Sample records for autoradiography

  1. Autoradiography and the Cell Cycle. (United States)

    Jones, C. Weldon


    Outlines the stages of a cell biology "pulse-chase" experiment in which the students apply autoradiography techniques to learn about the concept of the cell cycle. Includes (1) seed germination and plant growth; (2) radioactive labeling and fixation of root tips; (3) feulgen staining of root tips; (4) preparation of autoradiograms; and…

  2. Digital autoradiography using silicon strip detectors

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    Overdick, M.


    Spatially resolving radiation detection systems operating in real time can be used to acquire autoradiographic images. An overview over alternatives to traditional autoradiography is given and the special features of these filmless methods are discussed. On this basis the design of a system for digital autoradiography using silicon strip detectors is presented. Special emphasis is put on the physical background of the detection process in the semiconductor and on the self-triggering read-out technique. The practical performance of the system is analyzed with respect to energy and spatial resolution. This analysis is complemented by case studies from cell biology (especially electrophoresis), botany and mineralogy. Also the results from a time-resolved autoradiographic experiment are presented. (orig.) 80 refs.


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    Gibbs, K; Carol Kestin, C


    This report fulfills the FY 2006 Enhanced Surveillance Campaign Level 3 milestones for Task TSR 11.1 as defined in the execution plan [1, 2]. The purpose of this task is to reduce the cycle time necessary to complete analytical evaluations required for surveillance of reservoirs. The development of the digital autoradiography system supports this task. The digital autoradiography system is currently operational and ready for implementation in reservoir surveillance performed in the Materials Test Facility (MTF) at Savannah River Site (SRS). SRS requests design agency (Los Alamos National Laboratory and Sandia National Laboratory) concurrence for the implementation of this system and on the establishment, in conjunction with the Savannah River National Laboratory (SRNL), of the implementation requirements for this system. Stainless steel tritium reservoirs and pinch welded tubes, which have been exposed to tritium for a prolonged period, are destructively evaluated at the end of their service lives for a variety of reasons. One requirement of this evaluation is to assess the tritium diffusion into the reservoir material. The current method used to determine the geometry and depth of tritium penetration is autoradiography. This technique employs a photographic emulsion and has been effective for a number of years. The primary disadvantage of this technique is the time required to obtain results. The success of the traditional technique is dependent on many variables, such as the proficiency of the operators in conducting sample preparation, the geometry of the sample and the shelf life of the photographic chemicals. If results are not satisfactory, several repetitions are often required and usually add weeks to the total analysis time for the sample. Due to the extensive time required for the liquid emulsion autoradiography method, a new, faster technique was desired. Personnel from the Savannah River National Laboratory (SRNL) have been working on a system based on

  4. Autoradiography of manganese: accumulation and retention in the pancreas

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    Lyden, A.; Lindquist, N.G. (National Board of Health and Welfare, Department of Drugs, Division of Pharmacology and Toxicology, Uppsala, Sweden); Larsson, B.S. (Department of Toxicology, University of Uppsala, Biomedical Center, Uppsala, Sweden)


    By means of whole-body autoradiography, the general distribution of /sup 54/MnCl/sub 2/ was studied in mice and a Marmoset monkey. High accumulation and retention were observed in the pancreas in both species. Gamma counting experiments in mice after a single intravenous injection of /sup 54/MnCl/sub 2/ showed that the level in the pancreas exceeded that of the liver at all survival times (20 min. - 30 days). Also in the monkey, the concentration in the pancreas exceeded that of the liver, and the pancreas had the highest tissue/liver ratio of the organs measured at 24 hours after injection. The high uptake and long retention in the pancreas suggest that manganese is of importance for the pancreatic function but also that the pancreas may be a target organ for manganese toxicity. Positron tomography, using /sup 11/C-labelled amino acids, has been found to be a promising diagnostic technique for the study of pancreatic disease. Positron emitting manganese isotopes may be worth further studies as possible agents for pancreatic imaging.

  5. Neutron induced autoradiography of some minerals from the Allchar mine

    CERN Document Server

    Lazaru, A; Skvarc, J; Kristof, E S; Stafilov, T


    The mineral lorandite from the Allchar mine (Kavadarci, Macedonia) will be used to estimate the average solar neutrino flux. Here, the amount of sup 2 sup 0 sup 5 Pb isotope induced by the sup 2 sup 0 sup 5 Tl(nu sub e , e sup -) sup 2 sup 0 sup 5 Pb reaction is measured and converted to neutrino flux. To determine the few sup 2 sup 0 sup 5 Pb atoms that are produced by solar neutrinos in the Tl ore it is necessary to know all the interfering reactions and/or impurities producing sup 2 sup 0 sup 5 Pb. The concentration and/or spatial distribution of some impurities such as U in lorandite should be known as accurately as possible. In the present work uranium and boron concentrations in some minerals from the Allchar mine (lorandite, realgar, stibnite, orpiment and dolomite) were measured by neutron induced autoradiography. The tracks of sup 1 sup 0 B(n, alpha) and sup 2 sup 3 sup 5 U(n, f) reaction products were recorded by CR-39 and phosphate glass (PSK-50) etched track detectors, respectively. Results showed...

  6. Detection of androgen receptor in human prostatic adenoma by autoradiography

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    Demura, Takayoshi; Sakashita, Shigeo; Takamura, Takao; Kuroda, Kazuhide (Asahikawa Medical Coll., Hokkaido (Japan))


    We developed a new amplified method to detect the localization of androgen receptors within the human prostatic tissue specimens. The tissue sections were treated with 50 of 100 nM tritiated dihydrotestosterone (/sup 3/H-DHT). The binding of /sup 3/H-DHT to receptors was demonstrated as silver grains on the stained tissue sections. The binding of /sup 3/H-DHT to the prostatic tissue was inhibited by additional non-radioactive DHT remarkably and by testosterone partially, but not affected by additional progesterone and 17..beta..-estradiol. No binding of /sup 3/H-DHT to the bladder tissue was found. These results showed that the binding of /sup 3/H-DHT to the prostatic tissue was a specific reaction of /sup 3/H-DHT and androgen receptor. Androgen receptors were seen in the nuclei and the cytoplasmas of glandular epithelial cells of prostate. However, stromal cells contained less abundant androgen receptors. The method reported here has several advantages in detecting the androgen receptor of the prostatic tissue in comparison with the radioreceptor assay and other histochemical methods. 1) The needle biopsied specimens are big enough to examine. 2) Morphological observations are also possible on the same specimen because the specimens are stained with hematoxylin simultaneously. Therefore, we can know the relative ratio of androgen receptor positive cells and negative cells. 3) Binding of /sup 3/H-DHT to the receptor with this method may be more specific than other histochemical methods, since binding of /sup 3/H-DHT to the receptor was inhibited by 200-fold excess of non-radioactive DHT. 4) Treatment of scintillator, fluorographic technique shortens the exposure periods. The exposure periods are approximately six to twelve times shorter than that of the conventional autoradiography.

  7. Thymidine plaque autoradiography of thymidine kinase-positive and thymidine kinase-negative herpesviruses

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    Tenser, R.B.; Jones, J.C.; Ressel, S.J.; Fralish, F.A.


    Plaques formed by herpes simplex virus (HSV), pseudorabies virus, and varicella-zoster virus were studied by plaque autoradiography after (/sup 14/C)thymidine labeling. Standard thymidine kinase-positive (TK+) viruses and TK- mutants of HSV types 1 and 2 and pseudorabies virus were studied, including cell cultured viruses and viruses isolated from animals. Autoradiography was performed with X-ray film with an exposure time of 5 days. After development of films, TK+ plaques showed dark rims due to isotope incorporation, whereas TK- plaques were minimally labeled. Plaque autoradiography of stock TK- viruses showed reversion frequencies to the TK+ phenotype of less than 10(-3). Autoradiography indicated that TK- virus retained the TK- phenotype after replication in vivo. In addition, it was shown that TK- HSV could be isolated from mouse trigeminal ganglion tissue after corneal inoculation of TK- HSV together with TK+ HSV. The plaque autoradiographic procedure was very useful to evaluate proportions of TK+ and TK- virus present in TK+-TK- virus mixtures.

  8. Assignment of two ultrastructures formed by a mixture of hexonamides using autoradiography and electron microscopy

    NARCIS (Netherlands)

    Boettcher, Christoph; Boekema, Egbert J.; Fuhrhop, Juergen-H.


    The combined application of autoradiography and electron microscopy allowed the assignment of molecular components to individual micellar fibres in a mixed gel. Resolution was of the order of 0·1 µm. As a result, it was shown that bimolecular sheets of N-dodecyl-L-mannonamide (= L-Man-12) completely

  9. A Significant Contribution of INAA in Autoradiography for Elemental Profile Construction (United States)

    Khaweerat, Sasiphan; Ratanatongchai, Wichian; Channuie, Jatechan

    Autoradiography has been carried out occasionally at the out-of-pool neutron radiography (NR) facility of the Thai Research Reactor-1/M1 for more than a decade. Because of low and uncertain neutron flux at exposure position, the technique is not well recognized. The first attempt to conduct neutron activation autoradiography (NAAR) using in-pool irradiation facility was initiated in Thailand in early 2014. The technique combines fundamental concepts of instrumental neutron activation analysis (INAA) and autoradiography to fulfil their limitations. The elemental composition obtained from INAA, however, is for the whole sample and it is not possible to map specific location of particular elements. Autoradiography, on the other hand, provides visual images without elemental information. Here, a near triangle of 1.5 cm x 3.0 cm metal Buddha image was irradiated using the in-pool dry irradiation facility. Irradiation times were 10 seconds and 15 minutes and decay times were varied. In parallel to elemental analysis, the irradiated sample was placed in good contact with an imaging plate to record self- emitting radiation. At a condition of irradiation, autoradiographic image and elemental appearances in INAA spectrum were compared. The structural profile in complementary with elemental information will help address questions regarding provenance, manufacturing technology, traditional beliefs which can be further applied to conservation methodology and authenticity approval.

  10. Neutron autoradiography: working-out method and application in investigations of test paintings

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    Kalicki, A.; Panczyk, E.; Rowinska, L.; Sartowska, B. E-mail:; Walis, L.; Pytel, K.; Pytel, B.; Koziel, A.; Dabkowski, L.; Wierzchnicka, M.; Strzalkowski, L.; Ostrowski, T


    Neutron-induced autoradiography was carried out at MARIA research reactor in Poland. The paintings were exposed to the thermal neutrons. As a result, the radionuclides emitting beta particles and gamma rays were created from some of the elements existing in the painting. Beta particles were detected during successive exposure to a series of X-ray medical-sensitive films. The obtained images--blackening of the films depends mainly on the nuclear characteristic of recorded radionuclides and exposure parameters. The main purpose of this work was to work out a method, build a special stand and test sample paintings using neutron autoradiography. Samples of paintings were investigated and according to the obtained results, optimum test parameters have been selected: neutron irradiation conditions and autoradiographs exposure conditions.

