approximately 2.6-fold without any notable change in the maximum reaction rate. The TPP effect was observed in the presence of 1 mM ADP which alone is a strong positive allosteric effector of OGDC. At an unsaturating 2-oxoglutarate concentration the A50 value for TPP was approximately 0.05 mM. The ADP-like action of exogenous TPP was also found in the 2-oxoglutarate dehydrogenase (E1) reaction, determined in the presence of 2,6-dichlorophenoloindophenol as an electron acceptor.

  20. Identification and reconstitution of the yeast mitochondrial transporter for thiamine pyrophosphate.

    PubMed

    Marobbio, C M T; Vozza, A; Harding, M; Bisaccia, F; Palmieri, F; Walker, J E

    2002-11-01

    The genome of Saccharomyces cerevisiae contains 35 members of a family of transport proteins that, with a single exception, are found in the inner membranes of mitochondria. The transport functions of the 15 biochemically identified mitochondrial carriers are concerned with shuttling substrates, biosynthetic intermediates and cofactors across the inner membrane. Here the identification of the mitochondrial carrier for the essential cofactor thiamine pyrophosphate (ThPP) is described. The protein has been overexpressed in bacteria, reconstituted into phospholipid vesicles and identified by its transport properties. In confirmation of its identity, cells lacking the gene for this carrier had reduced levels of ThPP in their mitochondria, and decreased activity of acetolactate synthase, a ThPP-requiring enzyme found in the organellar matrix. They also required thiamine for growth on fermentative carbon sources.

  1. Nano-conjugate fluorescence probe for the discrimination of phosphate and pyrophosphate.

    PubMed

    Kim, Ik-Bum; Han, Man H; Phillips, Ronnie L; Samanta, Bappaditya; Rotello, Vincent M; Zhang, Z John; Bunz, Uwe H F

    2009-01-01

    We describe a pyrophosphate (PPi) probe that is based on a fluorescent dicarboxylate-substituted poly(para-phenyleneethynylene) (PPE) and 10 nm cobalt-iron spinel nanoparticles (NPs) in aqueous media. The spinel NPs efficiently quench the fluorescence of the PPE at a concentration of 20-30 pmol. Addition of phosphate anions to the PPE-NP construct displaces the quenched PPE to give rise to a fluorescent response; we found that PPi and phosphate (Pi) have significantly different binding affinities for the self-assembled materials. We can discern >40 nM PPi in the presence of 0.1 mM Pi at pH 7, which suggests that these assemblies may be useful in bio-analytical applications. This displacement assay was used to effectively determine the ability of pyrophosphatase to hydrolyze PPi to Pi.

  2. Role of arginine in the binding of thiamin pyrophosphate to Escherichia coli pyruvate oxidase.

    PubMed

    Koland, J G; O'Brien, T A; Gennis, R B

    1982-05-25

    The mode of interaction between Escherichia coli pyruvate oxidase and its cofactor, thiamin pyrophosphate (TPP), was studied with the aid of arginine-directed reagents. The enzyme is rapidly inactivated by either phenylglyoxal or 2,3-butanedione, with the cofactor, TPP, offering partial protection against these reagents. The inactivation by phenylglyoxal was found to be reversible. Experiments with [7-14C]phenylglyoxal showed that while several arginine residues react with this reagent, TPP can prevent the labeling of one such residue. Furthermore, inactivation by 2,3-butanedione is attended by at least a 100-fold decrease in affinity of the enzyme for TPP. These results suggest a direct role for arginine in the binding of the cofactor.

  3. New lithium iron pyrophosphate as 3.5 V class cathode material for lithium ion battery.

    PubMed

    Nishimura, Shin-ichi; Nakamura, Megumi; Natsui, Ryuichi; Yamada, Atsuo

    2010-10-06

    A new pyrophosphate compound Li(2)FeP(2)O(7) was synthesized by a conventional solid-state reaction, and its crystal structure was determined. Its reversible electrode operation at ca. 3.5 V vs Li was identified with the capacity of a one-electron theoretical value of 110 mAh g(-1) even for ca. 1 μm particles without any special efforts such as nanosizing or carbon coating. Li(2)FeP(2)O(7) and its derivatives should provide a new platform for related lithium battery electrode research and could be potential competitors to commercial olivine LiFePO(4), which has been recognized as the most promising positive cathode for a lithium-ion battery system for large-scale applications, such as plug-in hybrid electric vehicles.

  4. A novel role for farnesyl pyrophosphate synthase in fibroblast growth factor-mediated signal transduction.

    PubMed Central

    Reilly, John F; Martinez, Shawndra D; Mickey, Gregory; Maher, Pamela A

    2002-01-01

    Farnesyl pyrophosphate synthase (FPPS) catalyses the formation of a key cellular intermediate in isoprenoid metabolic pathways. Here we describe a novel role for this enzyme in fibroblast growth factor (FGF)-mediated signalling. We demonstrate the binding of FPPS to FGF receptors (FGFRs) using the yeast two-hybrid assay, pull-down assays and co-immunoprecipitation. The interaction between FPPS and FGFR is regulated by the cellular metabolic state and by treatment with FGF-2. Overexpression of FPPS inhibits FGF-2-induced cell proliferation, accompanied by a failure of the FGF-2-mediated induction of cyclins D1 and E. Overexpression of FPPS in fibroblasts also promotes increased farnesylation of Ras, and temporally extends FGF-2-stimulated activation of the Ras/ERK (extracellular-signal-regulated kinase) cascade. These data suggest that, in addition to its role in isoprenoid biosynthesis, FPPS may function as a modulator of the cellular response to FGF treatment. PMID:12020352

  5. Inositol pyrophosphates inhibit Akt signaling, regulate insulin sensitivity and weight gain

    PubMed Central

    Chakraborty, Anutosh; Koldobskiy, Michael A.; Bello, Nicholas T.; Maxwell, Micah; Potter, James J.; Juluri, Krishna R.; Maag, David; Kim, Seyun; Huang, Alex S.; Dailey, Megan J.; Saleh, Masoumeh; Snowman, Adele M.; Moran, Timothy H.; Mezey, Esteban; Snyder, Solomon H.

