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Sample records for 32p 203hg 198au

  1. Determination of 198Au X-rays emission probabilities.

    PubMed

    Moreira, D S; Koskinas, M F; Dias, M S; Yamazaki, I M

    2010-01-01

    This work describes the measurements of the K X-ray and gamma-ray emission probabilities per decay of (198)Au performed at the Nuclear Metrology Laboratory (LMN) at the IPEN, São Paulo. The radioactive sample was obtained by means of (197)Au(n, gamma)(198)Au reaction irradiating an Au foil in a thermal neutron flux near the core of the IPEN 3.5 MW research reactor. The activity of samples was determined in a 4pibeta-gamma coincidence system, setting the gamma window at the 411.80 keV total energy absorption peak. The same samples were measured in two different spectrometers: a HPGe planar spectrometer with Be window, suitable for measurements in the low energy range and a coaxial REGe spectrometer. Both spectrometers were previously calibrated in a well defined geometry by means of standard sources calibrated in a 4pibeta-gamma coincidence system. MCNP4C Monte Carlo code was used for simulating the REGe spectrometer calibration curve, and a new version of code ESQUEMA was adopted for simulating the detection processes in the coincidence system, in order to predict the efficiency extrapolation curve. Copyright 2009 Elsevier Ltd. All rights reserved.

  2. Dosimetry of the 198Au Source used in Interstitial Brachytherapy

    SciTech Connect

    Dauffy, L; Braby, L; Berner, B

    2004-05-18

    The American Association of Physicists in Medicine Task Group 43 report, AAPM TG-43, provides an analytical model and a dosimetry protocol for brachytherapy dose calculations, as well as documentation and results for some sealed sources. The radionuclide {sup 198}Au (T{sub 1/2} = 2.70 days, E{gamma} = 412 keV) has been used in the form of seeds for brachytherapy treatments including brain, eye, and prostate tumors. However, the TG-43 report has no data for {sup 198}Au seeds, and none have previously been obtained. For that reason, and because of the conversion of most treatment planning systems to TG-43 based methods, both Monte Carlo calculations (MCNP 4C) and thermoluminescent dosimeters (TLDs) are used in this work to determine these data. The geometric variation in dose is measured using an array of TLDs in a solid water phantom, and the seed activity is determined using both a well ion chamber and a High Purity Germanium detector (HPGe). The results for air kerma strength, S{sub k}, per unit apparent activity, are 2.06 (MCNP) and 2.09 (measured) U mCi{sup -1}. The former is identical to what was published in 1991 in the AAPM Task Group 32 report. The dose rate constant results, {Lambda}, are 1.12 (MCNP) and 1.10 (measured), cGy h{sup -1} U{sup -1}. The radial dose function, g(r), anisotropy function, F(r,{theta}), and anisotropy factor, {psi}{sub an}(r), are given. The anisotropy constant values are 0.973 (MCNP) and 0.994 (measured) and are consistent with both source geometry and the emitted photon energy.

  3. Absorption and disposition of /sup 203/Hg in the pregnant and nonpregnant hamster following oral administration of (/sup 203/Hg)methylmercuric chloride

    SciTech Connect

    Reuhl, K.R.; Pounds, J.G.

    1981-02-01

    The absorption and disposition of /sup 203/Hg was determined in pregnant and nonpregnant golden hamsters by means of ..gamma.. scintillation spectroscopy. The animals were given 5 ..mu..Ci(/sup 203/Hg)methylmercuric chloride orally on gestation Day 9. Urine and feces were collected until Day 15, when the animals were sacrificed. Adult and fetal tissues were analyzed for /sup 203/Hg concentrations. At 12 h postintubation, pregnant hamsters absorbed more of the administered dose and eliminated less methylmercury in the feces than nonpregnant animals. Beyond 12 h postintubation, the absorption and elimination patterns were similar in both groups. Comparison of organs from pregnant and nonpregnant animals revealed greater methylmercury concentrations in the cerebella, pons-medullas, and spleens of pregnant females. The fetal head was found to contain significantly more methylmercury than the fetal body.

  4. Fabrication of {198Au0} radioactive composite nanodevices and their use for nano-brachytherapy

    PubMed Central

    Khan, Mohamed K.; Minc, Leah D.; Nigavekar, Shraddha S.; Kariapper, Muhammed S. T.; Nair, Bindu M.; Schipper, Matthew; Cook, Andrew C.; Lesniak, Wojciech G.; Balogh, Lajos P.

    2008-01-01

    We describe the simple fabrication of poly({198Au}) radioactive gold/dendrimer composite nanodevices in distinct sizes (between d=10 nm and 29 nm) for targeted radiopharmaceutical dose delivery to tumors in vivo. Irradiation of aqueous solutions of 197Au containing poly(amidoamine) dendrimer tetrachloroaurate salts or {197Au0} gold/dendrimer nanocomposites in a nuclear reactor resulted in the formation of positively charged and soluble poly{198Au0} radioactive composite nanodevices (CNDs). A mouse melanoma tumor model was used to test the poly{198Au0} CNDs whether they can deliver a therapeutic dose. A single intratumoral injection of poly{198Au0}d=22nm CNDs in PBS delivering a dose of 74 microCi, after eight days, resulted in a statistically significant 45% reduction in tumor volume, when compared to untreated groups and those injected with the “cold” nanodevice. No clinical toxicity was observed during the experiments. This study provides the first proof of principle that radioactive composite nanodevices can deliver therapeutic doses to tumors. PMID:18249156

  5. Determination of environmental dependence of the &-circ; decay half-life of ^198Au

    NASA Astrophysics Data System (ADS)

    Dibidad, A.; Goodwin, J.; Hardy, J.

    2009-10-01

    A series of articles by the C. Rolfs group [1] claimed changes in the half-lives of isotopes undergoing α, β^-, β^+, and electron-capture decays as the temperature reduced to 12 K from room temperature. These isotopes were contained in metallic, conductive environments, such as Au, Cu, and Pd, but it was also suggested that the half-life is different in an insulator. One publication [1] reported the half-life of ^198Au in a gold metal environment to change by 3.6 ±1.0% between room temperature and 12 K. Until then, radioactive half-lives were considered independent of environmental factors. We repeated the measurements of the ^198Au half-life in a gold metal environment under similar conditions as ref. [1] and demonstrated [2] that the half-life is the same at both temperatures within 0.04%, two orders of magnitude below the original claims. In the experiment reported here, we measured the half-life of ^198Au in an insulated environment -- gold (III) oxide -- at room temperature. Preliminary results indicate there is no difference in the measured half-life in an insulator as compared in a conductor. [4pt] [1] T. Spillane et al, Eur. Phys. J. A 31, 203 (2007) [0pt] [2] J.R. Goodwin et al, Eur. Phys. J. A 34, 271 (2007)

  6. Renal tubular handling of /sup 203/Hg/sup 2 +/ in the dog: a microinjection study

    SciTech Connect

    Cikrt, M.; Heller, J.

    1980-04-01

    /sup 203/HgCl/sub 2/ in trace amounts was injected together with (methoxy-/sup 3/H)inulin into the proximal convoluted tubules of the dog kidney superficial nephrons. Of the injected inulin, 97.3 +- 5.7% was recovered in the urine from the injected kidney. From the injected /sup 203/Hg only 8.2 +- 1.0% was recovered in the urine from the injected kidney and 0.4 +- 0.7% from the contralateral kidney. These results were not significantly influenced by intrarterial infusion of KCN, ouabain, or 2,4-dinitrophenol, indicating that there occurred no active tubular reabsorption of mercury. Of the injected /sup 203/Hg, 88.2 to 93.5% was recovered in the kidney whose tubules were punctured. When /sup 203/Hg was injected into peritubular capillary, only 1.2 +- 0.7% was recovered in the urine from the injected and 0.91 +- 0.2% from the contralateral kidney, this difference being not significant. Results are interpreted as lacking evidence for active transport mechanism of Hg/sup 2 +/ in the dog kidney under conditions of microinjection experiment.

  7. Meningosis prophylaxis with intrathecal /sup 198/Au-colloid and methotrexate in childhood acute lymphocytic leukemia

    SciTech Connect

    Metz, O.; Stoll, W.; Plenert, W.

    1982-01-15

    Since 1972, telecobalt irradiation plus intrathecal methotrexate (ITMTX) has been successfully replaced in Jena by intrathecal colloidal radioactive gold (/sup 198/Au) plus ITMTX for meningosis prophylaxis in leukemia. Seventy-three children with acute lymphocytic leukemia (ALL) were given 1.24-4.89 mCi (45.8-181 MBq) of colloidal 198Au IT after successful initiation of remission. During cytostatic therapy, the following relapses occurred: meningosis leucaemica, five patients (6.8%); bone-marrow relapse and the meningosis leucaemica, one patient; and bone-marrow relapse, 20 patients (27.4%). In 18 children, combination chemotherapy was terminated after two and a half or three years of treatment. After that time, one meningeal relapse and six bone-marrow relapses occurred. Within the first 24 hours after application of radioactive gold, headaches, vomiting, and fever occurred in less than 10% of the children. An apathy syndrome, leukecephalopathy, or severe infections, were not observed in a single case. Radioactive gold spreads in the subarachnoid space and is phagocytized by the arachnoidea. The tumoricide effect extends selectively over the space of distribution of the latent meningosis leucaemia. The cerebral parenchyma remains unaffected by radiation. Thus, radioactive gold may be preferable to telecobalt irradiation in preventing central nervous system leukemia.

  8. Absence of low-temperature dependence of the decay of {sup 7}Be and {sup 198}Au in metallic hosts

    SciTech Connect

    Kumar, V.; Hass, M.; Nir-El, Y.; Haquin, G.; Yungreiss, Z.

    2008-05-15

    The electron-capture (EC) decay rate of {sup 7}Be in metallic Cu host and the {beta}{sup -}-decay rate of {sup 198}Au in the host alloy Al-Au have been measured simultaneously at several temperatures, ranging from 0.350 K to 293 K. No difference of the half-life of {sup 198}Au between 12.5 K and 293 K is observed to a precision of 0.1%. By utilizing the special characteristics of our double-source assembly, possible geometrical effects that influence the individual rates could be eliminated. The ratio of {sup 7}Be to {sup 198}Au activity thus obtained also remains constant for this temperatures range to the experimental precision of 0.15{+-}0.16%. The resulting null temperature dependence is discussed in terms of the inadequacy of the often-used Debye-Hueckel model for such measurements.

  9. Absolute measurement of (198)Au activity in gold foil using plastic scintillators and a well-type NaI(Tl) detector.

    PubMed

    Kim, Yun Ho; Kim, Jungho; Lee, Jong-Man; Park, Hyeonseo

    2016-03-01

    A beta-gamma coincidence system has been developed for measuring (198)Au activity in gold foils. The system was validated by Monte Carlo simulations and by measuring the activity of a (60)Co point-source. To study effects such as self-shielding of beta particles in gold foils, (198)Au activity measurements and simulations were performed for various scintillators and foil sizes. The measured (198)Au activities were ~1% above the reference activity, which might be due to self-shielding of beta particles. The measured and simulated (198)Au activities agreed, suggesting feasibility of precise activity measurement.

  10. Laminin receptor specific therapeutic gold nanoparticles (198AuNP-EGCg) show efficacy in treating prostate cancer

    SciTech Connect

    Shukla, R.; Chanda, N.; Zambre, A.; Upendran, A.; Katti, K.; Kulkarni, R. R.; Nune, S. K.; Casteel, S. W.; Smith, C. J.; Vimal, J.; Boote, E.; Robertson, J. D.; Kan, P.; Engelbrecht, H.; Watkinson, L. D.; Carmack, T. L.; Lever, J. R.; Cutler, C. S.; Caldwell, C.; Kannan, R.; Katti, K. V.

    2012-07-16

    Systemic delivery of therapeutic agents to solid tumors is hindered by vascular and interstitial barriers. We hypothesized that prostate tumor specific epigallocatechingallate( EGCg) functionalized radioactive gold nanoparticles, when delivered intratumorally (IT), will circumvent transport barriers, resulting in targeted delivery of therapeutic payloads. The results described herein provide unequivocal validation of our hypothesis. We report the development of inherently therapeutic gold nanoparticles derived from Au-198 isotope; the range of 198Au β-particle ( ~ 11 mm in tissue or ~1100 cell diameters) is sufficiently long to provide cross-fire effects of radiation dose delivered to cells within the prostate gland and short enough to minimize radiation dose to critical tissues near the periphery of the capsule. The formulation of biocompatible 198AuNPs utilizes the redox chemistry of prostate tumor specific phytochemical EGCg as it converts gold salt into gold nanoparticles and also selectively binds with excellent affinity to Laminin67R receptors which are over expressed in prostate tumor cells. Pharmacokinetic studies in PC-3 xenograft SCID mice showed ~72% retention of 198AuNP-EGCg in tumors 24 h after intratumoral administration. Therapeutic studies showed 80% reduction of tumor volumes after 28 days demonstrating significant inhibition of tumor growth compared to controls. This innovative “green nanotechnological“approach serves as a basis for designing target specific antineoplastic agents. This novel intratumorally injectable 198AuNP-EGCg nanotherapeutic agent may provide significant advances in oncology for use as an effective treatment for prostate and other solid tumors.

  11. Laminin receptor specific therapeutic gold nanoparticles (198AuNP-EGCg) show efficacy in treating prostate cancer

    PubMed Central

    Shukla, Ravi; Chanda, Nripen; Zambre, Ajit; Upendran, Anandhi; Katti, Kavita; Kulkarni, Rajesh R.; Nune, Satish Kumar; Casteel, Stan W.; Smith, Charles Jeffrey; Vimal, Jatin; Boote, Evan; Robertson, J. David; Kan, Para; Engelbrecht, Hendrik; Watkinson, Lisa D.; Carmack, Terry L.; Lever, John R.; Cutler, Cathy S.; Caldwell, Charles; Kannan, Raghuraman; Katti, Kattesh V.

    2012-01-01

    Systemic delivery of therapeutic agents to solid tumors is hindered by vascular and interstitial barriers. We hypothesized that prostate tumor specific epigallocatechin-gallate (EGCg) functionalized radioactive gold nanoparticles, when delivered intratumorally (IT), would circumvent transport barriers, resulting in targeted delivery of therapeutic payloads. The results described herein support our hypothesis. We report the development of inherently therapeutic gold nanoparticles derived from the Au-198 isotope; the range of the 198Au β-particle (approximately 11 mm in tissue or approximately 1100 cell diameters) is sufficiently long to provide cross-fire effects of a radiation dose delivered to cells within the prostate gland and short enough to minimize the radiation dose to critical tissues near the periphery of the capsule. The formulation of biocompatible 198AuNPs utilizes the redox chemistry of prostate tumor specific phytochemical EGCg as it converts gold salt into gold nanoparticles and also selectively binds with excellent affinity to Laminin67R receptors, which are over expressed in prostate tumor cells. Pharmacokinetic studies in PC-3 xenograft SCID mice showed approximately 72% retention of 198AuNP-EGCg in tumors 24 h after intratumoral administration. Therapeutic studies showed 80% reduction of tumor volumes after 28 d demonstrating significant inhibition of tumor growth compared to controls. This innovative nanotechnological approach serves as a basis for designing biocompatible target specific antineoplastic agents. This novel intratumorally injectable 198AuNP-EGCg nanotherapeutic agent may provide significant advances in oncology for use as an effective treatment for prostate and other solid tumors. PMID:22802668

  12. Maternal-fetal distribution of mercury ( sup 203 Hg) released from dental amalgam fillings

    SciTech Connect

    Vimy, M.J.; Takahashi, Y.; Lorscheider, F.L. )

    1990-04-01

    In humans, the continuous release of Hg vapor from dental amalgam tooth restorations is markedly increased for prolonged periods after chewing. The present study establishes a time-course distribution for amalgam Hg in body tissues of adult and fetal sheep. Under general anesthesia, five pregnant ewes had twelve occlusal amalgam fillings containing radioactive 203Hg placed in teeth at 112 days gestation. Blood, amniotic fluid, feces, and urine specimens were collected at 1- to 3-day intervals for 16 days. From days 16-140 after amalgam placement (16-41 days for fetal lambs), tissue specimens were analyzed for radioactivity, and total Hg concentrations were calculated. Results demonstrate that Hg from dental amalgam will appear in maternal and fetal blood and amniotic fluid within 2 days after placement of amalgam tooth restorations. Excretion of some of this Hg will also commence within 2 days. All tissues examined displayed Hg accumulation. Highest concentrations of Hg from amalgam in the adult occurred in kidney and liver, whereas in the fetus the highest amalgam Hg concentrations appeared in liver and pituitary gland. The placenta progressively concentrated Hg as gestation advanced to term, and milk concentration of amalgam Hg postpartum provides a potential source of Hg exposure to the newborn. It is concluded that accumulation of amalgam Hg progresses in maternal and fetal tissues to a steady state with advancing gestation and is maintained. Dental amalgam usage as a tooth restorative material in pregnant women and children should be reconsidered.

  13. X-ray and gamma-ray intensity measurements in 131I, 166Ho, 198Au and 199Au decays

    NASA Astrophysics Data System (ADS)

    Chand, Bakhshish; Goswamy, Jatinder; Mehta, Devinder; Singh, Nirmal; Trehan, P. N.

    1989-12-01

    Relative intensities of X- and gamma rays emitted in the decays of 131I, 166Ho, 198Au and 199Au have been measured precisely using a set of five detectors: 96.0 cm 3 and 57.1 cm 3 coaxial HPGe detectors, a 28.27 mm 2 × 5.0 mm vertical planar HPGe detector and two 28.27 mm 2 × 5.5 mm Si(Li) detectors. The measured values of X-ray intensities are, in general, in good agreement with the theoretically calculated values. The intensities of different components of K and L X-rays in the above mentioned decays have been measured for the first time.

  14. Measurement of the half-life of {sup 198}Au in a nonmetal: High-precision measurement shows no host-material dependence

    SciTech Connect

    Goodwin, J. R.; Nica, N.; Iacob, V. E.; Dibidad, A.; Hardy, J. C.

    2010-10-15

    We have measured the half-life of the {beta}{sup -} decay of {sup 198}Au to be 2.6948(9) d, with the nuclide sited in an insulating environment. Comparing this result with the half-life we measured previously with a metallic environment, we find the half-lives in both environments to be the same within 0.04%, thus contradicting a prediction that screening from a ''plasma'' of quasifree electrons in a metal increases the half-life by as much as 7%.

  15. Gold nanoparticles production using reactor and cyclotron based methods in assessment of (196,198)Au production yields by (197)Au neutron absorption for therapeutic purposes.

    PubMed

    Khorshidi, Abdollah

    2016-11-01

    Medical nano-gold radioisotopes is produced regularly using high-flux nuclear reactors, and an accelerator-driven neutron activator can turn out higher yield of (197)Au(n,γ)(196,198)Au reactions. Here, nano-gold production via radiative/neutron capture was investigated using irradiated Tehran Research Reactor flux and also simulated proton beam of Karaj cyclotron in Iran. (197)Au nano-solution, including 20nm shaped spherical gold and water, was irradiated under Tehran reactor flux at 2.5E+13n/cm(2)/s for (196,198)Au activity and production yield estimations. Meanwhile, the yield was examined using 30MeV proton beam of Karaj cyclotron via simulated new neutron activator containing beryllium target, bismuth moderator around the target, and also PbF2 reflector enclosed the moderator region. Transmutation in (197)Au nano-solution samples were explored at 15 and 25cm distances from the target. The neutron flux behavior inside the water and bismuth moderators was investigated for nano-gold particles transmutation. The transport of fast neutrons inside bismuth material as heavy nuclei with a lesser lethargy can be contributed in enhanced nano-gold transmutation with long duration time than the water moderator in reactor-based method. Cyclotron-driven production of βeta-emitting radioisotopes for brachytherapy applications can complete the nano-gold production technology as a safer approach as compared to the reactor-based method.

  16. Metal transfer to plants grown on a dredged sediment: use of radioactive isotope 203Hg and titanium.

    PubMed

    Caille, Nathalie; Vauleon, Clotilde; Leyval, Corinne; Morel, Jean-Louis

    2005-04-01

    Improperly disposed of dredged sediments contaminated with metals may induce long-term leaching and an increase of metal concentrations in ground waters and vegetal cover plants. The objective of the study was to quantify the sediment-to-plant transfer of Cu, Pb, Hg and Zn with a particular focus on the pathway of Hg and to determine whether the establishment of vegetal cover modifies the metal availability. A pot experiment with rape (Brassica napus), cabbage (Brassica oleraccea) and red fescue (Festuca rubra) was set up using a sediment first spiked with the radioisotope 203Hg. Zinc concentrations (197-543 mg kg(-1) DM) in leaves were higher than Cu concentration (197-543 mg kg(-1) DM), Pb concentration (2.3-2.6 mg kg(-1) DM) and Hg concentration (0.9-1.7 mg kg(-1) DM). Leaves-to-sediment ratios decreased as follows: Zn > Cu > Hg > Pb. According to Ti measurements, metal contamination by dry deposition was less than 1%. Mercury concentration in plant leaves was higher than European and French thresholds. Foliar absorption of volatile Hg was a major pathway for Hg contamination with a root absorption of Hg higher in rape than in cabbage and red fescue. Growth of each species increased Cu solubility. Zinc solubility was increased only in the presence of rape. The highest increase of Cu solubility was observed for red fescue whereas this species largely decreased Zn solubility. Dissolved organic carbon (DOC) measurements suggested that Cu solubilisation could result from organic matter or release of natural plant exudates. Dissolved inorganic carbon (DIC) measures suggested that the high Zn solubility in the presence of rape could originate from a generation of acidity in rape rhizosphere and a subsequent dissolution of calcium carbonates. Consequently, emission of volatile Hg from contaminated dredged sediments and also the potential increase of metal solubility by a vegetal cover of grass when used in phytostabilisation must be taken into account by decision

  17. Isomeric ratios for ^{196,198}Tl and ^{196,198}Au from fusion and transfer in the interaction of 6He with 197Au

    NASA Astrophysics Data System (ADS)

    Kulko, A. A.; Demekhina, N. A.; Kalpakchieva, R.; Kolesnikov, N. N.; Lukashik, V. G.; Penionzhkevich, Yu E.; Rassadov, D. N.; Skobelev, N. K.

    2007-11-01

    Excitation functions and isomeric cross-section ratios have been measured for the isomeric pairs 198m,gTl, 196m,gTl, 198m,gAu and 196m,gAu, formed in fusion-evaporation and transfer processes in reactions of 6He with 197Au in the energy range 7-60 MeV. The population probability of high- and low-spin states in the fusion reaction with the formation of 198Tl and 196Tl (Jπm = 7+, Jπg = 2-), as well as of the transfer products 198Au and 196Au (Jπm = 12-, Jπg = 2-) is compared with calculations within the statistical model and with similar results, obtained in reactions induced by other nuclei.

  18. Validation of absolute axial neutron flux distribution calculations with MCNP with 197Au(n,γ)198Au reaction rate distribution measurements at the JSI TRIGA Mark II reactor.

    PubMed

    Radulović, Vladimir; Štancar, Žiga; Snoj, Luka; Trkov, Andrej

    2014-02-01

    The calculation of axial neutron flux distributions with the MCNP code at the JSI TRIGA Mark II reactor has been validated with experimental measurements of the (197)Au(n,γ)(198)Au reaction rate. The calculated absolute reaction rate values, scaled according to the reactor power and corrected for the flux redistribution effect, are in good agreement with the experimental results. The effect of different cross-section libraries on the calculations has been investigated and shown to be minor.

  19. Thermal neutron calibration of a tritium extraction facility using the /sup 6/Li(n,t)/sup 4/He//sup 197/Au(n,. gamma. )/sup 198/Au cross section ratio for standardization

    SciTech Connect

    Bretscher, M.M.; Smith, D.L.

    1980-08-01

    Absolute tritium activities in a neutron-activated metallic lithium samples have been measured by liquid scintillation methods to provide data needed for the determination of capture-to-fission ratios in fast breeder reactor spectra and for recent measurements of the /sup 7/Li(n,n't)/sup 4/He cross section. The tritium extraction facility used for all these experiments has now been calibrated by measuring the /sup 6/Li(n,t)/sup 4/He//sup 197/Au/n,..gamma..)/sup 198/Au activity ratio for thermal neutrons and comparing the result with the well-known cross sections. The calculated-to-measured activity ratio was found to be 1.033 +- 0.018. 2 figures, 20 tables.

  20. Treatment of malignant pericardial effusion with 32P-colloid

    PubMed Central

    Dempke, W; Firusian, N

    1999-01-01

    Malignant pericardial effusion is usually treated only when signs of cardiac tamponade develop. Several methods of treatment have been reported with an overall response rate of approximately 75%. Since our initial study using intrapericardial 32P-colloid instillation as a treatment modality for pericardial effusion demonstrated a significant higher response rate, this study was conducted to further evaluate the efficacy of intrapericardial 32P-colloid in terms of response rates and duration of remissions. Intrapericardial instillation of 185–370 MBq (5–10 mCi) 32P-colloid in 36 patients with malignant pericardial effusion resulted in a complete remission rate of 94.5% (34 patients) whereas two patients did not respond to treatment due to a foudroyant formation of pericardial fluid. The median duration time was 8 months. No side-effects were observed. These results suggest that intrapericardial instillation of 32P-colloid is a simple, reliable and safe treatment strategy for patients with malignant pericardial effusions. Therefore, since further evidence is provided that 32P-colloid is significantly more effective than external radiation or non-radioactive sclerosing agents, this treatment modality should be considered for the management of malignant pericardial effusion. © 1999 Cancer Research Campaign PMID:10471044

  1. [Diagnosis of uveitis with fractional doses of 32-P radionuclide].

    PubMed

    Shambra, V V; Panfilova, G V

    1989-01-01

    For determination of the presence and activity of inflammation in the vascular coat of the eye as well as of the stopping of the disease, the authors propose to use radionuclide 32-P in a form of fractional doses in contradistinction to a single administration of a conventional dose. A common dose of the preparation is divided into four equal parts. At first, 1/4 of the dose is administered. One hour later, comparative external radiometry of the patient and the healthy eye is made. If there is seen no clear difference in intensity of accumulation, the second dose is administered. One hour later, repeated examination is made. After receiving clear data, further administration of the preparation is stopped. Administration of fractional doses suggests a decrease of ray load on the patient's organism, economy of the preparation without lowering the authenticity of examination.

  2. Suppression of hepatic hematopoiesis with radioactive gold (/sup 198/Au)

    SciTech Connect

    Turner, A.R.; Gummerman, L.W.; Boggs, D.R.

    1985-04-01

    A patient with idiopathic myelofibrosis of some 20 yr duration developed esophageal varices and ascites. No explanation for increased portal pressure other than hepatic hematopoiesis was found. Consequently, a trial of cobalt irradiation to the liver was undertaken with definite but transient decrease in ascites. Subsequently, two courses of radioactive colloidal gold were given, again with definite but transient beneficial effects on the degree of ascites. This latter benefit occurred without suppression of marrow function.

  3. An overview of DNA fingerprinting with sup 32 P nucleotides

    SciTech Connect

    Pappas, G.G.

    1992-01-01

    The DNA probes radiolabeled with {sup 32}P, a primary tool employed by researchers in the life sciences for > 20 yr, are used by private companies, state-run laboratories, and the FBI to generate autoradiographs displaying the unique banding patterns that constitute the DNA fingerprint. The ability to identify an individual or animal from a biological sample has profound implications. Unidentified bodies, unrecognizable remains, and missing children can be tested and the DNA fingerprint compared to those of family members for positive identification. Paternity can be established before a child's birth. Immigration disputes can easily be resolved. Other uses include pedigree determination and testing for cell-line cross-contamination. Using a DNA fingerprint to determine the guilt or innocence of an individual allegedly involved in a violent crime is very controversial and has great legal and moral implications for society. Forensic laboratories have been challenged to ensure a level of quality control and quality assurance consistent with the weight given to these tests when used as evidence in a court of law.

  4. Determination of the 32P activity in angioplastic balloons by LSC.

    PubMed

    Günther, E

    2002-01-01

    The determination of the 32P activity in angioplastic balloons involves two problems: the extraction of 32P, which is covered with plastic foils, and the determination of the 33P impurity. At PTB the active balloons are destroyed by combustion in an oxygen stream. The active phosphorus is extracted quantitatively from the tube. The activities of 32P and 33P are determined by measurements performed over a period of one month or more by a subsequent data fit.

  5. Radiophosphorus (/sup 32/P) treatment of bone marrow disorders in dogs: 11 cases (1970-1987)

    SciTech Connect

    Smith, M.; Turrel, J.M.

    1989-01-01

    Between March 1970 and February 1987, radiophosphorus (/sup 32/P) was used to treat bone marrow disorders in 6 dogs; 4 had polycythemia vera and 2 had essential thrombocythemia. Activities of /sup 32/P given initially ranged from 2.4 to 3.3 mCi/m2. Four dogs responded well to /sup 32/P treatment, with gradual resolution of high RBC or platelet counts. Two of these dogs died of intercurrent disease unrelated to their bone marrow disorder, before blood counts could be stabilized. Two dogs did not respond to the initial /sup 32/P treatment nor to additional treatments with /sup 32/P, and had clinical signs and blood counts stabilized by use of phlebotomy or chemotherapeutic agents. We reviewed and analyzed 5 other cases of bone marrow disorders in dogs treated with /sup 32/P and included the findings from their records with the records of our 6 dogs in this retrospective analysis. Of the 8 dogs with polycythemia vera treated with /sup 32/P, 5 were given a single treatment that controlled clinical signs and blood counts for the remainder of the follow-up period. Of the 3 dogs treated for thrombocytosis with /sup 32/P, 2 had blood counts that responded to a single treatment.

  6. A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate.

    PubMed

    Parra, F

    1982-09-01

    A rapid and simple enzymic method is described for the synthesis of [(32)P]phosphoenolpyruvate from [(32)P]P(i), with a reproducible yield of 74%. The final product was shown to be a good substrate for pyruvate kinase (EC 2.7.1.40).

  7. A simple enzymic method for the synthesis of [32P]phosphoenolpyruvate

    PubMed Central

    Parra, Francisco

    1982-01-01

    A rapid and simple enzymic method is described for the synthesis of [32P]phosphoenolpyruvate from [32P]Pi, with a reproducible yield of 74%. The final product was shown to be a good substrate for pyruvate kinase (EC 2.7.1.40). ImagesFig. 1. PMID:7150238

  8. Determination of phagocytosis of /sup 32/P-labeled Staphylococcus aureus by bovine polymorphonuclear leukocytes

    SciTech Connect

    Dulin, A.M.; Paape, M.J.; Weinland, B.T.

    1984-04-01

    A procedure for the measurement of phagocytosis by bovine polymorphonuclear leukocytes (PMN) of /sup 32/P-labeled Staphylococcus aureus was modified so that a larger number of samples could be compared in a single run, and smaller volumes of sample, PMN, and /sup 32/P-labeled S aureus could be used. Results were highly reproducible, with a coefficient of variation between duplicate determinations of less than or equal to 2%. Lysostaphin was prepared from the supernatant of S staphylolyticus and was compared with a commercially available preparation. Effects of lysostaphin on PMN and influence of incubation media on release of /sup 32/P from /sup 32/P-labeled S aureus by lysostaphin were examined.

  9. Recalculation of data on 32P activity induced in sulfur in Hiroshima.

    PubMed

    Hamada, T

    1991-03-01

    Historical data for 32P activity induced in sulfur by fast neutrons have been corrected for decay with a recent half-life value of 32P and recalculated with an experimentally determined efficiency ratio of the electroscope for beta rays from 32P and natural uranium used as a standard. Most samples would have been pure enough so that no correction for the weight of sulfur has been made. The possibility of interference with 32P activity measurements due to induced activity of other elements in the samples could also be excluded. The revised data show little difference from the original ones except for one sample which contained much impurity. Uncertainty of the data was also discussed.

  10. 32P-postlabeling analysis of non-radioactive aromatic carcinogen--DNA adducts.

    PubMed

    Gupta, R C; Reddy, M V; Randerath, K

    1982-01-01

    A newly developed enzymatic 32P-postlabeling method was applied to the analysis of DNA's containing non-radioactive arylamine, arylamide, and polycyclic aromatic hydrocarbon adducts. DNA reacted in vitro with N-hydroxy-2-amino-fluorene, N-acetoxy-2-acetylaminofluorene, and 7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, respectively, as well as DNA preparations from the liver of rats treated with N-hydroxy-2-acetylaminofluorene and benzo[a]pyrene, respectively, were enzymatically digested to deoxyribonucleoside 3'-monophosphates, which were then converted to [5'-32P]deoxyribonucleoside 3',5'-bisphosphates by T4 polynucleotide kinase-catalyzed [32P]phosphate transfer from [gamma-32P]ATP. The 32P-labeled nucleotides were resolved by anion-exchange t.l.c. on polyethyleneimine-cellulose and detected by autoradiography. Aromatic adduct nucleotides were found to be retained at the origin in aqueous electrolyte solutions, but to migrate as distinct spots in solvents containing 7-8.5 M urea. Advantage was taken of this observation to remove 32P-labeled normal DNA nucleotides from adduct nucleotides. This purification enabled the detection of a single adduct in 10(7)-10(8) normal nucleotides. The method appears applicable to the ultrasensitive detection of a large number of carcinogen--DNA adducts of diverse structure without requiring radioactive carcinogens or specific antibodies.

  11. The analysis of DNA adducts: The transition from 32P-postlabeling to mass spectrometry

    PubMed Central

    Klaene, Joshua J.; Sharma, Vaneet K.; Glick, James; Vouros, Paul

    2012-01-01

    The technique of 32P-postlabeling, which was introduced in 1982 for the analysis of DNA adducts, has long been the method of choice for in vivo studies because of its high sensitivity as it requires only <10 μg DNA to achieve the detection of 1 adduct in 1010 normal bases. 32P-postlabeling has therefore been utilized in numerous human and animal studies of DNA adduct formation. Like all techniques 32P-postlabeling does have several disadvantages including the use of radioactive phosphorus, lack of internal standards, and perhaps most significantly does not provide any structural information for positive identification of unknown adducts, a shortcoming that could significantly hamper progress in the field. Structural methods have since been developed to allow for positive identification of DNA adducts, but to this day, the same level of sensitivity and low sample requirements provided by 32P-postlabeling have not been matched. In this mini review we will discuss the 32P-postlabeling method and chronicle the transition to mass spectrometry via the hyphenation of gas chromatography, capillary electrophoresis, and ultimately liquid chromatography which, some 30 years later, is only just starting to approach the sensitivity and low sample requirements of 32P-postlabeling. This paper focuses on the detection of bulky carcinogen-DNA adducts, with no mention of oxidative damage or small alkylating agents. This is because the 32P-postlabeling assay is most compatible with bulky DNA adducts. This will also allow a more comprehensive focus on a subject that has been our particular interest since 1990. PMID:22960573

  12. Laboratory and field studies with /sup 32/P labeled Toxorhynchites rutilus rutilus

    SciTech Connect

    Smittle, B.J.; Focks, D.A.

    1986-12-01

    Females and eggs of Toxorhynchites r. rutilus were labeled with /sup 32/P by feeding fourth-stage larvae /sup 32/P labeled Aedes aegypti larvae. Eggs from females up to 3 weeks in age had detectable levels of radioactivity and individual eggs contained ca. 0.3% of the mother's total radioactivity. Comparisons of labeled and unlabeled females in indoor and outdoor cage tests indicated that survival and fecundity of the 2 groups were approximately equal. No differences were noted for dispersal and fecundity of labeled and control females released in field tests. The /sup 32/P-labeled Tx. r. rutilus females behave similarly to unlabeled females, and this method of radiolabeling provides a sound tool for tracking laboratory-reared females released into an area with an indigenous population.

  13. Absorption and elimination kinetics of 32P malathion in the hen.

    PubMed

    Gupta, P K; Paul, B S; Ehrnebo, M

    1977-01-01

    A method for the separation of urine and faeces in the hen was developed and the biological fate of 32P malathion, following a single oral dose of 262.40 mg/kg body weight was studied. The results suggested that the compound was rapidly absorbed from the gastrointestinal tract; significant quantities being detected in the plasma and whole blood 1/2 hr after ingestion. Total 32P was eliminated in the urine by apparent first order kinetics with an average half-life of 5.7 hrs. The cumulative urinary and faecal data revealed that 93% of the 32P is excreted via the urine within 48 hrs, thus indicating that the compound is almost completely absorbed. It is therefore concluded that accumulation in the system is unlikely.

  14. Validation and verification of the ICRP biokinetic model of 32P: the criticality accident at Tokai-Mura, Japan.

    PubMed

    Miyamoto, K; Takeda, H; Nishimura, Y; Yukawa, M; Watanabe, Y; Ishigure, N; Kouno, F; Kuroda, N; Akashi, M

    2003-01-01

    Regrettably, a criticality accident occurred at a uranium conversion facility in Tokai-mura, Ibaraki, Japan, on 30 September 1999. Radioactivities of 32P in urine, blood and bone samples of the victims, who were severely exposed to neutrons, were measured. 32P was induced in their whole bodies at the moment of the first nuclear release by the reaction 31P (n, gamma) 32P and 32S (n, p) 32P. A realistic biokinetic model was assumed, as the exchange of 32P between the extracellular fluid compartment and the soft tissue compartment occurs only through the intracellular compartment, and the model was used for preliminary calculations. Some acute excretion of 32P, caused by decomposition or elution of tissues which occurred at the time of the accident, may have happened in the victims' bodies in the first few days. The working hypotheses in the present work should initiate renewed discussion of 32P biokinetics.

  15. A method for the 32P labeling of peptides or peptide nucleic acid oligomers

    NASA Technical Reports Server (NTRS)

    Kozlov, I. A.; Nielsen, P. E.; Orgel, L. E.; Bada, J. L. (Principal Investigator)

    1998-01-01

    A novel approach to the radioactive labeling of peptides and PNA oligomers is described. It is based on the conjugation of a deoxynucleoside 3'-phosphate with the terminal amine of the substrate, followed by phosphorylation of the 5'-hydroxyl group of the nucleotide using T4 polynucleotide kinase and [gamma-32P]ATP.

  16. A simple enzymic method for the synthesis of adenosine 5'-[alpha-32P]triphosphate on a preparative scale.

    PubMed

    Martin, B R; Voorheis, H P

    1977-03-01

    A simple, rapid and inexpensive method is described for the enzymic synthesis of [alpha-32P]ATP from [32P]Pi on a preparative scale with an overall yield of 53%. The final product contained all of the detectable radioactivity (less than 99.9%) in the alpha position and has been shown to behave identically with commerically availabe [alpha-32P]ATP during the synthesis of 3':5'-cyclic AMP in the reaction catalysed by adenylate cyclase.

  17. Estimates of intakes and internal doses from ingestion of {sup 32}P at MIT and NIH

    SciTech Connect

    Stabin, M.G.; Toohey, R.E.

    1996-06-01

    A researcher at Massachusetts Institute of Technology (MIT) became internally contaminated with {sup 32}P, probably due to an intentional act. The incident occurred on or about 14 August 1995. Subsequent measurement of activity in urine and a single whole body count were used to estimate the individual`s intake, with the assumption of ingestion as the route of intake. Two separate Sets of urine data were analyzed-one supplied by MIT and one from independent analyses of urine samples conducted at Oak Ridge Institute for Science and Education (ORISE); the former data set contained 35 samples, the latter 49. In addition, the results of 35 whole body counts, provided by MIT from a chair-type counter calibrated for 32p, were used to obtain a separate estimate of intake. The kinetic model for 32P proposed in ICRP Publication 30 and implemented in NUREG/CR-4884 was used to interpret the data. The data were analyzed using both the weighted and unweighted least squares techniques. All of the intake estimates were in very good agreement with each other, ranging from 18-22 MBq. Based on the dose model in ICRP 30, this would indicate a committed effective dose equivalent of 38-46 mSv. The incident was helpful in assessing the value of the least squares techniques in determining estimates of intake and dose. The ICRP model tended to slightly overestimate the whole body retention data and underestimate the urinary excretion at later times. Further results obtained by visual best fit and development of an individual-specific kinetic and dose model will also be discussed. This incident was quite similar to another case of ingestion of 32p that occurred at the National Institute of Health (NIH) on 28 June 1995. Dose assessment for the NIH case will also be presented if the data are available for public release.

  18. Colloidal chromic phosphate /sup 32/P synovectomy in antigen-induced arthritis in the rabbit

    SciTech Connect

    Howson, M.P.; Shepard, N.L.; Mitchell, N.S.

    1988-04-01

    Radioisotopes have been employed in the therapy of chronic arthritis, in particular, rheumatoid arthritis for many years. A variety of isotopes have been popularized, and in the last ten years a colloidal solution of radioactive chromic phosphate /sup 32/P has been in use apparently with equivalent efficacy to others such as /sup 169/erbium, /sup 90/yttrium, and /sup 165/dysprosium. No controlled studies on this modality have been reported and few animal studies were found. The efficacy of therapeutic doses of /sup 32/P as a medical synovectomy and its effect on rabbit joints with antigen-induced arthritis were observed in 62 arthritic knee joints in 31 adult rabbits treated on one side with 0.1 microCi of /sup 32/P, the opposite serving as control. The animals were observed over a period of 11 months and examined by histologic and biochemical means. The synovium showed no evidence of radiation necrosis in treated joints. Cartilage of treated and control joints showed similar changes consistent with chronic arthritis, persistent synovitis, progressive chondrocyte degeneration, and decreased matrix metachromasia. The radiosynovectomy had neither removed synovium nor protected the cartilage. Its efficacy in humans is therefore questionable.

  19. Dose-rate distribution of {sup 32}P-glass microspheres for intra-arterial brachytherapy

    SciTech Connect

    Guimaraes, Carla C.; Moralles, Mauricio; Sene, Frank F.; Martinelli, Jose R.

    2010-02-15

    Purpose: The intra-arterial administration of radioactive glass microspheres is an alternative therapy option for treating primary hepatocellular carcinoma, the main cause of liver cancer death, and metastatic liver cancer, another important kind of cancer induced in the liver. The technique involves the administration of radioactive microspheres in the hepatic artery, which are trapped preferentially in the tumor. Methods: In this work the GEANT4 toolkit was used to calculate the radial dose-rate distributions in water from {sup 32}P-loaded glass microspheres and also from {sup 90}Y-loaded glass microspheres. To validate the toolkit for this application, the authors compared the dose-rate distribution of {sup 32}P and {sup 90}Y point sources in water with data from the International Commission on Radiation Units and Measurements report 72. Results: Tables of radial dose-rate distributions are provided for practical use in brachytherapy planning with these microspheres. Conclusions: The simulations with the microspheres show that the shape of the beta ray energy spectra with respect to the {sup 32}P and {sup 90}Y sources is significantly modified by the glass matrix.

  20. Preparation of (32)P-end-labeled DNA fragments for performing DNA-binding experiments.

    PubMed

    Carey, Michael F; Peterson, Craig L; Smale, Stephen T

    2013-05-01

    The generation of a uniquely (32)P-end-labeled DNA fragment is essential for DNA-binding experiments such as DNase I footprinting and ethylation interference. We describe here a protocol for end-labeling a restriction fragment. For a plasmid DNA bearing a region containing the binding site of interest, cleaving with a single restriction endonuclease generates a 5' overhang containing a phosphate. This is generally necessary for both common forms of fragment end-labeling: phosphorylation with polynucleotide kinase and "filling in the end" with DNA polymerases (e.g., Klenow fragment). For the phosphorylation reaction, as described here, the phosphate is removed with calf intestinal phosphatase or bacterial alkaline phosphatase, and the resulting free 5'-OH is phosphorylated with polynucleotide kinase and [γ-(32)P]ATP. This generates a plasmid labeled at each end with γ-(32)P. The molar amount of plasmid DNA must be below the amount of ATP added to the reaction and the ATP must be of sufficiently high specific activity to generate a fragment labeled to the extent necessary for many DNA-binding experiments. To generate a uniquely end-labeled DNA fragment, the labeled plasmid is heat-treated to inactivate any remaining kinase and recleaved with a second endonuclease, releasing a short DNA fragment and a longer vector fragment. The DNA fragment is purified from the labeled vector on a 5%-8% native polyacrylamide gel. The preparation and labeling of DNA restriction fragments typically takes 1-2 d.

  1. Radial 32P ion implantation using a coaxial plasma reactor: Activity imaging and numerical integration

    NASA Astrophysics Data System (ADS)

    Fortin, M. A.; Dufresne, V.; Paynter, R.; Sarkissian, A.; Stansfield, B.

    2004-12-01

    Beta-emitting biomedical implants are currently employed in angioplasty, in the treatment of certain types of cancers, and in the embolization of aneurysms with platinum coils. Radioisotopes such as 32P can be implanted using plasma-based ion implantation (PBII). In this article, we describe a reactor that was developed to implant radioisotopes into cylindrical metallic objects. The plasma first ionizes radioisotopes sputtered from a target, and then acts as the source of particles to be implanted into the biased biomedical device. The plasma therefore plays a major role in the ionization/implantation process. Following a sequence of implantation tests, the liners protecting the interior walls of the reactor were changed and the radioactivity on them measured. This study demonstrates that the radioactive deposits on these protective liners, adequately imaged by radiography, can indicate the distribution of the radioisotopes that are not implanted. The resulting maps give unique information about the activity distribution, which is influenced by the sputtering of the 32P-containing fragments, their ionization in the plasma, and also by the subsequent ion transport mechanisms. Such information can be interpreted and used to significantly improve the efficiency of the implantation procedure. Using a surface barrier detector, a comparative study established a relationship between the gray scale of radiographs of the liners, and activity measurements. An integration process allows the quantification of the activities on the walls and components of the reactor. Finally, the resulting integral of the 32P activity is correlated to the sum of the radioactivity amounts that were sputtered from radioactive targets inside the implanter before the dismantling procedure. This balance addresses the issue of security regarding PBII technology and confirms the confinement of the radioactivity inside the chamber.

  2. Radial {sup 32}P ion implantation using a coaxial plasma reactor: Activity imaging and numerical integration

    SciTech Connect

    Fortin, M.A.; Dufresne, V.; Paynter, R.; Sarkissian, A.; Stansfield, B.

    2004-12-01

    Beta-emitting biomedical implants are currently employed in angioplasty, in the treatment of certain types of cancers, and in the embolization of aneurysms with platinum coils. Radioisotopes such as {sup 32}P can be implanted using plasma-based ion implantation (PBII). In this article, we describe a reactor that was developed to implant radioisotopes into cylindrical metallic objects. The plasma first ionizes radioisotopes sputtered from a target, and then acts as the source of particles to be implanted into the biased biomedical device. The plasma therefore plays a major role in the ionization/implantation process. Following a sequence of implantation tests, the liners protecting the interior walls of the reactor were changed and the radioactivity on them measured. This study demonstrates that the radioactive deposits on these protective liners, adequately imaged by radiography, can indicate the distribution of the radioisotopes that are not implanted. The resulting maps give unique information about the activity distribution, which is influenced by the sputtering of the {sup 32}P-containing fragments, their ionization in the plasma, and also by the subsequent ion transport mechanisms. Such information can be interpreted and used to significantly improve the efficiency of the implantation procedure. Using a surface barrier detector, a comparative study established a relationship between the gray scale of radiographs of the liners, and activity measurements. An integration process allows the quantification of the activities on the walls and components of the reactor. Finally, the resulting integral of the {sup 32}P activity is correlated to the sum of the radioactivity amounts that were sputtered from radioactive targets inside the implanter before the dismantling procedure. This balance addresses the issue of security regarding PBII technology and confirms the confinement of the radioactivity inside the chamber.

  3. Effect of lithosperm on thyroidal /sup 32/P uptake at various times of injection

    SciTech Connect

    Breneman, W.R.; Zeller, F.J.

    1983-06-01

    These experiments were performed to increase our understanding of possible side effects in the use of extracts of the plant Lithospermum ruderale (LSPM) as a contraceptive. Cold-water extracts of LSPM were used to note possible effects on injected TSH and on endogenous TSH which was increased by the use of propylthiouracil. It was demonstrated that LSPM had a biphasic effect on both endogenous and exogenous TSH activity as measured by chick thyroid /sup 32/P uptake. When given 18h before autopsy, LSPM decreased TSH activity in both, whereas when LSPM was administered 42h or 44h before autopsy, TSH activity was significantly increased.

  4. Formation of DNA adducts from oil-derived products analyzed by 32P-HPLC.

    PubMed

    Akkineni, L K; Zeisig, M; Baranczewski, P; Ekström, L G; Möller, L

    2001-01-01

    The aim of this study was to investigate the genotoxic potential of DNA adducts and to compare DNA adduct levels and patterns in petroleum vacuum distillates, coal tar distillate, bitumen fume condensates, and related substances that have a wide range of boiling temperatures. An in vitro assay was used for DNA adduct analysis with human and rat S-9 liver extract metabolic activation followed by 32P-postlabeling and 32P-high-performance liquid chromatography (32p-HPLC). For petroleum distillates originating from one crude oil there was a correlation between in vitro DNA adduct formation and mutagenic index, which showed an increase with a distillation temperature of 250 degrees C and a peak around a distillation point of approximately 400 degrees C. At higher temperatures, the genotoxicity (DNA adducts and mutagenicity) rapidly declined to very low levels. Different petroleum products showed a more than 100-fold range in DNA adduct formation, with severely hydrotreated base oil and bitumen fume condensates being lowest. Coal tar distillates showed ten times higher levels of DNA adduct formation than the most potent petroleum distillate. A clustered DNA adduct pattern was seen over a wide distillation range after metabolic activation with liver extracts of rat or human origin. These clusters were eluted in a region where alkylated aromatic hydrocarbons could be expected. The DNA adduct patterns were similar for base oil and bitumen fume condensates, whereas coal tar distillates had a wider retention time range of the DNA adducts formed. Reference substances were tested in the same in vitro assay. Two- and three-ringed nonalkylated aromatics were rather low in genotoxicity, but some of the three- to four-ringed alkylated aromatics were very potent inducers of DNA adducts. Compounds with an amino functional group showed a 270-fold higher level of DNA adduct formation than the same structures with a nitro functional group. The most potent DNA adduct inducers of the 16

  5. 32P-postlabelling of diastereomeric 7-alkylguanine adducts of butadiene monoepoxide.

    PubMed

    Kumar, R; Vodicka, P; Koivisto, P; Peltonen, K; Hemminki, K

    1996-06-01

    The reaction of 3,4-epoxy-1-butene (BMO) with deoxyguanosine-3'-monophosphate (3'-dGMP) resulted in the formation of two pairs of diastereomeric 7-alkyl-3'-dGMP derivatives corresponding to two isomers C¿-1 and C¿-2. The T4 polynucleotide kinase-mediated phosphorylation with [gamma-32P]-ATP showed preferential labelling of diastereo- mers of the C¿-1 isomer. The diastereomers 1 and 2 of the C¿-1 isomer had labelling efficiencies of 42%. However, the labelling efficiencies of diastereomers 3 and 4 of the C¿-2 isomer were 11 and 10%, respectively. The 32P-postlabelling of BMO-modified DNA yielded four isomers in the ratio of 4:4:1:1 with overall recoveries being 14%. The two isomers had a half-life of 270 min (C¿-1 isomer) and 300 min (C¿-2 isomer) which is in accordance with the stability predicted by other similar adduct experiments. The molecular modelling experiments showed more pronounced restricted rotation of butadiene residue in C¿-2 isomers due to steric interaction between butadiene residue at N-7 and O(6) atom of guanine than in C¿-1 isomer. The butadiene residue also leads to steric overcrowding at 3'-phosphate in C¿-2 isomer which probably restricts the access to the active site of T4 polynucleotide kinase.

  6. Evaluation of Isotope 32P Method to Mark Culex pipiens (Diptera: Culicidae) in a Laboratory

    PubMed Central

    Zhang, Chongxing; Shi, Guihong; Zhao, Yuqiang; Yan, Dongmei; Li, Huaiju; Liu, Hongmei; Wiwatanaratanabutr, Itsanun; Gong, Maoqing

    2016-01-01

    Background: The aim of the current study was to develop a marking technique as an internal marker to mark post blood meal mosquitoes by using stable phosphate isotope 32P and determine the optimal concentration of it. Methods: An isotonic physiological saline solution, containing different concentration of radioactive isotope 32P-labeled disodium phosphate (Na2H32PO4) was injected into rabbits via the jugular vein in the laboratory. Emerged Cx. pipiens were marked after feeding on rabbit. At the same time, the labeled conditions of emerged Cx. pipiens were also measured by placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside. Results: According to the label condition of Cx. pipiens after taking blood and the effect of different dosage Na2H32PO4 on rabbit health, the optimal concentration of radioactive isotope was determined, that is, 0.1211 mCi/kg. By placing feces of No. 6 rabbit into containers with mosquito larvae and pupae inside, the emerged mosquitoes were also labeled. Therefore, feeding mosquitoes on the animal injected with radioactive Na2H32PO4 was more practical for detecting and tracing mosquitoes. Conclusion: The method was less time-consuming, more sensitive and safer. This marking method will facilitate post-bloodmeal studies of mosquitoes and other blood-sucking insects. PMID:27308279

  7. (32)P colloid radiosynovectomy in treatment of chronic haemophilic synovitis: Iran experience.

    PubMed

    Mortazavi, S M J; Asadollahi, S; Farzan, M; Shahriaran, S; Aghili, M; Izadyar, S; Lak, M

    2007-03-01

    Repeated intra-articular bleeding with subsequent development of chronic synovitis and cartilage changes, leading to haemophilic arthropathy, is one the most debilitating problems in haemophilic patients. Radiosynovectomy is a familiar therapeutic choice in management of chronic synovitis in haemophilia. We report the treatments results of synoviorthesis with (32)P chromic phosphate with emphasis on clinical aspects. Between 2002 and 2006 we performed 66 procedures in 53 patients. Seven patients were excluded. The remaining 46 patients were followed for an average of 31 months. The mean age of patients at the time of injection was 15.9 years (range: 6-28). There were three repeat injections. According to Fernandez-pallazi and Cavilgia clinical classification (Table 1) [23], nine joints were Stage II and 46 were Stage III. In latest follow-up, 77% of patients reported at least a 50% decrease in bleeding frequency after treatment (P < 0.0001). The need for antihaemophilic factor consumption dropped by about 74% postradiosynovectomy (P < 0.0001). In most of the injected joints, the range of motion remained stable or improved. A trend was found for the number of haemarthrosis to increase after a period of considerable improvement. Synoviorthesis using (32)P effectively reduces the intra-articular bleeding rate and factor concentrate use. Durability of the response seems to be unpredictable, perhaps attributable to the late intervention. An early radiosynovectomy might be more helpful in terms of stability of response to treatment.

  8. Incorporation of 32P and 14C into Photosynthetic Products of Ankistrodesmus braunii as Affected by X-Rays

    PubMed Central

    Jeschke, W. D.; Gimmler, H.; Simonis, W.

    1967-01-01

    The incorporation of 32P and 14C into organic compounds by Ankistrodesmus is strongly inhibited by X-rays. In the same phosphorylated compounds 32P-incorporation apparently is more severely inhibited by X-rays than the 14C-labelling. The 32P-incorporation into organic compounds is more strongly inhibited than 32P-labelling of inorganic phosphate in the cell. The inhibition of 32P-incorporation into a number of compounds is strikingly uniform. It is concluded that the inhibition of 32P-incorporation and of 14C-incorporation into phosphorylated compounds in vivo is due to an uncoupling by X-rays of photophosphorylation as in vitro. The difference in X-ray sensitivity of 14C- and 32P-incorporation into one organic phosphorous compound is attributed to a dual action of X-rays on 32P-incorporation in organic compounds (both via the uncoupling of photophosphorylation) and only a single effect on 14C-incorporation and 32P-labelling of inorganic phosphate. The effect of X-rays on 14C-incorporation into organic compounds included inhibition in most cases but also stimulation as in the case of glycolic acid. These differences may be due to interference in the intercellular regulations following the application of X-rays. The inhibition of 14C-incorporation in many cases exhibits different behaviour at low (<200 krad) and high doses. These changes are discussed on the assumption that at the lower doses X-rays cause uncoupling of photophosphorylation and at the higher doses an additional inhibition of electron transport. PMID:16656516

  9. Pediatric dosimetry for intrapleural lung injections of (32)P chromic phosphate.

    PubMed

    Konijnenberg, Mark W; Olch, Arthur

    2010-10-07

    Intracavitary injections of (32)P chromic phosphate are used in the therapy of pleuropulmonary blastoma and pulmonary sarcomas in children. The lung dose, however, has never been calculated despite the potential risk of lung toxicity from treatment. In this work the dosimetry has been calculated in target tissue and lung for pediatric phantoms. Pleural cavities were modeled in the Monte Carlo code MCNP within the pediatric MIRD phantoms. Both the depth-dose curves in the pleural lining and into the lung as well as 3D dose distributions were calculated for either homogeneous or inhomogeneous (32)P activity distributions. Dose-volume histograms for the lung tissue and isodose graphs were generated. The results for the 2D depth-dose curve to the pleural lining and tumor around the pleural cavity correspond well with the point kernel model-based recommendations. With a 2 mm thick pleural lining, one-third of the lung parenchyma volume gets a dose more than 30 Gy (V(30)) for 340 MBq (32)P in a 10 year old. This is close to lung tolerance. Younger children will receive a larger dose to the lung when the lung density remains equal to the adult value; the V(30) relative lung volume for a 5 year old is 35% at an activity of 256 MBq and for a 1 year old 165 MBq yields a V(30) of 43%. At higher densities of the lung tissue V(30) stays below 32%. All activities yield a therapeutic dose of at least 225 Gy in the pleural lining. With a more normal pleural lining thickness (0.5 mm instead of 2 mm) the injected activities will have to be reduced by a factor 5 to obtain tolerable lung doses in pediatric patients. Previous dosimetry recommendations for the adult apply well down to lung surface areas of 400 cm(2). Monte Carlo dosimetry quantitates the three-dimensional dose distribution, providing a better insight into the maximum tolerable activity for this therapy.

  10. Phosphorus use efficiency by cotton measured through 32P isotope technique

    NASA Astrophysics Data System (ADS)

    Marcante, N. C.; Muraoka, T.; Camacho, M. A.; César, F. R. C. F.; Bruno, I. P.

    2012-04-01

    Deficiency of phosphorus (P) is the major limitation to agricultural production in the Brazilian Savannah (Cerrado), which is naturally poor in this nutrient. Most of the P applied by fertilizer in Cerrado soils are converted into low solubility forms and can not be easily absorbed by plants. This occurs for characteristics of adsorption, conditioned by the predominance of low pH and aluminum and iron oxides in the clay fraction. The development of genotypes and cultivars with greater capacity to grow up in soils with low P availability ('phosphorus efficiency') is interesting to improve the agriculture in these areas in a sustainable way. Cotton (Gossypium spp.) is the main product for the fibers used nationally and globally in the textile chain. This study aim was to evaluate the efficiency of absorption and utilization of P by cotton cultivars/genotypes grown in Cerrado soil by the isotopic dilution technique. The soil classified as Ultisols, was labeled with the radioisotope 32P.The experiment was conducted in a greenhouse in a completely randomized design factorial 2 x 17. Factors were considered two levels of P (insufficient = 20 mg kg-1 and sufficient = 120 mg kg-1) and 17 genetic materials of cotton recommended for Cerrado region. Phosphorus levels influenced significantly the shoots dry matter production, the P content and accumulation, the 32P specific activity, the L value and L value less seed cotton P by cultivars and genotypes. The hierarchical clustering analysis used to verify the similarities between the cultivars and genotypes of cotton, classified them into internally homogeneous groups and heterogeneous between different groups. Cultivars FMT 523, FM 910 and CNPA GO 2043 were the most responsive to phosphate fertilizer in sufficient level of P, while the genotype Barbadense 01 and cultivars FM 966LL, IPR Jataí, BRS Aroeira and BRS Buriti were most efficient absorbing P in soils with insufficient level.

  11. Comparison of /sup 32/P therapy and sequential hemibody irradiation (HBI) for bony metastases as methods of whole body irradiation

    SciTech Connect

    Aziz, H.; Choi, K.; Sohn, C.; Yaes, R.; Rotman, M.

    1986-06-01

    We report a retrospective study of 15 patients with prostate carcinoma and diffuse bone metastases treated with sodium /sup 32/P for palliation of pain at Downstate Medical Center and Kings County Hospital from 1973 to 1978. The response rates, duration of response, and toxicities are compared with those of other series of patients treated with /sup 32/P and with sequential hemibody irradiation. The response rates and duration of response are similar with both modalities ranging from 58 to 95% with a duration of 3.3 to 6 months with /sup 32/P and from 75 to 86% with a median duration of 5.5 months with hemibody irradiation. There are significant differences in the patterns of response and in the toxicities of the two treatment methods. Both methods cause significant bone marrow depression. Acute radiation syndrome, radiation pneumonitis, and alopecia are seen with sequential hemibody irradiation and not with /sup 32/P, but their incidence can be reduced by careful treatment planning. Hemibody irradiation can provide pain relief within 24 to 48 h, while /sup 32/P may produce an initial exacerbation of pain. Lower hemibody irradiation alone is less toxic than either upper hemibody irradiation or /sup 32/P treatment.

  12. Effects of intraarticular 32P colloid in the treatment of hemophilic synovitis of the knee: A short term clinical study

    PubMed Central

    Zhang, Wen-Qiang; Han, Shao-Qin; Yuan, Zhen; He, Ye-Teng; Zhang, Hu; Zhang, Ming

    2016-01-01

    Background: Chronic synovitis is a consequence of recurrent intraarticular hemorrhage in patients with hemophilia. Eventually, synovitis leads to degeneration of the articular cartilage, with serious consequences that impact the quality-of-life in hemophiliacs. The aim of our study was to investigate the short term clinical effects of intraarticular injection of the radionuclide preparation32P colloid (32P-labelled colloidal chromic phosphate suspension) on recurrent intraarticular hemorrhages in patients with hemophilic synovitis of the knee. Materials and Methods: Patients who met the inclusion criteria (n = 22) were enrolled in an open-label study between October 2011 and September 2012.32P colloid was injected into the knee joint and patients were followed up over 6 months after treatment. Hemorrhage frequency, visual analog scale pain score, hospital for special surgery knee score, knee circumference, upper knee circumference, knee diameter, and knee range of motion (ROM) were compared before and after treatment with intraarticular32P colloid injection. Results: In 24 knees evaluated in 22 participating patients, there was a significant reduction in the number of hemorrhages after32P colloid treatment, along with significant pain relief. However, there were no statistically significant changes in the degree of joint swelling, degree of muscle atrophy and knee ROM between the pre and post treatment evaluations. Conclusion: The frequency of joint hemorrhage in patients with hemophilic knee synovitis can be significantly reduced and local symptoms can be improved in the short term by intraarticular injection of32P colloid. PMID:26955177

  13. Hair 32P measurement for body dose mapping in non-fatal exposures to fast neutrons.

    PubMed

    Mianji, Fereidoun A; Jafari, Sheyda; Zaryouni, Saiedeh; Hajizadeh, Bardia

    2015-03-01

    Dosimetry bioassay methods are the backbone of a personal dosimetry in criticality accidents. Although methods like hair dosimetry and the use of activation foils (e.g., (32)S) have been employed for decades, capabilities of different techniques, effects of hair type and neutron spectrum on the dose response, sensitivity and uncertainties of different techniques, etc., need more investigations. For this reason, the use of the (32)S(n,p)(32)P reaction and hair samples for estimating non-fatal doses from fast neutrons was studied. The experiments were carried out with the hair samples attached on a RANDO phantom in a Cf-252 neutron field, in the dose range of about 0.05-1.15 Gy. In addition, the adequate post-accident preparation for hair samples including optimum conditioning and timing were investigated. Experimental results prove the good sensitivity and merit of the method for neutron quantification in the mentioned dose range for which other bioassay methods are of poor resolution and sensitivity. A rough estimation of the dose-response curve for Iranian hair was also derived.

  14. 32P-postlabeling assay for the quantification of the major platinum-DNA adducts.

    PubMed

    Pluim, D; Maliepaard, M; van Waardenburg, R C; Beijnen, J H; Schellens, J H

    1999-11-01

    To allow more sensitive, selective, and routine analyses of platinum(Pt)-GG and -AG intrastrand cross-links we have significantly improved our quantitative (32)P-postlabeling assay (M. J. P. Welters et al. Carcinogenesis 18, 1767-1774, 1997). Instead of off-line scintillation counting we introduced an on-line flow radioisotope detector into the HPLC system. Furthermore, the isolation protocol for the adducts was significantly modified and optimized to reduce interfering background peaks that prevented quantification of low levels of the cisplatin-DNA adducts in white blood cells obtained from patients. Reduction of background signals was obtained by boiling the samples, followed by phenol/chloroform/isoamylethanol extraction after the DNA digestion step. The labeling efficiency for the adducts was increased by 40% by using Na-formate instead of NH(4)-formate for elution of the adducts from the strong cation-exchange columns. Finally, a calibration curve and quality controls were implemented. The labeling efficiencies were not different between the dinucleotides. The between- and within-run precision for the Pt-GG and Pt-AG adducts measured at the lower limit of quantification of 87 and 53 amol/microg DNA, respectively, was less than 20% CV. The adducts were stable in DNA stored for a 2-month time period at -80 degrees C. The assay is now routinely used for high-precision analyses of patient and cell line samples containing very low adduct levels. Copyright 1999 Academic Press.

  15. Synoviorthesis with colloidal /sup 32/P chromic phosphate for hemophilic arthropathy: clinical follow-up

    SciTech Connect

    Rivard, G.E.; Girard, M.; Lamarre, C.; Jutras, M.; Danais, S.; Guay, J.P.; Belanger, R.D.

    1985-11-01

    Thirty-one synoviortheses were performed in 22 joints of 14 hemophilic patients (aged 12 to 28 years) with chronic synovitis and for whom conventional treatments were considered ineffective. Except for patients with inhibitors, conventional treatments included three to six months of adequate prophylactic therapy with the missing coagulation factors, intensive physiotherapy and, when indicated, antiinflammatory agents and orthosis. Colloidal /sup 32/P chromic phosphate was injected intraarticularly in doses of 1.0 mCi for knees and of 0.5 mCi for the other joints. Time of follow-up ranged from two to five years. Frequency and importance of bleeding decreased in all patients. Effect on range of motion was best in knees. In elbows, flexion-extension was improved in four cases, unchanged in five and decreased in one; pronation-supination was decreased in four cases. The results of 13 synoviortheses in four hemophilic patients with high titer factor VIII inhibitors were comparable to those in hemophiliacs with no inhibitors. However, in three of the four patients synoviorthesis had to be repeated after two to four years for recurrence of synovitis. Extraarticular escape of radioactivity was monitored 62 times for 17 synoviortheses in 12 patients; extraarticular counts never exceeded 4% of the intraarticular counts. Chromosome aberrations were found not to be increased after treatment in the seven patients in whom adequate analysis could be done.

  16. [32P]2-iodo-N6-methyl-(N)-methanocarba-2′-deoxyadenosine-3′,5′-bisphosphate ([32P]MRS2500), a novel radioligand for quantification of native P2Y1 receptors

    PubMed Central

    Houston, Dayle; Ohno, Michihiro; Nicholas, Robert A; Jacobson, Kenneth A; Harden, T Kendall

    2005-01-01

    Analysis of the P2Y family of nucleotide-activated G-protein-coupled receptors has been compromised by the lack of selective high-affinity, high-specific-radioactivity radioligands. We have pursued quantification of the P2Y1 receptor through the development of a series of selective P2Y1 receptor antagonists. Recently, we synthesized 2-iodo-N6-methyl-(N)-methanocarba-2′-deoxyadenosine 3′,5′-bisphosphate (MRS2500), a selective, competitive antagonist that exhibits a Ki of 0.8 nM in competition-binding assays with [3H]MRS2279. A 3′-monophosphate precursor molecule, MRS2608, was radiolabeled at the 5′ position with 32P using polynucleotide kinase and [γ32P]ATP to yield [32P]MRS2500. [32P]MRS2500 bound selectively to Sf9 insect cell membranes expressing the human P2Y1 receptor (Sf9-P2Y1), but did not detectably bind membranes expressing other P2Y receptors. P2Y1 receptor binding to [32P]MRS2500 was saturable with a KD of 1.2 nM. Agonists and antagonists of the P2Y1 receptor inhibited [32P]MRS2500 binding in Sf9-P2Y1 membranes with values in agreement with those observed in functional assays of the P2Y1 receptor. A high-affinity binding site for [32P]MRS2500 (KD=0.33 nM) was identified in rat brain, which exhibited the pharmacological selectivity of the P2Y1 receptor. Distribution of this binding site varied among rat tissues, with the highest amount of binding appearing in lung, liver, and brain. Among brain regions, distribution of the [32P]MRS2500 binding site varied by six-fold, with the highest and lowest amounts of sites detected in cerebellum and cortex, respectively. Taken together, these data illustrate the synthesis and characterization of a novel P2Y1 receptor radioligand and its utility for examining P2Y1 receptor expression in native mammalian tissues. PMID:16299552

  17. 32P-incorporation PCR for the detection of rearrangements at the TCR-gamma locus.

    PubMed

    Short, M A; Evans, P A; Shiach, C R; Jack, A; Richards, S; Morgan, G J

    1996-03-01

    We have adapted and developed a PCR (polymerase chain reaction)-based technique for the T-cell receptor (TCR)-gamma chain gene, which has subsequently been used for routine diagnosis. Variable-region oligonucleotide primers were chosen from subgroups I and II, and the joining region primer was from the J2 segment. The primers were used to perform a 32P-incorporation PCR, and the products were then separated on an 8% denaturing polyacrylamide gel. In our hands, this technique is more reliable than cold methods, when separation is performed on either agarose or nondenaturing polyacrylamide. The radioactive technique was used to look at 102 T-cell proliferations, of which eight of eight T-acute lymphoblastic leukemia (ALL), 24 of 34 T-non-Hodgkin's leukemia (NHL), and 35 of 60 large granular lymphocyte (LGL) expansions were clonal. Of 122 B-cell proliferations investigated, including 72 cases of B-cell lineage ALL, 36 demonstrated a T-cell rearrangement (33 ALLs and three myelomas). Samples from nonlymphoid tumors were tested and produced a normal distribution ladder of PCR products after autoradiography, a pattern also observed with antenatal and preoperative patients. The radiolabel-incorporation method detected an abnormal pattern of a ladder with prominent dark bands in 29 of 122 B-cell and 27 of 102 T-cell cases and in 0 of 49 of the nonlymphoid and normal samples. The abnormal banding patterns obtained in a proportion of the B- and T-cell cases was not readily discernible by nondenaturing-acrylamide or agarose-separation methods.

  18. Phosphorous retention in a remediated stream - evaluation of a 32P tracer experiment

    NASA Astrophysics Data System (ADS)

    Riml, Joakim; Morén, Ida; Wörman, Anders

    2017-04-01

    The increased attention to surface water quality problems together with the revealed importance of the stream water -hyporheic zone system for solute retention has highlighted the potential for surface water systems to mitigate solute export to downstream recipients. As a consequence, the number of stream restoration projects during the last decades has increased significantly. However, to be able to design remediation measures as well as to assess the effectiveness of implemented measures, quantitative knowledge of the hydrodynamic (substance independent) and the biogeochemical processes (substance dependent) retaining the solute along the transport pathway is needed. In this work, we present the findings from a simultaneous injection of tritiated water (3H20) and phosphate (32PO4-) with the overall aim to evaluate the effectiveness of remediation actions implemented along a 6 km stretch of a small agricultural stream in Sweden. In contrast to many other tracer tests where different types of proxy substances are used, a key advantage of the study is the use of the substance of environmental interest (in this case phosphorous), which enhances the significance of the results. In addition, the unique radioactive signal from the injected tracer allowed us to distinguish the added phosphorous from other diffuse sources of phosphorous from the surrounding landscape. By using a physically based transport model to evaluate the tracer breakthrough curves at a number of subsequent sampling stations, we were able to contrast the response of different stream reaches both with respect to hydrodynamic and biogeochemical retention. In particular, we found a substantial importance of vegetation on the retention of 32P, when comparing established reaches with dense in-stream vegetation with newly implemented reaches where vegetation was completely absent.

  19. Recoveries of DNA adducts of polycyclic aromatic hydrocarbons in the 32P-postlabelling assay.

    PubMed

    Segerbäck, D; Vodicka, P

    1993-12-01

    The 32P-postlabelling assay for analysis of DNA adducts of chemical carcinogens has been applied in a large number of experimental animal and human studies. Most human studies have dealt with occupational and environmental exposures to polycyclic aromatic hydrocarbons (PAHs). The postlabelling assay does not allow direct chemical identification, and most studies with this method have not been performed in a quantitative way. Very little is therefore known about the identity and absolute levels of adducts, which are important contributors to the process of risk identification and quantitation. In the present study it was, therefore, decided to test some parameters suspected to affect recoveries of adducts in the phosphorylation step of the assay. For this purpose 12 different PAHs were reacted individually and in a mixture with DNA in the presence of a rat liver S9 metabolizing system. Different concentrations of ATP, calcium chloride and polynucleotide kinase were tested using the nuclease P1 enhancement. We found that each factor contributed to adduct recovery and that optimal conditions could be defined. Diluting the modified DNA samples up to 1000 times had little influence on the recoveries of adducts. Comparing the nuclease P1 and the butanol extraction procedures for adduct purification showed that both methods gave similar patterns and levels of major adducts. The absolute recoveries in postlabelling, based on 3H-binding of radiolabelled compounds, were for most of the tested compounds relatively low. The fact that the nuclease P1 and the butanol extraction procedures gave similar recoveries points towards common factor(s) involved in the reduction of the recovered adduct levels. Based on the observed recoveries the conclusion can be drawn that when postlabelling related adducts in human samples the true total adduct levels can be considerably underestimated, even if optimal conditions are used.

  20. Ion-implantation and characterization of 32P-radioactive platinum coils for endovascular treatment of intracranial aneurysms

    NASA Astrophysics Data System (ADS)

    Leblanc, Philippe; Raymond, Jean; Roorda, Sjoerd

    2006-01-01

    We produced and measured over 800 32P-ion-implanted coils for pre-clinical and clinical studies. Platinum coils are intravascular implants most frequently used in the treatment of intracranial aneurysms. This less invasive endovascular approach is safer than conventional surgery, but a frequent drawback is the recurrence of the aneurysm, associated with recanalization, a phenomenon that can be inhibited by the local application of beta radiation. Total coil activities, uniformity, reproducibility and 32P binding to platinum were determined and found to be adequate for this application.

  1. TSH stimulates 32P-labeling of thyroid nuclear HMG 14, a protein associated with actively transcribed chromatin

    SciTech Connect

    Cooper, E.; Palmer, R.J.; Spaulding, S.W.

    1982-04-01

    Thyroid slices were incubated with 32P with or without TSH. 32P-labeling of acid-soluble nuclear proteins was then examined by two-dimensional polyacrylamide gel electrophoresis and autoradiography. We found that TSH enhanced the labeling of the high mobility group protein HMG 14, a protein that is preferentially associated with actively transcribed chromatin. This observation suggests that changes in HMG 14 phosphorylation may be involved in mediating TSH-induced effects on the structure and function of active chromatin.

  2. Simulation of a 32P Sourcewire and a 90Sr/90Y Sourcetrain Using MCNP4b and EGS4

    NASA Astrophysics Data System (ADS)

    Todorovic, M.; Cremers, F.; Albers, D.; Schmidt, R.

    The two simulated sources, the 32P sourcewire and the 90Sr/90Y sourcetrain, are parts of catheter-based systems developed for vascular brachytherapy. Percutaneous transluminal coronary angioplasty (PTCA) is a widely accepted therapy for symptomatic coronary disease, but within the first 6 months often restenosis occur. The intracoronary brachytherapy is a new method to help avoiding restenosis [1].

  3. Dosimetric comparison of {sup 90}Y, {sup 32}P, and {sup 186}Re radiocolloids in craniopharyngioma treatments

    SciTech Connect

    Sadeghi, Mahdi; Karimi, Elham; Hosseini, S. Hamed

    2009-11-15

    Purpose: In the radionuclide treatment of some forms of brain tumors such as craniopharyngiomas, the selection of the appropriate radionuclide for therapy is a key element in treatment planning. The aim was to study the influence by considering the beta-emitter radionuclide dose rate in an intracranial cyst. Methods: Dosimetry was performed using the MCNP4C radiation transport code. Analytical dosimetry was additionally performed using the Loevinger and the Berger formulas in the MATLAB software. Each result was compared under identical conditions. The advantages and disadvantages of using {sup 90}Y versus {sup 32}P and {sup 186}Re were investigated. Results: The dose rate at the inner surface of the cyst wall was estimated to be 400 mGy/h for a 1 MBq/ml concentration of {sup 90}Y. Under identical conditions of treatment, the corresponding dose rates were 300 mGy/h for {sup 32}P and 160 mGy/h for {sup 186}Re. For a well-defined cyst radius and identical wall thickness, higher dose rates resulted for {sup 90}Y. Conclusions: To achieve the same radiological burden, the required amount of physical activity of injectable solution is lower for {sup 32}P. This is found to be a consequence of both the radionuclide physical half-life and the pattern of energy deposition from the emitted radiation. According to the half-life and dose-rate results, {sup 90}Y would be a good substitute for {sup 32}P.

  4. Patient-specific dosimetry for intracavitary 32P-chromic phosphate colloid therapy of cystic brain tumours.

    PubMed

    Denis-Bacelar, Ana M; Romanchikova, Marina; Chittenden, Sarah; Saran, Frank H; Mandeville, Henry; Du, Yong; Flux, Glenn D

    2013-10-01

    (32)P-chromic phosphate colloid treatments of astrocytoma and craniopharyngioma cystic brain tumours in paediatric patients are conventionally based on a sphere model under the assumption of uniform uptake. The aims of this study were to determine the distribution of the absorbed dose delivered by (32)P on a patient-specific basis and to evaluate the accuracy with which this can be predicted from a pretherapy administration of (99m)Tc-Sn colloid. Three patients were treated with (32)P-chromic phosphate colloid following (99m)Tc-Sn colloid administrations. Convolution dosimetry was performed using pretherapy and posttherapy sequential SPECT imaging, and verified with EGSnrc Monte Carlo radiation transport simulations. Mean absorbed doses to the cyst wall and dose-volume histograms were also calculated and compared with those obtained by the sphere model approach. Highly nonuniform uptake distributions of both the (99m)Tc and (32)P colloids were observed and characterized by dose-volume histograms to the cyst wall. Mean absorbed doses delivered to the cyst wall, obtained with the convolution method, were on average 21 % (SD 18 %) and 50 % (SD 30 %) lower than those predicted by the (99m)Tc distribution and the uniform assumption of the sphere model, respectively. Absorbed doses delivered to the cyst wall by (32)P are more accurately predicted from image-based patient-specific convolution dosimetry than from simple sphere models. These results indicate the necessity to perform personalized treatment planning and verification for intracavitary irradiation of cystic brain tumours treated with radiocolloids. Patient-specific dosimetry can be used to guide the frequency and levels of repeated administrations and would facilitate data collection and comparison to support the multicentre trials necessary to progress this therapy.

  5. Determination of surface dose rate of indigenous (32)P patch brachytherapy source by experimental and Monte Carlo methods.

    PubMed

    Kumar, Sudhir; Srinivasan, P; Sharma, S D; Saxena, Sanjay Kumar; Bakshi, A K; Dash, Ashutosh; Babu, D A R; Sharma, D N

    2015-09-01

    Isotope production and Application Division of Bhabha Atomic Research Center developed (32)P patch sources for treatment of superficial tumors. Surface dose rate of a newly developed (32)P patch source of nominal diameter 25 mm was measured experimentally using standard extrapolation ionization chamber and Gafchromic EBT film. Monte Carlo model of the (32)P patch source along with the extrapolation chamber was also developed to estimate the surface dose rates from these sources. The surface dose rates to tissue (cGy/min) measured using extrapolation chamber and radiochromic films are 82.03±4.18 (k=2) and 79.13±2.53 (k=2) respectively. The two values of the surface dose rates measured using the two independent experimental methods are in good agreement to each other within a variation of 3.5%. The surface dose rate to tissue (cGy/min) estimated using the MCNP Monte Carlo code works out to be 77.78±1.16 (k=2). The maximum deviation between the surface dose rates to tissue obtained by Monte Carlo and the extrapolation chamber method is 5.2% whereas the difference between the surface dose rates obtained by radiochromic film measurement and the Monte Carlo simulation is 1.7%. The three values of the surface dose rates of the (32)P patch source obtained by three independent methods are in good agreement to one another within the uncertainties associated with their measurements and calculation. This work has demonstrated that MCNP based electron transport simulations are accurate enough for determining the dosimetry parameters of the indigenously developed (32)P patch sources for contact brachytherapy applications.

  6. Synoviorthesis with 32P-colloidal chromic phosphate in rheumatoid arthritis--clinical, histopathologic and arthrographic changes.