  11. A study of Cs-137 spatial distribution in soil thin sections by digital autoradiography (United States)

    Korobova, Elena; Dogadkin, Nikolay; Shiryaev, Andrey; Kolotov, Vladimir; Turkov, Victor


    Recent studies have proved autoradiography to have high potential in detection of radiation in particles including geological objects [1-3]. We applied digital autoradiography based on usage of image plates to study Cs-137 microdistribution in thin sections of the podzolic sandy soil typical for the Chernobyl remote impact zone 25 years after the accident. The zone is noted for contamination of the so-called condensation type where the contribution of the "hot" fuel particles has been comparatively low. The initial 137Cs contamination level of the study plot approximated 40 Ci/km2. According to the soil core data twenty five years after the accident the main portion of cesium radioisotopes is still concentrated in the 10-20 cm thick surface layer. Thin sections have been prepared from the top 0-10 cm soil layer of the soil profile located on the shoulder of the relatively steep northern slope of the forested hill formed on the Iput river terrace ca 20 km to the east of the town of Novozybkov, Bryansk region. Undisturbed soil sample was impregnated with epoxy resin, then dissected in vertical triplicates and polished to obtain open surface. Autoradiography of the thin sections has clearly shown different patterns of Cs-137 distribution related to its concentration in organic material and on the surface of soil particles. High sensitivity and resolution of the applied technique enables to reveal concentration and dispersion zones on microscale level. Soil micro-morphology has shown to be helpful in deciphering soil components and properties responsible for Cs-137 retention in the soil top layer. References 1. Mihoko Hareyama, Noriyoshi Tsuchiya, Masahiro Takebe and Tadashi Chida. Two-dimensional measurement of natural radioactivity of granitic rocks by photostimulated luminescence technique Geochemical Journal, 2000, 34, 1- 9. 2. Zeissler C. J., R. M. Lindstrom, J. P. McKinley. Radioactive particle analysis by digital autoradiography. Journal of Radioanalytical and

  12. Characterization of a double-sided silicon strip detector autoradiography system

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    Örbom, Anders, E-mail:; Ahlstedt, Jonas; Östlund, Karl; Strand, Sven-Erik [Department of Medical Radiation Physics, Lund University, Lund SE-22185 (Sweden); Serén, Tom; Auterinen, Iiro; Kotiluoto, Petri [VTT Technical Research Centre of Finland, Espoo FI-02044 (Finland); Hauge, Håvard [Biomolex AS, Oslo NO-0319 (Norway); Olafsen, Tove; Wu, Anna M.; Dahlbom, Magnus [Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, California 90095 (United States)


    Purpose: The most commonly used technology currently used for autoradiography is storage phosphor screens, which has many benefits such as a large field of view but lacks particle-counting detection of the time and energy of each detected radionuclide decay. A number of alternative designs, using either solid state or scintillator detectors, have been developed to address these issues. The aim of this study is to characterize the imaging performance of one such instrument, a double-sided silicon strip detector (DSSD) system for digital autoradiography. A novel aspect of this work is that the instrument, in contrast to previous prototype systems using the same detector type, provides the ability for user accessible imaging with higher throughput. Studies were performed to compare its spatial resolution to that of storage phosphor screens and test the implementation of multiradionuclide ex vivo imaging in a mouse preclinical animal study. Methods: Detector background counts were determined by measuring a nonradioactive sample slide for 52 h. Energy spectra and detection efficiency were measured for seven commonly used radionuclides under representative conditions for tissue imaging. System dead time was measured by imaging {sup 18}F samples of at least 5 kBq and studying the changes in count rate over time. A line source of {sup 58}Co was manufactured by irradiating a 10 μm nickel wire with fast neutrons in a research reactor. Samples of this wire were imaged in both the DSSD and storage phosphor screen systems and the full width at half maximum (FWHM) measured for the line profiles. Multiradionuclide imaging was employed in a two animal study to examine the intratumoral distribution of a {sup 125}I-labeled monoclonal antibody and a {sup 131}I-labeled engineered fragment (diabody) injected in the same mouse, both targeting carcinoembryonic antigen. Results: Detector background was 1.81 × 10{sup −6} counts per second per 50 × 50 μm pixel. Energy spectra and

  13. Evaluation of thermal neutron irradiation field using a cyclotron-based neutron source for alpha autoradiography. (United States)

    Tanaka, H; Sakurai, Y; Suzuki, M; Masunaga, S; Mitsumoto, T; Kinashi, Y; Kondo, N; Narabayashi, M; Nakagawa, Y; Watanabe, T; Fujimoto, N; Maruhashi, A; Ono, K


    It is important to measure the microdistribution of (10)B in a cell to predict the cell-killing effect of new boron compounds in the field of boron neutron capture therapy. Alpha autoradiography has generally been used to detect the microdistribution of (10)B in a cell. Although it has been performed using a reactor-based neutron source, the realization of an accelerator-based thermal neutron irradiation field is anticipated because of its easy installation at any location and stable operation. Therefore, we propose a method using a cyclotron-based epithermal neutron source in combination with a water phantom to produce a thermal neutron irradiation field for alpha autoradiography. This system can supply a uniform thermal neutron field with an intensity of 1.7×10(9) (cm(-2)s(-1)) and an area of 40mm in diameter. In this paper, we give an overview of our proposed system and describe a demonstration test using a mouse liver sample injected with 500mg/kg of boronophenyl-alanine.

  14. Cholinergic Depletion in Alzheimer’s Disease Shown by [18F]FEOBV Autoradiography

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    Maxime J. Parent


    Full Text Available Rationale. Alzheimer’s Disease (AD is a neurodegenerative condition characterized in part by deficits in cholinergic basalocortical and septohippocampal pathways. [18F]Fluoroethoxybenzovesamicol ([18F]FEOBV, a Positron Emission Tomography ligand for the vesicular acetylcholine transporter (VAChT, is a potential molecular agent to investigate brain diseases associated with presynaptic cholinergic losses. Purpose. To demonstrate this potential, we carried out an [18F]FEOBV autoradiography study to compare postmortem brain tissues from AD patients to those of age-matched controls. Methods. [18F]FEOBV autoradiography binding, defined as the ratio between regional grey and white matter, was estimated in the hippocampus (13 controls, 8 AD and prefrontal cortex (13 controls, 11 AD. Results. [18F]FEOBV binding was decreased by 33% in prefrontal cortex, 25% in CA3, and 20% in CA1. No changes were detected in the dentate gyrus of the hippocampus, possibly because of sprouting or upregulation toward the resilient glutamatergic neurons of the dentate gyrus. Conclusion. This is the first demonstration of [18F]FEOBV focal binding changes in cholinergic projections to the cortex and hippocampus in AD. Such cholinergic synaptic (and more specifically VAChT alterations, in line with the selective basalocortical and septohippocampal cholinergic losses documented in AD, indicate that [18F]FEOBV is indeed a promising ligand to explore cholinergic abnormalities in vivo.

  15. {sup 10}B concentration evaluation in autoradiography images by optical density measurements

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    Portu, A., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)] [Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Rivadavia 1917, AC: C1033AAJ, Ciudad Autonoma de Buenos Aires (Argentina); Saint Martin, G., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina); Brandizzi, D., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)] [Laboratorio de Microespectrofotometria (LANAIS-MEF), CONICET-CNEA (Argentina); Bernaola, O.A., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)


    The quantification and analysis of the tracks forming the autoradiography image of a tissue section is essential for the measurement of particle emitter concentration and distribution (e.g. {sup 10}B) in the sample. Conventional counting techniques cannot be used when track density is high because of track overlapping. A densitometry supported by image analysis method suitable for these cases has been developed. Optical density measurements obtained for boron solutions of known concentrations showed a linear behavior in the range of concentrations under consideration. - Highlights: Black-Right-Pointing-Pointer A nuclear tracks quantification method was developed applying optical densitometry supported by image analysis. Black-Right-Pointing-Pointer It will provide information about {sup 10}B distribution in samples whose autoradiographies present high density of tracks. Black-Right-Pointing-Pointer A calibration curve for optical density versus boron concentration was constructed and applied to the analysis of boron-doped silicon wafers. Black-Right-Pointing-Pointer Results were compared with those from an individual counting technique, obtaining a good agreement.

  16. Reference systems for the determination of {sup 10}B through autoradiography images: Application to a melanoma experimental model

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    Portu, A., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)] [Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Rivadavia 1917, AC: C1033AAJ, Ciudad Autonoma de Buenos Aires (Argentina); Carpano, M., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina); Dagrosa, A., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)] [Consejo Nacional de Investigaciones Cientificas y Tecnicas (CONICET), Rivadavia 1917, AC: C1033AAJ, Ciudad Autonoma de Buenos Aires (Argentina); Nievas, S., E-mail: [Centro Atomico Constituyentes (CAC), Comision Nacional de Energia Atomica (CNEA), Av. Gral. Paz 1499, AC: B1650KNA, San Martin, Buenos Aires (Argentina)


    The amount of {sup 10}B in tissue samples may be determined by measuring the track density in the autoradiography image produced on a nuclear track detector. Different systems were evaluated as reference standards to be used for a quantitative evaluation of boron concentration. The obtained calibration curves were applied to evaluate the concentration of {sup 10}B in melanoma tumour of NIH nude mice after a biodistribution study. The histological features observed in the tissue sections were accurately reproduced by the autoradiography images.

  17. Opioid binding sites in the guinea pig and rat kidney: Radioligand homogenate binding and autoradiography

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    Dissanayake, V.U.; Hughes, J.; Hunter, J.C. (Parke-Davis Research Unit, Addenbrookes Hospital Site, Cambridge (England))


    The specific binding of the selective {mu}-, {delta}-, and {kappa}-opioid ligands (3H)(D-Ala2,MePhe4,Gly-ol5)enkephalin ((3H) DAGOL), (3H)(D-Pen2,D-Pen5)enkephalin ((3H)DPDPE), and (3H)U69593, respectively, to crude membranes of the guinea pig and rat whole kidney, kidney cortex, and kidney medulla was investigated. In addition, the distribution of specific 3H-opioid binding sites in the guinea pig and rat kidney was visualized by autoradiography. Homogenate binding and autoradiography demonstrated the absence of {mu}- and {kappa}-opioid binding sites in the guinea pig kidney. No opioid binding sites were demonstrable in the rat kidney. In the guinea pig whole kidney, cortex, and medulla, saturation studies demonstrated that (3H)DPDPE bound with high affinity (KD = 2.6-3.5 nM) to an apparently homogeneous population of binding sites (Bmax = 8.4-30 fmol/mg of protein). Competition studies using several opioid compounds confirmed the nature of the {delta}-opioid binding site. Autoradiography experiments demonstrated that specific (3H)DPDPE binding sites were distributed radially in regions of the inner and outer medulla and at the corticomedullary junction of the guinea pig kidney. Computer-assisted image analysis of saturation data yielded KD values (4.5-5.0 nM) that were in good agreement with those obtained from the homogenate binding studies. Further investigation of the {delta}-opioid binding site in medulla homogenates, using agonist ((3H)DPDPE) and antagonist ((3H)diprenorphine) binding in the presence of Na+, Mg2+, and nucleotides, suggested that the {delta}-opioid site is linked to a second messenger system via a GTP-binding protein. Further studies are required to establish the precise localization of the {delta} binding site in the guinea pig kidney and to determine the nature of the second messenger linked to the GTP-binding protein in the medulla.

  18. Quantitative autoradiography of (/sup 3/H)corticosterone receptors in rat brain

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    Sapolsky, R.M.; McEwen, B.S. (Rockefeller Univ., New York (USA)); Rainbow, T.C. (Pennsylvania Univ., Philadelphia (USA). School of Medicine)


    The authors have quantified corticosterone receptors in rat brain by optical density measurements of tritium-film autoradiograms. Rats were injected i.v. with 500 (/sup 3/H)corticosterone to label brain receptors. Frozen sections of brain were cut with a cryostat and exposed for 2 months against tritium-sensitive sheet film (LKB Ultrofilm). Tritium standards were used to convert optical density readings into molar concentrations of receptor. High levels of corticosterone receptors were present throughout the pyramidal and granule cell layers of the hippocampus. Moderate levels of receptors were found in the neuropil of the hippocampus, the lateral septum, the cortical nucleus of the amygdala and the entorhinal cortex. All other brain regions had low levels of receptors. These results extend previous non-quantitative autoradiographic studies of corticosterone receptors and provide a general procedure for the quantitative autoradiography of steroid hormone receptors in brain tissue.

  19. Autoradiography of geological fluorite samples for determination of uranium and thorium distribution using nuclear track methodology

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    Pi, T.; Sole, J. [Instituto de Geologia, UNAM, Cd. Universitaria, Coyoacan, 04510 Mexico DF (Mexico); Golzarri, J.I; Rickards, J.; Espinosa, G. [IFUNAM, AP 20-364, 01000 Mexico DF (Mexico)]. e-mail:


    In this paper we present the uranium and thorium distribution analysis of several samples of the 'La Azul' an epithermal fluorspar deposit in southern Mexico, using nuclear track methodology (NTM), in the alpha-autoradiography mode, by placing the mineral sample in contact with a polycarbonate detector. This constitutes a non-destructive analysis, with sufficient sensitivity to provide valuable information about textural and para genetic characteristics of the geological samples. The selected nuclear track detector was CR-39 (Landauer). The region of interest of the geological samples was polished and put in contact with the detector material surface for 45 days in a vacuum chamber (10-3 torr). After this period of time, the detectors were chemically etched, revealing the auto radiograph of the radioactive material. The results show a clear distribution of bands of uranium and thorium in the fluorite samples. This is valuable information for the genetic or geochronological studies of the ore deposits. (Author)

  20. Alpha adrenergic receptors in dog coronary arteries as detected with autoradiography

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    Muntz, K.; Calianos, T.; Buja, L.M.