    2011-01-01

    Summary The inositol pyrophosphate IP7 (5-diphosphoinositolpentakisphosphate), formed by a family of three inositol hexakisphosphate kinases (IP6Ks), modulates diverse cellular activities. We now report that IP7 is a physiologic inhibitor of Akt, a serine/threonine kinase which regulates glucose homeostasis and protein translation respectively via the GSK3β and mTOR pathways. Thus Akt, mTOR and GSK3β signaling are dramatically augmented in skeletal muscle, white adipose tissue, and liver of mice with targeted deletion of IP6K1. IP7 impacts this pathway by potently inhibiting the PDK1 phosphorylation of Akt, preventing its activation and thereby impacting insulin signaling. IP6K1 knockout mice manifest insulin sensitivity and are resistant to obesity elicited by high fat diet or aging. Inhibition of IP6K1 may afford a therapeutic approach to obesity and diabetes. PMID:21145457

  6. Live-Cell Pyrophosphate Imaging by in Situ Hot-Spot Generation.

    PubMed

    Li, Mingmin; Li, Jin; Di, Huixia; Liu, Huiqiao; Liu, Dingbin

    2017-03-21

    Controlling the electromagnetic hot-spot generation is essential for surface-enhanced Raman scattering (SERS) assays. Current hot-spot-based SERS assays have been extensively studied in solutions or on substrates. However, probing biospecies by controlling the hot-spot assembly in living systems has not been demonstrated thus far. Herein, we report a background-free SERS probe for imaging pyrophosphate (PPi), a biochemically significant anion, in living cells. Intracellular PPi is able to induce the nanoparticle dimerization, thus creating an intense electromagnetic hot spot and dramatically enhancing the signal of the Raman reporters residing in the hot spot. More impressively, the reporter we used in this study provides a strong and sharp single peak in the cellular Raman-silent region (1800-2800 cm(-1)), thus eliminating the possible background interference. This strategy could be readily extended to detect other biomarkers by only replacing the recognition ligands.

  7. Molecular Cloning and Characterisation of Farnesyl Pyrophosphate Synthase from Tripterygium wilfordii.

    PubMed

    Zhao, Yu-Jun; Chen, Xin; Zhang, Meng; Su, Ping; Liu, Yu-Jia; Tong, Yu-Ru; Wang, Xiu-Juan; Huang, Lu-Qi; Gao, Wei

    2015-01-01

    Farnesylpyrophosphate synthase (FPS) catalyzes the biosynthesis of farnesyl pyrophosphate (FPP), which is an important precursor of sesquiterpenoids such as artemisinin and wilfordine. In the present study, we report the molecular cloning and characterization of two full-length cDNAs encoding FPSs from Tripterygium wilfordii (TwFPSs). TwFPSs maintained their capability to synthesise FPP in vitro when purified as recombinant proteins from E. coli. Consistent with the endogenous role of FPS in FPP biosynthesis, TwFPSs were highly expressed in T. wilfordii roots, and were up-regulated upon methyl jasmonate (MeJA) treatment. The global gene expression profiles suggested that the TwFPSs might play an important regulatory role interpenoid biosynthesis in T. wilfordii, laying the groundwork for the future study of the synthetic biology of natural terpene products.

  8. Determination of Single-Nucleotide Polymorphisms by Real-time Pyrophosphate DNA Sequencing

    PubMed Central

    Alderborn, Anders; Kristofferson, Anna; Hammerling, Ulf

    2000-01-01

    The characterization of naturally occurring variations in the human genome has evoked an immense interest during recent years. Variations known as biallelic Single-Nucleotide Polymorphisms (SNPs) have become increasingly popular markers in molecular genetics because of their wide application both in evolutionary relationship studies and in the identification of susceptibility to common diseases. We have addressed the issue of SNP genotype determination by investigating variations within the Renin–Angiotensin–Aldosterone System (RAAS) using pyrosequencing, a real-time pyrophosphate detection technology. The method is based on indirect luminometric quantification of the pyrophosphate that is released as a result of nucleotide incorporation onto an amplified template. The technical platform employed comprises a highly automated sequencing instrument that allows the analysis of 96 samples within 10 to 20 minutes. In addition to each studied polymorphic position, 5–10 downstream bases were sequenced for acquisition of reference signals. Evaluation of pyrogram data was accomplished by comparison of peak heights, which are proportional to the number of incorporated nucleotides. Analysis of the pyrograms that resulted from alternate allelic configurations for each addressed SNP revealed a highly discriminating pattern. Homozygous samples produced clear-cut single base peaks in the expected position, whereas heterozygous counterparts were characterized by distinct half-height peaks representing both allelic positions. Whenever any of the allelic bases of an SNP formed a homopolymer with adjacent bases, the nonallelic signal was added to those of the SNP. This feature did not, however, influence SNP readability. Furthermore, the multibase reading capacity of the described system provides extensive flexibility in regard to the positioning of sequencing primers and allows the determination of several closely located SNPs in a single run. PMID:10958643

  9. Fungal Inositol Pyrophosphate IP7 Is Crucial for Metabolic Adaptation to the Host Environment and Pathogenicity

    PubMed Central

    Lev, Sophie; Li, Cecilia; Desmarini, Desmarini; Saiardi, Adolfo; Fewings, Nicole L.; Schibeci, Stephen D.; Sharma, Raghwa; Sorrell, Tania C.

    2015-01-01

    ABSTRACT Inositol pyrophosphates (PP-IPs) comprising inositol, phosphate, and pyrophosphate (PP) are essential for multiple functions in eukaryotes. Their role in fungal pathogens has never been addressed. Cryptococcus neoformans is a model pathogenic fungus causing life-threatening meningoencephalitis. We investigate the cryptococcal kinases responsible for the production of PP-IPs (IP7/IP8) and the hierarchy of PP-IP importance in pathogenicity. Using gene deletion and inositol polyphosphate profiling, we identified Kcs1 as the major IP6 kinase (producing IP7) and Asp1 as an IP7 kinase (producing IP8). We show that Kcs1-derived IP7 is the most crucial PP-IP for cryptococcal drug susceptibility and the production of virulence determinants. In particular, Kcs1 kinase activity is essential for cryptococcal infection of mouse lungs, as reduced fungal burdens were observed in the absence of Kcs1 or when Kcs1 was catalytically inactive. Transcriptome and carbon source utilization analysis suggested that compromised growth of the KCS1 deletion strain (Δkcs1 mutant) in the low-glucose environment of the host lung is due to its inability to utilize alternative carbon sources. Despite this metabolic defect, the Δkcs1 mutant established persistent, low-level asymptomatic pulmonary infection but failed to elicit a strong immune response in vivo and in vitro and was not readily phagocytosed by primary or immortalized monocytes. Reduced recognition of the Δkcs1 cells by monocytes correlated with reduced exposure of mannoproteins on the Δkcs1 mutant cell surface. We conclude that IP7 is essential for fungal metabolic adaptation to the host environment, immune recognition, and pathogenicity. PMID:26037119

  10. Iron status and food matrix strongly affect the relative bioavailability of ferric pyrophosphate in humans.

    PubMed

    Moretti, Diego; Zimmermann, Michael B; Wegmüller, Rita; Walczyk, Thomas; Zeder, Christophe; Hurrell, Richard F

    2006-03-01

    Although ferric pyrophosphate is a promising compound for iron fortification of foods, few data are available on the effect of food matrices, processing, and ascorbic acid on its bioavailability. We compared the relative bioavailability (RBV) of ferrous sulfate in an experimental form of micronized dispersible ferric pyrophosphate (MDFP) in a wheat-milk infant cereal given with and without ascorbic acid with the RBV of MDFP from a processed and unprocessed rice meal. A crossover design was used to measure iron absorption in young women (n = 26) from test meals fortified with isotopically labeled [57Fe]-MDFP and [58Fe]-ferrous sulfate, based on erythrocyte incorporation of stable isotope labels 14 d later. Geometric mean iron absorption from the wheat-based meal fortified with MDFP was 2.0% and that from the meal fortified with ferrous sulfate was 3.2% (RBV = 62). The addition of ascorbic acid at a molar ratio of 4:1 to iron increased iron absorption from MDFP to 5.8% and that from ferrous sulfate to 14.8% (RBV = 39). In the rice meals, mean iron absorption from MDFP added to the rice at the time of feeding was 1.7%, and that from ferrous sulfate was 11.6% (RBV = 15). The mean iron absorption from MDFP extruded into artificial rice grains was 3.0% and that from ferrous sulfate in unprocessed rice was 12.6% (RBV = 24). Sixteen of 26 subjects were iron deficient. Iron status was a highly significant predictor of the RBV of MDFP (P < 0.001). RBV of the experimental MDFP varied markedly with food matrix and iron status. Assigning a single RBV value to poorly soluble compounds may be of limited value in evaluating their suitability for food fortification.