    PubMed

    Onetti, C M; Gutiérrez, E; Hliba, E; Aguirre, C R

    1982-01-01

    Synoviorthesis was performed in 217 joints from 111 patients suffering from different stages of rheumatoid arthritis (RA). 32P-colloidal chromic phosphate was employed, with an average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as average dose from 6 mCi for large joints (knees) to 0.3 mCi for small peripheral joints such as the MCP or PIP joints. Satisfactory clinical results were observed in 84% of the cases and no significant side effects resulted after a follow-up period from 1 to 10 years. Striking effects after treatment were observed through histopathological studies (light and electron microscopy) and the use of contrast arthrography. We concluded that radioactive synovectomy with 32P-chromate is a very useful method for the local treatment of RA.

  7. Marrow toxicity of 33P-versus 32P-orthophosphate: implications for therapy of bone pain and bone metastases.

    PubMed

    Goddu, S M; Bishayee, A; Bouchet, L G; Bolch, W E; Rao, D V; Howell, R W

    2000-05-01

    Several bone-seeking radiopharmaceuticals, such as 32P-orthophosphate, 89Sr-chloride, 186Re-1,1 hydroxyethylidene diphosphonate (HEDP), and 153Sm-ethylene diamine tetramethylene phosphonic acid (EDTMP), have been used to treat bone pain. The major limiting factor with this modality is bone marrow toxicity, which arises from the penetrating nature of the high-energy beta particles emitted by the radionuclides. It has been hypothesized that marrow toxicity can be reduced while maintaining therapeutic efficacy by using radionuclides that emit short-range beta particles or conversion electrons. In view of the significant clinical experience with 32P-orthophosphate, and the similarity in pain relief afforded by 32P-orthophosphate and 89Sr-chloride, this hypothesis is examined in this study using 32P- and 33P-orthophosphate in a mouse femur model. Survival of granulocyte macrophage colony-forming cells (GM-CFCs) in femoral marrow was used as a biologic dosimeter for bone marrow. 32P- and 33P-orthophosphate were administered intravenously, and GM-CFC survival was determined as a function of time after injection and, at the nadir, as a function of injected activity. The kinetics of radioactivity in the marrow, muscle, and femoral bone were also determined. The biologic dosimeter was calibrated by assessing GM-CFC survival at its nadir after chronic irradiation of Swiss Webster mice with exponentially decreasing dose rates of gamma rays (relative biologic effectiveness equivalent to that of beta particles) from a low-dose rate 137Cs irradiator. Dose-rate decrease half-times (Td) (time required for 137Cs gamma ray dose rate to decrease by one half) of 62, 255, and 425 h and infinity were used to simulate the dose rate patterns delivered by the radiopharmaceuticals as dictated by their effective clearance half-times from the mouse femurs. These data were used to experimentally determine the mean absorbed dose to the femoral marrow per unit injected activity. Finally, a

  8. Detection of human cytomegalovirus by slot-blot hybridization assay employing oligo-primed /sup 32/P-labelled probe

    SciTech Connect

    Agha, S.A.; Coleman, J.C.; Selwyn, S.; Mahmound, L.A.; Abd-Elaal, A.M.; Archard, L.C.

    1988-12-01

    A /sup 32/P-labelled Hind III-0 DNA fragment (nine Kilobases; Kb) from human cytomegalovirus AD-169 (HCMV) was used in slot-blot hybridization assay for the detection of HCMV in clinical samples. The results obtained with DNA hybridization assay (DNA HA) were compared with virus isolation using conventional tube cell culture (CTC) and centrifugation vial culture (CVC), immunofluorescence (IF), and complement fixation test (CFT). Of 15 CTC-positive samples, 13 were positive with DNA HA (sensitivity 86.7%). Also, 14 additional samples were DNA HA-positive but CTC-negative. CVC and/or IF confirmed the diagnosis in nine of 14; the remaining five samples were from three patients who showed fourfold rising antibody titre by CFT. Although DNA HA using /sup 32/P-labelled probes is relatively cumbersome and expensive, it is a valuable test for quantitation of viral shedding in patients with HCMV infections who may benefit from antiviral therapy.

  9. Determination of natural 32P and 33P in rainwater, marine particles and plankton by low-level beta counting

    NASA Astrophysics Data System (ADS)

    Waser, N. A.; Fleer, A. P.; Hammar, T. R.; Buesseler, K. O.; Bacon, M. P.

    1994-01-01

    Methods were developed for determining the natural levels of 32P and 33P in samples of rainwater, marine particulate matter and plankton. Preconcentration from rainwater consists of the extraction of radiophosphorus on alumina. Filtration of large volumes of seawater is required to obtain sufficient quantities of suspended particulate matter. The radiochemical scheme, which consists of a series of precipitations and a column separation, results in a pure source containing the two beta emitters. The activities of 32P and 33P in the source are determined separately by an external absorber method on low-level beta counters. The yields are monitored by the addition of stable phosphate in rainwater samples and by the natural levels of stable phosphate in marine particulate matter and plankton. The methods allow the specific activities and the activity ratio to be determined with a 10% precision.

  10. Biologic dosimetry of bone marrow: induction of micronuclei in reticulocytes after exposure to 32P and 90Y.

    PubMed

    Lenarczyk, M; Goddu, S M; Rao, D V; Howell, R W

    2001-01-01

    Bone marrow is the dose-limiting organ in targeted radionuclide therapy. Hence, determination of the absorbed dose to bone marrow from incorporated radionuclides is a critical element in treatment planning. This study investigated the potential of the micronucleus assay in peripheral blood reticulocytes (MnRETs) as an in vivo biologic dosimeter for bone marrow. After intravenous administration of 32P-orthophosphate or 90Y-citrate in Swiss Webster mice, DNA damage induced in bone marrow erythroblastoid cells was measured by subsequent scoring of MnRETs in peripheral blood. The response to exponentially decreasing dose rates was calibrated by irradiating animals with external 137Cs-gamma-rays. The gamma-ray dose rate was decreased exponentially, with the dose-rate decrease half-time corresponding to the effective clearance half-time (Te) of the radioactivity from the femoral bone (Te = 64 h for 90Y-citrate and Te = 255 h for 32P-orthophosphate). The maximum MnRETs frequency occurred on the second and third day after injection of 90Y-citrate and 32P-orthophosphate, respectively. The same pattern was observed for exponentially decreasing dose rates of 137Cs-gamma-rays. For each type of exposure, the maximum MnRETs frequency increased in a dose-dependent manner. Using the calibrated dosimeter, the initial dose rates to the marrow per unit of injected activity were 0.0020 cGy/h/kBq and 0.0026 cGy/h/kBq for 32P-orthophosphate and 90Y-citrate, respectively. Micronuclei in peripheral blood reticulocytes can be used as a noninvasive biologic dosimeter for measuring absorbed dose rate and absorbed dose to bone marrow from incorporated radionuclides.

  11. (32)P measurment of urine samples and internal dose assessment for radiation workers in life science laboratories.

    PubMed

    Yoon, S; Pak, M-J; Park, S; Yoo, J; Ha, W-H; Jang, H-K; Kim, J K

    2014-12-01

    (32)P measurements of urine samples and internal dose assessments were conducted for workers in life science laboratories. A procedure for sample pre-treatment was established and validation was performed to exclude interference and to detect (32)P levels accurately. The detection conditions for Cherenkov radiation were evaluated and the accuracy of Cherenkov radiation measurements validated. The analytical and measurement procedures were applied to urine samples collected from 11 workers from life sciences laboratories. The results of the measurements generally indicated very low background radiation levels, but daily urine samples from two workers were above the minimum detectable activity. The (32)P concentrations for two of the workers were 29.3  ±  10.4 Bq•d(-1) and 24.1  ±  11.8 Bq•d(-1), respectively, at intake levels of 4.12 kBq and 2.61 kBq. The effective doses for these two workers were 4.6 μSv and 2.9 μSv. Overall, the results indicate very low levels of radioactivity, except for cases related to specific working conditions.

  12. Bioevaluation study of 32P-CP-PLLA particle brachytherapy in a rabbit VX2 lung tumor model.

    PubMed

    Xu, Yu ping; Yang, Min; Pan, Dong hui; Wang, Li zhen; Liu, Lu; Huang, Peilin; Shao, Guoqiang

    2012-04-01

    The purpose of this study was to investigate the therapy effects of intratumoral administration of (32)P-CP-PLLA particles in a rabbit VX2 lung tumor model. 16 rabbits with tumors were randomly divided into 4 groups. 4 rabbits served as untreated controls, and others received intratumoral administration of (32)P-CP-PLLA particles with CT guidance. The total radioactivities in treated groups were as follows: a low activity was 93 MBq (n=4) (group 1), a medium activity was 185 MBq (n=4) (group 2) and a high activity was 370 MBq (n=4) (group 3). Brachytherapy treated VX2 tumors underwent (18)F-FDG PET/CT at 0 day, 3 day, 7 day and 14 day postinjection. In control group, (18)F-FDG PET/CT images were acquired at the same time points but without any treatment. Bremsstrahlung SPECT images were performed at 14 days after intratumoral brachytherapy in treated groups. After Bremsstrahlung SPECT and last (18)F-FDG PET/CT imagings, the rabbits were euthanized and the tumors were removed for histological examination. Bremsstrahlung SPECT images study indicated that there was no leakage of (32)P out of the injection site at 14 days after treatment. Compared with the control, the tumor volumes in treated groups significantly decreased, and (32)P-CP-PLLA particle produced a reduction in maximum or mean SUV of VX2 tumor (p<0.05). The percentage changes in maximum and mean SUV gradually decreased in group 1 and group 2 from day 3 to day 14 (p<0.05). A transient increase in (18)F-FDG accumulation at group 3 occurred due to the inflammatory reaction elements. Activity dependence was seen in HE and PCNA staining after 14 days treatment among three treated groups (p<0.05). Our data suggested that (32)P-CP-PLLA particle localized on the injecting sites. This novel brachytherapy device efficiently suppressed the growth of the VX2 tumors implanted in the rabbit. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Two methods that facilitate autoradiography of small /sup 32/P-labeled DNA fragments following electrophoresis in agarose gels

    SciTech Connect

    Cockerill, P.N.

    1988-02-01

    Two methods which permit detection by autoradiography of small /sup 32/P-labeled DNA fragments resolved by agarose gel electrophoresis are described. Agarose gel electrophoresis poses problems for autoradiography as (i) the gels are normally too thick to allow autoradiography without being dried first, and (ii) fragments of DNA of 1000 bp or less in length are readily lost during drying. In this study DNA fragments as small as 121 bp have been retained in agarose gels upon drying. This has been achieved by either (i) first fixing the DNA with the cationic detergent cetyltrimethylammonium bromide, or (ii) drying the agarose gels onto Zeta-Probe charge-modified membranes.

  14. Multiple DNA adducts in lymphocytes of smokers and nonsmokers determined by sup 32 P-postlabeling analysis

    SciTech Connect

    Jahnke, G.D.; Thompson, C.L.; Walker, M.P.; Gallagher, J.E.; Lucier, G.W.

    1990-01-01

    Identification of DNA adducts in peripheral lymphocytes could serve as a means of monitoring human exposure to potential genotoxic agents. In the study, DNA from peripheral lymphocytes of smokers and nonsmokers was examined for adducts by the P1 nuclease {sup 32}P-post-labeling technique. Thin layer chromatography (TLC) maps from both groups revealed multiple DNA adducts which ranged from no adducts for one individual to six adducts for a different individual. The total DNA adduct concentrations were approximately one adduct in 10 to the seventh-10 to the eighth power normal nucleotides. Comparison of the adduct TLC profiles revealed individual variation in both pattern and level of DNA adducts. The type and amount of adduct was not influenced by smoking history and remained unchanged in four out of six subjects who were resampled after a one month interval. One adduct detected in lymphocyte DNA co-migrated on TLC with an adduct derived by in vitro incubation of lymphocytes with benzo(a)pyrene (B(a)P). The 3H-nucloside values were consistent with values obtained by {sup 32}P-postlabeling of the same sample (correlation coefficient of 0.88). No relationship was apparent between the capacity of lymphocytes to form a (3H)-B(a)P-derived adduct in vitro and the concentration of the adduct, or total adducts present in untreated lymphocytes.

  15. Factors affecting phosphate uptake by Aerobacter aerogenes in a system relating cell numbers to 32P uptake.

    PubMed

    White, L A; MacLeod, R A

    1971-03-01

    The uptake of phosphate, from media limited in this ion, by resting cells of Aerobacter aerogenes has been investigated and shown to be dependent upon several factors. An incubation medium composed of 10(-2)m K(+), 5 x 10(-3)m Mg(2+), 1 mg of glucose per ml, and 1 muCi of (32)PO(4) (3-) per ml, buffered at pH 6.55 with 0.05 mN-2-hydroxyethyl-piperazine-N'-2-ethanesulfonic acid (HEPES), was found to stimulate optimum accumulation of (32)P-orthophosphate. The temperature of incubation, incubation time, the concentration of unlabeled orthophosphate, as well as arsenate, and several metabolic inhibitors were found to affect the accumulation. The labeled cells were collected on a membrane filter, which had been previously boiled in glass-distilled water, for measurement of the radioactivity accumulated. Under optimum conditions, as few as 20,000 cells were capable of accumulating detectable amounts of (32)P-orthophosphate in 1 hr of incubation.

  16. {sup 32}P-postlabeling analysis of DNA adducts in wild perch (Perca fluviatilis) and northern pike (Esox lucius)

    SciTech Connect

    Ericson, G.; Liewenborg, B.; Balk, L.

    1995-12-31

    Several previous studies have demonstrated a correlation between high concentrations of sediment-associated contaminants and elevated levels of aromatic/hydrophobic DNA adduct levels in the liver of benthic fish species. In the present study DNA adducts was analyzed in coastal populations of perch (Perca fluviatilis) and northern pike (Esox lucius). Fish were sampled from four different sites in a gradient from a heavily industrialized area at the Swedish Baltic coast. For comparison, fish were also caught in a reference area with no main industries and comparatively low levels of contaminants of anthropogenic origin. DNA was extracted from liver and several extrahepatic tissues and DNA adducts were analyzed by the nuclease PI version of the {sup 32}P-postlabeling assay. The autoradiograms derived from DNA of fish from the contaminated sites showed several adduct spots not visible on the autoradiograms derived from fish from the reference area. Total adduct levels were significantly elevated in several tissues in fish from contaminated sites compared to the reference area. Species and tissue-specific differences in adduct levels and the use of {sup 32}P-postlabeling analysis of DNA adducts as a biomarker to monitor the presence and effects of genotoxic chemicals in the aquatic environment are discussed.

  17. Increased brain radioactivity by intranasal 32P-labeled siRNA dendriplexes within in situ-forming mucoadhesive gels

    PubMed Central

    Perez, Ana Paula; Mundiña-Weilenmann, Cecilia; Romero, Eder Lilia; Morilla, Maria Jose

    2012-01-01

    Background Molecules taken up by olfactory and trigeminal nerve neurons directly access the brain by the nose-to-brain pathway. In situ-forming mucoadhesive gels would increase the residence time of intranasal material, favoring the nose-to-brain delivery. In this first approach, brain radioactivity after intranasal administration of 32P-small interference RNA (siRNA) complexed with poly(amidoamine) G7 dendrimers (siRNA dendriplexes) within in situ-forming mucoadhesive gels, was determined. Materials 32P-siRNA dendriplexes were incorporated into in situ-forming mucoadhesive gels prepared by blending thermosensitive poloxamer (23% w/w) with mucoadhesive chitosan (1% w/w, PxChi) or carbopol (0.25% w/w, PxBCP). Rheological properties, radiolabel release profile, and local toxicity in rat nasal mucosa were determined. The best-suited formulation was intranasally administered to rats, and blood absorption and brain distribution of radioactivity were measured. Results The gelation temperature of both formulations was 23°C. The PxChi liquid showed non-Newtonian pseudoplastic behavior of high consistency and difficult manipulation, and the gel retained 100% of radiolabel after 150 minutes. The PxCBP liquid showed a Newtonian behavior of low viscosity and easy manipulation, while in the gel phase showed apparent viscosity similar to that of the mucus but higher than that of aqueous solution. The gel released 35% of radiolabel and the released material showed silencing activity in vitro. Three intranasal doses of dendriplexes in PxCBP gel did not damage the rat nasal mucosa. A combination of 32P-siRNA complexation with dendrimers, incorporation of the dendriplexes into PxCBP gel, and administration of two intranasal doses was necessary to achieve higher brain radioactivity than that achieved by intravenous dendriplexes or intranasal naked siRNA. Conclusion The increased radioactivity within the olfactory bulb suggested that the combination above mentioned favored the

  18. On the origins and development of the (32)P-postlabelling assay for carcinogen-DNA adducts.

    PubMed

    Phillips, David H

    2013-06-28

    The (32)P-postlabelling method for the analysis of carcinogen-DNA adducts originated 30years ago from Baylor College of Medicine in Houston and was the work of a team comprised of Kurt and Erica Randerath, Ramesh Gupta and Vijay Reddy. With subsequent modifications and developments, it has become a highly sensitive and versatile method for the detection of DNA adducts that has been applied in a wide range of human, animal and in vitro studies. These include monitoring human exposure to environmental and occupational carcinogens, investigating genotoxicity of chemicals, elucidating pathways of metabolic activation of carcinogens, mechanistic studies of DNA repair, analysing the genotoxicity of complex mixtures and in ecotoxicology studies. Its use has been instrumental in providing new clues to the aetiology of some cancers and in identifying a new human carcinogen. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  19. DNA damage induced in mouse tissues by organic wood preserving waste extracts as assayed by 32P-postlabeling.

    PubMed

    Randerath, E; Zhou, G D; Donnelly, K C; Safe, S H; Randerath, K

    1996-01-01

    Numerous wood preserving waste (WPW) sites in the United States pose genotoxic hazards. WPWs consist of complex mixtures containing toxic, including genotoxic, compounds which are derived from the preservatives coal tar creosote and pentachlorophenol (PCP) and other polychlorinated aromatics. The genotoxicity of WPW extracts, which has not been tested in mammals, cannot be evaluated on the basis of data for individual components because of possible compound interactions. Therefore, whole extracts need to be assayed. 32P-postlabeling represents a powerful tool to determine DNA adduct formation by complex genotoxic mixtures, such as cigarette smoke, diesel exhaust, and coke oven and foundry emissions in experimental animals and humans. In the present study, a mouse bioassay was used in combination with 32P-postlabeling to determine DNA adduct formation induced by hexane/acetone extracts of two samples from a WPW site. Female ICR mice were treated dermally with extract corresponding to 3 mg residue or vehicle control once per day for 2 days and killed 24 h later. Skin, lung, liver, kidney, and heart DNA preparations were assayed by nuclease P1-enhanced postlabeling. Adduct profiles were tissue-specific and displayed a multitude of non-polar DNA adducts with levels amounting to one adduct in 1.6 x 10(6) DNA nucleotides in skin (both extracts) and one adduct in 3.2 x 10(7) or 1.2 x 10(7) DNA nucleotides in liver (extract 1 or extract 2). Based on their chromatographic properties, these adducts appeared largely derived from polycyclic aromatic hydrocarbons (PAHs) present in the extracts. One of the major adducts was identified as the 32P-labeled derivative of the reaction product of 7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-epoxy-7, 8,9,10-tetrahydrobenzo[a]pyrene (BPDE I) with N2 of deoxyguanosine. Total non-polar DNA adduct levels were highest in skin and lung, amounting to 17.4 and 24.0% of the skin values for extracts 1 and 2, respectively, in lung while the

  20. Development of samarium [32P] phosphate colloid for radiosynoviorthesis applications: preparation, biological and preliminary clinical studies experience.

    PubMed

    Prabhakar, G; Sachdev, Satbir S; Umamaheswari, S; Sivaprasad, N; Bhatia, Manohar H; Chaudhari, Pradip R; Solav, Srikant V

    2007-12-01

    A new therapeutic radio colloid for radiosynoviorthesis (RS) applications is reported. The method of preparation involves the reaction of SmCl3 carrier with carrier added [32P]H3PO4 in the presence of gelatin. The pure colloid was recovered by dialysis purification leading to radiochemical yield of around 90%. The radiochemical purity of the pure colloid formulated in isotonic saline was over 98%, for the usage period of 14 days, as assessed by paper chromatography. Ninety percent of colloid particles were in the size of 1-10 microm as evident from the laser diffraction particle size analysis, ideally suitable for the intended end use. Animal studies revealed complete retention of the radio colloid in the rabbit knee joint. The results of clinical trials in humans are satisfactory and encouraging, satisfactory retention of the colloid in the knee joint and negligible leakage into the systemic circulation.

  1. {sup 32}P-postlabeling determination of DNA adducts in the earthworm Lumbricus terrestris exposed to PAH-contaminated soils

    SciTech Connect

    Walsh, P. |; El Adlouni, C.; Mukhopadhyay, M.J.; Nadeau, D.; Poirier, G.G.; Viel, G.

    1995-05-01

    The importance of the search for reliable biomarkers of DNA damage in environmental health assessment is well recognized by the scientific community and regulatory agencies. Among the major biomarkers of DNA damage is the measurement of DNA adducts in target cells or tissues. Up to now, DNA adduct determinations have been directed mostly toward human exposure to toxic substances from the workplace and environment. Moreover, techniques for measuring DNA adducts, and in particular the {sup 32}P-postlabelling technique, presented also the possibility of determining DNA adduct levels in endogenous animal populations exposed to polluted environments as early warning monitors of ecotoxicity. Soil contamination is becoming a major environmental issue. Therefore, numerous contaminated sites must now be remediated to protect human health and to permit new uses of these sites as agricultural, residential, or industrial areas. Fulfillment of this task requires standardized and sensitive bioassays to carry out site evaluations and to establish scientifically defensible soil quality criteria. To that effect, the earthworm appears to be one of the best organisms for use in soil toxicity evaluation. Earthworms are probably the most relevant soil species, representing 60 to 80% of the total animal biomass in soil. Present soil bioassays focus mostly on plant species with end points like seed germination, root elongation, seedling growth and seedling emergence, and on acute toxicity evaluation (re: LC 50) on the earthworm Eisenia fetida. As yet, a standardized soil invertebrate test for teratogenic or mutagenic end points has not been developed. In this paper, we report the feasibility of DNA adduct determination by {sup 32}P-postlabelling in the earthworm Lumbricus terrestris as a way to detect the presence of genotoxic substances in soils. 20 refs., 1 fig., 1 tab.

  2. /sup 32/P-postlabeling assay for carcinogen-DNA adducts: nuclease P/sub 1/-mediated enhancement of its sensitivity and applications

    SciTech Connect

    Reddy, M.V.; Randerath, K.

    1987-12-01

    Exceedingly sensitive assays are required for the detection of DNA adducts formed in humans exposed to low levels of environmental genotoxicants and therapeutic drugs. A /sup 32/P-postlabeling procedure for detection and quantitation of aromatic carcinogen-DNA lesions with a sensitivity limit of 1 adduct in 10/sup 7/ to 10/sup 8/ nucleotides has been described previously. In the standard procedure, DNA is enzymatically digested to 3'-phosphorylated normal and adducted mononucleotides, which are /sup 32/P-labeled at their 5'-hydroxyl groups by T4 polynucleotide kinase-catalyzed (/sup 32/P) phosphate transfer from (..gamma..-/sup 32/P)ATP. /sup 32/P-labeled derivatives are resolved by TLC, detected by autoradiography, and quantitated by counting. This assay has been recently utilized for the determination and partial characterization of DNA adducts formed in somatic and reproductive tissues of rats given the clinically used anticancer drug, mitomycin C. The drug exhibits similar levels of covalent binding to DNA in most tissues. Further studies have revealed that adducted nucleotides are primarily guanine derivatives that are resistant to 3'-dephosphorylation by Penicillium citrinum nuclease P/sub 1/. The latter observation has been utilized to enhance the /sup 32/P-assay's sensitivity to 1 adduct in 10/sup 10/ nucleotides for a 10-..mu..g DNA sample by postincubation of DNA digests with nuclease P/sup 1/ before /sup 32/P-labeling. The new assay has also shown utility in the analysis of very low levels of age- and tissue-related DNA modifications, which might arise from dietary or endogenous compounds, in untreated rats and humans.

  3. Post-Dilatation Intravascular Brachytherapy Trials on Hypercholesterolemic Rabbits Using {sup 32}P-Phosphate Solutions in Angioplasty Balloons

    SciTech Connect

    Walichiewicz, Piotr Wilczek, Krzysztof; Petelenz, Barbara; Jachec, Wojciech; Jochem, Jerzy; Tomasik, Andrzej; Bilski, Pawel; Gaca, Pawel; Banaszczuk, Joanna; Ihnatowicz, Jerzy; Wodniecki, Jan

    2004-01-15

    Response of peripheral arteries to post-dilatation intravascular brachytherapy (IVBT) using {sup 32}P liquid sources was studied in a rabbit model. The applied sources were angioplasty balloons filled with aqueous solutions of Na{sub 2}H{sup 32}PO{sub 4}, NaCl and iodinated contrast. Dose distribution was calibrated by thermoluminescence dosimetry. The uncertainty of in vitro determinations of the activity-dose dependence was {+-} 15-30%. The animal experiments were performed on rabbits with induced hypercholesterolemia. The {sup 32}P sources were introduced into a randomly chosen (left or right) iliac artery, immediately after balloon injury. Due to the low specific activity of the applied sources, the estimated 7-49 Gy doses on the internal artery surface required 30-100 min irradiations. A symmetric, balloon-occluded but non-irradiated artery of the same animal served as control. Radiation effects were evaluated by comparing the thicknesses of various components of irradiated versus untreated artery walls of each animal. The treatment was well tolerated by the animals. The effects of various dose ranges could be distinguished although differences in individual biological reactions were large. Only the 49 Gy dose at 'zero' distance (16 Gy at 1.0 mm from the balloon surface) reduced hypertrophy in every active layer of the artery wall. The cross-sectional intimal thicknesses after 7, 12, 38 and 49 Gy doses were 0.277, 0.219, 0.357 and 0.196 mm{sup 2} respectively, versus 0.114, 0.155, 0.421 and 0.256 mm{sup 2} in controls (p < 0.05). The lowest radiation dose on the intima induced the opposite effect. Edge intimal hyperplasia was not avoided, which agrees with other reports. The edge restenosis and the variability of individual response to identical treatment conditions must be considered as limitations of the post-dilatation IVBT method. Only application of highest irradiation doses was effective. The irradiation dose should be planned and calculated for

  4. DNA adducts induced by the potential human carcinogen cyclopenta[cd]pyrene using [sup 32]P-postlabeling

    SciTech Connect

    Beach, A.C.

    1993-01-01

    Cyclopenta[cd]pyrene (CPP) is a widespread polycyclic aromatic hydrocarbon with potent mutagenic and carcinogenic activity. CPP is of interest for its prevalence in such notable sources of human exposure to environmental pollution as automotive/diesel engine exhaust emissions. As DNA adduct formation represents an integral phase in the chemical carcinogenesis process, the following studies were undertaken to determine: (1) the utility [sup 32]P-postlabeling for detecting CPP-DNA adducts; (2) the potency and efficacy of CPP to induce DNA adducts in vivo; and (3) the identity of the ultimate mutagenic intermediate(s) of CPP in vivo. CPP-3,4-epoxide has been suggested as the ultimate mutagenic form of CPP. This epoxide was reacted with DNA and deoxynucleotide induced with those formed by CPP and the base-specificity of those adducts. CPP-3,4-epoxide induced at least four major and three minor DNA adducts at the level of deoxyguanosine. All adducts were similar to those formed by CPP in pilot in vivo studies; therefore the ability of CPP to induce adducts was further evaluated. CPP produced up to four major and seven minor adducts in the rat. CPP-DNA adducts were highest in the lung followed by the heart, WBCs, and liver. CPP was also found to induce adducts in vivo across a five orders of magnitude dosing range i.e. 1 [mu]g/kg-50 mg/kg. CPP adducts induced in vitro were found to increase in the presence of epoxide hydrolase inhibitors suggesting their epoxide origin. This prompted evaluation of the similarity of adducts induced by CPP in vivo with those produced by CPP-3,4-epoxide in vitro. Cochromatographic studies revealed that all four major and at least five of the minor adducts induced by CPP were formed by reaction of CPP-3,4-epoxide with deoxyguanosine; one adduct was clearly oxyadenosine derived. Finally, [sup 32]P-postlabeling was used to detect CPP-adducts in assays with potential for human application.

  5. Demonstration of paternal inheritance of plastids in Picea (Pinaceae). [Hybridization of cloned, sup 32 -P labeled, petunia cpDNA

    SciTech Connect

    Stine, M.

    1988-01-01

    Chloroplast DNA (cpDNA) was purified from Picea glauca, P. pungens, P. engelmannii, and P. omorika, and was digested with several restriction endonucleases. Interspecific restriction fragment length polymorphisms (RFLPs) of cpDNA were identified. The RFLPs were identified as cpDNA by the hybridization of cloned, {sup 32}-P labeled, petunia cpDNA to the polymorphic bands, and by the lack of hybridization of a cloned and labeled mtDNA probe from maize. Chloroplast DNA RFLPs that showed no intraspecific variation when examined across the natural range for each species, were used as markers to follow the inheritance of plastids in interspecific hybrids. The inheritance of plastids was determined for F{sub 1}-hybrids from reciprocal crosses of P. glauca and P. pungens, P. glauca and P. omorika, and F{sub 1}-hybrids of P. engelmannii x pungens. All 31 F{sub 1}-hybrids examined showed the cpDNA genotypes of the pollen parent, or the paternal species.

  6. In vitro incorporation of label from (. gamma. /sup 32/P) ATP into isocitrate lyase of Escherichia coli

    SciTech Connect

    Robertson, F.; Reeves, H.C.

    1986-05-01

    A partially purified sonic extract of an E. coli mutant, constitutive for the glyoxylate by-pass enzymes, was incubated with (..gamma../sup 32/P) ATP in 50 mM MOPS buffer at pH 7.5 containing 2 mM 2-mercaptoethanol, 10 mM MgCl/sub 2/ and 10% glycerol at room temperature for 1 h. Incubation was continued for another hour following the addition of unlabeled ATP. The assay was terminated by the addition of 10 mM EDTA. The assay mixture was then analyzed, by several electrophoretic techniques and subsequent autoradiography, to determine which of the proteins in the extract had incorporated label. Isoelectric focusing was performed at pH 3-10 and pH 4-4.5. Specific enzyme staining of IEF gels revealed that active isocitrate lyase (ICL) co-migrated with a protein band which also was radioactive. Immuno blots of these IEF gels, using antibody raised in rabbits against ICL, also demonstrated incorporation of label. SDS-PAGE autoradiograms displayed a labeled protein which co-migrated with purified E. coli ICL, which has a subunit M/sub r/ of 48,000. The mixture was also analyzed by 2-D PAGE which further demonstrated that a labeled protein and authentic purified ICl co-migrate.

  7. A [32P]-NAD+-based method to identify and quantitate long residence time enoyl-ACP reductase inhibitors

    PubMed Central

    Yu, Weixuan; Neckles, Carla; Chang, Andrew; Bommineni, Gopal Reddy; Spagnuolo, Lauren; Zhang, Zhuo; Liu, Nina; Lai, Christina; Truglio, James; Tonge, Peter J.

    2015-01-01

    The classical methods for quantifying drug-target residence time (tR) use loss or regain of enzyme activity in progress curve kinetic assays. However, such methods become imprecise at very long residence times, mitigating the use of alternative strategies. Using the NAD(P)H-dependent FabI enoyl-ACP reductase as a model system, we developed a Penefsky column-based method for direct measurement of tR, where the off-rate of the drug was determined with radiolabeled [adenylate-32P] NAD(P+) cofactor. Twenty-three FabI inhibitors were analyzed and a mathematical model was used to estimate limits to the tR values of each inhibitor based on percent drug-target complex recovery following gel filtration. In general, this method showed good agreement with the classical steady state kinetic methods for compounds with tR values of 10-100 min. In addition, we were able to identify seven long tR inhibitors (100-1500 min) and to accurately determine their tR values. The method was then used to measure tR as a function of temperature, an analysis not previously possible using the standard kinetic approach due to decreased NAD(P)H stability at elevated temperatures. In general, a 4-fold difference in tR was observed when the temperature was increased from 25 °C to 37 °C . PMID:25684450

  8. Use of 8-azidoguanosine 5'-(gamma-/sup 32/P)triphosphate as a probe of the guanosine 5'-triphosphate binding protein subunits in bovine rod outer segments

    SciTech Connect

    Kohnken, R.E.; Mc Connell, D.G.

    1985-07-02

    In an in vitro incubation, 8-azidoguanosine 5'-(gamma-/sup 32/P)triphosphate ( (gamma-/sup 32/P)-8-azido-GTP) labeled bleached rhodopsin independent of ultraviolet light. Characterization of this labeling indicated that rhodopsin was phosphorylated with (gamma-/sup 32/P)-8-azido-GTP as a phosphate donor. At low concentrations, ATP increased this labeling activity 5-fold. In the same incubation, (gamma-/sup 32/P)-8-azido-GTP also labeled G alpha (Mr 40 000). This labeling was ultraviolet light dependent. G beta (Mr 35 000) was also labeled dependent for the most part upon ultraviolet light, but a smaller component of labeling appeared to result from phosphorylation. Differential labeling of G alpha and G beta was found to vary intricately with experimental conditions, especially prebleaching of rhodopsin, tonicity of the medium, and the presence or absence of 2-mercaptoethanol. Affinity labeling of G alpha and G beta by (gamma-/sup 32/P)-8-azido-GTP in competition with ATP or GTP was kinetically complex, consistent with possible multiple binding sites for GTP on both subunits. Independent evidence for two or more binding sites on G alpha has been offered by other laboratories, and recently, at least one binding site on G beta and its analogues among the N proteins of adenylate cyclases has been identified.

  9. [32P]orthophosphate and [35S]methionine label separate pools of neurofilaments with markedly different axonal transport kinetics in mouse retinal ganglion cells in vivo.

    PubMed

    Nixon, R A; Lewis, S E; Mercken, M; Sihag, R K

    1994-11-01

    Newly synthesized neurofilament proteins become highly phosphorylated within axons. Within 2 days after intravitreously injecting normal adult mice with [32P]orthophosphate, we observed that neurofilaments along the entire length of optic axons were radiolabeled by a soluble 32P-carrier that was axonally transported faster than neurofilaments. 32P-incorporation into neurofilament proteins synthesized at the time of injection was comparatively low and minimally influenced the labeling pattern along axons. 32P-incorporation into axonal neurofilaments was considerably higher in the middle region of the optic axons. This characteristic non-uniform distribution of radiolabel remained nearly unchanged for at least 22 days. During this interval, less than 10% of the total 32P-labeled neurofilaments redistributed from the optic nerve to the optic tract. By contrast, newly synthesized neurofilaments were selectively pulse-labeled in ganglion cell bodies by intravitreous injection of [35S]methionine and about 60% of this pool translocated by slow axoplasmic transport to the optic tract during the same time interval. These findings indicate that the steady-state or resident pool of neurofilaments in axons is not identical to the newly synthesized neurofilament pool, the major portion of which moves at the slowest rate of axoplasmic transport. Taken together with earlier studies, these results support the idea that, depending in part on their phosphorylation state, transported neurofilaments can interact for short or very long periods with a stationary but dynamic neurofilament lattice in axons.

  10. Determination of 32P in urine for early estimation of the neutron exposure level for three victims of the JCO criticality accident.

    PubMed

    Nishimura, Yoshikazu; Takeda, Hiroshi; Miyamoto, Kiriko; Watanabe, Yoshito; Kouno, Fuyuki; Kuroda, Noriko; Kim, Hee Sun; Yukawa, Masae

    2002-03-01

    In the criticality accident which occurred on 30 September 1999 at a uranium conversion facility in Tokai-mura, Japan, three workers were severely exposed to neutron and gamma-ray irradiation. Preliminary estimations of doses from blood properties and 24Na concentration in blood were 16-20, 6-10 and 1-4 gamma-ray gray-equivalent (gammaGyEq) respectively for the three workers. For apparent dose estimation, neutron-induced radionuclides in biological materials such as blood, hair and urine were measured. Accordingly, we detected 32p in urine samples. The concentration ratios of 32P in the urine for the three workers showed a similar tendency to those of 24Na in blood. This result indicated that the radioactivity of 32P in urine could be used to estimate the neutron exposure level.

  11. Detection of bulky endogenous oxidative DNA lesions derived from 8,5'-cyclo-2'-deoxyadenosine by 32P-postlabeling assay

    PubMed Central

    Zhou, Guo-Dong; Moorthy, Bhagavatula

    2015-01-01

    8,5’-Cyclopurine-2’-deoxynucleotides represent a class of oxidative DNA lesions that are specifically repaired by nucleotide excision repair but not by base excision repair or direct enzymatic reversion. 32P-postlabeling assay is an ultrasensitive method that has been extensively used for the detection of carcinogen-DNA adducts in laboratory animal and epidemiological studies. This assay under modified chromatographic conditions is also a suitable and sensitive method for the detection of 8,5'-cyclo-2'-deoxyadenosine (cA). After enzymatic digestion of DNA, and enrichment of the oxidative products from the DNA digest, four dinucleotides containing cA, i.e. Ap-cAp, Cp-cAp, Gp-cAp, and Tp-cAp, are 5’-labeled with 32P-orthophosphate form [γ-32P]ATP mediated by polynucleotide kinase (PNK). The 32P-labeled cA products are separated by two-dimensional thin-layer chromatography (TLC) and quantified by Instant Imager or by a scintillation counter. The assay only requires 1–10 µg of DNA sample and is capable of detecting cA lesions as low as 1 in 1010 normal nucleotides. PMID:26344223

  12. Lethal modifications of DNA via the transmutation of 32P and 33P incorporated in the genome of the S13 bacteriophage.

    PubMed

    Cols, P; Apelgot, S; Guille, E

    1988-01-01

    When circular single-stranded DNA of phage S13 is labelled with 32P or 33P, the transmutations very efficiently bring about a loss of phage infectiousness (efficiency = 1 for 32P and 0.73 for 33P). For both radionuclides, the lethal efficiencies as well as the lethal events are different. In the case of 32P, the lethal event is the loss of the circular integrity of the DNA molecule, occurring as a consequence of a systematic single strand-break caused by each 32P decay (100%). Conversely, in the case of 33P, the lethal events are either a single strand-break (40%) or a local stereochemical modification (33%). The same primary event, the substitution at each 33P decay of a phosphate by a sulfate molecule, leads to one of these lethal events in relation to the decay site. Moreover, neither the phage adsorption nor its genome injection into bacteria depends on the physical state of the genome, and thus lethality is revealed at only the genetic level.

  13. Safety evaluation of 32P-chromic phosphate-poly L lactic acid particles interstitially implanted into livers of Beagle dogs.

    PubMed

    Liu, Lu; Huang, Peilin; Nie, Qi; Qi, Benzhong; Wu, Qinghua; Gao, Hailin; Yang, Zexuan; Chen, Daming

    2012-03-01

    The aim of this study was to investigate the safety and toxicity of biodegradable (32)P-chromic phosphate-poly L lactic acid ((32)P-CP-PLLA) particles interstitially implanted into Beagle dog livers. Eighteen healthy Beagle dogs were randomly divided into 6 groups (n=3), and were treated with drugs of different formulations or doses, as well as controls. At different time points after surgery, the experimental dogs were weighed. Detection of indicators of blood chemistry and liver fibrosis, SPECT bremsstrahlung imaging, computed tomography, histological examination, continuous blood measurement, and counting of urine and fecal radioactivity were performed for these dogs. SPECT imaging showed that after implantation of radioactive particles into livers, radioactivity continuously accumulated in the implanted sites, while no radioactivity imaging was found in the nonimplantation sites. The mean absorbance doses in the implantation sites were 89.8-178.7 Gy. Local spherical lesions were observed in tissues. The average effective half-life time of (32)P-CP-PLLA was 11.8 days. Within 4 weeks after surgery, slight or moderate swelling and degradation of liver cells were detected, while in 8 weeks after surgery, they are normal. For the blood chemistry, liver fibrosis, and other indicators, no significant differences were found between the control groups and particle implantation groups (F=1.378, p=0.232). (32)P-CP-PLLA particles have advantages including good targeting, immobile, being degradable in vivo, easy to be protected, and so on. It is suitable for treating solid tumors with blood supply. (32)P-CP-PLLA particles are a kind of safe, novel, radioactive implantation drug.

  14. Effect of iodide on glucose oxidation and /sup 32/P incorporation into phospholipids stimulated by different agents in dog thyroid slices

    SciTech Connect

    Tseng, F.Y.; Rani, C.S.; Field, J.B.

    1989-03-01

    Since iodide (I-) inhibits TSH stimulation of cAMP formation, which mediates most of the effects of the hormone, it has been assumed that this accounts for the inhibitory action of iodide on the thyroid. However, TSH stimulation of 32P incorporation into phospholipids and stimulation of thyroid metabolism by other agonists, such as carbachol, phorbol esters, and ionophore A23187, is not cAMP mediated. The present studies examined the effect of iodide on stimulation of glucose oxidation and 32P incorporation into phospholipids by TSH and other agonists to determine if the inhibition of cAMP formation was responsible for the action of iodide. Preincubation of dog thyroid slices for 1 h with iodide (10(-4) M) inhibited TSH-, (Bu)2cAMP-, carbachol-, methylene blue-, 12-O-tetradecanoyl phorbol-13-acetate-, ionophore A23187-, prostaglandin E1-, and cholera toxin-stimulated glucose oxidation. I- also inhibited the stimulation by TSH, 12-O-tetradecanoyl phorbol-13-acetate, carbachol, and ionophore A23187 of 32P incorporation into phospholipids. The inhibition was similar whether iodide was added 2 h before or simultaneously with the agonist. I- itself sometimes stimulated basal glucose oxidation, but had no effect on basal 32P incorporation into phospholipids. The effects of iodide on basal and agonist-stimulated thyroid metabolism were blocked by methimazole (10(-3) M). When dog thyroid slices were preloaded with 32PO4 or (1-14C)glucose, the iodide inhibition of agonist stimulation disappeared, suggesting that the effect of iodide involves the transport process. In conclusion, I- inhibited stimulation of glucose oxidation and 32P incorporation into phospholipids by all agonists, indicating that the effect is independent of the cAMP system and that iodide autoregulation does not only involve this system. Oxidation and organification of iodide are necessary for the inhibition.

  15. [The influence of CO2 and pH on (32)P-labelling of polyphosphates and organic phosphates in Ankistrodesmus braunii in the light].

    PubMed

    Ullrich, W R

    1971-03-01

    The effect of CO2 on the (32)P-labelling of polyphosphates and acid-soluble organic phosphates is studied in synchronously grown cultures of the green alga Ankistrodesmus braunii, using trichloroacetic acid treatment and acid hydrolysis for the fractionation of the phosphorus compounds.Three per cent CO2 in nitrogen causes an inhibition of the labelling of polyphosphates but a marked increase of (32)P in organic phosphates, whereas oxygen (CO2-free air) produces the reverse effect. Polyphosphates and ATP are the fractions most stimulated by O2, while stable organic phosphates show the strongest inhibition. Labelling of nucleic acids is relatively indifferent to both oxygen and CO2. Three per cent CO2 in air causes the same distribution of (32)P-labelling as 3 per cent CO2 in N2. (32)P-labelling is strongly dependent on the pH of the medium. In the absence of CO2, polyphosphate labelling is highest in the acidic range, whereas organic phosphates and ATP show optimum labelling and the highest percentage of the total (32)P in the alkaline pH range. The effect of CO2 is strongest between pH 5 and 6, that of oxygen between pH 8 and 9. Apparently the pH of the medium exerts a considerable influence upon the phosphate metabolism inside the cells.Increasing concentration of CO2 lead to the same change of (32)P-labelling in nitrogen as in air and to saturation at about 1 per cent CO2 under the conditions used. The curves are in good agreement with those of O2-evolution at increasing concentrations of CO2, but they show completely different rates.Young cells respond to CO2 and O2 differently from cells in the photosynthetically most active stage. In young cells both gasses are less effective.The effect of CO2 is explained by a strong increase in noncyclic photophosphorylation which can proceed only slowly in N2. ATP-consumption connected with high rates of CO2-fixation may be the reason for the low rates of (32)P-labelling in the polyphosphate fraction when CO2 is present

  16. CARS spectroscopy of the NaH2 collision complex: the nature of the Na(32 P)H2 exciplex — ab initio calculations and experimental results

    NASA Astrophysics Data System (ADS)

    de Vivie-Riedle, R.; Hering, P.; Kompa, K. L.

    1990-12-01

    CARS (Coherent Anti-Stokes Raman Scattering) has been used to analyze the rovibronic state distribution of H2 after collision with Na(32 P). New lines, which do not correspond to H2 lines are observed in the CARS spectrum. The experiments point to the formation of a complex of Na(32 P)H2 in A 2 B 2 symmetry. Ab initio calculations of the A 2 B 2 potential were performed. On this surface the vibrational spectra of the exciplex are evaluated. The observed lines can be attributed to vibrational transitions in the complex, in which combinational modes are involved. The connection of experimental and theoretical results indicates that a collisionally stabilized exciplex molecule is formed during the quenching process.

  17. Comparative analysis of aromatic DNA adducts in fish from polluted and unpolluted areas by the sup 32 P-postlabeling analysis

    SciTech Connect

    Tsungyun Liu; Shuling Cheng; Chinwen Chi ); Tzuuhuei Ueng; Yunefang Ueng )

    1991-11-01

    The {sup 32}P-postlabeling technique, developed by Randerath and his colleagues, is a highly sensitive assay to detect the damaged DNA as DNA-carcinogen adduct. Consequently, measurement of this DNA carcinogen adduct concentrations in target tissues of organisms may provide a key biologic end-point of exposure to environmental carcinogens. In this study the authors analyze the hepatic DNA from a bottom-feeding fish, tilapia (Tilapia mossambica), sampled from the down stream Damsui River where fish are exposed to high levels of polycyclic aromatic hydrocarbons (PAH) in sediment and from the upstream Fe-Tsui reservoir where the water has been used as the source of drinking water for Taipei Metropolitan area. In doing so, they will be able to determine whether this {sup 32}P-postlabeling analysis of hepatic DNA from the bottom-feeding tilapia can be used to monitor large molecule aromatic carcinogens in the environment.

  18. [Uptake and phosphorylation of exogenous substrates in Ankistrodesmus braunii : II. Effect of glycolic acid upon the incorporation of (32)P-labelled phosphate in dark and in light].

    PubMed

    Lysek, G; Simonis, W

    1968-12-01

    The findings of JACOBI (1959) that glycolate enhances the uptake and the incorporation of (32)P-labelled orthophosphate were re-investigated. In contrast to the findings of JACOBI we found that glycolic acid has no effect at concentrations between 10(-3) and 10(-7)M. The effect reported by JACOBI was seen only when instead of glycolic acid Na-glycolate was used in the experiments, as it was done by JACOBI. Further experiments showed that the enhancement of (32)P-incorporation is due only to the Na(+)-ions and not to glycolic acid. In addition it was found that up to pH 4,3 no glycolic acid is resorbed from the medium by cells of Ankistrodesmus braunii.

  19. Determination of the ATP Affinity of the Sarcoplasmic Reticulum Ca(2+)-ATPase by Competitive Inhibition of [γ-(32)P]TNP-8N3-ATP Photolabeling.

    PubMed

    Clausen, Johannes D; McIntosh, David B; Woolley, David G; Andersen, Jens Peter

    2016-01-01

    The photoactivation of aryl azides is commonly employed as a means to covalently attach cross-linking and labeling reagents to proteins, facilitated by the high reactivity of the resultant aryl nitrenes with amino groups present in the protein side chains. We have developed a simple and reliable assay for the determination of the ATP binding affinity of native or recombinant sarcoplasmic reticulum Ca(2+)-ATPase, taking advantage of the specific photolabeling of Lys(492) in the Ca(2+)-ATPase by [γ-(32)P]2',3'-O-(2,4,6-trinitrophenyl)-8-azido-adenosine 5'-triphosphate ([γ-(32)P]TNP-8N3-ATP) and the competitive inhibition by ATP of the photolabeling reaction. The method allows determination of the ATP affinity of Ca(2+)-ATPase mutants expressed in mammalian cell culture in amounts too minute for conventional equilibrium binding studies. Here, we describe the synthesis and purification of the [γ-(32)P]TNP-8N3-ATP photolabel, as well as its application in ATP affinity measurements.

  20. 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.

  1. Evaluation of the biotinylated (Blugene) vs sup 32 P-labeled cDNA probes of beta-glucocerebrosidase: Relative sensitivities in genomic and other systems

    SciTech Connect

    Strasberg, P. )

    1989-07-01

    The sensitivity, rapidity, and ease of use of biotinylated (Blugene) and {sup 32}P cDNA probes have been compared, the probe being the cDNA for beta-glucocerebrosidase. With the Blugene kit I could detect 2 pg of biotinylated DNA on dot blots. However, under conditions of hybridization, the lower limit of detection for unlabeled cDNA (transblotted onto nitrocellulose) by its labeled counterpart was 5000-fold smaller (10 pg vs 50 ng) for the isotopically labeled probe. {sup 32}P- and Blugene-probes hybridized detectably with 0.5 and 10 micrograms, respectively, of transblotted EcoR 1-digested genomic DNA, making the radioactive method 20 times as sensitive. However, color development was complete within 30 min to 3 h, whereas radioautoradiography required 12 h to one week. Blugene was also safer, easy to use, and effective under appropriate conditions. The {sup 32}P method is expensive, hazardous, time-consuming, and technically difficult. This nonisotopic procedure represents a desirable improvement in biotechnology.

  2. Development of a 32P-postlabelling method for the analysis of 2'-deoxyguanosine-3'-monophosphate and DNA adducts of methylglyoxal.

    PubMed

    Vaca, C E; Fang, J L; Conradi, M; Hou, S M

    1994-09-01

    A 32P-postlabelling assay was developed for the analysis of adducts arising from the reaction of 2'-deoxyguanosine-3'-monophosphate with the 1,2-dicarbonyl compound methylglyoxal, a major mutagen in several foodstuffs, in particular, instant and brewed coffee. The 32P-postlabelling reaction was optimized by testing various parameters such as the kinetics of phosphorylation by T4 polynucleotide kinase, substrate concentration-dependent labelling efficiency and the concentration of the various ingredients of the phosphorylation reaction. The sensitivity to the 3'-monophosphate dephosphorylation activity of nuclease P1 was also studied. Four isomeric reaction products were separated by HPLC, structurally characterized and identified as 3-(2'-deoxy-beta-D-erythro-pentafuranosyl)-6,7-dihydro-6,7-dihydro xy-6- methylimidazo[2,3-b]purine-9(8H)one. The same adducts could be detected from calf thymus DNA that had been reacted in vitro with methylglyoxal. DNA adducts were isolated after enzymatic digestion to mononucleotides followed by nuclease P1 digestion of normal nucleotides. The total level of methylglyoxal-DNA adducts obtained was 5.7 +/- 1.7 (n = 15) adducts/10(6) nucleotides. The 32P-postlabelling method was further validated by the detection of adducts of methylglyoxal in DNA from freshly isolated and stimulated human lymphocytes exposed in vitro. The concentrations of the adducts detected in these samples were 8.2 +/- 0.9 (n = 3) adducts/10(7) nucleotides and 1.5 +/- 0.1 (n = 3) adducts/10(6) nucleotides after treatment with 1.5 and 3.0 mM methylglyoxal respectively.

  3. /sup 32/P-postlabeling analysis of DNA adducts in liver of wild English sole (Parophrys vetulus) and winter flounder (Pseudopleuronectes americanus)

    SciTech Connect

    Varanasi, U.; Reichert, W.L.; Stein, J.E.

    1989-03-01

    The 1-butanol adduct enhancement version of the 32P-postlabeling assay was used to measure the levels of hepatic DNA adducts in the marine flatfish, English sole (Parophrys vetulus), sampled from the Duwamish Waterway and Eagle Harbor, Puget Sound, WA, where they are exposed to high concentrations of sediment-associated chemical contaminants and exhibit an elevated prevalence of hepatic neoplasms. Hepatic DNA was also analyzed from English sole from a reference area (Useless Bay, WA) and from reference English sole treated with organic-solvent extracts of sediments from the two contaminated sites. Autoradiograms of thin-layer chromatograms of 32P-labeled hepatic DNA digests from English sole from the contaminated sites exhibited up to three diagonal radioactive zones, which were not present in autoradiograms of thin-layer chromatogram maps of 32P-labeled DNA digests from English sole from the reference site. These diagonal radioactive zones contained several distinct spots as well as what appeared to be multiple overlapping adduct spots. The levels (nmol of adducts/mol of nucleotides) of total DNA adducts for English sole from Duwamish Waterway and Eagle Harbor were 26 +/- 28 (DS) and 17 +/- 9.6, respectively. All autoradiograms of DNA from fish from the contaminated sites exhibited a diagonal radioactive zone where DNA adducts of chrysene, benzo(a)pyrene, and dibenz(a,h)anthracene, formed in vitro using English sole hepatic microsomes, were shown to chromatograph. English sole treated with extracts of the contaminated sediments had adduct profiles generally similar to those for English sole from the respective contaminated sites.

  4. Detection of calmodulin-binding proteins using a 32P-labeled GST-calmodulin fusion protein and a novel renaturation protocol.

    PubMed

    Fischer, R; Wei, Y; Berchtold, M

    1996-08-01

    To identify calmodulin-binding proteins in cellular extracts and tissue homogenates and to analyze purified calmodulin target proteins, overlay procedures using 125I-calmodulin or, more recently, nonradioactive biotinylated calmodulin have been widely used. Here we describe a rapid, alternative method for detecting calmodulin-binding proteins with a 32P-labeled calmodulin probe generated as a glutathione-S-transferase (GST)-fusion protein. We used a modified pGEX-2TK vector, which contains the flag epitope and the consensus sequence R-R-A-S, that can be phosphorylated by the cAMP-dependent protein kinase A. The fusion protein is easily purified from bacterial bysates by affinity chromatography using glutathione-Sepharose 4B beads. Phosphorylation of GST-calmodulin is performed directly on the beads and, after elution with reduced glutathione, the labeled calmodulin probe can be used for overlay experiments. We also describe a rapid renaturation protocol that enhances the signal for some but not all calmodulin-binding proteins and is used after the proteins have been transferred to nitrocellulose filters. Furthermore, we have compared the specificity and sensitivity of the 32P-labeled GST-calmodulin overlay with those of 125I-calmodulin and biotinylated calmodulin, clearly indicating that our newly developed protocol is a suitable alternative to conventionally used calmodulin overlay procedures.

  5. Detection of the acrolein-derived cyclic DNA adduct by a quantitative 32P-postlabeling/solid-phase extraction/HPLC method: blocking its artifact formation with glutathione.

    PubMed

    Emami, Armaghan; Dyba, Marcin; Cheema, Amrita K; Pan, Jishen; Nath, Raghu G; Chung, Fung-Lung

    2008-03-01

    Acrolein (Acr), a hazardous air pollutant, reacts readily with deoxyguanosine (dG) in DNA to produce cyclic 1, N2-propanodeoxyguanosine adducts (Acr-dG). Studies demonstrate that these adducts are detected in vivo and may play a role in mutagenesis and carcinogenesis. In the study described here, a quantitative 32P-postlabeling/solid-phase extraction/HPLC method was developed by optimizing the solid-phase extraction and the 32P-postlabeling conditions for analysis of Acr-dG in DNA samples with a detection limit of 0.1 fmol. It was found that Acr-dG can form as an artifact during the assay. Evidence obtained from mass spectrometry indicates that the Acr in water used in the assay is a likely source of artifact formation of Acr-dG. The formation of Acr-dG as an artifact can be effectively blocked by adding glutathione (GSH) to the DNA sample to be analyzed. In addition, Acr-dG was detected as a contaminant in the commercial dG and dT 3'-monophosphate samples. Finally, this method was used to detect Acr-dG in calf thymus and human colon HT29 cell DNA with an excellent linear quantitative relationship.

  6. Uptake and distribution of sup 203 Hg by fish fingerlings, Cirrhina mrigala, exposed to linear alkyl benzene sulfonate

    SciTech Connect

    Misra, V.; Kumar, V.; Pandey, S.D.; Viswanathan, P.N. )

    1989-07-01

    Ecological changes caused by the continued pollution of the aquatic environment by chemicals through industrial effluents and domestic sewage and emanations settling into water pose grave concern. Synthetic detergents are one of the most important in this respect since they find their way into aquatic ecosystems thereby affecting the food chain. Earlier studies with diverse aquatic fauna and flora suggested the potential ecotoxicological impact of synthetic detergents. A large number of reports are available on the pollutants toxic to fish. Fish are known to accumulate mercury by virtue of efficient uptake and slow rate of elimination. Mercury also causes morphological and physiological defects with consequent behavioral abnormalities in fish. Even though in actual situations the stress to the ecosystem is caused by a mixture of pollutants, the interactive effect of two or more pollutants present together is poorly understood. Also, in the presence of one toxicant, the capacity of the ecosystem to deal with others can be impaired so that even biodegradable water pollutants may tend to accumulate. Therefore, an attempt has been made to study the uptake and distribution of mercury in presence and absence of detergent to test for any combined effects.

  7. Behavior of mercury in bio-systems. II. Depuration of /sup 203/Hg/sup 2 +/ in various trophic levels

    SciTech Connect

    Hamdy, M.K.; Prabhu, N.V.

    1984-01-01

    Using radiotracer techniques, the depuration rates for methylmercury at three trophic levels in an aquatic ecosystem are examined. Bacteria (decomposers), mosquito larvae (primary consumers), and fish (secondary consumers) were studied. Results indicated that depuration rates for mercury were temperature dependent - the rate of depuration increased with increase in temperature (up to 45/sup 0/C)

  8. 32P-postlabeling analysis of adducts formed between DNA and safrole 2',3'-epoxide: absence of adduct formation in vivo.

    PubMed

    Qato, M K; Guenthner, T M

    1995-01-01

    We have used the 32P-postlabeling technique to examine the binding of safrole 2',3'-oxide to DNA. At least 8 covalent adducts are formed when calf thymus DNA is incubated with this oxygenated metabolite of safrole in vitro. However, no corresponding adducts are formed with liver DNA when whole animals are exposed to safrole 2',3'-oxide, or safrole itself. Although safrole 2',3'-oxide is readily formed in vivo, and is sufficiently reactive to covalently bind to DNA, it is probably not a factor in the in vivo genotoxicity of safrole. We also demonstrate that adducts with similar mobility to the major safrole 2',3'-oxide-DNA adduct are formed in vitro between safrole 2',3'-oxide and deoxyguanosine, and also between its chemical analogs allylbenzene 2',3'-oxide or estragole 2',3'-oxide and DNA.

  9. sup 32 P-postlabeling detection of radiation-induced DNA damage: Identification and estimation of thymine glycols and phosphoglycolate termini

    SciTech Connect

    Weinfeld, M.; Soderlind, K.J.M. )

    1991-01-29

    A {sup 32}P-postlabeling assay has been developed that permits detection of several radiogenic base and sugar lesions of DNA at the femtomole level. The technique is based on the inability of DNase I and snake venom phosphodiesterase to cleave the internucleotide phosphodiester bond immediately 5{prime} to the site of damage so that complete digestion of irradiated DNA with these nucleases and alkaline phosphatase yields lesion-bearing dinucleoside monophosphates. Because these fragments contain an unmodified nucleoside at the 5{prime}-end of each molecule, they can be readily phosphorylated by T4 polynucleotide kinase and ({gamma}-{sup 32}P)ATP and analyzed by polyacrylamide gel electrophoresis and reverse-phase HPLC. The authors observed a linear induction of total damage in DNA irradiated with 5-50 Gy. Virtually no damage was detected when the DNA was irradiated in solution containing 1 M DMSO, implicating hydroxyl radicals in the formation of these lesions. Evidence for the presence of thymine glycols and phosphoglycolate groups came from (1) a comparison of the radiation-induced products with those produced by OsO{sub 4} and KMnO{sub 4} and (2) incubation of irradiated DNA with Escherichia coli endonuclease 3 and exonuclease 3 before analysis by the postlabeling procedure. This confirmed by comigration of the radiogenic products with chemically synthesized markers. The identity of the 5{prime}-nucleotide of each isolated compound was obtained by nuclease P1 digestion. This analysis of nearest-neighbor bases to thymine glycols and phosphoglycolates indicated a nonrandom interaction between radiation-induced hydroxyl radicals and DNA.

  10. 32P-post-labelling of 7-(3-chloro-2-hydroxypropyl)guanine in white blood cells of workers occupationally exposed to epichlorohydrin.

    PubMed

    Plna, K; Osterman-Golkar, S; Nogradi, E; Segerbäck, D

    2000-02-01

    Epichlorohydrin (ECH) is a simple 3-carbon epoxide of industrial importance. It has been shown to be genotoxic in several systems and carcinogenic in experimental animals. The aim of the present investigation was to study DNA adducts of ECH as a biomarker of occupational exposure to this chemical. 7-(3-Chloro-2-hydroxypropyl)guanine (7-CHP-guanine) was analysed in DNA from white blood cells using an anion exchange-based adduct enrichment protocol of the (32)P-post-labelling/HPLC-based assay. Blood samples were collected from seven workers handling ECH (exposed), nine workers not handling ECH but normally present in the premises where this chemical is used (potentially exposed) and 13 office and factory workers from locations in the plant where ECH is not handled (controls). 7-CHP-guanine was detected in five of the seven workers exposed to ECH (1.6-7.1 mol/10(9) mol nucleotides) and in two of the nine workers potentially exposed to ECH (0.8-1.5 mol/10(9) mol nucleotides). This adduct was not detected in any of the 13 controls. The difference in adduct levels between exposed workers and controls was statistically significant (Mann-Whitney test, P < 0.001), as was the difference between exposed workers and potentially exposed workers (P = 0.017). The recovery of 7-CHP-guanine in the (32)P-post-labelling assay was on average 48 +/- 7%, which is considerably higher than previously reported for other 7-alkylguanines. The method used had a limit of detection of approximately 0.4 mol adduct/10(9) mol nucleotides using 20 microg DNA. This study shows for the first time ECH-induced DNA adducts in humans and suggests that 7-CHP-guanine may be used as a biomarker of occupational exposure to ECH.

  11. Improvement of Arbuscular Mycorrhiza Development by Inoculation of Soil with Phosphate-Solubilizing Rhizobacteria To Improve Rock Phosphate Bioavailability ((sup32)P) and Nutrient Cycling

    PubMed Central

    Toro, M.; Azcon, R.; Barea, J.

    1997-01-01

    The interactive effect of phosphate-solubilizing bacteria and arbuscular mycorrhizal (AM) fungi on plant use of soil P sources of low bioavailability (endogenous or added as rock phosphate [RP] material) was evaluated by using soil microcosms which integrated (sup32)P isotopic dilution techniques. The microbial inocula consisted of the AM fungus Glomus intraradices and two phosphate-solubilizing rhizobacterial isolates: Enterobacter sp. and Bacillus subtilis. These rhizobacteria behaved as "mycorrhiza helper bacteria" promoting establishment of both the indigenous and the introduced AM endophytes despite a gradual decrease in bacterial population size, which dropped from 10(sup7) at planting to 10(sup3) CFU g(sup-1) of dry rhizosphere soil at harvest. Dual inoculation with G. intraradices and B. subtilis significantly increased biomass and N and P accumulation in plant tissues. Regardless of the rhizobacterium strain and of the addition of RP, AM plants displayed lower specific activity ((sup32)P/(sup31)P) than their comparable controls, suggesting that the plants used P sources not available in their absence. The inoculated rhizobacteria may have released phosphate ions ((sup31)P), either from the added RP or from the less-available indigenous P sources, which were effectively taken up by the external AM mycelium. Soluble Ca deficiency in the test soil may have benefited P solubilization. At least 75% of the P in dually inoculated plants derived from the added RP. It appears that these mycorrhizosphere interactions between bacterial and fungal plant associates contributed to the biogeochemical P cycling, thus promoting a sustainable nutrient supply to plants. PMID:16535730

  12. Post-stenting Intravascular Brachytherapy Trials on Hypercholesterolemic Rabbits Using 32P Liquid Sources: Implications for Prevention of In-Stent Restenosis

    SciTech Connect

    Wilczek, Krzysztof; Walichiewicz, Piotr; Petelenz, Barbara; Jachec, Wojciech; Jochem, Jerzy; Tomasik, Andrzej; Bilski, Pawel; Snietura, Miroslaw; Wodniecki, Jan

    2002-08-15

    Purpose: Liquid sources of radiation delivered in angioplasty balloons may be a convenient self-centering device used for prevention of in-stent restenosis. To test the effectiveness of this method an intravascular brachytherapy study was performed using 32P liquid sources in an animal model. Methods: The radial dose distribution around angioplasty balloons filled with solutions of Na2H32PO4 was calibrated by thermoluminescence dosimetry. The animal experiments were performed in rabbits with induced hypercholesterolemia. The balloons containing 32P were introduced into iliac arteries immediately after stent implantation. Estimated 7-49 Gy doses required 30-100 minirradiations. Radiation effects were evaluated by comparing the thickness of various components of the artery wall. Results:Doses of 7, 12, 16 or 49 Gy on the internal artery surface required 30-100 min of irradiation. The dose of 49 Gy at 'zero' distance corresponding to 16 Gy at 1.0 mm from the balloon surface reduced hypertrophy in every layer of the arterial wall: in the intima the cross-sectional areas were 0.13 versus 0.91 mm2, in the media were 0.5 versus 0.46 mm2 and in the adventitia were 0.04 versus 0.3 mm2 (p <0.05). A dose of 7 Gyat the balloon surface produced adverse irradiation effects: the intimal area of the artery was 2.087 versus 0.857 mm2, the medial area was 0.59 versus 0.282 mm2 and the adventitial area was 0.033 versus 0.209 mm2 in treated and control arteries, respectively.Conclusion: Application of a 49 Gy irradiation dose to the internal arterial surface effectively prevented in-stentrestenosis.

  13. Examination of microsomal cytochrome P450-catalyzed in vitro activation of o-phenylphenol to DNA binding metabolite(s) by 32P-postlabeling technique.

    PubMed

    Pathak, D N; Roy, D

    1992-09-01

    It has been previously reported that the reactive metabolites phenylsemiquinone and phenylbenzoquinone are generated during microsomal cytochrome P450-catalyzed redox cycling of o-phenylphenol (OPP). However, covalent modification of DNA by OPP-reactive metabolites has yet not been demonstrated. In the present study we have investigated the covalent binding in DNA by OPP-reactive metabolites using 32P-postlabeling. Analysis of adducts by 32P-postlabeling in products of chemical reaction of DNA with phenylbenzoquinone revealed four major and several minor adducts. The chemical reaction of deoxyguanosine 3'-phosphate with phenylbenzoquinone also showed four major adducts. The chromatographic mobility of major adducts of deoxyguanosine 3'-phosphate-phenylbenzoquinone was identical to that of major adducts of DNA-phenylbenzoquinone. The major adducts are demonstrated to be stable. The total covalent binding in deoxyguanosine 3'-phosphate by phenylbenzoquinone (686,000-687,000 amol/nmol nucleotide) was higher than that observed in DNA (26,500-28,000 amol/nmol nucleotides). Reaction of DNA with OPP or a hydroxylated metabolite of OPP, phenylhydroquinone, in the presence of microsomes and NADPH or cumene hydroperoxide showed four major adducts. Adduct formation in DNA by OPP or phenylhydroquinone in the presence of the microsomal activation system was drastically decreased by known inhibitors of cytochrome P450. The chromatographic mobility of major adducts in DNA by OPP or phenylhydroquinone in the presence of microsomal activation system matched with those major adducts observed in deoxyguanosine 3'-phosphate or DNA reacted with pure phenylbenzoquinone. These data demonstrate that OPP or phenylhydroquinone, a hydroxylated metabolite of OPP, is able to bind covalently to DNA in the presence of a microsomal cytochrome P450 activation system. Phenylbenzoquinone is one of the DNA-binding metabolite(s) of OPP. It is concluded that OPP is genotoxic in an in vitro system and

  14. A novel approach to brachytherapy in hepatocellular carcinoma using a phosphorous{sup 32} ({sup 32}P) brachytherapy delivery device-a first-in-man study

    SciTech Connect

    Goh, Anthony Soon-Whatt; Chung, Alexander Yaw-Fui; Lo, Richard Houa-Gong; Lau, T.-N.; Yu, Sidney Wing-Kwong; Chng, May; Satchithanantham, Somanesan; Loong, Susan Li-Er; Ng, David Chee-Eng; Lim, Beng-Choo; Connor, Stephen; Chow, Pierce Kah-Hoe . E-mail: gsupc@singnet.com.sg

    2007-03-01

    Purpose: While potentially very useful, percutaneously delivered brachytherapy of inoperable intra-abdominal solid tumors faces significant technical challenges. This first-in-man study is designed to determine the safety profile and therapeutic efficacy of a novel phosphorous ({sup 32}P) brachytherapy device (BrachySil) in patients with unresectable hepatocellular carcinoma. Methods and Materials: Patients received single percutaneous and transperitoneal implantations of BrachySil under local anesthesia directly into liver tumors under ultrasound or computed tomographic guidance, at an activity level of 4 MBq/cc of tumor. Toxicity was assessed by the nature, incidence, and severity of adverse events (Common Toxicity Criteria scores) and by hematology and clinical chemistry parameters. Target tumor response was assessed with computed tomographic scans at 12 and 24 weeks postimplantation using World Health Organization criteria. Results: Implantations were successfully carried out in 8 patients (13-74 MBq, mean 40 MBq per tumor) awake and under local anesthesia. Six of the 8 patients reported 19 adverse events, but no serious events were attributable to the study device. Changes in hematology and clinical chemistry were similarly minimal and reflected progressive underlying hepatic disease. All targeted tumors were responding at 12 weeks, with complete response (100% regression) in three lesions. At the end of the study, there were two complete responses, two partial responses, three stable diseases, and one progressive disease. Conclusion: Percutaneous implantation of this novel {sup 32}P brachytherapy device into hepatocellular carcinoma is safe and well tolerated. A significant degree of antitumor efficacy was demonstrated at this low dose that warrants further investigation.

  15. [The effect of oxygen on the (32)P-labelling of polyphosphates and organic phosphates in Ankistrodesmus braunii in the light].

    PubMed

    Ullrich, W R

    1970-09-01

    Short time incorporation of (32)P was carried out with synchronised algae (young cells) depleted of phosphate. For the separation and determination of the acid-insoluble phosphate fractions of the cells an improved fractionation procedure was applied. In order to exclude competition by carbon dioxide all experiments were done in the absence of CO2.Compared with nitrogen, CO2-free air produces an increase in the labelling of phosphorylated compounds in the light. In strong white light, at high pH, air effects a remarkable increase of (32)P in the acid-insoluble phosphate (P u), mainly in inorganic polyphosphates (P ul), whereas the total phosphate uptake remains almost unchanged. The increase in labelling of acid-insoluble phosphate is, therefore, accompanied by a substantial decrease in the labelling of acid-soluble compounds (P l). In weak white light or in far-red light, at low pH even in strong white or red light, an increase of phosphate uptake and an increased labelling of the acid-stable organic acid-soluble fraction (P os) is observed instead. The effect of oxygen increases somewhat with increasing light intensity up to light saturation, and it increases markedly with increasing oxygen concentration.An essential contribution by oxidative phosphorylation to this oxygen effect can be ruled out on account of its much higher sensitivity to oxygen. Pseudocyclic photophosphorylation is also not regarded as the main force because of its higher oxygen affinity. Occurrence of photorespiration has not been clearly established so far in related algae (Chlorella), and its use for phosphorylation is unknown. A better, although not complete explanation is given by comparing the oxygen effect with the well-known inhibition of photosynthesis by oxygen (Warburg effect), which leads to an increase in glycolate formation and a simultaneous decrease in the pool sizes of carbon reduction cycle intermediates, even in the absence of CO2. Since the photophosphorylation process, as

  16. 32P-post-labelling analysis of DNA adducts formed in the upper gastrointestinal tissue of mice fed bracken extract or bracken spores.

    PubMed Central

    Povey, A. C.; Potter, D.; O'Connor, P. J.

    1996-01-01

    Bracken toxicity to both domestic and laboratory animals is well established and tumours are formed when rodents are treated with either bracken extracts or bracken spores. In this study we have administered bracken spores and extract to mice in order to investigate whether such exposure leads to the formation of DNA adducts. DNA, isolated from the upper gastrointestinal tract and liver, was digested to 3'-nucleotides. Adducts were extracted with butanol, 32P-post-labelled, separated by thin layer chromatography (TLC) and visualised and quantified using storage-phosphor technology. A cluster of adducts was clearly seen in the DNA of the upper gastrointestinal tract, but not liver, 5 and 24 h after treatment with bracken extract or bracken spores. These adducts were not observed in DNA extracted from vehicle-treated animals. Whereas, after 5 h adduct levels in extract and spore-treated animals were similar, after 24 h adduct levels in the extract-treated animals had diminished by > 75%, but levels in spore-treated animals remained similar to those found after 5 h. This suggests that the DNA-reactive compounds were being released slowly from the spores, even though the spores had been sonicated before administration. Adducts were also quantified after the addition of an internal standard (deoxyinosine 3'-monophosphate) by comparing the amount of label incorporated into the adducts with that found in a known amount of the internal standard. Adduct levels using this internal standard approach were similar to those found by direct measurement of radioactivity incorporated into the adduct, indicating that the labelling of adducts was quantitative. We have tried, unsuccessfully, to synthesise ptaquiloside, the principal carcinogenic component present within bracken. However, similar patterns of adducts were observed when two other compounds, (1-(4-chlorophenyl sulphonyl)-l-cyclopropane carbonitrile and 3-cyclopropylindeno [1,2-c] pyrazol-4-(O-methyl)oxime), which both

  17. The structure of the human intron-containing S8 ribosomal protein gene and determination of its chromosomal location at 1p32-p32. 4

    SciTech Connect

    Davies, B.; Fried, M. )

    1993-01-01

    The intron-containing gene encoding human ribosomal protein SS (RPS8) has been cloned and characterized, and its chromosomal position determined. Using a PCR-based cloning strategy, we have isolated the intron-containing gene in the presence of its many processed pseudogenes and determined the DNA sequence of the entire gene and its upstream and downstream flanking regions. The human RPS8 gene is 3161 bp in length and comprises six exons. Despite lacking a consensus TATA box, primer extension analysis indicates that the start of transcription is precisely located at a C residue within an 11-bp oligopyrimidine tract. The first exon, which contains the ATG start codon, is just 27 bp in length. The DNA sequence 5[prime] to the RPS8 gene and within the first exon and intron shows several features of a CpG island. A combination of Southern blotting, PCR, and fluorescence in situ hybridization analyses has enabled the chromosomal location of the human RPSS gene to be determined as lp32-p34.1. 51 refs., 5 figs.

  18. Stereoselective release of polycyclic aromatic hydrocarbon-deoxyadenosine adducts from DNA by the 32P postlabeling and deoxyribonuclease I/snake venom phosphodiesterase digestion methods

    SciTech Connect

    Cheh, A.M.; Yagi, H.; Jerina, D.M. )

    1990-11-01

    The restricted ability of deoxyribonuclease I/snake venom phosphodiesterase digestion to liberate deoxyadenosine (dA) nucleotide adducts of polycyclic aromatic hydrocarbons from DNA, first observed by Dipple and Pigott with the bay-region diol epoxide adducts of 7,12-dimethylbenz(a)anthracene, has been observed with the dA adducts of benz(a)anthracene and benzo(c)phenanthrene diol epoxides. The micrococcal nuclease/spleen phosphodiesterase digestion used in the original 32P postlabeling procedure developed by Randerath to determine DNA adducts also failed to liberate dA nucleotide adducts quantitatively. Thus either method can potentially lead to an underestimation of the extent to which dA has been modified in DNA. The two digestion procedures exhibit systematic and mostly opposite stereoselectivity in the pattern of which dA adducts are resistant to digestion, which suggest that these adducts may have preferred orientations within modified DNA that are determined by whether they have the R or S configuration at C-1, the point of attachment between the exocyclic amino group of dA and the hydrocarbon; this in turn is dictated by the configuration about the precursor benzylic epoxide carbon and the cis versus trans nature of epoxide opening during adduct formation.

  19. 32P-postlabeling DNA adduct assay: cigarette smoke-induced dna adducts in the respiratory and nonrespiratory rat tissues. Book chapter

    SciTech Connect

    Gupta, R.C.; Gairola, C.G.

    1990-01-01

    An analysis of the tissue DNA adducts in rats by the sensitive (32)p-postlabeling assay showed one to eight detectable DNA adducts in lung, trachea, larynx, heart and bladder of the sham controls. Chronic exposure of animals to mainstream cigarette smoke showed a remarkable enhancement of most adducts in the lung and heart DNA. Since cigarette smoke contains several thousand chemicals and a few dozen of them are known or potential carcinogens, the difference between the DNA adducts of nasal and the other tissues may reflect the diversity of reactive constituents and their differential absorption in different tissues. In comparison to the lung DNA adducts, the adducts in nasal DNA were less hydrophobic. Identity of the predominant adducts was further investigated by comparison with several reference DNA adducts from 10 PAH and aromatic amines. Since some of these chemicals are present in cigarette smoke, the results suggest that these constituents of cigarette smoke may not be directly responsible for formation of DNA adducts in the lung and heart of the smoke-exposed animals.

  20. Assignment of the muscle-eye-brain disease gene to 1p32-p34 by linkage analysis and homozygosity mapping.

    PubMed Central

    Cormand, B; Avela, K; Pihko, H; Santavuori, P; Talim, B; Topaloglu, H; de la Chapelle, A; Lehesjoki, A E

    1999-01-01

    Muscle-eye-brain disease (MEB) is an autosomal recessive disease of unknown etiology characterized by severe mental retardation, ocular abnormalities, congenital muscular dystrophy, and a polymicrogyria-pachygyria-type neuronal migration disorder of the brain. A similar combination of muscle and brain involvement is also seen in Walker-Warburg syndrome (WWS) and Fukuyama congenital muscular dystrophy (FCMD). Whereas the gene underlying FCMD has been mapped and cloned, the genetic location of the WWS gene is still unknown. Here we report the assignment of the MEB gene to chromosome 1p32-p34 by linkage analysis and homozygosity mapping in eight families with 12 affected individuals. After a genomewide search for linkage in four affected sib pairs had pinpointed the assignment to 1p, the MEB locus was more precisely assigned to a 9-cM interval flanked by markers D1S200 proximally and D1S211 distally. Multipoint linkage analysis gave a maximum LOD score of 6.17 at locus D1S2677. These findings provide a starting point for the positional cloning of the disease gene, which may play an important role in muscle function and brain development. It also provides an opportunity to test other congenital muscular dystrophy phenotypes, in particular WWS, for linkage to the same locus. PMID:9915951

  1. Sonographic genital ambiguity in a fetus due to a mosaic 45,X/46,X,idic(Y)(qter-p11.32::p11.32-qter) karyotype.

    PubMed

    Marcus-Soekarman, D; Hamers, G; Mulder, A L M; Offermans, Jeanne; Offermans, Jos; Engelen, J; van Lent-Albrechts, J C M; Robben, S G F; de Muinck Keizer-Schrama, S; Wolffenbuttel, K P; Looijenga, L H J; Oosterhuis, J W; Schrander-Stumpel, C; Nijhuis, J

    2005-04-01

    Nowadays, improved ultrasound techniques enable the detection of more subtle congenital abnormalities at an earlier stage of fetal development. Current cytogenetic techniques can characterize a chromosomal abnormality in greater detail. These advancements in both diagnostic possibilities have helped to answer many questions but have also created new issues and dilemmas in counselling. This is illustrated by this case report of a 35-year-old woman, who presented at the end of the second trimester of her first pregnancy. Sonographic examination indicated an abnormal external genital in a male fetus. A differential diagnosis of hypospadia was made. During follow-up, an amniocentesis was performed, and this showed a 45,X/46,X,idic(Y)(qter-p11.32::p11.32-qter) karyotype as the cause of the sonographic findings. Cytogenetic characterization of the isodicentric Y chromosome and pre- and post-natal findings in the child are reported. Cases with a similar karyotype reported in the literature are reviewed.

  2. {sup 32}P Brachytherapy Conformal Source Model RIC-100 for High-Dose-Rate Treatment of Superficial Disease: Monte Carlo Calculations, Diode Measurements, and Clinical Implementation

    SciTech Connect

    Cohen, Gil'ad N.; Munro, John J.; Kirov, Assen; Losasso, Thomas; Yamada, Yoshiya; Williamson, Matthew; Dauer, Lawrence T.; Zaider, Marco

    2014-03-01

    Purpose: A novel {sup 32}P brachytherapy source has been in use at our institution intraoperatively for temporary radiation therapy of the spinal dura and other localized tumors. We describe the dosimetry and clinical implementation of the source. Methods and Materials: Dosimetric evaluation for the source was done with a complete set of MCNP5 Monte Carlo calculations preceding clinical implementation. In addition, the depth dose curve and dose rate were measured by use of an electron field diode to verify the Monte Carlo calculations. Calibration procedures using the diode in a custom-designed phantom to provide an absolute dose calibration and to check dose uniformity across the source area for each source before treatment were established. Results: Good agreement was established between the Monte Carlo calculations and diode measurements. Quality assurance measurements results are provided for about 100 sources used to date. Clinical source calibrations were usually within 10% of manufacturer specifications. Procedures for safe handling of the source are described. Discussion: Clinical considerations for using the source are discussed.