    The authors used previously established methods to determine the presence of alpha adrenergic receptors in different sizes of dog coronary arteries using autoradiography of /sup 3/H-prazosin (PRAZ) and /sup 125/I-BE 2254 (HEAT) to label alpha/sub 1/ adrenergic receptors and /sup 3/H-rauwolscine (RAUW) to label alpha/sub 2/ adrenergic receptors. Frozen sections of the left main coronary artery (LMA), the left anterior descending artery (LAD) and myocardium were incubated in 3 concentrations of PRAZ (0.1, 0.5 and 1.0 nM) (n=5 dogs), 3 concentrations of RAUW (1, 3 and 5 nM) (n=5) and one concentration of HEAT (50 pM) (n=3). All incubations were done in the absence of (total binding) or presence of (nonspecific binding) 10/sup -5/ M phentolamine or yohimbine. The sections were processed for autoradiography and silver grains quantitated using an image analyzer. Analysis of variance determined that there was a significant difference between total and nonspecific binding in the LMA incubated with PRAZ (p < 0.016), but no significant difference between total and nonspecific binding in the LAD (p < 0.19) or in the arterioles (p < 0.68). In the experiments with HEAT, similar results were obtained. With RAUW, there was significant labeling of arterioles (p < 0.004), but not over the LAD (p < 0.11) or the LMA (p < 0.49). The results suggest that the number of coronary alpha/sub 1/ receptors decreases as vessel size decreases, while the number of alpha/sub 2/ receptors increases as vessel size decreases.

  1. Down-regulation of rat kidney calcitonin receptors by salmon calcitonin infusion evidence by autoradiography

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    Bouizar, Z.; Rostene, W.H.; Milhaud, G.


    In treating age-related osteoporosis and Paget disease of bone, it is of major importance to avoid an escape phenomenon that would reduce effectiveness of the treatment. The factors involved in the loss of therapeutic efficacy with administration of large pharmacological doses of the hormone require special consideration. Down-regulation of the hormone receptors could account for the escape phenomenon. Specific binding sites for salmon calcitonin (sCT) were characterized and localized by autoradiography on rat kidney sections incubated with /sup 125/I-labeled sCT. Autoradiograms demonstrated a heterogeneous distribution of /sup 125/I-labeled sCT binding sites in the kidney, with high densities in both the superficial layer of the cortex and the outer medulla. Infusion of different doses of unlabeled sCT by means of Alzet minipumps for 7 days produced rapid changes in plasma calcium, phosphate, and magnesium levels, which were no longer observed after 2 or 6 days of treatment. Besides, infusion of high doses of sCT induced down-regulation of renal sCT binding sites located mainly in the medulla, where calcitonin (CT) has been shown to exert it physiological effects on water and ion reabsorption. These data suggest that the resistance to high doses of sCT often observed during long-term treatment of patients may be the consequence of not only bone-cell desensitization but also down-regulation of CT-sensitive kidney receptor sites.

  2. Quantitative autoradiography using a radioimager based on a multiwire proportional chamber (United States)

    Petegnief, Yolande; Petiet, Anne; Peker, Muammer Can; Bonnin, François; Meulemans, Alain; LeGuludec, Dominique


    Determination of the biodistribution of radiopharmaceuticals is an important issue for the evaluation of their performance in diagnosis and therapy. In this study, we evaluated a digital radioimager (RI) based on a multiwire proportional chamber for quantitative autoradiography (AR). The RI allows direct detection of electronic emissions of gamma emitters. Its qualitative and quantitative performances were tested on Tc and In labelled sections and compared with conventional film AR. Linearity of count rate versus activity was verified over a range of activity. As compared with film AR, a substantial improvement of the detection limit was obtained even for acquisition periods up to 20 times less than film exposure times. We provided the basis for quantitative analysis with tissue equivalent paste standards: the Tc and In RI counting efficiencies were respectively 1.19% and 2.35%. We illustrated the respective values of RI and film AR in two rat studies: Tc-DMSA in kidney and dual-isotope Tc-MIBI and In-antimyosin in heart. Calculated activity concentrations on sections of rat organs confirmed good correlation to gamma counting (deviation less than 12%). We suggest RI as a convenient technique for fast localization of single or dual-isotope tracers and determination of activity distribution.

  3. Quantitative autoradiography using a radioimager based on a multiwire proportional chamber

    Energy Technology Data Exchange (ETDEWEB)

    Petegnief, Y.; Peker, M.C.; Meulemans, A.; Le Guludec, D. [Service de Medecine Nucleaire et de Biophysique, Hopital Bichat-Claude Bernard, 46 rue H Huchard, F-75877 Paris Cedex 18 (France); Petiet, A.; Bonnin, F. [Laboratoire de Biophysique, Universite Paris 7, BP 416, F-75780 Paris (France)


    Determination of the biodistribution of radiopharmaceuticals is an important issue for the evaluation of their performance in diagnosis and therapy. In this study, we evaluated a digital radioimager (RI) based on a multiwire proportional chamber for quantitative autoradiography (AR). The RI allows direct detection of electronic emissions of gamma emitters. Its qualitative and quantitative performances were tested on {sup 99m}Tc and {sup 111}In labelled sections and compared with conventional film AR. Linearity of count rate versus activity was verified over a 10{sup 4} range of activity. As compared with film AR, a substantial improvement of the detection limit was obtained even for acquisition periods up to 20 times less than film exposure times. We provided the basis for quantitative analysis with tissue equivalent paste standards: the {sup 99m}Tc and {sup 111}In RI counting efficiencies were respectively 1.19% and 2.35%. We illustrated the respective values of RI and film AR in two rat studies: {sup 99m}Tc-DMSA in kidney and dual-isotope {sup 99m}Tc-MIBI and {sup 111}In-antimyosin in heart. Calculated activity concentrations on sections of rat organs confirmed good correlation to gamma counting (deviation less than 12%). We suggest RI as a convenient technique for fast localization of single or dual-isotope tracers and determination of activity distribution. (author)

  4. Polonium-210 in mussels and its implications for environmental alpha-autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    McDonald, P.; Baxter, M.S.; Fowler, S.W.; Heyraud, M.


    Alpha-autoradiographic and radiochemical studies of the distributions of transuranic nuclides in the tissues and organs of mussels collected from the vicinity of the British Nuclear Fuels reprocessing plant at Sellafield, England, appeared to require assessment also of baseline alpha-activities of natural /sup 210/ Po levels. To ensure that the levels of /sup 210/Po in Cumbrian mussels were not artificially enhanced by local discharges, mussels from remote British and French coastal sites were also analysed. General similarities in /sup 210/Po concentrations found in mussel soft parts suggest that the /sup 210/Po levels in the Ravenglass mussels are natural and largely unsupported by /sup 210/Pb; however these levels are as much as four times greater than the present day /sup 239 +240/Pu concentrations in the same samples. These findings severely limit the usefulness of alpha-autoradiography studies for transuranic nuclides performed on such samples. Because of the recently lower concentrations of alpha-emitting transuranic nuclides (mainly /sup 238/Pu, /sup 239 +240/Pu and /sup 241/Am) in the Ravenglass environment, natural /sup 210/Po is now a major contributor to alpha-track distributions in auto-radiographic studies.

  5. Autoradiography reveals selective changes in serotonin binding in neocortex of patients with temporal lobe epilepsy. (United States)

    Rocha, Luisa; Lorigados-Pedre, Lourdes; Orozco-Suárez, Sandra; Morales-Chacón, Lilia; Alonso-Vanegas, Mario; García-Maeso, Iván; Villeda-Hernández, Juana; Osorio-Rico, Laura; Estupiñán, Bárbara; Quintana, Christian


    The main goal of the present study was to evaluate binding to serotonin in the neocortex surrounding the epileptic focus of patients with mesial temporal lobe epilepsy (MTLE). Binding to 5-HT, 5-HT(1A), 5-HT(4), 5-HT(7) receptors and serotonin transporter (5-HTT) in T1-T2 gyri of 15 patients with MTLE and their correlations with clinical data, neuronal count and volume were determined. Autopsy material acquired from subjects without epilepsy (n=6) was used as control. The neocortex from MTLE patients demonstrated decreased cell count in layers III-IV (21%). No significant changes were detected on the neuronal volume. Autoradiography experiments showed the following results: reduced 5-HT and 5-HT(1A) binding in layers I-II (24% and 92%, respectively); enhanced 5-HT(4) binding in layers V-VI (32%); no significant changes in 5-HT(7) binding; reduced 5-HTT binding in all layers (I-II, 90.3%; III-IV, 90.3%, V-VI, 86.9%). Significant correlations were found between binding to 5-HT(4) and 5-HT(7) receptors and age of seizure onset, duration of epilepsy and duration of antiepileptic treatment. The present results support an impaired serotoninergic transmission in the neocortex surrounding the epileptic focus of patients with MTLE, a situation that could be involved in the initiation and propagation of seizure activity.

  6. The impact of atrazine on lake periphyton communities, including carbon uptake dynamics using track autoradiography. (United States)

    Hamilton, P B; Jackson, G S; Kaushik, N K; Solomon, K R


    Chlorophyll a, freshweight biomass, ash-free dry weight, cell numbers, species richness, community carbon uptake and species-specific carbon uptake were used to monitor the impact of atrazine (2 chloro-4-ethylamino-6-isopropylamino-s-triazine) on an in situ, enclosed periphyton community. Atrazine concentrations ranging from 0.08 to 1.56 mg litre(-1) were used during the 2 years of study. In both 1982 and 1983, there was a shift from a chlorophyte- to a diatom-dominated community. In 1982 the cyanobacterium Cylindrospermum stagnale and the chlorophyte Tetraspora cylindrica developed isolated colonies in the 1.56 mg litre(-1) treatment, indicating resistance to atrazine at this concentration. After atrazine exposure, community productivity was reduced by 21% to 82% in the low to high exposures, respectively. After day 21 productivity returned to control levels. It was shown, using track autoradiography, that the productivities of the larger algae Mougeotia sp., Oedogonium sp., Tolypothrix limbata and Epithemia turgida were the most affected, with reductions of 74.3% to 93.1% that of the controls. All the biotic measures indicated reduced growth after herbicide exposure.

  7. Quantitative autoradiography of angiotensin II receptors in the brain and kidney

    Energy Technology Data Exchange (ETDEWEB)

    Gehlert, D.R.


    The renin-angiotensin system is an important component in the regulation of systemic blood pressure. Angiotensin II is the principal effector peptide of this system. Interaction of angiotensin II with specific receptors can produce in several organic systems. When administered into the brain this octa-peptide produces a variety of responses including a stimulation of drinking, increased systemic blood pressure and several neuroendocrine responses. Its effects on the kidney include alterations in arteriolar resistance, mesangial cell contraction and a feedback inhibition of the release of renin. Since this peptide produces profound effects on homeostatis by an interaction with specific receptors, the quantitative technique of in vitro autoradiography was applied to localize receptor populations for angiotensin II. Specific binding sites for a radiolabeled form of angiotensin II were localized in various brain and kidney regions. In the rat brain high densities of angiotensin II receptors were observed in the paraventricular and suprachiasmatic nuclei of the hypothalamus, supraoptic nucleus and the posterior lobe of the pituitary, brain areas in which angiotensin II modified neuroendocrine functions.

  8. Quantitative distribution of angiotensin II binding sites in rat brain by autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Saavedra, J.M.; Israel, A.; Plunkett, L.M.; Kurihara, M.; Shigematsu, K.; Correa, F.M.