  11. Direct observation of multiple tautomers of oxythiamine and their recognition by the thiamine pyrophosphate riboswitch.

    PubMed

    Singh, Vipender; Peng, Chunte Sam; Li, Deyu; Mitra, Koyel; Silvestre, Katherine J; Tokmakoff, Andrei; Essigmann, John M

    2014-01-17

    Structural diversification of canonical nucleic acid bases and nucleotide analogues by tautomerism has been proposed to be a powerful on/off switching mechanism allowing regulation of many biological processes mediated by RNA enzymes and aptamers. Despite the suspected biological importance of tautomerism, attempts to observe minor tautomeric forms in nucleic acid or hybrid nucleic acid-ligand complexes have met with challenges due to the lack of sensitive methods. Here, a combination of spectroscopic, biochemical, and computational tools probed tautomerism in the context of an RNA aptamer-ligand complex; studies involved a model ligand, oxythiamine pyrophosphate (OxyTPP), bound to the thiamine pyrophosphate (TPP) riboswitch (an RNA aptamer) as well as its unbound nonphosphorylated form, oxythiamine (OxyT). OxyTPP, similarly to canonical heteroaromatic nucleic acid bases, has a pyrimidine ring that forms hydrogen bonding interactions with the riboswitch. Tautomerism was established using two-dimensional infrared (2D IR) spectroscopy, variable temperature FTIR and NMR spectroscopies, binding isotope effects (BIEs), and computational methods. All three possible tautomers of OxyT, including the minor enol tautomer, were directly identified, and their distributions were quantitated. In the bound form, BIE data suggested that OxyTPP existed as a 4'-keto tautomer that was likely protonated at the N1'-position. These results also provide a mechanistic framework for understanding the activation of riboswitch in response to deamination of the active form of vitamin B1 (or TPP). The combination of methods reported here revealing the fine details of tautomerism can be applied to other systems where the importance of tautomerism is suspected.

  12. Technical problems associated with the production of technetium Tc 99m tin(II) pyrophosphate kits

    SciTech Connect

    Kowalsky, R.J.; Dalton, D.R.

    1981-11-01

    The amount of tin(II) required for adequate reduction, complexation, and stability of technetium Tc 99m pertechnetate in radiopharmaceutical kits, and methods of preventing the loss of tin(II) during formulation of these lyophilized kits are investigated. Tin(II) loss from stannous chloride solutions was studied under several conditions, including room air versus nitrogen atmospheres, during vial filling in a laminar-flow hood with samples frozen on dry ice versus samples at room temperature, during lyophilization, and during storage under refrigerated, ambient, and elevated temperatures. Various amounts of stannous chloride, ranging from 5 to 1000 microgram/ml, were used in formulating sodium pertechnetate Tc 99m kits containing 100 mCi technetium Tc 99m and 0.4 microgram total technetium. Samples were removed at various times; hydrolyzed technetium, pertechnetate, and technetium Tc 99m pyrophosphate were isolated on instant thin-layer chromatography-silica gel and quantified with a scintillation counter. The time necessary to deoxygenate distilled water by nitrogen purging was measured. Several sources of stannous chloride were assayed for tin(II) content. Tin(II) loss occurs rapidly in solution (15% in one hour) unless continuously protected with nitrogen, and during vial filling in a laminar-flow hood unless frozen with dry ice. No substantial loss of tin(II) was detected during lyophilization or during storage of lyophilized product at any of the three temperatures. A minimum of 400 microgram tin(II) was required to provide 90% technetium Tc 99m pyrophosphate at six hours after preparation. Adequate deoxygenation of small quantities (450 ml) of water was accomplished in less than one hour. Some stannous chloride salts were highly oxidized in the dry state, and only high-purity elemental tin wire gave acceptable yields of tin(II).

  13. Technical problems associated with the production of technetium Tc 99m tin(II) pyrophosphate kits.

    PubMed

    Kowalsky, R J; Dalton, D R

    1981-11-01

    The amount of tin(II) required for adequate reduction, complexation, and stability of technetium Tc 99m pertechnetate in radiopharmaceutical kits, and methods of preventing the loss of tin(II) during formulation of these lyophilized kits are investigated. Tin(II) loss from stannous chloride solutions was studied under several conditions, including room air versus nitrogen atmospheres, during vial filling in a laminar-flow hood with samples frozen on dry ice versus samples at room temperature, during lyophilization, and during storage under refrigerated, ambient, and elevated temperatures. Various amounts of stannous chloride, ranging from 5 to 1000 microgram/ml, were used in formulating sodium pertechnetate Tc 99m kits containing 100 mCi technetium Tc 99m and 0.4 microgram total technetium. Samples were removed at various times; hydrolyzed technetium, pertechnetate, and technetium Tc 99m pyrophosphate were isolated on instant thin-layer chromatography-silica gel and quantified with a scintillation counter. The time necessary to deoxygenate distilled water by nitrogen purging was measured. Several sources of stannous chloride were assayed for tin(II) content. Tin(II) loss occurs rapidly in solution (15% in one hour) unless continuously protected with nitrogen, and during vial filling in a laminar-flow hood unless frozen with dry ice. No substantial loss of tin(II) was detected during lyophilization or during storage of lyophilized product at any of the three temperatures. A minimum of 400 microgram tin(II) was required to provide 90% technetium Tc 99m pyrophosphate at six hours after preparation. Adequate deoxygenation of small quantities (450 ml) of water was accomplished in less than one hour. Some stannous chloride salts were highly oxidized in the dry state, and only high-purity elemental tin wire gave acceptable yields of tin(II).