  3. /sup 32/P-postlabeling assay in mice of transplacental DNA damage induced by the environmental carcinogens safrole, 4-aminobiphenyl, and benzo(a)pyrene

    SciTech Connect

    Lu, L.J.; Disher, R.M.; Reddy, M.V.; Randerath, K.

    1986-06-01

    Transplacental exposure of fetuses to carcinogens is known to induce tumors in the offspring, often with a high incidence and short latency. While covalent adduction of DNA appears to be essential for tumor initiation, little is known about the binding of carcinogens to the DNA of fetal tissues. A sensitive /sup 32/P-postlabeling method enabled us to study the binding of the environmental carcinogens safrole (600 mumol/kg p.o.), 4-aminobiphenyl (800 mumol/kg), and benzo(a)pyrene (200 mumol/kg) to the DNA of various maternal and fetal tissues after administration of test carcinogens to pregnant ICR mice on day 18 of gestation. The results show that these carcinogens bound to the DNA of maternal and fetal liver, lung, kidney, heart, brain, intestine, skin, maternal uterus, and placenta, with organ-specific quantitative and qualitative differences. It was possible for the first time to analyze DNA adduct patterns in minute amounts of tissue, for example those available from fetal heart. The covalent binding index 24 h after safrole treatment was estimated for the different organs and ranged from 0.1 to 247 and 0.1 to 5.8 for maternal and fetal DNA, respectively. Covalent binding index values of 0.2 to 13 and 0.1 to 0.3 for maternal and fetal DNA, respectively, were found for 4-aminobiphenyl. Benzo(a)pyrene treatment yielded covalent binding index values of 0.6 to 6.5 and 0.3 to 0.7 for maternal and fetal DNA, respectively. In both maternal and fetal tissues, safrole exhibited preferential binding to liver DNA. 4-Aminobiphenyl bound preferentially to DNA of maternal liver and kidney but showed no preference among fetal tissues. Benzo(a)pyrene exhibited weak tissue preference in both maternal and fetal organs.

  4. Identification of an interstitial 18p11.32-p11.31 duplication including the EMILIN2 gene in a family with porokeratosis of Mibelli.

    PubMed

    Occella, Corrado; Bleidl, Dario; Nozza, Paolo; Mascelli, Samantha; Raso, Alessandro; Gimelli, Giorgio; Gimelli, Stefania; Tassano, Elisa

    2013-01-01

    Porokeratosis is a rare disease of epidermal keratinization characterized by the histopathological feature of the cornoid lamella, a column of tightly fitted parakeratocytic cells, whose etiology is still unclear. Porokeratosis of Mibelli is a subtype of porokeratosis presenting a single plaque or a small number of plaques of variable size located unilaterally on limbs. It frequently appears in childhood and occurs with a higher incidence in males. Cytogenetic analyses were performed in all members of the family on lesioned and uninvolved skin. An array-CGH analysis was also performed utilizing the Human Genome CGH Microarray Kit G3 400 with 5.3 KB overall median probe spacing. Gene expression was performed on skin fibroblasts. In this study, we describe a Caucasian healthy 4-year-old child and his father showing features of porokeratosis of Mibelli. Array-CGH analysis revealed an interstitial 429.5 Kb duplication of chromosome 18p11.32-p11.3 containing four genes, namely: SMCHD1, EMILIN2, LPIN2, and MYOM1 both in patient and his father. EMILIN2 resulted overexpressed on skin fibroblasts. Also other members of this family, without evident signs of porokeratosis, carried the same duplication. Among these genes, we focused our attention on elastin microfibril interfacer 2 (EMILIN2) gene. Apoptosis plays a fundamental role in maintaining epidermal homeostasis, balancing keratinocytes proliferation, and forming the stratum corneum. EMILIN2 is known to trigger the apoptosis of different cell lines negatively affecting cell survival. It is expressed in the skin. We could speculate that the duplication and overexpression of EMILIN2 cause an abnormal apoptosis of epidermal keratinocytes and alter the process of keratinization, even if other epigenetic and genetic factors could also be involved. Our results could contribute to a better understanding of the pathogenesis of porokeratosis of Mibelli.

  5. Identification of an Interstitial 18p11.32-p11.31 Duplication Including the EMILIN2 Gene in a Family with Porokeratosis of Mibelli

    PubMed Central

    2013-01-01

    Porokeratosis is a rare disease of epidermal keratinization characterized by the histopathological feature of the cornoid lamella, a column of tightly fitted parakeratocytic cells, whose etiology is still unclear. Porokeratosis of Mibelli is a subtype of porokeratosis presenting a single plaque or a small number of plaques of variable size located unilaterally on limbs. It frequently appears in childhood and occurs with a higher incidence in males. Cytogenetic analyses were performed in all members of the family on lesioned and uninvolved skin. An array-CGH analysis was also performed utilizing the Human Genome CGH Microarray Kit G3 400 with 5.3 KB overall median probe spacing. Gene expression was performed on skin fibroblasts. In this study, we describe a Caucasian healthy 4-year-old child and his father showing features of porokeratosis of Mibelli. Array-CGH analysis revealed an interstitial 429.5 Kb duplication of chromosome 18p11.32-p11.3 containing four genes, namely: SMCHD1, EMILIN2, LPIN2, and MYOM1 both in patient and his father. EMILIN2 resulted overexpressed on skin fibroblasts. Also other members of this family, without evident signs of porokeratosis, carried the same duplication. Among these genes, we focused our attention on elastin microfibril interfacer 2 (EMILIN2) gene. Apoptosis plays a fundamental role in maintaining epidermal homeostasis, balancing keratinocytes proliferation, and forming the stratum corneum. EMILIN2 is known to trigger the apoptosis of different cell lines negatively affecting cell survival. It is expressed in the skin. We could speculate that the duplication and overexpression of EMILIN2 cause an abnormal apoptosis of epidermal keratinocytes and alter the process of keratinization, even if other epigenetic and genetic factors could also be involved. Our results could contribute to a better understanding of the pathogenesis of porokeratosis of Mibelli. PMID:23593459

  6. 2-Azido-( sup 32 P)NAD+, a photoactivatable probe for G-protein structure: Evidence for holotransducin oligomers in which the ADP-ribosylated carboxyl terminus of alpha interacts with both alpha and gamma subunits

    SciTech Connect

    Vaillancourt, R.R.; Dhanasekaran, N.; Johnson, G.L.; Ruoho, A.E. )

    1990-05-01

    A radioactive and photoactivatable derivative of NAD+, 2-azido-(adenylate-32P)NAD+, has been synthesized and used with pertussis toxin to ADP-ribosylate Cys347 of the alpha subunit (alpha T) of GT, the retinal guanine nucleotide-binding protein. ADP-ribosylation of alpha T followed by light activation of the azide moiety of 2-azido-(adenylate-32P)ADP-ribose produced four crosslinked species involving the alpha and gamma subunits of the GT heterotrimer: an alpha trimer (alpha-alpha-alpha), and alpha-alpha-gamma crosslink, an alpha dimer (alpha-alpha), and an alpha-gamma crosslink. The alpha trimer, alpha-alpha-gamma complex, alpha dimer, and alpha-gamma complexes were immunoreactive with alpha T antibodies. The alpha-alpha-gamma and the alpha-gamma complexes were immunoreactive with antisera recognizing gamma subunits. No evidence was found for crosslinking of alpha T to beta T subunits. Hydrolysis of the thioglycosidic bond between Cys347 and 2-azido-(adenylate-32P)ADP-ribose using mercuric acetate resulted in the transfer of radiolabel from Cys347 of alpha T in the crosslinked oligomers to alpha monomers, indicative of intermolecular photocrosslinking, and to gamma monomers, indicative of either intermolecular crosslinked complexes (between heterotrimers) or intramolecular crosslinked complexes (within the heterotrimer). These results demonstrate that GT exists as an oligomer and that ADP-ribosylated Cys347, which is four residues from the alpha T-carboxyl terminus, is oriented toward and in close proximity to the gamma subunit.

  7. Adjuvant treatment for early epithelial ovarian cancer: results of two randomised clinical trials comparing cisplatin to no further treatment or chromic phosphate (32P). G.I.C.O.G.: Gruppo Interregionale Collaborativo in Ginecologia Oncologica.

    PubMed

    Bolis, G; Colombo, N; Pecorelli, S; Torri, V; Marsoni, S; Bonazzi, C; Chiari, S; Favalli, G; Mangili, G; Presti, M

    1995-11-01

    From 1983 to 1990, 271 consecutive patients with stage I ovarian cancer entered two randomised trials, aimed at assessing the role of adjuvant chemotherapy after radical surgery in early stages of ovarian cancer. Trial I compared cisplatin (50 mg/m2 with repeated courses every 28 days for 6 cycles) to no further therapy in F.I.G.O. stage Ia & b Grade II-III patients; trial II compared cisplatin (same dose and schedule) to 32P in Iaii & bii and Ic patients. Both studies were multicentric and centrally randomized. Treatment was allocated by phone and stratified by center. All patients satisfying major eligibility criteria (histological and grade, no previous neoplasms) were analysed according to treatment allocated by randomisation. With a median observation time of 76 months, cisplatin significantly reduced the relapse rate by 65% (HR = 0.35; 95% CI = 0.14-0.89, p = 0.028; Cox Model) in trial I and 61% (HR = 0.39; 95% CI = 0.19-0.77, p = 0.007; Cox Model) in trial II. Survival was not significantly different (trial I - Kaplan-Meier overall 5-year survival: cisplatin = 88%, control = 82%, HR = 1.15; 95% CI = 0.44-2.98; p = 0.773; Cox Model); trial II - overall 5-year survival: cisplatin = 81%, 32P = 79%, HR = 0.72; 95% CI = 0.37-1.43; p = 0.354; Cox model). In both studies the risk of dying after relapse increased for patients originally randomized to the cisplatin arms: in trial I, 6 of 7 patients in the cisplatin relapsed arm and died of tumor compared with 8 of 14 patients in the control arm. In trial II 11 of 12 patients on cisplatin, and 18 of 26 on 32P succumbed to tumor recurrence. Adjuvant cisplatin treatment in early ovarian cancer significantly prevents relapse in comparison to 32P in stage IC patients or to no immediate treatment in earlier stage women. The impact of cisplatin adjuvant treatment on survival remains, however, unclear.

  8. N-(2-chloroethyl)-N-nitrosoureas covalently bound to nonionic and monocationic lexitropsin dipeptides. Synthesis, DNA affinity binding characteristics, and reactions with sup 32 P-end-labeled DNA

    SciTech Connect

    Church, K.M.; Wurdeman, R.L.; Zhang, Yi; Chen, Faxian; Gold, B. )

    1990-07-24

    The synthesis and characterization of a series of compounds that contain an N-alkyl-N-nitrosourea functionality linked to DNA minor groove binding bi- and tripeptides (lexitropsins or information-reading peptides) based on methylpyrrole-2-carboxamide subunits are described. The lexitropsins (lex) synthesized have either a 3-(dimethylamino)propyl or propyl substituent on the carboxyl terminus. The preferred DNA affinity binding sequences of these compounds were footprinted in {sup 32}P-end-labeled restriction fragments with methidiumpropyl-EDTA{center dot}Fe(II), and in common with other structural analogues, e.g., distamycin and netropsin, these nitrosoureas recognize A-T-rich runs. The affinity binding of the compound with the dimethylamino terminus, which is ionized at near-neutral pH, appeared stronger than that observed for the neutral dipeptide. The sequence specificity for DNA alkylation by (2-chloroethyl)nitrosourea-lex dipeptides (Cl-ENU-lex), with neutral and charged carboxyl termini, using {sup 32}P-end-labeled restriction fragments, was determined by the conversion of the adducted sites into single-strand breaks by sequential heating at neutral pH and exposure to base. The DNA cleavage sites were visualized by polyacrylamide gel electrophoresis and autoradiography. Linking the Cl-ENU moiety to minor groove binders is a viable strategy to qualitatively and quantitatively control the delivery and release of the ultimate DNA alkylating agent in a sequence-dependent fashion.

  9. Distribution of /sup 32/P in laboratory colonies of Solenopsis invicta (Hymenoptera: Formicidae) after feeding on labeled Heliothis zeal (Lepidoptera: Noctuidae) eggs: an explanation of discrepancies encountered in field predation experiments

    SciTech Connect

    Nuessly, G.S.; Sterling, W.L.

    1986-12-01

    Factors responsible for low recovery rates of radioactive Solenopsis invicta Buren following placement of /sup 32/P-labeled Heliothis zea (Boddie) eggs on cotton in field predation tests were investigated using laboratory colonies of the ants. S. invicta workers became radioactive while handling labeled eggs by rupturing the egg chorion or by picking up labeled substances present on the surface of eggs. Foragers that removed the eggs from the plants picked up significantly more of the label than did workers that were sampled from the colonies between 12 and 72 h after egg introduction. Percentage of workers that became labeled over time was much lower with the solid live food than in other studies that used powdered food sources. Problems in finding labeled ants in the field may have been associated with low mean levels of /sup 32/P per ant, together with difficulty in locating and isolating labeled ants from the population. Results indicate that egg predation rates estimated from counts per minute per predator have high variability, and suggest fairly large errors in estimates of eggs consumed per ant. Use of recovery rates of labeled predators to improve estimation of predation rates is discussed.

  10. Analysis of the metabolic turnover of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate. Validation of novel analytical techniques by using 32P-labelled lipids from erythrocytes.

    PubMed Central

    Hawkins, P T; Michell, R H; Kirk, C J

    1984-01-01

    We have developed methods that yield estimates of the 32P content of each of the individual phosphate groups of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate, thus extending the information available from studies of the labelling of these lipids in intact cells or membrane preparations. The analyses are undertaken with the deacylated lipids. Assay of the 5-phosphate of phosphatidylinositol 4,5-bisphosphate is achieved by the use, under conditions of first-order kinetics, of a 5-phosphate-specific phosphomonoesterase present in isolated erythrocyte membranes [Downes, Mussat & Michell (1982) Biochem. J. 203, 169-177]. Assay of the 4-phosphate of phosphatidylinositol 4-phosphate and of the total monoester phosphate content (4-phosphate plus 5-phosphate) of phosphatidylinositol 4,5-bisphosphate employs alkaline phosphatase from bovine intestine. The radioactivity of the 1-phosphate is that remaining as organic phosphate after exhaustive alkaline phosphatase treatment. The methodology has been validated by using lipids from human erythrocytes: these contain no 32P in their 1-phosphate. These methods should be of substantial value in studies of the many cells that show rapid hormonal perturbations of phosphatidylinositol 4,5-bisphosphate metabolism. PMID:6326746

  11. Bis(trifluoromethyl)phosphinous acid (CF3)2P-O-H: an example of a thermally stable phosphinous acid--synthesis, gas-phase structure, and rotational isomers.

    PubMed

    Hoge, Berthold; Garcia, Placido; Willner, Helge; Oberhammer, Heinz

    2006-04-24

    The bis(trifluoromethyl)phosphinous acid, (CF3)2P-O-H, is the only known example of a thermally stable phosphinous acid. Although this compound has been known since 1960, little is known about the chemistry of this extraordinary compound; this might be due to the tedious, and in some part risky, synthesis that was originally published. An improved, simple, and safe synthesis that is based on the treatment of the easily accessible (CF3)2PNEt2, with at least three equivalents of p-toluene sulfonic acid, is presented. The reaction results in a complete conversion to the phosphinous acid, which is isolated in almost 90 % yield. The compound exists in an equilibrium of two P--OH rotational isomers, a fact which is supported by quantum chemical calculations. The relative enthalpy difference of 6.4 kJ mol(-1), calculated at the B3PW91/6-311G(3d,p) level, is in excellent agreement with the experimental value of 5.9 kJ mol(-1), which was determined from the temperature dependence of the nu(OH) bands of the two rotational isomers. The complete experimental vibrational spectra of both rotamers, their predicted vibrational spectra obtained by using quantum chemical calculations, and an attempt at photoinduced isomerization of matrix-isolated (CF3)2POH is presented. The experimental structure, obtained from an electron-diffraction study in the gas phase, is reproduced very well by ab initio and density functional theory (DFT) methods.

  12. Comparative DNA binding of 7,12-dimethylbenz[a]anthracene and some of its metabolites in mouse epidermis in vivo as revealed by the 32P-postlabeling technique.

    PubMed

    Schoepe, K B; Friesel, H; Schurdak, M E; Randerath, K; Hecker, E

    1986-04-01

    The binding of some mouse skin metabolites and related derivatives of the tumor initiator 7,12-dimethylbenz[a]anthracene (DMBA) was investigated by 32P-postlabeling analysis after its topical administration. DMBA and trans-3,4-dihydro-3,4-dihydroxy-DMBA (DMBA-3,4-dihydrodiol) both led to the formation of four DNA adducts, which showed a very similar pattern of spots on thin-layer chromatograms. With trans-8,9-dihydro-8,9-dihydroxy-7,12-dimethylbenz[a]anthracene (DMBA-8,9-dihydrodiol) one major adduct was obtained which was chromatographically indistinguishable from one of the DMBA adducts. In contrast, 7-hydroxymethyl-12-methylbenz[a]anthracene (7-OHM-12-MBA) gave rise to two major adducts which were separable from DMBA adducts. 3-hydroxy-7,12-dimethylbenz[a]anthracene (3-OH-DMBA) and 7,12-dimethylbenz[a]anthracene-7,12-epoxide (DMBA-O2) did not lead to detectable amounts of adducts. Quantitative determination of DNA binding showed that an initiating dose (i = 100 nmol) of DMBA yielded approximately 12 adducts/10(7) normal nucleotides. Adduct formation with the same dose of DMBA-3,4-dihydrodiol was 7-8 times higher. At a 4-fold higher dose level, DMBA-8,9-dihydrodiol exhibited a 3- to 6-times weaker binding and 7-OHM-12-MBA a slightly stronger binding than DMBA. Chromatography of the DMBA and DMBA-3,4-dihydrodiol adducts with a solvent containing borate showed a decreased mobility of two out of four adducts in each case. These adducts were also sensitive to oxidation by periodate. The results suggest that two DMBA adducts carried vicinal cis-hydroxyl groups and thus were probably derived from the anti-3,4-dihydrodiol-1,2-oxide(s) of DMBA. The other two adducts were probably derived from the syn-stereoisomer(s). When the DNA-modifying capabilities and initiating activities of the more prominent mouse-skin metabolites are considered in relation to DMBA, DMBA-3,4-dihydrodiol is postulated to be a proximate and DMBA-3,4-dihydrodiol-1,2-oxide(s) to be ultimate

  13. A method to accurately quantitate intensities of (32)P-DNA bands when multiple bands appear in a single lane of a gel is used to study dNTP insertion opposite a benzo[a]pyrene-dG adduct by Sulfolobus DNA polymerases Dpo4 and Dbh.

    PubMed

    Sholder, Gabriel; Loechler, Edward L

    2015-01-01

    Quantitating relative (32)P-band intensity in gels is desired, e.g., to study primer-extension kinetics of DNA polymerases (DNAPs). Following imaging, multiple (32)P-bands are often present in lanes. Though individual bands appear by eye to be simple and well-resolved, scanning reveals they are actually skewed-Gaussian in shape and neighboring bands are overlapping, which complicates quantitation, because slower migrating bands often have considerable contributions from the trailing edges of faster migrating bands. A method is described to accurately quantitate adjacent (32)P-bands, which relies on having a standard: a simple skewed-Gaussian curve from an analogous pure, single-component band (e.g., primer alone). This single-component scan/curve is superimposed on its corresponding band in an experimentally determined scan/curve containing multiple bands (e.g., generated in a primer-extension reaction); intensity exceeding the single-component scan/curve is attributed to other components (e.g., insertion products). Relative areas/intensities are determined via pixel analysis, from which relative molarity of components is computed. Common software is used. Commonly used alternative methods (e.g., drawing boxes around bands) are shown to be less accurate. Our method was used to study kinetics of dNTP primer-extension opposite a benzo[a]pyrene-N(2)-dG-adduct with four DNAPs, including Sulfolobus solfataricus Dpo4 and Sulfolobus acidocaldarius Dbh. Vmax/Km is similar for correct dCTP insertion with Dpo4 and Dbh. Compared to Dpo4, Dbh misinsertion is slower for dATP (∼20-fold), dGTP (∼110-fold) and dTTP (∼6-fold), due to decreases in Vmax. These findings provide support that Dbh is in the same Y-Family DNAP class as eukaryotic DNAP κ and bacterial DNAP IV, which accurately bypass N(2)-dG adducts, as well as establish the scan-method described herein as an accurate method to quantitate relative intensity of overlapping bands in a single lane, whether generated

  14. Adsorption of the cis-[Pt(NH3)2(P2O7)](2-) (phosphaplatin) on hydroxyapatite nanocrystals as a smart way to selectively release activated cis-[Pt(NH3)2Cl2] (cisplatin) in tumor tissues.

    PubMed

    Benedetti, Michele; De Castro, Federica; Romano, Alessandro; Migoni, Danilo; Piccinni, Barbara; Verri, Tiziano; Lelli, Marco; Roveri, Norberto; Fanizzi, Francesco P

    2016-04-01

    The relevant adsorption of cis-[Pt(NH3)2(P2O7)](2-) (phosphaplatin) on hydroxyapatite nanocrystals (nHAP) was observed and studied in water suspension. Phosphaplatin cytotoxicity, which is very low for HeLa, MCF-7 and HS-5 cell lines could be enhanced, reaching that of cisplatin, by interaction with solid nHAP. This effect stems from nHAP ability to catalyze the phosphaplatin hydrolysis, producing the same hydrolytic species responsible for cisplatin antitumor activity.

  15. Mitochondria with impaired phosphate transport: 32P uptake studies.

    PubMed

    Williams, G R; Orr, J L

    1976-02-01

    Rat liver mitochondria which have been exposed to 0.15 M NaC1 at 35 degrees C for 15 min subsequently take up 32Pi from an external medium only to about 5% of the extent of uptake by control mitochondria. The volume into which 32Pi distributes in a pellet of such "aged" mitochondia is less than that available to 3H2O but is greater than that available to [3H]sucrose. Mitochondria treated in this manner cannot therefore accumulate Pi although limited penetration of the inner membrane can occur. These results confirm earlier findings by indirect methods (Williams, G. R. & Orr, J. L.: Dynamics of energy-transducing membranes (Ernster, L., Estabrook, R. W. & Slater, E. C., eds), Elsevier Scientific Publishing Company, Amsterdam, Netherlands, pp. 497-508 (1974).

  16. [Relations between location of elements in periodic table and affinity for the kidneys (author's transl)].

    PubMed

    Ando, A; Hisada, K; Ando, I

    1977-10-01

    The distribution of many inorganic compounds in rats was investigated in order to evaluate kidney affinity of inorganic compounds. In these experiments, 30%, 10-20% and 4-10% of administered dose was localized in the kidneys in 203Hg-acetate and 203 Bi-acetate, in H198AuCl4, 103PdCl2, 201TlCl, 210Pd(NO3)2 and H2(127M)TeO3, and in Na2(51)CrO4, 54MnCl2, (114m)InCl3 and 7BeCl2, respectively. Some bipositive ions and anions was hardly taken up into the kidneys. And in many hard acids according to classification of Lewis acids, the uptake rate into the kidneys was usually small. On the other hand, Hg, Au and Bi, which have strong binding power to the protein, showed high uptake rate in the kidneys. As Hg++, Au+ and Bi+++ was soft acids according to classification of Lewis acids, it was thought that these elements would bind strongly to soft base (RSH, RS-) present in the kidney.

  17. Internal irradiation for cystic craniopharyngioma

    SciTech Connect

    Kobayashi, T.; Kageyama, N.; Ohara, K.

    1981-12-01

    The authors report the results of internal irradiation with labeled chromic phosphate (32P) and gold-198 (198Au) colloid in eight cases of cystic craniopharyngiomas. They used a newly developed dosimetric formula, by which the radiation dose at the cyst wall and at any point far from the radioactive source can be calculated. Ten courses of irradiation in eight patients were carried out by injection of either 32P or 198Au colloid into the cyst through an Ommaya drainage system that had been placed at craniotomy. Follow-up studies ranging from 13 to 156 months revealed that all cysts were effectively treated, with elimination of fluid or collapse of the cyst. This was confirmed by Conray cystography and/or computerized tomography. Not only the dose delivered to the wall but also the thickness of the cyst wall and the location of the cyst are important factors in planning internal irradiation. A safe and adequate dose to the cyst wall could range between 9000 to 30,000 rads for craniopharyngioma. This treatment is suitable for large cysts that are thought to be difficult to remove radically, recurrent cysts resistant to previous treatment, or multiple cysts. Internal irradiation may also be applicable in other cystic intracranial tumors if dosimetry is calculated accurately.

  18. Establishment of a trimodality analytical platform for tracing, imaging and quantification of gold nanoparticles in animals by radiotracer techniques.

    PubMed

    Chen, Chien-Hung; Lin, Fong-Sian; Liao, Wei-Neng; Liang, Sanching L; Chen, Min-Hua; Chen, Yo-Wen; Lin, Wan-Yu; Hsu, Ming-Hua; Wang, Mei-Ya; Peir, Jinn-Jer; Chou, Fong-In; Chen, Ching-Ya; Chen, Sih-Yu; Huang, Su-Chin; Yang, Mo-Hsiung; Hueng, Dueng-Yuan; Hwu, Yeukuang; Yang, Chung-Shi; Chen, Jen-Kun

    2015-01-06

    This study aims to establish a (198)Au-radiotracer technique for in vivo tracing, rapid quantification, and ex vivo visualization of PEGylated gold nanoparticles (GNPs) in animals, organs and tissue dissections. The advantages of GNPs lie in its superior optical property, biocompatibility and versatile conjugation chemistry, which are promising to develop diagnostic probes and drug delivery systems. (198)Au is used as a radiotracer because it simultaneously emits beta and gamma radiations with proper energy and half-life; therefore, (198)Au can be used for bioanalytical purposes. The (198)Au-tagged radioactive gold nanoparticles ((198)Au-GNPs) were prepared simply by irradiating the GNPs in a nuclear reactor through the (197)Au(n,γ)(198)Au reaction and subsequently the (198)Au-GNPs were subjected to surface modification with polyethylene glycol to form PEGylated (198)Au-GNPs. The (198)Au-GNPs retained physicochemical properties that were the same as those of GNP before neutron irradiation. Pharmacokinetic and biodisposition studies were performed by intravenously injecting three types of (198)Au-GNPs with or without PEGylation into mice; the γ radiation in blood specimens and dissected organs was then measured. The (198)Au-radiotracer technique enables rapid quantification freed from tedious sample preparation and shows more than 95% recovery of injected GNPs. Clinical gamma scintigraphy was proved feasible to explore spatial- and temporal-resolved biodisposition of (198)Au-GNPs in living animals. Moreover, autoradiography, which recorded beta particles from (198)Au, enabled visualizing the heterogeneous biodisposition of (198)Au-GNPs in different microenvironments and tissues. In this study, the (198)Au-radiotracer technique facilitated creating a trimodality analytical platform for tracing, quantifying and imaging GNPs in animals.

  19. [Metabolism and excretion of 32P-aminophon in lactating cattle].

    PubMed

    Dedek, W; Grahl, R; Schwarz, H

    1978-01-01

    P-labelled aminophon, 0,0-di-u-butyl- (1-n-butylaminocyclohexyl) -phosphonate, an agricultural defoliant and siccant, was applied orally in oily solution to lactating cows, 5-6 mg/kg bodymass, resp. The halflifes of degradation in blood serum in vitro are 95 min, of the extractable metabolites in blood, milk and urine 17-20 h. The 0-and 0, N-dealkylcompound of aminophon were found as the preferred metabolites.

  20. Materials Data on Bi32P10Pb3O76 (SG:5) by Materials Project

    SciTech Connect

    Kristin Persson

    2016-02-04

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  1. Detection of Sulfur Mustard-Induced DNA Damage by a 32P-Postlabeling Method

    DTIC Science & Technology

    1993-05-13

    of the method depends on developing separation techniques which allow the adduct to be measured in the presence of much higher concentrations of...2PIATP. | - CONCLUSIONS The 3 2P-postlabeling technique has been applied successfully to the detection of HETEG, the major adduct produced in DNA by...Pharmacology0. University of Massachusetts Medical School W orcester, MA 01655. ABSTRACT We have applied the 3 2 P-postlabeling technique to the detection of

  2. gamma-Glutamyltransferase-dependent biliary-hepatic recycling of methyl mercury in the guinea pig.

    PubMed

    Dutczak, W J; Ballatori, N

    1992-08-01

    After secretion into bile, glutathione (GSH) and GSH-conjugates are catabolized by gamma-glutamyltransferase (gamma-GT) and dipeptidases yielding glutamate, cysteine, glycine and cysteine S-conjugates, and these products are then partially reabsorbed from the biliary tree. Because methyl mercury is thought to be secreted into bile as a GSH- complex, it may be subject to a similar intrahepatic cycle, thus delaying its elimination. To examine this possibility guinea pigs were dosed with 203Hg-methyl mercury (10 mumol/kg i.v.), followed by a retrograde intrabiliary infusion of Krebs-Henseleit buffer (control) or acivicin (an inhibitor of gamma-GT). Acivicin increased biliary excretion of 203Hg by 41%, and GSH from 0.14 +/- 0.10 to 2.02 +/- 0.26 nmol/min.g of liver. Bile analyzed by gel filtration chromatography revealed that CH(3)203Hg-GSH accounted for most of this increased 203Hg excretion. When CH(3)203Hg-complexes of GSH, cysteine and albumin were introduced directly into the biliary tree by retrograde infusion, 203Hg recovery in bile was significantly lower than recovery of the nonabsorbable marker [14C]sucrose, ranging from 26.0 +/- 2.9% for CH(3)203Hg-cysteine to 48.7 +/- 5.1% for CH(3)203Hg-albumin and approximately 60% for [14C]sucrose. Acivicin pretreatment significantly increased 203Hg excretion into bile after retrograde infusion of CH(3)203Hg-GSH, whereas 203Hg recovery after retrograde infusion of CH(3)203Hg-cysteine remained constant.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Effects of acetylcholine and other agents on /sup 32/P-prelabeled phosphoinositides and phosphatidate in crude synaptosomal preparations

    SciTech Connect

    White, H.L.

    1988-05-01

    Experimental conditions are described which permit effects of various agents on polyphosphoinositides and phosphatidic acid (PA) to be evaluated simultaneously in crude nerve-ending preparations from rat brain. Acetylcholine (3-100 microM) or carbachol (30-1,000 microM) induced the hydrolysis of prelabeled polyphosphoinositides and, at the same time, stimulated the net label incorporated in phosphatidic acid. All muscarinic effects were blocked by atropine or pirenzepine. Non-muscarinic agonists (glutamate, adenosine, norepinephrine) stimulated polyphosphoinositide hydrolysis in this preparation, but of these only norepinephrine affected phosphatidic acid turnover. A potentiation of acetylcholine-induced phosphoinositide turnover by KCl was observed, as well as an apparent selective inhibition of PIP2 hydrolysis by LiCl. Acetylcholine-stimulated turnover of PA was not necessarily coupled to phosphoinositide hydrolysis.

  4. Cosmogenic 32P and 33P in the Atmosphere and Oligotrophic Ocean and Applications to the Study of Phosphorus Cycling

    DTIC Science & Technology

    1993-02-01

    Woods Hc’e Oceanographic Institution Joint Program. 3 Reproduction in whole or in part is permitted for any purpos of the United States Government. This...al., 1961). Also it has been shown that zooplankton have the ability to store P as fat in the males and in the reproductive system in the females... sargassum . I Table 4.4. Specific activities of 3 2 P and 3 3 P and activity ratio 3 3 p/ 2 p in plankton tows and suspended particulate (> 3 pmo

  5. Bioavailability of methylmercury to Sacramento blackfish (Orthodon microlepidotus): Dissolved organic carbon effects

    SciTech Connect

    Choi, M.H.; Cech, J.J. Jr.; Lagunas-Solar, M.C.

    1998-04-01

    The effect of dissolved organic carbon (DOC) on methylmercury (MeHg) uptake across the gills of Sacramento blackfish (Orthodon microlepidotus) was investigated using the Hg-203 radioisotope. The efficiency of fish gills in extracting MeHg from water was measured using a McKim-type fish respirometer that separated exposure water from expired water. Blackfish gill ventilation and oxygen consumption rates remained constant, while Me{sup 203}Hg uptake was decreased significantly in the presence of DOC. Mean Me{sup 203}Hg extraction efficiency, uptake rate constant, and blood to inspired water ratio decreased 78%, 73%, and 63%, respectively, with 2 mg C/L of DOC, and 85%, 82%, and 70% with 5 mg C/L DOC, compared to the Me{sup 203}Hg reference treatment group. Because respiratory parameters remained unchanged, reductions in Me{sup 203}Hg uptake indicate strong interactions between DOC and Me{sup 203}Hg Methyl{sup 203}Hg levels in fish gills, kidney, and spleen from 2 and 5 mg C/L were significantly lower than those observed from the reference treatment group. These reductions in uptake (bioavailability) support the hypothesis that trans-gill transport of Me{sup 203}Hg is inhibited when it is complexed by DOC in the aqueous medium, decreasing Me{sup 203}Hg uptake and accumulation in fish organs.

  6. Plasmidless, photosynthetically incompetent mutants of Rhodospirillum rubrum.

    PubMed Central

    Kuhl, S A; Wimer, L T; Yoch, D C

    1984-01-01

    Ethyl methanesulfonate rendered a high percentage of Rhodospirillum rubrum cells plasmidless and photosynthetically incompetent (Kuhl et al., J. Bacteriol. 156:737-742, 1983). By probing restriction endonuclease-digested chromosomal DNA from these plasmidless strains with 32P-labeled R. rubrum plasmid DNA, we showed that no homology exists between the plasmid and the chromosomal DNA of the mutant. Loss of the plasmid in all the nonphotosynthetic isolates was accompanied by the synthesis of spirilloxanthin under aerobic growth conditions, resistance to cycloserine and HgCl2, and loss of ability to grow fermentatively on fructose. Changes in both the protein and lipid composition of the membranes and the impaired uptake of 203HgCl2 in the plasmidless strains (compared with the wild type) suggest either that membrane modification occurs as a result of plasmid loss, accounting for several of the acquired phenotype characteristics of the cured strains, or that both membrane modification and plasmid loss are part of the same pleiotropic mutation. Images PMID:6434514

  7. Response of phage T4 polynucleotide kinase toward dinucleotides containing apurinic sites: Design of a sup 32 P-postlabeling assay for apurinic sites in DNA

    SciTech Connect

    Weinfeld, M.; Liuzzi, M.; Paterson, M.C. )

    1990-02-20

    The authors have examined the capacity of bacteriophage T4 polynucleotide kinase to phosphorylate the partially depurinated products of d-ApA, namely d-SpA and d-ApS (where S represents an apurinic deoxyribose group). It was observed that the enzyme acted only on the latter isomer. Since molecules of this type (d-NpS) are the sole apurinic site containing products resulting from the combined digestion of lightly depurinated DNA by snake venom phosphodiesterase and calf alkaline phosphatase they were able to devise a postlabeling assay for these biologically important DNA lesions. The method offers several advantages, including (a) elimination of the need for prelabeled DNA, (b) high (femtomole range) sensitivity, and (c) nearest-neighbor analysis of bases 5{prime} to apurinic/apyrimidinic sites. Using this assay, they obtained a value for the rate of depurination of form I pRSV neo plasmid DNA. The rate of depurination of poly(dA), treated in a similar fashion, was found to be {approximately}1 base per 10{sup 3} nucleotides per hour.

  8. Cloning of a New Gene/s in Chromosome 17p3.2-p13.1 that Control Apoptosis

    DTIC Science & Technology

    2005-04-01

    Sequence Detection System, December 11, 1997, updated 10/2001). B-III-i- Apoptosis assays. The cells were washed with cold Guava Nexin Buffer and stained...in a 50pl reaction volume with Guava Nexin PE and Guava Nexin 7-AAD. The stained cells were diluted to 500 [1l with cold Guava Nexin Buffer and...acquired on the Guava PC. Data was acquired on the using Guava PCA system using CytoSoft software. The cells were induced to undergo apoptosis by incubation

  9. Production and modeling of radioactive gold nanoparticles in Tehran research reactor.

    PubMed

    Hosseini, Seyedeh Fatemeh; Sadeghi, Mahdi; Aboudzadeh, Mohammad Reza; Mohseni, Morteza

    2016-12-01

    Gold has two medically useful radioactive isotopes, (198)Au and (199)Au, for locally irradiating and killing tumor cells. (198)Au radionuclide has been produced through the irradiation of the pure gold via (197)Au(n,γ)(198)Au reaction in the Tehran Research Reactor at a thermal neutron flux of 4.5×10(13)ncm(-2)s(-1) for the different irradiation times. In this paper, the activity of (198)Au radionuclide has been determined using MCNPX-2.6 and TALYS-1.6 codes and also the theoretical approach. The calculated results were compared with the corresponding experimental values. The calculated results were in good agreement with the experimental data, thus the used codes can be used as a powerful tool to predict and optimize production conditions in reactor.

  10. Development of a Short-Lived Radioisotope Production Service (SRPS) for CTTC at the University of Alberta SLOWPOKE Reactor Facility

    DTIC Science & Technology

    2004-12-01

    half-life the radioisotopes include : 165Dy, 56~ 171Er, 42~ 64Cu, 72Ga, 142Pr, 24Na, 187W, 7 6 As, 166Ho, 82Br, 140La, 153Sm, and 198 Au. Of...As, 142Pr, 166Ho, 198 Au, 165Dy and 56Mn would be of the order of$1800- $220 0 per radioisotope. The cost of producing sufficient activity t o have...1.4708 d La 139La (n,y) t4oLa 14.4 533 1.6781 d Sm 137.8 5098 1.9285 d u (Np) 13.3 491 239Np(U) 2.355 d Au 197 Au (n,y) 198 Au 142.5 52 71 198Au

  11. IEA-R1 Nuclear Research Reactor: 58 Years of Operating Experience and Utilization for Research, Teaching and Radioisotopes Production

    SciTech Connect

    Cardenas, Jose Patricio Nahuel; Filho, Tufic Madi; Saxena, Rajendra; Filho, Walter Ricci

    2015-07-01

    IEA-R1 research reactor at the Instituto de Pesquisas Energeticas e Nucleares (Nuclear and Energy Research Institute) IPEN, Sao Paulo, Brazil is the largest power research reactor in Brazil, with a maximum power rating of 5 MWth. It is being used for basic and applied research in the nuclear and neutron related sciences, for the production of radioisotopes for medical and industrial applications, and for providing services of neutron activation analysis, real time neutron radiography, and neutron transmutation doping of silicon. IEA-R1 is a swimming pool reactor, with light water as the coolant and moderator, and graphite and beryllium as reflectors. The reactor was commissioned on September 16, 1957 and achieved its first criticality. It is currently operating at 4.5 MWth with a 60-hour cycle per week. In the early sixties, IPEN produced {sup 131}I, {sup 32}P, {sup 198}Au, {sup 24}Na, {sup 35}S, {sup 51}Cr and labeled compounds for medical use. During the past several years, a concerted effort has been made in order to upgrade the reactor power to 5 MWth through refurbishment and modernization programs. One of the reasons for this decision was to produce {sup 99}Mo at IPEN. The reactor cycle will be gradually increased to 120 hours per week continuous operation. It is anticipated that these programs will assure the safe and sustainable operation of the IEA-R1 reactor for several more years, to produce important primary radioisotopes {sup 99}Mo, {sup 125}I, {sup 131}I, {sup 153}Sm and {sup 192}Ir. Currently, all aspects of dealing with fuel element fabrication, fuel transportation, isotope processing, and spent fuel storage are handled by IPEN at the site. The reactor modernization program is slated for completion by 2015. This paper describes 58 years of operating experience and utilization of the IEA-R1 research reactor for research, teaching and radioisotopes production. (authors)

  12. In vitro radioisotopic labeling of the L-3 stage of Brugia malayi.

    PubMed

    Lengemann, F W; Marchell, T F; Chmielewicz, M; Georgi, J R

    1986-07-01

    Ten radioisotopes (133Ba, 207Bi, 109Cd, 51Cr, 59Fe, 203Hg, 54Mn, 125Sb and 75Se) were used to label the L-3 stage of Brugia malayi. The in vitro uptake at 4 hours in a buffered salt solution was greatest for 207Bi and exceeded that of 203Hg, by a factor of 6 and the rest of the radionuclides by factors of 26 or more. Dimethylsulfoxide (DMSO) (1%) in the medium only slightly enhanced the uptake of 207Bi but increased 203Hg uptake by a factor of 2.5. After incubation in non-radioactive medium only 2% of the 207Bi was retained; for 203Hg the retention was 70% or better. Increasing the labeling time increased total uptake and retention of the radionuclides. The retention of the 203Hg injected intraperitoneally into jirds (Meriones unguiculatus) in the form of labeled L-3s was followed over a 16 day period by counting the jirds in a whole animal NaI crystal detector. One L-4 stage of B malayi was recovered at the end of this period.

  13. In vitro radioisotopic labeling of the L-3 stage of Brugia malayi

    SciTech Connect

    Lengemann, F.W.; Marchell, T.F.; Chmielewicz, M.; Georgi, J.R.

    1986-07-01

    Ten radioisotopes (/sup 133/Ba, /sup 207/Bi, /sup 109/Cd, /sup 51/Cr, /sup 59/Fe, /sup 203/Hg, /sup 54/Mn, /sup 125/Sb and /sup 75/Se) were used to label the L-3 stage of Brugia malayi. The in vitro uptake at 4 hours in a buffered salt solution was greatest for /sup 207/Bi and exceeded that of /sup 203/Hg, by a factor of 6 and the rest of the radionuclides by factors of 26 or more. Dimethylsulfoxide (DMSO) (1%) in the medium only slightly enhanced the uptake of /sup 207/Bi but increased /sup 203/Hg uptake by a factor of 2.5. After incubation in non-radioactive medium only 2% of the /sup 207/Bi was retained; for /sup 203/Hg the retention was 70% or better. Increasing the labeling time increased total uptake and retention of the radionuclides. The retention of the /sup 203/Hg injected intraperitoneally into jirds (Meriones unguiculatus) in the form of labeled L-3s was followed over a 16 day period by counting the jirds in a whole animal NaI crystal detector. One L-4 stage of B malayi was recovered at the end of this period.

  14. Cyclotron produced 198gAu, a potential radionuclide for diagnostic and therapeutic applications

    NASA Astrophysics Data System (ADS)

    Khandaker, Mayeen Uddin; Haba, Hiromitsu; Kassim, Hasan Abu

    2016-02-01

    Production cross-sections of the natPt(d,x)198Au reactions have been measured from a 24-MeV deuteron energy down to the threshold by using a stacked-foil activation technique combined with HPGe γ-ray spectrometry. Only a partial agreement is obtained with the existing literature data and the theoretical data extracted from the TENDL-2013 library. Physical thick target yield for the 198Au radionuclide was deduced using the measured cross-sections, and found a general agreement with the directly measured yield available in the literature. This study reveals that a low deuteron energy (<15 MeV) cyclotron and an enriched 198Pt (100%) target could be used to obtain 198Au in no carrier added form.

  15. Effects of substituted dithiocarbamates on distribution and excretion of inorganic mercury in mice

    SciTech Connect

    Gale, G.R.; Atkins, L.M.; Smith, A.B.; Jones, M.M.

    1985-02-01

    Diethyldithiocarbamate (DDTC) and six of its N,N-disubstituted congeners were evaluated for their relative efficacies in mobilizing and promoting excretion of mercury (Hg/sup + +/) using mice which had been given /sup 203/Hg/sup + +/; D-penicillamine (PEN) was used as a positive control compound. While none was as effective as PEN when compared on a molar dose basis, significant activity as assessed by reduction of whole body /sup 203/Hg burden, was observed following treatment with three of the analogs. The order of effectiveness was PEN greater than N-methyl-N-dithiocarboxyglucamine (MDCG) greater than di(carboxymethyl)dithiocarbamate (DCDC) greater than di(hydroxyethyl)dithiocarbamate (DHDC).

  16. A rare prenatal case with two de novo inversions and a translocation: 48, XX,t(9;12)(q32;p24.3), inv(11)(p15.1q25), inv(13)(q12.q22)

    SciTech Connect

    Harrison, B.; Balaban, L.; Eldred, C.

    1994-09-01

    Ultrasound examination of a para 1, gravida 2, 26 y.o. showed severe hydrocephalus and polyhydramnios. Amniocentesis was performed at 27 weeks. High resolution chromosome analysis revealed a karyotype with a 9;12 translocation, a pericentric inversion of chromosome 11, and a paracentric inversion of chromosome 13. Parental chromosome studies were normal. The mother was not on medication prior to her pregnancy and there was no known exposure to radiation. Delivery was at 34 weeks gestation. The phenotype consisted of micrognathia, low set ears, hypertelorism, and hydrodcephaly. Review of the literature revealed a single report with multiple de novo aberrations consisting of a 6;14 translocation and a deleted 7. This was diagnosed in the child of a woman with systemic lupus erythematous treated with azathioprine. These types of abnormalities have been known to be induced by chemical and radiation exposure. High resolution banding combined with molecular studies presently improve our ability to detect subtle structural aberrations.

  17. 10 CFR Appendix L to Part 110 - Illustrative List of Byproduct Materials Under NRC Export/Import Licensing Authority a

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...) Gadolinium 153 (Gd 153) Gadolinium 159 (Gd 159) Gallium 67 (Ga 67) Gallium 72 (Ga 72) Germanium 68 (Ge 68) Germanium 71 (Ge 71) Gold 195 (Au 195) Gold 198 (Au 198) Gold 199 (Au 199) Hafnium 172 (Hf 172) Hafnium 181...

  18. 10 CFR Appendix L to Part 110 - Illustrative List of Byproduct Materials Under NRC Export/Import Licensing Authority a

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...) Gadolinium 153 (Gd 153) Gadolinium 159 (Gd 159) Gallium 67 (Ga 67) Gallium 72 (Ga 72) Germanium 68 (Ge 68) Germanium 71 (Ge 71) Gold 195 (Au 195) Gold 198 (Au 198) Gold 199 (Au 199) Hafnium 172 (Hf 172) Hafnium 181...

  19. Diagnosis and Treatment of Small Bowel Cancers Using Radioactive Gold Nanoparticles and Wireless Fluorescence Capsule Endoscopy

    PubMed Central

    Alizadeh, M.; Qaradaghi, V.

    2016-01-01

    Background Therapeutic and diagnosis properties of radioactive gold nanoparticle (198-AuNPs) cause them to be suitable for detection and treatment of tumors. Objective Electrical and optical properties of PEG-198AuNPs were examined in this paper. Polyethylene Glycol (PEG)-198 AuNPs can be used for treatment and diagnosis of small intestine tumors. Methods Wireless fluorescence capsule endoscopy will be able to detect emission lights of triggered Au by external light. First, the output electrical field was calculated by DDSCAT software. Secondly, tumor and distribution of PEG-198 gold nanoparticles were modeled using Monte Carlo simulation and finally dose delivered throughout a solid tumor when the PEG-198 gold nanoparticles linked to each cell was calculated. Results Polyethylene Glycol functionalized gold nanoparticles (AuNPs) possess optimized sizes (30 nm core diameter and 70 nm hydrodynamic diameters) to target individual tumor cells. Surface distribution to receive doses of up to 50Gy was simulated.  Activities and absorbed doses by the tumors with 0.25cm and 0.5cm radius were 187.9mCi and 300mCi and 72 and 118 Gy,respectively. Conclusion Therapeutic and diagnosis properties of 198-AuNPs show that it can be used for treatment and detection of small bowel tumors in early stage of growing. PMID:27026950

  20. Experience with fast neutron therapy for unresectable carcinoma of the pancreas

    SciTech Connect

    Al-Abdulla, A.S.M.; Hussey, D.H.; Olson, M.H.; Wright, A.E.

    1981-02-01

    The records of 70 patients with adenocarcinoma of the pancreas treated with radiotherapy were reviewed. Fifteen were treated with 50-MeV/sub d ..-->.. Be/ neutrons or a combination of 50-MeV/sub d ..-->.. Be/ neutrons and 25 to 32 MeV photons (neutron group), 30 with external beam photons alone (photon group), and 25 with radioactive gold-grain implantation (/sup 198/Au-implant group). The 12-month survival rate was 40% (6/15) for the neutron group; three patients in this group were living at the time of analysis, 16, 19, and 42 months from the date of diagnosis. By comparison, the 12-month survival rate was 23% (7/30) for the photon group and 32% (8/25) or the /sup 198/Au-implant group. The actuarial survival curve for the neutron group was significantly better than the survival curve for the photon group (Wilcoxon test/sup 7/: p = 0.3). Although the difference between the survival curves for the neutron and /sup 198/Au-implant groups is not statistically significant, the neutron patients presented more advanced disease than those treated with radioactive gold-grain implants. No radiotherapy complications were observed in the neutron group, whereas 3% (1/30) of patients in the photon group and 24% (6/25) of patients in the /sup 198/Au-implant group developed major complications.

  1. Methyl mercury dynamics in littoral sediments of a temperate seepage lake

    USGS Publications Warehouse

    Krabbenhoft, D.P.; Gilmour, C.C.; Benoit, J.M.; Babiarz, C.L.; Andren, A.W.; Hurley, J.P.

    1998-01-01

    The sites and rates of methyl mercury (MeHg) production and transport in littoral zone sediments were investigated at Pallette Lake in northern Wisconsin. In littoral areas where groundwater inflow occurs, sulfate supply from groundwater creates profiles of electron acceptors (sulfate) and donors (methane, sulfide) that are reversed from those found in sediments whose sulfate supply is delivered from overlying water. The highest MeHg concentrations in porewaters and the maximal advective MeHg flux rates (4.5-61.7 ng??m-2??day-1) were observed in the spring, while highest bulk phase concentrations occur later in the summer. These estimated MeHg fluxes are greater than the mean areal production rates estimated previously for the water column and are similar to the atmospheric flux. Gross MeHg production was measured using the addition of 203Hg as a tracer to sediments. The depth at which maximal 203Hg methylation occurred coincided with the observed maximums m solid-phase and porewater MeHg concentrations. Because input, advection, and accumulation of MeHg in these sediments were measured directly, an independent estimate of MeHg production could be made and compared with 203Hg-derived rates. This comparison suggests that the 203Hg tracer method provides reasonable estimates of gross methylation rates and that a substantial fraction of solid-phase Hg is available for methylation.

  2. Biokinetics of Hg and Pb accumulation in the encapsulated egg of the common cuttlefish Sepia officinalis: radiotracer experiments.

    PubMed

    Lacoue-Labarthe, T; Warnau, M; Metian, M; Oberhänsli, F; Rouleau, C; Bustamante, P

    2009-12-01

    Uptake and depuration kinetics of dissolved (203)Hg and (210)Pb were determined during the entire embryonic development of the eggs of the cuttlefish, Sepia officinalis (50d at 17 degrees C). (203)Hg and (210)Pb were accumulated continuously by the eggs all along the development time reaching load/concentration ratio (LCR) of 467+/-43 and 1301+/-126g, respectively. During the first month, most of the (203)Hg and (210)Pb remained associated with the eggshell indicating that the latter acted as an efficient shield against metal penetration. From this time onwards, (203)Hg accumulated in the embryo, indicating that it passed through the eggshell, whereas (210)Pb did not cross the chorion during the whole exposure time. It also demonstrated that translocation of Hg associated with the inner layers of the eggshell is a significant source of exposure for the embryo. This study highlighted that the maturing embryo could be subjected to the toxic effects of Hg in the coastal waters where the embryonic development is taking place.

  3. Bioavailability of trace contaminants ({sup 241}Am, {sup 57}Co, {sup 137}Cs) to a benthic bivalve from pore waters and sediments

    SciTech Connect

    Gagnon, C.; Stupakoff, I.; Fisher, N.S.

    1995-12-31

    Sediments are major repositories of contaminants in marine ecosystems and can serve as a source of some contaminants for benthic organisms. The authors used the clam Macoma balthica, a species employed in monitoring coastal contamination, to compare experimentally three uptake sources: overlying water, ingested surface sediment and anoxic pore water. They studied the bioavailability of selected radionuclides ({sup 241}Am, {sup 57}Co, {sup 137}Cs) representing a large range of particle reactivity. For comparison, the authors also used CH{sub 3} {sup 203}Hg, which is highly assimilated by marine organisms. Clams were exposed separately to contaminated overlying water, surface oxic sediment and anoxic sediment. Radioactivity in animals was determined at the end of the exposure period. {sup 137}CS, which is not particle reactive in seawater, was not bioaccumulated from any source. {sup 241}Am and {sup 57}Co concentration factors in clams obtained from overlying water were approximately an order of magnitude lower than that of CH{sub 3} {sup 203}Hg. Ingested oxidized sediment particles do not appear to be a significant source for these radionuclides. {sup 241}Am, {sup 57}Co and CH{sub 3} {sup 203}Hg were bioconcentrated from anoxic pore waters, but the highly particle-reactive {sup 241}Am was mostly adsorbed onto the clam`s shell. The bioconcentration of CH{sub 3} {sup 203}Hg from pore waters was, however, only one tenth of that from overlying water.

  4. Mercury methylation in mesocosms with and without the aquatic macrophyte Eichhornia crassipes (mart.) Solms.

    PubMed

    Correia, Raquel Rose Silva; Martins de Oliveira, Diana Ciannella; Guimarães, Jean Remy Davée

    2013-10-01

    Mercury is a toxic pollutant and spreads to several compartments in the environment. Previous in-vitro studies showed that roots of aquatic macrophytes are sites of methylmercury formation, performed mainly by sulfate-reducing bacteria (SRB). The objective of this study was to observe MMHg formation and distribution among filtered water (0.2µm), suspended and settled particles and macrophyte roots during seventeen days, in (203)Hg- spiked mesocosms with and without live Eichhornia crassipes whole plants and a SRB inhibitor. Root samples were also incubated in-vitro for comparison of MM(203)Hg formation under in-vitro and in-vivo conditions. To evaluate the effect of SRB inhibition by sodium molybdate on total heterotrophic activity, the latter was measured by (3)H-leucine uptake. Inhibition of Hg methylation by sodium molybdate decreased with time in mesocosms. MMHg averaged 10, 12.4 and 0.23 percent of total (203)Hg present in filtered water, suspended particles and roots respectively. In vitro MMHg formation in roots averaged 5.54 percent of total added (203)Hg, with a clearer SRB inhibition effect than in mesocosms. Though significant, MMHg formation in roots from in-vivo mesocosms was one order of magnitude lower than previously found in in-vitro incubations of roots alone.

  5. Mercury methylation by fish intestinal contents.

    PubMed Central

    Rudd, J W; Furutani, A; Turner, M A

    1980-01-01

    A new radiochemical method has been applied to the examination of mercury methylation in fish intestinal contents. Intestinal contents of six freshwater fish species were found capable of converting 203Hg2+ to CH3203Hg+. This activity was observed in fish from five of six lakes tested whether or not there was mercury pollution. Bacterial activity in the intestinal contents is most likely responsible for this methylation. Methylating activity of piscivors increased with decreasing quantity of intestinal contents. Generally, pike and walleye intestinal contents methylated a larger fraction of 203Hg2+ than those of whitefish and suckers. These data contradict the previous general conclusion that there is no mercury methylation in fish. PMID:7425625

  6. Gum arabic-coated radioactive gold nanoparticles cause no short-term local or systemic toxicity in the clinically relevant canine model of prostate cancer

    PubMed Central

    Axiak-Bechtel, Sandra M; Upendran, Anandhi; Lattimer, Jimmy C; Kelsey, James; Cutler, Cathy S; Selting, Kim A; Bryan, Jeffrey N; Henry, Carolyn J; Boote, Evan; Tate, Deborah J; Bryan, Margaret E; Katti, Kattesh V; Kannan, Raghuraman

    2014-01-01

    Introduction Gum arabic-coated radioactive gold nanoparticles (GA-198AuNPs) offer several advantages over traditional brachytherapy in the treatment of prostate cancer, including homogenous dose distribution and higher dose-rate irradiation. Our objective was to determine the short-term safety profile of GA-198AuNPs injected intralesionally. We proposed that a single treatment of GA-198AuNPs would be safe with minimal-to-no evidence of systemic or local toxicity. Methods Nine dogs with spontaneously occurring prostatic cancer were treated. Injections were performed with ultrasound or computerized tomography guidance. Complete blood counts, chemistry panels, and urinalyses were performed at weekly intervals for 1 month and imaging was repeated 4 weeks postinjection. Planar scintigraphic images were obtained within 30 minutes of injection. Results No statistically significant difference was found in any hematologic or biochemical parameter studied, nor was any evidence of tumor swelling or abscessation found in eight dogs with repeat imaging; one dog died secondary to urethral obstruction 12 days following injection. At 30 minutes postinjection, an average of 53% of injected dose in seven dogs was retained in the prostate, with loss of remaining activity in the bladder and urethra; no systemic uptake was detected. Conclusion GA-198AuNP therapy had no short-term toxicity in the treatment of prostatic cancer. While therapeutic agent was found in the prostate immediately following injection, some loss of agent was detected in the bladder and urethra. Localization of radioactivity within the prostate was lower than anticipated and likely due to normal vestigial prostatic ducts. Therefore, further study of retention, dosimetry, long-term toxicity, and efficacy of this treatment is warranted prior to Phase I trials in men. PMID:25378926

  7. Radiochemical separation of gold by amalgam exchange

    USGS Publications Warehouse

    Ruch, R.R.

    1970-01-01

    A rapid and simple method for the radiochemical separation of gold after neutron activation. The technique is based on treatment with a dilute indium-gold amalgam, both chemical reduction and isotopic exchange being involved. The counting efficiency for 198Au in small volumes of the amalgam is good. Few interferences occur and the method is applicable to clays, rocks, salts and metals. The possibility of determining silver, platinum and palladium by a similar method is mentioned. ?? 1970.

  8. Gum arabic-coated radioactive gold nanoparticles cause no short-term local or systemic toxicity in the clinically relevant canine model of prostate cancer.

    PubMed

    Axiak-Bechtel, Sandra M; Upendran, Anandhi; Lattimer, Jimmy C; Kelsey, James; Cutler, Cathy S; Selting, Kim A; Bryan, Jeffrey N; Henry, Carolyn J; Boote, Evan; Tate, Deborah J; Bryan, Margaret E; Katti, Kattesh V; Kannan, Raghuraman

    2014-01-01

    Gum arabic-coated radioactive gold nanoparticles (GA-(198)AuNPs) offer several advantages over traditional brachytherapy in the treatment of prostate cancer, including homogenous dose distribution and higher dose-rate irradiation. Our objective was to determine the short-term safety profile of GA-(198)AuNPs injected intralesionally. We proposed that a single treatment of GA-(198)AuNPs would be safe with minimal-to-no evidence of systemic or local toxicity. Nine dogs with spontaneously occurring prostatic cancer were treated. Injections were performed with ultrasound or computerized tomography guidance. Complete blood counts, chemistry panels, and urinalyses were performed at weekly intervals for 1 month and imaging was repeated 4 weeks postinjection. Planar scintigraphic images were obtained within 30 minutes of injection. No statistically significant difference was found in any hematologic or biochemical parameter studied, nor was any evidence of tumor swelling or abscessation found in eight dogs with repeat imaging; one dog died secondary to urethral obstruction 12 days following injection. At 30 minutes postinjection, an average of 53% of injected dose in seven dogs was retained in the prostate, with loss of remaining activity in the bladder and urethra; no systemic uptake was detected. GA-(198)AuNP therapy had no short-term toxicity in the treatment of prostatic cancer. While therapeutic agent was found in the prostate immediately following injection, some loss of agent was detected in the bladder and urethra. Localization of radioactivity within the prostate was lower than anticipated and likely due to normal vestigial prostatic ducts. Therefore, further study of retention, dosimetry, long-term toxicity, and efficacy of this treatment is warranted prior to Phase I trials in men.

  9. The Experimental and Calculated Total γ-RAY Spectra and Population of Levels up to 3 Mev of Heavy Nuclei

    NASA Astrophysics Data System (ADS)

    Khitrov, V. A.; Sukhovoj, A. M.; Grigoriev, E. P.

    2003-06-01

    Comparison between the experimental and calculated with the parameters [1] spectra demonstrates more or less improvement of their correspondence (up to practical coincidence) for the 114Cd, 158Gd, 166Ho, 168Er, 182Ta, 196Pt, 198Au, and 200Hg compound nuclei and some exceeding of the calculated intensity for 150Sm, 156Gd, 160Tb, 164Dy, 176Lu, and 192Ir at the γ-transition energy Eγ > 3 - 4 MeV.

  10. Permanent and removable implants for the brachytherapy of brain tumors

    SciTech Connect

    Gutin, P.H.; Phillips, T.L.; Hosobuchi, Y.

    1981-10-01

    Thirty-seven patients harboring primary or metastatic brain tumors were treated with 40 implantations of radioactive sources (/sup 192/Ir, /sup 198/Au, or /sup 125/I) using stereotactic neurosurgical techniques. Most tumors had recurred after surgery, whole brain irradiation, and treatment with all feasible chemotherapeutic agents. Sixteen of the 40 implants were pregnant; 24 were mounted in plastic catheters for removal after the desired dose had been delivered. One or more sources were placed in each tumor to deliver 3500-7350 rad to the tumor's periphery for /sup 198/Au, 4,000-12,000 rad for /sup 192/Ir, and 3,000-20,000 rad for /sup 125/I. Three of the six patients treated with /sup 192/Ir had objective responses for 2, 4, and 12 months, and two stabilized for 8 and 11 months. Seven of the 11 patients treated with /sup 198/Au were evaluable: three responded for 3, 5, and 37 + months, one deteriorating patient with a recurrent tumor stabilized for 6 months, and two deteriorated despite treatment. One patient received an interstitial ''boost'' dose with /sup 198/Au after whole brain irradiation and stabilized for 15 months before developing spinal metastases. Six patients received permanent implants with low activity /sup 125/I. Three of these patients had blioblastomas or anaplastic astrocytomas; all continued to deteriorate despite the interstitial irradiation, presumably because the dose rat was too low. One patient with a low-grade astrocytoma (optic chiasm) responded dramatically to permanent, low activity /sup 125/I implants (11 + months). Another (hypothalamic glioma) had a permanent /sup 125/I implant, responded, as was stable at 9 months when external irradiation was administered. One patient with a suprasellar ''teratoid'' tumor stabilized for 10 months.

  11. Control of mercury pollution.

    PubMed

    Noyes, O R; Hamdy, M K; Muse, L A

    1976-01-01

    When a 203Ng(NO3)2 solution was kept at 25 degrees C in glass or polypropylene containers, 50 and 80% of original radioactivity was adsorbed to the containers' walls after 1 and 4 days, respectively. However, no loss in radioactivity was observed if the solution was supplemented with HgCl as carrier (100 mug Hg2+/ml) and stored in either container for 13 days. When 203Hg2+ was dissolved in glucose basal salt broth with added carrier, levels of 203Hg2+ in solution (kept in glass) decreased to 80 and 70% of original after 1 and 5 days and decreased even more if stored in polypropylene (60 and 40% of original activity after 1 and 4 days, respectively). In the absence of carrier, decreases of 203Hg2+ activities in media stored in either container were more pronounced due to chemisorption (but) not diffusion. The following factors affecting the removal of mercurials from aqueous solution stored in glass were examined: type and concentration of adsorbent (fiber glass and rubber powder); pH; pretreatment of the rubber; and the form of mercury used. Rubber was equally effective in the adsorption of organic and inorganic mercury. The pH of the aqueous 203Hg2+ solution was not a critical factor in the rate of adsorption of mercury by the rubber. In addition, the effect of soaking the rubber in water for 18 hr did not show any statistical difference when compared with nontreated rubber. It can be concluded that rubber is a very effective adsorbent of mercury and, thus, can be used as a simple method for control of mercury pollution.

  12. In vivo integrity of polymer-coated gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Kreyling, Wolfgang G.; Abdelmonem, Abuelmagd M.; Ali, Zulqurnain; Alves, Frauke; Geiser, Marianne; Haberl, Nadine; Hartmann, Raimo; Hirn, Stephanie; de Aberasturi, Dorleta Jimenez; Kantner, Karsten; Khadem-Saba, Gülnaz; Montenegro, Jose-Maria; Rejman, Joanna; Rojo, Teofilo; de Larramendi, Idoia Ruiz; Ufartes, Roser; Wenk, Alexander; Parak, Wolfgang J.

    2015-07-01

    Inorganic nanoparticles are frequently engineered with an organic surface coating to improve their physicochemical properties, and it is well known that their colloidal properties may change upon internalization by cells. While the stability of such nanoparticles is typically assayed in simple in vitro tests, their stability in a mammalian organism remains unknown. Here, we show that firmly grafted polymer shells around gold nanoparticles may degrade when injected into rats. We synthesized monodisperse radioactively labelled gold nanoparticles (198Au) and engineered an 111In-labelled polymer shell around them. Upon intravenous injection into rats, quantitative biodistribution analyses performed independently for 198Au and 111In showed partial removal of the polymer shell in vivo. While 198Au accumulates mostly in the liver, part of the 111In shows a non-particulate biodistribution similar to intravenous injection of chelated 111In. Further in vitro studies suggest that degradation of the polymer shell is caused by proteolytic enzymes in the liver. Our results show that even nanoparticles with high colloidal stability can change their physicochemical properties in vivo.

  13. In vivo integrity of polymer-coated gold nanoparticles.

    PubMed

    Kreyling, Wolfgang G; Abdelmonem, Abuelmagd M; Ali, Zulqurnain; Alves, Frauke; Geiser, Marianne; Haberl, Nadine; Hartmann, Raimo; Hirn, Stephanie; de Aberasturi, Dorleta Jimenez; Kantner, Karsten; Khadem-Saba, Gülnaz; Montenegro, Jose-Maria; Rejman, Joanna; Rojo, Teofilo; de Larramendi, Idoia Ruiz; Ufartes, Roser; Wenk, Alexander; Parak, Wolfgang J

    2015-07-01

    Inorganic nanoparticles are frequently engineered with an organic surface coating to improve their physicochemical properties, and it is well known that their colloidal properties may change upon internalization by cells. While the stability of such nanoparticles is typically assayed in simple in vitro tests, their stability in a mammalian organism remains unknown. Here, we show that firmly grafted polymer shells around gold nanoparticles may degrade when injected into rats. We synthesized monodisperse radioactively labelled gold nanoparticles ((198)Au) and engineered an (111)In-labelled polymer shell around them. Upon intravenous injection into rats, quantitative biodistribution analyses performed independently for (198)Au and (111)In showed partial removal of the polymer shell in vivo. While (198)Au accumulates mostly in the liver, part of the (111)In shows a non-particulate biodistribution similar to intravenous injection of chelated (111)In. Further in vitro studies suggest that degradation of the polymer shell is caused by proteolytic enzymes in the liver. Our results show that even nanoparticles with high colloidal stability can change their physicochemical properties in vivo.

  14. Radioluminescent gold nanocages with controlled radioactivity for real-time in vivo imaging.

    PubMed

    Wang, Yucai; Liu, Yongjian; Luehmann, Hannah; Xia, Xiaohu; Wan, Dehui; Cutler, Cathy; Xia, Younan

    2013-02-13

    Cerenkov luminescence imaging based on light emission from the decay of radionuclides has recently drawn great interest in molecular imaging. In this paper, we report for the first time the Cerenkov luminescence phenomenon of (198)Au isotope, as well as a facile route to the preparation of radioluminescent Au nanocages without additional radiolabeling or dye conjugation. The specific radioactivity of the Au nanocages could be easily and precisely controlled by varying the concentration of H(198)AuCl(4) precursor used for the galvanic replacement reaction. The direct incorporation of (198)Au atoms into the structure of Au nanocages enabled the ability of accurate analysis and real-time imaging in vivo. Furthermore, under biological conditions the radioactive Au nanocages were shown to emit light with wavelengths in the visible and near-infrared regions, enabling luminescence imaging of the whole mice in vivo, as well as the organs ex vivo. When combined with their favorable scattering and absorption properties in the near-infrared region, the radioactive Au nanocages can serve as a new platform for multimodality imaging and will have a significant impact on both small animal and clinical imaging.

  15. Mercury and flooding cycles in the Tapajós River basin, Brazilian Amazon: the role of periphyton of a floating macrophyte (Paspalum repens).

    PubMed

    Coelho-Souza, Sérgio A; Guimarães, Jean R D; Miranda, Márcio R; Poirier, Hugo; Mauro, Jane B N; Lucotte, Marc; Mergler, Donna

    2011-06-15

    Methylmercury (MeHg) increases mercury (Hg) toxicity and is biomagnified in the trophic chain contaminating riverine Amazon populations. Freshwater macrophyte roots are a main site of Hg methylation in different Brazilian environments. Paspalum repens periphyton was sampled in four floodplain lakes during the dry, rainy and wet seasons for measurement of total Hg (THg), MeHg, Hg methylation potentials, %C, %N, δ(13)C, δ(15)N and bacterial heterotrophic production as (3)H-leucine incorporation rate. THg concentration varied from 67 to 198 ng/g and the potential of Me(203)Hg formation was expressive (1-23%) showing that periphyton is an important matrix both in the accumulation of Hg and in MeHg production. The concentration of MeHg varied from 1 to 6 ng/g DW and was positively correlated with Me(203)Hg formation. Though methylmercury formation is mainly a bacterial process, no significant correlation was observed between the methylation potentials and bacterial production. The multiple regressions analyses suggested a negative correlation between THg and %C and %N and between methylation potential and δ(13)C. The discriminant analysis showed a significant difference in periphyton δ(15)N, δ(13)C and THg between seasons, where the rainy season presented higher δ(15)N and the wet period lighter δ(13)C, lower THg values and higher Me(203)Hg formation. This exploratory study indicates that the flooding cycle could influence the periphyton composition, mercury accumulation and methylmercury production.

  16. Laboratory Study of Chemical Speciation of Mercury in Lake Sediment and Water under Aerobic and Anaerobic Conditions

    PubMed Central

    Regnell, Olof; Tunlid, Anders

    1991-01-01

    Chemical speciation and partitioning of radiolabeled HgCl2 were studied in model aquatic systems consisting of undisturbed eutrophic lake sediment and water in plastic cylinders. The cylinders were either gradually made anaerobic by a gentle flow of N2-CO2 or kept aerobic by air flow. The proportion of methylated 203Hg was significantly higher, in both water and sediment, in the anaerobic systems than in the aerobic systems. The composition and total concentration of fatty acids originating from bacterial phospholipids, as well as the concentration of vitamin B12, including related cobalamins, were similar in sediments from the anaerobic and aerobic systems. Bacterial cell numbers were, on average, 3.6 times higher in the anaerobic water columns than in the aerobic ones. Volatilization of 203Hg occurred in all systems except in an autoclaved control and was of similar magnitudes in the anaerobic and aerobic systems. Incorporation of 203Hg into the sediment was significantly faster in the aerobic systems than in the anaerobic systems. These results suggest that episodes of anoxia in bottom waters and sediment cause an increase in net mercury methylation and, hence, an increase in bioavailable mercury. PMID:16348444

  17. Low-Dose Prostate Cancer Brachytherapy with Radioactive Palladium-Gold Nanoparticles.

    PubMed

    Laprise-Pelletier, Myriam; Lagueux, Jean; Côté, Marie-France; LaGrange, Thomas; Fortin, Marc-André

    2017-02-01

    Prostate cancer (PCa) is one of the leading causes of death among men. Low-dose brachytherapy is an increasingly used treatment for PCa, which requires the implantation of tens of radioactive seeds. This treatment causes discomfort; these implants cannot be removed, and they generate image artifacts. In this study, the authors report on intratumoral injections of radioactive gold nanoparticles (Au NPs) as an alternative to seeds. The particles ((103) Pd:Pd@Au-PEG and (103) Pd:Pd@(198) Au:Au-PEG; 10-14 nm Pd@Au core, 36-48 nm hydrodynamic diameter) are synthesized by a one-pot process and characterized by electron microscopy. Administrated as low volume (2-4 µL) single doses (1.6-1.7 mCi), the particles are strongly retained in PCa xenograft tumors, impacting on their growth rate. After 4 weeks, a tumor volume inhibition of 56% and of 75%, compared to the controls, is observed for (103) Pd:Pd@Au-PEG NPs and (103) Pd:Pd@(198) Au:Au-PEG NPs, respectively. Skin necrosis is observed with (198) Au; therefore, Au NPs labeled with (103) Pd only are a more advisable choice. Overall, this is the first study confirming the impact of (103) Pd@Au NPs on tumor growth. This new brachytherapy procedure could allow tunable doses of radioactivity, administered with smaller needles than with the current technologies, and leading to fewer image artifacts. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Measurement of thermal neutron cross-section and resonance integral for the 165Ho(n,γ) 166gHo reaction using electron linac-based neutron source

    NASA Astrophysics Data System (ADS)

    Nguyen, Van Do; Pham, Duc Khue; Kim, Tien Thanh; Kim, Guinyun; Lee, Manwoo; Kim, Kyung Sook; Kang, Heung-Sik; Cho, Moo-Hyun; Ko, In Soo; Namkung, Won

    2011-01-01

    The thermal neutron cross-section and the resonance integral of the 165Ho(n,γ) 166gHo reaction have been measured by the activation method using a 197Au(n,γ) 198Au monitor reaction as a single comparator. The high-purity natural Ho and Au foils with and without a cadmium shield case of 0.5 mm thickness were irradiated in a neutron field of the Pohang neutron facility. The induced activities in the activated foils were measured with a calibrated p-type high-purity Ge detector. The correction factors for the γ-ray attenuation ( Fg), the thermal neutron self-shielding ( Gth), the resonance neutron self-shielding ( Gepi) effects, and the epithermal neutron spectrum shape factor ( α) were taken into account. The thermal neutron cross-section for the 165Ho(n,γ) 166gHo reaction has been determined to be 59.7 ± 2.5 barn, relative to the reference value of 98.65 ± 0.09 barn for the 197Au(n,γ) 198Au reaction. By assuming the cadmium cut-off energy of 0.55 eV, the resonance integral for the 165Ho(n,γ) 166gHo reaction is 671 ± 47 barn, which is determined relative to the reference value of 1550 ± 28 barn for the 197Au(n,γ) 198Au reaction. The present results are, in general, good agreement with most of the previously reported data within uncertainty limits.

  19. Bioaccumulation of radionuclides in fertilized Canadian Shield lake basins.

    PubMed

    Bird, G A; Hesslein, R H; Mills, K H; Schwartz, W J; Turner, M A

    1998-07-11

    Radionuclide tracers of heavy metals (59Fe, 60Co, 65Zn, 75Se, 85Sr, 134Cs and 203Hg) representing potential contamination from nuclear power plants, industry and agriculture were added to separate basins of Lake 226, Experimental Lakes Area, northwestern Ontario. The two basins were part of a eutrophication experiment and differed in their trophic status; the north basin (L226N) was eutrophic whereas the south basin (L226S) was mesotrophic. Our objective was to determine the uptake of the radionuclides by biota and the effect of lake trophic status on their bioaccumulation. The trophic status of the lakes did not appear to have a marked effect on the accumulation of radionuclides by the biota. This may have been because of a mid-summer leakage of nutrients between the basins which enhanced primary production in L226S, because there is a time lag between primary production and the availability of the radionuclides to the fishes or because trophic status does not affect the uptake of at least some of these radionuclides. However, there was a tendency for faster uptake of the radionuclides in L226N by fish than L226S, but the differences were not significant. Concentrations in the biota generally decreased in the order: fathead minnow > pearl dace > tadpoles > slimy sculpin > leeches. Concentrations in biota generally decreased in the order. 65Zn > 203Hg > 75Se > 134Cs > 60Co > 85Sr = 59Fe. Cobalt-60 concentrations in tadpoles were greater than in the other biota. Radionuclide concentrations in the tissues of lake whitefish indicated that uptake was predominantly from food. Radionuclide concentrations were usually higher in the posterior gut, liver and kidney than in other tissues, whereas body burdens were generally high in the muscle for 75Se, 134Cs and 203Hg; kidney and gut for 60Co; and bone for 65Zn and 75Se. Mercury-203 burdens were also high in the bone and gut.

  20. Mercury Distribution, Methylation and Volatilization in Microcosms with and without the Sea Anemone Bunodosoma caissarum

    NASA Astrophysics Data System (ADS)

    Ansari, N. R.; Correia, R. R. S.; Fernandez, M. A. S.; Cordeiro, R. C.; Guimarães, J. R. D.

    2014-12-01

    Mercury (Hg) can be a dangerous contaminant and has a complex biogeochemical cycling in aquatic environments. The sea anemone Bunodosoma caissarum is an endemic species in Brazil capable of bioaccumulating Hg from the ambient seawater. The radiotracer 203Hg was used in order to investigate mechanisms of Hg uptake and depuration of B. caissarum and the distribution of Hg in laboratory model systems, with and without B. caissarum. A single initial spike of 203Hg was added to each microcosm. Microcosms had continuous air renovation and trapping of Hg volatile forms. Total Hg in different compartments was measured by gamma spectrometry. In the uptake experiment 203Hg activity was determined periodically in seawater and specimens for 6 days. At the end, specimens had an average bioconcentration factor of 70. After the uptake experiment, methylmercury (MeHg) in seawater was extracted and measured by liquid scintillation. In microcosms with and without B. caissarum, respectively 0.05% and 0.32% of the initial spike was found as MeHg. Hg was probably less available for methylation in the first because of bioaccumulation and higher concentrations of suspended particulate matter that could form complexes with Hg. After that, specimens were transferred to unspiked microcosms. After a 48 day depuration specimens still retained 35 - 70% of the previously bioaccumulated Hg and 0.2 - 2.4% of the total Hg was MeHg. The presence of B. caissarum resulted in an unexpected higher volatilization of Hg (58%) compared to controls (17%). This increased volatilization is possibly a result of Hg2+ reduction mediated by microorganisms associated with its tissues and mucus secretions and/or an unknown defense mechanism of this species.

  1. Biliary-hepatic recycling of a xenobiotic: gallbladder absorption of methyl mercury.

    PubMed

    Dutczak, W J; Clarkson, T W; Ballatori, N

    1991-06-01

    The role of the gallbladder in the disposition of methyl mercury was investigated in guinea pig, hamster, and macaque monkey. 203Hg-labeled methyl mercury or inorganic mercury (5 microM) and [14C]inulin were instilled into the in situ guinea pig or hamster gallbladder. After 2 h, only 27.6 +/- 7.0% of the methyl mercury remained in guinea pig gallbladder fluid as compared with 85.0 +/- 3.2% of the inorganic mercury and 90.7 +/- 4.5% of the [14C]-inulin. In the hamster, 42.5 +/- 4.5% of methyl mercury and 95% +/- 0.9% of inorganic mercury remained after 2 h. When the sulfhydryl-containing compounds L-cysteine, glutathione, and bovine serum albumin (20 microM) were added to the test solution, cysteine increased and albumin decreased absorption of methyl mercury. Ligation of guinea pig cystic artery decreased gallbladder fluid absorption from 72.7 +/- 8.6 to 26.5 +/- 9.8% over 2 h but did not alter methyl mercury absorption. Bile was also sampled from gallbladders of four monkeys exposed chronically to CH3HgCl and from three control monkeys. For one of the exposed and one of the control monkeys, bile was also collected from the common hepatic duct. In both methyl mercury-exposed and control monkeys, the concentration of methyl mercury in gallbladder bile was lower than in hepatic bile. In contrast, the concentration of inorganic mercury in gallbladder bile was four to seven times that of hepatic bile, suggesting that methyl mercury but not inorganic mercury was being reabsorbed. To assess the functional significance of methyl mercury reabsorption by the gallbladder, guinea pig cystic ducts were ligated, the animals were given CH3 203HgCl (10 mumol/kg iv), and body burden of 203Hg was measured over 16 days.(ABSTRACT TRUNCATED AT 250 WORDS)

  2. Accelerated methylmercury elimination in gamma-glutamyl transpeptidase-deficient mice.

    PubMed Central

    Ballatori, N.; Wang, W.; Lieberman, M. W.