    Angiotensin II binding sites were localized and quantified in individual brain nuclei from single rats by incubation of tissue sections with 1 nM /sup 125/I-(Sar1)-angiotensin II, (/sup 3/H)-Ultrofilm autoradiography, computerized microdensitometry and comparison with /sup 125/I-standards. High angiotensin II binding was present in the circumventricular organs (organon vasculosum laminae terminalis, organon subfornicalis and area postrema), in selected hypothalamic nuclei (nuclei suprachiasmatis, periventricularis and paraventricularis) and in the nucleus tractus olfactorii lateralis, the nucleus preopticus medianus, the dorsal motor nucleus of the vagus and the nucleus tractus solitarii. High affinity (KA from 0.3 to 1.5 X 10(9) M-1) angiotensin II binding sites were demonstrated in the organon subfornicalis, the nucleus tractus solitarii and the area postrema after incubation of consecutive sections from single rat brains with /sup 125/I-(Sar1)-angiotensin II in concentrations from 100 pM to 5 nM. These results demonstrate and characterize brain binding sites for angiotensin II of variable high affinity binding both inside and outside the blood-brain barrier.

  9. Evaluation of the S phase distribution of flow cytometric DNA histograms by autoradiography and computer algorithms. (United States)

    Sheck, L E; Muirhead, K A; Horan, P K


    Cell sorting and tritiated thymidine autoradiography were used to define the distribution of S phase cells in flow cytometric DNA histograms obtained from exponential mouse lymphoma cells (L5178Y). The numbers of labeled S phase cells, autoradiographically determined from cells sorted at 2-channel intervals in the G1/early S and late S/G2M regions of the histogram, were compared with the numbers of computed S phase cells in comparable 2-channel intervals as predicted by several computer algorithms used to extract cell cycle phase distributions from DNA histograms. Polynomial and multirectangle algorithms gave computed estimates of total %S in close agreement with the tritiated thymidine labeling index for the cell population, while multi-Gaussian algorithms underestimated %S. Interval autoradiographic and algorithm studies confirmed these results in that no significant differences were found between the autoradiographic S phase distribution and S phase distributions calculated by the polynomial and multirectangle models. However, S phase cells were significantly underestimated in G1/early S by a constrained multi-Gaussian model and in both G1/early S and late S/G2 by an unconstrained multi-Gaussian model. For the particular cell line (L5178Y), staining protocol (mithramycin following ethanol fixation) and instrumentation (Coulter TPS-2 cell sorter) used in this study, close agreement between computed %S and tritiated thymidine labeling index was found to be a reliable indicator of an algorithm's success in resolving S phase cells in the G1/S and S/G2 transition regions of the DNA histograms.

  10. /sup 99m/Tc-DPD uptake in juvenile hemarthrosis. Scintimetry and autoradiography of the knee in dogs

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, E.S.; Hjortdal, V.E.; Noer, I.; Christensen, S.B.; Holm, I.E.; Buenger, C.


    The pathogenesis of subchondral bone lesions and growth plate affection in hemophilic arthropathy was studied in puppies by means of repeated regional /sup 99m/Tc-diphosphonate scintimetry and contact autoradiography. Unilateral hemarthrosis of the knee was induced by biweekly intraarticular injections of autologous blood for 12 weeks. Hemarthrosis caused an early (2 to 4 weeks) decrease in uptake of /sup 99m/Tc-diphosphonate in the juxtaarticular growth plates (ratio 0.7) and a delayed (8 to 10 weeks) increase in epiphyseal uptake (ratio 1.5). In a recovery phase after hemarthrosis, growth plate uptake returned to normal, while the epiphyseal uptake remained elevated for 8 to 10 weeks. By contact autoradiography, the growth plate uptake was localized to the calcification layer at the metaphyseal aspect of the growth plates, while the epiphyseal uptake mainly was seen in the thin subchondral and subsynovial bone layer and around osteophytes. The changes in uptake of /sup 99m/Tc-diphosphonate following hemarthrosis for 3 months were reversible and could be ascribed to the presence of synovial inflammation.

  11. NOTE: First images of a digital autoradiography system based on a Medipix2 hybrid silicon pixel detector (United States)

    Mettivier, Giovanni; Montesi, Maria Cristina; Russo, Paolo


    We present the first images of beta autoradiography obtained with the high-resolution hybrid pixel detector consisting of the Medipix2 single photon counting read-out chip bump-bonded to a 300 µm thick silicon pixel detector. This room temperature system has 256 × 256 square pixels of 55 µm pitch (total sensitive area of 14 × 14 mm2), with a double threshold discriminator and a 13-bit counter in each pixel. It is read out via a dedicated electronic interface and control software, also developed in the framework of the European Medipix2 Collaboration. Digital beta autoradiograms of 14C microscale standard strips (containing separate bands of increasing specific activity in the range 0.0038-32.9 kBq g-1) indicate system linearity down to a total background noise of 1.8 × 10-3 counts mm-2 s-1. The minimum detectable activity is estimated to be 0.012 Bq for 36 000 s exposure and 0.023 Bq for 10 800 s exposure. The measured minimum detection threshold is less than 1600 electrons (equivalent to about 6 keV Si). This real-time system for beta autoradiography offers lower pixel pitch and higher sensitive area than the previous Medipix1-based system. It has a 14C sensitivity better than that of micro channel plate based systems, which, however, shows higher spatial resolution and sensitive area.

  12. Boron concentration measurements by alpha spectrometry and quantitative neutron autoradiography in cells and tissues treated with different boronated formulations and administration protocols. (United States)

    Bortolussi, Silva; Ciani, Laura; Postuma, Ian; Protti, Nicoletta; Luca Reversi; Bruschi, Piero; Ferrari, Cinzia; Cansolino, Laura; Panza, Luigi; Ristori, Sandra; Altieri, Saverio


    The possibility to measure boron concentration with high precision in tissues that will be irradiated represents a fundamental step for a safe and effective BNCT treatment. In Pavia, two techniques have been used for this purpose, a quantitative method based on charged particles spectrometry and a boron biodistribution imaging based on neutron autoradiography. A quantitative method to determine boron concentration by neutron autoradiography has been recently set-up and calibrated for the measurement of biological samples, both solid and liquid, in the frame of the feasibility study of BNCT. This technique was calibrated and the obtained results were cross checked with those of α spectrometry, in order to validate them. The comparisons were performed using tissues taken form animals treated with different boron administration protocols. Subsequently the quantitative neutron autoradiography was employed to measure osteosarcoma cell samples treated with BPA and with new boronated formulations.

  13. Distribution of /sup 14/C-ochratoxin A in the mouse monitored by whole-body autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Fuchs, R.; Appelgren, L.-E.; Hult, K.


    The tissue distribution of /sup 14/C-labeled ochratoxin A was studied in mouse using whole-body autoradiography. The distribution was followed for 18 days after one single intravenous injection of 5, corresponding to 160-230 ng toxin/g body weight. Very long persistence of /sup 14/C-ochratoxin A in the circulation was noticed and the toxin was detected in the blood even after 18 days when the experiment was finished. The radioactivity in the kidney was unequallly distributed with a slightly higher concentration in the inner cortical and medullary parts. This was seen from 24 hrs and on after injection. Very high concentrations of radioactivity were found in the bile of treated animals. The radioactivity extracted from several sections was chemically characterized with thin-layer chromatography and was found to represent /sup 14/C-ochratoxin A.

  14. Carbon-14-ochratoxin A distribution in the Japanese quail (Coturnix coturnix japonica) monitored by whole body autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Fuchs, R.; Appelgren, L.E.; Hagelberg, S.; Hult, K.


    Tissue distribution of the nephrotoxic mycotoxin ochratoxin A was characterized in laying Japanese quail by whole body autoradiography and scintillation counting using /sup 14/C-labelled toxin. Periodically for 8 days after one intravenous injection of 14 microCi/bird, corresponding to 70 ng/g body weight, birds were killed, frozen, and sagittal sections of the whole body were placed on X-ray film. In general, the ochratoxin disappeared from the avian body rapidly. Specific retention of radioactivity was seen as a ring-like distribution in yolks and growing follicles. After sectioning, organs and intestinal contents were removed from carcasses in a frozen condition, homogenized, extracted, chromatographed, and the radioactivity in fractions was measured by scintillation spectroscopy. High concentrations of ochratoxin A were found in gastric intestinal contents, probably originating from toxin excreted in the bile.

  15. Tolerance to salinity in Cucumis with neutron activation and autoradiography of sodium in seedlings of selected species of the genus

    Energy Technology Data Exchange (ETDEWEB)

    Lathrop, B.L.


    Part I. Nineteen species of Cucumis were compared for tolerance to sodium chloride in solution and sand culture. C. sativus L., and C. hardwickii died at 270 meq/I NaCl, while C. melo L. can tolerate up to 340 meq/I. C. myriocarpus, C. ficifolius, C. membranifolius, and C. meeusii are the most tolerant, surviving up to about 500 meq/I NaCl. The initial response of all species to gradually increasing levels of salinization is a bluish-green deepening of foliage color and reduction in growth, followed by intervenal chlorosis of older leaves and more pronounced chlorosis of new growth. Death occurred first in the mesic species in about 12-18 days and was characterized by general wilting and necrosis. In more tolerant species which were generally more xeric and woody, death occurred in about 18-28 days as the leaf blades became necrotic on wilted petioles. Part II. A cyclotron generated proton beam was used to induce a high density thermal neutron flux from a NaI target for conversion of /sup 23/Na/sup 24/ to Na in living and dried seedlings of Cucumis sativus (sensitive to salinity) and C. myriocarpus (tolerant). There was little difference in movement of salt in the living seedlings of the cultivated and wild species of Cucumis during a 10 hour period following neutron activation and before autoradiography. Some suggestion of transport of Na in main vascular bundles occurred during this period in C. myriocarpus. The creation of /sup 24/Na in air-dried seedlings of this taxon that had been salinized also permitted autoradiography of sodium distribution and provided results similar to in vivo studies.

  16. Autoradiography: Biomedical applications. January, 1975-August, 1981 (citations from the International Information Service for the Physics and Engineering Communities Data Base). Report for January 75-August 81

    Energy Technology Data Exchange (ETDEWEB)


    This bibliography contains citations concerning the methods, equipment, and biomedical applications of autoradiography. Electron microscopy (EM) is discussed at length, and sample preparation for EM is detailed. Numerous biological, biochemical and biophysical studies are discussed. (Contains 84 citations fully indexed and including a title list.)

  17. Statistical parametric maps of {sup 18}F-FDG PET and 3-D autoradiography in the rat brain: a cross-validation study

    Energy Technology Data Exchange (ETDEWEB)

    Prieto, Elena; Marti-Climent, Josep M. [Clinica Universidad de Navarra, Nuclear Medicine Department, Pamplona (Spain); Collantes, Maria; Molinet, Francisco [Center for Applied Medical Research (CIMA) and Clinica Universidad de Navarra, Small Animal Imaging Research Unit, Pamplona (Spain); Delgado, Mercedes; Garcia-Garcia, Luis; Pozo, Miguel A. [Universidad Complutense de Madrid, Brain Mapping Unit, Madrid (Spain); Juri, Carlos [Center for Applied Medical Research (CIMA), Movement Disorders Group, Neurosciences Division, Pamplona (Spain); Clinica Universidad de Navarra, Department of Neurology and Neurosurgery, Pamplona (Spain); Centro de Investigacion Biomedica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Pamplona (Spain); Pontificia Universidad Catolica de Chile, Department of Neurology, Santiago (Chile); Fernandez-Valle, Maria E. [Universidad Complutense de Madrid, MRI Research Center, Madrid (Spain); Gago, Belen [Center for Applied Medical Research (CIMA), Movement Disorders Group, Neurosciences Division, Pamplona (Spain); Centro de Investigacion Biomedica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Pamplona (Spain); Obeso, Jose A. [Center for Applied Medical Research (CIMA), Movement Disorders Group, Neurosciences Division, Pamplona (Spain); Clinica Universidad de Navarra, Department of Neurology and Neurosurgery, Pamplona (Spain); Centro de Investigacion Biomedica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Pamplona (Spain); Penuelas, Ivan [Clinica Universidad de Navarra, Nuclear Medicine Department, Pamplona (Spain); Center for Applied Medical Research (CIMA) and Clinica Universidad de Navarra, Small Animal Imaging Research Unit, Pamplona (Spain)