  14. Chemical modulation of crystalline state of calcium oxalate with nickel ions.

    PubMed

    Akhtar, Khalida; Haq, Ikram Ul

    2013-03-15

    We explored that the presence of nickel ions in the precipitation medium affected size, shape and crystalline phase of the precipitated particles of calcium oxalate, whereas the applied synthesis conditions strongly influenced their uniformity. Aqueous solutions of oxalic acid and calcium chloride, containing varying amounts of nickel sulfate, were mixed at room temperature and allowed to sonicate for various periods of time. The obtained particles were characterized by SEM, XRD, TG/DTA, and FTIR. Results revealed that particle morphology of calcium oxalate and their hydration states were dependent upon the chemical composition of the reactant solutions. For instance, the particles precipitated out in the form of dihydrate and having prismatic, and discs shaped particle morphology, when nickel ions were introduced in the starting reactant solution in different amounts. In contrast, the calcium oxalate particles precipitated under identical conditions in the absence of nickel ions were in the form of flakes with corrugated edges. Obvious variations were also found in the XRD patterns and crystallite size of these three solids. Heat treatment produced changes in the surface morphology of these particles due to loss of material and converted them calcium oxide. Gentle mixing of aqueous solutions of calcium chloride and oxalic acid in the absence and presence of nickel ions produced precipitated particles of calcium oxalate. Chemical composition of the reactant solutions and their order of mixing were found to be the key parameters in controlling the particle morphology and their hydration/crystalline state. Heat treatment at the elevated temperature transformed the as-prepared calcium oxalate to calcium oxide with visible changes in surface features of the particles. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Effect of experimental and sample factors on dehydration kinetics of mildronate dihydrate: mechanism of dehydration and determination of kinetic parameters.

    PubMed

    Bērziņš, Agris; Actiņš, Andris

    2014-06-01

    The dehydration kinetics of mildronate dihydrate [3-(1,1,1-trimethylhydrazin-1-ium-2-yl)propionate dihydrate] was analyzed in isothermal and nonisothermal modes. The particle size, sample preparation and storage, sample weight, nitrogen flow rate, relative humidity, and sample history were varied in order to evaluate the effect of these factors and to more accurately interpret the data obtained from such analysis. It was determined that comparable kinetic parameters can be obtained in both isothermal and nonisothermal mode. However, dehydration activation energy values obtained in nonisothermal mode showed variation with conversion degree because of different rate-limiting step energy at higher temperature. Moreover, carrying out experiments in this mode required consideration of additional experimental complications. Our study of the different sample and experimental factor effect revealed information about changes of the dehydration rate-limiting step energy, variable contribution from different rate limiting steps, as well as clarified the dehydration mechanism. Procedures for convenient and fast determination of dehydration kinetic parameters were offered.

  16. Influence of shot peening on corrosion properties of biocompatible magnesium alloy AZ31 coated by dicalcium phosphate dihydrate (DCPD).

    PubMed

    Mhaede, Mansour; Pastorek, Filip; Hadzima, Branislav

    2014-06-01

    Magnesium alloys are promising materials for biomedical applications because of many outstanding properties like biodegradation, bioactivity and their specific density and Young's modulus are closer to bone than the commonly used metallic implant materials. Unfortunately their fatigue properties and low corrosion resistance negatively influenced their application possibilities in the field of biomedicine. These problems could be diminished through appropriate surface treatments. This study evaluates the influence of a surface pre-treatment by shot peening and shot peening+coating on the corrosion properties of magnesium alloy AZ31. The dicalcium phosphate dihydrate coating (DCPD) was electrochemically deposited in a solution containing 0.1M Ca(NO3)2, 0.06M NH4H2PO4 and 10mL/L of H2O2. The effect of shot peening on the surface properties of magnesium alloy was evaluated by microhardness and surface roughness measurements. The influence of the shot peening and dicalcium phosphate dihydrate layer on the electrochemical characteristics of AZ31 magnesium alloy was evaluated by potentiodynamic measurements and electrochemical impedance spectroscopy in 0.9% NaCl solution at a temperature of 22±1°C. The obtained results were analyzed by the Tafel-extrapolation method and equivalent circuit method. The results showed that the application of shot peening process followed by DCPD coating improves the properties of the AZ31 surface from corrosion and mechanical point of view.

  17. Study of N, N-dimethyl(carboethoxymethyl)-3-phthalimidopropylammonium chloride dihydrate by DFT calculations, NMR and FTIR spectroscopy

    NASA Astrophysics Data System (ADS)

    Kowalczyk, Iwona

    2009-06-01

    N, N-dimethyl(carboethoxymethyl)-3-phthalimidopropylammonium chloride dihydrate ( 1) and N, N-dimethyl(carboxymethyl)-3-phthalimidopropylammonium hydrochloride ( 3) have been obtained in reaction of N, N-dimethyl-3-phthalimidopropylamine with ethyl chloroacetate and chloroacetic acid, respectively. N, N-dimethyl(carboethoxymethyl)-3-phthalimido-propylammonium chloride dihydrate ( 1) has been characterized by FTIR and NMR spectroscopy. Moreover, for ( 1) and ( 3) B3LYP calculations have been carried out. The optimized bond lengths, bond angles and torsion angles calculated by B3LYP/6-31G(d,p) approach have been presented. Both FTIR and Raman spectra of ( 1) are consistent with the calculated structures in the gas phase. Correlations between the experimental 1H and 13C NMR chemical shifts ( δexp) of investigated compound in D 2O, and the GIAO/B3LYP/6-31G(d,p) calculated magnetic isotropic shielding tensors ( σcalc), δexp = a + b σcalc, are reported. The assignments of the anharmonic experimental solid state vibrational frequencies of ( 1) based on the calculated B3LYP/6-31G(d,p) harmonic frequencies have been made. Linear correlations between the experimental 1H and 13C chemical shifts and the computed screening constants confirm the optimized geometry.

  18. Morphological control of calcium oxalate particles in the presence of poly-(styrene-alt-maleic acid)

    NASA Astrophysics Data System (ADS)

    Yu, Jiaguo; Tang, Hua; Cheng, Bei; Zhao, Xiujian

    2004-10-01

    Calcium oxalate (CaOx) particles exhibiting different shapes and phase structures were fabricated by a simple precipitation reaction of sodium oxalate with calcium chloride in the absence and presence of poly-(styrene-alt-maleic acid) (PSMA) as a crystal modifier at room temperature. The as-obtained products were characterized with scanning electron microscopy (SEM) and X-ray diffraction (XRD). The effects of reaction conditions including pH, [Ca2+]/[C2O42-] ratio and concentration of PSMA and CaC2O4 on the crystal forms and morphologies of the as-obtained calcium oxalate were investigated. The results show that various crystal morphologies of calcium oxalate, such as parallelograms, plates, spheres, bipyramids etc. can be obtained depending on the experimental conditions. Higher polymer concentration favors formation of the metastable calcium oxalate dihydrate (COD) crystals. Lower pH is beneficial to the formation of plate-like CaOx crystals. Especially, the monodispersed parallelogram-like CaOx crystals can be produced by PSMA as an additive at pH 2. PSMA may act as a good inhibitor for urolithiasis since it induces the formation of COD and reduces the particle size of CaOx. This research may provide new insight into the morphological control of CaOx particles and the prevention of urolithiasis.

  19. Thallium-201/technetium-99m pyrophosphate overlap in patients with acute myocardial infarction after thrombolysis: prediction of depressed wall motion despite thallium uptake

    SciTech Connect

    Schofer, J.; Spielmann, R.P.; Broemel, T.B.; Bleifeld, W.; Mathey, D.G.