    1998-01-01

    The disposition and toxicity of methylmercury, a ubiquitous environmental pollutant, is modulated by binding to the endogenous tripeptide glutathione (GSH) and metabolism of the resulting methylmercury-glutathione complex by the ectoproteins gamma-glutamyl transpeptidase (GGT) and dipeptidase. To evaluate the role of GGT in the whole-body disposition of methylmercury, we compared the elimination of [203Hg]methylmercury in GGT-deficient mice with that in wild-type mice and mice heterozygous for this deficiency. The effects of N-acetylcysteine (NAC), a drug used to maintain the cysteine and GSH levels of GGT-deficient mice, were also examined. Female mice were treated with either 0.5 or 25 micromol of CH3 203HgCl/kg body weight, in the presence and absence of 10 mg/ml NAC in the drinking water. There were no differences in methylmercury excretion between the wild-type and heterozygous mice; however, the GGT-deficient mice excreted methylmercury more rapidly at both dose levels. Wild-type and heterozygous mice excreted from 11 to 24% of the dose in the first 48 hours, whereas the GGT-deficient mice excreted 55 to 66% of the dose, with most of the methylmercury being excreted in urine. Urinary methylmercury excretion was further accelerated in mice that received NAC. In contrast to methylmercury, the whole-body elimination of inorganic mercury was not affected by GGT deficiency, although the tissue distribution of inorganic mercury was markedly different in GGT-deficient male mice, with only 13% of the 203Hg body burden in the kidneys of GGT-deficient mice versus approximately 50% in kidneys of wild-type male mice. These findings provide direct evidence for a major role of GGT in regulating the tissue distribution and elimination of methylmercury and inorganic mercury and provide additional support for the use of NAC as an antidote in methylmercury poisoning. PMID:9546365

  3. Absolute measurement of activity of ¹⁹⁸Au foils using the 4πβ-γ coincidence counting method and corrections by the Monte Carlo simulation.

    PubMed

    Nishiyama, Jun; Harano, Hideki; Matsumoto, Tetsuro; Sato, Yasushi; Uritani, Akira; Kudo, Katsuhisa

    2012-01-01

    The thermal neutron fluence rate is determined by the gold activation method. The absolute activity of the irradiated gold foil is measured by a 4πβ-γ coincidence counter. Using this method, corrections for the detection of conversion electrons and gamma rays by a 4πβ counter are very important to obtain accurate absolute activity. In this work, Monte Carlo simulations were performed to derive the correction factor K. The absolute measurement of (198)Au activity for 20-100 μm thickness Au foils were performed to verify the calculating model of the 4πβ-γ coincidence counting system.

  4. Do radioactive half-lives vary with the Earth-to-Sun distance?

    PubMed

    Hardy, J C; Goodwin, J R; Iacob, V E

    2012-09-01

    Recently, Jenkins, Fischbach and collaborators have claimed evidence that radionuclide half-lives vary systematically over a ±0.1% range as a function of the oscillating distance between the Earth and the Sun, based on multi-year activity measurements. We have avoided the time-dependent instabilities to which such measurements are susceptible by directly measuring the half-life of (198)Au (t(1/2)=2.695 d) on seven occasions spread out in time to cover the complete range of Earth-Sun distances. We observe no systematic oscillations in half-life and can set an upper limit on their amplitude of ±0.02%.

  5. Thermal neutron cross-section and resonance integral of the 152Sm(n,γ)153Sm reaction induced by pulsed neutrons

    NASA Astrophysics Data System (ADS)

    Van Do, Nguyen; Khue, Pham Duc; Thanh, Kim Tien; Hien, Nguyen Thi; Kim, Guinyun; Kim, Kwangsoo; Shin, Sung-Gyun; Kye, Yong-Uk; Cho, Moo-Hyun

    2017-10-01

    We measured the thermal neutron cross-section (σ0) and resonance integral (I0) of the 152Sm(n,γ)153Sm reaction relative to that of the 197Au(n,γ)198Au reaction. Sm and Au foils with and without a cadmium cover of 0.5 mm were irradiated with moderated pulsed neutrons produced from the electron linac. The induced activities of the reaction products were determined via high energy resolution HPGe detector. The present results: σ0,Sm =212±8 b and I0,Sm =3.02±0.19 kb are consistent with most of the existing reference data.

  6. Neutron-capture gamma rays below 40 keV

    NASA Astrophysics Data System (ADS)

    Durner, P.; Von Egidy, T.; Hartmann, F. J.

    1989-06-01

    A review of neutron-capture gamma ray measurements below 40 keV is given and experimental methods are discussed. New experiments with a Si(Li) detector have been performed. Energies and absolute intensities of low energy (n, γ) transitions in 28Al, 40K, 52V, 128I, 134Cs, 160Tb, 166Ho, 170Tm, 176Lu, 182Ta, 192Ir, 198Au and 233Th are presented. These new results can serve calibration purposes and provide nuclear structure information.

  7. Report on First Activations with the Lead Slowing Down Spectrometer

    SciTech Connect

    Warren, Glen A.; Mace, Emily K.; Pratt, Sharon L.; Stave, Sean; Woodring, Mitchell L.

    2011-03-03

    On Feb. 17 and 18 2011, six items were irradiated with neutrons using the Lead Slowing Down Spectrometer. After irradiation, dose measurements and gamma-spectrometry measurements were completed on all of the samples. No contamination was found on the samples, and all but one provided no dose. Gamma-spectroscopy measurements qualitatively agreed with expectations based on the materials, with the exception of silver. We observed activation in the room in general, mostly due to 56Mn and 24Na. Most of the activation was short lived, with half-lives on the scale of hours, except for 198Au which has a half-life of 2.7 d.

  8. Effects of ocean acidification on trace element accumulation in the early-life stages of squid Loligo vulgaris.

    PubMed

    Lacoue-Labarthe, T; Réveillac, E; Oberhänsli, F; Teyssié, J L; Jeffree, R; Gattuso, J P

    2011-09-01

    The anthropogenic release of carbon dioxide (CO(2)) into the atmosphere leads to an increase in the CO(2) partial pressure (pCO(2)) in the ocean, which may reach 950 μatm by the end of the 21st century. The resulting hypercapnia (high pCO(2)) and decreasing pH ("ocean acidification") are expected to have appreciable effects on water-breathing organisms, especially on their early-life stages. For organisms like squid that lay their eggs in coastal areas where the embryo and then paralarva are also exposed to metal contamination, there is a need for information on how ocean acidification may influence trace element bioaccumulation during their development. In this study, we investigated the effects of enhanced levels of pCO(2) (380, 850 and 1500 μatm corresponding to pH(T) of 8.1, 7.85 and 7.60) on the accumulation of dissolved (110m)Ag, (109)Cd, (57)Co, (203)Hg, (54)Mn and (65)Zn radiotracers in the whole egg strand and in the different compartments of the egg of Loligo vulgaris during the embryonic development and also in hatchlings during their first days of paralarval life. Retention properties of the eggshell for (110m)Ag, (203)Hg and (65)Zn were affected by the pCO(2) treatments. In the embryo, increasing seawater pCO(2) enhanced the uptake of both (110m)Ag and (65)Zn while (203)Hg showed a minimum concentration factor (CF) at the intermediate pCO(2). (65)Zn incorporation in statoliths also increased with increasing pCO(2). Conversely, uptake of (109)Cd and (54)Mn in the embryo decreased as a function of increasing pCO(2). Only the accumulation of (57)Co in embryos was not affected by increasing pCO(2). In paralarvae, the CF of (110m)Ag increased with increasing pCO(2), whereas the (57)Co CF was reduced at the highest pCO(2) and (203)Hg showed a maximal uptake rate at the intermediate pCO(2). (54)Mn and (65)Zn accumulation in paralarvae were not significantly modified by hypercapnic conditions. Our results suggest a combined effect of pH on the adsorption and

  9. Proposed experimental test of Bell's inequality in nuclear beta decay

    SciTech Connect

    Skalsey, M.

    1986-04-15

    A ..beta.. decay experiment is proposed for testing Bell's inequality, related to hidden-variables alternatives to quantum mechanics. The experiment uses Mott scattering for spin polarization analysis of internal conversion electrons. Beta-decay electrons, in cascade with the conversion electrons, are longitudinally polarized due to parity violation in the weak interaction. So simply detecting the ..beta.. electron direction effectively measures the spin. A two-particle spin-spin correlation can thus be investigated and related, within certain assumptions, to Bell's inequality. The example of /sup 203/Hg decay is used for a calculation of expected results. Specific problems related to nuclear structure and experimental inconsistencies are also discussed.

  10. Comparative effects of N,N-disubstituted dithiocarbamates and dimercaptosuccinate on mobilization of methylmercury in mice

    SciTech Connect

    Gale, G.R.; Atkins, L.M.; Smith, A.B.; Walker, E.M. Jr.; Jones, M.M.

    1984-07-01

    Diethyldithiocarbamate (DDTC) and five of its N,N-disubstituted analogs were compared with dimercaptosuccinate (DMSA) for effectiveness in mobilizing and promoting excretion of methylmercury (MeHg) following administration of a sublethal dose of MeHg labeled with Me203Hg. The previously reported effectiveness of DMSA was confirmed. Of the dithiocarbamates assessed, only di(carboxymethyl)-dithiocarbamate (DCDC) was active in mobilizing MeHg, and its activity on a molar dose basis was less than that of DMSA. The disparate structural requirements of dithiocarbamates as antagonists of MeHg and of cadmium were discussed.

  11. Benchmark experiments for cyclotron-based neutron source for BNCT.

    PubMed

    Yonai, S; Itoga, T; Baba, M; Nakamura, T; Yokobori, H; Tahara, Y

    2004-11-01

    In the previous study, we found the feasibility of a cyclotron-based BNCT using the Ta(p,n) neutrons at 90 degrees bombarded by 50 MeV protons, and the iron, AlF(3), Al and (6)LiF moderators by simulations using the MCNPX code. In order to validate the simulations to realize the cyclotron-based BNCT, we measured the epithermal neutron energy spectrum passing through the moderators with our new spectrometer consisting of a (3)He gas counter covered with a silicon rubber loaded with (nat)B and polyethylene moderator and the depth distribution of the reaction rates of (197)Au(n,gamma)(198)Au in an acrylic phantom set behind the rear surface of the moderators. The measured results were compared with the calculations using the MCNPX code. We obtained the good agreement between the calculations and measurements within approximately 10% for the neutron energy spectra and within approximately 20% for the depth distribution of the reaction rates of (197)Au(n,gamma)(198)Au in the phantom. The comparison clarified a good accuracy of the calculation of the neutron energy spectrum passing through the moderator and the thermalization in a phantom. These experimental results will be a good benchmark data to evaluate the accuracy of the calculation code.

  12. Dual radiolabeling as a technique to track nanocarriers: the case of gold nanoparticles.

    PubMed

    Rambanapasi, Clinton; Barnard, Nicola; Grobler, Anne; Buntting, Hylton; Sonopo, Molahlehi; Jansen, David; Jordaan, Anine; Steyn, Hendrik; Zeevaart, Jan Rijn

    2015-07-16

    Gold nanoparticles (AuNPs) have shown great potential for use in nanomedicine and nanotechnologies due to their ease of synthesis and functionalization. However, their apparent biocompatibility and biodistribution is still a matter of intense debate due to the lack of clear safety data. To investigate the biodistribution of AuNPs, monodisperse 14-nm dual-radiolabeled [14C]citrate-coated [198Au]AuNPs were synthesized and their physico-chemical characteristics compared to those of non-radiolabeled AuNPs synthesized by the same method. The dual-radiolabeled AuNPs were administered to rats by oral or intravenous routes. After 24 h, the amounts of Au core and citrate surface coating were quantified using gamma spectroscopy for 198Au and liquid scintillation for the 14C. The Au core and citrate surface coating had different biodistribution profiles in the organs/tissues analyzed, and no oral absorption was observed. We conclude that the different components of the AuNPs system, in this case the Au core and citrate surface coating, did not remain intact, resulting in the different distribution profiles observed. A better understanding of the biodistribution profiles of other surface attachments or cargo of AuNPs in relation to the Au core is required to successfully use AuNPs as drug delivery vehicles.

  13. Cerebral aneurysms following radiotherapy for medulloblastoma

    SciTech Connect

    Benson, P.J.; Sung, J.H.

    1989-04-01

    Three patients, two males and one female aged 21, 14, and 31 years, respectively, developed cerebral saccular aneurysms several years after undergoing radiotherapy for cerebellar medulloblastoma at 2, 5, and 14 years of age, respectively. Following surgery, all three received combined cobalt-60 irradiation and intrathecal colloidal radioactive gold (/sup 198/Au) therapy, and died from rupture of the aneurysm 19, 9, and 17 years after the radiotherapy, respectively. Autopsy examination revealed no recurrence of the medulloblastoma, but widespread radiation-induced vasculopathy was found at the base of the brain and in the spinal cord, and saccular aneurysms arose from the posterior cerebral arteries at the basal cistern or choroidal fissure. The aneurysms differed from the ordinary saccular aneurysms of congenital type in their location and histological features. Their locations corresponded to the areas where intrathecally administered colloidal /sup 198/Au is likely to pool, and they originated directly from a segment of the artery rather than from a branching site as in congenital saccular aneurysms. It is, therefore, concluded that the aneurysms in these three patients were most likely radiation-induced.

  14. A mercury saturation assay for measuring metallothionein in fish

    SciTech Connect

    Dutton, M.D. . Dept. of Zoology); Stephenson, M. . Environmental Science Branch); Klaverkamp, J.F. )

    1993-07-01

    An accurate, rapid, sensitive, and simple method using mercury saturation for quantifying metallothionein (MT) is described. A complex solution of enzymatic and nonenzymatic thiols, including rabbit liver MT-2, and supernatants from homogenized samples of rainbow trout liver were incubated in the presence of [sup 203]Hg in 10% trichloroacetic acid. Excess Hg was bound to an removed by chicken egg albumin, which denatured on contact with the acidic assay medium. After centrifugation, MT labeled with [sup 203]Hg remained in the TCA supernatant and was estimated using known stoichiometry for Hg-MT binding. A dilution series was used to establish that nonspecific metal binding, a common problem with other metal saturation assays, is negligible. Analysis of hepatic MT with high Cu content from rainbow trout demonstrated virtually complete displacement of Cu, Cd, and Zn by Hg. When compared to other metal-saturation assays developed for vertebrates, this method requires the least number of technical steps, and one-third or less of total preparatory and analytical time.

  15. Constitutive synthesis of a transport function encoded by the Thiobacillus ferrooxidans merC gene cloned in Escherichia coli

    SciTech Connect

    Kusano, Tomonobu Akita Prefectural College of Agriculture ); Ji, Guangyong; Silver, S. ); Inoue, Chihiro )

    1990-05-01

    Mercuric reductase activity determined by the Thiobacillus ferrooxidans merA gene (cloned and expressed constitutively in Escherichia coli) was measured by volatilization of {sup 203}Hg{sup 2+}. (The absence of a merR regulatory gene in the cloned Thiobacillus mer determinant provides a basis for the constitutive synthesis of this system.) In the absence of the Thiobacillus merC transport gene, the mercury volatilization activity was cryptic and was not seen with whole cells but only with sonication-disrupted cells. The Thiobacillus merC transport function was compared with transport via the merT-merP system of plasmid pDU1358. Both systems, cloned and expressed in E. coli, governed enhanced uptake of {sup 203}Hg{sup 2+} in a temperature- and concentration-dependent fashion. Uptake via MerT-MerP was greater and conferred greater hypersensitivity to Hg{sup 2+} than did uptake with MerC. Mercury uptake was inhibited by N-ethylmaleimide but not by EDTA. Ag{sup +} salts inhibited mercury uptake by the MerT-MerP system but did not inhibit uptake via MerC. Radioactive mercury accumulated by the MerT-MerP and by the MerC systems was exchangeable with nonradioactive Hg{sup 2+}.

  16. Mercury mass measurement in fluorescent lamps via neutron activation analysis

    NASA Astrophysics Data System (ADS)

    Viererbl, L.; Vinš, M.; Lahodová, Z.; Fuksa, A.; Kučera, J.; Koleška, M.; Voljanskij, A.

    2015-11-01

    Mercury is an essential component of fluorescent lamps. Not all fluorescent lamps are recycled, resulting in contamination of the environment with toxic mercury, making measurement of the mercury mass used in fluorescent lamps important. Mercury mass measurement of lamps via instrumental neutron activation analysis (NAA) was tested under various conditions in the LVR-15 research reactor. Fluorescent lamps were irradiated in different positions in vertical irradiation channels and a horizontal channel in neutron fields with total fluence rates from 3×108 cm-2 s-1 to 1014 cm-2 s-1. The 202Hg(n,γ)203Hg nuclear reaction was used for mercury mass evaluation. Activities of 203Hg and others induced radionuclides were measured via gamma spectrometry with an HPGe detector at various times after irradiation. Standards containing an Hg2Cl2 compound were used to determine mercury mass. Problems arise from the presence of elements with a large effective cross section in luminescent material (europium, antimony and gadolinium) and glass (boron). The paper describes optimization of the NAA procedure in the LVR-15 research reactor with particular attention to influence of neutron self-absorption in fluorescent lamps.

  17. Mercury methylation in sediments of a Brazilian mangrove under different vegetation covers and salinities.

    PubMed

    de Oliveira, Diana Ciannella Martins; Correia, Raquel Rose Silva; Marinho, Claudio Cardoso; Guimarães, Jean Remy Davée

    2015-05-01

    The presence and formation of methylmercury (MMHg), a highly toxic form of Hg, in mangrove ecosystems is poorly studied. Therefore the aim of this study was to evaluate mercury methylation potentials in sediment, litter and root samples (Avicennia shaueriana and Spartina alterniflora) from different regions of a mangrove ecosystem, as well as the influence of salinity on methylation. Sediment was sampled under different depths and in mangrove regions with different plant covers and salinities. All samples were incubated with (203)Hg and MM(203)Hg was extracted and measured by liquid scintillation. MMHg was formed in all samples and sites tested including plant roots and litter. Higher Hg methylation was found in the superficial fraction of sediments (0.47-7.82%). Infralittoral sandy sediment had low MMHg formation (0.44-1.61%). Sediment under Rhizophora mangle had lower MMHg formation (0.018-2.23%) than under A. shaueriana (0.2-4.63%) and Laguncularia racemosa (0.08-7.82). MMHg formation in sediment tended to increase with salinity but the differences were not significant. Therefore, MMHg formation occurs in different sites of mangrove ecosystems and may be an important threat that requires further study. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Luminescent gold nanoparticles for bioimaging

    NASA Astrophysics Data System (ADS)

    Zhou, Chen

    Inorganic nanoparticles (NPs) with tunable and diverse material properties hold great potential as contrast agents for better disease management. Over the past decades, luminescent gold nanoparticles (AuNPs) with intrinsic emissions ranging from the visible to the near infrared have been synthesized and emerge as a new class of fluorophores for bioimaging. This dissertation aims to fundamentally understand the structure-property relationships in luminescent AuNPs and apply them as contrast agents to address some critical challenges in bioimaging at both the in vitro and in vivo level. In Chapter 2, we described the synthesized ~20 nm polycrystalline AuNPs (pAuNPs), which successfully integrated and enhanced plasmonic and fluorescence properties into a single AuNP through the grain size effect. The combination of these properties in one NP enabled AuNPs to serve as a multimodal contrast agent for in vitro optical microscopic imaging, making it possible to develop correlative microscopic imaging techniques. In Chapters 3-5, we proposed a feasible approach to optimize the in vivo kinetics and clearance profile of nanoprobes for multimodality in vivo bioimaging applications by using straightforward surface chemistry with luminescent AuNPs as a model. Luminescent glutathione-coated AuNPs of ~2 nm were synthesized. Investigation of the biodistribution showed that these glutathione-coated AuNPs (GS-AuNPs) exhibit stealthiness to the reticuloendothelial system (RES) organs and efficient renal clearance, with only 3.7+/-1.9% and 0.3+/-0.1% accumulating in the liver and spleen, and over 65% of the injection dose cleared out via the urine within the first 72 hours. In addition, ~2.5 nm NIR-emitting radioactive glutathione-coated [198Au]AuNPs (GS-[198Au]AuNPs) were synthesized for further evaluation of the pharmacokinetic profile of GS-AuNPs and potential multimodal imaging. The results showed that the GS-[198Au]AuNPs behave like small-molecule contrast agents in

  19. Note: Radiochemical measurement of fuel and ablator areal densities in cryogenic implosions at the National Ignition Facility

    SciTech Connect

    Hagmann, C. Shaughnessy, D. A.; Moody, K. J.; Grant, P. M.; Gharibyan, N.; Gostic, J. M.; Wooddy, P. T.; Torretto, P. C.; Bandong, B. B.; Bionta, R.; Cerjan, C. J.; Bernstein, L. A.; Caggiano, J. A.; Sayre, D. B.; Schneider, D. H.; Henry, E. A.; Fortner, R. J.; Herrmann, H. W.; Knauer, J. P.

    2015-07-15

    A new radiochemical method for determining deuterium-tritium (DT) fuel and plastic ablator (CH) areal densities (ρR) in high-convergence, cryogenic inertial confinement fusion implosions at the National Ignition Facility is described. It is based on measuring the {sup 198}Au/{sup 196}Au activation ratio using the collected post-shot debris of the Au hohlraum. The Au ratio combined with the independently measured neutron down scatter ratio uniquely determines the areal densities ρR(DT) and ρR(CH) during burn in the context of a simple 1-dimensional capsule model. The results show larger than expected ρR(CH) values, hinting at the presence of cold fuel-ablator mix.

  20. Thermal neutron radiative capture cross-section of 186W(n, γ)187W reaction

    NASA Astrophysics Data System (ADS)

    Tan, V. H.; Son, P. N.

    2016-06-01

    The thermal neutron radiative capture cross section for 186W(n, γ)187W reaction was measured by the activation method using the filtered neutron beam at the Dalat research reactor. An optimal composition of Si and Bi, in single crystal form, has been used as neutron filters to create the high-purity filtered neutron beam with Cadmium ratio of Rcd = 420 and peak energy En = 0.025 eV. The induced activities in the irradiated samples were measured by a high resolution HPGe digital gamma-ray spectrometer. The present result of cross section has been determined relatively to the reference value of the standard reaction 197Au(n, γ)198Au. The necessary correction factors for gamma-ray true coincidence summing, and thermal neutron self-shielding effects were taken into account in this experiment by Monte Carlo simulations.

  1. Ingestion of iron in sow's faeces by piglets reared in farrowing crates with slotted floors.

    PubMed

    Gleed, P T; Sansom, B F

    1982-01-01

    1. Two sows were housed in farrowing crates with slotted floors and fed daily with approximately 200 microCi 198Au from 3 d before farrowing until their piglets were 21 d old. 2. Th sows' faeces became radioactive but their milk remained free of radioactivity. The piglets' mean (+/- SE) whole-body radioactivity was equivalent to 8.5 +/- 0.9 g (range 3.8-15.5 g) of faeces. This is probably a measure of their daily intake of faeces. 3. The possibility of using this natural coprophagia to prevent piglet anaemia has been demonstrated on two commercial piggeries by feeding the sows a diet containing 2000 mg Fe/kg.

  2. Non-destructive assay of mechanical components using gamma-rays and thermal neutrons

    SciTech Connect

    Souza, Erica Silvani; Avelino, Mila R.

    2013-05-06

    This work presents the results obtained in the inspection of several mechanical components through neutron and gamma-ray transmission radiography. The 4.46 Multiplication-Sign 10{sup 5} n.cm{sup -2}.s{sup -1} thermal neutron flux available at the main port of the Argonauta research reactor in Instituto de Engenharia Nuclear has been used as source for the neutron radiographic imaging. The 412 keV {gamma}-ray emitted by {sup 198}Au, also produced in that reactor, has been used as interrogation agent for the gamma radiography. Imaging Plates - IP specifically designed to operate with thermal neutrons or with X-rays have been employed as detectors and storage devices for each of these radiations.

  3. An Am/Be neutron source and its use in integral tests of differential neutron reaction cross-section data.

    PubMed

    Uddin, M S; Zaman, M R; Hossain, S M; Spahn, I; Sudár, S; Qaim, S M

    2010-09-01

    An Am/Be neutron source, installed recently at the Rajshahi University, is described. Neutron flux mapping was done using the nuclear reactions (197)Au(n,gamma)(198)Au, (113)In(n,gamma)(114m)In, (115)In(n,n'gamma)(115m)In and (58)Ni(n,p)(58)Co. An approximate validation of the neutron spectral shape was done using five neutron threshold detectors and the iterative unfolding code SULSA. Integral cross sections of the reactions (54)Fe(n,p)(54)Mn, (59)Co(n,p)(59)Fe and (92)Mo(n,p)(92m)Nb were measured with fast neutrons (E(n)>1.5MeV) and compared with data calculated using the neutron spectral distribution and the excitation function of each reaction given in data libraries: an agreement within +/-6% was found.

  4. Note: Radiochemical measurement of fuel and ablator areal densities in cryogenic implosions at the National Ignition Facility

    NASA Astrophysics Data System (ADS)

    Hagmann, C.; Shaughnessy, D. A.; Moody, K. J.; Grant, P. M.; Gharibyan, N.; Gostic, J. M.; Wooddy, P. T.; Torretto, P. C.; Bandong, B. B.; Bionta, R.; Cerjan, C. J.; Bernstein, L. A.; Caggiano, J. A.; Herrmann, H. W.; Knauer, J. P.; Sayre, D. B.; Schneider, D. H.; Henry, E. A.; Fortner, R. J.

    2015-07-01

    A new radiochemical method for determining deuterium-tritium (DT) fuel and plastic ablator (CH) areal densities (ρR) in high-convergence, cryogenic inertial confinement fusion implosions at the National Ignition Facility is described. It is based on measuring the 198Au/196Au activation ratio using the collected post-shot debris of the Au hohlraum. The Au ratio combined with the independently measured neutron down scatter ratio uniquely determines the areal densities ρR(DT) and ρR(CH) during burn in the context of a simple 1-dimensional capsule model. The results show larger than expected ρR(CH) values, hinting at the presence of cold fuel-ablator mix.

  5. {sup 7}Be in Stars and in the Laboratory

    SciTech Connect

    Hass, Michael; Kumar, Vivek

    2008-01-24

    We discuss results and future plans for low-energy reactions that play an important role in current nuclear astrophysics research and that happen to concentrate around the region of A = 7. The {sup 7}Be(p,{gamma}){sup 8}B and the {sup 3}He({sup 4}He,{gamma}){sup 7}Be reactions are crucial for understanding the solar-neutrino oscillations phenomenon and the latter one plays a central role in the issue of cosmic {sup 7}Li abundance and Big-Bang Nucleosynthesis. The electron-capture (EC) decay rate of {sup 7}Be in metallic Cu host and the {beta}{sup -}decay rate of {sup 198}Au in the host alloy Al-Au have been measured simultaneously at several temperatures, ranging from 0.350 K to 293 K. The resulting null temperature dependence is discussed in terms of the inadequacy of the often-used Debye-Hueckel model for such measurements.

  6. Age-specific inhalation radiation dose commitment factors for selected radionuclides

    SciTech Connect

    Strenge, D.L.; Peloquin, R.A.; Baker, D.A.

    1982-08-01

    Inhalation dose commitment factors are presented for selected radionuclides for exposure of individuals in four age groups: infant, child, teen and adult. Radionuclides considered are /sup 35/S, /sup 36/Cl, /sup 45/Ca, /sup 67/Ga, /sup 75/Se, /sup 85/Sr, /sup 109/Cd, /sup 113/Sn, /sup 125/I, /sup 133/Ba, /sup 170/Tm, /sup 169/Yb, /sup 182/Ta, /sup 192/Ir, /sup 198/Au, /sup 201/Tl, /sup 204/Tl, and /sup 236/Pu. The calculational method is based on the human metabolic model of ICRP as defined in Publication 2 (ICRP 1959) and as used in previous age-specific dose factor calculations by Hoenes and Soldat (1977). Dose commitment factors are presented for the following organs of reference: total body, bone, liver, kidney, thyroid, lung and lower large intestine.

  7. Pinocytosis in the rat visceral yolk sac. Effects of temperature, metabolic inhibitors and some other modifiers.

    PubMed

    Duncan, R; Lloyd, J B

    1978-12-18

    Low temperature,2,4-dinitrophenol and moniodoacetate could each completely abolish the pinocytic uptake of 125I-labelled polyvinylpyrrolidone, 125I-labelled bovine serum albumin or colloidal 198 Au by 17.5-day rat visceral yolk sac cultured in vitro. Cytochalasin B and colchicine caused a partial and dose-dependent inhibition. It is concluded that the mechanism of pinocytic uptake of these substrates is not micropinocytosis as conventionally defined. Removal of extracellular calcium or the presence of theophylline inhibited liquid-phase pinocytosis by the rat yolk sac, whereas addition of ouabain caused a biphasic response: a slight stimulation of pinosome formation at a low concentration, and an inhibitory effect at a higher concentration.

  8. K{sub Air} and H*(10) Rate Constants for Gamma Emitters

    SciTech Connect

    Vega-Carrillo, H. R.; Juarez, R. Rodriguez; Manzanares-Acuna, E.; Davila, V. M. Hernandez; Mercado, G. A.

    2008-08-11

    Monte Carlo calculations have been carried out to estimate the Air Kerma rate constant and the Ambient dose equivalent rate constant for 139 monoenergetic photon sources. The factor that relates activity to air kerma rate or to ambient dose equivalent is useful to estimate the dose from a photon emitter source. Here 139 point-like and monoenergetic gamma-ray sources, ranging from 0.01 to 10 MeV were utilized in Monte Carlo calculations to estimate both gamma factors. These factors were utilized to calculate the air kerma-and-ambient dose equivalent rate constants for {sup 137}Cs-{sup 137m}Ba, {sup 198}Au, {sup 60}Co, and {sup 131}I, whose values were compared with those published in the literature.

  9. 232Th(n,γ)233Th Thermal Reaction Cross-Section Measurement

    NASA Astrophysics Data System (ADS)

    Maidana, Nora L.; Vanin, Vito R.; Castro, Ruy M.; Pascholati, Paulo R.; Helene, Otaviano; Dias, Mauro S.; Koskinas, Marina F.

    2005-05-01

    The 232Th(n,γ)233Th thermal neutron-capture reaction cross section was measured using targets of ˜ 1.5 mg of high-purity metallic thorium irradiated in the IPEN IEA-R1m 5 MW pool research reactor. The 197Au(n,γ)198Au reaction was used to monitor the thermal and epithermal neutron fluxes in the irradiation position, which was found using the Westcott formalism. The residual gamma-ray activity was followed with an HPGe detector. The detector efficiency curve was fitted by the least-squares method applying covariance analysis to all uncertainties involved. The experimental result is σ0 =7.20±0.20 b, in agreement with previous published values.

  10. Note: Radiochemical measurement of fuel and ablator areal densities in cryogenic implosions at the National Ignition Facility.

    PubMed

    Hagmann, C; Shaughnessy, D A; Moody, K J; Grant, P M; Gharibyan, N; Gostic, J M; Wooddy, P T; Torretto, P C; Bandong, B B; Bionta, R; Cerjan, C J; Bernstein, L A; Caggiano, J A; Herrmann, H W; Knauer, J P; Sayre, D B; Schneider, D H; Henry, E A; Fortner, R J

    2015-07-01

    A new radiochemical method for determining deuterium-tritium (DT) fuel and plastic ablator (CH) areal densities (ρR) in high-convergence, cryogenic inertial confinement fusion implosions at the National Ignition Facility is described. It is based on measuring the (198)Au/(196)Au activation ratio using the collected post-shot debris of the Au hohlraum. The Au ratio combined with the independently measured neutron down scatter ratio uniquely determines the areal densities ρR(DT) and ρR(CH) during burn in the context of a simple 1-dimensional capsule model. The results show larger than expected ρR(CH) values, hinting at the presence of cold fuel-ablator mix.

  11. [Studies on the lymph drainage of the eye. 2. Drainage of lymph-dependent radioactive substances from the anterior chamber. (author's transl)].

    PubMed

    Grüntzig, J; Schicha, H; Becker, V; Kiem, J; Feinendegen, L E

    1977-10-01

    In order to study lymph drainage from the eye colloid solutions (198Au-colloid and 99mTc-sulfur-colloid) were injected into the right anterior chamber of the eye in 12 rabbits. The distribution of the radioactivity was measured in vivo with an Anger-type camera and in vitro with a sodium iodine crystal well-counter after dissection. The drainage of radioactivity from the anterior chamber had an average half-life of 36 min. The drained activity could be found mainly in blood, liver, kidney, and bone tissue. Moreover, a concentration of radioactivity significantly surpassing the concentration in blood was measured in the right retrobulbar space in 75% of the cases and in the cervical lymph nodes in 50% of the cases. These findings substantiate a communication between the anterior chamber of the eye and the retrobulbar space in the form of a lymphatic or a prelymphatic-lymphatic flow.

  12. Blood clearance of radiolabeled gold colloid by the turkey mononuclear phagocytic system.

    PubMed

    McEntee, M F; Ficken, M D

    1990-01-01

    Radiolabeled gold colloid (198Au), which has been used to assess particle clearance in mammalian species, was used to assess blood-borne particle clearance in turkeys. When turkeys 16 weeks of age were injected intravenously with these particles, there was greater than a 98% decrease in blood gamma emission from 1 minute to 6 minutes postinjection. Uptake of particles was predominantly hepatic with minor uptake by the spleen and bone marrow. Negligible uptake was observed in lung, kidney, and skeletal muscle. Autoradiography demonstrated particles within Kupffer cells of the liver, periarteriolar macrophages of the spleen, and bone-marrow macrophages. Particles could not be demonstrated within the lung or kidney. The mononuclear phagocyte system responsible for blood particle clearance in turkeys is therefore located predominantly within the liver, spleen, and bone marrow and is similar to that of rats, mice, rabbits, and dogs. Pulmonary intravascular macrophages, which have recently been described in ruminants and pigs, are not apparent in this species.

  13. Measurements and analyses of the distribution of the radioactivity induced by the secondary neutrons produced by 17-MeV protons in compact cyclotron facility

    NASA Astrophysics Data System (ADS)

    Matsuda, Norihiro; Izumi, Yuichi; Yamanaka, Yoshiyuki; Gandou, Toshiyuki; Yamada, Masaaki; Oishi, Koji

    2017-09-01

    Measurements of reaction rates by secondary neutrons produced from beam losses by 17-MeV protons are conducted at a compact cyclotron facility with the foil activation method. The experimentally obtained distribution of the reaction rates of 197Au (n, γ) 198Au on the concrete walls suggests that a target and an electrostatic deflector as machine components for beam extraction of the compact cyclotron are principal beam loss points. The measurements are compared with calculations by the Monte Carlo code: PHITS. The calculated results based on the beam losses are good agreements with the measured ones within 21%. In this compact cyclotron facility, exponential attenuations with the distance from the electrostatic deflector in the distributions of the measured reaction rates were observed, which was looser than that by the inverse square of distance.

  14. Gold(III) complexes in medicinal chemistry.

    PubMed

    Maia, Pedro Ivo da Silva; Deflon, Victor M; Abram, Ulrich

    2014-09-01

    A number of gold(III) compounds has been designed with the objective of overcoming the disadvantages associated with the platinum-based drugs for cancer treatment. Compounds of a remarkable structural manifold show significant antiproliferative effects in vitro against a number of cancer cells, including cisplatin resistant ones. The target of most of them is, unlike that of cisplatin, not the DNA. Although the mechanisms of action displayed by the gold compounds in biological media are still under investigation, many studies show evidence that the cellular targets are mitochondria-based. Recent advances in gold(III) medicinal chemistry also recommend such compounds for other pharmacological applications such as the treatment of viral or parasitic diseases. The radioactive isotopes (198)Au and (199)Au present potential in radiotherapy.

  15. Optimised k0-instrumental neutron activation method using the TRIGA MARK I IPR-R1 reactor at CDTN/CNEN, Belo Horizonte, Brazil

    NASA Astrophysics Data System (ADS)

    Menezes, M. Â. B. C.; Jaćimović, R.

    2006-08-01

    The Nuclear Technology Development Centre/Brazilian Commission for Nuclear Energy, CDTN/CNEN, is the only Brazilian Institution to apply the k0-standardisation method of instrumental neutron activation technique determining elements using its own nuclear reactor, TRIGA MARK I IPR-R1. After changes in the reactor core configuration, the reactor neutron flux distribution in typical irradiation channels had to be updated, as well as the parameters f and α, needed to apply the k0-method of neutron activation analysis. The neutron flux distribution in the rotary rack was evaluated through the specific count rate of 198Au and the parameters f and α, were determined in five selected channels applying the "Cd-ratio for multi-monitor" method, using a set of Al-(0.1%)Au and Zr (99.8%) monitors. Several reference materials were analysed, indicating the effectiveness of the improved method.

  16. 232Th(n,{gamma})233Th Thermal Reaction Cross-Section Measurement

    SciTech Connect

    Maidana, Nora L.; Vanin, Vito R.; Pascholati, Paulo R.; Helene, Otaviano; Castro, Ruy M.; Dias, Mauro S.; Koskinas, Marina F.

    2005-05-24

    The 232Th(n,{gamma})233Th thermal neutron-capture reaction cross section was measured using targets of {approx} 1.5 mg of high-purity metallic thorium irradiated in the IPEN IEA-R1m 5 MW pool research reactor. The 197Au(n,{gamma})198Au reaction was used to monitor the thermal and epithermal neutron fluxes in the irradiation position, which was found using the Westcott formalism. The residual gamma-ray activity was followed with an HPGe detector. The detector efficiency curve was fitted by the least-squares method applying covariance analysis to all uncertainties involved. The experimental result is {sigma}0 =7.20{+-}0.20 b, in agreement with previous published values.

  17. Level densities and thermodynamical properties of Pt and Au isotopes

    NASA Astrophysics Data System (ADS)

    Giacoppo, F.; Bello Garrote, F. L.; Bernstein, L. A.; Bleuel, D. L.; Eriksen, T. K.; Firestone, R. B.; Görgen, A.; Guttormsen, M.; Hagen, T. W.; Kheswa, B. V.; Klintefjord, M.; Koehler, P. E.; Larsen, A. C.; Nyhus, H. T.; Renstrøm, T.; Sahin, E.; Siem, S.; Tornyi, T.

    2014-11-01

    The nuclear level densities of Pt-196194 and Au,198197 below the neutron separation energy have been measured using transfer and scattering reactions. All the level density distributions follow the constant-temperature description. Each group of isotopes is characterized by the same temperature above the energy threshold corresponding to the breaking of the first Cooper pair. A constant entropy excess Δ S =1.9 kB and 1.1 kB is observed in 195Pt and 198Au with respect to 196Pt and 197Au, respectively, giving information on the available single-particle level space for the last unpaired valence neutron. The breaking of nucleon Cooper pairs is revealed by sequential peaks in the microcanonical caloric curve.

  18. Whole-body imaging of the distribution of mercury released from dental fillings into monkey tissues

    SciTech Connect

    Hahn, L.J.; Kloiber, R.; Leininger, R.W.; Vimy, M.J.; Lorscheider, F.L. )

    1990-11-01

    The fate of mercury (Hg) released from dental silver amalgam tooth fillings into human mouth air is uncertain. A previous report about sheep revealed uptake routes and distribution of amalgam Hg among body tissues. The present investigation demonstrates the bodily distribution of amalgam Hg in a monkey whose dentition, diet, feeding regimen, and chewing pattern closely resemble those of humans. When amalgam fillings, which normally contain 50% Hg, are made with a tracer of radioactive {sup 203}Hg and then placed into monkey teeth, the isotope appears in high concentration in various organs and tissues within 4 wk. Whole-body images of the monkey revealed that the highest levels of Hg were located in the kidney, gastrointestinal tract, and jaw. The dental profession's advocacy of silver amalgam as a stable tooth restorative material is not supported by these findings.

  19. Uptake of Mercury by the Brain

    PubMed Central

    Magos, L.

    1968-01-01

    A technique has been developed for injecting metallic mercury intravenously in aqueous solution. Thirty seconds after intravenous injection of rats with 0·1 μg. metallic mercury labelled with 203Hg nearly 20% of the dose had been exhaled and the concentration in the brain was nearly as high as in the blood. After injection of mercuric ion little of the dose was exhaled, and brain uptake was much less. Oxidation of mercury in the blood was, therefore, not instantaneous, and the rapid transport of the unconverted metallic mercury to the brain and its subsequent rapid diffusion from the blood was responsible for the high level of mercury in the brain after exposure to mercury vapour. The technique might be useful for the study of the passage of highly diffusible vapours through the respiratory membranes. PMID:5723353

  20. Methods for measuring specific rates of mercury methylation and degradation and their use in determining factors controlling net rates of mercury methylation

    SciTech Connect

    Ramlal, P.S.; Rudd, J.W.M.; Hecky, R.E.

    1986-01-01

    A method was developed to estimate specific rates of demethylation of methyl mercury in aquatic samples by measuring the volatile /sup 14/C end products of /sup 14/CH/sub 3/HgI demethylation. This method was used in conjuction with a /sup 203/Hg/sup 2 +/ radiochemical method which determines specific rates of mercury methylation. Together, these methods enabled us to examine some factors controlling the net rate of mercury methylation. The methodologies were field tested, using lake sediment samples from a recently flooded reservoir in the Southern Indian Lake system which had developed a mercury contamination problem in fish. Ratios of the specific rates of methylation/demethylation were calculated. The highest ratios of methylation/demethylation were calculated. The highest ratios of methylation/demethylation occurred in the flooded shorelines of Southern Indian Lake. These results provide an explanation for the observed increases in the methyl mercury concentrations in fish after flooding.

  1. Experimental cross-sections for proton induced nuclear reactions on mercury up to 65 MeV

    NASA Astrophysics Data System (ADS)

    Hermanne, A.; Tárkányi, F.; Takács, S.; Ditrói, F.; Szücs, Z.; Brezovcsik, K.

    2016-07-01

    Cross-sections for formation of activation products induced by protons on natural mercury targets were measured. Results for 196m,196g,197g(cum), 198m,198g,199g(cum), 200g(cum), 201,202Tl, 194g(cum), 195g(cum), 196g(cum), 198m,199g(cum) Au and 195m,197m,203Hg are presented up to 65 MeV incident particle energy, many of these for the first time. The experimental data are compared with literature values and with the predictions of the TALYS 1.6 code (results taken from TENDL-2015 on-line library), thick target yields were derived and possible applications in biomedical sciences are discussed.

  2. Mercuric reductase activity and evidence of broad-spectrum mercury resistance among clinical isolates of rapidly growing mycobacteria

    SciTech Connect

    Steingrube, V.A.; Wallace, R.J. Jr.; Steele, L.C.; Pang, Y.J. )

    1991-05-01

    Resistance to mercury was evaluated in 356 rapidly growing mycobacteria belonging to eight taxonomic groups. Resistance to inorganic Hg2+ ranged from 0% among the unnamed third biovariant complex of Mycobacterium fortuitum to 83% among M. chelonae-like organisms. With cell extracts and 203Hg(NO3)2 as the substrate, mercuric reductase (HgRe) activity was demonstrable in six of eight taxonomic groups. HgRe activity was inducible and required NADPH or NADH and a thiol donor for optimai activity. Species with HgRe activity were also resistant to organomercurial compounds, including phenylmercuric acetate. Attempts at intraspecies and intragenus transfer of HgRe activity by conjugation or transformation were unsuccessful. Mercury resistance is common in rapidly growing mycobacteria and appears to function via the same inducible enzyme systems already defined in other bacterial species. This system offers potential as a strain marker for epidemiologic investigations and for studying genetic systems in rapidly growing mycobacteria.

  3. Thermal neutron capture and resonance integral cross sections of 45Sc

    NASA Astrophysics Data System (ADS)

    Van Do, Nguyen; Duc Khue, Pham; Tien Thanh, Kim; Thi Hien, Nguyen; Kim, Guinyun; Kim, Kwangsoo; Shin, Sung-Gyun; Cho, Moo-Hyun; Lee, Manwoo

    2015-11-01

    The thermal neutron cross section (σ0) and resonance integral (I0) of the 45Sc(n,γ)46Sc reaction have been measured relative to that of the 197Au(n,γ)198Au reaction by means of the activation method. High-purity natural scandium and gold foils without and with a cadmium cover of 0.5 mm thickness were irradiated with moderated pulsed neutrons produced from the Pohang Neutron Facility (PNF). The induced activities in the activated foils were measured with a high purity germanium (HPGe) detector. In order to improve the accuracy of the experimental results the counting losses caused by the thermal (Gth) and resonance (Gepi) neutron self-shielding, the γ-ray attenuation (Fg) and the true γ-ray coincidence summing effects were made. In addition, the effect of non-ideal epithermal spectrum was also taken into account by determining the neutron spectrum shape factor (α). The thermal neutron cross-section and resonance integral of the 45Sc(n,γ)46Sc reaction have been determined relative to the reference values of the 197Au(n,γ)198Au reaction, with σo,Au = 98.65 ± 0.09 barn and Io,Au = 1550 ± 28 barn. The present thermal neutron cross section has been determined to be σo,Sc = 27.5 ± 0.8 barn. According to the definition of cadmium cut-off energy at 0.55 eV, the present resonance integral cross section has been determined to be Io,Sc = 12.4 ± 0.7 barn. The present results are compared with literature values and discussed.

  4. Manufacturing techniques studies of ceramics by neutron and γ-ray radiography

    NASA Astrophysics Data System (ADS)

    Latini, R. M.; Souza, M. I. S.; Almeida, G. L.; Bellido, A. V. B.

    2014-11-01

    In this study, the aim was to evaluate capabilities and constraints of radiographic imagery using thermal neutrons and gamma-rays as tools to identify the type of technique employed in ceramics manufacturing especially that used in prehistoric Brazilian pottery from Acre state. For this purpose, radiographic images of test objects made with clay of this region using both techniques - palette and rollers - have been acquired with a system comprised of a source of gamma-rays or thermal neutrons and a corresponding X-ray or neutron-sensitive Imaging Plate as detector. For the neutrongraphy samples were exposed to a thermal neutron flux of order of 105n.cm-2.s-1 for 3 minutes at main port of Argonauta research reactor of the Instituto de Engenharia Nuclear - IEN/CNEN. The radiographic images using γ-rays from 165Dy (95 keV) and 198Au (412 keV) both produced at this reactor, have been acquired under an exposure time of a couple of hours. After acquisition, images have undergone a treatment to improve their quality through enhancement of their contrast, a procedure involving corrections of the beam divergence, sample shape and averaging of the attenuation map profile. Preliminary results show that difference between manufacturing techniques is better identified by radiography using low energy γ-rays from 165Dy rather than neutrongraphy or γ-rays from 198Au . Nevertheless, disregarding the kind of employed radiation, it should be stressed that feasibility to apply the technique is tightly tied to homogeneity of the clay itself and tempers due to their different attenuation.

  5. Manufacturing techniques studies of ceramics by neutron and γ-ray radiography

    SciTech Connect

    Latini, R. M.; Bellido, A. V. B.; Souza, M. I. S.; Almeida, G. L.

    2014-11-11

    In this study, the aim was to evaluate capabilities and constraints of radiographic imagery using thermal neutrons and gamma-rays as tools to identify the type of technique employed in ceramics manufacturing especially that used in prehistoric Brazilian pottery from Acre state. For this purpose, radiographic images of test objects made with clay of this region using both techniques - palette and rollers - have been acquired with a system comprised of a source of gamma-rays or thermal neutrons and a corresponding X-ray or neutron-sensitive Imaging Plate as detector. For the neutrongraphy samples were exposed to a thermal neutron flux of order of 10{sup 5}n.cm{sup −2}.s{sup −1} for 3 minutes at main port of Argonauta research reactor of the Instituto de Engenharia Nuclear - IEN/CNEN. The radiographic images using γ-rays from {sup 165}Dy (95 keV) and {sup 198}Au (412 keV) both produced at this reactor, have been acquired under an exposure time of a couple of hours. After acquisition, images have undergone a treatment to improve their quality through enhancement of their contrast, a procedure involving corrections of the beam divergence, sample shape and averaging of the attenuation map profile. Preliminary results show that difference between manufacturing techniques is better identified by radiography using low energy γ-rays from {sup 165}Dy rather than neutrongraphy or γ-rays from {sup 198}Au. Nevertheless, disregarding the kind of employed radiation, it should be stressed that feasibility to apply the technique is tightly tied to homogeneity of the clay itself and tempers due to their different attenuation.

  6. Mercury methylation and sulfate reduction rates in mangrove sediments, Rio de Janeiro, Brazil: The role of different microorganism consortia.

    PubMed

    Correia, Raquel Rose Silva; Guimarães, Jean Remy Davée

    2017-01-01

    Recent studies have shown Hg methylation in mangrove sediments, however, little is known about the different microorganism consortia involved. We investigated the participation of prokaryotes in general, iron-reducing bacteria-IRB, sulfate-reducing bacteria-SRB, methanogens and fungi in Hg methylation and sulfate reduction rates (SRR) in mangrove sediments using iron amendments for IRB and specific inhibitors for the other microorganisms. Sediment samples were collected from two mangrove zones, tidal flat and mangrove forest (named root sediments). Samples were incubated with (203)Hg or (35)SO4(2-) and Me(203)Hg/(35)Sulfur were measured by liquid scintillation. Methylmercury (MeHg) formation was significantly reduced when SRB (87.7%), prokaryotes (76%) and methanogens (36.5%) were inhibited in root sediments, but only SRB (51.6%) and prokaryotes (57.3%) in tidal flat. However, in the tidal flat, inhibition of methanogens doubled Hg methylation (104.5%). All inhibitors (except fungicide) significantly reduced SRR in both zones. In iron amended tidal flat samples, Hg methylation increased 56.5% at 100 μg g(-1) and decreased at 500 and 1000 μg g(-1) (57.8 and 82%). In the roots region, however, MeHg formation gradually decreased in response to Fe amendments from 100 μg g(-1) (37.7%) to 1000 μg g(-1) (93%). SRR decreased in all iron amendments. This first simultaneous evaluation of Hg methylation and sulfate-reduction and of the effect of iron and inhibitors on both processes suggest that SRB are important Hg methylators in mangrove sediments. However, it also suggests that SRB activity could not explain all MeHg formation. This implies the direct or indirect participation of other microorganisms such as IRB and methanogens and a complex relationship among these groups.

  7. MRP2 and the Handling of Mercuric Ions in Rats Exposed Acutely to Inorganic and Organic Species of Mercury

    PubMed Central

    Bridges, Christy C.; Joshee, Lucy; Zalups, Rudolfs K.

    2011-01-01

    Mercuric ions accumulate preferentially in renal tubular epithelial cells and bond with intracellular thiols. Certain metal-complexing agents have been shown to promote extraction of mercuric ions via the multidrug resistance-associated protein 2 (MRP2). Following exposure to a non-toxic dose of inorganic mercury (Hg2+), in the absence of complexing agents, tubular cells are capable of exporting a small fraction of intracellular Hg2+ through one or more undetermined mechanisms. We hypothesize that MRP2 plays a role in this export. To test this hypothesis, Wistar (control) and TR− rats were injected intravenously with a non-nephrotoxic dose of HgCl2 (0.5 μmol/kg) or CH3HgCl (5 mg/kg), containing [203Hg], in the presence or absence of cysteine (Cys; 1.25 μmol/kg or 12.5 mg/kg, respectively). Animals were sacrificed 24 h after exposure to mercury and the content of [203Hg] in blood, kidneys, liver, urine and feces was determined. In addition, uptake of Cys-S-conjugates of Hg2+ and methylmercury (CH3Hg+) was measured in inside-out membrane vesicles prepared from either control Sf9 cells or Sf9 cells transfected with human MRP2. The amount of mercury in the total renal mass and liver was significantly greater in TR− rats than in controls. In contrast, the amount of mercury in urine and feces was significantly lower in TR− rats than in controls. Data from membrane vesicles indicate that Cys-S-conjugates of Hg2+ and CH3Hg+ are transportable substrates of MRP2. Collectively, these data indicate that MRP2 plays a role in the physiological handling and elimination of mercuric ions from the kidney. PMID:21134393

  8. Slow biliary elimination of methyl mercury in the marine elasmobranchs, Raja erinacea and Squalus acanthias.

    PubMed

    Ballatori, N; Boyer, J L

    1986-09-30

    The present study examined the ability of two marine elasmobranchs (Raja erinacea, little skate, and Squalus acanthias, spiny dogfish shark) to excrete methyl mercury into bile, a major excretory route in mammals. 203Hg-labeled methyl mercury chloride was administered via the caudal vein, and bile collected through exteriorized cannulas in the free swimming fish. Skates and dogfish sharks excreted only a small fraction of the 203Hg into bile over a 3-day period: in the skate, the 3-day cumulative excretion (as a % of dose) was 0.44 +/- 0.10 (n = 4, +/- SD), 0.71 +/- 0.23 (n = 6), and 1.00 +/- 0.34(n = 4) for doses of 1, 5, and 20 mumol/kg, respectively, while the shark excreted only 0.15 +/- 0.15% (n = 8) at a dose of 5 mumol/kg. As in mammals, the availability of hepatic and biliary glutathione was a determinant of the biliary excretion of methyl mercury in these species: the administration of sulfobromophthalein, a compound known to inhibit both glutathione and methyl mercury excretion in rats, or of L-buthionine-S,R-sulfoximine, an inhibitor of glutathione biosynthesis, decreased the biliary excretion of both glutathione and mercury in the skate. The slow hepatic excretory process for methyl mercury in the skate and shark was attributed to an inordinately slow rate of bile formation: from 1 to 4 ml/kg X day. An inefficient biliary excretory process in fish may account in part for the long biological half-times for methyl mercury in marine species.

  9. Comparative effects of chelating agents on distribution, excretion, and renal toxicity of inorganic mercury in rats

    SciTech Connect

    Kojima, S.; Shimada, H.; Kiyozumi, M. )

    1989-06-01

    The effects of three chelating agents, sodium N-benzyl-D-glucamine dithiocarbamate(NBG-DTC), 2,3-dimercaptopropanol(BAL), and D-penicillamine(D-PEN), on the distribution, excretion, and renal toxicity of inorganic mercury were compared in rats exposed to HgCl2. Rats were injected i.p. with 203HgCl2 (300 micrograms of Hg and 2 microCi of 203Hg/kg) and 30 min or 24 h later they were injected with a chelating agent (a quarter of an LD50). The injection of the chelating agents significantly enhanced the biliary and urinary excretions of mercury. BAL was the most effective for removal of mercury from the body at 30 min after mercury treatment. The extent of enhancing effect of the chelating agents for removal of mercury at 24 h after mercury was in the order NBG-DTC = BAL greater than D-PEN. The injection of BAL at 24 h after mercury treatment caused the redistribution of mercury to the heart and lung. NBG-DTC did not result in the redistribution of mercury to the heart, lung, and brain. Urinary excretion of protein and AST significantly increased 24-48 h after mercury treatment and decreased to the control values 72 h after mercury. The injection of the chelating agents at 30 min after mercury treatment significantly decreased the urinary excretion of protein and AST. In rats pretreated with mercury 24 h earlier, the chelating agents significantly decreased the urinary protein at 48 h after mercury treatment, but did not decrease the urinary AST. The results of this study indicate that the chelating agents are effective in removing mercury from the body, resulting in the protective effect against the mercury-induced renal damage.

  10. MRP2 and the handling of mercuric ions in rats exposed acutely to inorganic and organic species of mercury

    SciTech Connect

    Bridges, Christy C. Joshee, Lucy; Zalups, Rudolfs K.

    2011-02-15

    Mercuric ions accumulate preferentially in renal tubular epithelial cells and bond with intracellular thiols. Certain metal-complexing agents have been shown to promote extraction of mercuric ions via the multidrug resistance-associated protein 2 (MRP2). Following exposure to a non-toxic dose of inorganic mercury (Hg{sup 2+}), in the absence of complexing agents, tubular cells are capable of exporting a small fraction of intracellular Hg{sup 2+} through one or more undetermined mechanisms. We hypothesize that MRP2 plays a role in this export. To test this hypothesis, Wistar (control) and TR{sup -} rats were injected intravenously with a non-nephrotoxic dose of HgCl{sub 2} (0.5 {mu}mol/kg) or CH{sub 3}HgCl (5 mg/kg), containing [{sup 203}Hg], in the presence or absence of cysteine (Cys; 1.25 {mu}mol/kg or 12.5 mg/kg, respectively). Animals were sacrificed 24 h after exposure to mercury and the content of [{sup 203}Hg] in blood, kidneys, liver, urine and feces was determined. In addition, uptake of Cys-S-conjugates of Hg{sup 2+} and methylmercury (CH{sub 3}Hg{sup +}) was measured in inside-out membrane vesicles prepared from either control Sf9 cells or Sf9 cells transfected with human MRP2. The amount of mercury in the total renal mass and liver was significantly greater in TR{sup -} rats than in controls. In contrast, the amount of mercury in urine and feces was significantly lower in TR{sup -} rats than in controls. Data from membrane vesicles indicate that Cys-S-conjugates of Hg{sup 2+} and CH{sub 3}Hg{sup +} are transportable substrates of MRP2. Collectively, these data indicate that MRP2 plays a role in the physiological handling and elimination of mercuric ions from the kidney.

  11. Phospholipase D catalyzes phospholipid metabolism in chemotactic peptide-stimulated HL-60 granulocytes

    SciTech Connect

    Pai, J.K.; Siegel, M.I.; Egan, R.W.; Billah, M.M.

    1988-09-05

    There exists circumstantial evidence for activation of phospholipase D (PLD) in intact cells. However, because of the complexity of phospholipid remodeling processes, it is essential to distinguish PLD clearly from other phospholipases and phospholipid remodeling enzymes. Therefore, to establish unequivocally PLD activity in dimethyl sulfoxide-differentiated HL-60 granulocytes, to demonstrate the relative contribution of PLD to phospholipid turnover, and to validate the hypothesis that the formation of phosphatidylethanol is an expression of PLD-catalyzed transphosphatidylation, we have developed methodologies to label HL-60 granulocytes in 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholine (alkyl-PC) with 32P without labeling cellular ATP. These methodologies involve (a) synthesis of alkyl-lysoPC containing 32P by a combination of enzymatic and chemical procedures and (b) incubation of HL-60 granulocytes with this alkyl-(32P) lysoPC which enters the cell and becomes acylated into membrane-associated alkyl-(32P)PC. Upon stimulation of these 32P-labeled cells with the chemotactic peptide, N-formyl-Met-Leu-Phe (fMLP), alkyl-(32P)phosphatidic acid (alkyl-(32P)PA) is formed rapidly. Because, under these conditions, cellular ATP has not been labeled with 32P, alkyl-(32P)PA must be formed via PLD-catalyzed hydrolysis of alkyl-(32P)PC at the terminal phosphodiester bond. This result conclusively demonstrates fMLP-induced activation of PLD in HL-60 granulocytes. These 32P-labeled HL-60 granulocytes have also been stimulated in the presence of ethanol to produce alkyl-(32P)phosphatidylethanol (alkyl-(32P)PEt). Formation of alkyl-(32P)PEt parallels that of alkyl-(32P)PA with respect to time course, fMLP concentration, inhibition by a specific fMLP antagonist (t-butoxycarbonyl-Met-Leu-Phe), and Ca2+ concentration.

  12. Measurement of the 115In(n,γ)116 m In reaction cross-section at the neutron energies of 1.12, 2.12, 3.12 and 4.12 MeV

    NASA Astrophysics Data System (ADS)

    Lawriniang, Bioletty Mary; Badwar, Sylvia; Ghosh, Reetuparna; Jyrwa, Betylda; Vansola, Vibha; Naik, Haladhara; Goswami, Ashok; Naik, Yeshwant; Datrik, Chandra Shekhar; Gupta, Amit Kumar; Singh, Vijay Pal; Pol, Sudir Shibaji; Subramanyam, Nagaraju Balabenkata; Agarwal, Arun; Singh, Pitambar

    2015-08-01

    The 115In(n,γ)116 m In reaction cross section at neutron energies of 1.12, 2.12, 3.12 and 4.12 MeV was determined by using an activation and off-line γ-ray spectrometric technique. The monoenergetic neutron energies of 1.12 - 4.12 MeV were generated from the 7Li(p,n) reaction by using proton beam with energies of 3 and 4 MeV from the folded tandem ion beam accelerator (FOTIA) at Bhabha Atomic Research Centre (BARC) and with energies of 5 and 6 MeV from the Pelletron facility at Tata Institute of Fundamental Research (TIFR), Mumbai. The 197Au(n,γ)198Au reaction cross-section was used as the neutron flux monitor.The 115In(n,γ)116 m In reaction cross section at neutron energies of 1.12, 2.12, 3.12 and 4.12 MeV was determined by using an activation and off-line γ-ray spectrometric technique. The monoenergetic neutron energies of 1.12 - 4.12 MeV were generated from the 7Li(p,n) reaction by using proton beam with energies of 3 and 4 MeV from the folded tandem ion beam accelerator (FOTIA) at Bhabha Atomic Research Centre (BARC) and with energies of 5 and 6 MeV from the Pelletron facility at Tata Institute of Fundamental Research (TIFR), Mumbai. The 197Au(n,γ)198 Au reaction cross-section was used as the neutron flux monitor. The 115In(n,γ)116 m In reaction cross-sections at neutron energies of 1.12 - 4.12 MeV were compared with the literature data and were found to be in good agreement with one set of data, but not with others. The 115In(n,γ)116 m In cross-section was also calculated theoretically by using the computer code TALYS 1.6 and was found to be slightly lower than the experimental data from the present work and the literature.)198Au reaction cross-section was used as the neutron flux monitor. The 115In(n,γ)116 m In reaction cross-sections at neutron energies of 1.12 - 4.12 MeV were compared with the literature data and were found to be in good agreement with one set of data, but not with others. The 115In(n,γ)116 m In cross-section was also calculated

  13. Long-range effect of cyanide on mercury methylation in a gold mining area in southern Ecuador.

    PubMed

    Guimaraes, Jean Remy Davée; Betancourt, Oscar; Miranda, Marcio Rodrigues; Barriga, Ramiro; Cueva, Edwin; Betancourt, Sebastián

    2011-11-01

    Small-scale gold mining in Portovelo-Zaruma, Southern Equador, performed by mercury amalgamation and cyanidation, yields 9-10 t of gold/annum, resulting in annual releases of around 0.65 t of inorganic mercury and 6000 t of sodium cyanide in the local river system. The release of sediments, cyanide, mercury, and other metals present in the ore such as lead, manganese and arsenic significantly reduces biodiversity downstream the processing plants and enriches metals in bottom sediments and biota. However, methylmercury concentrations in sediments downstream the mining area were recently found to be one order of magnitude lower than upstream or in small tributaries. In this study we investigated cyanide, bacterial activity in water and sediment and mercury methylation potentials in sediments along the Puyango river watershed, measured respectively by in-situ spectrophotometry and incubation with (3)H-leucine and (203)Hg(2+). Free cyanide was undetectable (<1 μg·L(-1)) upstream mining activities, reached 280 μg·L(-1) a few km downstream the processing plants area and was still detectable about 100 km downstream. At stations with detectable free cyanide in unfiltered water, 50% of it was dissolved and 50% associated to suspended particles. Bacterial activity and mercury methylation in sediment showed a similar spatial pattern, inverse to the one found for free cyanide in water, i.e. with significant values in pristine upstream sampling points (respectively 6.4 to 22 μgC·mg wet weight(-1)·h(-1) and 1.2 to 19% of total (203) Hg·gdry weight(-1)·day(-1)) and undetectable downstream the processing plants, returning to upstream values only in the most distant downstream stations. The data suggest that free cyanide oxidation was slower than would be expected from the high water turbulence, resulting in a long-range inhibition of bacterial activity and hence mercury methylation. The important mercury fluxes resultant from mining activities raise concerns about its

  14. Mercury net methylation in five tropical flood plain regions of Brazil: high in the root zone of floating macrophyte mats but low in surface sediments and flooded soils.

    PubMed

    Guimarães, J R; Meili, M; Hylander, L D; de Castro e Silva, E; Roulet, M; Mauro, J B; de Lemos, R

    2000-10-16

    In aquatic systems, bottom sediments have often been considered as the main methylmercury (MeHg) production site. In tropical floodplain areas, however, floating meadows and flooded forests extend over large areas and can be important Hg methylating sites. We present here a cross-system comparison of the Hg net methylation capacity in surface sediments, flooded soils and roots of floating aquatic macrophytes, assayed by in situ incubation with 203Hg and extraction of formed Me203 Hg by acid leaching and toluene. The presence of mono-MeHg was confirmed by thin layer chromatography and other techniques. Study areas included floodplain lakes in the Amazon basin (Tapajós, Negro and Amazon rivers), the Pantanal floodplain (Paraguay river basin), freshwater coastal lagoons in Rio de Janeiro and oxbow lakes in the Mogi-Guaçú river, São Paulo state. Different Hg levels were added in assays performed in 1994-1998, but great care was taken to standardise all other test parameters, to allow data comparisons. Net MeHg production was one order of magnitude higher (mean 13.8%, range 0.28-35) in the living or decomposing roots of floating or rooted macrophyte mats (Eichhornia azurea, E. crassipes, Paspalum sp., Eleocharis sellowiana, Salvinia sp., S. rotundifolia and Scirpus cubensis) than in the surface layer of underlying lake sediments (mean 0.6%, range 0.022-2.5). Methylation in flooded soils presented a wide range and was in some cases similar to the one found in macrophyte roots but usually much lower. In a Tapajós floodplain lake, natural concentrations of MeHg in soil and sediment cores taken along a lake-forest transect agreed well with data on net methylation potentials in the same samples. E. azurea, E. crassipes and Salvinia presented the highest methylation potentials, up to 113 times higher than in sediments. Methylation in E. azurea from six lakes of the Paraguay and Cuiabá rivers, high Pantanal, was determined in the 1998 dry and wet seasons and ranged from

  15. Endogenous ADP-ribosylation of elongation factor 2 in polyoma virus-transformed baby hamster kidney cells

    SciTech Connect

    Fendrick, J.L.; Iglewski, W.J. )

    1989-01-01

    Polyoma virus-transformed baby hamster kidney (pyBHK) cells were cultured in medium containing ({sup 32}P)orthophosphate and 105 (vol/vol) fetal bovine serum. A {sup 32}P-labeled protein with an apparent molecular mass of 97 kDa was immunoprecipitated from cell lysates with antiserum to ADP-ribosylated elongation factor 2 (EF-2). The {sup 32}P labeling of the protein was enhanced by culturing cells in medium containing 2% serum instead of 10% serum. The {sup 32}P label was completely removed from the protein by treatment with snake venom phosphodiesterase and the digestion product was identified as ({sup 32}P)AMP, indicating the protein was mono-ADP-ribosylated. HPLC analysis of tryptic peptides of the {sup 32}P-labeled 97-kDa protein and purified EF-2, which was ADP-ribosylated in vitro with diphtheria toxin fragment A and ({sup 32}P)NAD, demonstrated an identical labeled peptide in the two proteins. The data strongly suggest that EF-2 was endogenously ADP-ribosylated in pyBHK cells. Maximum incorporation of radioactivity in EF-2 occurred by 12 hr and remained constant over the subsequent 12 hr. It was estimated that 30-35% of the EF-2 was ADP-ribosylated in cells cultured in medium containing 2% serum. When {sup 32}P-labeled cultures were incubated in medium containing unlabeled phosphate, the {sup 32}P label was lost from the EF-2 within 30 min.

  16. Improving the quality of radiographic images acquired with conical radiation beams through divergence correction and filtering

    NASA Astrophysics Data System (ADS)

    Silvani, M. I.; Almeida, G. L.; Latini, R. M.; Bellido, A. V. B.; Souza, E. S.; Lopes, R. T.

    2015-07-01

    Earlier works have shown the feasibility to correct the deformation of the attenuation map in radiographs acquired with conical radiation beams provided that the inspected object could be expressed into analytical geometry terms. This correction reduces the contribution of the main object in the radiograph, allowing thus the visualization of its otherwise concealed heterogeneities. However, the non-punctual character of the source demanded a cumbersome trial-and-error approach in order to determine the proper correction parameters for the algorithm. Within this frame, this work addresses the improvement of radiographs of specially tailored test-objects acquired with a conical beam through correction of its divergence by using the information contained in the image itself. The corrected images have afterwards undergone a filtration in the frequency domain aiming at the reduction of statistical fluctuation and noise by using a 2D Fourier transform. All radiographs have been acquired using 165Dy and 198Au gamma-ray sources produced at the Argonauta research reactor in Institutode Engenharia Nuclear - CNEN, and an X-ray sensitive imaging plate as detector. The processed images exhibit features otherwise invisible in the original ones. Their processing by conventional histogram equalization carried out for comparison purposes did not succeed to detect those features.

  17. Semiphenomenological approximation of the sums of experimental radiative strength functions for dipole gamma transitions of energy E{sub {gamma}}below the neutron binding energy B{sub n} for mass numbers in the range 40 {<=} A {<=} 200

    SciTech Connect

    Sukhovoj, A. M. Furman, W. I. Khitrov, V. A.

    2008-06-15

    The sums of radiative strength functions for primary dipole gamma transitions, k(E1) + k(M1), are approximated to a high precision by a superposition of two functional dependences in the energy range 0.5 < E{sub 1} < B{sub n} - 0.5 MeV for the {sup 40}K, {sup 60}Co, {sup 71,74}Ge, {sup 80}Br, {sup 114}Cd, {sup 118}Sn, {sup 124,125}Te, {sup 128}I, {sup 137,138,139}Ba, {sup 140}La, {sup 150}Sm, {sup 156,158}Gd, {sup 160}Tb, {sup 163,164,165}Dy, {sup 166}Ho, {sup 168}Er, {sup 170}Tm, {sup 174}Yb, {sup 176,177}Lu, {sup 181}Hf, {sup 182}Ta, {sup 183,184,185,187}W, {sup 188,190,191,193}Os, {sup 192}Ir, {sup 196}Pt, {sup 198}Au, and {sup 200}Hg nuclei. It is shown that, in any nuclei, radiative strength functions are a dynamical quantity and that the values of k(E1) + k(M1) for specific energies of gamma transitions and specific nuclei are determined by the structure of decaying and excited levels, at least up to the neutron binding energy B{sub n}.

  18. Radiation transmission data for radionuclides and materials relevant to brachytherapy facility shielding.

    PubMed

    Papagiannis, P; Baltas, D; Granero, D; Pérez-Calatayud, J; Gimeno, J; Ballester, F; Venselaar, J L M

    2008-11-01

    To address the limited availability of radiation shielding data for brachytherapy as well as some disparity in existing data, Monte Carlo simulation was used to generate radiation transmission data for 60Co, 137CS, 198Au, 192Ir 169Yb, 170Tm, 131Cs, 125I, and 103pd photons through concrete, stainless steel, lead, as well as lead glass and baryte concrete. Results accounting for the oblique incidence of radiation to the barrier, spectral variation with barrier thickness, and broad beam conditions in a realistic geometry are compared to corresponding data in the literature in terms of the half value layer (HVL) and tenth value layer (TVL) indices. It is also shown that radiation shielding calculations using HVL or TVL values could overestimate or underestimate the barrier thickness required to achieve a certain reduction in radiation transmission. This questions the use of HVL or TVL indices instead of the actual transmission data. Therefore, a three-parameter model is fitted to results of this work to facilitate accurate and simple radiation shielding calculations.

  19. Thermal-neutron cross sections and resonance integrals of 138Ba and 141Pr using Am-Be neutron source

    NASA Astrophysics Data System (ADS)

    Panikkath, Priyada; Mohanakrishnan, P.

    2016-09-01

    The thermal-neutron capture cross sections and resonance integrals of 138Ba(n, γ)139Ba and 141Pr(n, γ)142Pr were measured by activation method using an isotopic Am-Be neutron source. The estimations were with respect to that of 55Mn(n, γ)56Mn and 197Au(n, γ)198Au reference monitors. The measured thermal-capture cross section of 138 Ba with respect to 55 Mn is 0.410±0.023 b and with respect to 197 Au is 0.386±0.019 b. The measured thermal-capture cross section of 141 Pr with respect to 55 Mn is 11.36±1.29 b and with respect to 197 Au is 10.43±1.14 b. The resonance integrals for 138 Ba are 0.380±0.033 b (55 Mn) and 0.364±0.027 b (197 Au) and for 141 Pr are 21.05±2.88 b (55 Mn) and 15.27±1.87 b (197 Au). The comparison between the present measurements and various reported values are discussed. The cross sections corresponding to the selected isotopes are measured using an Am-Be source facility for the first time.

  20. Measurements of neutron capture cross sections on 70Zn at 0.96 and 1.69 MeV

    NASA Astrophysics Data System (ADS)

    Punte, L. R. M.; Lalremruata, B.; Otuka, N.; Suryanarayana, S. V.; Iwamoto, Y.; Pachuau, Rebecca; Satheesh, B.; Thanga, H. H.; Danu, L. S.; Desai, V. V.; Hlondo, L. R.; Kailas, S.; Ganesan, S.; Nayak, B. K.; Saxena, A.

    2017-02-01

    The cross sections of the 70Zn(n ,γ )Zn71m (T1 /2=3.96 ±0.05 -h ) reaction have been measured relative to the 197Au(n ,γ )198Au cross sections at 0.96 and 1.69 MeV using a 7Li(p ,n )7Be neutron source and activation technique. The cross section of this reaction has been measured for the first time in the MeV region. The new experimental cross sections have been compared with the theoretical prediction by talys-1.6 with various level-density models and γ -ray strength functions as well as the tendl-2015 library. The talys-1.6 calculation with the generalized superfluid level-density model and Kopecky-Uhl generalized Lorentzian γ -ray strength function predicted the new experimental cross sections at both incident energies. The 70Zn(n ,γ ) g+m 71Zn total capture cross sections have also been derived by applying the evaluated isomeric ratios in the tendl-2015 library to the measured partial capture cross sections. The spectrum averaged total capture cross sections derived in the present paper agree well with the jendl-4.0 library at 0.96 MeV, whereas it lies between the tendl-2015 and the jendl-4.0 libraries at 1.69 MeV.

  1. Neutron capture cross-section studies of Tellurium isotopes for neutrinoless double beta decay applications

    NASA Astrophysics Data System (ADS)

    Bhike, Megha; Tornow, Werner

    2014-09-01

    The CUORE detector at Gran Sasso, aimed at searching for neutrinoless double-beta decay of 130Te, employs an array of TeO2 bolometer modules. To understand and identify the contribution of muon and (α,n) induced neutrons to the CUORE background, fast neutron cature cross-section data of the tellurium isotopes 126Te, 128Te and 130Te have been measured with the activation method at eight different energies in the neutron energy range 0.5-7.5 MeV. Plastic pill boxes of diameter 1.6 cm and width 1 cm containing Te were irradiated with mono-energetic neutrons produced via the 3H(p,n)3He and 2H(d,n)3He reactions. The cross-sections were determined relative to the 197Au(n, γ)198Au and 115In(n,n')115m In standard cross sections. The activities of the products were measured using 60% lead-shielded HPGe detectors at TUNL's low background counting facility. The present results are compared with the evaluated data from TENDL-2012, ENDF/B-VII.1, JEFF-3.2 and JENDL-4.0, as well as with literature data.

  2. Radiation transmission data for radionuclides and materials relevant to brachytherapy facility shielding

    SciTech Connect

    Papagiannis, P.; Baltas, D.; Granero, D.; Perez-Calatayud, J.; Gimeno, J.; Ballester, F.; Venselaar, J. L. M.

    2008-11-15

    To address the limited availability of radiation shielding data for brachytherapy as well as some disparity in existing data, Monte Carlo simulation was used to generate radiation transmission data for {sup 60}Co, {sup 137}Cs, {sup 198}Au, {sup 192}Ir, {sup 169}Yb, {sup 170}Tm, {sup 131}Cs, {sup 125}I, and {sup 103}Pd photons through concrete, stainless steel, lead, as well as lead glass and baryte concrete. Results accounting for the oblique incidence of radiation to the barrier, spectral variation with barrier thickness, and broad beam conditions in a realistic geometry are compared to corresponding data in the literature in terms of the half value layer (HVL) and tenth value layer (TVL) indices. It is also shown that radiation shielding calculations using HVL or TVL values could overestimate or underestimate the barrier thickness required to achieve a certain reduction in radiation transmission. This questions the use of HVL or TVL indices instead of the actual transmission data. Therefore, a three-parameter model is fitted to results of this work to facilitate accurate and simple radiation shielding calculations.

  3. [Bioaccumulation of silver and gold nanoparticles in organs and tissues of rats by neutron activation analysis].

    PubMed

    Buzulukov, Iu P; Arianova, E A; Demin, V F; Safenkova, I V; Gmoshinskiĭ, I V; Tutel'ian, V A

    2014-01-01

    Bioaccumulation of silver (Ag) and gold (Au) nanoparticles (NPs) with mean sizes of 6 nm and 35 nm, respectively, has been studied after their intragastric administration to rats at a dose of 100 μg/kg of body weight for 28 or 14 days. The organs and tissues (liver, kidney, spleen, heart, gonads brain, and blood) were subjected to thermal neutron activation, and, then, the activity of the 110mAg and 198Au isotopes generated was measured. The NPs of both metals were detected in all biological samples studied, the highest specific weight and content of Ag NP being found in the liver, and those of Au being found in kidneys of animals. The content of Ag NPs detected in the brain was 66.4 ± 5.6 ng (36 ng/g tissue), no more than 7% ofthese NPs being localized in the lumen of brain blood vessels. The content of Ag and Au NPs found in organs and tissues of rats could be regarded as nonhazardous (nontoxic) in accordance with the known literature data.

  4. Do Radioactive Half-Lives Depend on the Earth-Sun Distance?

    NASA Astrophysics Data System (ADS)

    Goodwin, John; Hardy, John; Iacob, Victor; Golovko, Victor

    2010-11-01

    In recent articles [1-4], Jenkins et al. claim to have evidence that radioactive half-lives vary as a function of the earth-to-sun distance. They base their claims on data obtained by others over the space of several years -- the decay of ^32Si as measured at Brookhaven National Laboratory [5] from 1982-85 and that of ^226Ra as measured at the Physikalisch Technische Bundesanstalt in Germany [6] from 1984-88 -- in which the decay rates show a small but statistically significant oscillation with a period of one year and approximately correlated with the earth-sun distance (and with the seasons). Here we report a series of seven measurements of the ^198Au half-life (see [7] for a description of two of these) made by us at various intervals over a period of one aphelion-aphelion cycle. Each measured half-life has a precision of about 0.02%. There is no evidence of any deviation from a constant half-life. [1] J. H. Jenkins et al., Astropart. Phys. 31 407 (2009) [2] J. H. Jenkins et al., Astropart. Phys. 32, 42 (2010) [3] E. Fischbach et al., Space Sci. Rev. 145, 285 (2009) [4] J. Jenkins et al., arxiv:0912:5385v1 [5] D. Alburger et al., Earth and Planet. Sci. Lett. 78, 168 (1986) [6] H. Siegert et al., Appl. Radiat. Isot. 49, 1397 (1998) [7] J. R. Goodwin et al., Eur. Phys. J. A 34, 271 (2007).

  5. The measurement of skin lymph flow by isotope clearance--reliability, reproducibility, injection dynamics, and the effect of massage

    SciTech Connect

    Mortimer, P.S.; Simmonds, R.; Rezvani, M.; Robbins, M.; Hopewell, J.W.; Ryan, T.J. )

    1990-12-01

    The measurement of skin lymph flow was investigated using an isotope clearance technique (ICT). Multiple lymph flow determinations were undertaken in the skin of anaesthetized large white pigs to test for reproducibility, ascertain the most suitable tracer, study the influence of injection dynamics, and observe the effect of massage as a stimulus to lymph flow. Blood clearance of tracer was also investigated. Results demonstrated that lymphatic clearance is a monoexponential function with good reproducibility under controlled laboratory conditions. 99mTc-colloid (TCK17 Cis) compared favorably with 131I-human serum albumin as a tracer and both performed better than colloid gold (198Au). Lymph flow was significantly faster in one pig than in the other. No difference existed between left and right sides or between caudal and rostral sites on each flank, but clearance was significantly slower in thigh than flank skin. Sub-epidermal injections cleared faster and more consistently than either deep or subcutaneous injections. Neither injection volume nor needle tract backflow of tracer influenced results, but local massage significantly enhanced clearance. Escape of 99mTc-colloid by the blood was negligible. These results indicate that skin lymph flow can be reliably measured when conditions are controlled. Extrinsic factors such as massage strongly influence lymph flow. Greater sensitivity in detecting degrees of lymphatic insufficiency may be achieved if a standardized stimulus to lymph flow is administered during isotope clearance measurement.