    Although specific positron emission tomography (PET) scanners have been developed for small animals, spatial resolution remains one of the most critical technical limitations, particularly in the evaluation of the rodent brain. The purpose of the present study was to examine the reliability of voxel-based statistical analysis (Statistical Parametric Mapping, SPM) applied to {sup 18}F-fluorodeoxyglucose (FDG) PET images of the rat brain, acquired on a small animal PET not specifically designed for rodents. The gold standard for the validation of the PET results was the autoradiography of the same animals acquired under the same physiological conditions, reconstructed as a 3-D volume and analysed using SPM. Eleven rats were studied under two different conditions: conscious or under inhalatory anaesthesia during {sup 18}F-FDG uptake. All animals were studied in vivo under both conditions in a dedicated small animal Philips MOSAIC PET scanner and magnetic resonance images were obtained for subsequent spatial processing. Then, rats were randomly assigned to a conscious or anaesthetized group for postmortem autoradiography, and slices from each animal were aligned and stacked to create a 3-D autoradiographic volume. Finally, differences in {sup 18}F-FDG uptake between conscious and anaesthetized states were assessed from PET and autoradiography data by SPM analysis and results were compared. SPM results of PET and 3-D autoradiography are in good agreement and led to the detection of consistent cortical differences between the conscious and anaesthetized groups, particularly in the bilateral somatosensory cortices. However, SPM analysis of 3-D autoradiography also highlighted differences in the thalamus that were not detected with PET. This study demonstrates that any difference detected with SPM analysis of MOSAIC PET images of rat brain is detected also by the gold standard autoradiographic technique, confirming that this methodology provides reliable results, although

  18. MO-G-17A-09: Quantitative Autoradiography of Biopsy Specimens Extracted Under PET/CT Guidance

    Energy Technology Data Exchange (ETDEWEB)

    Fanchon, L; Carlin, S; Schmidtlein, C; Humm, J; Yorke, E; Solomon, S; Deasy, J; Kirov, A [Memorial Sloan Kettering Cancer Center, New York, New York (United States); Burger, I [University Hospital of Zurich, Zurich, Switzerland, Zurich (Switzerland)


    Purpose: To develop a procedure for accurate determination of PET tracer concentration with high spatial accuracy in situ by performing Quantitative Autoradiography of Biopsy Specimens (QABS) extracted under PET/CT guidance. Methods: Autoradiography (ARG) standards were produced from a gel loaded with a known concentration of FDG biopsied with 18G and 20G biopsy needles. Specimens obtained with these needles are generally cylindrical: up to 18 mm in length and about 0.8 and 0.6 mm in diameter respectively. These standards, with similar shape and density as biopsy specimens were used to generate ARG calibration curves.Quantitative ARG was performed to measure the activity concentration in biopsy specimens extracted from ten patients. The biopsy sites were determined according to PET/CT's obtained in the operating room. Additional CT scans were acquired with the needles in place to confirm correct needle placements. The ARG images were aligned with the needle tip in the PET/CT images using the open source CERR software. The mean SUV calculated from the specimen activities (SUVarg) were compared to that from PET (SUVpet) at the needle locations. Results: Calibration curves show that the relation between ARG signal and activity concentration in those standards is linear for the investigated range (up to 150 kBq/ml). The correlation coefficient of SUVarg with SUVpet is 0.74. Discrepancies between SUVarg and SUVpet can be attributed to the small size of the biopsy specimens compared to PET resolution. Conclusion: The calibration procedure using surrogate biopsy specimens provided a method for quantifying the activity within the biopsy cores obtained under FDG-PET guidance. QABS allows mapping the activity concentration in such biopsy specimens with a resolution of about 1mm. QABS is a promising tool for verification of biopsy adequacy by comparing specimen activity to that expected from the PET image. A portion of this research was funded by a research grant from

  19. Recent technologic developments on high-resolution beta imaging systems for quantitative autoradiography and double labeling applications

    CERN Document Server

    Barthe, N; Chatti, K; Coulon, P; Maitrejean, S; 10.1016/j.nima.2004.03.014


    Two novel beta imaging systems, particularly interesting in the field of radiopharmacology and molecular biology research, were developed these last years. (1) a beta imager was derived from research conducted by Pr Charpak at CERN. This parallel plate avalanche chamber is a direct detection system of beta radioactivity, which is particularly adapted for qualitative and quantitative autoradiography. With this detector, autoradiographic techniques can be performed with emitters such as /sup 99m/Tc because this radionuclide emits many low-energy electrons and the detector has a very low sensitivity to low-range gamma -rays. Its sensitivity (smallest activity detected: 0.007 cpm/mm/sup 2/ for /sup 3/H and 0.01 for /sup 14/C), linearity (over a dynamic range of 10/sup 4/) and spatial resolution (50 mu m for /sup 3/H or /sup 99m/Tc to 150 mu m for /sup 32/P or /sup 18/F ( beta /sup +/)) gives a real interest to this system as a new imaging device. Its principle of detection is based on the analysis of light emitte...

  20. Reproducibility of regional cerebral blood flow measurement at rest with autoradiography method using {sup 123}I-IMP and SPECT

    Energy Technology Data Exchange (ETDEWEB)

    Saito, Kyoko; Murata, Hajime; Maruno, Hirotaka; Mori, Kazuaki; Tsuchida, Shoichi [Toranomon Hospital, Tokyo (Japan)


    Reproducibility of regional cerebral blood flow (rCBF) values obtained by the autoradiography (ARG) method with one SPECT scan and one point arterial blood sampling at 10 min after {sup 123}I-N-isopropyl-p-iodoamphetamine (IMP) infusion was evaluated. The variation in the cross calibration factor between SPECT counts and well-type scintillation counts remained 2.2% for 13 months. The error in rCBF by sampling time difference was less than 3% when sampling time was within 10 min{+-}30 sec after infusion. There was a favorable correlation between the rCBF values in two separate resting scans measured by the ARG method in 10 patients (r=0.907). And the mean ratio of rCBF values by the second study to those by the first study in the cerebellum and cerebral lobes in 10 patients was 0.989{+-}0.056 (mean{+-}SD). There was no significant difference in rCBF values between two separate studies. Reproducibility of rCBF with the ARG method was proved to be good. The ARG method was thought to be useful for the assessment of treatment effects and clinical courses. (author)

  1. Mapping of odor-related neuronal activity in the olfactory bulb by high-resolution 2-deoxyglucose autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Lancet, D.; Greer, C.A.; Kauer, J.S.; Shepherd, G.M.


    The spatial distribution of odor-induced neuronal activity in the olfactory bulb, the first relay station of the olfactory pathway, is believed to reflect important aspects of chemosensory coding. We report here the application of high-resolution 2-deoxyglucose autoradiography to the mapping of spatial patterns of metabolic activity at the level of single neurons in the olfactory bulb. It was found that glomeruli, which are synaptic complexes containing the first synaptic relay, tend to be uniformly active or inactive during odor exposure. Differential 2-deoxyglucose uptake was also observed in the somata of projection neurons (mitral cells) and interneurons (periglomerular and granule cells). This confirms and extends our previous studies in which odor-specific laminar and focal uptake patterns were revealed by the conventional x-ray film 2-deoxyglucose method due to Sokoloff and colleagues (Sokoloff, L., Reivich, M., Kennedy, C., DesRosiers, M. H., Patlak, C. S., Pettigrew, K. D., Sakurada, O. and Shinohara, M. (1977) J. Neurochem. 28, 897-916). Based on results obtained by the two methods, it is suggested that the glomerulus as a whole serves as a functional unit of activity. The high-resolution results are interpreted in terms of the well-characterized synaptic organization of the olfactory bulb and also serve to illustrate the capability of the 2-deoxyglucose autoradiographic technique to map metabolic activity in single neurons of the vertebrate central nervous system.

  2. 64Cu-ATSM and 18FDG PET uptake and 64Cu-ATSM autoradiography in spontaneous canine tumors: comparison with pimonidazole hypoxia immunohistochemistry

    Directory of Open Access Journals (Sweden)

    Hansen Anders E


    Full Text Available Abstract Background The aim of this study was to compare 64Cu-diacetyl-bis(N4-methylsemicarbazone (64Cu-ATSM and 18FDG PET uptake characteristics and 64Cu-ATSM autoradiography to pimonidazole immunohistochemistry in spontaneous canine sarcomas and carcinomas. Methods Biopsies were collected from individual tumors between approximately 3 and 25 hours after the intravenous injection of 64Cu-ATSM and pimonidazole. 64Cu-ATSM autoradiography and pimonidazole immunostaining was performed on sectioned biopsies. Acquired 64Cu-ATSM autoradiography and pimonidazole images were rescaled, aligned and their distribution patterns compared. 64Cu-ATSM and 18FDG PET/CT scans were performed in a concurrent study and uptake characteristics were obtained for tumors where available. Results Maximum pimonidazole pixel value and mean pimonidazole labeled fraction was found to be strongly correlated to 18FDG PET uptake levels, whereas more varying results were obtained for the comparison to 64Cu-ATSM. In the case of the latter, uptake at scans performed 3 h post injection (pi generally showed strong positive correlated to pimonidazole uptake. Comparison of distribution patterns of pimonidazole immunohistochemistry and 64Cu-ATSM autoradiography yielded varying results. Significant positive correlations were mainly found in sections displaying a heterogeneous distribution of tracers. Conclusions Tumors with high levels of pimonidazole staining generally displayed high uptake of 18FDG and 64Cu-ATSM (3 h pi.. Similar regional distribution of 64Cu-ATSM and pimonidazole was observed in most heterogeneous tumor regions. However, tumor and hypoxia level dependent differences may exist with regard to the hypoxia specificity of 64Cu-ATSM in canine tumors.

  3. Set-up and calibration of a method to measure 10B concentration in biological samples by neutron autoradiography (United States)

    Gadan, M. A.; Bortolussi, S.; Postuma, I.; Ballarini, F.; Bruschi, P.; Protti, N.; Santoro, D.; Stella, S.; Cansolino, L.; Clerici, A.; Ferrari, C.; Zonta, A.; Zonta, C.; Altieri, S.


    A selective uptake of boron in the tumor is the base of Boron Neutron Capture Therapy, which can destroy the tumor substantially sparing the normal tissue. In order to deliver a lethal dose to the tumor, keeping the dose absorbed by normal tissues below the tolerance level, it is mandatory to know the 10B concentration present in each kind of tissue at the moment of irradiation. This work presents the calibration procedure adopted for a boron concentration measurement method based on neutron autoradiography, where biological samples are deposited on sensitive films and irradiated in the thermal column of the TRIGA reactor (University of Pavia). The latent tracks produced in the film by the charged particles coming from the neutron capture in 10B are made visible by a proper etching, allowing the measurement of the track density. A calibration procedure with standard samples provides curves of track density as a function of boron concentration, to be used in the measurement of biological samples. In this paper, the bulk etch rate parameter and the calibration curves obtained for both liquid samples and biological tissues with known boron concentration are presented. A bulk etch rate value of (1.64 ± 0.02) μm/h and a linear dependence with etching time were found. The plots representing the track density versus the boron concentration in a range between 5 and 50 μg/g (ppm) are linear, with an angular coefficient of (1.614 ± 0.169)·10-3 tracks/(μm2 ppm) for liquids and (1.598 ± 0.097)·10-2 tracks/(μm2 ppm) for tissues.