    1986-08-01

    Intracoronary thallium-201/technetium-99m pyrophosphate planar scintigraphy was performed in 60 patients with acute myocardial infarction undergoing intracoronary thrombolysis to predict salvage of myocardium immediately after thrombolysis. In eight patients a significant overlap of new thallium uptake and technetium pyrophosphate accumulation was found after thrombolysis. Intravenous planar thallium scintigraphy revealed thallium uptake in the region of overlap in all patients; circumferential profile analysis showed no difference in the thallium scintigrams before and after technetium injections. Both findings indicate that overlap is not the result of scattering of technetium into the thallium window. Emission computed tomography revealed thallium/technetium pyrophosphate uptake in identical slices and regions. Regional wall motion in the area of overlap remained depressed in all patients, in contrast to patients with similar thallium uptake without overlap. These data suggest that thallium/technetium pyrophosphate overlap reflects the close proximity of viable and necrotic myocardial cells and predicts depressed wall motion after thrombolysis.

  20. Efficacy of a new recrystallized enrofloxacin hydrochloride-dihydrate against leptospirosis in a hamster model.

    PubMed

    Carrascosa, Alma; Gutierrez, Lilia; De la Peña, Alejandro; Candanosa, Irma E; Tapia, Graciela; Sumano, Hector

    2017-09-05

    A trial on Syrian hamsters (Mesocricetus auratus) infected with Leptospira interrogans serovar Canicola was established to compare treatment efficacies of daily IM injections of either 10 mg/kg of Baytril® 5% or the same dose of enrofloxacin dihydrate-hydrochloride (enro-C). Hamsters were experimentally infected via oral submucosa with 400 microorganisms/animal, in a sequential time-schedule to align initial treatment day as follows: group BE24 treated with Baytril® daily for 7 days after 24 of infection; group enro-C24 treated with enro-C as in group BE24; group BE74 treated also with Baytril®, but starting 72 h after infection; group enro-C74 treated as group BE74 but injecting enro-C. An untreated-uninfected control group (CG-) and an infected-untreated control group (CG+) were assembled (n =18 in all groups). Hamsters' weights and temperatures were monitored daily for 28 days. Once euthanatized or following death, necropsy, histopathology, macroscopic agglutination tests (MAT), bacterial culture and PCR were performed. Mortality was 38.8% in group BE24 and 100 % in group BE74 No mortality was observed in enro-C24 and 11.1% mortality was recorded in enro-C74 Mortality in groups CG+ and CG- was 100% and zero, respectively. Combined necropsy and histopathologic findings revealed signs of septicemia and organ damage in groups BE24, BE72 and CG(+) Groups enro-C24 and and CG-showed no lesions. Moderated lesions were registered in 3 hamsters in group enro-C72 MAT was positive in 83.3 % of BE24 hamsters (83.3 %) and 100 % in BE72 and CG+; 16.7% in Enro-C24 and 38.9 % in enro-C72 Only 4/18 were PCR positive in enro-C72 and only one in enro-C24 (P<0.05). It can be concluded that enro-C may be a viable option to treat leptospirosis in hamsters and that this may be the case in other species. Copyright © 2017 American Society for Microbiology.

  1. Calcium and Calcium-Base Alloys

    DTIC Science & Technology

    1949-01-01

    alloys have •been made in electrical contacts. Little is known of’ the high - calcium alloys,» The aluminum-calcium diagram from Hansen^1) is shown in...list is still incom- plete« No use has been suggested for high calcium -aluminum alloys, ..•Arsenic-pal’c-iüm- Alloys •K.. Calcium arsenide, OajAsg...hot CaCUy, by X-ray determination of the structure. The probability of finding a useful high - calcium alloy in this system is based-on-the-validity

  2. Raman spectroscopy study of calcium oxalate extracted from cacti stems.

    PubMed

    Frausto-Reyes, Claudio; Loza-Cornejo, Sofia; Terrazas, Teresa; Terrazas, Tania; Miranda-Beltrán, María de la Luz; Aparicio-Fernández, Xóchitl; López-Macías, Brenda M; Morales-Martínez, Sandra E; Ortiz-Morales, Martín

    2014-01-01

    To find markers that distinguish the different Cactaceae species, by using near infrared Raman spectroscopy and scanning electron microscopy, we studied the occurrence, in the stem, of solid deposits in five Cactaceae species (Coryphantha clavata, Ferocactus latispinus, Opuntia ficus-indica, O. robusta, and O. strepthacantha) collected from their natural habitats from a region of México. The deposits in the tissues usually occurred as spheroidal aggregates, druses, or prismatic crystals. From the Raman spectra, the crystals were identified either as calcium oxalate monohydrate (CaC2O4·H2O) or calcium oxalate dihydrate (CaC2O4·2H2O). Opuntia species (subfamily Opuntioideae) showed the presence of CaC2O4·H2O, and the deposition of CaC2O4·2H2O was present in C. clavata and F. latispinus (subfamily Cactoideae, Cacteae tribe). As a punctual technique, Raman spectroscopy seems to be a useful tool to identify crystal composition. In addition to allowing the analysis of crystal morphology, this spectroscopic technique can be used to identify Cactaceae species and their chemotaxonomy.

  3. Metastable states in calcium phosphate - aqueous phase equilibrations

    NASA Astrophysics Data System (ADS)

    Driessens, F. C. M.; Verbeeck, R. M. H.

    1981-05-01

    A critical evaluation of the literature reveals that during equilibration of well crystallized hydroxyapatite in aqueous solutions metastable states can occur. They are characterized by a persistent supersaturation with respect to hydroxyapatite and a systematical dependence of the ion activity product of this compound on the solution composition. For products synthesized by thermal treatment it is known that they are transformed into oxyhydroxyapatite so that the theoretical solubility behaviour could be predicted from the extrapolated value of the free energy of oxyapatite at room temperature: the negative logarithm of the ionic product for hydroxyapatite should become close to that of oxyapatite during equilibration. The discrepancy with experimental data is probably due to the formation of thin layers seeming dicalcium phosphate dihydrate, octocalcium phosphate or defective hydroxyapatite as coatings on the apatite crystals. This is derived from the apparent Ca/P ratio of the solubility controlling phase. According to chemical potential plots this apparent Ca/P ratio can have values close to 1, 1.33, 1.50 or 1.67. The aqueous solutions are clearly undersaturated with respect to the more acidic calcium phosphates so that the coatings must deviate from the compositions of these compounds in their pure state. The formation of these metastable states during equilibration of oxyhydroxyapatites is compared with others occuring during precipitation and crystal growth of calcium phosphates. A model is proposed which explains the observations qualitatively.

  4. A simple colorimetric assay method for pyrophosphate in the presence of a 1000-fold excess of orthophosphate: application of the method to the study of pyrophosphate metabolism in mitochondria.

    PubMed

    Mansurova, S E; Deryabina, O A

    1989-02-01

    A sensitive colorimetric method for the assay of inorganic pyrophosphate with excess of orthophosphate is described. The principle of this method lies in the formation of phosphomolybdate and PPi-molybdate complexes with subsequent extraction of the phosphomolybdate complex by organic solvents and reduction of the PPi-molybdate complex by dithiothreitol and Eikonogen. The sensitivity of the method was from 5 to 120 nmol of PPi in a 2.0-ml sample.