  6. Interstitial brachytherapy for pancreatic cancer: Report of seven cases treated with 125I and a review of the literature

    SciTech Connect

    Montemaggi, P.; Dobelbower, R.; Crucitti, F.; Caracciolo, F.; Morganti, A.G.; Smaniotto, D.; Luzi, S.; Cellini, N. )

    1991-07-01

    Since 1975, seven groups of investigators have reported clinical results of interstitial brachytherapy (IBT) for pancreatic cancer. The reports are comprised of data from 254 patients, 21 of whom died in the postoperative period for an overall operative mortality rate of 8.7%. Operative mortality rate range from 0% to 32% in individual reports. Most patients have been treated with 125I, although 25 patients were treated with 198Au seeds. Most investigators report combining IBT with external beam radiation therapy (EBRT) {plus minus} adjuvant chemotherapy. In general, IBT has been associated with considerable morbidity. Median patient survival time has not exceeded 15 months. This report describes an additional seven patients with locally unresectable pancreatic cancer, without distant metastases, treated primarily with 60 to 100 Gy matched peripheral dose (MPD) by 125I IBT. One patient died postoperatively of a pulmonary embolus. Four of the remaining six patients were also treated with modest doses (10.5 to 30 Gy) of EBRT late in the course of the disease for local tumor progression. One developed a pancreaticocutaneous fistula, and one developed exacerbation of pre-existing diabetes mellitus. The median patient survival time from the date of IBT was 7 months (range: 0 to 21 months). One patient is alive without clinical evidence of cancer 9 months after IBT. 25 refs.

  7. Image-based dosimetry of an implanted radioactive stent using intravascular ultrasound

    NASA Astrophysics Data System (ADS)

    Peterson, Stephen W.

    Angioplasty has become an increasingly popular and effective treatment for heart disease. Unfortunately, restenosis, a cellular and biological reaction to the procedure, has hindered its effectiveness. Two of the most successful methods of inhibiting restenosis are radiation and stents. The combination of these two components, radioactive stents, is not as common as some of the other methods, yet still has potential of slowing restenosis. Investigation into source characteristics and artery wall radiobiology may illuminate some possible solutions to the problems of restenosis. This work has developed a calculational method to look at in-vivo images of implanted stents and determine the dose to the artery walls in order to test different source characteristics. The images are Intravascular Ultrasound (IVUS) cross-sectional slices of the stent and the artery. From these images, it is possible to determine the implanted stent structure. The pieces of the stent are identified in the images and modeled in a Monte Carlo simulation, using MCNP4c3. The simulation results were combined with the images to give three-dimensional absolute dose contours of the stent. The absolute dose values were verified using radiochromic film and 198Au-plated stents. This work was able to successfully verify the dose results and create a three-dimensional dose map of the implanted stent.

  8. Semiphenomenological approximation of the sums of experimental radiative strength functions for dipole gamma transitions of energy E γ below the neutron binding energy B n for mass numbers in the range 40 ≤ A ≤ 200

    NASA Astrophysics Data System (ADS)

    Sukhovoj, A. M.; Furman, W. I.; Khitrov, V. A.

    2008-06-01

    The sums of radiative strength functions for primary dipole gamma transitions, k( E1) + k( M1), are approximated to a high precision by a superposition of two functional dependences in the energy range 0.5 < E 1 < B n - 0.5 MeV for the 40K, 60Co, 71,74Ge, 80Br, 114Cd, 118Sn, 124,125Te, 128I, 137,138,139Ba, 140La, 150Sm, 156,158Gd, 160Tb, 163,164,165Dy, 166Ho, 168Er, 170Tm, 174Yb, 176,177Lu, 181Hf, 182Ta, 183,184,185,187W, 188,190,191,193Os, 192Ir, 196Pt, 198Au, and 200Hg nuclei. It is shown that, in any nuclei, radiative strength functions are a dynamical quantity and that the values of k( E1) + k( M1) for specific energies of gamma transitions and specific nuclei are determined by the structure of decaying and excited levels, at least up to the neutron binding energy B n .

  9. [Studies on the lymph drainage of the eye. 1. Quantitative registration of the lymph drainage from the orbita of the rabbit with radioactive tracers (author's transl)].

    PubMed

    Grüntzig, J; Schicha, H; Kiem, J; Becker, V; Feinendegen, L E

    1977-05-01

    For the investigation on the lymph drainage from the orbita 99mTc-microcolloid, 99mTc-albumin and 198Au-colloid have been injected into the retrobulbar space of 24 rabbits. Measurements of the activity's distribution have been made in vivo with an Anger type camera (pho-Gamma-IV Hp, Searle Nuclear Chicago) and in vitro after section with a sodium iodine crystal well counter (Clinimat-200, Picker). A significant concentration of the activity could be observed for the most part in the equilateral Lymphonoduli cervicales profundi and superficiales and for the less part also in the equilateral Lymphonoduli mandibulares and contralateral Lymphonoduli cervicales profoundi. Furthermore significant activities could be pointed out in the optic nerves as well as in the contralateral retrobulbar space. The data substantiate a lymph drainage from the orbita as well as a partly reverse "prelymphatic-lymphatic" flow and are of importance for discussions about lymphostatic ophthalmopathy and sympathetic ophthalmitis. For application on man 99mTc-microcolloid (Lymphoscint) shoud be preferred, because with this tracer 1. the lymph drainage can be measured quantitatively quite exactly by extern measurement with the Anger-camera, 2. the allowable radiation dosis for the lens is lying between 0.5-1.0 rad/mCi.

  10. Validation of the MCNP computational model for neutron flux distribution with the neutron activation analysis measurement

    NASA Astrophysics Data System (ADS)

    Tiyapun, K.; Chimtin, M.; Munsorn, S.; Somchit, S.

    2015-05-01

    The objective of this work is to demonstrate the method for validating the predication of the calculation methods for neutron flux distribution in the irradiation tubes of TRIGA research reactor (TRR-1/M1) using the MCNP computer code model. The reaction rate using in the experiment includes 27Al(n, α)24Na and 197Au(n, γ)198Au reactions. Aluminium (99.9 wt%) and gold (0.1 wt%) foils and the gold foils covered with cadmium were irradiated in 9 locations in the core referred to as CT, C8, C12, F3, F12, F22, F29, G5, and G33. The experimental results were compared to the calculations performed using MCNP which consisted of the detailed geometrical model of the reactor core. The results from the experimental and calculated normalized reaction rates in the reactor core are in good agreement for both reactions showing that the material and geometrical properties of the reactor core are modelled very well. The results indicated that the difference between the experimental measurements and the calculation of the reactor core using the MCNP geometrical model was below 10%. In conclusion the MCNP computational model which was used to calculate the neutron flux and reaction rate distribution in the reactor core can be used for others reactor core parameters including neutron spectra calculation, dose rate calculation, power peaking factors calculation and optimization of research reactor utilization in the future with the confidence in the accuracy and reliability of the calculation.

  11. Distribution of total radiation widths for neutron resonances of Pt isotopes

    NASA Astrophysics Data System (ADS)

    Koehler, P. E.; Bečvář, F.; Krtička, M.

    2015-05-01

    High quality neutron capture and transmission data were measured on isotopically enriched 192,194,195,196Pt and natural Pt samples at ORELA. R-matrix analysis of this data revealed resonance parameters for 159, 413, 423, 258, and 11 neutron resonances for neutron energies below 5.0, 16.0, 7.5, 16.0, and 5.0 keV for 192,194,195,196,198Pt+n, respectively. Earlier analysis of data on reduced neutron widths, Γ0n, showed that the distributions of Γ0n for 192,194Pt deviate significantly from the Porter-Thomas distribution (PTD) predicted by random matrix theory. In this contribution we report on preliminary results of the analysis of distribution of total radiation widths, Γγ, in 192,194,195,196Pt+n reactions. Comparison of experimental data with predictions made within the nuclear statistical model indicates that standard models of Photon Strength Functions (PSFs) and Nuclear Level Density predict Γγ distributions which are too narrow. We found that satisfactory agreement between experimental and simulated distributions can be obtained only by a strong suppression of the PSFs at low γ-ray energies and/or by violation of the usual assumption that primary transitions from neutron resonances follow the PTD. The shape of PSFs needed for reproduction of our Γγ data also nicely reproduces spectra from several (n,γ) experiments on the neighbor nuclide 198Au.

  12. Plan for radionuclide tracer studies of the residence time distribution in the Wilsonville dissolver and preheater

    SciTech Connect

    Jolley, R.L.; Begovich, J.M.; Brashear, H.R.; Case, N.; Clark, T.G.; Emery, J.F.; Patton, B.D.; Rodgers, B.R.; Villiers-Fisher, J.F.; Watson, J.S.

    1983-12-01

    Stimulus-response measurements using radiotracers to measure residence time distribution (RTD) and hydrodynamic parameters for the preheaters and dissolvers at the Ft. Lewis Solvent Refined Coal (SRC) and the Exxon Donor Solvent (EDS) coal conversion pilot plants are reviewed. A plan is also presented for a series of radioactive tracer studies proposed for the Advanced Coal Liquefaction Facility at Wilsonville, Alabama, to measure the RTD for the preheater and dissolvers in the SRC-I mode. The tracer for the gas phase will be /sup 133/Xe, and /sup 198/Au (on carbonized resin or as an aqueous colloidal suspension) will be used as the slurry tracer. Four experimental phases are recommended for the RTD tracer studies: (1) preheater; (2) dissolver with 100% takeoff; (3) dissolver with 100% takeoff and solids withdrawal; and (4) dissolver with 50% takeoff. Eighteen gas-tracer and 22 liquid-tracer injections are projected to accomplish the four experimental phases. Two to four tracer injections are projected for preliminary tests to ensure the capability of safe injection of the radiotracers and the collection of statistically significant data. A complete projected cost and time schedule is provided, including procurement of necessary components, preparation of the radiotracers, assembly and testing of tracer injection apparatus and detection systems, onsite work and tracer injections, laboratory experimentation, data analysis, and report writing.

  13. Effect of exercise on redistribution and clearance of inhaled particles from hamster lungs

    SciTech Connect

    Sweeney, T.D.; Tryka, A.F.; Brain, J.D. )

    1990-03-01

    Does exercise alter the redistribution and clearance of particles from the lungs Sedentary hamsters and hamsters that were exercise trained by voluntary wheel running for the previous 5 wk were exposed to a 198Au-labeled aerosol for 25 min. Six trained and 6 sedentary animals were killed within 5 min after the exposure (day 0); the same number were killed 5 days later. The trained hamsters ran ad libitum during those 5 days. The lungs of all animals were excised, dried at total lung capacity, sliced into 1-mm-thick sections, and dissected into pieces that were counted for radioactivity and weighed. On day 0, trained hamsters had 80% more particles per milligram of lung than sedentary hamsters, although both were exposed under identical conditions of restraint. After five days, exercising hamsters cleared 38% of the particles present at day 0, whereas sedentary animals removed only 15%. Significant clearance was observed from the middle lung regions of sedentary hamsters and from all lung regions in exercising hamsters. We conclude that exercise can enhance the redistribution and clearance of particles from the lungs; the mechanisms responsible are as yet unclear.

  14. Pharmacokinetics and distribution of dietary tributyltin and methylmercury in the snow crab (Chionoecetes opilio)

    SciTech Connect

    Rouleau, C.; Gobeil, C.; Tjaelve, H.

    1999-10-01

    The pharmacokinetics and distribution of a single 5-{micro}g dietary dose of radiolabeled [{sup 113}Sn]tributyltin (TBT) and [{sup 203}Hg]methylmercury (MeHg) were studied over 154 days in the snow crab, using in vivo gamma counting and whole-body autoradiography. Experiment was done under conditions typical of those encountered in the cold natural habitat of this crustacean. Retention efficiency was high for both compounds, and two kinetic pools could be distinguished. Elimination of the first pool proceeded within 20--80 days, but it accounted for 27--62% of the assimilated TBT, compared to 8--11% for MeHg. Biological half-life of the second pool was 33--187 days for TBT and 520--650 days for MeHg. Autoradiographic and dissection data revealed a less homogeneous distribution of the radiolabel and much higher radioactivity in gut lumen for TBT compared to MeHg. This suggests that the larger size of the first pool in the case of TBT resulted from metabolization in the hepatopancreas and fecal elimination of the metabolites. The whole-body biomagnification factor (BMF) that would result from the long-term chronic exposure of snow crab to TBT-contaminated food was estimated as 0.1--0.6. Although these BMF values were an order of magnitude lower than those estimated for MeHg, they are not negligible and indicate that uptake of TBT via food may be an important accumulation route.

  15. Accumulation of waterborne mercury(II) in specific areas of fish brain

    SciTech Connect

    Rouleau, C.; Borg-Neczak, K.; Gottofrey, J.; Tjaelve, H.

    1999-10-01

    The authors used whole-body autoradiography to study the distribution of {sup 203}Hg(II) in the central nervous system of brown (Salmo trutta) and rainbow (Oncorhynchus mykiss) trout. Fish were either exposed to waterborne Hg(II) for 7 and 21 d or they received an intravenous injection of the metal and were sacrificed 1 and 21 d later. Mercury did not accumulate in the brain after intravenous injection, indicating that the blood-brain barrier is impervious to Hg in plasma. In contrast, Hg was accumulated in specific areas of the grain and spinal cord following water exposure. The specificity of the accumulation sites strongly suggests that waterborne Hg was taken up by water-exposed receptor cells of sensory nerves and subsequently transferred toward the brain by axonal transport, a normal physiological process for the transport of organelles and dissolved neuronal constituents along nerve axons. Accumulation of Hg in ventral horn ganglis is probably the result of leaching of metal from blood into muscle followed by uptake in motor plates. Axonal transport allows waterborne inorganic Hg, and possibly other xenobiotics, to circumvent the blood-brain barrier. Considering the importance of complex behavior in the life of fish, and the well-known deleterious effects of mercury on the nervous system, the toxicological significance of this uptake route needs to be assessed.

  16. Distribution kinetics of dietary methylmercury in the arctic charr (Salvelinus alpinus)

    SciTech Connect

    Ribeiro, C.A.O.; Rouleau, C.; Pelletier, E.; Audet, C.; Tjaelve, H.

    1999-03-15

    The authors fed immature 1+ arctic charr with a single dose of methyl[{sup 203}Hg]mercury (MeHg) and quantified distribution kinetics with a new and simple three-compartment caternary model having well-perfused viscera and blood as the central compartment (VB), whereas gut (G) and the rest of body (R) constituted the peripheral compartments. The model accurately described distribution kinetics of MeHg in the fish, using either data of MeHg content in compartments or blood concentration data. Despite the known fast translocation of MeHg between binding sites at the molecular level, its translocation rate between compartments was surprisingly slow, 27 days being needed to complete 95% of the transfer from gut to blood and 48 days for the subsequent transfer to compartment R. This probably results from a limitation of the stepwise transfer rate of MeHg from red blood cells, which contain most of blood MeHg, to plasma and then to tissues due to low plasmatic concentration of small mobile sulfhydryl ligands. The model presented is a convenient tool that could be used to compare the fate of MeHg and other organometals, such as butyltins and alkylleads, in various aquatic and terrestrial animal species.

  17. Influence of cations on the blue to purple transition of bacteriorhodopsin. Comparison of Ca2+ and Hg2+ binding and their effect on the surface potential.

    PubMed

    Duñach, M; Seigneuret, M; Rigaud, J L; Padrós, E

    1988-11-25

    We have investigated the effect of Ca2+ and Hg2+ binding on various properties of the blue membrane prepared by deionization of the Halobacterium halobium purple membrane. Binding of radioactive 45Ca2+ and 203Hg2+ was monitored by a filtration technique. Five high and medium affinity sites for Ca2+ and seven low affinity sites for Hg2+ were found per bacteriorhodopsin. Competitive binding was observed only for three Ca2+ and three Hg2+. Visible absorption studies indicated that Ca2+ binding could restore the purple color of bacteriorhodopsin while Hg2+ was inefficient. Hg2- could partially reverse to blue the Ca2+-regenerated purple membrane in parallel with the displacement of three Ca2+. Effects of cation binding on the surface potential of the membrane were measured by Electron Spin Resonance spectroscopy using a cationic spin-labeled amphiphile. Cations such as La3+, Ca2+, Mg2+, or Na+ strongly increased (i.e. rendered less negative) the surface potential. An univocal correlation was found between the cation-induced variation of surface potential and the extent of regeneration of the purple color. Hg2+ induced a smaller increase in surface potential than that corresponding to the effective divalent cations. This lower effect appears to be due to binding to sites not related to those of other cations.

  18. Measurement of activation cross-sections for high-energy neutron-induced reactions of Bi and Pb

    NASA Astrophysics Data System (ADS)

    Zaman, Muhammad; Kim, Guinyun; Kim, Kwangsoo; Naik, Haladhara; Shahid, Muhammad; Lee, Manwoo

    2015-08-01

    The cross-sections for 209Bi(n, 4n)206Bi, 209Bi(n, 5n)205Bi, natPb(n, xn)204mPb, natPb(n, xn)203Pb, natPb(n, xn)202mPb,natPb(n, xn)201Pb, natPb(n, xn)200Pb, natPb(n, αxn)203Hg and natPb(n, p xn)202Tl reactions were determined at the Korean Institute of Radiological and Medical Sciences (KIRAMS), Korea in the neutron energy range of 15.2 to 37.2 MeV. The above cross-sections were obtained by using the activation and off-line γ-ray spectrometric technique. The quasi-monoenergetic neutron used for the above reactions are based on the 9Be(p, n) reaction. Simulations of the spectral flux from the Be target were done using the MCNPX program. The cross-sections were estimated with the TALYS 1.6 code using the default parameter. The data from the present work and literature were compared with the data from the EAF-2010 and the TENDL-2013 libraries, and calculated values of TALYS 1.6 code. It shows that appropriate level density model, the γ-ray strength function, and the spin cut-off parameter are needed to obtain a good agreement between experimental data and theoretical values from TALYS 1.6 code.

  19. Simultaneous radioassays of bacterial production and mercury methylation in the periphyton of a tropical and a temperate wetland.

    PubMed

    Guimarães, J R D; Mauro, J B N; Meili, M; Sundbom, M; Haglund, A L; Coelho-Souza, S A; Hylander, L D

    2006-10-01

    Laboratory radioassays were made to study mercury (Hg) methylation together with bacterial production in the periphyton of two aquatic macrophytes, the submerged Myriophyllum spicatum, from a constructed wetland in Sweden and the floating Eichhornia crassipes, from a eutrophied tropical lake in Brazil. Time course incubations were made by addition of (203)HgCl(2) and the methylmercury formed was extracted at pre-defined time intervals. Bacterial production ((14)C-leucine incorporation) was measured at the same time intervals, with plants removed from parallel incubations made with and without addition of cold HgCl(2). For E. crassipes, higher methylmercury production was observed at elevated bacterial production, whereas for M. spicatum, the bacterial production was significantly lower, and Hg methylation was below the detection limit. The combined results confirm the importance of microbial processes for Hg methylation, although other factors are known to influence this process in complex ways. The addition of Hg did not significantly influence bacterial production, while the incubation temperatures used (25 and 35 degrees C) resulted in different methylation rates. Radiotracer techniques for measurements of bacterial production such as (14)C-leucine uptake can provide useful insights into the Hg cycle in aquatic environments, and our data suggest that they may be used as a proxy of mercury methylation potentials.

  20. Selective solubilization of membrane proteins differentially labeled by p-chloromercuribenzenesulfonic acid in the presence of sucrose

    SciTech Connect

    M'Batchi, B.; Pichelin, D.; Delrot, S.

    1987-03-01

    Broadbean (Vicia faba L.) leaf discs have been incubated with the slowly permeant thiol reagent (/sup 203/Hg)-para-chloromercuribenzenesulfonic acid (PCMBS) in the presence or in the absence of sucrose, and the release of PCMBS-labeled proteins has been monitored in media containing various concentrations of urea, ethylene glycol-bis-(..beta..-aminoethyl ether)-N, N, N', N'-tetraacetic acid (EGTA), sodium cholate, sodium dodecyl sulfate, Triton X-100, octylglucoside or (3-(3-cholamidopropyl)-dimethylammonio) 1-propane-sulfonate)(CHAPS). The proteins differentially labeled by PCMBS in the presence of sucrose which, on the basis of previous results, are assumed to included the sucrose carrier, were preferentially solubilized by 1% CHAPS, 1% octylglucoside, or 1% Triton X-100. Other PCMBS-labeled proteins (background proteins) could be partially removed by EGTA, urea, or 0.1% cholate. Sequential treatment by 10 mM EGTA and 1% CHAPS was found to give a fraction highly enriched in the differentially labeled proteins. Analysis of the specific activity of microsomal pellets suggests that the results obtained with leaf discs give a good account of what is occurring at the plasma membrane level. These data, which suggest that the proteins differentially labeled, by PCMBS in the presence of sucrose are intrinsic membrane proteins, can be used to solubilize these proteins from microsomal fractions.

  1. Mercury distribution, methylation and volatilization in microcosms with and without the sea anemone Bunodosoma caissarum.

    PubMed

    Rizzini Ansari, Nafisa; Correia, Raquel Rose Silva; Fernandez, Marcos Antônio; Cordeiro, Renato Campello; Guimarães, Jean Remy Davée

    2015-03-15

    Mercury (Hg) has a complex biogeochemical cycle in aquatic environments. Its most toxic form, methylmercury (MeHg), is produced by microorganisms. This study investigated how the sea anemone Bunodosoma caissarum affects Hg distribution, methylation and volatilization in laboratory model systems. (203)Hg was added to microcosms and its distribution in seawater, specimens and air was periodically measured by gamma spectrometry. MeHg was measured by liquid scintillation. After the uptake period, specimens had a bioconcentration factor of 70 and in microcosms with and without B. caissarum, respectively 0.05% and 0.32% of the initial spike was found as MeHg. After depuration, MeHg in specimens ranged from 0.2% to 2.4% of total Hg. Microcosms with B. caissarum had higher Hg volatilization (58%) than controls (17%), possibly due to Hg(2+) reduction mediated by microorganisms associated with its tissues and mucus secretions. Marine organisms and their associated microbiota may play a role in Hg and MeHg cycling. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. A study of gamma attenuation parameters in poly methyl methacrylate and Kapton

    NASA Astrophysics Data System (ADS)

    Manjunatha, H. C.

    2017-08-01

    Poly methyl methacrylate (PMMA) and Kapton polyimide are polymers used for various aerospace applications. We have measured the gamma attenuation parameters such as mass attenuation coefficient, effective atomic number and electron density of PMMA and Kapton polyimide for various gamma sources of energy ranging from 84 keV to 1330 keV (170Tm, 57Co, 141Ce, 203Hg, 51Cr, 113Sn, 22Na, 137Cs, 60Co, 22Na and 60Co). The measured values agree with the theoretical values. In the present work, we have also computed energy absorption build-up factors and exposure buildup factor of PMMA and Kapton polyimide for wide energy range (0.015-15 MeV) up to the penetration depth of 40 mean free path using Geometrical Progression fitting method. The values of these parameters have been found to change with energy and interaction of gamma with the medium. The present study on gamma attenuation parameters are expected to be helpful in dosimetry, radiation shielding and other radiation physics based applications. The experimental data on the mass attenuation coefficients for Kapton and PMMA is not available in literature. To my knowledge data available e.g. in the NIST data base are based on extrapolations from the measurement of mass attenuation coefficients for the elements. Hence this work is first of its kind and it is useful in the various field of Polymers.

  3. Initial uptake and insulin releasing action of chloromercuribenzene-p-sulphonic acid (CMBS) in suspensions of pancreatic islet cells.

    PubMed

    Idahl, L A; Lernmark, A; Söderberg, M; Winblad, B

    1980-04-01

    The effects of chloromercuribenzene-p-sulphonic acid on dispersed cells prepared from beta-cell-rich ob/ob-mouse islets were studied. 1) Chloromercuribenzene-p-sulphonic acid at concentrations of 0.1 mmol/l or higher diminished cell viability which was partially counteracted by increasing concentrations of bovine serum albumin. 2) The uptake of 203Hg-chloromercuribenzene-p-sulphonic acid after incubation for 4 seconds or longer showed that most of the non-toxic concentrations of chloromercuribenzene-p-sulphonic acid was bound to the cell within 40 seconds. Maximal uptake was achieved after 3 minutes of incubation. The uptake of radioactive chloromercuribenzene-p-sulphonic acid was inhibited by bovine serum albumin. 3) The dynamics of insulin release from perifused dispersed beta-cells embedded in fibrin showed a maximal 40--50-fold stimulation by 0.03 mmol/l chloromercuribenzene-p-sulphonic acid within 10 minutes of perifusion. 4) Scanning electron microscopy of beta-cells revealed no major changes in the cell surface under conditions of maximal binding and insulin releasing effects of chloromercuribenzene-p-sulphonic acid. These data support the concept that the ability of chloromercuribenzene-p-sulphonic acid to induce insulin release is related to its initial binding to the beta-cell surface. The binding of chloromercuribenzene-p-sulphonic acid and the subsequent release of insulin seem to occur without major changes in beta-cell surface morphology.

  4. Normal operation and maintenance safety lessons from the ITER US PbLi test blanket module program for a US FNSF and DEMO

    SciTech Connect

    L. C. Cadwallader; C. P. C. Wong; M. Abdou; B. B. Morely; B.J Merrill

    2014-10-01

    A leading power reactor breeding blanket candidate for a fusion demonstration power plant (DEMO) being pursued by the US Fusion Community is the Dual Coolant Lead Lithium (DCLL) concept. The safety hazards associated with the DCLL concept as a reactor blanket have been examined in several US design studies. These studies identify the largest radiological hazards as those associated with the dust generation by plasma erosion of plasma blanket module first walls, oxidation of blanket structures at high temperature in air or steam, inventories of tritium bred in or permeating through the ferritic steel structures of the blanket module and blanket support systems, and the 210Po and 203Hg produced in the PbLi breeder/coolant. What these studies lack is the scrutiny associated with a licensing review of the DCLL concept. An insight into this process was gained during the US participation in the International Thermonuclear Experimental Reactor (ITER) Test Blanket Module (TBM) Program. In this paper we discuss the lessons learned during this activity and make safety proposals for the design of a Fusion Nuclear Science Facility (FNSF) or a DEMO that employs a lead lithium breeding blanket.

  5. Phosphorylation of the C proteins in heterogeneous ribonucleoprotein (hnRNP) particles in HeLa cells: Characterization of in vivo phosphorylation, comparison with in vitro phosphorylation using casein kinase II, and preliminary studies on the effects of phosphorylation on particle structure

    SciTech Connect

    Kleiman, N.J.

    1989-01-01

    Newly formed pre-messenger RNA associates with protein to form heterogeneous ribonucleoprotein (hnRNP) particles. In HeLa cells, hnRNP particles contain six core proteins. Two proteins, termed C{sub 1} and C{sub 2}, are phosphorylated in vitro by casein kinase 11 (CKII). C{sub 1} protein became {sup 32}P-labeled after HeLa cells were incubated with ({sup 32}P)-orthophosphate in vivo (ibid). Because phosphorylation is a ubiquitous regulatory mechanism, C protein phosphorylation was studied in greater detail. C protein phosphorylation in hnRNP particles was investigated in HeLa cells incubated with ({sup 32}P)-orthophosphate in vivo. Immunoblotting in pH 3.5-10 isoelectric focusing (IEF) gels indicated that C proteins focus only at pH 5.0. In pH 4.5-5.5 IEF gels, individually purified C, and 2 proteins resolve into the same four closely spaced, {sup 32}P-labeled bands. A fifth, unlabeled, more basic species was detached when hnRNP particles were purified without NaF. All {sup 32}P-labeled species contained identical amounts of {sup 32}P per unit protein suggesting that charge heterogeneity is not due to differential phosphorylation. Attempts to detect bound carbohydrate were unsuccessful. {sup 32}P-labeled phosphate was readily removed by potato acid phosphatase. E. coli alkaline phosphatase and snake venom phosphodiesterase were ineffective. {sup 32}P-label was found exclusively in phosphoserine. One-dimensional peptide mapping with chymotrypsin and S. aureus protease detected two phosphorylated peptides. C protein phosphorylation was also investigated in vitro. Incubation of hnRNP particles with rabbit liver CKII and {sup 32}P-ATP followed by IEF in pH 4.5-5.5 gels indicated that all four C protein species were {sup 32}P-labeled. {sup 32}P-label was found exclusively in phosphoserine.

  6. Inhibitory effect of endostatin gene therapy combined with phosphorus-32 colloid on tumour growth in Wistar rats.

    PubMed

    Gao, Huiqi; Zhu, Jing; Li, Yong; Fu, Peng; Shen, Baozhong

    2016-07-01

    Eighty healthy male Wistar rats, aged 5 weeks, weighing 100-120 g, were utilized for establishing tumour-bearing models by immediate Walker-256 cancerous ascites injection and randomly divided to four groups (n=20) treated with 0.2 ml solution containing saline, (32)P-colloid (0.3 mCi), endostatin gene (20 μg), endostatin gene combined with colloid (32)P. The effect of endostatin combined with a small dose of (32)P-colloidal on tumour growth in vivo was evaluated and the potential mechanism underlying the combined therapy was explored. We found that (32)P-colloid combined with endostatin exhibited higher inhibitory effect upon tumour growth compared with application of (32)P-colloid or endostatin alone, although three therapies all significantly inhibited tumour growth compared with saline control group. The higher inhibitory effect of (32)P-colloid combined with endostatin upon tumour growth might be attributed to a synergistic effect of inhibiting angiogenesis by endostatin and inducing apoptosis by (32)P-colloid, as demonstrated by microvessel density (MVD) and apoptotic index (AI) measurement. Combined therapy of (32)P-colloid and endostatin probably serves as a novel and efficacious therapy of tumour growth.

  7. Long Distance Transport in Macrocystis integrifolia

    PubMed Central

    Schmitz, Klaus; Srivastava, Lalit M.

    1979-01-01

    Discs from mature regions of Macrocystis blades picked up significantly more [32P]phosphate from the ambient medium than similar discs from young meristematic regions, and this uptake was higher in light than in darkness. Double-labeling experiments with NaH14CO3 and [32P]phosphate, using intact fronds as well as cut frond segments, indicated that 32P was translocated from mature blades to sink regions at velocities of 25 to 45 centimeters per hour, velocities comparable to 14C translocation velocity in the same material. There was a slight delay in transport of 32P which may be due to a delay in loading or to a high metabolism of 32P in the transporting channels. Histoautoradiography of stipe segments in the translocation pathway indicated that transport of label occurred in the peripheral parts of medulla. An analysis of 32P-labeled compounds in the fed blade and in the sieve tube sap, collected from basal cut ends of stipes, indicated major differences in labeling patterns. In the blade, a high proportion of 32P was recovered as inorganic phosphate and relatively small amounts were found in hexose mono- and diphosphates, UDPG and ATP. In the sieve tube sap, however, only a small amount of 32P was present as inorganic phosphate, a large proportion was found in hexose mono- and diphosphates, and appreciable amounts were present in ATP and UDPG. Images PMID:16660847

  8. Identification of the 64 kilodalton chloroplast stromal phosphoprotein as phosphoglucomutase. [Pisum sativum

    SciTech Connect

    Salvucci, M.E.; Drake, R.R.; Broadbent, K.P.; Haley, B.E. ); Hanson, K.R.; McHale, N.A. )

    1990-05-01

    Phosphorylation of the 64 kilodalton stromal phosphoprotein by incubation of pea (Pisum sativum) chloroplast extracts with ({gamma}-{sup 32}P)ATP decreased in the presence of Glc-6-P and Glc-1,6-P{sub 2}, but was stimulated by glucose. Two-dimensional gel electrophoresis following incubation of intact chloroplasts and stromal extracts with ({gamma}-{sup 32}P)ATP, or incubation of stromal extracts and partially purified phosphoglucomutase (EC 2.7.5.1) with ({sup 32}P)Glc-1-P showed that the identical 64 kilodalton polypeptide was labeled. A 62 kilodalton polypeptide was phosphorylated by incubation of tobacco (Nicotiana sylvestris) stromal extracts with either ({gamma}-{sup 32}P)ATP or ({sup 32}P)Glc-1-P. In contrast, an analogous polypeptide was not phosphorylated in extracts from a tobacco mutant deficient in plastid phosphoglucomutase activity. The results indicate that the 64 (or 62) kilodalton chloroplast stromal phosphoprotein is phosphoglucomutase.

  9. The effects of barbiturates on the metabolism of phosphatidic acid and phosphatidylinositol in rat brain synaptosomes.

    PubMed Central

    Miller, J C; Leung, I

    1979-01-01

    Barbiturates and diphenylhydantoin inhibit the carbamoylcholine-stimulated increase in 32P incorporation into phosphatidylinositol and phosphatidic acid, but have a relatively slight effect on the incorporation of 32P into these lipids in the absence of carbamoylcholine and no effect on 32P incorporation into phosphatidylcholine and phosphatidylethanolamine. Inhibition of the carbamoylcholine-stimulated increase was observed for pentobarbital, thiopental, phenobarbital, 5-(1,3-dimethylbutyl)-5-ethylbarbiturate, (+)- and (-)-5-ethyl-N-methyl-5-propylbarbituate and diphenylhydantoin. Similar concentrations of barbiturates and diphenylhydantoin were previously reported to inhibit the K+-stimulated Ca2+ influx, and therefore other agents that affect Ca2+ influx were tested to find whether they had any effect on 32P incorporation into these lipids. K+ (35 mM) increases 32P incorporation into phosphatidic acid, but to a smaller degree than 100 micrometer-carbamoylcholine, and its effect was inhibited by pentobarbital. Veratridine (75 micrometer) does not increase 32P incorporation into either phosphatidic acid or phosphatidylinositol, but did inhibit the carbamoylcholine-stimulated increase in 32P incorporation into phosphatidylinositol. The possible relationship between the phospholipid effect and stimulated Ca2+ influx is discussed. PMID:435289

  10. Phosphoglycolate phosphatase of spinach acts as a phosphoenzyme

    SciTech Connect

    Rose, Z.B.; Seal, S.N.

    1987-05-01

    When /sup 32/P-glycolate and phosphoglycolate phosphatase from spinach are mixed, /sup 32/P is incorporated into acid precipitated protein. Properties that relate this phosphorylation to the enzyme are: The K/sub m/ value for P-glycolate is similar for protein phosphorylation and substrate hydrolysis; the /sup 32/P appearing in the phosphoenzyme is diluted by unlabeled P-glycolate or the alternative substrate, ethyl-P; the activator Cl/sup -/ enhances the effectiveness of ethyl-P as a substrate and as an inhibitor of the formation of /sup 32/P-enzyme; and /sup 32/P is lost from the enzyme when /sup 32/P-glycolate is consumed. The acid denatured phosphorylated protein is a molecule of 34,000 Da, which is half of the molecular weight of the native protein and is similar in size to the labeled band that is seen on SDS-polyacrylamide gels. The enzyme-bound phosphoryl group appears to be an acyl-phosphate from its pH stability, being quite stable at pH 1, less stable at pH 5, and very unstable above pH 5. The bond is readily hydrolyzed in acid molybdate and it is sensitive to cleavage by hydroxylamine at pH 6.8. The demonstration of enzyme phosphorylation by /sup 32/P-glycolate resolves the dilemma presented by initial rate studies in which alternative substrates appeared to have different mechanisms.

  11. Novel methodology for the study of mercury methylation and reduction in sediments and water using 197Hg radiotracer.

    PubMed

    Ribeiro Guevara, Sergio; Zizek, Suzana; Repinc, Urska; Pérez Catán, Soledad; Jaćimović, Radojko; Horvat, Milena

    2007-03-01

    Mercury tracers are powerful tools that can be used to study mercury transformations in environmental systems, particularly mercury methylation, demethylation and reduction in sediments and water. However, mercury transformation studies using tracers can be subject to error, especially when used to assess methylation potential. The organic mercury extracted can be as low as 0.01% of the endogenous labeled mercury, and artefacts and contamination present during methylmercury (MeHg) extraction processes can cause interference. Solvent extraction methods based on the use of either KBr/H2SO4 or HCl were evaluated in freshwater sediments using 197Hg radiotracer. Values obtained for the 197Hg tracer in the organic phase were up to 25-fold higher when HCl was used, which is due to the coextraction of 197Hg2+ into the organic phase during MeHg extraction. Evaluations of the production of MeHg gave similar results with both MeHg extraction procedures, but due to the higher Hg2+ contamination of the controls, the uncertainty in the determination was higher when HCl was used. The Hg2+ contamination of controls in the HCl extraction method showed a nonlinear correlation with the humic acid content of sediment pore water. Therefore, use of the KBr/H2SO4 method is recommended, since it is free from these interferences. 197Hg radiotracer (T1/2=2.673 d) has a production rate that is about 50 times higher than that of 203Hg (T1/2=46.595 d), the most frequently used mercury radiotracer. Hence it is possible to obtain a similar level of performance to 203Hg when it is used it in short-term experiments and produced by the irradiation of 196Hg with thermal neutrons, using mercury targets with the natural isotopic composition. However, if the 0.15% natural abundance of the 196Hg isotope is increased, the specific activity of the 197Hg tracer can be significantly improved. In the present work, 197Hg tracer was produced from mercury 51.58% enriched in the 196Hg isotope, and a 340-fold

  12. Incorporation of P and C into Photosynthetic Products of Ankistrodesmus braunii as Affected by X-Rays.

    PubMed

    Jeschke, W D; Gimmler, H; Simonis, W

    1967-03-01

    The incorporation of (32)P and (14)C into organic compounds by Ankistrodesmus is strongly inhibited by X-rays. In the same phosphorylated compounds (32)P-incorporation apparently is more severely inhibited by X-rays than the (14)C-labelling. The (32)P-incorporation into organic compounds is more strongly inhibited than (32)P-labelling of inorganic phosphate in the cell. The inhibition of (32)P-incorporation into a number of compounds is strikingly uniform. It is concluded that the inhibition of (32)P-incorporation and of (14)C-incorporation into phosphorylated compounds in vivo is due to an uncoupling by X-rays of photophosphorylation as in vitro. The difference in X-ray sensitivity of (14)C- and (32)P-incorporation into one organic phosphorous compound is attributed to a dual action of X-rays on (32)P-incorporation in organic compounds (both via the uncoupling of photophosphorylation) and only a single effect on (14)C-incorporation and (32)P-labelling of inorganic phosphate. The effect of X-rays on (14)C-incorporation into organic compounds included inhibition in most cases but also stimulation as in the case of glycolic acid. These differences may be due to interference in the intercellular regulations following the application of X-rays. The inhibition of (14)C-incorporation in many cases exhibits different behaviour at low (<200 krad) and high doses. These changes are discussed on the assumption that at the lower doses X-rays cause uncoupling of photophosphorylation and at the higher doses an additional inhibition of electron transport.

  13. Malfunction of cardiac devices after radiotherapy without direct exposure to ionizing radiation: mechanisms and experimental data.

    PubMed

    Zecchin, Massimo; Morea, Gaetano; Severgnini, Mara; Sergi, Elisabetta; Baratto Roldan, Anna; Bianco, Elisabetta; Magnani, Silvia; De Luca, Antonio; Zorzin Fantasia, Anna; Salvatore, Luca; Milan, Vittorino; Giannini, Gianrossano; Sinagra, Gianfranco

    2016-02-01

    Malfunctions of cardiac implantable electronical devices (CIED) have been described after high-energy radiation therapy even in the absence of direct exposure to ionizing radiation, due to diffusion of neutrons (n) causing soft errors in inner circuits. The purpose of the study was to analyse the effect of scattered radiation on different types and models of CIED and the possible sources of malfunctions. Fifty-nine explanted CIED were placed on an anthropomorphous phantom of tissue-equivalent material, and a high-energy photon (15 MV) radiotherapy course (total dose = 70 Gy) for prostate treatment was performed. All devices were interrogated before and after radiation. Radiation dose, the electromagnetic field, and neutron fluence at the CIED site were measured. Thirty-four pacemakers (PM) and 25 implantable cardioverter-defibrillators (ICD) were analysed. No malfunctions were detected before radiation. After radiation a software malfunction was evident in 13 (52%) ICD and 6 (18%) PM; no significant electromagnetic field or photon radiations were detected in the thoracic region. Neutron capture was demonstrated by the presence of the (198)Au((197)Au + n) or (192)Ir((191)Ir + n) isotope activation; it was significantly greater in ICD than in PM and non-significantly greater in damaged devices. A greater effect in St Jude PM (2/2 damaged), Boston (9/11), and St Jude ICD (3/6) and in older ICD models was observed; the year of production was not relevant in PM. High-energy radiation can cause different malfunctions on CIED, particularly ICD, even without direct exposure to ionizing radiation due to scattered radiation of neutrons produced by the linear accelerator. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email: journals.permissions@oup.com.

  14. Enhancing the quality of radiographic images acquired with point-like gamma-ray sources through correction of the beam divergence and attenuation

    NASA Astrophysics Data System (ADS)

    Silvani, M. I.; Almeida, G. L.; Lopes, R. T.

    2014-11-01

    Radiographic images acquired with point-like gamma-ray sources exhibit a desirable low penumbra effects specially when positioned far away from the set object-detector. Such an arrangement frequently is not affordable due to the limited flux provided by a distant source. A closer source, however, has two main drawbacks, namely the degradation of the spatial resolution - as actual sources are only approximately punctual - and the non-homogeneity of the beam hitting the detector, which creates a false attenuation map of the object being inspected. This non-homogeneity is caused by the beam divergence itself and by the different thicknesses traversed the beam even if the object were an homogeneous flat plate. In this work, radiographic images of objects with different geometries, such as flat plates and pipes have undergone a correction of beam divergence and attenuation addressing the experimental verification of the capability and soundness of an algorithm formerly developed to generate and process synthetic images. The impact of other parameters, including source-detector gap, attenuation coefficient, ratio defective-to-main hull thickness and counting statistics have been assessed for specifically tailored test-objects aiming at the evaluation of the ability of the proposed method to deal with different boundary conditions. All experiments have been carried out with an X-ray sensitive Imaging Plate and reactor-produced 198Au and 165Dy sources. The results have been compared with other technique showing a better capability to correct the attenuation map of inspected objects unveiling their inner structure otherwise concealed by the poor contrast caused by the beam divergence and attenuation, in particular for those regions far apart from the vertical of the source.

  15. Relative toxicity of inhaled metal sulfate salts for pulmonary macrophages

    SciTech Connect

    Skornik, W.A.; Brain, J.D.

    1983-08-01

    The effects of metal sulfate aerosols on respiratory defense mechanisms in hamsters were studied. Pulmonary macrophage phagocytic rates were measured by determining the in vivo uptake of radioactive colloidal gold (/sup 198/Au) 1, 24, or 48 h after a single 4-h exposure. The concentrations of sulfate aerosols causing a 50% inhibition in pulmonary macrophage endocytosis (EC/sub 50/) were determined. When hamsters were exposed for 4 h to cupric sulfate (greater than or equal to 4.8 mg/m/sup 3/), zinc sulfate (greater than or equal to 3.1 mg/m/sup 3/), ferric sulfate (greater than or equal to 7.8 mg/m/sup 3/), or zinc ammonium sulfate (greater than or equal to 10.0 mg/m/sup 3/), macrophage endocytosis was significantly reduced 1 h after exposure compared with that in unexposed control animals. Although the response was variable, 24 h after exposures to the higher sulfate concentrations the percent of gold ingested by pulmonary macrophages remained depressed. By 48 h, the rate of macrophage endocytosis in hamsters had returned to normal control values except in hamsters exposed to 4.8 mg/m/sup 3/ cupric sulfate or 9.8 mg/m/sup 3/ ferric sulfate. These hamsters showed significant increases in phagocytosis. The EC/sub 50/ values in milligrams of sulfate per cubic meter for cupric sulfate, zinc sulfate, ferric sulfate, and zinc ammonium sulfate were 2.7, 4.5, 7.5, and 17.9, respectively. These results are negatively correlated with the ranking of sulfates using the criteria of relative irritant potency, as measured by increases in pulmonary flow resistance. Thus, rankings of related chemical structures are not absolute. Their relative toxicities vary depending on the end point selected.

  16. Relative toxicity of inhaled metal sulfate salts for pulmonary macrophages.

    PubMed

    Skornik, W A; Brain, J D

    1983-08-01

    The effects of metal sulfate aerosols on respiratory defense mechanisms in hamsters were studied. Pulmonary macrophage phagocytic rates were measured by determining the in vivo uptake of radioactive colloidal gold (198Au) 1, 24, or 48 h after a single 4-h exposure. The concentrations of sulfate aerosols causing a 50% inhibition in pulmonary macrophage endocytosis (EC50) were determined. When hamsters were exposed for 4 h to cupric sulfate (greater than or equal to 4.8 mg/m3), zinc sulfate (greater than or equal to 3.1 mg/m3), ferric sulfate (greater than or equal to 7.8 mg/m3), or zinc ammonium sulfate (greater than or equal to 10.0 mg/m3), macrophage endocytosis was significantly reduced 1 h after exposure compared with that in unexposed control animals. Although the response was variable, 24 h after exposures to the higher sulfate concentrations the percent of gold ingested by pulmonary macrophages remained depressed. By 48 h, the rate of macrophage endocytosis in hamsters had returned to normal control values except in hamsters exposed to 4.8 mg/m3 cupric sulfate or 9.8 mg/m3 ferric sulfate. These hamsters showed significant increases in phagocytosis. The EC50 values in milligrams of sulfate per cubic meter for cupric sulfate, zinc sulfate, ferric sulfate, and zinc ammonium sulfate were 2.7, 4.5, 7.5, and 17.9, respectively. These results are negatively correlated with the ranking of sulfates using the criteria of relative irritant potency, as measured by increases in pulmonary flow resistance. Thus, rankings of related chemical structures are not absolute. Their relative toxicities vary depending on the end point selected.

  17. Experimental determination of the regional deposition of aerosol particles in the human respiratory tract

    SciTech Connect

    Stahlhofen, W.; Gebhart, J.; Heyder, J.

    1980-06-01

    The experimental techniques and the results of inhalation studies with radioaerosols on normal non-smokers for mouth-breathing are described and discussed. Monodisperse iron oxide particles tagged with /sup 198/Au are produced with a spinning top generator in the aerodynamic size range between 1 to 10 ..mu..m. An aerosol inhalation apparatus enables the subjects to breathe under standardized conditions with respect to tidal volume and breathing frequency. The calculation of total deposition is based upon measurements of the number of in- and exhaled particles per breath by means of photometric methods and pneumotachography. The retention of the radioactive particles present in the body after aerosol administration is measured with a body counter designed and constructed for these experiments. Retention measurements as functions of time after inhalation are carried out in extrathoracic-, chest- and stomach-position. The body counter consists of four shielded NaF(Tl)-dectors. Characteristic feature of the body counter is its low sensitivity to neighboring organs and to neighboring regions within the respiratory tract. For the evaluation of extrathoracic deposition, the activity measured in the stomach immediately after inhalation is added to extrathoracic activity. The elimination of material from the chest is found to be much slower for the material deposited in the alveolar region than for the amount deposited in the tracheobronchial tree. This allows the intrathoracic deposition to be divided into tracheolbronchial and alveolar deposition by means of the different slopes of the normalized chest retention function. Different normalized chest retention functions are presented and analyzed with respect to their different elimination rats belonging to the tracheobronchial and alveolar region. Total, tracheobronchial, alveolar and extrathoracic deposition data are reported in the aerodynamic diameter range between 1 and 10 ..mu..m.

  18. Thermal neutron capture cross-section and resonance integral measurements of 139La(n, γ)140La and 140Ce(n, γ)141Ce using a Am-Be neutron source

    NASA Astrophysics Data System (ADS)

    Panikkath, Priyada; Mohanakrishnan, P.

    2017-03-01

    Thermal neutron capture cross-sections and resonance integrals of 139La(n, γ)140La and 140Ce (n, γ)141Ce are measured with respect to reference reactions 197Au(n, γ)198Au and 55Mn(n, γ)56Mn using the neutron activation technique. Measurements are carried out using neutrons from an Am-Be source located inside a concrete bunker. Two different methods are used for determining self-shielding factors of activation foils as well as for finding the epithermal neutron spectrum shape factor. For 139 La with reference to 197 Au and 55 Mn the measured thermal cross sections are 9.24 ± 0.25 b and 9.28 ± 0.37 b, respectively, while the measured resonance integrals are 12.18 ± 0.67 b and 11.81 ± 0.94 b, respectively. For 140 Ce with reference to 197 Au and 55 Mn the measured thermal cross sections are 0.44 ± 0.01 b and 0.44 ± 0.02 b, respectively, while the measured resonance integrals are 0.55 ± 0.03 b and 0.54 ± 0.04 b, respectively. The present measurements are compared with earlier measurements and evaluations. Presently estimated values confirm the established 139La(n, γ)140La cross-sections. The presently measured thermal capture cross-section 140Ce(n, γ)141Ce , though lower than the evaluated data, is having higher accuracy compared to previous measurements with large uncertainties. The resonance integral measured is higher (like most previous measurements) than most evaluations requiring a revision of the evaluated data.

  19. Activities on Nuclear Data Measurements at Pohang Neutron Facility

    NASA Astrophysics Data System (ADS)

    Kim, Guinyun

    2009-03-01

    We report the activities of the Pohang Neutron Facility which consists of an electron linear accelerator, a water-cooled Ta target, and a 12-m time-of-flight path. It has been equipped with a four-position sample changer controlled remotely by a CAMAC data acquisition system, which allows simultaneous accumulation of the neutron time of flight spectra from 4 different detectors. It can be possible to measure the neutron total cross-sections in the neutron energy range from 0.1 eV to few hundreds eV by using the neutron time-of-flight method. A 6LiZnS(Ag) glass scintillator was used as a neutron detector. The neutron flight path from the water-cooled Ta target to the neutron detector was 12.1 m. The background level was determined by using notch-filters of Co, In, Ta, and Cd sheets. In order to reduce the gamma rays from bremsstrahlung and those from neutron capture, we employed a neutron-gamma separation system based on their different pulse shapes. The present measurements of several samples (Ta, Mo) are in general agreement with the evaluated data in ENDF/B-VI. We measured the thermal neutron capture cross-sections and the resonance integrals of the 186W(n,γ)187W reaction and the 98Mo(n,γ)99Mo reaction by the activation method using the 197Au(n,γ)198Au monitor reaction as a single comparator. We also report the isomeric yield ratios for the 44 m, gSc isomeric pairs produced from four different photonuclear reactions 45Sc(γ,n)44m,gSc, natTi(γ,xn1p)44m,gSc, natFe(γ,xn5p)52m,gMn, and 103Rh(γ,4n)99m,gRh by using the activation method.

  20. Effect of sulfur dioxide on pulmonary macrophage endocytosis at rest and during exercise

    SciTech Connect

    Skornik, W.A.; Brain, J.D. )

    1990-09-01

    Inhaled SO2 may cause damage by injuring upper airways. To what extent can SO2 also alter pulmonary macrophage function in the parenchyma and what is the impact of exercise We studied the effect of SO2 on pulmonary macrophage endocytosis in resting and in exercising animals by measuring the rates of macrophage endocytosis in situ for 1 h of a test particle of insoluble radioactive colloidal gold (198Au), 1, 24, or 48 h after inhalation exposure to SO2. Resting hamsters exposed for 4 h to 50 ppm SO2 had no significant reduction in macrophage endocytosis compared with air-breathing control hamsters. However, if hamsters were exposed to the same concentration of SO2 while continuously running (40 min at 0.9 km/h), macrophage endocytosis was significantly reduced 1 h after exposure even though the exposure time was only one-sixth as long. Twenty-four hours later, the percentage of gold ingested by pulmonary macrophages remained significantly depressed. By 48 h, the rate had returned to control values. Exercise alone did not affect endocytosis. Hamsters exposed to 50 ppm SO2, with or without exercise, also showed significant reductions in the number of lavaged macrophages. This decrease was greatest and most persistent in the SO2 plus exercise group. These data indicate that even when animals are exposed to water-soluble gases, which are normally removed by the upper airways, exercise can potentiate damage to more peripheral components of the pulmonary defense system such as the macrophage.

  1. Cardiac and non-cardiac malformations produced by Mercury in hamsters. [None

    SciTech Connect

    Gale, T.F.

    1980-11-01

    The susceptibility of the developing mammalian embryo to the adverse effects of mercury is well documented. A variety of organic mercury compounds have been demonstrated to produce embryotoxic effects in experimental animals. HARADA recently summarized the reports of human intrauterine methylmercury poisoning, i.e., congenital Minamata disease, resulting from the ingestion of contaminated food. Ongoing studies in this laboratory have involved several different aspects of the embryotoxicity produced by inorganic mercury in hamsters including a dose response study, the interaction of mercuric acetate with cadmium and zinc, the effect of different routes of administration, the placental permeability of /sup 203/Hg and the embryotoxic response in several different hamster strains. Little is known regarding a human syndrome of congenital malformations characterized by ectopia cordis, internal cardiac defects and abnormalities of the diaphragm and ventral body wall. Most papers regarding this human syndrome are clinical reports describing the characteristics and management of specific cases; only speculative information is provided regarding etiology and possible embryopathic mechanisms. The observation that a similar syndrome, which will be designated CNC for cardiac and non-cardiac malformations, can be produced by mercury in hamsters prompted the present study. The specific goals of this study were 1) to study the effect of treating pregnant hamsters at different times during embryonic organogenesis to determine the time which produces the highest incidence of the CNC syndrome and whether different treatment times modify the morphological characteristics of the inclusive malformations and 2) to study the structural features of all mercury-induced external and internal abnormalities of the CNC syndrome in late gestation fetuses.

  2. The accumulation of lead and mercury from seawater and their depuration by eggs of the spotted dogfish Scyliorhinus canicula (Chondrichthys).

    PubMed

    Jeffree, R A; Oberhansli, F; Teyssie, J-L

    2008-10-01

    Radiotracer experiments using (210)Pb and (203)Hg demonstrated that eggs of the spotted dogfish Scyliorhinus canicula absorbed lead and inorganic mercury directly from seawater over 21 days of experimental exposure, attaining total egg concentration factors (CFs) relative to water of approximately 400 for Pb and 180 for Hg, predominantly (> or =98%) due to their accumulation by the collagenous egg case. The rates of accumulation of both Pb and Hg by the total egg were significantly (P < 0.0001) reduced by its increasing age since parturition, whereas only the rate of depuration of Pb was reduced (P < 0.0001) with increasing age; these effects indicate a declining chemical reactivity of the egg case that may be due to the continued tanning of the case following parturition. The egg case per se, attained average CFs of about 1,500 and 850 for Pb and Hg, respectively. Both Pb and Hg showed declining concentration gradients from the exterior to the interior membranes of the wall of the egg case; CFs for Pb declined from 3,500 to 2,000 and for Hg from 5,000 to 500. Comparison of concentrations in separate membranes also demonstrated significant (P < or = 0.01) depurations of Hg from the external and internal membranes during the loss experiments. The presence of radiotracers of Pb and Hg in the internal components of the egg at the end of uptake phase, and prior to the opening of the apertures, confirmed the permeability of the egg case wall to them, consistent with their observed gradients in it. The average CFs for all embryos at the end of the uptake experiment were 34 and 44 for Pb and Hg, respectively, but were significantly (P < 0.001) enhanced for Hg by a factor of 6 in the older eggs. The accumulatory and kinetic characteristics of the egg-case may operate to optimize the exposure of embryos to Pb and Hg following episodic contaminant events in coastal habitats.

  3. Methylmercury Bioaccumulation, Transformation, and Trophic Transfer in Marine Plankton Assemblages

    NASA Astrophysics Data System (ADS)

    Lee, C. S.; Fisher, N. S.

    2016-02-01

    Few studies have quantified the bioconcentration of methylmercury (MeHg) in marine phytoplankton from seawater, even though this is by far the largest bioaccumulation step in aquatic organisms. Aquatic animals acquire MeHg mainly from dietary exposure and it is important to evaluate the bioaccumulation of this compound in planktonic organisms that form the base of marine food webs. We used a gamma-emitting radioisotope, 203Hg, to assess the rate and extent of MeHg uptake in marine diatoms, dinoflagellates, coccolithophores, cryptophytes chlorophytes, and cyanobacteria held in unialgal cultures under varying temperature, light and nutrient conditions. For experimental conditions in which cells were exposed to MeHg at 300 pM, the uptake rates in all species ranged from 0.001 to 0.034 atto-mol MeHg µm-2 cell surface h-1 and reached steady state within 2 d. Volume concentration factors (VCFs) ranged from 0.3 to 40 x 105 for the different species. Temperature, light and nutrient conditions had no direct effect on cellular MeHg uptake but ultimately affected growth of the cells, resulting in greater suspended particulate matter and associated MeHg. VCFs strongly correlated with cell surface area to volume ratios in all species. Nearly 40 % of the MeHg was released into the air from coccolithophore cultures within 4 d, but <10 % from other algal cultures. Assimilation efficiencies of MeHg from different phytoplankton diets in a marine copepod (Acartia tonsa) ranged from 74 to 92%, directly proportional to the cytoplasmic partitioning of MeHg in the phytoplankton cells. MeHg uptake in copepods from the aqueous phase was low and modeling shows that nearly all the MeHg acquired by this zooplankter is from diet. Herbivorous zooplankton can be an important link from phytoplankton at the base of the food web to fish higher in the food chain.

  4. Methyl mercury uptake by diverse marine phytoplankton and trophic transfer to zooplankton

    NASA Astrophysics Data System (ADS)

    Lee, C. S.; Fisher, N. S.

    2014-12-01

    While it is well known that methylmercury (MeHg) biomagnifies in aquatic food chains, few studies have quantified its bioaccumulation in marine phytoplankton from seawater, even though that is overwhelmingly the largest bioaccumulation step. Aquatic animals acquire MeHg mainly from dietary exposure and it is important to evaluate the bioaccumulation of this compound in planktonic organisms that form the base of marine food webs. We used a gamma-emitting radioisotope, 203Hg, to assess the rate and extent of MeHg uptake in marine diatoms, dinoflagellates, coccolithophores, cryptophytes chlorophytes, and cyanobacteria held in unialgal cultures under varying temperature and light conditions. For experimental conditions in which the dissolved MeHg was at 300 pM, the uptake rates in all species ranged from 0.004 to 0.75 amol Hg μm-3 cell volume d-1 and reached steady state within 2 d. Volume concentration factors (VCFs) ranged from 0.4 to 60 x 105 for the different species. Temperature and light conditions had no direct effect on cellular MeHg uptake but ultimately affected growth of the cells, resulting in greater suspended particulate matter and associated MeHg. VCFs strongly correlated with cell surface area to volume ratios in all species. Assimilation efficiencies of MeHg from phytoplankton food (Thalassiosira pseudonana, Dunaliella tertiolecta and Rhodomonas salina) in a marine copepod grazer (Acartia tonsa) ranged from 74 to 92%, directly proportional to the cytoplasmic partitioning of MeHg in the phytoplankton cells. MeHg uptake in copepods from the aqueous phase was low and modeling shows that nearly all the MeHg acquired by this zooplankter is from diet. Herbivorous zooplankton can be an important link from phytoplankton at the base of the food web to fish higher in the food chain.

  5. Constants for mercury binding by organic matter isolates from the Florida Everglades

    USGS Publications Warehouse

    Benoit, J.M.; Mason, R.P.; Gilmour, C.C.; Aiken, G.R.

    2001-01-01

    Dissolved organic matter (DOM) has been implicated as an important complexing agent for Hg that can affect its mobility and bioavailability in aquatic ecosystems. However, binding constants for natural Hg-DOM complexes are not well known. We employed a competitive ligand approach to estimate conditional stability constants for Hg complexes with DOM isolates collected from Florida Everglades surface waters. The isolates examined were the hydrophobic fraction of DOM from a eutrophic, sulfidic site (F1-HPoA) and the hydrophilic fraction from an oligotrophic, low-sulfide site (2BS-HPiA). Our experimental determinations utilized overall octanol-water partitioning coefficients (Dow) for 203Hg at 0.01 M chloride and across pH and DOM concentration gradients. Use of this radioisotope allowed rapid determinations of Hg concentrations in both water and octanol phases without problems of matrix interference. Conditional stability constants (1 = 0.06, 23??C) were log K??? = 11.8 for F1-HPoA and log K' = 10.6 for 2BS-HPiA. These are similar to previously published stability constants for Hg binding to low-molecular-weight thiols. Further, F1-HPoA showed a pH-dependent decline in Dow that was consistent with models of Hg complexation with thiol groups as the dominant Hg binding sites in DOM. These experiments demonstrate that the DOM isolates are stronger ligands for Hg than chloride ion or ethylenediamine-tetraacetic acid. Speciation calculations indicate that at the DOM concentrations frequently measured in Everglades, 20 to 40 ??M, significant complexation of Hg by DOM would be expected in aerobic (sulfide-free) surface waters. Copyright ?? 2001 Elsevier Science Ltd.

  6. Radiotracer Dilution Method for Mercury Inventory Study in Electrolytic Cells

    SciTech Connect

    Sugiharto; Su'ud, Zaki; Kurniadi, Rizal; Waris, Abdul; Santoso, Sigit Budi; Abidin, Zainal; Santoso, Gatot Budi

    2010-06-22

    Purpose of the experiment is to demonstrate feasibility the use of radiotracer to measure weight of mercury in electrolytic cells of soda industry. The weight of mercury in each cell of the plant is designed approximately 1700 kg. Radiotracer is prepared by mixing {sup 203}Hg radioactive mercury with 2400 g of inactive mercury in a bath. The respective precisely weighted mercury aliquots to be injected into the cells are prepared by pouring approximately 130 g of radioactive mercury taken from the bath into 13 standard vials, in accordance with the number of the cells tested. Four standard references prepared by further dilution of {+-}2 g active mercury taken from the bath to obtain the dilution factors range of 12,000 to 20,000 from which the calibration graph is constructed. The injection process is conducting by pouring the radioactive mercury from aliquots into the flowing mercury at the inlet side of the cell and allows them to mix thoroughly. It is assumed that the mass of the radiotracer injected into a closed system remains constant, at least during the period of the test. From this experiment it was observed that the mixing time is two days after injection of radioactive mercury. The inactive mercury in each electrolytic cell calculated by the radiotracer method is of the range 1351.529 kg to 1966.354 kg with maximum error (95% confidence) is 1.52 %. The accuracy of measurement of the present method is better than gravimetric one which accounts 4 % of error on average.

  7. Mercury toxicokinetics-dependency on strain and gender

    SciTech Connect

    Ekstrand, Jimmy; Nielsen, Jesper B.; Havarinasab, Said; Zalups, Rudolfs K.; Soederkvist, Peter; Hultman, Per

    2010-03-15

    Mercury (Hg) exposure from dental amalgam fillings and thimerosal in vaccines is not a major health hazard, but adverse health effects cannot be ruled out in a small and more susceptible part of the exposed population. Individual differences in toxicokinetics may explain susceptibility to mercury. Inbred, H-2-congenic A.SW and B10.S mice and their F1- and F2-hybrids were given HgCl{sub 2} with 2.0 mg Hg/L drinking water and traces of {sup 203}Hg. Whole-body retention (WBR) was monitored until steady state after 5 weeks, when the organ Hg content was assessed. Despite similar Hg intake, A.SW males attained a 20-30% significantly higher WBR and 2- to 5-fold higher total renal Hg retention/concentration than A.SW females and B10.S mice. A selective renal Hg accumulation but of lower magnitude was seen also in B10.S males compared with females. Differences in WBR and organ Hg accumulation are therefore regulated by non-H-2 genes and gender. Lymph nodes lacked the strain- and gender-dependent Hg accumulation profile of kidney, liver and spleen. After 15 days without Hg A.SW mice showed a 4-fold higher WBR and liver Hg concentration, but 11-fold higher renal Hg concentration, showing the key role for the kidneys in explaining the slower Hg elimination in A.SW mice. The trait causing higher mercury accumulation was not dominantly inherited in the F1 hybrids. F2 mice showed a large inter-individual variation in Hg accumulation, showing that multiple genetic factors influence the Hg toxicokinetics in the mouse. The genetically heterogeneous human population may therefore show a large variation in mercury toxicokinetics.

  8. Experimental response function of a 3 in×3 in NaI(Tl) detector by inverse matrix method and effective atomic number of composite materials by gamma backscattering technique.

    PubMed

    Kiran, K U; Ravindraswami, K; Eshwarappa, K M; Somashekarappa, H M

    2016-05-01

    Response function of a widely used 3in×3in NaI(Tl) detector is constructed to correct the observed pulse height distribution. A 10×10 inverse matrix is constructed using 7 mono-energetic gamma sources ((57)Co, (203)Hg, (133)Ba, (22)Na, (137)Cs, (54)Mn and (65)Zn) which are evenly spaced in energy scale to unscramble the observed pulse height distribution. Bin widths (E)(1/2) of 0.01 (MeV)(1/2) are used to construct the matrix. Backscattered photons for an angle of 110° are obtained from a well-collimated 0.2146GBq (5.8mCi) (137)Cs gamma source for carbon, aluminium, iron, copper, granite and Portland cement. For each observed spectrum, single scattered spectrum is constructed analytically using detector parameters like FWHM, photo-peak efficiency and peak counts. Response corrected multiple scattered photons are extracted from the observed pulse height distribution by dividing the spectrum into a 10 ×1 matrix. Saturation thicknesses of carbon, aluminium, iron, copper, granite and Portland cement are found out. Variation of multiple scattered photons as a function of target thickness are simulated using MCNP code. A relationship between experimental and simulated saturation thicknesses of carbon, aluminium, iron and copper is obtained as a function of atomic number. Using this relation, effective atomic numbers of granite and Portland cement are obtained from interpolation method. Effective atomic numbers of granite and Portland cement are also obtained by theoretical equation using their elemental composition and comparing with the experimental and simulated results.

  9. Multidrug Resistance Proteins and the Renal Elimination of Inorganic Mercury Mediated by 2,3-Dimercaptopropane-1-Sulfonic Acid and Meso-2,3-dimercaptosuccinic Acid

    PubMed Central

    Bridges, Christy C.; Joshee, Lucy; Zalups, Rudolfs K.

    2008-01-01

    Current therapies for inorganic mercury (Hg2+) intoxication include administration of a metal chelator, either 2,3-dimercaptopropane-1-sulfonic acid (DMPS) or meso-2,3-dimercaptosuccinic acid (DMSA). After exposure to either chelator, Hg2+ is rapidly eliminated from the kidneys and excreted in the urine, presumably as an S-conjugate of DMPS or DMSA. The multidrug resistance protein 2 (Mrp2) has been implicated in this process. We hypothesize that Mrp2 mediates the secretion of DMPS- or DMSA-S-conjugates of Hg2+ from proximal tubular cells. To test this hypothesis, the disposition of Hg2+ was examined in control and Mrp2-deficient TR− rats. Rats were injected i.v. with 0.5 μmol/kg HgCl2 containing 203Hg2+. Twenty-four and 28 h later, rats were injected with saline, DMPS, or DMSA. Tissues were harvested 48 h after HgCl2 exposure. The renal and hepatic burden of Hg2+ in the saline-injected TR− rats was greater than that of controls. In contrast, the amount of Hg2+ excreted in urine and feces of TR− rats was less than that of controls. DMPS, but not DMSA, significantly reduced the renal and hepatic content of Hg2+ in both groups of rats, with the greatest reduction in controls. A significant increase in urinary and fecal excretion of Hg2+, which was greater in the controls, was also observed following DMPS treatment. Experiments utilizing inside-out membrane vesicles expressing MRP2 support these observations by demonstrating that DMPS- and DMSA-S-conjugates of Hg2+ are transportable substrates of MRP2. Collectively, these data support a role for Mrp2 in the DMPS- and DMSA-mediated elimination of Hg2+ from the kidney. PMID:17940195

  10. Mercury cycling in stream ecosystems. 2. Benthic methylmercury production and bed sediment - Pore water partitioning

    USGS Publications Warehouse

    Marvin-DiPasquale, M.; Lutz, M.A.; Brigham, M.E.; Krabbenhoft, D.P.; Aiken, G.R.; Orem, W.H.; Hall, B.D.

    2009-01-01

    Mercury speciation, controls on methylmercury (MeHg) production, and bed sediment - pore water partitioning of total Hg (THg) and MeHg were examined in bed sediment from eight geochemically diverse streams where atmospheric deposition was the predominant Hg input. Across all streams, sediment THg concentrations were best described as a combined function of sediment percent fines (%fines; particles < 63 ??m) and organic content. MeHg concentrations were best described as a combined function of organic content and the activity of the Hg(II)-methylating microbial community and were comparable to MeHg concentrations in streams with Hg inputs from industrial and mining sources. Whole sediment tin-reducible inorganic reactive Hg (Hg(II)R) was used as a proxy measure for the Hg(II) pool available for microbial methylation. In conjunction with radiotracer-derived rate constants of 203Hg(II) methylation, Hg(II)R was used to calculate MeHg production potential rates and to explain the spatial variability in MeHg concentration. The %Hg(II)R (of THg) was low (2.1 ?? 5.7%) and was inversely related to both microbial sulfate reduction rates and sediment total reduced sulfur concentration. While sediment THg concentrations were higher in urban streams, %MeHg and %Hg(II)R were higher in nonurban streams. Sediment pore water distribution coefficients (log Kd's) for both THg and MeHg were inversely related to the log-transformed ratio of pore water dissolved organic carbon (DOC) to bed sediment %fines. The stream with the highest drainage basin wetland density also had the highest pore water DOC ?? 2009 American Chemical Society.

  11. Sexual differences in the excretion of organic and inorganic mercury by methyl mercury-treated rats

    SciTech Connect

    Thomas, D.J.; Fisher, H.L.; Sumler, M.R.; Mushak, P.; Hall, L.L.

    1987-06-01

    Adult male and female Long Evans rats received 1 mumole of methyl (/sup 203/Hg) mercuric chloride per kilogram sc. Whole-body retention of mercury and excretion of organic and inorganic mercury in urine and feces were monitored for 98 days after dosing. Females cleared mercury from the body more rapidly than did males. The major route of mercury excretion was feces. By 98 days after dosing, cumulative mercury excretion in feces accounted for about 51% of the dose in males and about 54% of the dose in females. For both sexes, about 33% of the dose was excreted in feces as inorganic mercury. Cumulative excretion of organic mercury in feces accounted for about 18 and 21% of the dose in males and females, respectively. Urinary excretion of mercury was quantitatively a smaller route for mercury clearance but important sexual differences in loss by this route were found. Over the 98-day experimental period, males excreted in urine about 3.2% of the dose and females excreted 7.5%. Cumulative organic Hg excretion in urine accounted for 1.8% of the dose in males and 5.3% of the dose in females. These sexual differences in urinary and fecal excretion of organic and inorganic mercury following methyl mercury treatment were consistent with previous reports of sexual differences in mercury distribution and retention in methyl mercury-treated rats, particularly sexual differences in organic mercury uptake and retention in the kidney. Relationships between body burdens of organic or inorganic Hg and output of these forms of Hg in urine and feces were also found to be influenced by the interval after MeHg treatment and by sex. Relationship between concentration of Hg in liver and feces and in kidney and urine differed for organic and inorganic Hg and depended upon sexual status and interval after MeHg treatment.

  12. Occurrence of methylated mercury in a terrestrial food chain

    SciTech Connect

    Huckabee, J.W.; Lucas, D.M.; Baird, J.M.

    1981-10-01

    The presence and transfer of methylated mercury (Me-Hg) in a terrestrial food chain were determined by radiotracer laboratory experiments and chemical analyses of field-collected organisms. (/sup 203/Hg)MeHg-tagged fescue grass (Festuca sp.) at 0.65 ..mu..g MeHg/g grass (wet wt) was fed to cotton rats (Sigmodon hisidus). The rats had an assimillation efficiency (A/sub e/) for MeHg of 99% and a biological half-life (t/sub b/) of 9.5 days. Control S. hispidus and Festuca collected from the same populations as the experimental organisms were analyzed for MeHg and found to have, for the S. hispidus, 1.4 +/- 0.2 ng/g, 1 SE, and 1.3 +/- 0.3 ng/g, 1 SE, in skeletal muscle and liver, respectively; and for Festuca (composite sample) 0.6 ng/g. Total mercury (..sigma..Hg) for the same tissues was 119.4 +/- 39 ng/g, 1 SE, 34.6 +/- 8.9 ng/g, 1 SE; and 71 ng/g (composite sample), respectively. Calculations with the experimental and analytical MeHg values indicated that the feral S. hispidus would, at equilibrium, have a whole-body MeHg concentration of 1.89 ng/g. These results, compared with literature reports, indicate that feral rodents have concentration ranges of approx.50 to approx.160 ng/g for ..sigma..Hg, and approx.1 to approx.7 ng/g for MeHg. Environmental levels of mercury are likely to increase significantly; further environmental measurements and especially appropriate toxicity tests are waranted to assess the importance of MeHg in terrestrial ecosystems.

  13. NUCLEAR MEDICINE PRACTICES IN THE 1950s THROUGH THE mid-1970s AND OCCUPATIONAL RADIATION DOSES TO TECHNOLOGISTS FROM DIAGNOSTIC RADIOISOTOPE PROCEDURES

    PubMed Central

    Drozdovitch, Vladimir; Brill, Aaron B.; Mettler, Fred A.; Beckner, William M.; Goldsmith, Stanley J.; Gross, Milton D.; Hays, Marguerite T.; Kirchner, Peter T.; Langan, James K.; Reba, Richard C.; Smith, Gary T.; Bouville, André; Linet, Martha S.; Melo, Dunstana R.; Lee, Choonsik; Simon, Steven L.

    2014-01-01

    Data on occupational radiation exposure from nuclear medicine procedures for the time period of the 1950s through the 1970s is important for retrospective health risk studies of medical personnel who conducted those activities. However, limited information is available on occupational exposure received by physicians and technologists who performed nuclear medicine procedures during those years. To better understand and characterize historical radiation exposures to technologists, we collected information on nuclear medicine practices in the 1950s, 1960s, and 1970s. To collect historical data needed to reconstruct doses to technologists, a focus group interview was held with experts who began using radioisotopes in medicine in the 1950s and the 1960s. Typical protocols and descriptions of clinical practices of diagnostic radioisotope procedures were defined by the focus group and were used to estimate occupational doses received by personnel, per nuclear medicine procedure, conducted in the 1950s-1960s using radiopharmaceuticals available at that time. The radionuclide activities in the organs of the reference patient were calculated using the biokinetic models described in ICRP Publication 53. Air kerma rates as a function of distance from a reference patient were calculated by Monte Carlo radiation transport calculations using a hybrid computational phantom. Estimates of occupational doses to nuclear medicine technologists per procedure were found to vary from less than 0.01 μSv (thyroid scan with 1.85 MBq of administered 131I-iodide) to 0.4 μSv (brain scan with 26 MBq of 203Hg-chlormerodin). Occupational doses for the same diagnostic procedures starting in the mid-1960s but using 99mTc were also estimated. The doses estimated in this study show that the introduction of 99mTc resulted in an increase in occupational doses per procedure. PMID:25162420

  14. Uptake of mercury by the hair of methylmercury-treated newborn mice

    SciTech Connect

    Shi, Chenyang; Lane, A.T.; Clarkson, T.W. )

    1990-04-01

    Human hair has unique advantages in monitoring environmental exposures to methyl-mercury. Using newborn Balb/c mice as a model system, the incorporation of methylmercury into the hair was studied and compared with methylmercury distributions in other tissues. Newborn mice were given intraperitoneal injections of {sup 203}Hg-labeled methylmercury at designated times according to hair growth stages of the mouse. Animals were sacrificed 2 days after dosing. Distribution of mercury in pelt and other tissues was measured. The level of mercury in pelt was found to correlate with hair growth. The amount of mercury in pelt peaked when hair growth was most rapid and the total amount of mercury in pelt was significantly higher than that in other tissues, constituting 40% of the whole body burden. However, when the hair ceased growing, the amount of mercury in pelt dramatically dropped to 4% of whole body burden and mercury concentrations in other tissues except brain were elevated. Autoradiographic studies with tritium-labeled methylmercury demonstrated that methylmercury concentrated in hair follicles in the skin. Within hair follicles and hairs, methylmercury accumulated in regions that are rich in high-sulfur proteins. The uptake of inorganic mercury (administered as HgCl{sub 2}) by pelt was also compared with that of methylmercury. The amount of inorganic mercury found in pelt was less than one-half that of methylmercury in animals with growing hair. Cessation of hair growth did not decrease the inorganic mercury level in pelt to the same extent as in the case of methylmercury.

  15. MRP2 and the DMPS- and DMSA-mediated elimination of mercury in TR(-) and control rats exposed to thiol S-conjugates of inorganic mercury.

    PubMed

    Bridges, Christy C; Joshee, Lucy; Zalups, Rudolfs K

    2008-09-01

    Cysteine (Cys) and homocysteine (Hcy)-S-conjugates of inorganic mercury (Hg2+) are transportable species of Hg2+ that are taken up readily by proximal tubular cells. The metal chelators, 2,3-dimercaptopropane-1-sulfonic acid (DMPS) and meso-2,3-dimercaptosuccinic acid (DMSA), have been used successfully to extract Hg2+ from these cells, presumably via the multidrug resistance protein (Mrp2). In the current study, we tested the hypothesis that Mrp2 is involved in the DMPS- and DMSA-mediated extraction of Hg2+ following administration of Hg2+ as an S-conjugate of Cys or Hcy. To test this hypothesis, control and TR(-) (Mrp2-deficient) rats were injected with 0.5 micromol/kg HgCl2 (containing 203Hg2+) conjugated to 1.25 micromol/kg Cys or Hcy. After 24 and 28 h, rats were treated with saline or 100 mg/kg DMPS or DMSA. Tissues were harvested 48 h after Hg2+ exposure. The renal and hepatic burden of Hg2+ was greater in saline-injected TR- rats than in corresponding controls. Accordingly, the content of Hg2+ in the urine and feces was less in TR- rats than in controls. Following treatment with DMPS or DMSA, the renal content of Hg2+ in both groups of rats was reduced significantly and the urinary excretion of Hg2+ was increased. In liver, the effect of each chelator appeared to be dependent upon the form in which Hg2+ was administered. In vitro experiments provide direct evidence indicating that DMPS and DMSA-S-conjugates of Hg2+ are substrates for Mrp2. Overall, these data support our hypothesis that Mrp2 is involved in the DMPS and DMSA-mediated extraction of the body burden of Hg2+.

  16. Leach tests on grouts made with actual and trace metal-spiked synthetic phosphate/sulfate waste

    SciTech Connect

    Serne, R.J.; Martin, W.J.; LeGore, V.L.; Lindenmeier, C.W.; McLaurine, S.B.; Martin, P.F.C.; Lokken, R.O.

    1989-10-01

    Pacific Northwest Laboratory conducted experiments to produce empirical leach rate data for phosphate-sulfate waste (PSW) grout. Effective diffusivities were measured for various radionuclides ({sup 90}Sr, {sup 99}Tc, {sup 14}C, {sup 129}I, {sup 137}Cs, {sup 60}Co, {sup 54}Mn, and U), stable major components (NO{sub 3}{sup {minus}}, SO{sub 4}{sup 2{minus}}, H{sub 3}BO{sub 3}, K and Na) and the trace constituents Ag, As, Cd, Hg, Pb, and Se. Two types of leach tests were used on samples of actual PSW grout and synthetic PSW grout: the American Nuclear Society (ANS) 16.1 intermittent replacement leach test and a static leach test. Grout produced from both synthetic and real PSW showed low leach rates for the trace metal constituents and most of the waste radionuclides. Many of the spiked trace metals and radionuclides were not detected in any leachates. None of the effluents contained measurable quantities of {sup 137}Cs, {sup 60}Co, {sup 54}Mn, {sup 109}Cd, {sup 51}Cr, {sup 210}Pb, {sup 203}Hg, or As. For those trace species with detectable leach rates, {sup 125}I appeared to have the greatest leach rate, followed by {sup 99}Tc, {sup 75}Se, and finally U, {sup 14}C, and {sup 110m}Ag. Leach rates for nitrate are between those for I and Tc, but there is much scatter in the nitrate data because of the very low nitrate inventory. 32 refs., 6 figs., 15 tabs.

  17. Amanita muscaria and Amanita pantherina poisoning: two syndromes.

    PubMed

    Vendramin, Andreja; Brvar, Miran

    2014-11-01

    Amanita muscaria contains more excitatory ibotenic acid and less depressant muscimol compared to Amanita pantherina. In this study A. muscaria poisoned patients were more often confused (26/32, p = 0.01) and agitated (20/32, p = 0.03) compared to those poisoned with A. pantherina (8/17 and 5/17). Patients poisoned with A. pantherina were more commonly comatose (5/17) compared to those poisoned with A. muscaria (2/32) (p = 0.03). In conclusion, the so-called ibotenic or pantherina-muscaria syndrome might be divided into two subtypes.

  18. Fibronectin from chicken embryo fibroblasts contains covalently bound phosphate

    PubMed Central

    1979-01-01

    Fibronectin isolated from cultures of chicken embryo fibroblasts (CEF) contains phosphorus linked to serine and threonine by monoester bonds. Normal and Rous sarcoma virus (RSV)-transformed cells were incubated with [32P]orthophosphate, and fibronectin was isolated from the cell surfaces and conditioned media. 32P was stably associated with fibronectin during immunoprecipitation, SDS-polyacrylamide gel electrophoresis, phospholipid solvent extraction, and hot acid but not alkaline treatment. After a limited acid hydrolysis of fibronectin, both phosphoserine and phosphothreonine were found. The specific radioactivity of the 32P-labeled fibronectin from the conditioned medium of normal CEF was higher than that from the cultures of transformed CEF. PMID:457771

  19. 10 CFR Appendix L to Part 110 - Illustrative List of Byproduct Materials Under NRC Export/Import Licensing Authority a

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... (Os 193) Oxygen 15 (O 15) Palladium 103 (Pd 103) Palladium 109 (Pd 109) Phosphorus 32 (P 32) Phosphorus 33 (P 33) Platinum 191 (Pt 191) Platinum 193m (Pt 193m) Platinum 193 (Pt 193) Platinum 197m (Pt...