  4. Binding of lurasidone, a novel antipsychotic, to rat 5-HT7 receptor: analysis by [3H]SB-269970 autoradiography. (United States)

    Horisawa, Tomoko; Ishiyama, Takeo; Ono, Michiko; Ishibashi, Tadashi; Taiji, Mutsuo


    Lurasidone is a novel antipsychotic agent with high affinity for dopamine D(2) and serotonin 5-HT(7), 5-HT(2A), and 5-HT(1A) receptors. We previously reported that in addition to its antipsychotic action, lurasidone shows beneficial effects on mood and cognition in rats, likely through 5-HT(7) receptor antagonistic actions. In this study, we evaluated binding of lurasidone to 5-HT(7) receptors in the rat brain by autoradiography using [(3)H]SB-269970, a specific radioligand for 5-HT(7) receptors. Brain slices were incubated with 4 nM [(3)H]SB-269970 at room temperature and exposed to imaging plates for 8 weeks before phosphorimager analysis. Using this method, we first investigated 5-HT(7) receptor distribution. We found that 5-HT(7) receptors are abundantly localized in brain limbic structures, including the lateral septum, thalamus, hypothalamus, hippocampus, and amygdala. On the other hand, its distribution was moderate in the cortex and low in the caudate putamen and cerebellum. Secondly, binding of lurasidone, a selective 5-HT(7) receptor antagonist SB-656104-A and an atypical antipsychotic olanzapine to this receptor was examined. Lurasidone, SB-656104-A (10–1000 nM), and olanzapine (100–10,000 nM) showed concentration-dependent inhibition of [(3)H]SB-269970 binding with IC(50) values of 90, 49, and 5200 nM, respectively. Similar inhibitory actions of these drugs were shown in in vitro [(3)H]SB-269970 binding to 5-HT(7) receptors expressed in Chinese hamster ovary cells. Since there was no marked species difference in rat and human 5-HT(7) receptor binding by lurasidone (K(i) = 1.55 and 2.10 nM, respectively), these findings suggest that binding to 5-HT(7) receptors might play some role in its beneficial pharmacological actions in schizophrenic patients.

  5. Mapping the distribution of serotonin transporter in the human brainstem with high-resolution PET: Validation using postmortem autoradiography data. (United States)

    Fazio, P; Schain, M; Varnäs, K; Halldin, C; Farde, L; Varrone, A


    The human brainstem is a complex structure with several small nuclei and neural pathways of interest in the pathophysiology of central nervous system (CNS) disorders. In common with other monoaminergic systems, serotoninergic neurons originate from a group of nuclei located in the brainstem. The present study was designed to validate a user-independent approach for a detailed in vivo quantification of serotonin transporter (5-HTT) availability in the human brainstem using a template-based approach that consisted of three steps. First, 3T-MR images and parametric binding potential (BPND) [(11)C]MADAM images of ten healthy subjects were used to generate a PET template of 5-HTT availability. In the second step, volumes of interest (VOIs) for different brainstem nuclei were obtained using a method in which VOIs are initially delineated on MRI images using anatomical landmarks and then are finally tailored on the distribution of 5-HTT binding using a thresholding approach applied to the 5-HTT template. In the final step, the VOIs were transformed and applied individually to BPND images of 16 healthy subjects (14M/2F, 20-64years). The in vivo distribution of BPND values obtained with the template-based method were in good agreement with an individual-based approach taken as gold standard. Results were also in agreement with 5-HTT quantification using in vitro binding data obtained with autoradiography (ARG) studies using [(3)H]MADAM. The proposed template-based method can be applied to PET data acquired in several CNS disorders in which serotonin neurons in the brainstem might be affected.

  6. Digital Autoradiography as a novel complementary technique for the investigation of radioactive contamination in nuclear facilities under dismantlement

    Energy Technology Data Exchange (ETDEWEB)

    Haudebourg, Raphael; Fichet, Pascal [Laboratory of Analyses and Operators Support, French Atomic Energy and Alternative Energies Commision (CEA), Saclay Center, 91191 Gif Sur Yvette Cedex, (France); Goutelard, Florence [NucLab, CEA Marcoule Center, (France)


    The detection (location and quantification) of nuclear facilities to be dismantled possible contamination with low-range particles emitters ({sup 3}H, other low-energy β emitters, a emitters) remains a tedious and expensive task. Indeed, usual remote counters show a too low sensitivity to these non-penetrating radiations, while conventional wipe tests are irrelevant for fixed radioactivity evaluation. The only method to accurately measure activity levels consists in sampling and running advanced laboratory analyses (spectroscopy, liquid scintillation counting, pyrolysis...). Such measurements generally induce sample preparation, waste production (destructive analyses, solvents), nuclear material transportation, long durations, and significant labor mobilization. Therefore, the search for the limitation of their number and cost easily conflicts with the necessity to perform a dense screening for sampling (to maximize the representativeness of the samples), in installations of thousands of square meters (floors, wells, ceilings), plus furniture, pipes, and other wastes. To overcome this contradiction, Digital Autoradiography (D. A.) was re-routed from bio molecular research to radiological mapping of nuclear installations under dismantling and to waste and sample analysis. After in-situ exposure to the possibly-contaminated areas to investigate, commercial reusable radiosensitive phosphor screens (of a few 100 cm{sup 2}) were scanned in the proper laboratory device and sharp quantitative images of the radioactivity could be obtained. The implementation of geostatistical tools in the data processing software enabled the exhaustive characterization of concrete floors at a rate of 2 weeks / 100 m{sup 2}, at lowest costs. Various samples such as drilled cores, or tank and wood pieces, were also successfully evaluated with this method, for decisive results. Thanks to the accurate location of potential contamination spots, this approach ensures relevant and representative

  7. Preclinical testing of F-18 and I-123 radioligands involving autoradiography and a phosphor-imaging device

    Energy Technology Data Exchange (ETDEWEB)

    Gatley, S.J.; Pyatt, B.; Gifford, A.N. [Brookhaven National Lab., Upton, NY (United States)] [and others


    During the development and validation of a receptor-binding radiopharmaceutical it is necessary to assess its brain regional distribution. Film contact autoradiography (AR) points such distributions to be evaluated on a fine anatomical scale, and avoids the inherent risk a microdissection experiments of missing interesting localization pattern. However, in vivo film AR may require a 1000-fold higher activity of a F-18 or I-123 radioligand than microdissection. In addition to health physics considerations, problems with receptor saturation can easily occur after injecting, say, 10 mCi into a 250g rat. A less commonly appreciated problem even with in vitro AR, using slide-mounted brain sections, is that it may be impossible to use sufficient F-18 to obtain acceptable images using film, because of receptor saturation. Phosphorimaging is superior in sensitivity to film by a factor of 10-50, and so relaxes constraints on injected activities and specific activities. Furthermore, it is also superior to film in terms of dynamic range and linearity of response. We have recently used phosphorimaging in two situations where film AR gave unsatisfactory results. (1) the novel cannabinoid CB1 receptor radioligand, I-123 AM281, was injected intravenously at 800 microcurie per rat. Animals were sacrificed at 2 h, frozen and whole body sections prepared. I-123 was clearly visualized in discrete brain areas (substantia nigra, globus pallidus, entopeduncular nucleus , hippocampus and cerebellum) consistent with the distribution of CB1 receptors previously demonstrated only in in vitro studies. (2) Rat brain sections were thaw mounted on gelatin/alum coated glass and incubated with the novel nicotinic acetylcholine receptor ligand F-18 2{prime}-fluoro-2{prime} nor-chloroepibatidine (1 nM).

  8. High-Resolution Autoradiography (United States)


    measurements see appendix ehrbide containced 6.68 percent carbon while the alpha iron 13). Theii Oak Ridge National Laboratory analysis gave (,t(e matrix...the cementite phase leaving only a (5) The sample was fixed for a minute, washed in water for very small amount of activity in the alpha iron phase. a

  9. Investigation of Figopitant and Its Metabolites in Rat Tissue by Combining Whole-Body Autoradiography with Liquid Extraction Surface Analysis Mass Spectrometry

    DEFF Research Database (Denmark)

    Schadt, S.; Kallbach, S.; Almeida, R.;


    This article describes the combination of whole-body autoradiography with liquid extraction surface analysis (LESA) and mass spectrometry (MS) to study the distribution of the tachykinin neurokinin-1 antagonist figopitant and its metabolites in tissue sections of rats after intravenous...... tissue extraction, sample cleanup, and high-performance liquid chromatography analysis. The parent drug and the N-dealkylated metabolite M474(1) (BIIF 1148) in varying ratios were the predominant compounds in all tissues investigated. In addition, several metabolites formed by oxygenation, dealkylation...

  10. Experimental studies on tissue distribution of /sup 14/C-labelled anitimicrobic drugs in otorhinolaryngological field. Special reference to apply macro-autoradiography to these studies

    Energy Technology Data Exchange (ETDEWEB)

    Murai, Kanetaka (Nagoya City Univ. (Japan). Faculty of Medicine)


    The distribution and concentration of antimicrobic drugs in the otorhinolaryngological field was studied together in the excised tissue by freezing macro-autoradiography in rabbits. Of the three Nalidixic acid antimicrobic analogues, i.e., pipemidic acid, AM-715 and rosoxacine, the first two accumulated remarkably in the soft palate, showing affinity for the cartilage. The concentrations of the three drugs in the paranasal sinus were higher on the medial portion of the maxilla than on the lateral portion, and the order of concentrations for ethmoidsinus were the lateral > anterior > posterior > medial portions. The present technique was thought to be a useful means for clarifying the distribution of drugs in vivo.

  11. A reproducible technique combining tritiated thymidine autoradiography with immunodetection of bromodeoxyuridine for double labelling studies of cell proliferation in paraffin sections of tissues. (United States)

    Hume, W J


    A method is described to combine tritiated thymidine autoradiography with immunoperoxidase detection of bromodeoxyuridine on the same paraffin sections. It overcomes the varied technical artefacts we encountered when first attempting to combine these techniques and results in preparations with extremely low peroxidase and autoradiographic backgrounds. In particular, we find it is important to avoid the use of detergents during immunostaining, otherwise grain counts are reduced and autoradiograph exposures need to be greatly increased, and to avoid excessive peroxidase staining which makes it difficult to visualize silver grains in the overlying emulsion. The advantages of a method to remove emulsion films using acid-alcohol, allowing the same sections to be dipped twice with a long and a short autoradiographic exposure, are presented. The routine combination of high quality tritiated thymidine autoradiography with clean immunoperoxidase staining of bromodeoxyuridine-positive nuclei provides a new and powerful cell kinetic, double-labelling method to augment existing techniques e.g. by labelling the same cells undergoing DNA synthesis in successive cell cycles.

  12. Recent technologic developments on high-resolution beta imaging systems for quantitative autoradiography and double labeling applications (United States)

    Barthe, N.; Chatti, K.; Coulon, P.; Maı̂trejean, S.; Basse-Cathalinat, B.


    Two novel beta imaging systems, particularly interesting in the field of radiopharmacology and molecular biology research, were developed these last years. (1) a beta imager was derived from research conducted by Pr Charpak at CERN. This parallel plate avalanche chamber is a direct detection system of β radioactivity, which is particularly adapted for qualitative and quantitative autoradiography. With this detector, autoradiographic techniques can be performed with emitters such as 99mTc because this radionuclide emits many low-energy electrons and the detector has a very low sensitivity to low-range γ-rays. Its sensitivity (smallest activity detected: 0.007 cpm/mm 2 for 3H and 0.01 for 14C), linearity (over a dynamic range of 10 4) and spatial resolution (50 μm for 3H or 99mTc to 150 μm for 32P or 18F (β +)) gives a real interest to this system as a new imaging device. Its principle of detection is based on the analysis of light emitted during the interaction with an intensified CCD camera. This property may suggest new potential applications, particularly in the field of β-rays selection according to their energy. This detector provides a new fast way to detect all β-emitting isotopes in biological samples up to 20 cm×25 cm (electrophoresis gels, hybridization membranes, tissue sections on glass slides, TLC plates and any other planar two-dimension samples). It is ideal for tritium detection, 500 times faster than classical film, thus maximizing the research productivity. (2) A micro imager is based on contact imaging through a solid scintillator sheet. Light emitted is amplified through an image intensifier tube and is analyzed with a CCD camera. The full field of view is smaller than the first one (24 mm×32 mm) but a better spatial resolution is obtained (typically 15 μm for 3H, 20 μm for 14C and 35S). The specifications of this detector are: efficiency 50-100% depending on isotope, linear response over a dynamic range of 10 4, smallest activity

  13. Quantitative autoradiography of ligands for dopamine receptors and transporters in brain of Göttingen minipig: comparison with results in vivo

    DEFF Research Database (Denmark)

    Minuzzi, Luciano; Alstrup, Aage Kristian Olsen; Bender, Dirk


    The pig has been used as animal model for positron emission tomography (PET) studies of dopamine (DA) receptors and pharmacological perturbations of DA neurotransmission. However, the binding properties of DA receptors and transporters in pig brain have not been characterized in vitro. Therefore......, the saturation binding parameters of [3H]SCH 23390 for DA D1 receptors and [3H]raclopride for DA D2/3 receptors were measured by quantitative autoradiography in cryostat sections from brain of groups of 8 week old and adult female Göttingen minipigs. The magnitudes of Bmax and Kd for these ligands were similar...... in young and old pigs, and were close to those reported for rat and human brain. Furthermore, gradients in the concentrations of D1 and D2/3 sites in striatum measured in vitro agreed with earlier findings in PET studies. However, the dopamine transporter (DAT) ligand [3H]GBR12935 did not bind in pig brain...