  5. Postoperative myocardial infarction documented by technetium pyrophosphate scan using single-photon emission computed tomography: Significance of intraoperative myocardial ischemia and hemodynamic control

    SciTech Connect

    Cheng, D.C.; Chung, F.; Burns, R.J.; Houston, P.L.; Feindel, C.M. )

    1989-12-01

    The aim of this prospective study was to document postoperative myocardial infarction (PMI) by technetium pyrophosphate scan using single-photon emission computed tomography (TcPPi-SPECT) in 28 patients undergoing elective coronary bypass grafting (CABG). The relationships of intraoperative electrocardiographic myocardial ischemia, hemodynamic responses, and pharmacological requirements to this incidence of PMI were correlated. Radionuclide cardioangiography and TcPPi-SPECT were performed 24 h preoperatively and 48 h postoperatively. A standard high-dose fentanyl anesthetic protocol was used. Twenty-five percent of elective CABG patients were complicated with PMI, as documented by TcPPi-SPECT with an infarcted mass of 38.0 +/- 5.5 g. No significant difference in demographic, preoperative right and left ventricular function, number of coronary vessels grafted, or aortic cross-clamp time was observed between the PMI and non-PMI groups. The distribution of patients using preoperative beta-adrenergic blocking drugs or calcium channel blocking drugs was found to have no correlation with the outcome of PMI. As well, no significant differences in hemodynamic changes or pharmacological requirements were observed in the PMI and non-PMI groups during prebypass or postbypass periods, indicating careful intraoperative control of hemodynamic indices did not prevent the outcome of PMI in these patients. However, the incidence of prebypass ischemia was 39.3% and significantly correlated with the outcome of positive TcPPi-SPECT, denoting a 3.9-fold increased risk of developing PMI. Prebypass ischemic changes in leads II and V5 were shown to correlate with increased CPK-MB release (P less than 0.05) and tends to occur more frequently with lateral myocardial infarction.

  6. Inorganic phosphate (Pi) modulates the expression of key regulatory proteins of the inorganic pyrophosphate (PPi) metabolism in TGF-β1-stimulated chondrocytes.

    PubMed

    Hamade, Tala; Bianchi, Arnaud; Sebillaud, Sylvie; Netter, Patrick; Jouzeau, Jean-Yves; Cailotto, Frédéric

    2010-01-01

    The balance between extracellular inorganic phosphate (ePi) and extracellular inorganic pyrophosphate (ePPi) is controlled by four membrane proteins: the transporters ANK (exporting PPi outside the cells) and PiT-1 (importing ePi into the cells), and the enzymes PC-1 (generating ePPi from nucleotides) and Tissue Non-specific Alkaline Phosphatase (TNAP, hydrolyzing ePPi into ePi). TGF-β1 was shown to stimulate ANK and PC-1 expression in articular chondrocytes, and subsequent ePPi level, as well as to increase ePi uptake by inducing PiT-1 expression in a chondrogenic cell line. Thus, we investigated the ability of ePi to modulate the effect of TGF-β1 on the regulatory proteins of the ePi/ePPi balance in chondrocytes. In the pathophysiological range of 0.01-1 mM, ePi was inactive by itself but potentiated the stimulatory effects of TGF-β1 on ANK, PC-1 or PiT-1 mRNA (RT-qPCR) and protein (Western blot) levels. PC-1 activity was also increased by TGF-β1 and further potentiated by ePi supplementation. TNAP mRNA and activity became undetectable in response to TGF-β1. These data suggest that ePi could increase ePPi level by changing the control of ANK and PC-1 expression by TGF-β1, further highlighting an adaptative regulation of the Pi/PPi balance to prevent basic calcium phosphate deposition into the joints.

  7. A Novel Inositol Pyrophosphate Phosphatase in Saccharomyces cerevisiae: Siw14 PROTEIN SELECTIVELY CLEAVES THE β-PHOSPHATE FROM 5-DIPHOSPHOINOSITOL PENTAKISPHOSPHATE (5PP-IP5).

    PubMed

    Steidle, Elizabeth A; Chong, Lucy S; Wu, Mingxuan; Crooke, Elliott; Fiedler, Dorothea; Resnick, Adam C; Rolfes, Ronda J

    2016-03-25

    Inositol pyrophosphates are high energy signaling molecules involved in cellular processes, such as energetic metabolism, telomere maintenance, stress responses, and vesicle trafficking, and can mediate protein phosphorylation. Although the inositol kinases underlying inositol pyrophosphate biosynthesis are well characterized, the phosphatases that selectively regulate their cellular pools are not fully described. The diphosphoinositol phosphate phosphohydrolase enzymes of the Nudix protein family have been demonstrated to dephosphorylate inositol pyrophosphates; however, theSaccharomyces cerevisiaehomolog Ddp1 prefers inorganic polyphosphate over inositol pyrophosphates. We identified a novel phosphatase of the recently discovered atypical dual specificity phosphatase family as a physiological inositol pyrophosphate phosphatase. Purified recombinant Siw14 hydrolyzes the β-phosphate from 5-diphosphoinositol pentakisphosphate (5PP-IP5or IP7)in vitro. In vivo,siw14Δ yeast mutants possess increased IP7levels, whereas heterologousSIW14overexpression eliminates IP7from cells. IP7levels increased proportionately whensiw14Δ was combined withddp1Δ orvip1Δ, indicating independent activity by the enzymes encoded by these genes. We conclude that Siw14 is a physiological phosphatase that modulates inositol pyrophosphate metabolism by dephosphorylating the IP7isoform 5PP-IP5to IP6.

  8. An investigation of the effect of thiamine pyrophosphate on cisplatin-induced oxidative stress and DNA damage in rat brain tissue compared with thiamine: thiamine and thiamine pyrophosphate effects on cisplatin neurotoxicity.

    PubMed

    Turan, M I; Cayir, A; Cetin, N; Suleyman, H; Siltelioglu Turan, I; Tan, H

    2014-01-01

    This study investigated the effects of thiamine pyrophosphate (TPP) at dosages of 10 and 20 mg/kg on oxidative stress induced in rat brain tissue with cisplatin and compared this with thiamine. Cisplatin neurotoxicity represents one of the main restrictions on the drug being given in effective doses. Oxidative stress is considered responsible for cisplatin toxicity. Our results showed that cisplatin increased the levels of oxidant parameters such as lipid peroxidation (thio barbituric acid reactive substance (TBARS)) and myeloperoxidase (MPO) in brain tissue and suppressed the effects of antioxidants such as total glutathione (GSH) and superoxide dismutase (SOD). TPP, especially at a dosage of 20 mg/kg, significantly reduced TBARS and MPO levels that increase with cisplatin administration compared with the thiamine group, while TPP significantly increases GSH and SOD levels. In addition, the level of 8-Gua (guanine), a product of DNA damage, was 1.7 ± 0.12 8-hydroxyl guanine (8-OH Gua)/105 Gua in brain tissue in the control group receiving cisplatin, compared with 0.97 ± 0.03 8-OH Gua/105 Gua in the thiamine pyrophosphate (20 mg/kg) group and 1.55 ± 0.11 8-OH Gua/105 Gua in the thiamine (20 mg/kg) group. These results show that thiamine pyrophosphate significantly prevents oxidative damage induced by cisplatin in brain tissue, while the protective effect of thiamine is insignificant.