  20. Orbital Rolls to Launch Pad at Wallops for Station Flight

    NASA Image and Video Library

    An Orbital Sciences Corporation Antares rolled out to launch Pad-0A at NASA's Wallops Flight Facility, Sunday, January 5, 2014, in advance of a planned Wednesday, Jan. 8th, 1:32 p.m. EST launch. Th...

  1. Phosphorus cycling and partitioning in an oligotrophic Everglades wetland ecosystem: A radioisotope tracing study

    USGS Publications Warehouse

    Noe, G.B.; Scinto, L.J.; Taylor, J.; Childers, D.L.; Jones, R.D.

    2003-01-01

    1. Our goal was to quantify short-term phosphorus (P) partitioning and identify the ecosystem components important to P cycling in wetland ecosystems. To do this, we added P radiotracer to oligotrophic, P-limited Everglades marshes. 32PO4 was added to the water column in six 1-m2 enclosed mesocosms located in long-hydroperiod marshes of Shark River Slough, Everglades National Park. Ecosystem components were then repeatedly sampled over 18 days. 2. Water column particulates (>0.45 ??m) incorporated radiotracer within the first minute after dosing and stored 95-99% of total water column 32P activity throughout the study. Soluble (<0.45 ??m) 32P in the water column, in contrast, was always <5% of the 32P in surface water. Periphyton, both floating and attached to emergent macrophytes, had the highest specific activity of 32P (Bq g-131P) among the different ecosystem components. Fish and aquatic macroinvertebrates also had high affinity for P, whereas emergent macrophytes, soil and flocculent detrital organic matter (floc) had the lowest specific activities of radiotracer. 3. Within the calcareous, floating periphyton mats, 81% of the initial 32P uptake was associated with Ca, but most of this 32P entered and remained within the organic pool (Ca-associated = 14% of total) after 1 day. In the floc layer, 32P rapidly entered the microbial pool and the labile fraction was negligible for most of the study. 4. Budgeting of the radiotracer indicated that 32P moved from particulates in the water column to periphyton and floc and then to the floc and soil over the course of the 18 days incubations. Floc (35% of total) and soil (27%) dominated 32P storage after 18 days, with floating periphyton (12%) and surface water (10%) holding smaller proportions of total ecosystem 32P. 5. To summarise, oligotrophic Everglades marshes exhibited rapid uptake and retention of labile 32P. Components dominated by microbes appear to control short-term P cycling in this oligotrophic ecosystem.

  2. Mechanism of the mRNA guanylyltransferase reaction: isolation of N epsilon-phospholysine and GMP (5' leads to N epsilon) lysine from the guanylyl-enzyme intermediate.

    PubMed Central

    Toyama, R; Mizumoto, K; Nakahara, Y; Tatsuno, T; Kaziro, Y

    1983-01-01

    The mRNA capping reaction catalyzed by rat liver mRNA guanylyltransferase proceeds through an enzyme-GMP intermediate in which GMP is linked to the enzyme by a phosphoamide linkage. The studies described here show that GMP is bound to the epsilon-amino group of lysine of rat liver guanylyltransferase. The enzyme-[32P]GMP intermediate was digested with pronase to a [32P]GMP-peptide which was then converted to [32P]phosphoryl-peptide through periodate oxidation followed by beta-elimination. After alkaline hydrolysis of the [32P]phosphoryl-peptide, the major radioactive product co-electrophoresed with the authentic N epsilon-phospholysine on DEAE-cellulose paper. Neither [32P]Nimid-phosphohistidine nor Nguanido-phosphoarginine was detected in the hydrolysates. Furthermore, formation of N epsilon-guanylyl-lysine linkage on the enzyme was more directly shown by isolation of [32P]GMP(5' leads to N epsilon)lysine when the steps of periodate oxidation and beta-elimination were omitted. The results indicate that the nucleophile in the guanylyltransferase to which the guanylyl residue is linked is the epsilon-amino group of a lysine residue. [32P]Phosphoryl-lysine was also isolated from the vaccinia virus capping enzyme-[32P]GMP intermediate. Guanylyltransferase from HeLa cells, wheat germ, Artemia salina and yeast also formed the enzyme-GMP complex and, from the stability of the complex, the linkage between the enzyme and GMP was suggested to be a phosphoamide. Images Fig. 2. Fig. 3. Fig. 4. Fig. 6. Fig. 7. Fig. 8. PMID:6321153

  3. 32-Phosphorus selectively delivered by listeria to pancreatic cancer demonstrates a strong therapeutic effect.

    PubMed

    Chandra, Dinesh; Selvanesan, Benson Chellakkan; Yuan, Ziqiang; Libutti, Steven K; Koba, Wade; Beck, Amanda; Zhu, Kun; Casadevall, Arturo; Dadachova, Ekaterina; Gravekamp, Claudia

    2017-02-06

    Our laboratory has developed a novel delivery platform using an attenuated non-toxic and non-pathogenic bacterium Listeria monocytogenes that infects tumor cells and selectively survives and multiplies in metastases and primary tumors with help of myeloid-derived suppressor cells (MDSC) and immune suppression in the tumor microenvironment (TME). 32P was efficiently incorporated into the Listeria bacteria by starvation of the bacteria in saline, and then cultured in phosphorus-free medium complemented with 32P as a nutrient. Listeria-32P kills tumor cells through both 32P-induced ionizing radiation and Listeria-induced reactive oxygen species (ROS). The levels of 32P and Listeria were studied in various normal and tumor tissues, at sequential time points after injection of mice with pancreatic cancer (syngeneic model Panc-02). We found that 32P and Listeria predominantly accumulated in tumors and metastases, with their highest accumulation 4 hrs (32P) and 3 days (Listeria) after injection. Listeria also penetrated the transgenic KPC (conditionally express endogenous Kras-G12D and p53-R172H mutant alleles) pancreatic tumors and metastases. This is remarkable since KPC tumors, like human tumors, exhibit a stromal barrier, which prevents most drugs from penetrating the pancreatic tumors. Therapeutic treatment with Listeria -32P resulted in a strong reduction of the growth of pancreatic cancer at early and late stages in Panc-02 and KPC mice. These results highlight the power of Listeria as new delivery platform of anticancer agents to the TME. Not only were therapeutic levels of radioactive Listeria reached in tumors and metastases but the selective delivery also led to minimal side effects.

  4. Phospholipid turnover and ultrastructural correlates during spontaneous germinal vesicle breakdown of the bovine oocyte: Effects of a cyclic AMP phosphodiesterase inhibitor

    SciTech Connect

    Homa, S.T.; Webster, S.D.; Russell, R.K. )

    1991-08-01

    The turnover of (32P)orthophosphate in bovine oocyte phospholipids was studied during the early stages of spontaneous meiotic maturation, and during inhibition of this process by the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX). Radioactive lipids were separated by TLC and the meiotic stage was determined cytogenetically. Ultrastructure of the nuclear membrane was examined using transmission EM. During the commitment period to meiotic resumption, which precedes germinal vesicle breakdown (GVBD), small localized convolutions appeared in the intact nuclear membrane. This was accompanied by a decrease in (32P)phosphatidic acid (PA) and an increase in (32P)-phosphatidylcholine (PC). This was followed by extensive convolutions, and subsequent dissociation, of the nuclear membrane, concomitant with a tremendous surge in (32P)PC and (32P)phosphatidylethanolamine (PE). The cAMP-mediated maintenance of meiotic arrest involved retention of entire nuclear envelope integrity and total inhibition of the surge in (32P)PC and (32P)PE which accompanied GVBD. The increase in (32P)phosphatidylinositol (PI) associated with all stages of early meiotic resumption was unaffected by IBMX. Microinjection of heparin inhibited GVBD, and injection of inositol 1,4,5-trisphosphate (IP3) overrode IBMX-maintained meiotic arrest in almost 40% of the oocytes. The results suggest that there may be several functions for phospholipid turnover in the regulation of spontaneous meiotic resumption in the bovine oocyte. The first precedes the commitment period, and involves IP3 generation to serve as the primary signal for meiotic resumption. The second occurs concomitant with the commitment period, is unaffected by the level of intracellular cAMP, and is associated with the general turnover of phospholipid.

  5. Phosphatidylinositol turnover in mitogen-activated lymphocytes. Suppression by low-density lipoproteins

    PubMed Central

    Hui, David Y.; Harmony, Judith A. K.

    1980-01-01

    Low-density (LD) lipoproteins inhibit phytohaemagglutinin-enhanced turnover of phosphatidylinositol in human peripheral lymphocytes. Turnover was assessed by 32P incorporation into phospholipids and by loss of 32P from [32P]phosphatidylinositol. Inhibition of lipid turnover by LD lipoproteins is not the result of a change in the amount of phytohaemagglutinin required for maximum cellular response. Neither phytohaemagglutinin nor LD lipoproteins influence 32P incorporation into phosphatidylethanolamine and phosphatidylcholine during the first 60min after mitogenic challenge. The extent of inhibition of phosphatidylinositol turnover by LD lipoproteins depends on the concentration of LD lipoproteins present in the incubation medium: 50% of maximum inhibition occurs at a low-density-lipoprotein protein concentration of 33μg/ml and maximum inhibition occurs at low-density-lipoprotein protein concentrations above 100μg/ml. Phytohaemagglutinin stimulates 32P incorporation into phosphatidylinositol, phosphatidylinositol phosphate and phosphatidylinositol bisphosphate. However, LD lipoproteins abolish 32P incorporation into phosphatidylinositol without affecting incorporation into phosphatidylinositol phosphate and phosphatidylinositol bisphosphate. The ability of LD lipoproteins to inhibit phytohaemagglutinin-induced phosphatidylinositol turnover is mimicked by EGTA. Furthermore, inhibition of LD lipoproteins by phytohaemagglutinin-induced 32P incorporation into phosphatidylinositol correlates directly with inhibition by LD lipoproteins of Ca2+ accumulation. These results suggest that Ca2+ accumulation and turnover of phosphatidylinositol are coupled responses in lymphocytes challenged by mitogens. The step in phosphatidylinositol metabolism that is sensitive to LD lipoproteins and, by inference, that is coupled to Ca2+ accumulation is release of [32P]phosphoinositol from phosphatidylinositol. PMID:6796039

  6. Platelet-activating factor stimulates metabolism of phosphoinositides in horse platelets: possible relationship to Ca2+ mobilization during stimulation.

    PubMed Central

    Billah, M M; Lapetina, E G

    1983-01-01

    Stimulation of horse platelets with platelet-activating factor (PAF) induces a rapid degradation of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Addition of 0.1 microM PAF for 5 sec to platelets prelabeled with 32P induces a 50% loss of [32P]PtdIns(4,5)P2. 32P-Labeled phosphatidylinositol 4-monophosphate (PtdIns4P) and [32P]phosphatidylinositol (PtdIns) also are decreased, albeit at a slower rate. Loss of 32P radioactivity correlates with a net loss of fatty acids from both polyphosphoinositides. Stimulation of platelets with PAF also produces formation of [32P]phosphatidic acid and [32P]lysophosphatidylinositol. The initial disappearance of inositol lipids is subsequently followed by resynthesis, as evidenced by increased incorporation of 32P into PtdIns(4,5)P2, PtdIns4P, and PtdIns. The resynthesis of the inositides increases with time and is proportional to the concentration of PAF. Prostacyclin (1 microM) inhibits (i) the formation of phosphatidic acid and lysophosphatidylinositol and (ii) the resynthesis of polyphosphoinositides induced by 0.03 microM PAF without affecting the initial loss of PtdIns(4,5)P2. The loss of inositol lipids appears to be a primary event of platelet activation. The initial loss of polyphosphoinositides might be linked to the initiation of cellular activation by mobilizing membrane-bound Ca2+, whereas the subsequent formation of these lipids might be involved in mechanisms to prevent overstimulation of the cell. PMID:6341992

  7. 32-Phosphorus selectively delivered by listeria to pancreatic cancer demonstrates a strong therapeutic effect

    PubMed Central

    Chandra, Dinesh; Selvanesan, Benson Chellakkan; Yuan, Ziqiang; Libutti, Steven K; Koba, Wade; Beck, Amanda; Zhu, Kun; Casadevall, Arturo; Dadachova, Ekaterina; Gravekamp, Claudia

    2017-01-01

    Our laboratory has developed a novel delivery platform using an attenuated non-toxic and non-pathogenic bacterium Listeria monocytogenes that infects tumor cells and selectively survives and multiplies in metastases and primary tumors with help of myeloid-derived suppressor cells (MDSC) and immune suppression in the tumor microenvironment (TME). 32P was efficiently incorporated into the Listeria bacteria by starvation of the bacteria in saline, and then cultured in phosphorus-free medium complemented with 32P as a nutrient. Listeria-32P kills tumor cells through both 32P-induced ionizing radiation and Listeria-induced reactive oxygen species (ROS). The levels of 32P and Listeria were studied in various normal and tumor tissues, at sequential time points after injection of mice with pancreatic cancer (syngeneic model Panc-02). We found that 32P and Listeria predominantly accumulated in tumors and metastases, with their highest accumulation 4 hrs (32P) and 3 days (Listeria) after injection. Listeria also penetrated the transgenic KPC (conditionally express endogenous Kras-G12D and p53-R172H mutant alleles) pancreatic tumors and metastases. This is remarkable since KPC tumors, like human tumors, exhibit a stromal barrier, which prevents most drugs from penetrating the pancreatic tumors. Therapeutic treatment with Listeria -32P resulted in a strong reduction of the growth of pancreatic cancer at early and late stages in Panc-02 and KPC mice. These results highlight the power of Listeria as new delivery platform of anticancer agents to the TME. Not only were therapeutic levels of radioactive Listeria reached in tumors and metastases but the selective delivery also led to minimal side effects. PMID:28186976

  8. Biosynthesis of rat enamel matrix components in vivo

    SciTech Connect

    Sasaki, S.; Shimokawa, H.; Tanaka, K.

    1982-12-01

    The biosynthesis of enamel matrix components of developing rat incisors was investigated by measuring the incorporation of /sup 3/H-proline, /sup 32/P-phosphate, and /sup 35/S-sulfate in vivo. /sup 3/H- and /sup 32/P-radioactivity was found in what seemed to be a prototype of enamel proteins. Subsequent shifts in other protein and peptide fractions were observed. /sup 35/S was also incorporated into components other than the above-mentioned proteins.

  9. Mode of Action of Shigella Toxin: Effects on Ribosome Structure and Function.

    DTIC Science & Technology

    1985-10-01

    at 40C/12hr. with [32P-5’]pCp, S.A.=2500 Ci/mmol, and T4 ligase as described by Peattie (38). The sample was precipitated with 70% ethanol and...32P-5’]pCp and T4 ligase , subjected to electrophoresis in urea-polyacrylamide gels and analyzed following autoradiography. The results indicate that

  10. Metabolic regulation during early frog development: flow of glycolytic carbon into phospholipids in Xenopus oocytes and fertilized eggs

    SciTech Connect

    Dworkin, M.B.; Dworkin-Rastl, E.

    1989-04-01

    /sup 32/P-labeled glucose 6-phosphate, (/sup 32/P)phosphoenolpyruvate, and (gamma-/sup 32/P)ATP were injected into oocytes and fertilized eggs of Xenopus laevis, and the incorporation of the /sup 32/P label was followed into phospholipids. Several classes of phospholipids incorporated /sup 32/P label from the injected glycolytic intermediates, including lysophosphatidic acid, phosphatidic acid, phosphatidylinositol, and phosphatidylinositol phosphates, inferring de novo synthesis of these lipids from dihydroxyacetone phosphate or glycerol 3-phosphate. Injection of (gamma-/sup 32/P)ATP into oocytes and fertilized eggs led to labeling of phosphatidylinositol phosphate and phosphatidylinositol bisphosphate, indicating an active phosphatidylinositol cycle in resting oocytes and fertilized eggs. Maturation and fertilization of the oocyte led to a qualitative change in phosphatidylinositol metabolism, increased labeling of phosphatidylinositol phosphate compared to phosphatidylinositol bisphosphate (either from glycerol 3-phosphate or from ATP). This change occurs late in the maturation process, and the new pattern of phosphatidylinositol metabolism is maintained during the rapid cleavage stages of early embryogenesis.

  11. The phosphorylated intermediate in the phosphoglyceromutase reaction

    PubMed Central

    Zwaig, N.; Milstein, C.

    1966-01-01

    1. High-voltage paper-electrophoresis methods have been used for the separation of 32P-labelled phosphoesters. 2. Evidence is presented which indicates that 32P-labelled phosphopeptides, obtained after acid hydrolysis of phosphoglyceromutase incubated with impure 2,3-di[32P]phosphoglycerate, are derived from phosphoglucomutase contamination. 3. The hydrolysis of 2,3-di[32P]phosphoglycerate by phosphoglyceromutase has been studied. After an apparent instantaneous hydrolysis of 1 mole of coenzyme/mole of enzyme the reaction proceeds at a very low rate. 4. This hydrolysis seems to be due to the destruction of an enzyme–coenzyme complex. The proportions of the decomposition products of the complex vary according to further handling (pH of ionophoresis). 5. The inorganic [32P]phosphate produced by the hydrolysis of the complex and the inorganic [32P]phosphate produced by the slow phosphatase activity can be differentiated by the ability of the former to be incorporated into non-radioactive substrate before enzyme denaturation. 6. The effect of enzyme concentration on the stoicheiometry of the slow phosphatase hydrolysis of the diphosphoglycerate is described and this suggests that it may occur via two independent reactions, one of them being the decomposition of the enzyme–coenzyme complex on standing. PMID:5941332

  12. Relationship between stimulated phosphatidic acid production and inositol lipid hydrolysis in intestinal longitudinal smooth muscle from guinea pig.

    PubMed

    Mallows, R S; Bolton, T B

    1987-06-15

    Accumulation of [32P]phosphatidic acid (PA) and total [3H]inositol phosphates (IPs) was measured in the longitudinal smooth-muscle layer from guinea-pig small intestine. Stimulation with carbachol, histamine and substance P produced increases in accumulation of both [3H]IPs and [32P]PA over the same concentration range. The increase in [32P]PA accumulation in response to carbachol (1 microM-0.1 mM) was inhibited in the presence of atropine (0.5 microM). Buffering the external free [Ca2+] to 10 nM did not prevent the carbachol-stimulated increase in [32P]PA accumulation. Carbachol and Ca2+ appear to act synergistically to increase accumulation of [32P]PA. In contrast, although incubation with noradrenaline also increased accumulation of [3H]IPs, no increase in accumulation of [32P]PA could be detected. These results suggest that an increase in formation of IPs is not necessarily accompanied by an increase in PA formation, and imply the existence of receptor-modulated pathways regulating PA concentrations other than by phospholipase-C-catalysed inositol phospholipid hydrolysis.

  13. Phosphate fluxes in single muscle fibres of the spider crab, Maia squinado.

    PubMed Central

    Caldwell, P C; Lowe, A G

    1980-01-01

    1. The rates of influx and efflux of [32P]orthophosphate (Pi) have bben measured in single muscle fibres of the spider crab, Maia squinado. 2. Rates of influx of 32Pi from salines containing 0 . 11 to 0 . 37 mM-Pi ranged from 0 . 11 to 0 . 41 p-mole cm-2 sec-1. 3. After injection of [32P]orthophosphate there was an early rapid phase of 32P efflux which was maximal after about 5-10 min, then a gradual decline until a low steady-state efflux rate was established after about 100 min. The rate of decline of 32P efflux was similar to the rate of equilibration of injected 32Pi with arginine phosphate and ATP. 4. The steady-state rate of 32P efflux was typically about 0 . 70 p-mole cm-2 sec-1 when the sarcoplasmic Pi concentraion was 5 m-mole kg-1. Exposure of mucle fibres to conditions which caused contraction and increased the sarcoplasmic concentration of Pi also increased the rate of 32P efflux, which appeared to be linearly related to sarcoplasmic Pi concentration. 5. The results are compared with previous measurements of phosphate fluxes in nerve. PMID:7411440

  14. Phosphate fluxes in single muscle fibres of the spider crab, Maia squinado.

    PubMed

    Caldwell, P C; Lowe, A G

    1980-04-01

    1. The rates of influx and efflux of [32P]orthophosphate (Pi) have bben measured in single muscle fibres of the spider crab, Maia squinado. 2. Rates of influx of 32Pi from salines containing 0 . 11 to 0 . 37 mM-Pi ranged from 0 . 11 to 0 . 41 p-mole cm-2 sec-1. 3. After injection of [32P]orthophosphate there was an early rapid phase of 32P efflux which was maximal after about 5-10 min, then a gradual decline until a low steady-state efflux rate was established after about 100 min. The rate of decline of 32P efflux was similar to the rate of equilibration of injected 32Pi with arginine phosphate and ATP. 4. The steady-state rate of 32P efflux was typically about 0 . 70 p-mole cm-2 sec-1 when the sarcoplasmic Pi concentraion was 5 m-mole kg-1. Exposure of mucle fibres to conditions which caused contraction and increased the sarcoplasmic concentration of Pi also increased the rate of 32P efflux, which appeared to be linearly related to sarcoplasmic Pi concentration. 5. The results are compared with previous measurements of phosphate fluxes in nerve.

  15. Influence of cyclic nucleotides (cAMP) on inositol phospholipid (InsPL) metabolism in cultured mesangial (MS) cells

    SciTech Connect

    Troyer, D.A.; Venkatachalam, M.A.; Bonventre, J.V.; Kreisberg, J.I.

    1986-03-01

    Elevation of cAMP inhibits hormone-induced contraction of MS cells, and in other cell types, reduces stimulated InsPL metabolism. The authors found that neither isobutylmethylxanthine (MIX, 0.5 mM), which increases MS cell cAMP levels 4-fold, nor forskolin (100 ..mu..M) altered vasopressin (VP, 10 nM) induced release of /sup 3/H-inositol trisphosphate from prelabelled MS cells. Also, maneuvers which elevated cAMP did not block the VP-induced rise of intracellular calcium as measured by quin-2. Further, neither MIX nor isoproterenol affected the stimulation of glycolysis by VP as measured by lactic acid production. MIX diminished VP stimulated /sup 32/P orthophosphate (/sup 32/P) incorporation into phosphatidylinositol 4,5-bisphosphate, phosphatidylinositol 4-phosphate and phosphatidylinositol. The /sup 32/P content in phosphoinositides of cells treated with MIX and VP was 65% of that in cells treated with VP only. However, the authors found that the specific activity of /sup 32/P in ATP in the presence of MIX + VP was 74% of that with VP alone. Thus, the apparent suppression of /sup 32/P incorporation due to MIX was attributable to a concurrent diminution of the specific activity of /sup 32/P in ATP. The authors conclude that increases of cAMP interfere with contraction distal to PIP/sub 2/ hydrolysis, inositol phosphate release, calcium mobilization, and enhancement of glycolysis.

  16. Role of zinc in the structure and function of ssDNA binding proteins

    SciTech Connect

    Coleman, J.E.; Giedroc, D.P.; Keating, K.M.; Williams, K.R.

    1987-05-01

    Gene 32 protein (g32P), the ssDNA binding protein required for replication, recombination and translational control in the phage T4 life cycle contains 1 g at Zn per mol bound in a tetrahedral complex to -S/sup -/ ligands. Chemical modification and spectroscopic evidence suggest binding to Cys 77, His 81, Cys 87 and Cys 90. The Zn-binding domain is at the N-terminal end of AA residues 72 to 116 which contains 6 of the 8 Tyr residues in g32P, shown by /sup 1/H-NMR to be involved in deoxynucleotide binding. Limited proteolysis of g32P with trypsin removes residues 1-21 and 254-301 leaving a trypsin-resistant core, g32P. The latter retains high affinity for a single site nucleotide lattice, but has lost cooperative binding to DNA. Removal of Zn from native g32P renders the molecule susceptible to proteolysis and the core is hydrolyzed to small peptides. Rebinding of Zn restores the core stability. Apo g32P binds 3 orders of magnitude less tightly to ssDNA and cannot melt polyd(AT) at 150 mM NaCl. Loss of binding affinity is primarily due to loss of cooperative protein-protein interactions accompanying Zn removal. Thus, both the N-terminal domain and the Zn-binding domain are required for cooperative binding to DNA. G5P from fd and ssB from E. coli do not contain Zn, but small basic proteins, products of the gag gene of retroviruses, e.g., p10 from MuLV, p15 from HTLVI and p15 from HTLV-III contain tandem repeats of a -C-X/sub 2/-C-X/sub 4/-H-X/sub 4/-C- sequence similar to the Zn-binding sequence found in g32P.

  17. [Phosphorus translocation and distribution in intercropping systems of soybean (Glycine max) and citrus (Citrus poonensis)].

    PubMed

    Zhou, Weijun; Wang, Kairong; Li, Hesong

    2004-02-01

    A field mini-plot experiment was conducted on clay loamy oxisol using 32P trace technique when P fertilizer was applied in three depth soil (15, 35 and 55 cm soil layer) to compare P absorption, distribution and translocation in plant organ and soil profile under soybean and citrus monoculture and intercropping at Taoyuan Experimental Station of Agroecosystem Research of Chinese Academy of Science. Total P uptake (PT) and P accumulation in different parts (PA) of soybean were remarkably decreased under intercropping. When 32P was applied in topsoil (15 cm soil layer), 32P uptake (32PT) by soybean was significantly lower in intercropping than in monoculture. Whereas 32PT uptake by soybean was significantly greater in intercropping than in monoculture when 32P was applied in deep soil layer (35 cm or 55 cm soil layer). However, considerable difference was not observed for 32P translocation and distribution among soybean organs. 32PT uptake by citrus was much lower under intercropping than under monoculture. The P uptake by citrus newly could be transferred rapidly to aboveground and prior to active growing organ. Intercropping did not affect 32P distribution in citrus organ, but when P was applied in deep soil layer, the speed of 32P transferred to aboveground and active organ was slowed down. P mobility was strengthened in soil profile, and P of deep soil layer was promoted to move to topsoil in intercropping. The experimental results showed the optimal depth of applied P should be within 20 cm soil layer in soybean-citrus intercropping system.

  18. Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase.

    PubMed

    Arisz, Steven A; van Wijk, Ringo; Roels, Wendy; Zhu, Jian-Kang; Haring, Michel A; Munnik, Teun

    2013-01-01

    Phosphatidic acid (PtdOH) is emerging as an important signaling lipid in abiotic stress responses in plants. The effect of cold stress was monitored using (32)P-labeled seedlings and leaf discs of Arabidopsis thaliana. Low, non-freezing temperatures were found to trigger a very rapid (32)P-PtdOH increase, peaking within 2 and 5 min, respectively. In principle, PtdOH can be generated through three different pathways, i.e., (1) via de novo phospholipid biosynthesis (through acylation of lyso-PtdOH), (2) via phospholipase D hydrolysis of structural phospholipids, or (3) via phosphorylation of diacylglycerol (DAG) by DAG kinase (DGK). Using a differential (32)P-labeling protocol and a PLD-transphosphatidylation assay, evidence is provided that the rapid (32)P-PtdOH response was primarily generated through DGK. A simultaneous decrease in the levels of (32)P-PtdInsP, correlating in time, temperature dependency, and magnitude with the increase in (32)P-PtdOH, suggested that a PtdInsP-hydrolyzing PLC generated the DAG in this reaction. Testing T-DNA insertion lines available for the seven DGK genes, revealed no clear changes in (32)P-PtdOH responses, suggesting functional redundancy. Similarly, known cold-stress mutants were analyzed to investigate whether the PtdOH response acted downstream of the respective gene products. The hos1, los1, and fry1 mutants were found to exhibit normal PtdOH responses. Slight changes were found for ice1, snow1, and the overexpression line Super-ICE1, however, this was not cold-specific and likely due to pleiotropic effects. A tentative model illustrating direct cold effects on phospholipid metabolism is presented.

  19. Potential use of P-32 ophthalmic applicator: Monte Carlo simulations for design and dosimetry

    SciTech Connect

    Park, Yang Kyun; Ye, Sung-Joon; Kim, Il Han; Wee, Won Ryang; Kim, Mee Kum; Han, Hyon Soo; Son, Kwang-Jae; Park, Ul Jae

    2008-05-15

    Postoperative {beta}-irradiation after pterygium excision has been considered a valuable therapeutic procedure to reduce the recurrence rate. Recently, it was reported that {beta}-irradiation also substantially reduced the risk of surgical failure after glaucoma surgery. Pure {beta}-irradiation using a {sup 90}Sr/Y applicator has been almost exclusively used for this purpose. As an alternative to {sup 90}Sr/Y {beta}-irradiation, we propose treatment with betas of a {sup 32}P source. While {sup 32}P has a lower maximum energy (1.71 MeV) than {sup 90}Sr/Y (2.27 MeV), it has an average energy comparable to that of {sup 90}Sr/Y. Furthermore, it can be produced easily in a nuclear reactor by neutron activation and is considered a less hazardous material. Monte Carlo simulations for the dosimetry of proposed {sup 32}P applicators were performed using the MCNP5 code. The structure and dimension of the {sup 32}P applicators were based on those of the {sup 90}Sr/Y applicators currently available, while medical plastic encapsulation and liquid source were chosen to enhance {beta}-dose to the surface of the conjunctiva. The {sup 32}P applicator showed that the surface dose distribution (up to 0.75 mm depth) is very similar to that of {sup 90}Sr/Y. However, beyond 0.75 mm depth, the {sup 32}P doses decrease with depths more rapidly than {sup 90}Sr/Y doses. In order to achieve the same surface dose rate, the required {sup 32}P activity is about three times that for a {sup 90}Sr/Y applicator. We conclude that the proposed {sup 32}P applicator can deliver therapeutic doses to the target lesion while sparing the lens better than the {sup 90}Sr/Y applicator. The {sup 32}P activity required to deliver therapeutic doses can be produced in a 30 MW reactor available at the Korea Atomic Energy Research Institute.

  20. The energy spectrum of neutrons from 7Li(d,n)8Be reaction at deuteron energy 2.9 MeV

    NASA Astrophysics Data System (ADS)

    Mitrofanov, Konstantin V.; Piksaikin, Vladimir M.; Zolotarev, Konstantin I.; Egorov, Andrey S.; Gremyachkin, Dmitrii E.

    2017-09-01

    The neutron beams generated at the electrostatic accelerators using nuclear reactions T(p,n)3He, D(d,n)3He, 7Li(p,n)7Be, T(d,n)4He, 7Li(d,n)8Be, 9Be(d,n)10B are widely used in neutron physics and in many practical applications. Among these reactions the least studied reactions are 7Li(d,n)8Be and 9Be(d,n)10B. The present work is devoted to the measurement of the neutron spectrum from 7Li(d,n)8Be reaction at 0∘ angle to the deuteron beam axis on the electrostatic accelerator Tandetron (JSC "SSC RF - IPPE") using activation method and a stilbene crystal scintillation detector. The first time ever 7Li(d,n)8Be reaction was measured by activation method. The target was a thick lithium layer on metallic backing. The energy of the incident deuteron was 2.9 MeV. As activation detectors a wide range of nuclear reactions were used: 27Al(n,p)27Mg, 27Al(n,α)24Na, 113In(n,n')113mIn, 115In(n,n')115mIn, 115In(n,γ)116mIn, 58Ni(n,p)58mCo, 58Ni(n,2n)57Ni, 197Au(n,γ)198Au, 197Au(n,2n)196Au, 59Co(n,p)59Fe, 59Co(n,2n)58m+gCo, 59Co (n,g)60Co. Measurement of the induced gamma-activity was carried out using HPGe detector Canberra GX5019 [1]. The up-to-date evaluations of the cross sections for these reactions were used in processing of the data. The program STAYSL was used to unfold the energy spectra. The neutron spectra obtained by activation detectors is consistent with the corresponding data measured by a stilbene crystal scintillation detector within their uncertainties.

  1. PROTOPLANETARY DISK STRUCTURES IN OPHIUCHUS. II. EXTENSION TO FAINTER SOURCES

    SciTech Connect

    Andrews, Sean M.; Wilner, D. J.; Hughes, A. M.; Qi, Chunhua; Dullemond, C. P.

    2010-11-10

    We present new results from a significant extension of our previous high angular resolution (0.''3 {approx} 40 AU) submillimeter array survey of the 340 GHz (880 {mu}m) thermal continuum emission from dusty circumstellar disks in the {approx}1 Myr old Ophiuchus star-forming region. An expanded sample is constructed to probe disk structures that emit significantly lower millimeter luminosities (hence dust masses), down to the median value for T Tauri stars. Using a Monte Carlo radiative transfer code, the millimeter visibilities and broadband spectral energy distribution for each disk are simultaneously reproduced with a two-dimensional parametric model for a viscous accretion disk that has a surface density profile {Sigma} {proportional_to} (R/R{sub c}){sup -}{gamma}exp [- (R/R{sub c}){sup 2-}{gamma}]. We find wide ranges of characteristic radii (R{sub c} = 14-198 AU) and disk masses (M{sub d} = 0.004-0.143 M{sub sun}), but a narrow distribution of surface density gradients ({gamma} = 0.4-1.1) that is consistent with a uniform value ({gamma}) = 0.9 {+-} 0.2 and independent of mass (or millimeter luminosity). In this sample, we find a correlation between the disk luminosity/mass and characteristic radius, such that fainter disks are both smaller and less massive. We suggest that this relationship is an imprint of the initial conditions inherited by the disks at their formation epoch, compare their angular momenta with those of molecular cloud cores, and speculate on how future observations can help constrain the distribution of viscous evolution timescales. No other correlations between disk and star properties are found. The inferred disk structures are briefly compared with theoretical models for giant planet formation, although resolution limitations do not permit us to directly comment on material inside R {approx} 20 AU. However, there is some compelling evidence for the evolution of dust in the planet formation region: 4/17 disks in the sample show resolved

  2. [Eye and lymph drainage].

    PubMed

    Grüntzig, J; Schicha, H; Huth, F

    1979-06-01

    Up to now lymphatics in the eye could not be pointed out. An ocular lymph drainage is denied. Földi succeeded in producing experimentally the syndrome of "lymphostatic encephalopathy and ophthalmopathy" by operative blockade of the cervical lymphatics in animals. In the first part of the present paper a historical view considering the subject "Eye and lymphatic system" is given. In the second part it is entered into the particulars of own experimental studies. As to our own investigations, rabbits have been injected 99mTc-sulfur-colloid, 99mTc-microcolloid, 99mTc-Albumin and 198Au-colloid into the retrobulbar space, anterior chamber, vitreous body and subconjuctival space of one eye. Measurements of the activity's distribution have been made in vivo with an Anger type camera (pho-Gamma-IV Hp, Searle Nuclear Chicago) and in vitro after section with a sodium iodine crystal well counter (Clinimat-200, Picker). In some animals the investigation has been combined with a bilateral dissection of the cervical lymph nodes. After injection in the retrobulbar space a significant concentration of the activity could be observed for the most part in the equilateral Lymphonodulus cervicalis profundus. By the cervical lymph blockade the removal of lymphoctopic substances from the retrobulbar space was largely inhibited. After injection in the anterior chamber a significant concentration could be observed for the most part in the equilateral Lymphonodulus cervicalis superficialis. After intravitreal injection a drainage to the bilateral deep cervical lymph nodes could be observed. After injection into the subconjunctival space a significant accumulation of activity could be registered in the equilateral Lymphonoduli mandibulares and cervicales superficiales. The data substantiate a segmental lymph drainage from the eye: vitreous body and retrobulbar space for the most part into the Lymphonoduli cervicales profundi, anterior chamber and subconjunctival space for the most part into

  3. Protein Phosphorylation during Coconut Zygotic Embryo Development1

    PubMed Central

    Islas-Flores, Ignacio; Oropeza, Carlos; Hernández-Sotomayor, S.M. Teresa

    1998-01-01

    Evidence was obtained on the occurrence of protein threonine, serine, and tyrosine (Tyr) kinases in developing coconut (Cocos nucifera L.) zygotic embryos, based on in vitro phosphorylation of proteins in the presence of [γ-32P]ATP, alkaline treatment, and thin-layer chromatography analysis, which showed the presence of [32P]phosphoserine, [32P]phosphothreonine, and [32P]phosphotyrosine in [32P]-labeled protein hydrolyzates. Tyr kinase activity was further confirmed in extracts of embryos at different stages of development using antiphosphotyrosine monoclonal antibodies and the synthetic peptide derived from the amino acid sequence surrounding the phosphorylation site in pp60src (RR-SRC), which is specific for Tyr kinases. Anti-phosphotyrosine western blotting revealed a changing profile of Tyr-phosphorylated proteins during embryo development. Tyr kinase activity, as assayed using RR-SRC, also changed during embryo development, showing two peaks of activity, one during early and another during late embryo development. In addition, the use of genistein, a Tyr kinase inhibitor, diminished the ability of extracts to phosphorylate RR-SRC. Results presented here show the occurrence of threonine, serine, and Tyr kinases in developing coconut zygotic embryos, and suggest that protein phosphorylation, and the possible inference of Tyr phosphorylation in particular, may play a role in the coordination of the development of embryos in this species. PMID:9733545

  4. Field and laboratory investigations of inactivation of viruses (PRD1 and MS2) attached to iron oxide-coated quartz san

    USGS Publications Warehouse

    Ryan, Joseph N.; Harvey, Ronald W.; Metge, David W.; Elimelech, Menachem; Navigato, Theresa; Pieper, Ann P.

    2002-01-01

    Field and laboratory experiments were conducted to investigate inactivation of viruses attached to mineral surfaces. In a natural gradient transport field experiment, bacteriophage PRD1, radiolabeled with 32P, was injected into a ferric oxyhydroxide-coated sand aquifer with bromide and linear alkylbenzene sulfonates. In a zone of the aquifer contaminated by secondary sewage infiltration, small fractions of infective and 32P-labeled PRD1 broke through with the bromide tracer, followed by the slow release of 84% of the 32P activity and only 0.011% of the infective PRD1. In the laboratory experiments, the inactivation of PRD1, labeled with 35S (protein capsid), and MS2, dual radiolabeled with 35S (protein capsid) and 32P (nucleic acid), was monitored in the presence of groundwater and sediment from the contaminated zone of the field site. Release of infective viruses decreased at a much faster rate than release of the radiolabels, indicating that attached viruses were undergoing surface inactivation. Disparities between 32P and35S release suggest that the inactivated viruses were released in a disintegrated state. Comparison of estimated solution and surface inactivation rates indicates solution inactivation is ∼3 times as fast as surface inactivation. The actual rate of surface inactivation may be substantially underestimated owing to slow release of inactivated viruses.

  5. Effect of mercuric chloride feeding on sexual maturity, egg production and fertility in Japanese quail

    USGS Publications Warehouse

    Hill, E.F.; Shaffner, C.S.

    1973-01-01

    Japanese quail (Coturnix c. japonica) were fed 0, 8, 16 or 32 p.p.m. of mercury as mercuric chloride from 3 days of age through 20 weeks of age. The onset of egg production generally occurred earlier for hens fed HgCl2. Average age in days at first oviposition for the control, 8 p.p.m., 16 p.p.m. and 32 p.p.m. was 48.4, 50.9, 46.9 and 44.0 respectively. The average rate of egg productivity from first oviposition to attainment of full growth (9 weeks of age) correlated positively with in increased dietary mercury (controls, 8 p.p.m., 16 p.p.m., 32 p.p.m. ? 75.2, 69.3, 86.1 and 93.3% respectively). By 20 weeks of age productivity was 81.0, 80.6, 87.5 and 92.9% for control, 8, 16 and 32 p.p.m. groups respectively. Fertility was depressed when hens were fed HgCl2. At 9 weeks of age average control fertility was 59% contrasted with 25% for the 32 p.p.m. group. At 12 weeks fertility increased to 89% and 57% for these groups. From this study it is apparent. that the onset and rate of egg production was stimulated by HgCl2, but fertility was adversely affected.

  6. Phosphorus-32, a Clinically Available Drug, Inhibits Cancer Growth by Inducing DNA Double-Strand Breakage

    PubMed Central

    Cheng, Yulan; Kiess, Ana P.; Herman, Joseph M.; Pomper, Martin G.; Meltzer, Stephen J.; Abraham, John M.

    2015-01-01

    Radioisotopes that emit electrons (beta particles), such as radioiodine, can effectively kill target cells, including cancer cells. Aqueous 32P[PO4] is a pure beta-emitter that has been used for several decades to treat non-malignant human myeloproliferative diseases. 32P[PO4] was directly compared to a more powerful pure beta-emitter, the clinically important 90Y isotope. In vitro, 32P[PO4] was more effective at killing cells than was the more powerful isotope 90Y (P ≤ 0.001) and also caused substantially more double-stranded DNA breaks than did 90Y. In vivo, a single low-dose intravenous dose of aqueous elemental 32P significantly inhibited tumor growth in the syngeneic murine cancer model (P ≤ 0.001). This effect is exerted by direct incorporation into nascent DNA chains, resulting in double-stranded breakage, a unique mechanism not duplicatable by other, more powerful electron-emitting radioisotopes. 32P[PO4] should be considered for human clinical trials as a potential novel anti-cancer drug. PMID:26030880

  7. Phosphorus-32 absorption and translocation to host plants by arbuscular mycorrhizal fungi at low root-zone temperature.

    PubMed

    Wang, B; Funakoshi, D M; Dalpé, Y; Hamel, C

    2002-04-01

    Arbuscular mycorrhizal (AM) mycelia persist in soil over winter. Functioning of the AM symbiosis very early in the spring when the soil temperature is low may be of ecological significance for perennial and biannual plants in cool climates. An indoor experiment was conducted to investigate the effects of low root-zone temperatures on 32P uptake by 10-week-old leek plants (Allium porrum L.) inoculated or not with the AM fungus Glomus intraradices Schenck & Smith. Plants were grown in a greenhouse at approximately 23 degrees C prior to exposing their roots to 23 degrees C, 15 degrees C or 0 degree C. Mycorrhizal colonization increased 32P activity of leek leaves at a root-zone temperature of 23 degrees C seven days after injection of 32P into the soil, whereas 14 days after injection, 32P increases were measured at both 23 degrees C and 15 degrees C. The lack of difference in 32P activity between AM and non-AM plants at 0 degree C, both 7 and 14 days after injection, suggests that the AM fungus is not functional at this low root-zone temperature.

  8. Acetylglyceryl ether phosphorylcholine (AGEPC; platelet-activating factor)-induced stimulation of rabbit platelets: correlation between phosphatidic acid level, 45Ca2+ uptake, and (3H)serotonin secretion

    SciTech Connect

    Shukla, S.D.; Hanahan, D.J.

    1984-08-01

    When 32P-labeled rabbits platelet were incubated with 5 X 10(-10) M 1-O-alkyl-2-acetyl-sn-glyceryl-3-phosphorylcholine (AGEPC), either in the presence or absence (0.1 mM EGTA) of added Ca2+, there was a three- to five-fold increase in the (32P)phosphatidic acid (PA) pool within 15 to 20 s. This event was followed by a gradual decrease in the (32P)PA level to near basal level in 5 min. AGEPC effected this change in (32P)PA in a characteristic dose- and time-dependent manner. Polar head group analogs of AGEPC, such as AGEDME and AGEMME, also effected an increase in PA labeling at levels comparable to those previously reported for their activity toward rabbit platelets. Other analogs, i.e., lysoGEPC and the enantiomer, sn-1-AGEPC, which are inactive toward rabbit platelets, also showed no effect on the level of (32P)PA. The finding that the PA level in rabbit platelets could be manipulated by the addition of AGEPC, without any added Ca2+, provided an excellent model system for establishing a correlation between the uptake of Ca2+, serotonin release, and PA level. Thus, PA must be regarded as a sensitive indicator of a reaction mechanism important to the platelet response to AGEPC, and could be the focal point in promoting calcium uptake during the stimulation process.

  9. Okadaic acid-induced inhibition of B-50 dephosphorylation by presynaptic membrane-associated protein phosphatases.

    PubMed

    Han, Y F; Dokas, L A

    1991-10-01

    The neuronal tissue-specific protein kinase C (PKC) substrate B-50 can be dephosphorylated by endogenous protein phosphatases (PPs) in synaptic plasma membranes (SPMs). The present study characterizes membrane-associated B-50 phosphatase activity by using okadaic acid (OA) and purified 32P-labeled substrates. At a low concentration of [gamma-32P]ATP, PKC-mediated [32P]phosphate incorporation into B-50 in SPMs reached a maximal value at 30 s, followed by dephosphorylation. OA, added 30 s after the initiation of phosphorylation, partially prevented the dephosphorylation of B-50 at 2 nM, a dose that inhibits PP-2A. At the higher concentration of 1 microM, a dose of OA that inhibits PP-1 as well as PP-2A, a nearly complete blockade of B-50 dephosphorylation was seen. Heat-stable PP inhibitor-2 (I-2) also inhibited dephosphorylation of B-50. The effects of OA and I-2 on B-50 phosphatase activity were additive. Endogenous PP-1- and PP-2A-like activities in SPMs were also demonstrated by their capabilities of dephosphorylating [32P]phosphorylase a and [32P]casein. With these exogenous substrates, sensitivities of the membrane-bound phosphatases to OA and I-2 were found to be similar to those of purified forms of these enzymes. These results indicate that PP-1- and PP-2A-like enzymes are the major B-50 phosphatases in SPMs.

  10. Phosphate dynamics in an acidic mountain stream: Interactions involving algal uptake, sorption by iron oxide, and photoreduction

    USGS Publications Warehouse

    Tate, Cathy M.; Broshears, Robert E.; McKnight, Diane M.

    1995-01-01

    Acid mine drainage streams in the Rocky Mountains typically have few algal species and abundant iron oxide deposits which can sorb phosphate. An instream injection of radiolabeled phosphate (32P0,) into St. Kevin Gulch, an acid mine drainage stream, was used to test the ability of a dominant algal species, Ulothrix sp., to rapidly assimilate phosphate. Approximately 90% of the injected phosphate was removed from the water column in the 175-m stream reach. When shaded stream reaches were exposed to full sunlight after the injection ended, photoreductive dissolution of iron oxide released sorbed 32P, which was then also removed downstream. The removal from the stream was modeled as a first-order process by using a reactive solute transport transient storage model. Concentrations of 32P mass-’ of algae were typically lo-fold greater than concentrations in hydrous iron oxides. During the injection, concentrations of 32P increased in the cellular P pool containing soluble, low-molecular-weight compounds and confirmed direct algal uptake of 32P0, from water. Mass balance calculations indicated that algal uptake and sorption on iron oxides were significant in removing phosphate. We conclude that in stream ecosystems, PO, sorbed by iron oxides can act as a dynamic nutrient reservoir regulated by photoreduction.

  11. Protein phosphorylation: Localization in regenerating optic axons

    SciTech Connect

    Larrivee, D. )

    1990-09-01

    A number of axonal proteins display changes in phosphorylation during goldfish optic nerve regeneration. (1) To determine whether the phosphorylation of these proteins was closely linked to their synthesis in the retinal ganglion cell body, cycloheximide was injected intraocularly into goldfish whose optic nerves had been regenerating for 3 weeks. Cycloheximide reduced the incorporation of (3H)proline and 32P orthophosphate into total nerve protein by 84% and 46%, respectively. Of the 20 individual proteins examined, 17 contained less than 15% of the (3H)proline label measured in corresponding controls, whereas 18 proteins contained 50% or more of the 32P label, suggesting that phosphorylation was largely independent of synthesis. (2) To determine whether the proteins were phosphorylated in the ganglion cell axons, axonal transport of proteins was blocked by intraocular injection of vincristine. Vincristine reduced (3H)proline labeling of total protein by 88% and 32P labeling by 49%. Among the individual proteins (3H)proline labeling was reduced by 90% or more in 18 cases but 32P labeling was reduced only by 50% or less. (3) When 32P was injected into the cranial cavity near the ends of the optic axons, all of the phosphoproteins were labeled more intensely in the optic tract than in the optic nerve. These results suggest that most of the major phosphoproteins that undergo changes in phosphorylation in the course of regeneration are phosphorylated in the optic axons.

  12. EFFECT OF DMPS AND DMSA ON THE PLACENTAL AND FETAL DISPOSITION OF METHYLMERCURY

    PubMed Central

    Bridges, Christy C.; Joshee, Lucy; Zalups, Rudolfs K.

    2009-01-01

    Methylmercury (CH3Hg+) is a serious environmental toxicant. Exposure to this metal during pregnancy can cause serious neurological and developmental defects in a developing fetus. Surprisingly, little is known about the mechanisms by which mercuric ions are transported across the placenta. Although it has been shown that 2,3-dimercaptopropane-1-sulfonate (DMPS) and 2,3-dimercaptosuccinic acid (DMSA) are capable of extracting mercuric ions from various organs and cells, there is no evidence that they are able to extract mercury from placental or fetal tissues following maternal exposure to CH3Hg+. Therefore, the purpose of the current study was to evaluate the ability of DMPS and DMSA to extract mercuric ions from placental and fetal tissues following maternal exposure to CH3Hg+. Pregnant Wistar rats were exposed to CH3HgCl, containing [203Hg], on day 11 or day 17 of pregnancy and treated 24 h later with saline, DMPS or DMSA. Maternal organs, fetuses, and placentas were harvested 48 h after exposure to CH3HgCl. The disposition of mercuric ions in maternal organs and tissues was similar to that reported previously by our laboratory. The disposition of mercuric ions in placentas and fetuses appeared to be dependent upon the gestational age of the fetus. The fetal and placental burden of mercury increased as fetal age increased and was reduced by DMPS and DMSA, with DMPS being more effective. The disposition of mercury was examined in liver, total renal mass, and brain of fetuses harvested on gestational day 19. On a per gram tissue basis, the greatest amount of mercury was detected in the total renal mass of the fetus, followed by brain and liver. DMPS and DMSA reduced the burden of mercury in liver and brain while only DMPS was effective in the total renal mass. The results of the current study are the first to show that DMPS and DMSA are capable of extracting mercuric ions, not only from maternal tissues, but also from placental and fetal tissues following maternal

  13. Mercury abundances and isotopic compositions in the Murchison (CM) and Allende (CV) carbonaceous chondrites

    NASA Astrophysics Data System (ADS)

    Lauretta, Dante S.; Klaue, Bjoern; Blum, Joel D.; Buseck, Peter R.

    2001-08-01

    suggests that Hg is associated with S-bearing phases and, thus, that HgS is a major Hg-bearing phase in both meteorites. The Hg peak at 225°C for Allende is similar to release patterns of physically adsorbed Hg on silicate and metal grains. Prior studies suggested that the isotopic anomalies reported from NAA resulted from interference between 203Hg and 75Se. However, the amount of Se released from both meteorites, relative to Hg, is insufficient to produce all of the observed anomalies.

  14. Mercury Benthic Flux: A Comparison Between 3 Mining-Impacted Water Bodies in the Western United States

    NASA Astrophysics Data System (ADS)

    Topping, B. R.; Kuwabara, J. S.; Marvin-Dipasquale, M. C.; Agee, J. L.; Kieu, L. H.; Flanders, J. R.; Parchaso, F.

    2004-12-01

    The legacy of mining in the Western United States has left an indelible environmental imprint on terrestrial and aquatic systems. On both sides of the Sierra Nevada mountain range (Sierras), mercury was used copiously in the amalgamation of gold and silver. Mercury deposits in close proximity to San Francisco Bay (e.g., the New Almaden mining district) provided much of the mercury for these processes. To evaluate mercury benthic flux, three geographically distinct water bodies were studied: Lahontan Reservoir (NV) on the eastern side of the Sierras, affected by historic gold and silver mining; Camp Far West Reservoir (CA) on the western side of the Sierras, down stream of historic hydraulic gold mining and processing; and South San Francisco Bay (CA), the estuarine component down stream of the New Almaden Mercury Mines. Average benthic flux of total-dissolved mercury was highest in Lahontan Reservoir ( ˜1400 pmol/m2/hr), followed by Camp Far West Reservoir ( ˜180 pmol/m2/hr), and lowest in South San Francisco Bay ( ˜50 pmol/m2/hr). In spite of this wide range of values, and the unique character of each watershed (e.g., forested vs. urbanized), all three systems exhibited quantitatively significant mercury benthic fluxes relative to riverine inputs. That is, areally averaged benthic fluxes (thus, expressed as annual loads) were of similar or greater magnitude relative to riverine loads. System-averaged values of dissolved methylmercury fluxes were similar for South San Francisco Bay (undetectable) and Camp Far West Reservoir (average of ˜0 pmol/m2/hr; some fluxes undetectable), and only slightly higher in Lahontan Reservoir ( ˜2 pmol/m2/hr). Similarly, system averaged potential rates of methylmercury production (by sulfate-reducing bacteria; as assessed by 203Hg(II) radiotracer studies) in the surficial sediment were not significantly different among the three locations. However, within-system variability was approximately an order of magnitude in each case

  15. Effect of DMPS and DMSA on the placental and fetal disposition of methylmercury.

    PubMed

    Bridges, C C; Joshee, L; Zalups, R K

    2009-09-01

    Methylmercury (CH3Hg+) is a serious environmental toxicant. Exposure to this metal during pregnancy can cause serious neurological and developmental defects in a developing fetus. Surprisingly, little is known about the mechanisms by which mercuric ions are transported across the placenta. Although it has been shown that 2,3-dimercaptopropane-1-sulfonate (DMPS) and 2,3-dimercaptosuccinic acid (DMSA) are capable of extracting mercuric ions from various organs and cells, there is no evidence that they are able to extract mercury from placental or fetal tissues following maternal exposure to CH3Hg+. Therefore, the purpose of the current study was to evaluate the ability of DMPS and DMSA to extract mercuric ions from placental and fetal tissues following maternal exposure to CH3Hg+. Pregnant Wistar rats were exposed to CH3HgCl, containing [203Hg], on day 11 or day 17 of pregnancy and treated 24 h later with saline, DMPS or DMSA. Maternal organs, fetuses, and placentas were harvested 48 h after exposure to CH3HgCl. The disposition of mercuric ions in maternal organs and tissues was similar to that reported previously by our laboratory. The disposition of mercuric ions in placentas and fetuses appeared to be dependent upon the gestational age of the fetus. The fetal and placental burden of mercury increased as fetal age increased and was reduced by DMPS and DMSA, with DMPS being more effective. The disposition of mercury was examined in liver, total renal mass, and brain of fetuses harvested on gestational day 19. On a per gram tissue basis, the greatest amount of mercury was detected in the total renal mass of the fetus, followed by brain and liver. DMPS and DMSA reduced the burden of mercury in liver and brain while only DMPS was effective in the total renal mass. The results of the current study are the first to show that DMPS and DMSA are capable of extracting mercuric ions, not only from maternal tissues, but also from placental and fetal tissues following maternal

  16. Transport of the glutathione-methylmercury complex across liver canalicular membranes on reduced glutathione carriers.

    PubMed

    Dutczak, W J; Ballatori, N

    1994-04-01

    Methylmercury transport across liver canalicular membranes into bile, a major route of excretion of this toxic compound, is dependent upon intracellular GSH, and a glutathione-methylmercury complex (CH3Hg.SG) has been detected in liver tissue and bile. To examine whether the CH3Hg.SG complex is itself transported across the canalicular membrane and to identify the transport system involved, studies were performed in isolated rat liver canalicular plasma membrane vesicles. Uptake of CH3(203)Hg.SG (10 microM) into an osmotically active space was temperature-sensitive and unaffected by either ATP (5 mM) or an inwardly directed Na+ gradient (100 mM); however, CH3Hg.SG uptake was enhanced by a valinomycin-induced K+ diffusion potential (inside-positive) indicating that its transport was electrogenic. Transport of CH3Hg.SG exhibited saturation kinetics with both high affinity (Km = 12 +/- 2 microM, Vmax = 0.23 +/- 0.02 nmol.mg-1.20 s-1) and low affinity (Km = 1.47 +/- 0.22 mM, Vmax = 1.23 +/- 0.14 nmol.mg-1.20 s-1) components. Uptake of this complex was inhibited by GSH, the GSH analog ophthalmic acid, S-methyl, S-ethyl, S-butyl, S-hexyl, S-octyl, and S-dinitrophenyl glutathione, but not by GSSG, bile acids, amino acids, and P-glycoprotein inhibitors. Furthermore, GSH competitively inhibited (Ki = 83 microM) and trans-stimulated CH3Hg.SG uptake into the canalicular vesicles. These studies provide the first kinetic characterization of a transport system for glutathione-mercaptides and indicate that CH3Hg.SG is not a substrate for the ATP-dependent, canalicular GSSG or glutathione S-conjugate carriers, but appears to be a substrate for canalicular carriers that also transport GSH. Because efflux systems for GSH are found in all mammalian cells, transport of glutathione-metal complexes by such carriers may be a common mechanism for the removal of methylmercury and possibly other metals from cells.

  17. Decrease in platelet activating factor stimulated phosphoinositide turnover during storage of human platelets in plasma

    SciTech Connect

    Carter, M.G.; Shukla, S.D. )

    1987-05-01

    Human platelet concentrate from the American Red Cross Blood Center was stored at 24{degree}C in a shaker and aliquots were taken out at time intervals aseptically. Platelet activating factor (PAF) stimulated turnover of phosphoinositide (PPI) was monitored by assaying {sup 32}P incorporation into phosphoinositides using platelet rich plasma (PRP). Platelets in PRP were incubated with 1 {times} 10{sup {minus}7} M PAF at 37{degree}C with gentle shaking and after 5 min their lipids were extracted and analysed by TLC for {sup 32}P-phosphoinositides. The percent stimulation of {sup 32}P incorporation by PAF (over control) into PPI was approximately 250, 100, 60, 25 and 20 on days 1, 2, 3, 5 and 6, respectively. This indicated a dramatic decrease in PAF responsive turnover of platelet PPI during storage. These findings have important implications in relation to PAF receptor activity and viability of platelets at different periods of storage.

  18. Phosphorylation of ribosomal proteins induced by auxins in maize embryonic tissues. [Zea mays

    SciTech Connect

    Perez, L.; Aguilar, R.; Mendez, A.P.; de Jimenez, E.S.

    1990-11-01

    The effect of auxin on ribosomal protein phosphorylation of germinating maize (Zea mays) tissues was investigated. Two-dimensional gel electrophoresis and autoradiography of ({sup 32}P) ribosomal protein patterns for natural and synthetic auxin-treated tissues were performed. Both the rate of {sup 32}P incorporation and the electrophoretic patterns were dependent on {sup 32}P pulse length, suggesting that active protein phosphorylation-dephosphorylation occurred in small and large subunit proteins, in control as well as in auxin-treated tissues. The effect of ribosomal protein phosphorylation on in vitro translation was tested. Measurements of poly(U) translation rates as a function of ribosome concentration provided apparent K{sub m} values significantly different for auxin-treated and nontreated tissues. These findings suggest that auxin might exert some kind of translational control by regulating the phosphorylated status of ribosomal proteins.

  19. Prolonged RNA changes in the Hermissenda eye induced by classical conditioning

    SciTech Connect

    Nelson, T.J.; Alkon, D.L. )

    1988-10-01

    The incorporation of {sup 32}P into mRNA and the total amount of mRNA were increased 3- to 4-fold in eyes isolated from Hermissenda crassicornis trained to associate light with rotation on a turntable compared with animals trained with equal numbers of light and rotation events presented randomly and with naive animals. Incorporation of {sup 32}P into poly(A){sup {minus}} RNA was reduced by as much as 60%. The RNA changes were strongly correlated with the degree of learning and could not be accounted for by changes in ({sup 32}P)ATP content. The RNA changes were maximal at 24 hr and were still detectable after 4 days, indicating that associative conditioning produces a period of increased DNA transcription that could be an intermediate step in memory consolidation. The RNA changes may in part account for recently observed conditioning-specific changes in the synthesis rates of specific proteins.

  20. Direct nonchromatographic assay for 1-acyl-sn-glycerol-3-phosphate acyltransferase

    SciTech Connect

    Rajasekharan, R.; Ray, T.K.; Cronan, J.E. Jr.

    1988-09-01

    1-Acyl-sn-glycerol-3-phosphate acyltransferase (also called lysophosphatidic acid acyltransferase) which catalyzes the acylation of 1-acyl-sn-glycerol-3-phosphate to phosphatidic acid is generally assayed by the use of a radioactive substrate followed by a time-consuming chromatographic separation of substrate and product. We report a direct and highly sensitive nonchromatographic assay for this enzyme based on the ability of Escherichia coli alkaline phosphatase to dephosphorylate 1-acyl-sn-glycerol-3-phosphate but not phosphatidic acid. This selective hydrolysis coupled with the use of /sup 32/P-labeled 1-acyl-sn-glycerol-3-phosphate as substrate permits measurement of the product, /sup 32/P-labeled phosphatidic acid by solvent extraction or precipitation. We also report a series of enzymatic reactions for the efficient conversion of /sup 32/Pi to /sup 32/P-labeled 1-acyl-sn-glycerol-3-phosphate.

  1. Further investigations into hormone-directed transport in stems.

    PubMed

    Bowen, M R; Wareing, P F

    1971-06-01

    The effect of IAA and three synthetic auxins on the upward movement of (32)P-orthophosphate and (14)C-sucrose has been investigated in decapitated stems of Phaseolus vulgaris L., Pisum sativum L., Coleus blumci L. and Helianthus annuus L. IAA greatly enhanced the accumulation of (32)P-orthophosphate in Phaseolus vulgaris and Pisum sativum, whereas in C. blumei and H. annuus it did not. 2,4-D like IAA, caused an increase in (14)C-sucrose and (32)P-orthophosphate accumulation in Phaseolus vulgaris but, unlike IAA, caused no increase in Pisum sativum. The downward transport of (14)C-IAA, (14)C-NAA, (14)C-2,4-D and (14)C-2,4,5-T from the decapitated apex was also studied. Results are discussed in terms of current theories of hormone-directed transport.

  2. Proteins that interact with GTP during sporulation of Bacillus subtilis

    SciTech Connect

    Mitchell, C.; Vary, J.C. )

    1989-06-01

    During sporulation of Bacillus subtilis, several proteins were shown to interact with GTP in specific ways. UV light was used to cross-link ({alpha}-{sup 32}P)GTP to proteins in cell extracts at different stages of growth. After electrophoresis, 11 bands of radioactivity were found in vegetative cells, 4 more appeared during sporulation, and only 9 remained in mature spores. Based on the labeling pattern with or without UV light to cross-link either ({alpha}-{sup 32}P)GTP or ({gamma}-{sup 32}P)GTP, 11 bands of radioactivity were apparent guanine nucleotide-binding proteins, and 5 bands appeared to be phosphorylated and/or guanylated. Similar results were found with Bacillus megaterium. Assuming the GTP might be a type of signal for sporulation, it could interact with and regulate proteins by at least three mechanisms.

  3. An enzymic radiochemical method for determining phosphatidylglycerol in amniotic fluid

    SciTech Connect

    Siegel, L.; Walker, S.I.; Robin, N.I.

    1983-05-01

    We describe an enzymic quantification of phosphatidylglycerol in amniotic fluid. Phosphatidylglycerol is hydrolyzed in alkali and the resulting glycerol is then enzymatically phosphorylated with adenosine 5'-(gamma-/sup 32/P)triphosphate to yield glycero(/sup 32/P)phosphate. After removal of excess (gamma-/sup 32/P)ATP by charcoal, the radioactivity of the glycerophosphate is measured in a liquid scintillation counter. Triglyceride in the amniotic fluid is hydrolyzed by lipase before extraction and thus does not interfere with the analysis. This method is specific for phosphatidylglycerol. Preliminary studies suggest that a phosphatidylglycerol value greater than or equal to 10 nmol/mL correlates with fetal lung maturity.

  4. Phosphorylation of Ribosomal Proteins Induced by Auxins in Maize Embryonic Tissues

    PubMed Central

    Pérez, Laura; Aguilar, Raúl; Méndez, Alma Pérez; de Jiménez, Estela Sánchez

    1990-01-01

    The effect of auxin on ribosomal protein phosphorylation of germinating maize (Zea mays) tissues was investigated. Two-dimensional gel electrophoresis and autoradiography of [32P] ribosomal protein patterns for natural and synthetic auxin-treated tissues were performed. Both the rate of 32P incorporation and the electrophoretic patterns were dependent on 32P pulse length, suggesting that active protein phosphorylation-dephosphorylation occurred in small and large subunit proteins, in control as well as in auxin-treated tissues. The effect of ribosomal protein phosphorylation on in vitro translation was tested. Measurements of poly(U) translation rates as a function of ribosome concentration provided apparent Km values significantly different for auxin-treated and nontreated tissues. These findings suggest that auxin might exert some kind of translational control by regulating the phosphorylated status of ribosomal proteins. Images Figure 3 PMID:16667828

  5. Studies on the Biochemistry and Fine Structure of Silica Shell Formation in Diatoms. Photosynthesis and Respiration in Silicon-Starvation Synchrony of Navicula pelliculosa 1

    PubMed Central

    Coombs, J.; Spanis, C.; Volcani, B. E.

    1967-01-01

    Rates of photosynthesis, measured by oxygen electrode or by 14CO2 fixation, dark respiration and 32P-phosphate incorporation are reported for the silicon-starvation synchrony of the fresh water diatom Navicula pelliculosa. During late exponential growth the rates were consistent with increase in carbon mass. During silicon starvation, rates of carbon dioxide fixation, oxygen evolution and 32P incorporation fell, and the saturating light intensity decreased from 27,000 lux to 5000 lux. Reintroduction of silicon led to immediate transients in all parameters studied, followed by a prolonged increase in rate of dark respiration and a gradual increase in apparent photosynthesis. During release of daughter cells, the rates of dark respiration decreased as photosynthesis and 32P incorporation increased. These results are discussed in relation to effects of silicon on the energy metabolism of the diatom. PMID:6080873

  6. Bioassay for neutron-dose estimations of three patients in the JCO criticality accident in Tokai-mura by measuring beta-ray emitters.

    PubMed

    Takeda, H; Miyamoto, K; Yukawa, M; Nishimura, Y; Watanabe, Y; Kim, H S; Fuma, S; Kuroda, N; Kouno, F; Joshima, H; Hirama, T; Akashi, M

    2001-09-01

    The measurement of beta-emitters in biological samples (hair, urine and bone) from three patients in the JCO criticality accident was performed to assess the neutron dose to individuals. The result of the measurements of 32P in hair and urine collected immediately after the accident showed that sufficient 32P activities had been induced in the hair by fast neutrons and in the urine by thermal neutrons to know the severity of the exposure to the individuals and to the position. From the measurement of 32P and 45Ca in bone from various anatomical parts of two patients who died 82 and 210 days after the accident, it was suggested that the distribution of the induced beta-emitters activities could prove the position and posture of the patients at the moment of exposure.

  7. In-vivo effect of Lithospermum ruderale on LHRH activity in the chick

    SciTech Connect

    Zeller, F.J.; Breneman, W.R.

    1981-07-01

    Lithospermum (LSPM) plant extracts can inhibit gonadotropin action in mammals and birds. The experiments reported in this paper were performed to note the possible effect of LSPM on the ability of LHRH to stimulate /sup 32/P testis uptake in immature chicks. LHRH injected 2 h before autopsy significantly increased /sup 32/P uptake in these animals. Water extracts of Lithospermum ruderale roots were administered subcutaneously at various concentrations and times before LHRH. LSPM injections of 2.0, 4.0, or 8.0 mg equivalents of dried material given 10 h before LHRH significantly suppressed the releasing hormone effect. 4.0 mg LSPM was not inhibitory at 16, 22, 38 or 46 h before LHRH. Interestingly, LHRH had a greater effect, as measured by /sup 32/P testis uptake, when given to these latter 3 groups then when given to controls. This suggests that LSPM may have prevented the release but not the synthesis of anterior pituitary gonadotropins.

  8. Phosphorus Compounds in Translocating Phloem

    PubMed Central

    Bieleski, R. L.

    1969-01-01

    Phosphate-32P was introduced into a turnip leaf, and 3 hr later, the vascular bundles were stripped from the petiole and their phosphate ester pattern was studied. The pattern did not alter along their length and was like that of other tissues. Pumpkin leaves were painted with phosphate-32P; and later, the petioles were cut, the sieve tube exudates were collected and their phosphate ester patterns were studied. Exudates collected after 10 min had a high proportion of their 32P present in Pi and nucleoside triphosphates, while exudates collected after long translocation times (4-22 hr) had a lower proportion in these, and a higher proportion in hexose monophosphates and UDP glucose. In general, the ester patterns were like those of other tissues. The results indicate that sieve tubes are metabolically active, and that Pi is the primary form in which phosphorus moves in the phloem. Images PMID:16657091

  9. Enzymatic method for radiolabelling vertebrate vitellogenin

    SciTech Connect

    Opresko, L.; Wiley, H.S.

    1984-08-01

    Phosphoprotein kinases from Xenopus and chicken liver have been purified and these enzymes have been used to label Xenopus vitellogenin, a phosphoprotein, to high specific activity with (..gamma..-/sup 32/P)ATP. The enzymes were isolated by (NH/sub 4/)/sub 2/SO/sub 4/ fractionation followed by chromatography on DE-52 cellulose and phosphocellulose. This procedure resulted in greater than 20,000-fold enrichment for the enzymes. Both enzyme preparations were used to selectively label vitellogenin in the serum of estrogen-treated animals. Thus, isolation of the vitellogenin prior to radiolabeling was not necessary. The (/sup 32/P)vitellogenin labeled in situ was incorporated by oocytes at a rate similar to (/sup 32/P)vitellogenin labeled in vivo, was translocated to the yolk platelets, and was correctly processed into the yolk proteins.

  10. Stimulatory effect of bombesin on phosphoinositide metabolism in the rat pineal gland.

    PubMed

    Novotná, R; Novotný, I

    1997-10-03

    The pineal gland is under complex peptidergic nervous control originating from hypothalamic nuclei. The daily rhythm of bombesin-like peptide in the hypothalamus suggests that this neuropeptide, similarly as other neuropeptides, might be involved in modulation of the physiological activity of the pineal gland. In our experiments we studied the mechanism of signal transduction of bombesin in the isolated pineal glands of rats. The phosphoinositide signalling system was examined by measuring 32P-labelling of phosphatidylinositol (PI), phosphatidylinositol phosphate (PIP) and phosphatidylinositol bisphosphate (PIP2), which reflects phosphoinositide cycle activation. Bombesin induced a significant increase in 32P-labelling of PI, PIP and PIP2. The antagonist of this neuropeptide, (D-Phe12-Leu14)-bombesin, suppressed the increase in 32P-labelling of all phosphoinositides. Bombesin was without effect on cAMP dependent protein phosphorylation. The data indicate that bombesin activates the PI signalling system via specific receptors.

  11. Superconductive ADC Project Fabrication Package. Final Design Review Package (Briefing Charts)

    DTIC Science & Technology

    2010-09-07

    Simulation Results Iin 1.45mV 2.5Ω 1pH 4pH 100fF 310uA 1fΩ Ic = 300uA Rn = 0.8Ω Cs = 0.32pF Ic = 300uA Rn = 0.8Ω Cs = 0.32pF Iin Vout Vout Ic = 500uA Rn...0.55Ω Cs = 0.32pF ONR Superconductive ADC CLIN/SLIN 0001AD September 2010, Brad Perranoski Pg. 17 Modulator Design Documentation Comparator Design...Comparator Design - Cadence Schematic & Simulation Comparator Testbench Simulation Results 1.45mV 2.5Ω 1pH 4pH 100fF 310uA 1fΩSine wave 100uApk Iin Ic

  12. Protein phosphorylation as a mechanism for regulation of spinach leaf sucrose-phosphate synthase activity

    SciTech Connect

    Huber, J.L.A.; Huber, S.C. )

    1989-04-01

    Protein phosphorylation has been identified as a mechanism for the light-dark regulation of spinach sucrose-phosphate synthase (SPS) activity, previously shown to involve some type of covalent modification of the enzyme. The 120 kD subunit of SPS in extracts of light-treated leaves was labeled with {sup 32}P in the presence of ({gamma}-{sup 32}P) ATP. In this in vitro system, {sup 32}P incorporation into light-activated SPS was dependent upon ATP and magnesium concentrations as well as time, and was closely paralleled by inactivation of the enzyme. The soluble protein kinase involved in the interconversion of SPS between activated and deactivated forms may be specific for SPS as it co-purifies with SPS during partial purification of the enzyme. The kinase appears not to be calcium activated and no evidence has been obtained for metabolite control of SPS phosphorylation/inactivation.

  13. Succinyl-CoA synthetase is a phosphate target for the activation of mitochondrial metabolism.

    PubMed

    Phillips, Darci; Aponte, Angel M; French, Stephanie A; Chess, David J; Balaban, Robert S

    2009-08-04

    Succinyl-CoA synthetase (SCS) is the only mitochondrial enzyme capable of ATP production via substrate level phosphorylation in the absence of oxygen, but it also plays a key role in the citric acid cycle, ketone metabolism, and heme synthesis. Inorganic phosphate (P(i)) is a signaling molecule capable of activating oxidative phosphorylation at several sites, including NADH generation and as a substrate for ATP formation. In this study, it was shown that P(i) binds the porcine heart SCS alpha-subunit (SCSalpha) in a noncovalent manner and enhances its enzymatic activity, thereby providing a new target for P(i) activation in mitochondria. Coupling 32P labeling of intact mitochondria with SDS gel electrophoresis revealed that 32P labeling of SCSalpha was enhanced in substrate-depleted mitochondria. Using mitochondrial extracts and purified bacterial SCS (BSCS), we showed that this enhanced 32P labeling resulted from a simple binding of 32P, not covalent protein phosphorylation. The ability of SCSalpha to retain its 32P throughout the SDS denaturing gel process was unique over the entire mitochondrial proteome. In vitro studies also revealed a P(i)-induced activation of SCS activity by more than 2-fold when mitochondrial extracts and purified BSCS were incubated with millimolar concentrations of P(i). Since the level of 32P binding to SCSalpha was increased in substrate-depleted mitochondria, where the matrix P(i) concentration is increased, we conclude that SCS activation by P(i) binding represents another mitochondrial target for the P(i)-induced activation of oxidative phosphorylation and anaerobic ATP production in energy-limited mitochondria.

  14. The Measurements of Affinity Constants of Osmptrophs For P and Release Rates of P From Microbial Food Web From Late Summer To Early Winter In The Bay of Villefranche, The NW Mediterranean

    NASA Astrophysics Data System (ADS)

    Tanaka, T.; Rassoulzadegan, F.; Thingstad, T. F.

    Orthophosphate uptake by bacteria and phytoplankton was measured with 32P for wa- ter from the Bay of Villefranche (NW Mediterranean) from September to December 2001. The 32P release rate was measured for waters with different size of micro- bial community. During this period, the thermocline was gradually declined and the vertical mixing started, showing the transition from nutrient depleted to replete pe- riods. Before the vertical mixing started, orthophosphate turnover time ranged from 1 to 5 h. The fraction of orthophosphate uptake was dominated (mean 70%) by the 0.6-2 µm fraction where cyanobacteria biomass was dominant. Based on the theo- retical considerations proposed by Thingstad and Rassoulzadegan (1999), the affinity constants were estimated for bacteria, cyanobacteria and autotrophic nanoflagellates, respectively. They ranged from 0.001 to 0.028 L/nM-P/h for bacteria, from 0.047 to 0.103 L/nM-P/h for cyanobacteria and from 0.002 to 0.032 L/nM-P/h for autotrophic nanoflagellates when 32P turnover time was relatively shorter (<5 h). This may sup- port the previous report in which cyanobacteria were dominant in the primary pro- duction in autumn at the study site. The 32P release rates after the addition of cold chase were always less than 0.5%/h. This release appeared to be from the 0.2-0.6 or 0.6-2 µm fractions. The increase of 32P in the >2 µm fraction suggests P transfer by predation. Virus appeared to not contribute to 32P release from bacteria during the study period. These results indicate long P turnover times within particulate fraction and thus less contribution of P release from microbial food web.

  15. Photoaffinity labeling of high affinity nicotinic acid adenine dinucleotide phosphate (NAADP)-binding proteins in sea urchin egg.

    PubMed

    Walseth, Timothy F; Lin-Moshier, Yaping; Jain, Pooja; Ruas, Margarida; Parrington, John; Galione, Antony; Marchant, Jonathan S; Slama, James T

    2012-01-20

    Nicotinic acid adenine dinucleotide phosphate (NAADP) is a messenger that regulates calcium release from intracellular acidic stores. Recent studies have identified two-pore channels (TPCs) as endolysosomal channels that are regulated by NAADP; however, the nature of the NAADP receptor binding site is unknown. To further study NAADP binding sites, we have synthesized and characterized [(32)P-5-azido]nicotinic acid adenine dinucleotide phosphate ([(32)P-5N(3)]NAADP) as a photoaffinity probe. Photolysis of sea urchin egg homogenates preincubated with [(32)P-5N(3)]NAADP resulted in specific labeling of 45-, 40-, and 30-kDa proteins, which was prevented by inclusion of nanomolar concentrations of unlabeled NAADP or 5N(3)-NAADP, but not by micromolar concentrations of structurally related nucleotides such as NAD, nicotinic acid adenine dinucleotide, nicotinamide mononucleotide, nicotinic acid, or nicotinamide. [(32)P-5N(3)]NAADP binding was saturable and displayed high affinity (K(d) ∼10 nM) in both binding and photolabeling experiments. [(32)P-5N(3)]NAADP photolabeling was irreversible in a high K(+) buffer, a hallmark feature of NAADP binding in the egg system. The proteins photolabeled by [(32)P-5N(3)]NAADP have molecular masses smaller than the sea urchin TPCs, and antibodies to TPCs do not detect any immunoreactivity that comigrates with either the 45-kDa or the 40-kDa photolabeled proteins. Interestingly, antibodies to TPC1 and TPC3 were able to immunoprecipitate a small fraction of the 45- and 40-kDa photolabeled proteins, suggesting that these proteins associate with TPCs. These data suggest that high affinity NAADP binding sites are distinct from TPCs.

  16. GTP binding to the. beta. -subunit of tubulin is greatly reduced in Alzheimers disease

    SciTech Connect

    Khatoon, S.; Slevin, J.T.; Haley, B.E.

    1987-05-01

    A decrease occurs (80-100%) in the (/sup 32/P)8N/sub 3/GTP photoinsertion into a cytosolic protein (55K M/sub r/) of Alzheimer's (AD) brain, tentatively identified as the ..beta..-subunit of tubulin (co-migration with purified tubulin, concentration dependence of interaction with GTP, ATP and their 8-azido photoprobes, and similar effects of Ca/sup 2 +/ and EDTA on photoinsertion). This agrees with prior observations of (/sup 32/P)8N/sub 3/GTP interactions with brain tubulin and a recent report on faulty microtubular assembly in AD brain. The decrease in (/sup 32/P)8N/sub 3/GTP photoinsertion into the 55K M/sub r/ protein of AD brain was in contrast with other photolabeled proteins, which remained at equal levels in AD and age-matched normal brain tissues. The 55K and 45K M/sub r/ were the two major (/sup 32/P)8N/sub 3/GTP photoinsertion species in non-AD brain. Of 5 AD brains, the photoinsertion of (/sup 32/P)8N/sub 3/GTP into the 55K M/sub r/ region was low or absent in 4 (55K/45K=0.1); one was 75% below normals (55K/45K=0.24). Total protein migrating at 55K M/sub r/ was similar in AD and controls. AD brain tubulin, while present, has its exchangeable GTP binding site on ..beta..-tubulin blocked/modified such that (/sup 32/P)8N/sub 3/GTP cannot interact normally with this site.

  17. GTP-blot analysis of small GTP-binding proteins. The C-terminus is involved in renaturation of blotted proteins.

    PubMed

    Klinz, F J

    1994-10-01

    Recombinant c-Ha-ras, ralA and rap2, but not rap1A or rap1B proteins retained their ability to bind [alpha-32P]GTP after SDS/PAGE and transfer to nitrocellulose. Recombinant c-Has-ras missing the C-terminal 23 amino acid residues failed to bind [alpha-32P]GTP after the blot, and the ability of recombinant ralA missing the C-terminal 28 amino acid residues to bind [alpha-32P]GTP was decreased many-fold. The presence of nonionic detergents of the polyoxyethylene type such as Tween 20, Triton X-100, Nonidet P40 or Lubrol PX in the incubation buffer was necessary to induce renaturation of blotted recombinant c-Ha-ras protein, whereas other types of detergents were ineffective. We propose that detergents of the polyoxyethylene type induce the refolding of some types of blotted small GTP-binding proteins and that the C-terminus is involved in the refolding process. Membranes from NIH3T3 fibroblasts overexpressing c-Ha-ras protein showed much weaker binding of [alpha-32P]GTP as expected from the level of ras immunoreactivity. Treatment of fibroblasts with lovastatin, an inhibitor of hydroxymethylglutaryl-coenzyme A reductase, caused the accumulation of the unfarnesylated form of c-Ha-ras in the cytosol. Examination of [alpha-32P]GTP-binding and immunoreactivity for cytosolic and membrane-bound c-Ha-ras revealed that binding of [alpha-32P]GTP to unprocessed c-Ha-ras was increased about threefold compared to the same amount of processed c-Ha-ras. Our results demonstrate that detection and quantification of small GTP-binding proteins in eukaryotic cells by GTP-blot analysis is hampered by the fact that these proteins differ strongly in their ability to renature after blotting to nitrocellulose.