  14. Systemic injection of kainic acid: Gliosis in olfactory and limbic brain regions quantified with ( sup 3 H)PK 11195 binding autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Altar, C.A.; Baudry, M. (Genentech, Inc., South San Francisco, CA (USA))


    Neurodegenerative diseases may result from excessive stimulation of excitatory amino acid receptors by endogenous ligands. Because neuronal degeneration is associated with glial proliferation and hypertrophy, the degenerative changes throughout rat brain following the systemic administration of kainic acid (12 mg/kg) were mapped with quantitative autoradiography of (3H)PK 11195. This radioligand binds to a mitochondrial benzodiazepine binding site (MBBS) on microglia and astrocytes. Analysis of eight horizontal and four coronal brain levels revealed up to 16-fold increases in (3H)PK 11195 binding from 1 to 5 weeks but not 1 day after kainate injection. Increases in (3H)PK 11195 binding were predominantly in ventral limbic brain regions and olfactory projections to neocortical areas, with the olfactory cortex greater than subiculum/CA1 greater than anterior olfactory nucleus, medial thalamic nucleus, and piriform cortex greater than cingulate cortex and rostral hippocampus greater than dentate gyrus, septum, and amygdala greater than entorhinal cortex and temporal cortex. Little or no enhancement of (3H)PK 11195 binding was observed in numerous regions including the caudate-putamen, substantia nigra, nucleus accumbens, olfactory tubercle, cerebellum, thalamic nuclei, choroid plexus, medulla, parietal or occipital cortex, or pons. A 2-fold greater extent of neurodegeneration was obtained in ventral portions of the olfactory bulb, entorhinal cortex, temporal cortex, and dentate gyrus compared with the dorsal portions of these structures. The pattern of increase in (3H)PK 11195 binding closely matched the patterns of neuronal degeneration reported following parenteral kainate injection. These findings strengthen the notion that quantitative autoradiography of (3H)PK 11195 is a valuable tool to quantify the extent of neuronal degeneration.

  15. Experimental studies on tissue distribution of /sup 14/C-labelled antimicrobial agent in otorhinolaryngological field. Application of macro-autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Murai, K.; Baba, S. (Nagoya City Univ. (Japan). Faculty of Medicine); Morita, S.; Ishigami, M.


    The tissue distribution of nalidixic acid analog OPC-7241 was investigated. Autoradiography was performed 30 minutes after intravenous administration of /sup 14/C-OPC-7241 in 2.96 MBq(80.0 mg/kg to New Zealand White rabbit. Radiometry was performed 30 minutes, 1 hour and 3 hours after intravenous administration of /sup 14/C-OPC-7241 in 740 kBq(20.0 mg/kg to NZW rabbits. Such care was taken in filling the carboxymethyl cellulose paste into the paranasal cavity, nasal cavity, oral cavity and external ear canal not to damage mechanically. Horizontal frozen sections parallel to mandibular basis were cut in 50 thickness in a cryostat. Blackening of /sup 14/C-OPC-7241 was most significant in soft palate and its expanse which is apparently the same gland-like tissues as soft palate when stained with hematoxylin and eosin. Significant radioactivity was recognized in tonsils, the cartilage of external ear canal, septal cartilage, periodontal membrane, dental pulp, muscle and concha nasalis ventralis. The levels were low in ethmoid cells, mucosa of tympanic cavity, cochlea, maxillary sinus and bone. Quantitatively, the levels of /sup 14/C-OPC-7241 radioactivity were high in soft palate, mandibular gland and tonsils. Radioactivity was significant in tongue and concha nasalis ventralis. The levels were low in turbinates, ethmoid cells, maxillary sinus, septal cartilage, septal mucosa, cochlea, brain, optic nerve and lens. Thus radiometric results agreed with the above autoradiographic findings. Macro-autoradiography can be one of useful means for the evaluation chemotherapeutic agents for possible clinical application.

  16. Quantitative Single-Particle Digital Autoradiography with α-Particle Emitters for Targeted Radionuclide Therapy using the iQID Camera

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Brian W.; Frost, Sophia; Frayo, Shani; Kenoyer, Aimee L.; Santos, E. B.; Jones, Jon C.; Green, Damian J.; Hamlin, Donald K.; Wilbur, D. Scott; Fisher, Darrell R.; Orozco, Johnnie J.; Press, Oliver W.; Pagel, John M.; Sandmaier, B. M.


    Abstract Alpha emitting radionuclides exhibit a potential advantage for cancer treatments because they release large amounts of ionizing energy over a few cell diameters (50–80 μm) causing localized, irreparable double-strand DNA breaks that lead to cell death. Radioimmunotherapy (RIT) approaches using monoclonal antibodies labeled with alpha emitters may inactivate targeted cells with minimal radiation damage to surrounding tissues. For accurate dosimetry in alpha-RIT, tools are needed to visualize and quantify the radioactivity distribution and absorbed dose to targeted and non-targeted cells, especially for organs and tumors with heterogeneous radionuclide distributions. The aim of this study was to evaluate and characterize a novel single-particle digital autoradiography imager, iQID (ionizing-radiation Quantum Imaging Detector), for use in alpha-RIT experiments. Methods: The iQID camera is a scintillator-based radiation detection technology that images and identifies charged-particle and gamma-ray/X-ray emissions spatially and temporally on an event-by-event basis. It employs recent advances in CCD/CMOS cameras and computing hardware for real-time imaging and activity quantification of tissue sections, approaching cellular resolutions. In this work, we evaluated this system’s characteristics for alpha particle imaging including measurements of spatial resolution and background count rates at various detector configurations and quantification of activity distributions. The technique was assessed for quantitative imaging of astatine-211 (211At) activity distributions in cryosections of murine and canine tissue samples. Results: The highest spatial resolution was measured at ~20 μm full width at half maximum (FWHM) and the alpha particle background was measured at a rate of (2.6 ± 0.5) × 10–4 cpm/cm2 (40 mm diameter detector area). Simultaneous imaging of multiple tissue sections was performed using a large-area iQID configuration (ø 11.5 cm

  17. Quantitative single-particle digital autoradiography with α-particle emitters for targeted radionuclide therapy using the iQID camera

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Brian W., E-mail: [Pacific Northwest National Laboratory, Richland, Washington 99354 and College of Optical Sciences, The University of Arizona, Tucson, Arizona 85719 (United States); Frost, Sofia H. L.; Frayo, Shani L.; Kenoyer, Aimee L.; Santos, Erlinda; Jones, Jon C.; Orozco, Johnnie J. [Fred Hutchinson Cancer Research Center, Seattle, Washington 98109 (United States); Green, Damian J.; Press, Oliver W.; Pagel, John M.; Sandmaier, Brenda M. [Fred Hutchinson Cancer Research Center, Seattle, Washington 98109 and Department of Medicine, University of Washington, Seattle, Washington 98195 (United States); Hamlin, Donald K.; Wilbur, D. Scott [Department of Radiation Oncology, University of Washington, Seattle, Washington 98195 (United States); Fisher, Darrell R. [Dade Moeller Health Group, Richland, Washington 99354 (United States)


    Purpose: Alpha-emitting radionuclides exhibit a potential advantage for cancer treatments because they release large amounts of ionizing energy over a few cell diameters (50–80 μm), causing localized, irreparable double-strand DNA breaks that lead to cell death. Radioimmunotherapy (RIT) approaches using monoclonal antibodies labeled with α emitters may thus inactivate targeted cells with minimal radiation damage to surrounding tissues. Tools are needed to visualize and quantify the radioactivity distribution and absorbed doses to targeted and nontargeted cells for accurate dosimetry of all treatment regimens utilizing α particles, including RIT and others (e.g., Ra-223), especially for organs and tumors with heterogeneous radionuclide distributions. The aim of this study was to evaluate and characterize a novel single-particle digital autoradiography imager, the ionizing-radiation quantum imaging detector (iQID) camera, for use in α-RIT experiments. Methods: The iQID camera is a scintillator-based radiation detection system that images and identifies charged-particle and gamma-ray/x-ray emissions spatially and temporally on an event-by-event basis. It employs CCD-CMOS cameras and high-performance computing hardware for real-time imaging and activity quantification of tissue sections, approaching cellular resolutions. In this work, the authors evaluated its characteristics for α-particle imaging, including measurements of intrinsic detector spatial resolutions and background count rates at various detector configurations and quantification of activity distributions. The technique was assessed for quantitative imaging of astatine-211 ({sup 211}At) activity distributions in cryosections of murine and canine tissue samples. Results: The highest spatial resolution was measured at ∼20 μm full width at half maximum and the α-particle background was measured at a rate as low as (2.6 ± 0.5) × 10{sup −4} cpm/cm{sup 2} (40 mm diameter detector area

  18. The centuries-old and thousand- year oscillations of uranium distribution in the Lake Baikal sediments, according to the neutron-fission (n,f)-autoradiography (United States)

    Kirichenko, Ivan; Zhmodik, Sergey; Belyanin, Dmitriy; Khlistov, Oleg


    The trace elements local distribution data, particularly (U, P, Br, Mo, BiSi et. all) in a lake and oceans bottom sediments reflects the conditions of those sediments formation, and correlates with changes in paleoclimatic conditions. In papers [Colman et all, 1995; Goldberg et all, 2000, etc.] established that the concentrations of some elements contained in the bottom sediments of Lake Baikal, in particular BiSi, Sr / Ba, Sr / Rb, Ti, U et al., reflect changes in insolation caused by periodic oscillations parameters Earth's orbit (Milankovitch cycles). At the same time, a bottom sediments of the largest continental lake (Lake Baikal), can keep a record of changes less periodicity. Our research focuses on the study of the spatial distribution of uranium with high resolution in the bottom sediments of Lake Baikal. The purpose of this research is determination the centure-old and thousand- old year oscillations in the concentration of uranium in the sediments of Lake Baikal. Fragments of the lake sediment columns taken from the axial part of the Akademicheskiy Ridge in Lake Baikal (stations coordinates St -8 (53 32'15"N 107 56'25"E); - and St11 - (53 33'51"N 108 00'05"E) were studied using complex of local analysis methods, such as: n, f - and n, β-autoradiography, SEM. The distributions of uranium and phosphorus in the authigenic component of sediments along the whole columns length (with the resolution of 10 micron which corresponds to the time resolution of about six months) have been studied by the autoradiography method. Statistical data analysis (Fourier and wavelet analysis) were used for detection oscillations in the uranium concentration Three main different factors of concentrators were established for uranium and phosphorus in the sediments of the Academic mountain range:1) sedimentation, 2) nutrient,3) diagenetic. The periodicity (range from 100 to 1,000 years), in the distribution of authigenic uranium in the sediment column were identified by

  19. Changes in mACh, NMDA and GABA(A) receptor binding after lateral fluid-percussion injury: in vitro autoradiography of rat brain frozen sections. (United States)

    Sihver, S; Marklund, N; Hillered, L; Långström, B; Watanabe, Y; Bergström, M


    Adult rats were subjected to a moderate lateral fluid percussion injury (FPI), followed by survival periods of 2 and 12 h. Regional NMDA subtype glutamate, muscarinic acetylcholine and GABA(A) receptor binding in various brain regions was analysed by quantitative in vitro autoradiography and short-lived positron emission tomography tracers [11C]cyano-dizocilpine, 4-N-[11C]methylpiperidylbenzilate (4-N-[11C]MPB), and [11C]flumazenil, respectively. The binding potential (BP, Bmax/KD) was calculated. The data with [11C]cyano-dizocilpine showed a significant decrease in BP bilaterally for the frontoparietal cortex and hippocampus at both time points, in comparison with that of the sham-operated controls. At 12 h the decrease was significantly more prominent for the ipsilateral cortex and hippocampus than for the contralateral side. The BP of 4-N-[11C]MPB was significantly decreased after 2 h for the trauma-side hippocampus, and after 12 h it had decreased for the trauma-site cortex and the bilateral hippocampus. The [11C]flumazenil exhibited a significant decrease in BP for the trauma-site cortex and the underlying hippocampus by 2 h after the traumatic brain injury. After 12 h a significantly decreased BP was observed only for the trauma-site cortex. The finding of a decreased BP demonstrates the involvement of these receptor systems in the development of cellular dysfunction, which is widespread and not limited to the site of lateral FPI.