  9. Fabrication of calcium phosphate–calcium sulfate injectable bone substitute using hydroxy-propyl-methyl-cellulose and citric acid

    PubMed Central

    Thai, Van Viet

    2010-01-01

    In this study, an injectable bone substitute (IBS) consisting of citric acid, chitosan, and hydroxyl propyl methyl cellulose (HPMC) as the liquid phase and tetra calcium phosphate (TTCP), dicalcium phosphate dihydrate (DCPD) and calcium sulfate dehydrate (CSD, CaSO4·2H2O) powders as the solid phase, were fabricated. Two groups were classified based on the percent of citric acid in the liquid phase (20, 40 wt%). In each groups, the HPMC percentage was 0, 2, and 4 wt%. An increase in compressive strength due to changes in morphology was confirmed by scanning electron microscopy images. A good conversion rate of HAp at 20% citric acid was observed in the XRD profiles. In addition, HPMC was not obviously affected by apatite formation. However, both HPMC and citric acid increased the compressive strength of IBS. The maximum compressive strength for IBS was with 40% citric acid and 4% HPMC after 14 days of incubation in 100% humidity at 37°C. PMID:20333539

  10. Fabrication of calcium phosphate-calcium sulfate injectable bone substitute using hydroxy-propyl-methyl-cellulose and citric acid.

    PubMed

    Thai, Van Viet; Lee, Byong-Taek

    2010-06-01

    In this study, an injectable bone substitute (IBS) consisting of citric acid, chitosan, and hydroxyl propyl methyl cellulose (HPMC) as the liquid phase and tetra calcium phosphate (TTCP), dicalcium phosphate dihydrate (DCPD) and calcium sulfate dehydrate (CSD, CaSO4 x 2H2O) powders as the solid phase, were fabricated. Two groups were classified based on the percent of citric acid in the liquid phase (20, 40 wt%). In each groups, the HPMC percentage was 0, 2, and 4 wt%. An increase in compressive strength due to changes in morphology was confirmed by scanning electron microscopy images. A good conversion rate of HAp at 20% citric acid was observed in the XRD profiles. In addition, HPMC was not obviously affected by apatite formation. However, both HPMC and citric acid increased the compressive strength of IBS. The maximum compressive strength for IBS was with 40% citric acid and 4% HPMC after 14 days of incubation in 100% humidity at 37 degrees C.

  11. Imaging calcium in neurons.

    PubMed

    Grienberger, Christine; Konnerth, Arthur

    2012-03-08

    Calcium ions generate versatile intracellular signals that control key functions in all types of neurons. Imaging calcium in neurons is particularly important because calcium signals exert their highly specific functions in well-defined cellular subcompartments. In this Primer, we briefly review the general mechanisms of neuronal calcium signaling. We then introduce the calcium imaging devices, including confocal and two-photon microscopy as well as miniaturized devices that are used in freely moving animals. We provide an overview of the classical chemical fluorescent calcium indicators and of the protein-based genetically encoded calcium indicators. Using application examples, we introduce new developments in the field, such as calcium imaging in awake, behaving animals and the use of calcium imaging for mapping single spine sensory inputs in cortical neurons in vivo. We conclude by providing an outlook on the prospects of calcium imaging for the analysis of neuronal signaling and plasticity in various animal models.

  12. Label-free electrical detection of pyrophosphate generated from DNA polymerase reactions on field-effect devices†

    PubMed Central

    Su, Xing; Wu, Kai; Elibol, Oguz H.; Liu, David J.; Reddy, Bobby; Tsai, Ta-Wei; Dorvel, Brian R.; Daniels, Jonathan S.

    2012-01-01

    We introduce a label-free approach for sensing polymerase reactions on deoxyribonucleic acid (DNA) using a chelator-modified silicon-on-insulator field-effect transistor (SOI-FET) that exhibits selective and reversible electrical response to pyrophosphate anions. The chemical modification of the sensor surface was designed to include rolling-circle amplification (RCA) DNA colonies for locally enhanced pyrophosphate (PPi) signal generation and sensors with immobilized chelators for capture and surface-sensitive detection of diffusible reaction by-products. While detecting arrays of enzymatic base incorporation reactions is typically accomplished using optical fluorescence or chemiluminescence techniques, our results suggest that it is possible to develop scalable and portable PPi-specific sensors and platforms for broad biomedical applications such as DNA sequencing and microbe detection using surface-sensitive electrical readout techniques. PMID:22262038

  13. Three-dimensional structure of phosphoribosyl pyrophosphate synthetase from E. coli at 2.71 Å resolution

    NASA Astrophysics Data System (ADS)

    Timofeev, V. I.; Abramchik, Yu. A.; Zhukhlistova, N. E.; Muravieva, T. I.; Esipov, R. S.; Kuranova, I. P.

    2016-01-01

    Phosphoribosyl pyrophosphate synthetase from Escherichia coli was cloned, purified, and crystallized. Single crystals of the enzyme were grown under microgravity. The X-ray diffraction data set was collected at the Spring-8 synchrotron facility and used to determine the three-dimensional structure of the enzyme by the molecular-replacement method at 2.71 Å resolution. The active and regulatory sites in the molecule of E. coli phosphoribosyl pyrophosphate synthetase were revealed by comparison with the homologous protein from Bacillus subtilis, the structure of which was determined in a complex with functional ligands. The conformations of polypeptide-chain fragments surrounding and composing the active and regulatory sites were shown to be identical in both proteins.

  14. The use of /sup 99/Tc-pyrophosphate for imaging and measuring of calcification caused by implanted polymers

    SciTech Connect

    Cifkova, I.; Sprincl, L.

    1980-11-01

    Pyrophosphate labeled with /sup 99/Tc was evaluated as an indicator of the calcification which occurs after implantation of polymeric materials in rats. Poly(2-hydroxyethyl methacrylate), both porous and homogeneous, had been implanted. /sup 99/Tc pyrophosphate was applied intravenously to rats in intervals from 14 days to 15 months. Scintigraphy, x-ray examination, and histology were carried out. The ratio of specific activity in the capsule around the implant to specific activity in the reference tissue was determined. The cumulation of activity was parallel with the development of calcification as revealed by histology. The method can be used not only in the screening test of new polymeric materials, but also in a quantitative determination of the degree of calcification in general.