  18. The diagnosis of the gastric cancer by radionuclides and their labels, using catheter-type semiconductor radiation detector (CASRAD).

    PubMed

    Sassa, R; Iio, M; Sugita, T

    1980-01-01

    New method of differential diagnosis of gastric cancer was reported. After intravenous 32P administration, lesion was counted by small catheter-type readiation detector system under gastrofiberscopic direct vision control. Optimum counting time was examined in man and it was found to be between 20 to 50 hours after intravenous administration of 32P. Under the currently available technology of miniature detector, still beta-emitting carcinopohilic agent was found to be the agent of choice to this end, since gamma-emitting carcinophilic agents can not provide data originated only from the gastric mucosa in question.

  19. Biodegradable radioactive implants for glaucoma filtering surgery produced by ion implantation

    NASA Astrophysics Data System (ADS)

    Assmann, W.; Schubert, M.; Held, A.; Pichler, A.; Chill, A.; Kiermaier, S.; Schlösser, K.; Busch, H.; Schenk, K.; Streufert, D.; Lanzl, I.

    2007-04-01

    A biodegradable, β-emitting implant has been developed and successfully tested which prevents fresh intraocular pressure increase after glaucoma filtering surgery. Ion implantation has been used to load the polymeric implants with the β-emitter 32P. The influence of ion implantation and gamma sterilisation on degradation and 32P-fixation behavior has been studied by ion beam and chemical analysis. Irradiation effects due to the applied ion fluence (1015 ions/cm2) and gamma dose (25 kGy) are found to be tolerable.

  20. Effects of calcium, strontium, and barium ions on phosphorylation of hippocampal proteins in vitro.

    PubMed

    Hoch, D B; Wilson, J E

    1984-01-01

    Calcium ion alone or in the presence of added calmodulin stimulated in vitro transfer of 32P from [gamma 32P]ATP into several proteins of mitochondrial and synaptosomal particulate fractions from rat brain. Strontium ion was capable of substituting for calcium ion in this stimulation, but barium ion lacked this capacity. These results bring into question the hypothesis that calcium-dependent protein phosphorylation of synaptic proteins is intrinsic to neurotransmitter release during neurotransmission, but they do not rule out that possibility.

  1. Dephosphorylation of phosphoproteins and synthetic phosphopeptides. Study of the specificity of the polycation-stimulated and MgATP-dependent phosphorylase phosphatases.

    PubMed

    Agostinis, P; Goris, J; Waelkens, E; Pinna, L A; Marchiori, F; Merlevede, W

    1987-01-25

    The substrate specificity of different forms of polycation-stimulated (PCSH, PCSL, and PCSC) phosphorylase phosphatases and of the catalytic subunit of the MgATP-dependent protein phosphatase from rabbit skeletal muscle was investigated. This was done, with phosphorylase a as the reference substrate, using the synthetic phosphopeptides patterned after the phosphorylated sites of pyruvate kinase (type L) (Arg2-Ala-Ser(32P)-Val-Ala (S2), and its Thr(32P) substitute (T4)), inhibitor-1 (Arg4-Pro-Thr(32P)-Pro-Ala (T5), Arg2-Pro-Thr(32P)-Pro-Ala (T1), and its Ser(32P) substitute (S1)), and some modified phosphopeptides (Arg2-Ala-Thr(32P)-Pro-Ala (T2) and Arg2-Pro-Thr(32P)-Val-Ala (T3)), all phosphorylated by cyclic AMP-dependent protein kinase. In addition, casein(Thr-32P), phosphorylated by casein kinase-2, was also tested. The PCS phosphatases show a striking preference for the T4 configuration, PCSC being the least efficient. The catalytic subunit of the MgATP-dependent phosphatase was almost completely inactive toward all these substrates. As shown for the PCSH phosphatase, and comparing with T4, the two proline residues flanking the Thr(P) in T1 and T5, just as in inhibitor-1, drastically imparied the dephosphorylation by lowering the Vmax and not by affecting the apparent Km. The C-terminal proline (as in T2) by itself represents a highly unfavorable factor in the dephosphorylation. The critical effect of the sequence X-Thr(P)-Pro or Pro-Thr(P)-Pro (T1, T2, T5, and inhibitor-1) can be overcome by manganese ions. The additional finding that this is not the case with the Pro-Ser(P)-Pro sequence (S1) suggests that the effect of Mn2+ is highly substrate specific. These observations show the considerable importance of the primary structure of the substrate in determining the specificity of the protein phosphatases.

  2. Erding AS, Munchen, Germany. Revised Uniform Summary of Surface Weather Observations (RUSSWO)

    DTIC Science & Technology

    1971-10-22

    Base Operatio N None 37 ft Feb 56 bldg. AN/ CHO - 2 Mar 56 t Located on top of Bldg 431. No chge None 47 ft Oct 57 USAF KTAC APII 0-19 (OL-I) COTINRM...29.0 3 #3 32P 33@8 34v4 3549* 36,4 37.6 37 38.42 1 aso 800 21.ZI3 OyU , 29,0 32p4 3111.9 f7 3178 385 3p 4 p 40p6 41,8 42 2 28 4 ;4j7 0 8000 22,2 2$#30,0

  3. Characteristics of the norepinephrine-stimulated phosphatidylinositol turnover in rat pineal cell dispersions

    SciTech Connect

    Hauser, G.; Smith, T.L.

    1981-10-01

    Dispersed rat pineal cells can be used for the study of the phosphatidylinositol effect. The response to ( - )-norepinephrine of the incorporation of 32Pi into phospholipids is linear with time and cell concentration, stereospecific, and mediated through alpha-1-adrenergic receptors. Na+ in the incubation medium is obligatory for labeling of phosphatidylinositol and phosphatidylcholine by 32P. In the absence of K+, incorporation of 32P is drastically lowered and no stimulation by norepinephrine occurs. Rb+ can replace K+. Omission of Ca2+ or substitution with Sr2+ preferentially lowers incorporation of radioactivity into phosphatidylcholine. Mg2+ is not required for basal or stimulated labeling.

  4. Method for the preparation of size marker for synthetic oligonucleotides

    SciTech Connect

    Jing, G.Z.; Liu, A.; Leung, W.C.

    1986-01-01

    Terminal deoxynucleotidyltransferase was used for the addition of (..cap alpha..-/sup 32/P)dCTP to the 3'-OH termini of oligo(dT)/sub 12-18/. A collection of oligonucleotides with chain lengths ranging continuously from 13-mer to over 100-mer was generated. The reaction mixture was then mixed with oligo(dT)/sub 12-18/ labeled with (..gamma..-/sup 32/P)ATP by T/sub 4/ polynucleotide kinase. A sequence ladder with the bottom base as 12-mer was then formed. These oligonucleotides served as size marker for the purification and identification of oligonucleotides on polyacrylamide gel.

  5. Axoplasmic transport of microtubule-associated proteins in the rat sciatic nerve

    SciTech Connect

    Takenaka, T.; Inomata, K.

    1981-09-01

    /sup 32/P-ATP was injected into the L5 dorsal root ganglion and axoplasmic transport of the phosphorylate MA proteins 2, microtubule-associated proteins 2, was observed. After the injection of /sup 32/P-ATP, the nerve was dissected out at prescribed time intervals and sliced into 5-mm pieces. Each segment was electrophoresed on an SDS-polyacrylamide slab gel and subjected to autoradiography. A protein of 310,000 dalton was transported at a velocity of 6.6-10.6 mm/day in the axon with the electrophoretic mobility identical to that of MA proteins 2, one of the key components associated with the microtubules.

  6. Phosphorylation of intact erythrocytes in human muscular dystrophy

    SciTech Connect

    Johnson, R.M.; Nigro, M.

    1986-04-01

    The uptake of exogenous /sup 32/Pi into the membrane proteins of intact erythrocytes was measured in 8 patients with Duchenne muscular dystrophy. No abnormalities were noted after autoradiographic analysis. This contrasts with earlier results obtained when isolated membranes were phosphorylated with gamma-(/sup 32/P)ATP, and suggests a possible reinterpretation of those experiments.

  7. PURIFICATION AND RECOVERY OF BULKY HYDROPHOBIC DNA ADDUCTS

    EPA Science Inventory

    For many years 32P postlabeling has detected DNA adducts at very low levels and yet has not been able to identify unknown adducts. Mass spectrometry offers substantially improved identification powers, albeit at some loss in detection limits. With this ultimate utilization of ma...

  8. Early effects of Escherichia coli endotoxin infusion on vasopressin-stimulated breakdown and metabolism of inositol lipids in rat hepatocytes

    SciTech Connect

    Rodriguez de Turco, E.B.; Spitzer, J.A.

    1988-08-30

    The turnover of vasopressin-stimulated 32P-phosphoinositides and 32P-phosphatidic acid and accumulation of (2-3H)-inositol phosphates were examined in hepatocytes from rats infused i.v. with saline and E. coli endotoxin for 3 hrs. Within 60s of VP stimulation the decrease in phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate labeling as well as the increased uptake of 32P into phosphatidic acid were similar in both groups. However, at a later time (300s) the 32P-phosphatidylinositol turnover was greatly decreased concomitantly with a higher labeling of phosphatidic acid. The accumulation of (2-3H)-inositol phosphates in ET-cells was significantly decreased both at 30s and 600s after VP addition. The distribution of (2-3H)-inositol labeling accumulated in the different inositol phosphate fractions over the first 30s of VP stimulation showed a tendency to lower accumulation of inositol trisphosphate, and a significantly lower accumulation of inositol bisphosphate simultaneously with a higher labeling of the inositol tetrakisphosphate fraction. These observations reflect an early effect of ET-infusion on VP-stimulated inositol lipid turnover and on the subsequent metabolism of the released inositol phosphates.

  9. Rabbit synoviocyte inositol phospholipid metabolism is stimulated by hydroxyapatite crystals

    SciTech Connect

    Rothenberg, R.J.; Cheung, H.

    1988-04-01

    Inhibition of prostaglandin E2 synthesis partially ameliorates some aspects of synovitis, but joint destruction still progresses. Other aspects of phospholipid metabolism may play a role in synovial tissue pathophysiology. Products of phosphatidylinositol metabolism can activate intracellular processes in response to extracellular stimuli. We asked whether this pathway is activated in synoviocytes in monolayer tissue culture by the addition of hydroxyapatite (HA) crystals in medium. These crystals are found in pathological human synovial fluid. These crystals are associated with the secretion of degradative enzymes and with a destructive arthritis in humans. Rabbit synoviocyte cultures, previously incubated with (3H)inositol to label inositol phospholipids, were stimulated with the addition of hydroxyapatite (180 micrograms/ml) to the cultures. There was enhanced intracellular accumulation of (3H)inositol monophosphate (30-100%) after 4 h. This indicated an increased phospholipase C activity. The radioactivity in (3H)inositol bis- and trisphosphates was too low to reliably measure. The use of (32P)Pi allowed detection of these compounds. In the presence of HA, incorporation of (32P)Pi into phosphatidylinositol, phosphatidylinositol monophosphate, and phosphatidylinositol bisphosphate was increased. In addition, cultures exposed to (32P)Pi during stimulation with HA had an increased content of (32P)inositol monophosphate, bisphosphate, and trisphosphate.

  10. Problem-Solving Test: Analysis of DNA Damage Recognizing Proteins in Yeast and Human Cells

    ERIC Educational Resources Information Center

    Szeberenyi, Jozsef

    2013-01-01

    The experiment described in this test was aimed at identifying DNA repair proteins in human and yeast cells. Terms to be familiar with before you start to solve the test: DNA repair, germline mutation, somatic mutation, inherited disease, cancer, restriction endonuclease, radioactive labeling, [alpha-[superscript 32]P]ATP, [gamma-[superscript…

  11. A complex network of interactions between mitotic kinases, phosphatases and ESCRT proteins regulates septation and membrane trafficking in S. pombe.

    PubMed

    Bhutta, Musab S; Roy, Brinta; Gould, Gwyn W; McInerny, Christopher J

    2014-01-01

    Cytokinesis and cell separation are critical events in the cell cycle. We show that Endosomal Sorting Complex Required for Transport (ESCRT) genes are required for cell separation in Schizosaccharomyces pombe. We identify genetic interactions between ESCRT proteins and polo and aurora kinases and Cdc14 phosphatase that manifest as impaired growth and exacerbated defects in septation, suggesting that the encoded proteins function together to control these processes. Furthermore, we observed defective endosomal sorting in mutants of plo1, ark1 and clp1, as has been reported for ESCRT mutants, consistent with a role for these kinases in the control of ESCRT function in membrane traffic. Multiple observations indicate functional interplay between polo and ESCRT components: firstly, two-hybrid in vivo interactions are reported between Plo1p and Sst4p, Vps28p, Vps25p, Vps20p and Vps32p; secondly, co-immunoprecipitation of human homologues of Vps20p, Vps32p, Vps24p and Vps2p by human Plk1; and thirdly, in vitro phosphorylation of budding yeast Vps32p and Vps20p by polo kinase. Two-hybrid analyses also identified interactions between Ark1p and Vps20p and Vps32p, and Clp1p and Vps28p. These experiments indicate a network of interactions between ESCRT proteins, plo1, ark1 and clp1 that coordinate membrane trafficking and cell separation in fission yeast.

  12. Davis Monthan AFB Tucson, Arizona. Revised Uniform Summary of Surface Weather Observations (RUSSWO)

    DTIC Science & Technology

    1976-07-01

    Wet Bulb |. Dewe Porn , -, I-. . .. - S.A- " - " - .- ,; -. - " - "-- - - - - ------------- - - -- - - - -~~DATA PRUCESSI1G BRASCH 3 U OA1 ETAC...of Houts witlh Tompetat.4 < Relln Hum 0OF o32P F *7a 3 0F .93 F Total 0< Wert BAI ..... __ S Dew Pointt DATA PRUCESSING BRANCH 3 U&IA ETAC

  13. Identification of Lotus rhizobia by direct DNA hybridization of crushed root nodules

    SciTech Connect

    Cooper, J.E.; Bjourson, A.J.; Thompson, J.K.

    1987-07-01

    Hybridization of crushed Lotus pedunculatus root nodules with /sup 32/P-labeled total genomic DNA probes was used to identify Rhizobium loti and Bradyrhizobium sp. (Lotus rhizobia). Probes always hybridized with homologous target DNA and frequency with DNAs of other strains from the same genus. Intergeneric hybridization did not occur. Results were comparable to those from colony hybridization.

  14. Using weighted trait indices to select the best performing broccoli hybrids in multi-site and multi-year trials

    USDA-ARS?s Scientific Manuscript database

    Understanding and implementing evaluation data from vegetable trials conducted across multiple years and environments by multiple raters presents numerous challenges. In order to select new broccoli hybrids suitable for eastern production, the SCRI East Coast Broccoli Project has conducted over 32 p...

  15. Modulation of phosphorylation of tocopherol and phosphatidylinositol by hTAP1/SEC14L2-mediated lipid exchange

    USDA-ARS?s Scientific Manuscript database

    The vitamin E derivative, alpha-tocopheryl phosphate (aTP), is detectable in cultured cells, plasma and tissues in small amounts, suggesting the existence of enzyme(s) with a-tocopherol (aT) kinase activity. Here, we characterize the production of aTP from aT and [g-32P]-ATP in primary human coronar...

  16. Effect of Phytase on Apparent Total Tract Digestibility of Phosphorus in Corn-Soybean Meal Diets Fed to 100 kg Pigs

    USDA-ARS?s Scientific Manuscript database

    Five experiments were conducted to investigate the ability of different sources of phytase supplemented to the diet at graded levels to improve apparent P digestibility in finishing pigs. A corn-soybean meal basal diet containing 0.50% Ca and 0.32% P (0.06% available P) was used in all experiments a...

  17. Underground resource allocation between individual networks of mycorrhizal fungi.

    PubMed

    Mikkelsen, Bolette L; Rosendahl, Søren; Jakobsen, Iver

    2008-01-01

    Fusions between individual mycelia of arbuscular mycorrhizal (AM) fungi have been observed in two-dimensional systems but never in soil systems. Here, phosphorus ((32)P) labelling was used to demonstrate nutrient transfer between individual mycelia and to investigate the possible role of anastomosis. Trifolium subterraneum colonized by Glomus mosseae were grown in root-retaining mesh bags, which were placed 20 cm apart. The mycelium of one plant, the donor, had access to (32)P-labelled soil placed adjacent to the mesh bag. Transfer of (32)P from the donor mycelium to the receiver plant was measured at three harvests. In a second-harvest control treatment the receiver was colonized by Glomus caledonium in order to determine whether transfer occurred by other means than hyphal fusions. Significant amounts of P were transferred to the receiver plant at the last harvests when the two mycelia of G. mosseae overlapped. The transfer probably occurred via anastomoses between the mycelia as no transfer of (32)P was detected between the mycelia of different fungi at the second harvest. The indicated ability of AM fungal mycelia to anastomose in soil has implications for the formation of large plant-interlinking functional networks, long-distance nutrient transport and retention of nutrients in readily plant-available pools.

  18. Problem-Solving Test: Analysis of DNA Damage Recognizing Proteins in Yeast and Human Cells

    ERIC Educational Resources Information Center

    Szeberenyi, Jozsef

    2013-01-01

    The experiment described in this test was aimed at identifying DNA repair proteins in human and yeast cells. Terms to be familiar with before you start to solve the test: DNA repair, germline mutation, somatic mutation, inherited disease, cancer, restriction endonuclease, radioactive labeling, [alpha-[superscript 32]P]ATP, [gamma-[superscript…

  19. Nucleotide sequence at the termini of the DNA of Bacillus subtilis phage phi 29.

    PubMed Central

    Escarmís, C; Salas, M

    1981-01-01

    Phage phi 29 DNA cannot be phosphorylated with polynucleotide kinase and [gamma-32P]ATP because of the presence of a viral protein covalently linked to the 5' termini. The 5' ends can, however, be made susceptible to phosphorylation by treatment with alkali and alkaline phosphatase. Restriction fragments Hpa II C and Hpa II F, corresponding to the right and left ends of phi 29 DNA, respectively, were labeled at the 5' ends with polynucleotide kinase and [gamma-32P]ATP or at the 3' ends with terminal transferase and [alpha-32P]ATP or [alpha-32P]cordycepin 5'-triphosphate. After a secondary cleavage of the labeled fragments, the sequence of the first 150-180 nucleotides at the termini of phi 29 DNA was determined by the method of Maxam and Gilbert. The ends of phi 29 DNA are flush, and a six-nucleotides-long inverted terminal repetition was found. The functional implications of the sequences determined are discussed. Images PMID:6262800

  20. IDENTIFICATION OF STEROCHEMICAL CONFIGERATION OF CYCLOPENTA[CD]PYRENE-DNA ADDUCTS IN STRAIN A/J MOUSE LUNG AND C3H10T1/2CL8

    EPA Science Inventory

    The definitive identification of stereochemical configurations of DNA adducts detected by 32P-postlabeling requires co-chromatography of adducts with synthetic chromatographic standards. Four major and several minor DNA adducts are formed by cyclopenta[cd]pyrene (CPP) in strain A...

  1. IDENTIFICATION OF STEROCHEMICAL CONFIGERATION OF CYCLOPENTA[CD]PYRENE-DNA ADDUCTS IN STRAIN A/J MOUSE LUNG AND C3H10T1/2CL8

    EPA Science Inventory

    The definitive identification of stereochemical configurations of DNA adducts detected by 32P-postlabeling requires co-chromatography of adducts with synthetic chromatographic standards. Four major and several minor DNA adducts are formed by cyclopenta[cd]pyrene (CPP) in strain A...

  2. G Protein Activation Stimulates Phospholipase D Signaling in Plants.

    PubMed Central

    Munnik, T.; Arisz, S. A.; De Vrije, T.; Musgrave, A.

    1995-01-01

    We provide direct evidence for phospholipase D (PLD) signaling in plants by showing that this enzyme is stimulated by the G protein activators mastoparan, ethanol, and cholera toxin. An in vivo assay for PLD activity in plant cells was developed based on the use of a "reporter alcohol" rather than water as a transphosphatidylation substrate. The product was a phosphatidyl alcohol, which, in contrast to the normal product phosphatidic acid, is a specific measure of PLD activity. When 32P-labeled cells were treated with 0.1% n-butanol, 32P-phosphatidyl butanol (32P-PtdBut) was formed in a time-dependent manner. In cells treated with any of the three G protein activators, the production of 32P-PtdBut was increased in a dose-dependent manner. The G protein involved was pertussis toxin insensitive. Ethanol could activate PLD but was itself consumed by PLD as transphosphatidylation substrate. In contrast, secondary alcohols (e.g., sec-butyl alcohol) activated PLD but did not function as substrate, whereas tertiary alcohols did neither. Although most of the experiments were performed with the green alga Chlamydomonas eugametos, the relevance for higher plants was demonstrated by showing that PLD in carnation petals could also be activated by mastoparan. The results indicate that PLD activation must be considered as a potential signal transduction mechanism in plants, just as in animals. PMID:12242371

  3. Ethanol stimulates calcium mobilization, phosphoinositide turnover and shape change in human platelets

    SciTech Connect

    Rubin, R.; Hoek, J.B.

    1987-05-01

    The effect of ethanol of cytosolic free calcium levels was investigated in human platelets which were loaded with the intracellular calcium indicator FURA-2. Administration of ethanol in the concentration range of 50-300 mM resulted in a transient, dose-dependent increase in cytosolic calcium, maximal within 5 seconds and returning to near basal levels over the next 5 minutes. Parallel incubations of platelets with the same concentrations of ethanol stimulated shape change as detected in an aggregometer. Chelation of external calcium with EGTA prior to the addition of ethanol reduced the calcium burst partially, but not completely, whereas shape change was largely unaffected. Addition of ethanol to /sup 32/P-labeled platelets resulted in a 16% decrease in the level of /sup 32/P-phosphatidylinositol 4,5-bisphosphate and a 14% increase in /sup 32/P-phosphatidylinositol 4-phosphate within 2 minutes. /sup 32/P-phosphatidic acid levels increased rapidly within 30 seconds and rose linearly thereafter. Phosphatidylinositol and phosphatidylcholine were unaffected by ethanol. The results indicate that ethanol mobilizes intracellular calcium by activation of phosphoinositide-specific phospholipase C, thereby stimulating platelet shape change.

  4. PHYTOTOXICITY

    EPA Science Inventory

    Handbook of Ecotoxicology. Second Edition.. Lewis Publishers, Boca Raton, FL. 32 p.

    Phytoplankton, benthic and epiphytic microalgae, and macroalgae are energy sources critical to most aquatic ecosystems. Changes in their density and composition can effect the chemical and...

  5. Effects of thyrotropin on the phosphorylation of histones and nonhistone phosphoproteins in micrococcal nuclease-sensitive and resistant thyroid chromatin

    SciTech Connect

    Cooper, E.; Spaulding, S.W.

    1983-05-01

    Actively transcribed regions of chromatin are more susceptible than bulk chromatin to digestion by nucleases, and useful information about the composition and structure of active chromatin may be obtained by studying the chromatin fragments released from nuclei by limited nuclease digestion. In the present study, we have used micrococcal nuclease to investigate the effects of TSH on protein phosphorylation in nuclease-sensitive fractions of calf thyroid chromatin. Batches of calf thyroid slices were incubated for 2 h with /sup 32/Pi, with or without 50 mU/ml TSH. Nuclei were then prepared and the distribution of /sup 32/P-labeled histones, high mobility group (HMG) proteins, and other acid-soluble phosphoproteins between micrococcal nuclease-sensitive and resistant fractions of chromatin was examined. TSH increased the amount of /sup 32/P incorporated into HMG 14 and the histones H1 and H3. Hormone-dependent increases in the /sup 32/P-labeling of H1 and H3 were not selectively associated with micrococcal nuclease-sensitive chromatin. In contrast, (/sup 32/P) HMG-14 was preferentially solubilized from nuclei by micrococcal nuclease. This lends support to the view that TSH-induced effects on the structure and function of transcriptionally active chromatin may be mediated in part by phosphorylation of HMG 14.

  6. Cyclic AMP-dependent phosphorylation of voltage-sensitive sodium channels in primary cultures of rat brain cells

    SciTech Connect

    Rossie, S.; Catterall, W.A.

    1986-03-05

    The ..cap alpha.. subunit of the voltage-sensitive Na channel from rat brain is phosphorylated by cAMP-dependent protein kinase in purified preparations and in synaptosomes. The authors have begun to study cAMP-dependent phosphorylation of Na channels in intact cells. Rat brain cells collected at embryonic day 15 and maintained in culture for approximately 21 days were subjected to treatments designed to increase intracellular cAMP. Cells were solubilized and Na channels were isolated by immunoprecipitation, then rephosphorylated with the catalytic subunit of cAMP-dependent protein kinase and /sup 32/P-ATP, to allow incorporation of /sup 32/P into available cAMP-dependent phosphorylation sites of Na channels. The amount of /sup 32/P incorporated into channel is inversely proportional to the extent of endogenous phosphorylation. Treatment of cells with forskolin inhibited rephosphorylation of Na channels, indicating that enhanced endogenous phosphorylation of channels had occurred. The effect of forskolin on cell surface Na channels occurred rapidly, was sustained over 30 min., and was half-maximal at 6..mu..M. 8-Br-cAMP, (EC/sub 50/-5mM) and isobutylmethylxanthine (EC/sub 50/-60..mu..M) also caused inhibition of /sup 32/P incorporation into Na channels. These results indicate that the extent of cAMP-dependent phosphorylation of voltage-sensitive Na channels in intact brain neurons is modified by changes in intracellular levels of cAMP.

  7. Insulin-like growth factor II receptor is phosphorylated by a tyrosine kinase in adipocyte plasma membranes

    SciTech Connect

    Corvera, S.; Whitehead, R.E.; Mottola, C.; Czech, M.P.

    1986-06-15

    Incorporation of /sup 32/P from (gamma-32P)ATP into tyrosine residues of the insulin-like growth factor (IGF)-II receptor was observed in a Triton X-100-insoluble fraction of rat adipocyte plasma membranes. IGF-II receptor phosphorylation proceeded to a stoichiometry of approximately 0.5 mol of phosphate/IGF-II binding site after 10 min of incubation at 4 degrees C. A Km for ATP of 6 microM was calculated for this phosphorylation reaction. Addition of IGF-II caused an approximately 2-fold increase in tyrosine phosphorylation of the IGF-II receptor in this preparation. In contrast, phosphorylation of angiotensin II by the Triton X-100 washed membranes was not stimulated by IGF-II. Incubation of purified receptor immobilized on IGF-II agarose or of receptor-enriched low density microsomal membranes with (gamma-32P)ATP did not result in appreciable incorporation of (/sup 32/P)phosphate into the IGF-II receptor nor into exogenous substrates. These data suggest that the IGF-II receptor is not a tyrosine protein kinase capable of autophosphorylation but that it is a substrate for a tyrosine protein kinase endogenous to the adipocyte plasma membrane. The stimulatory effect of IGF-II on the tyrosine phosphorylation of its receptor may be due to a conformational change which converts the receptor to a better substrate for this tyrosine kinase.

  8. Is there a relationship between phosphatidylinositol trisphosphate and F-actin polymerization in human neutrophils

    SciTech Connect

    Eberle, M.; Traynor-Kaplan, A.E.; Sklar, L.A.; Norgauer, J. )

    1990-10-05

    Stimulation of human neutrophils with the chemoattractant N-formyl peptide caused rapid polymerization of F-actin as detected by right angle light scatter and 7-nitrobenz-2-oxa-1,3-diazol (NBD)-phallacidin staining of F-actin. After labeling neutrophils with 32P, exposure to N-formyl peptide induced a fast decrease of phosphatidylinositol 4-bisphosphate (PIP)2, a slow increase of phosphatidic acid, and a rapid rise of phosphatidylinositol 4-trisphosphate (PIP3). Formation of PIP3 as well as actin polymerization was near maximal at 10 s after stimulation. Half-maximal response and PIP3 formation at early time points resulted from stimulation of neutrophils with 0.01 nM N-formyl peptide or occupation of about 200 receptors. Sustained elevation of PIP3, prolonged right angle light scatter response, and F-actin formation required higher concentrations of N-formyl peptide, occupation of thousands of receptors, and high binding rates. When ligand binding was interrupted with an antagonist, F-actin rapidly depolymerized, transient light scatter response recovered immediately, and elevated (32P)PIP3 levels decayed toward initial values. However, recovery of (32P)PIP2 was not influenced by the antagonist. Based on the parallel time courses and dose response of (32P) PIP3, the right angle light scatter response, and F-actin polymerization, PIP3 is more likely than PIP2 to be involved in modulation of actin polymerization and depolymerization in vivo.

  9. Effects of anti-C23 (nucleolin) antibody on transcription of ribosomal DNA in Chironomus salivary gland cells

    SciTech Connect

    Egyhazi, E.; Pigon, A. ); Chang, Jinhong; Ghaffari, S.H.; Dreesen, T.D.; Wellman, S.E.; Case, S.T.; Olson, M.O.J. )

    1988-10-01

    Protein C23 (also called nucleolin or 100-kDa nucleolar protein) is a major nucleolar phosphoprotein involved in ribosome biogenesis. To determine the effects of protein C23 on preribosomal RNA (pre-rRNA) synthesis anti-C23 antiserum was microinjected into nuclei of Chironomus tentans salivary glands. Transcription was measured by incubation of the glands with {sup 32}P-labeled RNA precursors followed by microdissection of nucleoli, RNA extraction, and electrophoretic analyses. Injection of the anti-C23 antibody caused a 2- to 3.5-fold stimulation of {sup 32}P incorporation into 38 S pre-rRNA. No stimulation was observed in salivary glands injected with preimmune serum or antiserum preabsorbed with protein C23. The stimulatory effect was selective for pre-rRNA as indicated by the lack of stimulation of {sup 32}P incorporation into extranucleolar RNA. Injection of the antiserum produced little or no effect on pre-RNA processing as measured by the relative amounts of {sup 32}P-labeled intermediate cleavage products of pre-rRNA in stimulated versus control glands. These results suggest that protein C23 not only is involved in ribosome assembly but also plays a role in regulating the transcription of the preribosomal RNA.

  10. Phosphate uptake in Saccharomyces cerevisiae Hansen wild type and phenotypes exposed to space flight irradiation.

    PubMed Central

    Berry, D; Volz, P A

    1979-01-01

    Rates of phosphate uptake were approximately twice as great for Saccharomyces cerevisiae single-cell phenotypic isolates exposed to space parameters as for the wild-type ground control. Quantitative determination of 32P was performed by liquid scintillation spectrometry utilizing Cerenkov radiation counting techniques. PMID:395899

  11. AERIAL OF SHUTTLE LANDING FACILITY [SLF] POURING CONCRETE ON RUNWAY

    NASA Technical Reports Server (NTRS)

    1975-01-01

    AERIAL OF SHUTTLE LANDING FACILITY [SLF] POURING CONCRETE ON RUNWAY KSC-375C-10036.32 108-KSC-375C-10036.32, P-21425, ARCHIVE-04501 Aerial oblique of Shuttle Landing Facility. Pouring concrete on runway. Direction North - Altitude 100'.

  12. Increased in vitro phosphorylation of rat liver nucleolar proteins following triiodothyronine administration.

    PubMed

    Fugassa, E; Gallo, G; Pertica, M

    1976-11-15

    It has been shown that triiodothyronine (Ta) administration to thyroidectomized rats induces an increase in the in vitro net 32P uptake into liver nucleolar proteins. Such an increase depends on a stimulation of the nucleolus-associated protein kinase activity and not on a lower dephosphorylation rate.

  13. Tumor-promoting phorbol diesters cause the phosphorylation of epidermal growth factor receptors in normal human fibroblasts at threonine-654.

    PubMed Central

    Davis, R J; Czech, M P

    1985-01-01

    The effect of tumor-promoting phorbol diesters to potentiate the action of epidermal growth factor (EGF) on cell proliferation is associated with phosphorylation of EGF receptors, acute depression of EGF binding, and inhibition of EGF receptor tyrosine kinase activity. In the present studies, normal human fibroblasts and A431 carcinoma cells were labeled with [32P]phosphate and treated with and without 10 nM 4 beta-phorbol 12 beta-myristate 13 alpha-acetate (PMA). The EGF receptors then were isolated by immunoprecipitation and digested with trypsin. Analysis of the labeled receptor phosphopeptides by reversed-phase HPLC revealed that PMA induces the phosphorylation of a unique phosphopeptide containing [32P]phosphothreonine. Comparison of several chemical and physical properties of the 32P-labeled phosphopeptide with the primary structure of the EGF receptor suggested the identify Lys-Arg-Thr(P)-Leu-Arg. This was confirmed by direct demonstration that a synthetic peptide of this structure comigrates during HPLC and electrophoresis with the 32P-labeled phosphopeptide isolated from the EGF receptors of normal human fibroblasts. The phosphorylated site on the peptide corresponds to threonine-654 of the EGF receptor, which is located on the cytoplasmic side of the plasma membrane nine residues distant from the transmembrane domain. These data indicate that phosphorylation of the EGF receptor in human fibroblasts and A431 cells at threonine-654 may regulate the EGF receptor tyrosine kinase activity and the binding of EGF. Images PMID:2984676

  14. PHYTOTOXICITY

    EPA Science Inventory

    Handbook of Ecotoxicology. Second Edition.. Lewis Publishers, Boca Raton, FL. 32 p.

    Phytoplankton, benthic and epiphytic microalgae, and macroalgae are energy sources critical to most aquatic ecosystems. Changes in their density and composition can effect the chemical and...

  15. Resolution of phosphoglucomatase and the 62-kDA acceptor for the glucosylphosphotransferase

    SciTech Connect

    Marchase, R.B.; Richardson, K.L.; Srisomsap, C.; Drake, R.R.; Haley, B.E. )

    1990-07-01

    The radioactive, photoactivatable labeling probe (beta-32P)5-azidouridine 5'-diphosphoglucose has recently been shown to label a 62-kDa protein in crude homogenates and in partially purified enzyme preparations without photoactivation. Here, we report that a portion of this radioactivity is due to labeling of phosphoglucomutase by contaminating levels of (32P)alpha Glc-1-P initially present at less than 1% of the total 32P. This conclusion is based in part on the ability of excess unlabeled alpha Glc-1-P and Glc-6-P, but not UDP-Glc, to block the labeling. In addition, the labeled protein in liver homogenates had a tryptic peptide pattern similar to that of authentic phosphoglucomutase. These findings, however, raised a second question. Assays for the UDP-Glc: glycoprotein glucosyl phosphotransferase (Glc phosphotransferase) have utilized (beta-32P)UDP-Glc and have resulted in the labeling of a small number of acceptors, including one of approximately 62 kDa. Despite the fact that these assays had routinely been performed in the presence of 1 mM alpha Glc-1-P, the coincidence in molecular weights led to these further studies. We conclude that the acceptor of approximately 62 kDa is distinct from phosphoglucomutase. This conclusion is based on differences in the time courses of incorporation, the specificity of blocking agents, the presence of covalently linked glucose, the products of acid hydrolysis and of beta-elimination, and isoelectric points.

  16. Metal: ATP characteristics of insulin- and epidermal growth factor-stimulated phosphorylation in detergent extracts of rat liver plasma membranes.

    PubMed

    Uhing, R J; Exton, J H

    1986-09-01

    The metal: ATP characteristics of insulin- and epidermal growth factor-(EGF)-stimulated protein kinase activities were examined in Nonidet P40 extracts of rat liver plasma membranes. The two kinase activities were capable of utilizing either manganese or magnesium, although differences were observed. Insulin-stimulated 32P incorporation into an Mr 95 000 protein exhibited a higher affinity for ATP in the presence of manganese compared to magnesium. At 200 microM ATP, insulin stimulated 32P incorporation into the Mr 95 000 protein 3- to 5-fold after 5 min in the presence of either metal. At 1 mM ATP, insulin-stimulated 32P incorporation was significantly greater in the presence of magnesium. In contrast, EGF-stimulated 32P incorporation into an Mr 170 000 protein exhibited similar ATP dependencies in the presence of magnesium or manganese. Basal phosphorylation of the Mr 170 000 protein was 2- to 3-fold higher in the presence of manganese, however. Since the higher basal phosphorylation persisted after chromatography on wheat germ lectin-Sepharose, it may represent an inherent activity of the receptor kinase. In the presence of magnesium: ATP, low concentrations of manganese enhanced both insulin- and EGF-stimulated phosphorylation of angiotensin II suggesting involvement of a second metal binding site which regulates the kinase activity. The results presented show major differences in the metal: ATP properties of the two major hormonally regulated protein kinase activities observed in detergent-extracted liver membranes.

  17. Selective Gene Regulation by Androgen Receptor in Prostate Cancer

    DTIC Science & Technology

    2013-10-01

    androgen receptor pathway in prostate cancer. Curr Opin Pharmacol, 2008. 8(4): p. 440-8. 6. Claessens, F., P. Alen , A. Devos, B. Peeters, G...Chem, 1996. 271(32): p. 19013-6. 7. Schoenmakers, E., P. Alen , G. Verrijdt, B. Peeters, G. Verhoeven, W. Rombauts, and F. Claessens, Differential DNA

  18. Detection of Listeria monocytogenes by using the polymerase chain reaction

    SciTech Connect

    Bessesen, M.T.; Luo, Q.; Blaser, M.J.; Ellison, R.T. III.; Rotbart. H.A. )

    1990-09-01

    A method was developed for detection of Listeria monocytogens by polymerase chain reaction amplification followed by agarose gel electrophoresis or dot blot analysis with {sup 32}P-labeled internal probe. The technique identified 95 of 95 L. monocytogenes strains, 0 of 12 Listeria strains of other species, and 0 of 12 non-Listeria strains.

  19. The Anopheles punctulatus complex: DNA probes for identifying the Australian species using isotopic, chromogenic, and chemiluminescence detection systems

    SciTech Connect

    Cooper, L.; Cooper, R.D.; Burkot, T.R. )

    1991-07-01

    Isotopic and enzyme-labeled species-specific DNA probes were made for the three known members of the Anopheles punctulatus complex of mosquitoes in Australia (Anopheles farauti Nos. 1, 2, and 3). Species-specific probes were selected by screening total genomic libraries made from the DNA of individual species with 32P-labeled DNA of homologous and heterologous mosquito species. The 32P-labeled probes for A. farauti Nos. 1 and 2 can detect less than 0.2 ng of DNA while the 32P-labeled probe for A. farauti No. 3 has a sensitivity of 1.25 ng of DNA. Probes were then enzyme labeled for chromogenic and chemiluminescence detection and compared to isotopic detection using 32P-labeled probes. Sequences of the probe repeat regions are presented. Species identifications can be made from dot blots or squashes of freshly killed mosquitoes or mosquitoes stored frozen, dried, and held at room temperature or fixed in isopropanol or ethanol with isotopic, chromogenic, or chemiluminescence detection systems. The use of nonisotopic detection systems will enable laboratories with minimal facilities to identify important regional vectors.

  20. Influence of nucleic acids and polysaccharides on phosphotransferase activity of preparations of secretory immunoglobulin A from human milk.

    PubMed

    Kit, Y Y; Semenov, D V; Kuligina, E V; Richter, V A

    2000-02-01

    The influence of nucleic acids (DNA, tRNA), synthetic oligonucleotides, and polysaccharides (lipopolysaccharides from Escherichia coli, heparin) on protein kinase and lipid kinase activities of preparations of human secretory immunoglobulin A (sIgA) has been studied. The preparations of sIgA were isolated from human milk by chromatography on the column with Protein A-Sepharose and DEAE-sorbent (sIgA1), by affinity chromatography of sIgA1 on DNA-cellulose (sIgA2), and by gel-filtration of sIgA1 in buffer containing 5% dioxane (sIgA3). Two 32P-labeled products with high and low electrophoretic mobility in polyacrylamide gel containing SDS were found after incubation of sIgA1 and sIgA2 with [gamma-32P]ATP. The product with low electrophoretic mobility was degraded in 10% trichloroacetic acid giving a radioactive background in lanes of the polyacrylamide gel. 32P-Labeled phospholipids were found among the phosphorylation products. Soluble and immobilized DNA increase lipid kinase activity of preparations of sIgA. In this case the secretory component and H-chains of sIgA were degraded. Fractions possessing lipid kinase activity were precipitated in the presence of heparin (1 mg/ml), and lipid kinase activity was separated from sIgA by gel-filtration in buffer containing 5% dioxane. 32P-Labeled products were formed in the presence of [gamma-32P]ATP as well as [32P]ortho-phosphoric acid. The influence of heparin and synthetic deoxy- and ribooligonucleotides on casein kinase activity of sIgA3 was studied. It was observed that deoxyribooligonucleotides in micromolar concentrations increased the rate of casein phosphorylation in the presence of sIgA3 and [gamma-32P]ATP. It has been proposed that catalytically active sIgA have an affinity to DNA (anti-DNA sIgA) and can be present in human milk as a part of lipoprotein complexes.

  1. Further studies on phosphorylated pituitary somatotropin (growth hormone)

    SciTech Connect

    Kornberg, L.J.; Liberti, J.P.

    1987-05-01

    This laboratory made the original observation that naturally-occurring ovine growth hormone (GH) is phosphorylated and that slices of pituitary glands from male rats synthesize and secrete /sup 32/P-GH. This observation has been extended to explore the generality of this process. After incubation in PO/sub 4/-free Ham's F-10 medium (PFH) or in saline/Hepes (SH) containing 300..mu..Ci /sup 32/Pi/mL, tissue and medium were separated and a cell extract was prepared. GH in the medium and extract was recovered by immunoprecipitation using rat GH antiserum. The samples were electrophoresed under denaturating conditions and processed for autoradiography. /sup 32/P-GH was characterized by the presence of a protein-staining band and radioactive area which migrated the same as authentic GH and /sup 125/I-GH. Slices of glands from male rats incubated for 2h in PFH secreted /sup 32/P-GH. Similar results were found upon incubation of slices from female rats in the presence of SH. Short-term incubations of acutely dispersed pituitary cells obtained from young and old male rats also synthesized and secreted /sup 32/P-GH. Thus, the production of /sup 32/P-GH occurs (a) in simple and complex incubaton media, (b) in slices and cells from glands from older and younger rats and (c) in female as well as male rats. Therefore, phosphorylation of GH appears to be a general phenomenon. The physiological action(s) of phosphorylated GH in growth and development is under study.

  2. Two distinct pools of membrane phosphatidylglycerol in Bacillus megaterium

    SciTech Connect

    Lombardi, F.J.; Fulco, A.J.

    1980-02-01

    The predominant membrane lipid in Bacillus megaterium ATCC 14581, phosphatidylglycerol (PG), is present in two distinct pools, as shown by (/sup 32/P)phosphate incorporation and chase experiments. One pool (PG/sub t/) undergoes rapid turnover of the phosphate moiety, whereas the second pool (PG/sub s/) exhibits metabolic stability in this group. The phosphate moiety of the other major phospholipid, phosphatidylethanolamine, is stable to turnover. (/sup 32/P)phosphate- and (2-/sup 3/H)glycerol-equilibrated cultures yielded the following glycerolipid composition: 56 mol% PG (34 mol% PG/sub t/ and 22 mol% PG/sub s/), 21 mol% phosphatidylethanolamine, 1 to 2 mol% phosphatidylserine, 20 mol% diglycerides, less than 0.5 mol% cardiolipin, and 0.2 to 0.4 mol% lysophosphatidylglycerol. Accumulation of PG was halted immediately after the addition of cerulenin, an inhibitor of de novo fatty acid synthesis, whereas phosphatidylethanolamine accumulation continued at the expense of the diglyceride and PG pools. Strikingly, initial rates of (/sup 32/P)phosphate incorporation into PG were unaffected by cerulenin. In control cultures at 35/sup 0/C, incorporation of (/sup 32/P)phosphate into PG exhibited a biphasic time course, whereas incorporation into phosphatidylethanolamine was concave upward and lagged behind that of PG during the initial rapid phase of PG incorporation. Finally, levels of lysophosphatidylglycerolexpanded rapidly after cerulenin addition at 20/sup 0/C, but not at 35/sup 0/C. Moreover, incorporation of (/sup 32/P)phosphate into lysophosphatidylglycerol lagged behind incorporation into PG in both the presence and absence of cerulenin at 20 and 35/sup 0/C.

  3. Magnesium-dependent adenosine triphosphatase as a marker enzyme for the plasma membrane of human polymorphonuclear leukocytes.

    PubMed

    Harlan, J; DeChatelet, L R; Iverson, D B; McCall, C E

    1977-02-01

    The adenosine triphosphatase (ATPase) activities of human polymorphonuclear leukocytes (PMNL) were studied with an assay that monitored the release of 32P-labeled inorganic pyrophosphate (32P1) from gamma-[32P]adenosine 5'-triphosphate (ATP). In cell homogenates, (Na+ + K+)-sensitive, ouabain-inhibitable ATPase comprised an insignificant fraction of the total ATPase activity. Additions of p-nitrophenyl phosphate and beta-glycerophosphate (substrates for nonspecific acid and alkaline phosphatases) and of tartrate (inhibitor of acid phosphatase) gave no indication of inhibition. This suggested that the assay was relatively specific for ATP hydrolysis. The activity was found to have a pH optimum of 8.7 and a Km for ATP of 0.6 mM. There was an absolute requirement for Mg2+, with other divalent cations substituting less efficiently. When the Mg2+-dependent ATPase activity of intact cells was compared with that in homogenized cells, no significant difference was observed. The activity in intact cells was linear with respect to incubation time up to at least l0 min. Trypan blue staining and lactate dehydrogenase assays revealed that greater than 92% of the PMNL remained intact and viable during the assay. No soluble ATPase was released from the cells under assay conditions. In following the distribution of gamma[32P]ATP and 32P2 counts became cell associated. Since the experimental evidence supports the observation that PMNL remain intact and viable and that ATP does not penetrate the cell under assay conditions, it is proposed that greater than 90% of the Mg2+-dependent ATPase of the human PMNL is associated with a plasma membrnae enzyme. This would qualify the enzyme for the role of a plasma membrane marker for future fractionation and isolation attempts.

  4. Altered turnover of polyphosphoinositides in the erythrocyte membrane of the spontaneously hypertensive rat

    SciTech Connect

    Koutouzov, S.; Marche, P.; Girard, A.; Meyer, P.

    1983-07-01

    The metabolism of inositol phospholipids of the erythrocyte membrane was compared in normotensive Wistar-Kyoto (WKY), spontaneously hypertensive (SHR), and stroke-prone SHR (SHR-SP) rats. This was performed on isolated ghost membranes by measuring the incorporation of /sup 32/P from ( gamma-/sup 32/P ) adenosine triphosphate (ATP) into the diphosphoinositides (DPI) and the triphosphoinositides (TPI) which were the only /sup 32/P-labeled phospholipids. /sup 32/P-labeling of TPI was altered in adult and 3-week-old SHR as well as in SHR-SP compared to WKY controls; the radioactivity associated with TPI in hypertensive rats was about 30% lower than that associated with TPI in age-matched normotensive controls. By contrast, the radioactivity associated with DPI was similar in both hypertensive and normotensive rats. Measurement of the phosphoinositide distribution in both SHR and WKY indicates that the change observed in /sup 32/P-TPI could not be accounted for by a reduced phosphatidylinositol content in SHR membrane. Measurement of the Mg/sup 2 +/-activated TPI-phosphomonoesterase and of the Ca/sup 2 +/-activated polyphosphoinositide phosphodiesterase activities did not show any significant difference between SHR and WKY. It thus appears that the altered phosphoinositide metabolism observed in hypertensive rats was a consequence of some alteration in the activity of kinases which are responsible for the conversion of phosphatidylinositol into DPI and TPI. These results also suggest that the defect in phosphoinositide metabolism observed in genetically hypertensive rats was not a consequence of the blood pressure elevation and could be related to the pathogenesis of hypertension.

  5. An insulin-sensitive cytosolic protein kinase accounts for the regulation of ATP citrate-lyase phosphorylation.

    PubMed Central

    Yu, K T; Benjamin, W B; Ramakrishna, S; Khalaf, N; Czech, M P

    1990-01-01

    Purified rat liver ATP citrate-lyase is phosphorylated on serine residues by an insulin-stimulated cytosolic kinase activity partially purified from rat adipocytes [Yu, Khalaf & Czech (1987) J. Biol. Chem. 262, 16677-16685]. The Km for lyase phosphorylation by this hormone-sensitive kinase activity is approx. 3 microM. Two-dimensional tryptic-peptide mapping of the 32P-labelled lyase reveals that the kinase-catalysed phosphorylation occurs primarily on a specific peptide. In intact 32P-labelled adipocytes, insulin enhances the serine phosphorylation of ATP citrate-lyase by 2-3-fold. Tryptic digestion of the 32P-labelled lyase immunopurified from insulin-treated adipocytes also yields one major phosphopeptide. 32P-labelled lyase tryptic peptides derived from labelling experiments in vitro and in vivo exhibit identical electrophoretic and chromatographic migration profiles. Furthermore, radio-sequencing of the phosphopeptide from lyase 32P-labelled in vitro indicates that serine-3 from the N-terminus is phosphorylated by the insulin-stimulated cytosolic kinase, in agreement with previous studies on the position of the phosphoserine residue in ATP citrate-lyase isolated from insulin-treated cells. Taken together, the similarity in site-specific phosphorylation of ATP citrate-lyase from insulin-treated adipocytes to that catalysed by the hormone-activated cytosolic kinase in vitro strongly suggests that this kinase mediates insulin action on lyase phosphorylation in intact cells. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:2114095

  6. Pertussis toxin-sensitive G-protein mediates the alpha 2-adrenergic receptor inhibition of melatonin release in photoreceptive chick pineal cell cultures

    SciTech Connect

    Pratt, B.L.; Takahashi, J.S.

    1988-07-01

    The avian pineal gland is a photoreceptive organ that has been shown to contain postjunctional alpha 2-adrenoceptors that inhibit melatonin synthesis and/or release upon receptor activation. Physiological response and (32P)ADP ribosylation experiments were performed to investigate whether pertussis toxin-sensitive guanine nucleotide-binding proteins (G-proteins) were involved in the transduction of the alpha 2-adrenergic signal. For physiological response studies, the effects of pertussis toxin on melatonin release in dissociated cell cultures exposed to norepinephrine were assessed. Pertussis toxin blocked alpha 2-adrenergic receptor-mediated inhibition in a dose-dependent manner. Pertussis toxin-induced blockade appeared to be noncompetitive. One and 10 ng/ml doses of pertussis toxin partially blocked and a 100 ng/ml dose completely blocked norepinephrine-induced inhibition. Pertussis toxin-catalyzed (32P)ADP ribosylation of G-proteins in chick pineal cell membranes was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Membranes were prepared from cells that had been pretreated with 0, 1, 10, or 100 ng/ml pertussis toxin. In the absence of pertussis toxin pretreatment, two major proteins of 40K and 41K mol wt (Mr) were labeled by (32P)NAD. Pertussis toxin pretreatment of pineal cells abolished (32P) radiolabeling of the 40K Mr G-protein in a dose-dependent manner. The norepinephrine-induced inhibition of both cAMP efflux and melatonin release, as assessed by RIA of medium samples collected before membrane preparation, was also blocked in a dose-dependent manner by pertussis toxin. Collectively, these results suggest that a pertussis toxin-sensitive 40K Mr G-protein labeled by (32P)NAD may be functionally associated with alpha 2-adrenergic signal transduction in chick pineal cells.

  7. Water channel proteins: from their discovery in 1985 in Cluj-Napoca, Romania, to the 2003 Nobel Prize in Chemistry.

    PubMed

    Benga, Gh

    2006-10-30

    Water channel proteins, later called aquaporins, are transmembrane proteins that have as their main(specific) function the water transport across biological membranes. The first water channel protein (WCP), now called aquaporin 1, was identified or "seen" in situ (hence discovered) in the human red blood cell (RBC) membrane in 1985 by Benga's group (Cluj-Napoca, Romania). This was achieved by a very selective radiolabeling of RBC membrane proteins with the water transport inhibitor [203Hg]-p-chloromercuribenzene sulfonate (PCMBS), under conditions of specific inhibition. The presence and location of the WCP was discovered among the polypeptides migrating in the region of 35-60 kDa on the electrophoretogram of RBC membrane proteins. The work was first published in 1986 in Biochemistry and Eur. J. Cell Biol. and reviewed by Benga in several articles in 1988-2004. We have thus a world priority in the discovery of the first water channel in the RBC membrane, that was re-discovered by chance by the group of Agre (Baltimore, USA) in 1988, when they isolated a new protein from the RBC membrane, nick-named CHIP28 (channel-forming integral membrane protein of 28 kDa). However, in addition to the 28 kDa component, this protein had a 35-60 kDa glycosylated component, the one detected by Benga's group. Only in 1992 the Agre's group suggested that "it is likely that CHIP28 is a functional unit of membrane water channels". In 1993 CHIP28 was renamed aquaporin 1. Looking in retrospect, asking the crucial question, when was the first WCP, discovered, a fair and clear cut answer would be: the first WCP, now called aquaporin 1, was identified or "seen" (hence discovered) in situ in the human RBC membrane by Benga and coworkers in 1985. It was again "seen" when it was purified in 1988 and again identified when its water transport property was found byAgre's group in 1992. If we make a comparison with the discovery of New World of America, the first man who has "seen" a part, very

  8. Geochemical data for mercury, methylmercury, and other constituents in sediments from Englebright Lake, California, 2002

    USGS Publications Warehouse

    Alpers, Charles N.; Hunerlach, Michael P.; Marvin-DePasquale, Mark C.; Antweiler, Ronald C.; Lasorsa, Brenda K.; De Wild, John F.; Snyder, Noah P.

    2006-01-01

    Deep coring penetrated the full thickness of material deposited after 1940 at six locations in the reservoir; the cores reached a maximum depth of 32.8 meters below the reservoir floor. At the three deep coring sites closest to Englebright Dam, concentrations of HgT (dry basis) were consistently in the range of 100 to 500 ng/g (nanogram per gram), in sediment dominantly of silt size (median grain size of 0.004 to 0.063 mm [millimeter]). At the deep coring sites located farther upstream, the upper parts of the profile had lower concentrations of HgT, generally ranging from 2 to 100 ng/g, in sediment dominantly of sand size (median grain size from 0.063 to 2 mm). The lower part of the vertical profiles at three upstream coring sites had higher concentrations of HgT than the upper and middle parts of these profiles, and had finer median grain size. The highest median concentration of MeHg (1.1 ng/g) was in the top 2 cm (centimeter) of the shallow box cores. This vertical interval also had the highest value of the ratio of MeHg to HgT, 0.41 percent. Median concentrations of MeHg and median values of MeHg/HgT decreased systematically with depth from 0-4 to 4-8 to 8-12 cm in the shallow cores. However, similar systematic decreases were not observed at the meter scale in the deep cores of the MEM (MEthylMercury) series. The overall median of the ratio MeHg/HgT in the deep cores was 0.25 percent, not much less than the overall median value for the shallow cores (0.33 percent). Mercury-203 radiotracer divalent inorganic mercury (203Hg(II)) was used to determine microbial mercury-methylation potential rates for 11 samples collected from three reservoir locations and various depths in the sediment profile. For the five shallow mercury-methylation subsamples, ancillary geochemical parameters were assayed, including microbial sulfate reduction rates, sulfur speciation (sediment acid volatile sulfide, total reduced sulfur, and pore-water sulfate), iron speciation (sediment acid

  9. Two and 8-azido photoaffinity probes. 1. Enzymatic synthesis, characterization, and biological properties of 2- and 8-azido photoprobes of 2-5A and photolabeling of 2-5A binding proteins

    SciTech Connect

    Suhadolnik, R.J.; Kariko, K.; Sobol, R.W. Jr.; Li, S.W.; Reichenbach, N.L.; Haley, B.E.

    1988-11-29

    The 2- and 8-azido trimer 5'-triphosphate photoprobes of 2-5A have been enzymatically synthesized from (..gamma..-/sup 32/P)2-azidoATP and (..cap alpha..-/sup 32/P)8-azidoAPT by 2-5A synthetase from rabbit reticulocyte lysates. Identification and structural determination of the 2- and 8-azido adenylate trimer 5'-triphosphates were accomplished by enzymatic hydrolyses with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase. Hydrolysis products were identified by HPLC and PEI-cellulose TLC analyses. The 8-azido photoprobe of 2-5A displaces p/sub 3/A/sub 4/(/sup 32/P)pCp from RNase L with affinity equivalent to p/sub 3/A/sub 3/. The 8-azido photoprobe also activates RNase L to hydrolyze poly(U)(/sup 32/P)pCp 50% at 7 /times/ 10/sup /minus/9/ M in core-cellulose assays. The 2- and 8-azido photoprobes and authentic p/sub 3/A/sub 3/ activate RNase L to cleave 28S and 18S rRNA to specific cleavage products at 10/sup /minus/9/ M in rRNA cleavage assays. The nucleotide binding site(s) of RNase L and/or other 2-5A binding proteins in extracts of interferon-treated L929 cells were investigated by photoaffinity labeling. Dramatically different photolabeling patterns were observed with the 2- and 8-azido photoprobes. The (..gamma..-/sup 32/P)2-azido adenylate trimer 5'-triphosphate photolabels only one polypeptide with a molecular weight of 185,000 as determined by SDS gel electrophoresis, whereas the (..cap alpha..-/sup 32/P)8-azido adenylate trimer 5'-triphosphate covalently photolabels six polypeptides with molecular weights of 46,000, 63,000, 80,000, 89,000, 109,000, and 158,000. Evidence that the photolabeling by 2- and 8-azido 2-5A photoprobes was highly specific for the p/sub 3/A/sub 3/ allosteric binding site was obtained.

  10. Purification and characterization of two potent heat-stable protein inhibitors of protein phosphatase 2A from bovine kidney.

    PubMed

    Li, M; Guo, H; Damuni, Z

    1995-02-14

    Two heat-stable protein inhibitors of protein phosphatase 2A (PP2A), tentatively designated I1PP2A and I2PP2A, have been purified to apparent homogeneity from extracts of bovine kidney. The purified preparations of I1PP2A exhibited an apparent M(r) approximately 30,000 and 250,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography on Sephacryl S-300, respectively. In contrast, the purified preparations of I2PP2A exhibited an apparent M(r) approximately 20,000 and 80,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel permeation chromatography on Sephacryl S-200, respectively. The purified preparations of I1PP2A and I2PP2A inhibited PP2A with 32P-labeled myelin basic protein, 32P-labeled histone H1, 32P-labeled pyruvate dehydrogenase complex, 32P-labeled phosphorylase, and protamine kinase as substrates. By contrast, I1PP2A and I2PP2A exhibited little effect, if any, on the activity of PP2A with 32P-labeled casein, and did not prevent the autodephosphorylation of PP2A in incubations with the autophosphorylation-activated protein kinase [Guo, H., & Damuni, Z. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 2500-2504]. The purified preparations of I1PP2A and I2PP2A had little effect, if any, on the activities of protein phosphatase 1, protein phosphatase 2B, protein phosphatase 2C, and pyruvate dehydrogenase phosphatase. With 32P-labeled MBP as a substrate, kinetic analysis according to Henderson showed that I1PP2A and I2PP2A were noncompetitive and displayed a Ki of about 30 and 25 nM, respectively. Following cleavage with Staphylococcus aureus V8 protease, I1PP2A and I2PP2A displayed distinct peptide patterns, indicating that these inhibitor proteins are the products of distinct genes. The N-terminal amino acid sequences of the purified preparations indicate that I1PP2A and I2PP2A are novel proteins.

  11. Stirring system for radioactive waste water storage tank

    SciTech Connect

    Ogata, Yoshimune; Nishizawa, Kunihide . Radioisotope Research Center)

    1999-07-01

    A stirring system for 100-m[sup 3] radioactive liquid waste tanks was constructed to unify radioactive concentrations in the tank. The stirring system is effective in certifying that the radioactive concentrations in the tanks are less than the legal limits before they are drained away as waste liquid. This system is composed of discharge units, pipe lines, and a controller. The performance of the system was assessed by comparing the calculated red ink and [sup 32]P concentrations with those monitored at six locations in the tanks. The concentration reached equilibrium after stirring 60 o 120 min with discharge units equipped with six fixed openings configured in differing directions. Residual chlorine in city water used for dilution occasionally bleached the red ink and reduced its concentration. The adsorption of [sup 32]P by slime on the walls of the tanks storing actual waste water lowered the equilibrium concentration.

  12. Lipopolysaccharide (LPS) alters phosphatidylcholine metabolism in elicited peritoneal macrophages

    SciTech Connect

    Grove, R.I.; Allegretto, N.J.; Kiener, P.A.; Warr, G.A. )

    1990-07-01

    We investigated the effects of LPS on mouse peritoneal macrophage phospholipids using radiolabeled precursors. LPS (200 ng/ml) stimulated incorporation of ({sup 32}P) into all classes of phospholipids within 0.5 hr, and after 2 hr the increase was 60% greater than controls. Separation of the phospholipid classes by thin-layer chromatography revealed a selective increase in incorporation of label into phosphatidylcholine (PC) (90% increase compared to approximately 50% in the other phospholipids). In macrophages labeled with ({sup 3}H)-choline, LPS stimulated both the incorporation of label into PC and the release of incorporated label into the medium. The time dependencies of stimulated ({sup 3}H) release and ({sup 32}P) incorporation were similar. These data are consistent with the hypothesis that LPS activates macrophages via a PC-specific phospholipase-dependent mechanism.

  13. Phosphate from the phosphointermediate (EP) of the human red blood cell Na/K pump is coeffluxed with Na, in the absence of external K

    PubMed Central

    1994-01-01

    This study is concerned with Na/K pump-mediated phosphate efflux that occurs during uncoupled Na efflux in human red blood cells. Uncoupled Na efflux is known to be a ouabain-sensitive mode of the Na/K pump that occurs in the absence of external Nao and Ko. Because this efflux (measured with 22Na) is also inhibited by 5 mM Nao, the efflux can be separated into a Nao-sensitive and a Nao-insensitive component. Previous work established that the Nao-sensitive efflux is actually comprised of an electroneutral coefflux of Na with cellular anions, such as SO4 (as 35SO4). The present work focuses on the Nao-insensitive component in which the principal finding is that orthophosphate (P(i)) is coeffluxed with Na in a ouabain-sensitive manner. This P(i) efflux can be seen to occur, in the absence of Ko, in both DIDS-treated intact cells and resealed red cell ghosts. This efflux of P(i) was shown to be derived directly from the pump's substrate, ATP, by the use of resealed ghosts made to contain both ATP and P(i) in which either the ATP or the P(i) were labeled with, respectively, [gamma-32P]ATP or [32P]H3PO4. (These resealed ghosts also contained Na, Mg, P(i), SO4, Ap5A, as well as an arginine kinase/creatine kinase nucleotide regenerating system for the control of ATP and ADP concentrations, and were suspended usually in (NMG)2SO4 at pH 7.4.) It was found that 32P was only coeffluxed with Na when the 32P was contained in [gamma-32P]ATP and not in [32P]H3PO4. This result implies that the 32P that is released comes from ATP via the pump's phosphointermediate (EP) without commingling with the cellular pool of P(i). Ko (as K2SO4) inhibits this 32P efflux as well as the Nao-sensitive 35SO4 efflux, with a K0.5 of 0.3-0.4 mM. The K0.5 for inhibition of P(i) efflux by Ko is not influenced by Nao, nor can Nao act as a congenor for Ko in any of the flux reactions involving Ko. The stoichiometry of Na to SO4 and Na to P(i) efflux is approximately 2:1 under circumstances where the

  14. Oligonucleotides with rapid turnover of the phosphate groups occur endogenously in eukaryotic cells

    SciTech Connect

    Plesner, P.; Goodchild, J.; Kalckar, H.M.; Zamecnik, P.C.

    1987-04-01

    Endogenous oligonucleotides were found in trichloroacetic acid extracts of hamster lung fibroblasts and Tetrahymena cells. Peaks of radioactivity that eluted with retention times similar to oligonucleotide markers (5- to 50-mer) were found by HPLC in cells labeled briefly with /sup 32/Pi. Only minute amounts of UV-absorbing material were detected, consistent with a rapid turnover of phosphate groups. The /sup 32/P-labeled material also migrated as oligonucleotides on 20% polyacrylamide gels; it was not hydrolyzed by alkaline phosphatase but was digested by snake venom phosphodiesterase, S1 nuclease, and pancreatic RNase and was phosphorylated by T4 polynucleotide kinase. The /sup 32/P-labeled material isolated by HPLC was alkali labile and the hydrolyzate ran as nucleotides on paper chromatography. It is concluded that the oligonucleotides are mainly oligoribonucleotides, but it is possible that oligodeoxynucleotides are also present.

  15. New method for determining lecithin and sphingomyelin in amniotic fluid.

    PubMed

    McDonald, L J; Robin, N I; Siegel, L

    1981-03-01

    We describe a new method of complete analysis for lecithin, lysolecithin, and sphingomyelin in amniotic fluid. The analysis is based on alkaline hydrolysis of the lecithins and the enzymic hydrolysis of sphingomyelin. The choline formed in each instance is enzymically phosphorylated with [gamma-32P]ATP to yield [gamma-32P]phosphorylcholine, which is isolated by anion-exchange chromatography and shown to be stoichiometrically related to lecithin, lysolecithin, and sphingomyelin. Other phospholipids do not interfere. Using this methodology, we developed three potential approaches to the assessment of fetal lung maturity: the ratio of lecithin plus lysolecithin (total lecithin) to sphingomyelin, the total lecithin concentration, and total lecithin as a percentage of total amniotic fluid phospholipids. All three indices were compared with the lecithin/sphingomyelin ratio obtained by a chromatographic procedure. Our data suggest that measuring the percentage of total lecithin may provide the best means of evaluating fetal lung maturity, but a final judgment must await clinical investigation.

  16. ADP-ribosylation of membrane components by pertussis and cholera toxin

    SciTech Connect

    Ribeiro-Neto, F.A.P.; Mattera, F.; Hildebrandt, J.D.; Codina, J.; Field, J.B.; Birnbaumer, L.; Sekura, R.D.

    1985-01-01

    Pertussis and cholera toxins are important tools to investigate functional and structural aspects of the stimulatory (N/sub s/) and inhibitory (N/sub i/) regulatory components of adenylyl cyclase. Cholera toxin acts on N/sub s/ by ADP-ribosylating its ..cap alpha../sub s/ subunit; pertussis toxin acts on N/sub i/ by ADP-ribosylating its ..cap alpha..; subunit. By using (/sup 32/P)NAD/sup +/ and determining the transfer of its (/sup 32/P)ADP-ribose moiety to membrane components, it is possible to obtain information on N/sub s/ and N/sub i/. A set of protocols is presented that can be used to study simultaneously and comparatively the susceptibility of N/sub s/ and N/sub i/ to be ADP-ribosylated by cholera and pertussis toxin.

  17. Synthesis and secretion of phosphorylated growth hormone by rat pituitary glands in vitro

    SciTech Connect

    Liberti, J.P.; Joshi, G.S.

    1986-06-13

    Rat anterior pituitary glands were incubated in buffered medium, pH 7.4, containing /sup 32/Pi. After incubation the tissue and medium were separated and a post-mitochondrial supernate (PMS) of the tissue homogenate was prepared. Gel filtration of the PMS and medium resulted in a radioactive peak which coincided with the elution volume of authentic rat growth hormone (rGH). Polyacrylamide gel electrophoresis of the radioactive peak under denaturing condition resulted in a protein-staining band having the same mobility as authentic rGH. Autoradiography of the gels revealed radioactivity precisely at the position of growth hormone as well as elsewhere. The specific radioactivity of the PMS (/sup 32/P)GH was estimated to be 5 to 10 times greater than that of tissue (/sup 32/P)GH. These results indicate that phosphorylated GH is synthesized and secreted by pituitary glands in vitro.

  18. Ethanol stimulates phospholipid turnover and inositol 1,4,5-trisphosphate production in Chlamydomonas eugametos gametes.

    PubMed

    Musgrave, A; Kuin, H; Jongen, M; de Wildt, P; Schuring, F; Klerk, H; van den Ende, H

    1992-02-01

    Alcohols induce mating-structure activation in Chlamydomonas eugametos gametes. From the effect of ethanol on the (32)P-labelling of polyphosphoinositides, we conclude that the synthesis of these lipids is stimulated. Biologically inactive concentrations of ethanol (<6%) had no effect on synthesis, but 6-8% ethanol stimulated synthesis for upto 60 min. The (32)P incorporated into polyphosphoinositides and phosphatidic acid during ethanol treatment was readily chased out when 1 mM unlabelled Na3PO4 was added. Using a binding assay for inositol 1,4,5-trisphosphate, we show that the production of this phospholipid constituent is dramatically increased after ethanol treatment. This effect, coupled to a rise in intracellular calcium concentration, could explain gamete activation. The significance of these results in explaining other ethanol-induced phenomena in algae is discussed.

  19. Orthophosphate turnover in East African lakes.

    PubMed

    Peters, Robert Henry; MacIntyre, Sally

    1976-12-01

    Turnover rates of (32)P-PO4 and concentrations of orthophosphate as soluble reactive phosphorus (SRP) were measured in five East African waters. Rapid incorporation of (32)P-PO4 by the seston and orthophosphate concentrations below the limit of detectibility were found in Lakes Elmenteita, Naivasha, and Naivasha Crater Lake. Turnover was slow and orthophosphate concentration high in both Lake Nakuru and the Crescent Island Crater basin of Lake Naivasha. Further experiments in Lake Nakuru indicated that colloidal binding of orthophosphate was limited and that particles retained by an 8.0 μ filter incorporated 66% as much tracer as particles retained by a 0.1 μ filter. These experiments strengthen our conclusion that a large quantity of orthophosphate is available for algal use in Lake Nakuru.

  20. Chromosomal DNA probes for the identification of Bacteroides tectum and Bacteroides fragilis from the oral cavity of cats.

    PubMed

    Love, D N; Bailey, G D

    1993-01-01

    A dot-blot hybridisation assay using high molecular weight DNA as whole chromosomal probes was used to differentiate Bacteroides tectum from Bacteroides fragilis. 32P-labelled probes were compared with digoxigenin (DIG)-labelled probes. The whole chromosomal probes were specific--differentiating B. tectum from B. fragilis and both from a variety of other species (including other members of the genera Bacteroides, Fusobacterium, Eubacterium, and Prevotella) found in normal and abnormal mouths of cats and horses. However, even at very high stringencies, B. tectum homology groups I, II and III were not distinguishable from one another using either 32P-labelled or DIG-labelled probes. Thus, DIG-labelled whole chromosome probes directed against cellular DNA released directly onto nitrocellulose membranes is considered a useful method for diagnostic veterinary laboratories wishing to identify B. tectum and distinguish it from B. fragilis and other oral anaerobic flora of cats.

  1. Cholera toxin can catalyze ADP-ribosylation of cytoskeletal proteins

    SciTech Connect

    Kaslow, H.R.; Groppi, V.E.; Abood, M.E.; Bourne, H.R.

    1981-11-01

    Cholera toxin catalyzes transfer of radiolabel from (/sup 32/P)NAD/sup +/ to several peptides in particulate preparations of human foreskin fibroblasts. Resolution of these peptides by two-dimensional gel electrophoresis allowed identification of two peptides of M/sub r/ = 42,000 and 52,000 as peptide subunits of a regulatory component of adenylate cyclase. The radiolabeling of another group of peptides (M/sub r/ = 50,000 to 65,000) suggested that cholera toxin could catalyze ADP-ribosylation of cytoskeletal proteins. This suggestion was confirmed by showing that incubation with cholera toxin and (/sup 32/P)NAD/sup +/ caused radiolabeling of purified microtubule and intermediate filament proteins.

  2. Mapping of radiolabeled peptides derived from proteolysis of polypeptides bound to nitrocellulose after Western blotting

    SciTech Connect

    Carrey, E.A.; Hardie, D.G.

    1986-11-01

    Sections of nitrocellulose containing bound /sup 32/P-labeled polypeptides were excised from Western blots and exhaustively digested by trypsin in order to analyze the distribution of phosphorylation sites between the products of limited proteolysis of the multifunctional protein CAD. Using the criterion of analytical isoelectric focusing, the /sup 32/P-peptides obtained by this method were found to be similar, although not identical, to peptides obtained by a more conventional digestion of trichloroacetic acid precipitates. Digestion on Western blots is more straightforward than electrophoretic elution of individual gel slices, gives better recoveries than direct digestion of gel slices, and is particularly suitable for peptide mapping of small peptides which bind to nitrocellulose but would diffuse out of polyacrylamide gels during the commonly used fixing and staining procedures.

  3. Kinetin and Carbohydrate Metabolism in Chinese Cabbage

    PubMed Central

    Berridge, M. V.; Ralph, R. K.

    1971-01-01

    The effects of kinetin on starch and sugar levels and on 14CO2 and 32P-orthophosphate labeling patterns of floated Chinese cabbage (Brassica pekinensis) leaf discs were investigated. Kinetin caused gross starch degradation. Neutral sugars were depressed by 30 to 40% in leaf tissue treated with kinetin for 24 hours. 14CO2 labeling of leaf discs pretreated with kinetin for 24 hours showed increased radioactivity in chloroform-soluble material and most sugar phosphates, and a 35 to 40% decrease in radioactivity in the neutral sugars, glucose, sucrose, and fructose. Incorporation into ATP was increased by 40% by kinetin. 32P-Orthophosphate uptake was inhibited 30% by kinetin. When corrected for uptake, kinetin stimulated incorporation into chloroform-soluble material but had little effect on other cell fractions. These results indicate that kinetin mobilizes starch reserves and increases the flow of sugars required for the synthesis of lipids and structural materials in floated discs. Images PMID:16657661

  4. Synthesis of hyaluronate in differentiated teratocarcinoma cells. Mechanism of chain growth.

    PubMed Central

    Prehm, P

    1983-01-01

    Hyaluronate could be labelled in vivo with [32P]phosphate. [32P]UDP in an alpha-glycosidic linkage constituted the reducing end of membrane-bound hyaluronate. The UDP is liberated during further chain elongation, indicating that chain growth occurs at the reducing end. [3H]Uridine could be incorporated into hyaluronate during synthesis on the isolated membraneous fraction from [3H]UDP-GlcNAc and [3H]UDP-GlcA, confirming the identification of UDP as a constituent of membrane-bound hyaluronate. These results led to a model of hyaluronate chain elongation at the reducing end by alternate addition of the chains to the substrates. Membrane-bound pyrophosphatases or 5'-nucleotidase are suggested as modulators of hyaluronate synthesis. Images Fig. 3. Fig. 4. PMID:6870820

  5. GTP and cytosol stimulate phosphoinositide hydrolysis in isolated platelet membranes

    SciTech Connect

    Baldassare, J.J.; Fisher, G.J.

    1986-06-13

    Hydrolysis of polyphosphoinositides by phospholipase C was examined in isolated membranes prepared from (/sup 32/P)labelled platelets. In the presence of guanosine 5'-(3-O)-thiotriphosphate (GTP..gamma..S), thrombin increased the release of inositol triphosphate and inositol biphosphate approximately 500%. GTP..gamma..S alone stimulated release 2 fold. Maximal activation of thrombin-induced phosphoinositide hydrolysis was observed at 10 ..mu..M GTP. Although addition of calcium had no effect, 2 mM EGTA completely inhibited inositolphosphate release. Addition of high speed supernatant to (/sup 32/P)labelled membranes stimulated the release of inositolphosphates. This hydrolysis was further enhanced by the addition of GTP. These data demonstrate that the breakdown of polyphosphoinositides in isolated platelet membranes is dependent on GTP and stimulated by platelet cytosol.

  6. Onset of cell division in maize germination: action of auxins

    SciTech Connect

    de Jimenez, E.S.; Baiza, A.; Aguilar, R.

    1987-04-01

    Seed germination implies metabolic reactivation, synthesis of macromolecules and onset of cell division. During maize germination, meristematic tissues of embryos re-initiate cell division asynchronically. Since auxins are known to stimulate cell division, they asked how auxins might regulate cell cycle re-initiation. Embryonic tissues were incubated with and without auxins. A pulse of either /sup 3/H-thymidine or /sup 32/P-ortophosphate was given to the tissues. Mitotic indexes were determined and % of labeled mitotic cells recorded. Results indicated that meristematic cells re-initiate cell division either from G/sub 1/ or G/sub 2/ phases. Auxin stimulated differentially the cell division process of these cells. /sup 32/P incorporation into cytoplasmic or nucleic histones was measured. Auxins stimulated this incorporation. Active turnover of histone phosphorylation occurred simultaneously to the cell division process. It is suggested that auxins might regulate the cell cycle by phosphorylation-dephosphorylation of histones.

  7. Phytochrome regulates GTP-binding protein activity in the envelope of pea nuclei

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Memon, A. R.; Thompson, G. A. Jr; Roux, S. J.

    1993-01-01

    Three GTP-binding proteins with apparent molecular masses of 27, 28 and 30 kDa have been detected in isolated nuclei of etiolated pea plumules. After LDS-PAGE and transfer to nitrocellulose these proteins bind [32P]GTP in the presence of excess ATP, suggesting that they are monomeric G proteins. When nuclei are disrupted, three proteins co-purify with the nuclear envelope fraction and are highly enriched in this fraction. The level of [32P]GTP-binding for all three protein bands is significantly increased when harvested pea plumules are irradiated by red light, and this effect is reversed by far-red light. The results indicate that GTP-binding activity associated with the nuclear envelope of plant cells is photoreversibly regulated by the pigment phytochrome.

  8. Accidental neutron dosimetry with human hair

    NASA Astrophysics Data System (ADS)

    Ekendahl, Daniela; Bečková, Věra; Zdychová, Vlasta; Bulánek, Boris; Prouza, Zdeněk; Štefánik, Milan

    2014-11-01

    Human hair contains sulfur, which can be activated by fast neutrons. The 32S(n,p)32P reaction with a threshold of 2.5 MeV was used for fast neutron dose estimation. It is a very important parameter for individual dose reconstruction with regards to the heterogeneity of the neutron transfer to the human body. Samples of human hair were irradiated in a radial channel of a training reactor VR-1. 32P activity in hair was measured both, directly by means of a proportional counter, and as ash dispersed in a liquid scintillator. Based on neutron spectrum estimation, a relationship between the neutron dose and induced activity was derived. The experiment verified the practical feasibility of this dosimetry method in cases of criticality accidents or malevolent acts with nuclear materials.

  9. Phytochrome regulates GTP-binding protein activity in the envelope of pea nuclei

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Memon, A. R.; Thompson, G. A. Jr; Roux, S. J.

    1993-01-01

    Three GTP-binding proteins with apparent molecular masses of 27, 28 and 30 kDa have been detected in isolated nuclei of etiolated pea plumules. After LDS-PAGE and transfer to nitrocellulose these proteins bind [32P]GTP in the presence of excess ATP, suggesting that they are monomeric G proteins. When nuclei are disrupted, three proteins co-purify with the nuclear envelope fraction and are highly enriched in this fraction. The level of [32P]GTP-binding for all three protein bands is significantly increased when harvested pea plumules are irradiated by red light, and this effect is reversed by far-red light. The results indicate that GTP-binding activity associated with the nuclear envelope of plant cells is photoreversibly regulated by the pigment phytochrome.

  10. Dinucleotide repeat polymorphisms in waterfowl (family Anatidae): Characterization of a sex-linked (Z-specific) and 14 autosomal loci

    USGS Publications Warehouse

    Buchholz, W.G.; Pearce, J.M.; Pierson, B.J.; Scribner, K.T.

    1998-01-01

    Canada goose (Branta Canadensis) and harlequin duck (Histrionicus histrionicus) DNAs were digested with Sau3AI, and size selected (300-700 bp) fragments were ligated into BamHI-digested pBluscriptII KS+. The enrichment protocol of Ostrander et al.1 was followed. The resulting libraries were screened using a [ƴ-32P]ATP end-labelled (CA)20 oligonucleotides as a hybridization probe. Positive clones were sequenced using cycle-sequencing protocols (Epicentre Technologies, Madison, WI) and primers flanking the inserts. PCR primers were designed to amplify the repeat and yield amplification products of ≈100-200 bp. DNA  samples were screened for variation at these loci using [ƴ-32P]ATP end-labelled primers. The products were resolved using 6% denaturing polyacrylamide gels and autoradiography.