  15. Label-free electrical detection of pyrophosphate generated from DNA polymerase reactions on field-effect devices.

    PubMed

    Credo, Grace M; Su, Xing; Wu, Kai; Elibol, Oguz H; Liu, David J; Reddy, Bobby; Tsai, Ta-Wei; Dorvel, Brian R; Daniels, Jonathan S; Bashir, Rashid; Varma, Madoo

    2012-03-21

    We introduce a label-free approach for sensing polymerase reactions on deoxyribonucleic acid (DNA) using a chelator-modified silicon-on-insulator field-effect transistor (SOI-FET) that exhibits selective and reversible electrical response to pyrophosphate anions. The chemical modification of the sensor surface was designed to include rolling-circle amplification (RCA) DNA colonies for locally enhanced pyrophosphate (PPi) signal generation and sensors with immobilized chelators for capture and surface-sensitive detection of diffusible reaction by-products. While detecting arrays of enzymatic base incorporation reactions is typically accomplished using optical fluorescence or chemiluminescence techniques, our results suggest that it is possible to develop scalable and portable PPi-specific sensors and platforms for broad biomedical applications such as DNA sequencing and microbe detection using surface-sensitive electrical readout techniques.

  16. Three-dimensional structure of phosphoribosyl pyrophosphate synthetase from E. coli at 2.71 Å resolution

    SciTech Connect

    Timofeev, V. I. E-mail: tostars@mail.ru; Abramchik, Yu. A.; Zhukhlistova, N. E.; Muravieva, T. I.; Esipov, R. S.; Kuranova, I. P.

    2016-01-15

    Phosphoribosyl pyrophosphate synthetase from Escherichia coli was cloned, purified, and crystallized. Single crystals of the enzyme were grown under microgravity. The X-ray diffraction data set was collected at the Spring-8 synchrotron facility and used to determine the three-dimensional structure of the enzyme by the molecular-replacement method at 2.71 Å resolution. The active and regulatory sites in the molecule of E. coli phosphoribosyl pyrophosphate synthetase were revealed by comparison with the homologous protein from Bacillus subtilis, the structure of which was determined in a complex with functional ligands. The conformations of polypeptide-chain fragments surrounding and composing the active and regulatory sites were shown to be identical in both proteins.

  17. Magnetite nanoparticle-induced fluorescence quenching of adenosine triphosphate-BODIPY Conjugates: application to adenosine triphosphate and pyrophosphate sensing.

    PubMed

    Yu, Cheng-Ju; Wu, Su-Mei; Tseng, Wei-Lung

    2013-09-17

    We report that magnetite nanoparticles (Fe3O4 NPs) act as an efficient quencher for boron dipyrromethene-conjugated adenosine 5'-triphosphate (BODIPY-ATP) that is highly fluorescent in bulk solution. BODIPY-ATP molecules attached to the surface of Fe3O4 NPs through the coordination between the triphosphate group of BODIPY-ATP and Fe(3+)/Fe(2+) on the NP surface. The formed complexes induced an apparent reduction in the BODIPY-ATP fluorescence resulting from an oxidative-photoinduced electron transfer (PET) from the BODIPY-ATP excited state to an unfilled d shell of Fe(3+)/Fe(2+) on the NP surface. A comparison of the Stern-Volmer quenching constant between Fe(3+) and Fe(2+) suggests that Fe(3+) on the NP surface dominantly controls this quenching process. The efficiency for Fe3O4 NP-induced fluorescence quenching of the BODIPY-ATP was enhanced by increasing the concentration of Fe3O4 NPs and lowering the pH of the solution to below 6.0. We found that pyrophosphate and ATP compete with BODIPY-ATP for binding to Fe3O4 NPs. Thus, we amplified BODIPY-ATP fluorescence in the presence of increasing the pyrophosphate and ATP concentration; the detection limits at a signal-to-noise ratio of 3 for pyrophosphate and ATP were determined to be 7 and 30 nM, respectively. The Fe3O4 NP-based competitive binding assay detected ATP and pyrophosphate in only 5 min. The selectivity of this assay for ATP over metal ions, amino acids, and adenosine analogues is particularly high. The practicality of using the developed method to determine ATP in a single drop of blood is also validated.

  18. An Evaluative History of Bisphosphonate Drugs: Dual Physiologic Effects of Pyrophosphate as Inspiration for a Novel Pharmaceutical Class

    PubMed Central

    2016-01-01

    The documented history of the development of the bisphosphonate drugs is reviewed in sufficient detail to permit independent evaluation of the consistency of the conclusions reached from the available data. The evidence developed during the early interval of these studies 1960–1975 was sufficient to establish that pyrophosphate shares the subsequently established dual bisphosphonate characteristics of bone resorption inhibition and prevention of tissue mineralization. PMID:27800209

  19. Chemical and biological reduction of Mn (III)-pyrophosphate complexes: Potential importance of dissolved Mn (III) as an environmental oxidant

    NASA Astrophysics Data System (ADS)

    Kostka, Joel E.; Luther, George W., III; Nealson, Kenneth H.

    1995-03-01

    Dissolved Mn (III) is a strong oxidant which could play an important role in the biogeochemistry of aquatic environments, but little is known about this form of Mn. Mn(III) was shown to form a stable complex with pyrophosphate which is easily measured by uv-vis spectrophotometry. The Mn(III)-pyrophosphate complex was produced at concentrations of 5 μM to 10 mM Mn at neutral pH. Inorganic electron donors, Fe(II) and sulfide, abiotically reduced Mn(III)-pyrophosphate in seconds with a stoichiometry of 1:1 and near 1:2 reductant:Mn (III), respectively. Shewanella putrefaciens strain MR-1 catalyzed the reduction of Mn(III)-pyrophosphate with formate or lactate as electron donors. Reduction of Mn(III) catalyzed by MR-1 was inhibited under aerobic conditions but only slightly under anaerobic conditions upon addition of the alternate electron acceptor, nitrate. MR-1 catalyzed reduction was also inhibited by metabolic inhibitors including formaldehyde, tetrachlorosalicylanilide (TCS), carbonyl cyanide m-chlorophenylhydrazone (CCCP), 2- n-heptyl-4-hydroxyquinoline N-oxide (HQNO), but not antimycin A. When formate or lactate served as electron donor for Mn(III) reduction, carbon oxidation to CO 2 was coupled to the respiration of Mn (III). Using the incorporation of 3H-leucine into the TCA-insoluble fraction of culture extracts, it was shown that Mn (III) reduction was coupled to protein synthesis in MR-1. These data indicate that Mn (III) complexes may be produced under conditions found in aquatic environments and that the reduction of Mn(III) can be coupled to the cycling of Fe, S, and C.

  20. Radionuclide imaging in myocardial sarcoidosis. Demonstration of myocardial uptake of /sup 99m/Tc pyrophosphate and gallium

    SciTech Connect

    Forman, M.B.; Sandler, M.P.; Sacks, G.A.; Kronenberg, M.W.; Powers, T.A.

    1983-03-01

    A patient had severe congestive cardiomyopathy secondary to myocardial sarcoidosis. The clinical diagnosis was confirmed by radionuclide ventriculography, /sup 201/Tl, /sup 67/Ga, and /sup 99m/Tc pyrophosphate (TcPYP) scintigraphy. Myocardial TcPYP uptake has not been reported previously in sarcoidosis. In this patient, TcPYP was as useful as gallium scanning and thallium imaging in documenting the myocardial process.