  11. Phospholipid turnover in Torpedo marmorata electric organ during discharge in vivo.

    PubMed Central

    Bleasdale, J E; Hawthorne, J N; Widlund, L; Heilbronn, E

    1976-01-01

    One electric organ of anaesthetized Torpedo marmorata was stimulated through electrodes placed on the electric lobe of the brain. Nerves to the other electric organ were cut to provide an unstimulated control. Glucose 6-[32P]phosphate was injected into each organ 16h before electrical stimulation. After stimulation for 10 min at 5 Hz, the organs were removed homogenized and centrifuged on a density gradient for the preparation of subcellular fractions. Stimulation increased the incorporation of 32P into phosphatidate, phosphatidylinositol and phosphatidylcholine. The increased phosphatidate labelling, but not that of the other two lipids, was seen in fractions rich in synaptic vesicles. Stimulation had no effect on ATP labelling. The phosphatidate content of most fractions fell slightly after stimulation, but amounts of other phospholipids were not affected. Images PLATE 1 PLATE 2 PMID:825114

  12. Studies of endogenous UDP-glucose: glycoprotein glucose-1-phosphotransferase acceptors in embryonic chick neural tissues

    SciTech Connect

    Hiller, A.M.S.

    1986-01-01

    UPD-glucose: glycoprotein glucose-1-phosphotransferase (GlcPTase) catalyzes the transfer of a Glc-1-P from UDP-Glc to high mannose-type oligosaccharides on a class of embryonic chick neutral retinal glycoproteins. The primary acceptors for this glycosylphosphotransferase were determined by incubating embryonic chick tissue sonicates with (B-/sup 32/P)UDP-Glc followed by SDS-polyacrylamine gel electrophoresis and autoradiography. The primary GlcPT ase acceptors in brain tissue have apparent subunit molecular weights of 66 and 62 kilodaltons (kd). /sup 32/P incorporation into these peptides was eliminated by excess levels of unlabeled UDP-Glc and unaffected by the inclusion of other phosphate-containing compounds, with the exception of UDP-galactose which can be converted to UDP-Glc by an endogenous epimerase.

  13. Atmospheric Transmission Measurements at White Sands Missile Range, August 1978.

    DTIC Science & Technology

    1980-10-29

    oeay md Identify by black -. b-,) Atin usp Ii ric iran sm 155111 Laser proIpagat ion Iligh-resolut ion Fourier-tranlIrl Spiect roscopy 20 ABSTRACT... NRI . RILPfRt 8422 1.0 1.0 21IP7 3-2P3 1-OPIO 21lP6 3-2P2 14org 0 2 20.. 0.1 j I0.1 2M6.0 t ~ 2662. 2675.0 2675.0 Icm’ 1 2687. 2700.0 1.0 1.0 -- - 2-1...air Ie)mpe.r.Iiure % h 1 1 4 cm 1 yii % Ilstiqflr)ClI IWJIlItln 0 .11t,11i(1101 26 NRI R1 P()RI 1 422 PATH SELECTION Selection of’ a suitable path for

  14. Restoration by T4 ligase of DNA sequences sensitive to "flush" cleaving restriction enzyme.

    PubMed

    Mottes, M; Morandi, C; Cremaschi, S; Sgaramella, V

    1977-07-01

    Fouteen "flush"-ended segments originate from the action of the restriction endonuclease Hae III of Haemophilus aegiptius on the DNA of the colicinogenic factor ColE 1 (A. Oka and M. Takanami, Nature, 264, 191, 1976). They are joined by the T4 polynucleotide ligase. The reaction can be monitored by gel electrophoresis, electron microscopy and resistance to phosphatase of the 5'-32P labelled ends. The joined products are a random recombination of the original segments, and can be cleaved by the same Hae III endonuclease to restore the exact electrophoretic pattern of the Hae III-cut ColE 1 DNA. In a properly diluted mixture of 5'-32P segments treated with T4 ligase, the level of phosphatase resistance is very close to the frequency of circle-formation as determined by electron microscopy: thus, the joining of the "flush"-ends involves the formation of circular structures covalently closed in both strands.

  15. Catalytic sites of Escherichia coli F sub 1 -ATPase. Characterization of unisite catalysis at varied pH

    SciTech Connect

    Alshawi, M.K.; Senior, A.E. )

    1992-01-28

    Using manual rapid-mixing procedures in which small, equal volumes of Escherichia coli F{sub 1}-ATPase and ({gamma}-{sup 32}P)ATP were combined at final concentrations of 2 and 0.2 {mu}M, respectively it was shown that {ge}66% of the {sup 32}P became bound to the enzyme, with the ratio of bound ATP/bound P{sub i} equal to 0.4 and the rate of dissociation of bound ({sup 32}P)P{sub i} equal to 3.5 {times} 10{sup {minus}3} s{sup {minus}1}, similar to previously published values. In the presence of 1 mM sodium azide, 99% of the {sup 32}P became bound to the enzyme, with the ratio of bound ATP/bound P{sub i} being 0.57. These experiments demonstrated that when conditions are used which minimize cooperative catalysis, most or all of the F{sub 1} molecules bind substoichiometric ATP tightly, hydrolyze it with retention of bound ATP and P{sub i}, and release the products slowly. The data justify the validity of previously published rate constants for unisite catalysis. Unisite catalysis in E. coli F{sub 1}-ATPase was studied at varied pH from 5.5 to 9.5 using buffers devoid of phosphate. The data confirm the view that, during normal oxidative phosphorylation, energy input from {Delta}{mu}{sub H+} is required for P{sub i} binding to ATP release. The data also support the idea that two major enzyme conformations are involved in unisite catalysis and that an ionic interaction influences binding of P{sub i} and release of ATP in one conformation.

  16. ATP binding/hydrolysis by and phosphorylation of peroxisomal ATP-binding cassette proteins PMP70 (ABCD3) and adrenoleukodystrophy protein (ABCD1).

    PubMed

    Tanaka, Arowu R; Tanabe, Kouichi; Morita, Masashi; Kurisu, Mikinori; Kasiwayama, Yoshinori; Matsuo, Michinori; Kioka, Noriyuki; Amachi, Teruo; Imanaka, Tsuneo; Ueda, Kazumitsu

    2002-10-18

    The 70-kDa peroxisomal membrane protein (PMP70) and adrenoleukodystrophy protein (ALDP), half-size ATP-binding cassette transporters, are involved in metabolic transport of long and very long chain fatty acids into peroxisomes. We examined the interaction of peroxisomal ATP-binding cassette transporters with ATP using rat liver peroxisomes. PMP70 was photoaffinity-labeled at similar efficiencies with 8-azido-[alpha-32P]ATP and 8-azido-[gamma-32P]ATP when peroxisomes were incubated with these nucleotides at 37 degrees C in the absence Mg2+ and exposed to UV light without removing unbound nucleotides. The photoaffinity-labeled PMP70 and ALDP were co-immunoprecipitated together with other peroxisomal proteins, which also showed tight ATP binding properties. Addition of Mg2+ reduced the photoaffinity labeling of PMP70 with 8-azido-[gamma-32P]ATP by 70%, whereas it reduced photoaffinity labeling with 8-azido-[alpha-32P]ATP by only 20%. However, two-thirds of nucleotide (probably ADP) was dissociated during removal of unbound nucleotides. These results suggest that ATP binds to PMP70 tightly in the absence of Mg2+, the bound ATP is hydrolyzed to ADP in the presence of Mg2+, and the produced ADP is dissociated from PMP70, which allows ATP hydrolysis turnover. Properties of photoaffinity labeling of ALDP were essentially similar to those of PMP70. Vanadate-induced nucleotide trapping in PMP70 and ALDP was not observed. PMP70 and ALDP were also phosphorylated at a tyrosine residue(s). ATP binding/hydrolysis by and phosphorylation of PMP70 and ALDP are involved in the regulation of fatty acid transport into peroxisomes.

  17. Genotyping of Burkholderia mallei: Effective Subspecies Discrimination Using Ribotyping and Repeat Sequences

    DTIC Science & Technology

    2005-10-01

    Hamsters or mice are the most common laboratory models ! Mortality is very high ! There is no vaccine ! Antibiotics are typically only partially effective...antibiotic resistance "Historically high mortality, no vaccine and relatively poor treatment options "CDC Category B Bioterrorism agent http...transferred and probed with 32P-labeled 18-mer derived from E. coli rDNA (gct cct agt acg aga gga) • Objective is to identify restriction fragment

  18. Materials Growth for InAs High Electron Mobility Transistors and Circuits

    DTIC Science & Technology

    2004-03-01

    operating at room temperature. In this application, the room- temperature mobility and carrier density need to be high ~i.e., low sheet resistance !. In some...density and mobility at 300 K; sheet resistance calculated from transport measurements; sheet resistance measured by Lehighton; and wafer nonuniformity...A typical sheet - resistance map from the 32P MBE ~50 FIG. 4. STEM image ~350 0003! of sample B after gate metallization. FIG. 5. Sheet - resistance map

  19. Peritoneoscintigraphy in detection of improper placement of peritoneal catheter into bowel lumen prior to chromic phosphate P-32 therapy. A case report

    SciTech Connect

    Neutze, J.; Van Nostrand, D.; Major, W.

    1985-11-01

    Radionuclide peritoneoscintigraphy has been used prior to chromic phosphate P-32 (P-32CP) intraperitoneal therapy to assure proper placement of the catheter in the peritoneal cavity, to exclude loculation, and to predict inadequate distribution of P-32CP. This is a case report of the detection of a peritoneal catheter improperly placed into the bowel lumen by pretherapy radionuclide peritoneoscintigraphy, and this case demonstrates the distinguishing characteristics of the radiocolloid distribution secondary to an intraluminal injection relative to an intraperitoneal injection.

  20. Plant oligoadenylates: enzymatic synthesis, isolation, and biological activities

    SciTech Connect

    Devash, Y.; Reichman, M.; Sela, I.; Reichenbach, N.L.; Suhadolnik, R.J.

    1985-01-29

    An enzyme that converts (/sup 3/H, /sup 32/P)ATP, with a /sup 3/H:/sup 32/P ratio of 1:1, to oligoadenylates with the same /sup 3/H:/sup 32/P ratio was increased in plants following treatment with human leukocyte interferon or plant antiviral factor or inoculation with tobacco mosaic virus. The enzyme was extracted from tobacco leaves, callus tissue cultures, or cell suspension cultures. The enzyme, a putative plant oligoadenylate synthetase, was immobilized on poly(rI) . poly(rC)-agarose columns and converted ATP into plant oligoadenylates. These oligoadenylates were displaced from DEAE-cellulose columns with 350 mM KCl buffer, dialyzed, and further purified by high-performance liquid chromatography (HPLC) and DEAE-cellulose gradient chromatography. In all steps of purification, the ratio of /sup 3/H:/sup 32/P in the oligoadenylates remained 1:1. The plant oligoadenylates isolated by displacement with 350 mM KCl had a molecular weight greater than 1000. The plant oligoadenylates had charges of 5- and 6-. HPLC resolved five peaks, three of which inhibited protein synthesis in reticulocyte and wheat germ systems. Partial structural elucidation of the plant oligoadenylates has been determined by enzymatic and chemical treatments. An adenylate with a 3',5'-phosphodiester and/or a pyrophosphoryl linkage with either 3'- or 5'-terminal phosphates is postulated on the basis of treatment of the oligoadenylates with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase and acid and alkaline hydrolyses. The plant oligoadenylates at 8 X 10(-7) M inhibit protein synthesis by 75% in lysates from rabbit reticulocytes and 45% in wheat germ cell-free systems.

  1. Regulatory Biochemical and Metabolic Responses in Photoreceptors.

    DTIC Science & Technology

    1987-11-01

    34𔃾. P• ° *" 19. Abstract (continued) ingle channel activity . These results suggest that opsin , in addition to performing ~its function as the receptor...clamp recording to demonstrate that opsin exhibits cGMP-dependent single channel activity . Two elementary conecli1tances, of 17 and 32 pS, were...recordings of cGMP- activated single channel currents observed in patches excised from phospholipid vesicles containing purified (>99%) bovine opsin

  2. Ion-exchange chromatography separates activities synthesizing and degrading fructose 2,6-bisphosphate from C3 and C4 leaves but not from rat liver

    NASA Technical Reports Server (NTRS)

    Macdonald, F. D.; Chou, Q.; Buchanan, B. B.

    1987-01-01

    Fructose-6-phosphate,2-kinase and fructose-2,6-bisphosphatase were separated on the basis of charge from leaves of C3 (spinach, lettuce, and pea) and C4 (sorghum and amaranthus) plants but not from rat liver--a tissue known to contain a bifunctional enzyme with both activities. [2-32P]Fructose 2,6-bisphosphate binding experiments also suggest that the major forms of these activities reside on different proteins in leaves.

  3. NADP/sup +/ enhances cholera and pertussis toxin-catalyzed ADP-ribosylation of membrane proteins

    SciTech Connect

    Kawai, Y.; Whitsel, C.; Arinze, I.J.

    1986-05-01

    Cholera or pertussis toxin-catalyzed (/sup 32/P)ADP-ribosylation is frequently used to estimate the concentration of the stimulatory (Ns) or inhibitory (Ni) guanine nucleotide regulatory proteins which modulate the activity of adenylate cyclase. With this assay, however, the degradation of the substrate, NAD/sup +/, by endogenous enzymes such as NAD/sup +/-glycohydrolase (NADase) present in the test membranes can influence the results. In this study the authors show that both cholera and pertussis toxin-catalyzed (/sup 32/P)ADP-ribosylation of liver membrane proteins is markedly enhanced by NADP/sup +/. The effect is concentration dependent; with 20 ..mu..M (/sup 32/P)NAD/sup +/ as substrate maximal enhancement is obtained at 0.5-1.0 mM NADP/sup +/. The enhancement of (/sup 32/P)ADP-ribosylation by NADP/sup +/ was much greater than that by other known effectors such as Mg/sup 2 +/, phosphate or isoniazid. The effect of NADP/sup +/ on ADP-ribosylation may occur by inhibition of the degradation of NAD/sup +/ probably by acting as an alternate substrate for NADase. Among inhibitors tested (NADP/sup +/, isoniazid, imidazole, nicotinamide, L-Arg-methyl-ester and HgCl/sub 2/) to suppress NADase activity, NADP/sup +/ was the most effective and, 10 mM, inhibited activity of the enzyme by about 90%. In membranes which contain substantial activities of NADase the inclusion of NADP/sup +/ in the assay is necessary to obtain maximal ADP-ribosylation.

  4. Simultaneous demonstrations of neuropeptide Y gene expression and peptide storage in single neurons of the human brain

    SciTech Connect

    Chan-Palay, V.; Yasargil, G.; Hamid, Q.; Polak, J.M.; Palay, S.L.

    1988-05-01

    A combination of in situ hybridization for neuropeptide Y mRNA that used a /sup 32/P-labeled complementary RNA probe and immunocytochemistry with polyclonal antibodies against neuropeptide Y were applied to human cortical brain samples to simultaneously localize neuropeptide Y and its mRNA. These two techniques allowed simultaneous identification of neuropeptide Y gene expression and peptide storage in single neutrons of the human brain.

  5. Studies on the Biodisposition of Organophosphates in Mice

    DTIC Science & Technology

    1983-08-01

    Biochem. Pharmacol. 19:865-876, 1970. * 8. Ramachandran , B.V.: Distribution of DF32P in mouse organs- I. The effect of route of administration on...Incubation time course for brain, diaphragm and plasma cholinesterase activity in mice and rats 7 2. Effect of DFP on cholinesterase activity 7 0 3...incubations for diaphragm. The major difference between mouse and rat cholinesterase is the higher rate of hydrolysis in the mouse tissues. 2. Effect of

  6. Contingency Base Camp Solid Waste Generation

    DTIC Science & Technology

    2013-09-01

    4. Table 3-2, p. 3-14: This table presents planning factors for components of solid waste assuming none is being reused or recycled. Table 2 lists...combatants. Table 3 lists the comprehensive base camp waste characterization that was developed based on the evaluations of this study. The Wastewater ...Treat- ment Plant (WWTP) sludge production was based on studies at wastewater treatment plants at three base camps. Note that the most sig- nificant

  7. Plant oligoadenylates: enzymatic synthesis, isolation, and biological activities.

    PubMed

    Devash, Y; Reichman, M; Sela, I; Reichenbach, N L; Suhadolnik, R J

    1985-01-29

    An enzyme that converts [3H, 32P]ATP, with a 3H:32P ratio of 1:1, to oligoadenylates with the same 3H:32P ratio was increased in plants following treatment with human leukocyte interferon or plant antiviral factor or inoculation with tobacco mosaic virus. The enzyme was extracted from tobacco leaves, callus tissue cultures, or cell suspension cultures. The enzyme, a putative plant oligoadenylate synthetase, was immobilized on poly(rI) . poly(rC)-agarose columns and converted ATP into plant oligoadenylates. These oligoadenylates were displaced from DEAE-cellulose columns with 350 mM KCl buffer, dialyzed, and further purified by high-performance liquid chromatography (HPLC) and DEAE-cellulose gradient chromatography. In all steps of purification, the ratio of 3H:32P in the oligoadenylates remained 1:1. The plant oligoadenylates isolated by displacement with 350 mM KCl had a molecular weight greater than 1000. The plant oligoadenylates had charges of 5- and 6-. HPLC resolved five peaks, three of which inhibited protein synthesis in reticulocyte and wheat germ systems. Partial structural elucidation of the plant oligoadenylates has been determined by enzymatic and chemical treatments. An adenylate with a 3',5'-phosphodiester and/or a pyrophosphoryl linkage with either 3'- or 5'-terminal phosphates is postulated on the basis of treatment of the oligoadenylates with T2 RNase, snake venom phosphodiesterase, and bacterial alkaline phosphatase and acid and alkaline hydrolyses. The plant oligoadenylates at 8 X 10(-7) M inhibit protein synthesis by 75% in lysates from rabbit reticulocytes and 45% in wheat germ cell-free systems.(ABSTRACT TRUNCATED AT 250 WORDS)

  8. The GTP-binding protein RhoA localizes to the cortical granules of Strongylocentrotus purpuratas sea urchin egg and is secreted during fertilization.

    PubMed

    Cuéllar-Mata, P; Martínez-Cadena, G; López-Godínez, J; Obregón, A; García-Soto, J

    2000-02-01

    The sea urchin egg has thousands of secretory vesicles known as cortical granules. Upon fertilization, these vesicles undergo a Ca2+-dependent exocytosis. G-protein-linked mechanisms may take place during the egg activation. In somatic cells from mammals, GTP-binding proteins of the Rho family regulate a number of cellular processes, including organization of the actin cytoskeleton. We report here that a crude membrane fraction from homogenates of Strongylocentrotus purpuratus sea urchin eggs, incubated with C3 (which ADP-ribosylates specifically Rho proteins) and [32P]NAD, displayed an [32P]ADP-ribosylated protein of 25 kDa that had the following characteristics: i) identical electrophoretic mobility in SDS-PAGE gels as the [32P]ADP-ribosylated Rho from sea urchin sperm; ii) identical mobility in isoelectro focusing gels as human RhoA; iii) positive cross-reactivity by immunoblotting with an antibody against mammalian RhoA. Thus, unfertilized S. purpuratus eggs contain a mammalian RhoA-like protein. Immunocytochemical analyses indicated that RhoA was localized preferentially to the cortical granules; this was confirmed by experiments of [32P]ADP-ribosylation with C3 in isolated cortical granules. Rho was secreted and retained in the fertilization membrane after insemination or activation with A23187. It was observed that the Rho protein present in the sea urchin sperm acrosome was also secreted during the exocytotic acrosome reaction. Thus, Rho could participate in those processes related to the cortical granules, i.e., in the Ca2+-regulated exocytosis or actin reorganization that accompany the egg activation.

  9. The casein kinases Yck1p and Yck2p act in the secretory pathway, in part, by regulating the Rab exchange factor Sec2p

    PubMed Central

    Stalder, Danièle; Novick, Peter J.

    2016-01-01

    Sec2p is a guanine nucleotide exchange factor that activates Sec4p, the final Rab GTPase of the yeast secretory pathway. Sec2p is recruited to secretory vesicles by the upstream Rab Ypt32p acting in concert with phosphatidylinositol-4-phosphate (PI(4)P). Sec2p also binds to the Sec4p effector Sec15p, yet Ypt32p and Sec15p compete against each other for binding to Sec2p. We report here that the redundant casein kinases Yck1p and Yck2p phosphorylate sites within the Ypt32p/Sec15p binding region and in doing so promote binding to Sec15p and inhibit binding to Ypt32p. We show that Yck2p binds to the autoinhibitory domain of Sec2p, adjacent to the PI(4)P binding site, and that addition of PI(4)P inhibits Sec2p phosphorylation by Yck2p. Loss of Yck1p and Yck2p function leads to accumulation of an intracellular pool of the secreted glucanase Bgl2p, as well as to accumulation of Golgi-related structures in the cytoplasm. We propose that Sec2p is phosphorylated after it has been recruited to secretory vesicles and the level of PI(4)P has been reduced. This promotes Sec2p function by stimulating its interaction with Sec15p. Finally, Sec2p is dephosphorylated very late in the exocytic reaction to facilitate recycling. PMID:26700316

  10. Identification of New Substrates for Breast Tumor-Specific LMW Cyclin E/CDk2 Kinase

    DTIC Science & Technology

    2011-09-01

    induced breast cancer and that CDK2 inhibitor (such as roscovitine ) also delays mammary tumor formation (Akli et al., 2011). The hypothesis is that...in the presence of 32P-γ-ATP and with or without roscovitine . The samples were separated by SDS-PAGE and exposed to x-ray films. (E) Schematic of...phosphorylate Hbo1 at relatively similar levels, and addition of roscovitine efficiently inhibited the Hbo1 phosphorylation signal (Figure 2C). Similar

  11. Angiotensin II induces phosphatidic acid formation in neonatal rat cardiac fibroblasts: evaluation of the roles of phospholipases C and D.

    PubMed

    Booz, G W; Taher, M M; Baker, K M; Singer, H A

    1994-12-21

    Phosphatidic acid has been proposed to contribute to the mitogenic actions of various growth factors. In 32P-labeled neonatal rat cardiac fibroblasts, 100 nM [Sar1]angiotensin II was shown to rapidly induce formation of 32P-phosphatidic acid. Levels peaked at 5 min (1.5-fold above control), but were partially sustained over 2 h. Phospholipase D contributed in part to phosphatidic acid formation, as 32P- or 3H-phosphatidylethanol was produced when cells labeled with [32P]H3PO4 or 1-O-[1,2- 3H]hexadecyl-2-lyso-sn-glycero-3-phosphocholine were stimulated in the presence of 1% ethanol. [Sar1]angiotensin II-induced phospholipase D activity was transient and mainly mediated through protein kinase C (PKC), since PKC downregulation reduced phosphatidylethanol formation by 68%. Residual activity may have been due to increased intracellular Ca2+, as ionomycin also activated phospholipase D in PKC-depleted cells. Phospholipase D did not fully account for [Sar1]angiotensin II-induced phosphatidic acid: 1) compared to PMA, a potent activator of phospholipase D, [Sar1]angiotensin II produced more phosphatidic acid relative to phosphatidylethanol, and 2) PKC downregulation did not affect [Sar1]angiotensin II-induced phosphatidic acid formation. The diacylglycerol kinase inhibitor R59949 depressed [Sar1]angiotensin II-induced phosphatidic acid formation by only 21%, indicating that activation of a phospholipase C and diacylglycerol kinase also can not account for the bulk of phosphatidic acid. Thus, additional pathways not involving phospholipases C and D, such as de novo synthesis, may contribute to [Sar1]angiotensin II-induced phosphatidic acid in these cells. Finally, as previously shown for [Sar1]angiotensin II, phosphatidic acid stimulated mitogen activated protein (MAP) kinase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

  12. Lipid A binding sites in membranes of macrophage tumor cells

    SciTech Connect

    Hampton, R.Y.; Golenbock, D.T.; Raetz, C.R.

    1988-10-15

    Lipopolysaccharide affects a variety of eukaryotic cells and mammalian organisms. These actions are involved in the pathogenesis of Gram-negative septicemia. Many of the actions of lipopolysaccharide are believed to be caused by its active moiety, lipid A. Our laboratory has previously identified a bioactive lipid A precursor, termed lipid IVA, which can be labeled with 32P of high specific activity and purified. In this work we have used the labeled probe, 4'-32P-lipid IVA, to develop a novel assay for the specific binding of lipid IVA to whole cells. We have also demonstrated its use in a ligand blotting assay of immobilized cellular proteins. Using the whole cell assay, we show that 4'-32P-lipid IVA specifically binds to RAW 264.7 macrophage-like cultured cells. The binding is saturable, is inhibited with excess unlabeled lipid IVA, and is proteinase K-sensitive. It displays cellular and pharmacological specificity. Using the ligand blotting assay, we show that several RAW 264.7 cell proteins can bind 4'-32P-lipid IVA. The two principal binding proteins have Mr values of 31 and 95 kDa, as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Fractionation studies indicate that the 31-kDa protein is enriched in the nuclear fraction and may be a histone, whereas the 95-kDa protein is enriched in the membrane fraction. The binding assays that we have developed should lead to a clearer understanding of lipid A/animal cell interactions.

  13. IDENTIFICATION OF STEREOCHEMICAL CONFIGURATIONS OF CYCLOPENTA[CD]PYRENE-DNA ADDUCTS IN STRAIN A/J MOUSE LUNG AND C3H10T1/2CL8 CELLS

    EPA Science Inventory

    Identification of Sterochemical Configurations of Cyclopent A[cd]Pyrene DNA Adducts in Strain A/J Mouse Lung and C3H10T1/2CL8 Cells.

    Four major and several minor DNA adducts were resolved by 32P-postlabeling analysis of DNA from strain A/J mouse lung and C3H10T1/2CL8 (C3H...

  14. Purification and characterization of a phosphotyrosyl-protein phosphatase from wheat seedlings.

    PubMed

    Cheng, H F; Tao, M

    1989-10-19

    A neutral phosphatase which catalyzes the hydrolysis of p-nitrophenylphosphate has been purified to homogeneity from wheat seedlings. The enzyme is a monomeric glycoprotein exhibiting a molecular weight of 35,000, frictional ratio of 1.22, Stokes' radius of 260 nm, and sedimentation coefficient of 3.2 S. That the enzyme is a glycoprotein is surmised from its chromatographic property on Concanavalin A-Sepharose column. An examination of the substrate specificity indicates that the enzyme exhibits a preference for phosphotyrosine over a number of phosphocompounds, including p-nitrophenylphosphate and several glycolytic intermediates. Both phosphoserine and phosphothreonine are not hydrolyzed by the enzyme. The phosphatase activity is not affected by high concentrations of chelating agents and does not require metal ions. Molybdate, orthovanadate, Zn2+, and Hg2+ are all potent inhibitors of the phosphatase activity. The ability of the phosphatase to dephosphorylate protein phosphotyrosine has been investigated. [32P-Tyr]poly(Glu,Tyr)n, [32P-Tyr]alkylated bovine serum albumin, [32P-Tyr]angiotensin-I, and [32P-Tyr]band 3 (from human erythrocyte) are all substrates of the phosphatase. On the other hand, the enzyme has no activity toward protein phosphoserine and phosphothreonine. Our result further indicates that the neutral phosphatase is distinct from the wheat germ acid phosphatase. The latter enzyme is found to dephosphorylate phosphotyrosyl as well as phosphoseryl and phosphothreonyl groups in proteins. In light of the many similarities in properties to phosphotyrosyl protein phosphatases isolated from several sources, it is suggested that the wheat seedling phosphatase may participate in cellular regulation involving protein tyrosine phosphorylation.

  15. IDENTIFICATION OF STEREOCHEMICAL CONFIGURATIONS OF CYCLOPENTA[CD]PYRENE-DNA ADDUCTS IN STRAIN A/J MOUSE LUNG AND C3H10T1/2CL8 CELLS

    EPA Science Inventory

    Identification of Sterochemical Configurations of Cyclopent A[cd]Pyrene DNA Adducts in Strain A/J Mouse Lung and C3H10T1/2CL8 Cells.

    Four major and several minor DNA adducts were resolved by 32P-postlabeling analysis of DNA from strain A/J mouse lung and C3H10T1/2CL8 (C3H...

  16. Intra-articular chromic phosphate (³²P) in the treatment of diffuse pigmented villonodular synovitis.

    PubMed

    Zook, Jennifer E; Wurtz, Daniel L; Cummings, Judd E; Cárdenes, Higinia R

    2011-01-01

    Pigmented villonodular synovitis (PVNS) is an uncommon proliferative lesion of synovial tissue. In diffuse PVNS, recurrence rates are high after resection alone. Adjuvant external beam radiation therapy contributes to improved local control. Limited data exist for intra-articular radioisotope therapy after surgical resection. We report institutional experience with intra-articular chromic phosphate ((32)P). Records were reviewed from the Department of Radiation Oncology at Indiana University. Nine cases of PVNS treated with (32)P were identified (mean age=40). Seven patients were treated at time of recurrence and 2 patients were treated prophylactically. Intra-articular injections were performed by accessing the joint space, aspirating joint fluid, reinjecting 1-2 mCi of (32)P, and barbitaging to ensure good distribution in the joint space. No external beam radiation therapy was delivered. One patient was lost to followup. Mean followup of remaining patients was 20 months (range, 2-48). Eleven injections were performed in 9 patients. Eight had PVNS in the knee(s) and 1 patient had involvement of the hip. Two patients were treated more than once, one for a recurrence in the same joint at 13 months and another for PVNS of a contralateral joint. Three clinical recurrences (2, 13, and 28 months) were noted. Two of three recurrences were in patients who had bulky diffuse PVNS at the time of injection. Overall local control was 70%. In patients without bulky diffuse PVNS at the time of injection, local control was 88%. We report success using intra-articular injections of (32)P after synovectomy in patients with PVNS. Copyright © 2011 American Brachytherapy Society. Published by Elsevier Inc. All rights reserved.

  17. Ion-exchange chromatography separates activities synthesizing and degrading fructose 2,6-bisphosphate from C3 and C4 leaves but not from rat liver

    NASA Technical Reports Server (NTRS)

    Macdonald, F. D.; Chou, Q.; Buchanan, B. B.

    1987-01-01

    Fructose-6-phosphate,2-kinase and fructose-2,6-bisphosphatase were separated on the basis of charge from leaves of C3 (spinach, lettuce, and pea) and C4 (sorghum and amaranthus) plants but not from rat liver--a tissue known to contain a bifunctional enzyme with both activities. [2-32P]Fructose 2,6-bisphosphate binding experiments also suggest that the major forms of these activities reside on different proteins in leaves.

  18. Ca(2+)-calmodulin-dependent phosphorylation of islet secretory granule proteins

    SciTech Connect

    Watkins, D.T. )

    1991-08-01

    The effect of Ca2+ and calmodulin on phosphorylation of islet secretory granule proteins was studied. Secretory granules were incubated in a phosphorylation reaction mixture containing (32P)ATP and test reagents. The 32P-labeled proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the 32P content was visualized by autoradiography, and the relative intensities of specific bands were quantitated. When the reaction mixture contained EGTA and no added Ca2+, 32P was incorporated into two proteins with molecular weights of 45,000 and 13,000. When 10(-4) M Ca2+ was added without EGTA, two additional proteins (58,000 and 48,000 Mr) were phosphorylated, and the 13,000-Mr protein was absent. The addition of 2.4 microM calmodulin markedly enhanced the phosphorylation of the 58,000- and 48,000-Mr proteins and resulted in the phosphorylation of a major protein whose molecular weight (64,000 Mr) is identical to that of one of the calmodulin binding proteins located on the granule surface. Calmodulin had no effect on phosphorylation in the absence of Ca2+ but was effective in the presence of calcium between 10 nM and 50 microM. Trifluoperazine and calmidazolium, calmodulin antagonists, produced a dose-dependent inhibition of the calmodulin effect. 12-O-tetradecanoylphorbol 13-acetate, a phorbol ester that activates protein kinase C, produced no increase in phosphorylation, and 1-(5-isoquinoline sulfonyl)-2-methyl piperazine dihydrochloride, an inhibitor of protein kinase C, had no effect. These results indicate that Ca(2+)-calmodulin-dependent protein kinases and endogenous substrates are present in islet secretory granules.

  19. Platelet-derived growth factor induces phosphorylation of a 64-kDa nuclear protein

    SciTech Connect

    Shawver, L.K.; Pierce, G.F.; Kawahara, R.S.; Deuel, T.F.

    1989-01-15

    The platelet-derived growth factor (PDGF) stimulated the phosphorylation of a nuclear protein of 64 kDa (pp64) in nuclei of nontransformed normal rat kidney (NRK) cells. Low levels of phosphorylation of pp64 were observed in nuclei of serum-starved NRK cells. Fetal calf serum (FCS), PDGF, and homodimeric v-sis and PDGF A-chain protein enhanced the incorporation of 32P into pp64 over 4-fold within 30 min and over 8-fold within 2 h of exposure of NRK cells to the growth factors. In contrast, constitutive phosphorylation of 32P-labeled pp64 in nuclei of NRK cells transformed by the simian sarcoma virus (SSV) was high and only minimally stimulated by PDGF and FCS. 32P-Labeled pp64 was isolated from nuclei of PDGF-stimulated nontransformed NRK cells; the 32P of pp64 was labile in 1 M KOH, and pp64 was not significantly recognized by anti-phosphotyrosine antisera, suggesting that the PDGF-induced phosphorylation of pp64 occurred on serine or on threonine residues. However, pp64 from SSV-transformed NRK cell nuclei was significantly stable to base hydrolysis and was immunoprecipitated with anti-phosphotyrosine antisera, suggesting that pp64 from SSV-transformed cell nuclei is phosphorylated also on tyrosine. FCS, PDGF, and PDGF A- and B-chain homodimers thus stimulate the rapid time-dependent phosphorylation of a 64-kDa nuclear protein shortly after stimulation of responsive cells. The growth factor-stimulated phosphorylation of pp64 and the constitutive high levels of pp64 phosphorylation in cells transformed by SSV suggest important roles for pp64 and perhaps regulated nuclear protein kinases and phosphatases in cell division and proliferation.

  20. A highly sensitive and selective fluorescent sensor for detection of Al(3+) using a europium(III) quinolinecarboxylate.

    PubMed

    Xu, Wentao; Zhou, Youfu; Huang, Decai; Su, Mingyi; Wang, Kun; Hong, Maochun

    2014-07-07

    Eu2PQC6 has been developed to detect Al(3+) by monitoring the quenching of the europium-based emission, with the lowest detection limit of ∼32 pM and the quantitative detection range to 150 μM. Eu2PQC6 is the first ever example that the europium(III) complex serves as an Al(3+) fluorescent sensor based on "competition-displacement" mode.

  1. Calcium and protein phosphorylation in the transduction of gravity signal in corn roots

    NASA Technical Reports Server (NTRS)

    Friedmann, M.; Poovaiah, B. W.

    1991-01-01

    The involvement of calcium and protein phosphorylation in the transduction of gravity signal was studied using corn roots of a light-insensitive variety (Zea mays L., cv. Patriot). The gravitropic response was calcium-dependent. Horizontal placement of roots preloaded with 32P for three minutes resulted in changes in protein phosphorylation of polypeptides of 32 and 35 kD. Calcium depletion resulted in decreased phosphorylation of these phosphoproteins and replenishment of calcium restored the phosphorylation.

  2. Apollo 11 Cmdr Neil Armstrong watches STS-83 launch

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Apollo 11 Commander Neil A. Armstrong and his wife, Carol, were among the many special NASA STS-83 launch guests who witnessed the liftoff of the Space Shuttle Columbia April 4 at the Banana Creek VIP Viewing Site at KSC. Columbia took off from Launch Pad 39A at 2:20:32 p.m. EST to begin the 16-day Microgravity Science Laboratory-1 (MSL-1) mission.

  3. Characterization of the major phosphoprotein and its kinase on the surface of the rat adipocyte

    SciTech Connect

    Kang, E.S.; Chiang, T.M.

    1986-12-01

    Intact rat fat cell exposed to 12.5 ..mu..M (..gamma..-32P)ATP incorporate label into specific proteins within minutes. By solubilizing the reaction mixture with SDS which bypasses the subcellular fractionation steps, the labeled proteins can be identified in autoradiographs of SDS-PAGE gels. The most prominently labeled protein has an M/sub r/ of 42,000. Localization of this component to the cell surface can be made on the basis of inhibition of phosphorylation by addition of a protein derived from the rat brain with protein kinase inhibitory property, susceptibility of the phosphorylated protein to the tryptic digestion, inhibition of phosphorylation of this protein after brief exposure to melittin. To rule out the possibility that the cell surface protein might be a mitochondrial contaminant from broken cells, /sup 32/Pi-labeled and (..gamma..-/sup 32/P)ATP-labeled cells were chromatographed on a rabbit anti-pyruvate dehydrogenase antibody-Sepharose 4B column. A single labeled peak was detected upon elution of the bound fraction only in the /sup 32/pi-labeled sample, and not in the (..gamma..-/sup 32/P)ATP-labeled sample. Subcellular fractionation studies of intact cells labeled depending on whether a continuous sucrose gradient or a discontinuous sucrose gradient was used. Finally, comparison of the autoradiographs of two-dimensional (2D) gels show different isoelectric points for 42,000 M/sub r/ components in (..gamma..-/sup 32/P)ATP- and /sup 32/Pi-labeled cells. These and other experiments support the likelihood that phosphoproteins of 42,000 M/sub r/ are present at two sites in the intact rat fat cell, the cell surface and at an intracellular site, most likely the mitochondria.

  4. Apollo 11 Cmdr Neil Armstrong watches STS-83 launch

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Apollo 11 Commander Neil A. Armstrong and his wife, Carol, were among the many special NASA STS-83 launch guests who witnessed the liftoff of the Space Shuttle Columbia April 4 at the Banana Creek VIP Viewing Site at KSC. Columbia took off from Launch Pad 39A at 2:20:32 p.m. EST to begin the 16-day Microgravity Science Laboratory-1 (MSL-1) mission.

  5. Metabolic labeling of leucine rich repeat kinases 1 and 2 with radioactive phosphate.

    PubMed

    Taymans, Jean-Marc; Gao, Fangye; Baekelandt, Veerle

    2013-09-18

    Leucine rich repeat kinases 1 and 2 (LRRK1 and LRRK2) are paralogs which share a similar domain organization, including a serine-threonine kinase domain, a Ras of complex proteins domain (ROC), a C-terminal of ROC domain (COR), and leucine-rich and ankyrin-like repeats at the N-terminus. The precise cellular roles of LRRK1 and LRRK2 have yet to be elucidated, however LRRK1 has been implicated in tyrosine kinase receptor signaling, while LRRK2 is implicated in the pathogenesis of Parkinson's disease. In this report, we present a protocol to label the LRRK1 and LRRK2 proteins in cells with (32)P orthophosphate, thereby providing a means to measure the overall phosphorylation levels of these 2 proteins in cells. In brief, affinity tagged LRRK proteins are expressed in HEK293T cells which are exposed to medium containing (32)P-orthophosphate. The (32)P-orthophosphate is assimilated by the cells after only a few hours of incubation and all molecules in the cell containing phosphates are thereby radioactively labeled. Via the affinity tag (3xflag) the LRRK proteins are isolated from other cellular components by immunoprecipitation. Immunoprecipitates are then separated via SDS-PAGE, blotted to PVDF membranes and analysis of the incorporated phosphates is performed by autoradiography ((32)P signal) and western detection (protein signal) of the proteins on the blots. The protocol can readily be adapted to monitor phosphorylation of any other protein that can be expressed in cells and isolated by immunoprecipitation.

  6. DNA probe for lactobacillus delbrueckii

    SciTech Connect

    Delley, M.; Mollet, B.; Hottinger, H. )

    1990-06-01

    From a genomic DNA library of Lactobacillus delbrueckii subsp. bulgaricus, a clone was isolated which complements a leucine auxotrophy of an Escherichia coli strain (GE891). Subsequent analysis of the clone indicated that it could serve as a specific DNA probe. Dot-blot hybridizations with over 40 different Lactobacillus strains showed that this clone specifically recognized L. delbrueckii subsp. delbrueckii, bulgaricus, and lactis. The sensitivity of the method was tested by using an {alpha}-{sup 32}P-labeled probe.

  7. Apollo 7 Cmdr Wally Schirra watches STS-83 launch

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Apollo 7 Commander Walter M. 'Wally' Schirra and his wife, Josephine, were among the many special NASA STS-83 launch guests who witnessed the liftoff of the Space Shuttle Columbia April 4 at the Banana Creek VIP Viewing Site at KSC. Columbia took off from Launch Pad 39A at 2:20:32 p.m. EST to begin the 16-day Microgravity Science Laboratory-1 (MSL-1).

  8. Calcium and protein phosphorylation in the transduction of gravity signal in corn roots

    NASA Technical Reports Server (NTRS)

    Friedmann, M.; Poovaiah, B. W.

    1991-01-01

    The involvement of calcium and protein phosphorylation in the transduction of gravity signal was studied using corn roots of a light-insensitive variety (Zea mays L., cv. Patriot). The gravitropic response was calcium-dependent. Horizontal placement of roots preloaded with 32P for three minutes resulted in changes in protein phosphorylation of polypeptides of 32 and 35 kD. Calcium depletion resulted in decreased phosphorylation of these phosphoproteins and replenishment of calcium restored the phosphorylation.

  9. X-Ray Laser

    DTIC Science & Technology

    1991-01-31

    32. p. 4358. 1985. [11] R. A. Fisher. Appi. PhYs. Lett.. vol. 24. p. 598. 1974. 141 R.- K. Klein et al.. Phys. Rev. B, vol. 3 1. p. 68 . 1985. [121 M...S. Datz. J. 0. Kephart. R. K. Klein. R. H. Pan- ’D. Marcuse . Lizht Transmission Optics Van Nostrand- t and H. Park, Nucl. Instrum. Methods B 10/11

  10. Sensitivity to disgust and perceptions of natural bodies of water and watercraft activities.

    PubMed

    Bixler, Robert D; Powell, Gwynn

    2003-08-01

    A written 7-item self-report scale on sensitivity to disgust and participation in watercraft activities was administered to 450 seasonal park employees. Correlations indicate that nonparticipation in seven different watercraft sports was weakly related with reactions of disgust to contact with natural bodies of water (r(pbis) = -.12 to -.32, p < .01). Negative affective reactions to contact with natural bodies of water may contribute to panic reactions in watercraft accidents.

  11. Primary structure of tRNA-Lys of E. coli B.

    PubMed Central

    Chakraburtty, K; Steinschneider, A; Case, R V; Mehler, A H

    1975-01-01

    The primary structure of tRNALys of E. coli was determined by use of [32P]-tRNA. The sequence is pGGGUCGUUAGCUCAGDDGGDAGAGCAGUUGACUmam5-s2-UUU-t6AApsiCAAUUGm7GXCGCAGGTpsiCGAAUCCUGCACGACCCACCA. No s4-U was detected in position 8. No other lysine tRNA was detected but the existence of another species has not been ruled out. Images PMID:802509

  12. An Applied Overview of Moving Boundary Problems.

    DTIC Science & Technology

    1977-08-01

    be exhibited by the austenitic steel in [3.2], but not by the delta - ferritic iron of (3.3], the latter however lending itself to a simple relation...Series, Plenum Publ. Corp., New York. 3.2 P.J. Wray, “Plastic Deformations of Delta - Ferritic Iron at Intermediate Strain Rates,” Metell. Trans A , vol. 7A...Art and Science of Growing Cr ystals , 1. Wiley I!)’~ Sons , ~ew York (1963). _ _- V

  13. Synthesis, Properties and Applications of Diazotrifluropropanoyl-Containing Photoactive Analogues of Farnesyl Diphosphate Containing Modified Linkages for Enhanced Stability

    PubMed Central

    Hovlid, Marisa L.; Edelstein, Rebecca L.; Henry, Olivier; Ochocki, Joshua; DeGraw, Amanda; Lenevich, Stepan; Talbot, Trista; Young, Victor G.; Hruza, Alan W.; Lopez-Gallego, Fernando; Labello, Nicholas P.; Strickland, Corey L.; Schmidt-Dannert, Claudia; Distefano, Mark D.

    2009-01-01

    Photoactive analogues of farnesyl diphosphate (FPP) are useful probes in studies of enzymes that employ this molecule as a substrate. Here, we describe the preparation and properties of two new FPP analogues that contain diazotrifluoropropionyl photophores linked to geranyl diphosphate via amide or ester linkages. The amide-linked analogue (3) was synthesized in 32P-labeled form from geraniol in 7 steps. Experiments with Saccharomyces cerevisiae protein farnesyltransferase (ScPFTase) showed that 3 is an alternative substrate for the enzyme. Photolysis experiments with [32P]3 demonstrate that this compound labels the β-subunits of both farnesyl- and geranylgeranyltransferase (types 1 and 2). However, the amide-linked probe 3 undergoes a rearrangement to a photochemically unreactive isomeric triazolone upon long term storage making it inconvenient to use. To address this stability issue, the ester-linked analogue 4 was prepared in 6 steps from geraniol. Computational analysis and X-ray crystallographic studies suggest that 4 binds to PFTase in a similar fashion as FPP. Compound 4 is also an alternative substrate for PFTase and a 32P-labeled form selectively photocrosslinks the β-subunit of ScPFTase as well as E. coli farnesyldiphosphate synthase and a germacrene-producing sesquiterpene synthase from Nostoc sp. strain PCC7120 (a cyanobacterial source). Finally, nearly exclusive labeling of ScPFTase in crude E. coli extract was observed, suggesting that [32P]4 manifests significant selectivity and should hence be useful for identifying novel FPP utilizing enzymes in crude protein preparations. PMID:19954434

  14. Genotoxicity of the fungicide dichlofluanide in seven assays

    SciTech Connect

    Heil, J.; Reifferscheid, G.; Hellmich, D.; Hergenroeder, M.; Zahn, R.K. )

    1991-01-01

    Seven different endpoints for detection of genotoxicity have been used to demonstrate the DNA-altering properties of Dichlofluanid, a fungicide commonly used in viticulture pest control. Each endpoint (DNA synthesis inhibition test, alkaline viscosimetry, umu-test, alkaline filter elution, FADU-test, {sup 32}P-postlabeling, and electron microscopy) shows clear evidence of genotoxicity. These data indicate that application of the fungicide dichlofluanid may be mutagenic and/or carcinogenic for exposed humans.

  15. Genetic and antigenic relatedness of bovine herpesvirus-1 and pseudorabies virus

    SciTech Connect

    Bush, C.E.

    1985-01-01

    The DNA sequence homology between the genomes of bovine herpesvirus-1 (BHV-1) and pseudorabies virus (PRV) was examined. Reciprocal cross hybridization of viral DNA labeled by nick translation to Southern blots of Kpnl, BamH1, EcoR1, and HindIII restriction endonuclease digested DNA, detected homologous sequences dispersed throughout the genomes of the two viruses. The DNA-DNA hybrids were found to be stable under high stringency wash conditions. Sequences of a /sup 32/P-labeled PRV DNA A fragment probe were found to hybridize only to the BHV-1 HindIII G fragment. This indicated that the sequence homology detected between these two viruses was not simply due to fortuitous hybridization of guanine plus cytosine rich sequences. The homology between BHV-1 and PRV was determined by liquid reassociation. It was found that the hybridization rates between /sup 32/P-labeled PRV DNA and unlabeled BHV-1 DNA and /sup 32/P-labeled BHV-1 DNA and unlabeled PRV DNA corresponded to approximately 8% reassociation. The antigenic relatedness between BHV-1 and PRV was also examined. Eighty percent plaque reduction serum neutralization tests showed that BHV-1 rabbit hyperimmune antiserum neutralized BHV-1 virus at a serum neutralization titer (SNT) of 1:256 and PRV virus at an (SNT) of 1:8. PRV rabbit hyperimmune antiserum neutralized PRV virus at an SNT of 1:4 and BHV-1 virus at an SNT of 1:2.

  16. Phosphorylation of partially purified 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine:acetyl-CoA acetyltransferase from rat spleen.

    PubMed Central

    Gomez-Cambronero, J; Mato, J M; Vivanco, F; Sanchez-Crespo, M

    1987-01-01

    A new improved method for purification of the enzyme 1-O-alkyl-2-lyso-sn-glycero-3-phosphocholine: acetyl-CoA acetyltransferase (EC 2.3.1.67) from rat spleen is described. The catalytic subunit of cyclic AMP-dependent protein kinase in the presence of MgATP stimulated about 3-fold the activity of this partially purified enzyme activity. When [gamma-32P]ATP was included in the assay mixture, the analysis of phosphoprotein products by SDS/polyacrylamide-gel electrophoresis and autoradiography showed the incorporation of [32P]phosphate into a single protein band of about 30 kDa. Analysis of the phosphorylated amino acids indicated that the phosphate was incorporated into a serine residue. Activation of the acetylation reaction by the protein kinase was reversible. The reversal of the activation was coincident with the loss of the [32P]phosphate incorporated into the 30 kDa protein band, which suggests that the acetyltransferase is regulated by a phosphorylation-dephosphorylation mechanism dependent on cyclic AMP. Images Fig. 2. Fig. 3. Fig. 4. PMID:3663199

  17. Pharmaceutical issues during the review of European Marketing Authorisation Applications in Malta().

    PubMed

    Chetcuti, Michael; Serracino-Inglott, Anthony; Flores, Gavril; Borg, John J

    2017-08-07

    The aim of this study was to identify pharmaceutical issues encountered during regulatory review in European Procedures. A database of issues from Day 70 assessment reports of 150 EU procedures was compiled; most procedures were for generics (108). Frequencies of common deficiencies have been calculated and summarised for use of all stakeholders. Out of the 150 procedures reviewed, covering 309 products, a total of 4796 concerns were identified. Of these concerns, 167 were Potential Serious Risks to Public Health, 67 were raised on drug substance and 100 on the drug product. The distribution of total concerns was as follows: 2168 concerns on drug substance and 2584 on drug product. Most concerns raised were on control of drug substance and drug product (834 & 626 for 3.2.S.4 and 3.2.P.5, respectively), followed by concerns on the manufacturing (482 & 564 for 3.2.S.2 and 3.2.P.3, respectively) and stability 147 & 398 for 3.2.S.7 and 3.2.P.8, respectively). In conclusion, the frequencies and trends of identified deficiencies together with their impact were discussed from a regulatory point of view. The main findings indicate that applicants would benefit from following published guidelines so that delays in the registration of medicines could be avoided.

  18. Radioprotection against lethal damage caused by chronic irradiation with radionuclides in vitro.

    PubMed

    Howell, R W; Goddu, S M; Bishayee, A; Rao, D V

    1998-10-01

    To examine the capacity of chemical protectors to mitigate damage caused by chronic irradiation by incorporated radionuclides in vitro, cells must be maintained in the presence of the protector during the course of the irradiation. Such long exposures to chemical protectors at concentrations high enough to afford protection usually results in extreme chemotoxicity. To overcome this problem, experimental conditions were developed to allow Chinese hamster V79 cells to be maintained in 5% DMSO for prolonged periods (up to 72 h) with no observable chemotoxicity. Under these conditions, the capacity of DMSO to protect against damage to V79 cells caused by unbound 32P and 3H2O and DNA-incorporated (131)IdU, [3H]dThd and 125IdU was examined. The dose modification factors for 32P, 3H2O, (131)IdU, [3H]dThd and 125IdU were 2.6+/-0.5, 2.3+/-0.3, 1.0+/-0.1, 1.16+/-0.07 and 1.07+/-0.02, respectively. These results show that 5% DMSO is capable of protecting cultured V79 cells against lethal damage caused by beta particles emitted by unbound 32P and 3H2O, whereas little or no protection is afforded against damage caused by beta particles emitted by DNA-incorporated (131)I and 3H or low-energy Auger electrons emitted by DNA-incorporated 125I.

  19. Possible role of firmly bound ATP in the energy transduction of photosynthetic membranes.

    PubMed

    Lutz, H U; Beyeler, W; Pflugshaupt, C; Bachofen, R

    1975-01-01

    Chromatophores of Rhodospirillum rubrum and spinach chloroplasts contain firmly bound ATP that is rapidly labeled along with ADP in the presence of 32Pi and endogenous nucleotides. The labeling is not entirely dependent on light. In chloroplasts three types of bound ATP can be defined methodologically by their extraction properties: buffer-soluble; acid-soluble; and SDS-soluble or firmly bound ATP. Extensive washing of the chloroplasts does reduce buffer-soluble but not acid-soluble and firmly bound ATP. Buffer-soluble [32P] ATP is almost exclusively gamma labeled while acid-soluble and firmly bound ATP are labeled in the beta and gamma position equally. CCCP, desaspidin, and phlorizin do not inhibit the labeling of firmly bound ATP, whereas the phosphorylation is almost abolished. However, EDTA and NEM pretreatments of the chloroplasts affect both reactions similarly. The postillumination [32P] ATP synthesis with chromatophores can be inhibited by adding ATP to the incubation mixture after illumination if 32Pi is included only during the dark incubation, but is without effect if 32Pi is present only during illumination. On the other hand, ADP added after illumination inhibits post-illumination [32P] ATP formation in both chromatophores and chloroplasts only if 32Pi is present during illumination. The data can be explained by a coupling factor having two sites, as proposed previously on the basis that firmly bound ATP does not transfer its phosphoryl group but seems to drive a synthesis of acid-soluble ATP which incorporates free phosphate.

  20. Inositol 1,4,5-trisphosphate binds to a specific receptor and releases microsomal calcium in the arterior pituitary gland

    SciTech Connect

    Guillemette, G.; Balla, T.; Baukal, A.J.; Catt, K.J.

    1987-12-01

    The properties of inositol 1,4,5-trisphosphate (InsP/sub 3/) receptor sites in the anterior pituitary were evaluated by binding studies with InsP/sub 3/ labeled with /sup 32/P to high specific radioactivity. Specific binding of Ins(/sup 32/P)P/sub 3/ was demonstrable in pituitary membrane preparations and was linearly proportional to the amount of membrane added over the range 0.5-2 mg of protein. Kinetic studies showed that specific InsP/sub 3/ binding was half-maximal in about 40 sec and reached a plateau after 15 min at 0/sup 0/C. Scatchard analysis of the binding data was consistent with a single set of high affinity sites. The specificity of Ins(/sup 32/P)P/sub 3/ binding to these sites was illustrated by the much weaker affinity for structural analogs such as inositol 1-phosphate, phytic acid, 2,3-bisphosphoglycerate, and fructose 1,6-bisphosphate. To assess the functional relevance of the InsP/sub 3/ binding sites, the Ca/sup 2 +/-releasing activity of InsP/sub 3/ was measured in pituitary membrane preparations. Under physiological conditions within the cytosol, the high-affinity InsP/sub 3/ binding sites characterized in pituitary membranes could serve as the putative receptors through which InsP/sub 3/ triggers Ca/sup 2 +/ mobilization in the anterior pituitary gland.

  1. Combinatorial control of diverse metabolic and physiological functions by transcriptional regulators of the yeast sulfur assimilation pathway

    PubMed Central

    Petti, Allegra A.; McIsaac, R. Scott; Ho-Shing, Olivia; Bussemaker, Harmen J.; Botstein, David

    2012-01-01

    Methionine abundance affects diverse cellular functions, including cell division, redox homeostasis, survival under starvation, and oxidative stress response. Regulation of the methionine biosynthetic pathway involves three DNA-binding proteins—Met31p, Met32p, and Cbf1p. We hypothesized that there exists a “division of labor” among these proteins that facilitates coordination of methionine biosynthesis with diverse biological processes. To explore combinatorial control in this regulatory circuit, we deleted CBF1, MET31, and MET32 individually and in combination in a strain lacking methionine synthase. We followed genome-wide gene expression as these strains were starved for methionine. Using a combination of bioinformatic methods, we found that these regulators control genes involved in biological processes downstream of sulfur assimilation; many of these processes had not previously been documented as methionine dependent. We also found that the different factors have overlapping but distinct functions. In particular, Met31p and Met32p are important in regulating methionine metabolism, whereas p functions as a “generalist” transcription factor that is not specific to methionine metabolism. In addition, Met31p and Met32p appear to regulate iron–sulfur cluster biogenesis through direct and indirect mechanisms and have distinguishable target specificities. Finally, CBF1 deletion sometimes has the opposite effect on gene expression from MET31 and MET32 deletion. PMID:22696679

  2. Methods to distinguish various types of protein phosphatase activity

    SciTech Connect

    Brautigan, D.L.; Shriner, C.L.

    1988-01-01

    To distinguish the action of protein Tyr(P) and protein Ser(P)/Thr(P) phosphatases on /sup 32/P-labeled phosphoproteins in subcellular fractions different inhibitors and activators are utilized. Comparison of the effects of added compounds provides a convenient, indirect method to characterize dephosphorylation reactions. Protein Tyr(P) phosphatases are specifically inhibited by micromolar Zn2+ or vanadate, and show maximal activity in the presence of EDTA. The other class of cellular phosphatases, specific for protein Ser(P) and Thr(P) residues, are inhibited by fluoride and EDTA. In this class of enzymes two major functional types can be distinguished: those sensitive to inhibition by the heat-stable protein inhibitor-2 and not stimulated by polycations, and those not sensitive to inhibition and stimulated by polycations. Preparation of /sup 32/P-labeled Tyr(P) and Ser(P) phosphoproteins also is presented for the direct measurement of phosphatase activities in preparations by the release of acid-soluble (/sup 32/P)phosphate.

  3. Phosphorylation of proteins in Dictyostelium discoideum during development

    SciTech Connect

    Coffman, D.S.

    1982-01-01

    The phosphoproteins in D. discoideum were studied with respect to their formation, metabolic stability, cellular and subcellular distribution. Special emphasis was on the role of cAMP on the pattern of phosphorylation. Amoebae were metabolically labeled with /sup 32/P/sub i/; subsequently proteins of the total lysate, nuclei and membranes were resolved by SDS-polyacrylamide gel electrophoresis and subjected to autoradiography. Numerous changes in the profile of phosphoproteins were observed during development. Functions were assigned to four membranal phosphoproteins; only one protein, the heavy chain of myosin, was susceptible to phosphorylation in vitro when purified membranes and /sup 32/P-ATP were used. A comparison between the time of protein synthesis and phosphorylation, as examined in vivo using /sup 35/S-methionine and /sup 32/P/sub i/ labeling of amoebae and two-dimensional gel electrophoresis, indicated that phosphorylation is concurrent with synthesis. It appears then that there are two classes of membranal phosphoproteins in D. discoideum which differ with respect to the stability of the phosphate moiety. It is evident that the turnover of the phosphate moiety in myosin heavy chain plays a crucial role in the function of myosin; a role for the metabolically inert phosphate of other membranal proteins remains to be established. The G protein which couples occupancy of hormone receptor to stimulation of adenylate cyclase in higher multicellular eukaryotes was detected in D. discoideum. The G protein is present in approximately equal amounts in vegetative and in developing amoebae.

  4. Sexual maturation and productivity of Japanese quail fed graded concentrations of mercuric chloride.

    PubMed

    Hill, E F; Shaffner, C S

    1976-07-01

    Japanese quail (Coturnix c. japonica) were fed 0, 2, 4, 8, 16, and 32 p.p.m. Hg as mercuric chloride (HgCl2) from the time of hatching up to the age of 1 year. None of the birds manifested any gross signs of mercury poisioning. Food consumption, growth rate, and weight maintenance were unaffected. Initial oviposition tended to occur at a younger age as dietary mercuric chloride increased, e.g., the median age at which egg laying began among hens fed 32 p.p.m. Hg was 6 days younger than for controls. The average rate of egg production was positively related to the concentration of mercuric chloride with the most pronounced differences between treatments occurring among young (less than 9-week-old) hens. Beyond 9 weeks of age production was more uniform among the treatments, but even after 1 year hens on 32 p.p.m. Hg were laying an average of 13.5% more eggs than controls. Rate of egg fertilization was generally depressed for all Hg-treatments above 4 p.p.m. Hatchability of fertilized eggs and eggshell thickness appeared unaffected by mercuric chloride.

  5. Chromosomal DNA probes for the identification of asaccharolytic anaerobic pigmented bacterial rods from the oral cavity of cats.

    PubMed

    Love, D N; Bailey, G D; Bastin, D

    1992-06-01

    A dot-blot hybridisation assay using isolated high molecular weight DNA as whole chromosomal probes of the cat pigmented asaccharolytic Bacteroides/Porphyromonas species was used against both purified high molecular weight DNA and DNA released on membranes from whole cells for the identification of B. salivosus and for its differentiation from the other anaerobic species isolated from normal and diseased mouths of cats and horses. 32P-labelled probes were compared with digoxigenin (DIG)-labelled probes (Boehringer-Mannheim). The whole chromosomal probes were specific--differentiating B. salivosus from a variety of species (including members of the genera Bacteroides, Fusobacterium, Eubacterium, and Prevotella) found in normal and abnormal mouths of cats and horses. Likewise, asaccharolytic black pigmented Group 2 strains were distinguishable from all strains tested. However, cat strains of P. gingivalis which show 68-76% DNA-DNA homology with human strain P. gingivalis ATCC 33277T, were not distinguishable from each other using either 32P-labelled or DIG-labelled probes. The minimum amount of pure Bacteroides DNA which could be detected by the 32P-labelled probe was 100-300 pg, while the amount of pure DNA detected by the DIG system was 1-3 mg after room temperature colour development for 1 h and 100-300 pg after 6 h colour development.

  6. Multiple structures of adeno-associated virus DNA: analysis of terminally labeled molecules with endonuclease R-Hae III.

    PubMed

    Denhardt, D T; Eisenberg, S; Bartok, K; Carter, B J

    1976-05-01

    The double-stranded form of adeno-associated virus (AAV) DNA has about 20 sites sensitive to endonuclease R.Hae III from Haemophilus aegypitus; the fragments produced fall into about 13 size classes, 8 of which contain single fragments. The location of the Hae III-produced AAV fragments relative to the three EcoR1 fragments was determined. Using revised figures for the molecular weights of the Hae III cleavage products of phiX174 replicative form DNA, we calculated that AAV DNA contains about 4,000 nucleotides. After Hae III digestiion of duplex DNA terminally labeled with 32P using polynucleotide kinase, the majority of fragments containing a 5' 32P label were about 40 nucleotides in length, and fragments of similar size were generated from each end, suggesting that the Hae site closest to the end is within the terminal repetition. Two more-slowly-migrating cleavage products also bore 5' 32P end label. These three terminally labeled species were also generated from single-stranded AAV DNA by digestion with Hae III, and evidence that one may have a nonlinear ("rabbit-ear") structure is presented. The predominant 5' terminal base was identified as thymine for both the plus and minus strands of AAV. Single-stranded AAV molecules could not be efficiently covalently circularized by incubation with polynucleotide ligase or ligase plus T4 DNA polymerase.

  7. Comparative effect of microwaves and boiling on the denaturation of DNA

    SciTech Connect

    Stroop, W.G.; Schaefer, D.C. )

    1989-11-01

    The effect of heat and microwave denaturation of small volumes of double-stranded plasmid DNA has been compared. Samples of intact plasmid DNA had plasmid DNA linearized by digestion with EcoRI were conventionally denatured in a boiling water bath or denatured by 2450 MHz of microwave energy for 0-300 s. Heat denaturation for periods longer than 120 s caused breakdown of linearized plasmid DNA; however, microwave denaturation for 10-300 s caused no apparent degradation of linearized DNA. Breakdown of DNA forms II and III was noted in plasmid DNA subjected to 300 s of either heat or microwave denaturation but breakdown of forms II and III occurred more quickly with heat than with microwave treatment. Microwave treatment was also found to be better than heat to denature 32P-labeled DNA probes subsequently used to detect homologous DNA samples immobilized on nitrocellulose filters. A microwave-treated 32P-labeled DNA probe was able to hybridize to DNA samples 20 times more dilute than a heat-treated 32P-labeled DNA probe. Depending on the form of DNA to be analyzed, these results indicate that small volumes of DNA solutions and radiolabeled DNA probes can be effectively denatured in a conventional microwave oven.

  8. Evidence for the presence and role of tightly bound adenine nucleotides in phospholipid-free purified Micrococcus lysodeikticus adenosine triphosphatase.

    PubMed

    Muñoz, C; Palacios, P; Muñoz, E

    1977-10-01

    [32P]-labeled ATPase was isolated in a highly purified state from Micrococcus lysodeikticus strain PNB grown in medium supplemented with [32P]orthophosphate. Selective extraction procedures allowed us to determine that at least 25% of the firmly bound label belonged to adenine nucleotides, ATP and ADP being present in equimolar amounts. However, no 32P label was found to be part of phospholipids. This was confirmed by purification of the ATPase from cells fed with [2-3H]glycerol. Using the luciferin-luciferase assay we estimated that ATPase freshly isolated by Sephadex chromatography (specific activity 10-14 micromole substrate transformed x min(-1) x mg protein(-1)) contained 2 moles ATP/mole of enzyme. The ratio fell with the age of enzyme and its purification by gel electrophoresis and this was paralleled by a loss of ATPase activity. The endogenous nucleotides were readily exchanged by added ADP or ATP. This result suggests that the sites for tight binding of adenine nucleotides are equivalent, although ADP seems to have a higher affinity for them. The last properties represent a peculiar characteristic of this bacterial ATPase as compared with other bacterial and organelle energy-transducing proteins.

  9. State of hepatitis B viral DNA in a human hepatoma cell line.

    PubMed Central

    Marion, P L; Salazar, F H; Alexander, J J; Robinson, W S

    1980-01-01

    PLC/PRF/5, a tissue culture cell line isolated from a human hepatocellular carcinoma and producing hepatitis B surface antigen, was studied for the presence of hepatitis B virus (HBV)-specific DNA and RNA. PLC/PRF/5 cell DNA accelerated the rate of reassociation of HBV [32P]DNA, and quantitative experiments indicated that the cells contained approximately four copies of viral DNA per haploid, mammalian cell DNA equivalent. PLC/PRF/5 DNA accelerated the rate of reassociation of all individual restriction endonucleases HincII and HaeIII fragments of HBV [32P]DNA, indicating that DNA from all regions of the viral genome is present in the cells. This suggests that these cells contain at least most, and possibly all, of the viral genome. Digestion of PLC/PRF/5 cell DNA with restriction endonuclease HindIII (an enzyme found not to cleave the DNA of any HBV isolate so far examined) yielded only three fragments, all larger than virion DNA, which contained HBV DNA base sequences, suggesting that HBV DNA is integrated in high-molecular-weight DNA at three different sites in these cells and that there is no viral DNA in an episomal form. PLC/PRF/5 cell [32P]RNA was found to hybridize with all restriction fragments of HBV DNA adequately tested, indicating that at least most, and possibly all, of the viral DNA in these cells is transcribed. Images PMID:6251250

  10. Prostaglandin F2 alpha administered in vivo induces Ca2+-dependent protein phosphorylation in rat luteal tissue

    SciTech Connect

    Baum, M.S.

    1989-01-01

    The present study was performed in order to further elucidate the mechanism of action of PGF2 alpha in luteolysis in the rat ovary. Seven days after priming with superovulatory doses of pregnant mare serum gonadotropin and human chorionic gonadotropin to induce luteal tissue formation, the rats were injected with a luteolytic dose of the prostaglandin F2 alpha analogue cloprostenol. The ovaries were then homogenized, a 30,000 x g supernatant and pellet were prepared, whereafter aliquots of the preparations were incubated in the presence of (gamma-/sup 32/P)ATP with or without Ca2+. The phosphorylated proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and localized by autoradiography. The presence of Ca2+ caused an increased phosphorylation of a 45 kDa protein band in the particulate, but not in the cytosol, fraction. Furthermore, PGF2 alpha rapidly increased the /sup 32/P incorporation into the same protein band of 45 kDa. Thus, the PGF2 alpha-stimulated /sup 32/P incorporation was Ca2+-dependent and seen only in the particulate fraction. These results suggest that PGF2 alpha in its role as a luteolytic agent stimulates a Ca2+-dependent phosphorylation of a specific protein in luteal membranes of the rat ovary.

  11. Radioactive Phosphorylation of Alcohols to Monitor Biocatalytic Diels-Alder Reactions

    PubMed Central

    Nierth, Alexander; Jäschke, Andres

    2011-01-01

    Nature has efficiently adopted phosphorylation for numerous biological key processes, spanning from cell signaling to energy storage and transmission. For the bioorganic chemist the number of possible ways to attach a single phosphate for radioactive labeling is surprisingly small. Here we describe a very simple and fast one-pot synthesis to phosphorylate an alcohol with phosphoric acid using trichloroacetonitrile as activating agent. Using this procedure, we efficiently attached the radioactive phosphorus isotope 32P to an anthracene diene, which is a substrate for the Diels-Alderase ribozyme—an RNA sequence that catalyzes the eponymous reaction. We used the 32P-substrate for the measurement of RNA-catalyzed reaction kinetics of several dye-labeled ribozyme variants for which precise optical activity determination (UV/vis, fluorescence) failed due to interference of the attached dyes. The reaction kinetics were analyzed by thin-layer chromatographic separation of the 32P-labeled reaction components and densitometric analysis of the substrate and product radioactivities, thereby allowing iterative optimization of the dye positions for future single-molecule studies. The phosphorylation strategy with trichloroacetonitrile may be applicable for labeling numerous other compounds that contain alcoholic hydroxyl groups. PMID:21731729

  12. Spontaneous hematologic recovery from bone marrow aplasia after accidental tenfold overdosage with radiophosphorus

    SciTech Connect

    Gmuer, J.; Bischof, B.; Coninx, S.; Bucher, U.; Poretti, G.; Henrichs, K.; Kaul, A.; Roedler, H.D.; Buettner, K.; Frick, P.G.

    1983-04-01

    Two patients with polycythemia vera received intravenously an accidental tenfold overdosage of radiophosphorus therapy (60 and 50 mCi 32P, respectively). In both patients, the occurrence of hemorrhagic complications 3 wk after the 32P medication led to detection of the error and referral to our hospital. Upon admission they showed an agranulocytosis, severe thrombocytopenia, and bone marrow aplasia. In both cases, spontaneous recovery of the hematopoiesis was observed from day 40 posttreatment onward. In one patient, a slow but ultimately complete normalization of blood counts and marrow morphology took place, whereas in the other, a mild thrombocytopenia persists. Nearly 5 yr after the accidental overdosage, both patients are clinically well. Symptoms of polycythemia vera have not reappeared up to now. Attempts were made to evaluate the radiation dose absorbed by the bone marrow. In the first patient, the daily 32P excretion was determined from day 22 to day 60, whereas in the other patient a whole body count was performed on day 78 after administration. From these results, an approximate cumulative bone marrow dose of 10 Sv (1000 rem) could be calculated.

  13. Isoproterenol stimulates phosphorylation of the insulin-regulatable glucose transporter in rat adipocytes.

    PubMed Central

    James, D E; Hiken, J; Lawrence, J C

    1989-01-01

    We have examined the acute effects of insulin and isoproterenol on the phosphorylation state of the insulin-regulatable glucose transporter (IRGT) in rat adipocytes. The IRGT was immunoprecipitated from either detergent-solubilized whole-cell homogenates or subcellular fractions of 32P-labeled fat cells and subjected to sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The 32P-labeled IRGT was detected by autoradiography as a species of apparent Mr 46,000. Insulin stimulated translocation of the IRGT from low-density microsomes to the plasma membrane but did not affect phosphorylation of the transporter in either fraction. Isoproterenol inhibited insulin-stimulated glucose transport by 40% but was without effect on the subcellular distribution of the transporter in either the presence or absence of insulin. Isoproterenol stimulated phosphorylation of the IRGT 2-fold. Incubating cells with dibutyryl-cAMP and 8-bromo-cAMP also stimulated phosphorylation 2-fold, and the transporter was phosphorylated in vitro when IRGT-enriched vesicles were incubated with cAMP-dependent protein kinase and [gamma-32P]ATP. These results suggest that isoproterenol stimulates phosphorylation of the IRGT via a cAMP-dependent pathway and that phosphorylation of the transporter may modulate its ability to transport glucose. Images PMID:2554313

  14. Phosphate cycling on the basic protein of Plodia interpunctella granulosis virus

    NASA Technical Reports Server (NTRS)

    Funk, C. J.; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    The presence of infected cell-specific phosphoproteins was investigated in Plodia interpunctella granulosis virus (PiGV)-infected fat body using [32P]orthophosphoric acid labeling. One infected cell-specific phosphoprotein had a mobility similar to that of the basic protein (VP12) of PiGV. Further analysis, using immunoblotting and acid-urea gel analysis of infected fat body, confirmed that this phosphoprotein was VP12. However we did not detect phosphorylated VP12 in 32P-labeled nucleocapsids. Phosphoamino acid analysis of 32P-labeled VP12 revealed that phosphoserine was present in the basic protein. Since VP12 is phosphorylated in the infected cell, but not in the nucleocapsid, it appears that dephosphorylation of VP12 is a critical event in the life cycle of the virus. We therefore assayed virus nucleocapsids and infected fat body for the presence of phosphatase activity. Phosphatase activity was not detected in the virus, but the infected fat body had more activity than uninfected fat body. A model for nucleocapsid assembly and uncoating is presented which takes into account the phosphorylation state of VP12, the role of Zn2+ in the nucleocapsid, and the role of the capsid-associated kinase.

  15. Composition and properties of a cell-membrane fraction from the polymorphonuclear leucocyte

    PubMed Central

    Woodin, A. M.; Wieneke, Antonnette A.

    1966-01-01

    1. The composition of a vesicular cell-membrane fraction from leucocytes has been studied. The bulk of the mass is accounted for as protein and lipid. A small amount of carbohydrate, including some N-acetylneuraminic acid, is present. The phospholipid/cholesterol molar ratio is 1·4 and differs from that for the whole cell. 2. Labile phosphorus groups are present in the membrane but the analysis is complicated by the presence of phosphorus occluded in the membrane vesicles. 3. Leucocidin does not change the gross composition of the membranes or alter the amount or reactivity of the phosphorus compounds. 4. The cell-membrane fraction has considerable avidity for an impurity present in commercial [32P]orthophosphate. When this is removed [32P]orthophosphate or [32P]ATP does not label the membrane. 5. The presence of an NADH2–cytochrome c oxidoreductase and an alkaline phosphatase is described. The adenosine-triphosphatase activity of the membrane has not been found to depend on the presence of Na+ or K+. PMID:4287773

  16. A Newborn with Genital Ambiguity, 45,X/46,XY Mosaicism, a Jumping Chromosome Y, and Congenital Adrenal Hyperplasia.

    PubMed

    Zhang, Lei; Cooley, Linda D; Chandratre, Sonal R; Ahmed, Atif; Jacobson, Jill D

    2013-01-01

    Disorders of sex development (DSD), formerly termed "intersex" conditions, arise from numerous causes. CAH secondary to 21-hydroxylase deficiency is the most common cause of DSD. Sex chromosome disorders, including sex chromosome mosaicism, are the second most common cause of DSD. We discuss a medically complex neonate with DSD presenting with ambiguous genitalia. Hormone levels suggested 21-hydroxylase deficiency. Molecular analysis revealed compound heterozygous mutations in the 21-hydroxylase gene (CYP21A2), confirming the diagnosis of CAH. Chromosome analysis revealed sex chromosome mosaicism with three cell lines: 45,X[8]/45,X,tas(Y;16)(p11.32;p13.3)[8]/45,X,t(Y;8)(p11.32;p23.3)[4] with the Y chromosome in telomere association with chromosomes 8p and 16p in different cell lines, a "jumping translocation." Histologically, the right gonad had irregular, distended seminiferous tubules with hyperplastic germ cells contiguous with ovarian stroma and primordial follicles. The left gonad had scant ovarian stroma and embryonic remnants. Chromosome analyses showed mosaicism in both gonads: 45,X[17]/45,X,tas(Y;8)(p11.32;p23.3)[3]. This is the first case of coexisting CAH and 45,X/46,XY mosaicism reported in the English literature and the third case of a constitutional chromosome Y "jumping translocation." Our report documents the medical and genetic complexity of children such as this one with ambiguous genitalia and discusses the need for a multidisciplinary team approach.

  17. Phosphoinositide phosphorylation and hydrolysis in pancreatic islet cell membrane

    SciTech Connect

    Dunlop, M.E.; Malaisse, W.J.

    1986-02-01

    Membranes were isolated from dispersed rat pancreatic islet cells by attachment to Sephadex beads. When these membranes were exposed to (gamma-32P)ATP, formation of 32P-labeled phosphatidate, phosphatidylinositol 4-phosphate, and phosphatidylinositol 4,5-bisphosphate was observed. Carbamylcholine, added 10 s prior to lipid extraction, caused a dose-related fall in 32P-labeled phospholipids. The effect of the cholinergic agent was suppressed by atropine, ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid, and verapamil, and simulated, in part, by an increase in Ca2+ concentration. When the membranes were derived from islet cells prelabeled with (U-14C)arachidonate, carbamylcholine stimulation, in addition to decreasing labeled polyphosphoinositides, was accompanied by an increased production of labeled diacylglycerol, without a concomitant increase in labeled phosphatidylinositol. These results indicate that activation of a plasma membrane-associated phospholipase C directed against polyphosphoinositides represents a primary event in the functional response of the pancreatic beta cell to cholinergic agents.

  18. Formation of sugar phosphates in colicin K-treated Escherichia coli.

    PubMed Central

    Takagaki, Y; Matsuhashi, M; Yamashita, J; Horio, T

    1977-01-01

    Colicin K greatly decreased the incorporation of 32P-labeled inorganic orthophosphate into nucleotides and nucleic acids, causing a concomitant increase in the formation of 32P-labeled sugar phosphates in sensitive cells of Escherichia coli. These sugar phosphates were formed in aerobically growing cells, as well as in cells under stringent control of ribonucleic acid synthesis. The main 32P-labeled product was identified as sedoheptulose 7-phosphate in two strains (B1 and K-12 MK-1) and fructose 1,6-diphosphate in one strain (K-12 CP78). The formation of sugar phosphates induced by colicin K was inhibited by carbonyl cyanide m-chlorophenylhydrazone. It was also not observed in N,N'-dicyclohexylcarbodiimide-treated cells or Mg2+-(Ca2+)-adenosine triphosphatase-less mutant (strain K-12 AN120) cells. Thus, the formation of sugar phosphates in colicin K-treated cells is dependent on the formation of adenosine 5'-triphosphate by oxidative phosphorylation. Images PMID:141446

  19. Collagen-induced binding to human platelets of platelet-derived growth factor leading to inhibition of P43 and P20 phosphorylation

    SciTech Connect

    Bryckaert, M.C.; Rendu, F.; Tobelem, G.; Wasteson, A.

    1989-03-15

    Platelet-derived growth factor (PDGF) is known to inhibit collagen-induced platelet aggregation. Collagen-induced binding of /sup 125/I-PDGF to human washed platelets was therefore investigated. It was found to be time-dependent, reaching a plateau at 20 degrees C after 30 min, collagen concentration-dependent, specifically inhibited by unlabeled PDGF, and saturable. Scatchard plot analysis showed a single class of sites with 3000 +/- 450 molecules bound/cell and an apparent KD of 1.2 +/- 0.2 10(-8) M. The effects of PDGF on collagen-induced phosphoinositide breakdown and protein phosphorylation were also investigated. At 50 ng/ml PDGF, a concentration which completely inhibited collagen-induced aggregation, the breakdown of (/sup 32/P)phosphatidylinositol 4,5-biphosphate (PIP2) and (/sup 32/P)phosphatidylinositol 4-phosphate (PIP) was observed, but the subsequent replenishment of (/sup 32/P)PIP2 was inhibited. The same PDGF concentration totally inhibited collagen-induced phosphatidic acid formation. PDGF also completely prevented phosphorylation of P43 and P20, as a result of protein kinase C activation consecutive to phosphoinositide metabolism. These results suggest that a specific PDGF receptor can be induced by collagen, and PDGF can effect the early events of collagen-induced platelet activation by inhibiting PIP2 resynthesis and P43 and P20 phosphorylation. It is concluded that PDGF might be involved in a negative feed-back control of platelet activation.

  20. In vitro activation of a soluble cholesterol esterase from bovine adrenals by a cAMP-dependent protein kinase.

    PubMed

    Wallat, S; Kunau, W H

    1976-07-01

    Properties and partial purification of the bovine adrenal cholesterol esterase from the 100000 X g supernatant fraction were investigated. Variations of the enzyme activity with time-dependent (enzymatic) and time-dependent (non enzymatic) effects have been demonstrated. Mg2 has been proved to inhibit the enzyme activity by a non-enzymatic effect in 50mM Tris/HCl buffer, pH 7.4. A time-dependent inactivation of the cholesterol esterase has been observed in the same buffer. The enzyme could be protected from this enzymatic inactivation by its substrate, cholesterol oleate. cAMP, ATP and Mg2 cuase a time-dependent stimulation of the enzyme in 50mM Tris/HCl buffer, pH 7.4. This result suggests that corticotropin activates the soluble cholesterol esterase from bovine adrenals via cAMP-dependent protein kinase. This view is strengthened by the incorporation of 32P radioactivity from [gamma-32P] ATP into the protein fraction of the 100,000 X g supernatant. The protein-bound 32P radioactivity could be co-purified with the enzyme activity during the partial purification of the soluble cholesterol esterase.