  20. Double labelling of tissue combining tritiated thymidine autoradiography with immunodetection of bromodeoxyuridine: the autoradiographic significance of inhibition of thymidine incorporation into DNA by bromodeoxyuridine given simultaneously

    Energy Technology Data Exchange (ETDEWEB)

    Hume, W.J.; Thompson, J. (Leeds Univ. (UK). School of Dentistry)


    The authors describe a method for combining tritiated thymidine (({sup 3}H)TdR) autoradiography with immunoperoxidase detection of bromodeoxyuridine (BrdU) in paraffin-embedded tissues, which was used to examine, in mouse tongue epithelium, the inhibition of incorporation into DNA of ({sup 3}H)TdR by simultaneous injection of BrdU in the doses that both compounds are likely to be used in cell proliferation studies. The inhibition of uptake into DNA of ({sup 3}H)TdR from 0.23 to 1.85 MBq (6.25 to 50 {mu}Ci) per animal, produced by a simultaneous injection of 2.5 mg BrdU shows a linear, dose-dependent relationship. Provided the injected dose (in {mu}Ci per animal) multiplied by the autoradiographic exposure time (in days) is greater than a value of 700, then all cells that are labelled after incorporation of ({sup 3}H)TdR alone are also labelled after simultaneous double labelling, despite the latter producing a lower average grain count. (author).

  1. Fluorescence histochemistry and autoradiography of adrenergic nerves in the renal juxtaglomerular complex of mammals and man, with special regard to the efferent arteriole. (United States)

    Dolezel, S; Edvinsson, L; Owman, C; Owman, T


    The adrenergic innervation of the juxtaglomerular complex was studied in kidneys from mice, rats, guinea-pigs, rabbits, cats, dogs, pigs, monkeys, and humans using fluorescence histochemistry of neuronal noradrenaline and autoradiography of 3H-noradrenaline. The localization of the nerves was established by phase contrast optics or by perfusing the vascular system with India ink. Adrenergic nerve terminals, exhibiting a formaldehyde-induced fluorescence and having the ability to take up and accumulate 3H-noradrenaline, were easily identified when they enclosed the glomerular afferent arteriole. They continued in between and close to the macula densa and lacis cells to supply the glomerular efferent arteriole. The nerves could be seen to accompany this arteriole for a considerable distance until they branched off to the vasa recta in the juxtamedullary region and to adjacent cortical veins. This innervation pattern was found to be a constant feature except in kidneys from guinea-pigs and cats, in which post-glomerular units. The fluorescence in all adrenergic fibres supplying the juxtaglomerular complex disappeared after removal of the aortico-renal ganglion, showing that they belong to a common system of renal sympathetic nerves.

  2. Studies on the percutaneous absorption of /sup 14/C-labelled Flurbiprofen, 3. Whole body autoradiography of rats and guinea-pigs

    Energy Technology Data Exchange (ETDEWEB)

    Nagao, Soshichi; Sakai, Takeo; Hayakawa, Toru (Nihon Univ., Tokyo. Coll. of Agriculture and Veterinary Medicine)


    Whole body autoradiography was carried out to clarify and compare the distribution of /sup 14/C-labelled Flurbiprofen which was applied to the skin as an ointment in rats and guinea-pigs. Both in rats and guinea-pigs almost the same autoradiogram was gained. The radioactivity was strongest at the skin area inspite of the time elapse, showing that the drug was fixed in the site of skin applied. In other parts of the body, however, it was small except the kidney and intestine. It seemed that the absorption of the drug was a little although the migration of the drug into the blood circulation is fast at the beginning as was shown in pigs previously. A stronger radioactivity in the kidney and intestine might indicate that a main pathway of excretion of this drug was through those two organs. Absorption, distribution and excretion of the drug were not different between rats and guinea-pigs, similar to those observed in pigs.

  3. Syntheses of [sup 14]C-ochratoxin A and [sup 14]C-ochratoxin B and a comparative study of their distribution in rats using whole body autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Breitholtz-Emanuelsson, A.; Hult, K. (Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm (Sweden)); Fuchs, R. (Department of Toxicology, Institute for Medical Research and Occupational Health, University of Zagreb, Zagreb (Yugoslavia)); Appelgren, L.-E. (Department of Pharmacology and Toxicology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala (Sweden))


    Methods for preparation of labelled ochratoxin A and B are described. The method for preparation of labelled ochratoxin B involves the synthesis of the azide of ochratoxin [beta] via the mixed anhydride and subsequent conjugation to labelled phenylalanine to yield [sup 14]C-ochratoxin B. The labelled ochratoxins were injected into male Wistar rats and after different survival times they were sacrificed and subjected to whole body autoradiography. The distribution pattern of ochratoxin A in the rat did not differ from that earlier registered for mouse. The previously known, high susceptibility of rats (and not mice) to ochratoxin A-induced cancer could thus not be explained by an accumulation of the toxin in specific cells or organs. The distribution patterns of ochratoxin A and B were almost congruent - the only apparent difference being a much longer retention of the labelled ochratoxin A in the blood compared to ochratoxin B, which was much faster excreted. When analyzing tissue extract for labelled metabolites only the extracts from the rats injected with ochratoxin B were found to contain easily detectable concentrations, while no metabolites of ochratoxin A were seen. (au).

  4. Comparison of Drug Distribution Images from Thin Tissue Sections Obtained Using Desorption Electrospray Ionization Tandem Mass Spectrometry and Whole-Body Autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Kertesz, Vilmos [ORNL; Van Berkel, Gary J [ORNL; Vavek, Marissa [Merck Research Laboratories; Koeplinger, Kenneth A. [Merck Research Laboratories; Schneider, Bradley B [MDS Sciex; Covey, Thomas R. [MDS Sciex


    Desorption electrospray ionization tandem mass spectrometry (DESI-MS/MS) and whole-body autoradiography (WBA) were used for chemical imaging of whole-body thin tissue sections of mice intravenously dosed with propranolol (7.5 mg/kg). DESI-MS/MS imaging utilized selected reaction monitoring detection performed on an AB/MDS SCIEX 4000 QTRAP mass spectrometer equipped with a prototype extended length particle discriminator interface. Propranolol images of the tissue sections using DESI-MS/MS were obtained at surface scan rates of 0.1, 0.5, 2 and 7 mm/s. Although signal decreased with increasing scan rate, useful whole-body images for propranolol were obtained from the tissues even at 7 mm/s, which required just 79 min of analysis time. Attempts to detect and image the distribution of the known propranolol metabolites were unsuccessful. Regions of the tissue sections showing the most radioactivity from WBA sections were excised and analyzed by HPLC with radiochemical detection to determine relative levels of propranolol and metabolites present. Comparison of the DESI-MS/MS signal for propranolol and the radioactivity attributed to propranolol from WBA sections indicated nominal agreement between the two techniques for the amount of propranolol in the brain, lung, and liver. Data from the kidney showed an unexplained disparity between the two techniques. The results of this study show the feasibility of using DESI-MS/MS to obtain useful chemical images of a drug in whole-body thin tissue sections following drug administration at a pharmacologically relevant level. Further optimization to improve sensitivity and enable detection of the drug metabolites will be among the requirements necessary to move DESI-MS/MS chemical imaging forward as a practical tool in drug discovery.

  5. Neurogenesis in the vomeronasal epithelium of adult garter snakes: 3. Use of /sup 3/H-thymidine autoradiography to trace the genesis and migration of bipolar neurons

    Energy Technology Data Exchange (ETDEWEB)

    Wang, R.T.; Halpern, M.


    Use of 3H-thymidine autoradiography and unilateral vomeronasal (VN) axotomy has permitted us to demonstrate directly the existence of VN stem cells in the adult garter snake and to trace continuous bipolar neuron development and migration in the normal VN and deafferentated VN epithelium in the same animal. The vomeronasal epithelium and olfactory epithelium of adult garter snakes are both capable of incorporating 3H-thymidine. In the sensory epithelium of the vomeronasal organ, 3H-thymidine-labeled cells were initially restricted to the base of the undifferentiated cell layer in animals surviving 1 day following 3H-thymidine injection. With increasing survival time, labeled cells progressively migrated vertically within the receptor cell column toward the apex of the bipolar neuron layer. In both the normal and denervated VN epithelium, labeled cells were observed through the 56 days of postoperative survival. In the normal epithelium, labeled cells were always located within the matrix of the intact receptor cell columns. However, labeled cells of the denervated epithelium were always located at the apical front of the newly formed cell mass following depletion of the original neuronal cell population. In addition, at postoperative days 28 and 56, labeled cells of the denervated VN epithelium achieved neuronal differentiation and maturation by migrating much farther away from the base of the receptor cell column than the labeled cells on the normal, unoperated contralateral side. This study directly demonstrates that basal cells initially incorporating 3H-thymidine are indeed stem cells of the VN epithelium in adult garter snakes.

  6. Estradiol receptor binding to the epithelium of uterine lumen and glands: region- and time-related changes during preimplantation and periimplantation periods studied by autoradiography. (United States)

    Zorn, Telma M T; Soto-Suazo, Mauricio; Pellegrini, Cleusa R; Oliveira, José G; Stumpf, Walter E


    The presence and changes of estradiol nuclear binding and related functions in uterine luminal and glandular epithelium were studied before and after blastocyst implantation using receptor autoradiography with (3)H-estradiol-17beta in association with (3)H-thymidine incorporation and immunocytochemical binding of antibody to estrogen receptor ER-alpha. (3)H-estradiol nuclear binding is present but variable during days 1.5-7.5 of pregnancy. Sites of strong nuclear binding of (3)H-estradiol exhibit strong immunocytochemical staining with ER-alpha antibody. Qualitative and quantitative evaluation of autoradiograms reveal that there is a general increase of nuclear (3)H-estradiol binding during the first 3 days after fertilization in both luminal and glandular epithelium. The binding of estradiol is stronger in glandular epithelium from day 2.5 to day 7.5, paralleled by a rise in (3)H-thymidine incorporation on day 2.5. By comparison, in the epithelium of the uterine lumen (3)H-estradiol nuclear binding is low, but relatively high in epithelial cells at lateral branching of the lumen where the increase in (3)H-estradiol binding corresponds to an increased labeling index with (3)H-thymidine. A highly differentiated binding of (3)H-estradiol to luminal and glandular epithelium was demonstrated with region- and time-specific changes of related effects on cell proliferation, differentiation, and secretion, probably involving involution and remodeling. The strong (3)H-estradiol binding to glandular epithelium suggests that estradiol exerts pronounced effects on glandular activities in the periimplantation period.

  7. Effect of irradiation on the formation of secretory granules in the mouse submandibular glands; Study of electron microscopic autoradiography with sup 3 H-leucine

    Energy Technology Data Exchange (ETDEWEB)

    Kaito, Tomoi; Nasu, Masanori; Kondoh, Nobuyoshi; Furumoto, Keiichi (Nippon Dental Univ., Tokyo (Japan))


    To determine the effect of radiation on secretory granule morphosis of the submandibular acinar cells, the lower jaw of the rat irradiated with 10 Gy of X-ray. We observed intracellular localization of reducing argentum grains three days after irradiation by electron microscopic autoradiography with {sup 3}H-Leucine (RI). The argentum grain ratio localizing in the ganular endoplasmic reticulum stood at the same level both in the irradiated group (IG) and in the non-irradiated group (NG) 30 minutes after injection of RI. But in NG the ratio declined sharply at the 60-minute mark. One hundred and eighty minutes after the injection of RI, the argentum grain ratio in NG showed the same level at the 60-minute mark. While in IG the ratio was lower than that at the 60 minutes after the injection but not as low as in NG. The argentum grain ratio localized in the Golgi complex showed the same level in both NG and IG and decreased with time after the injection of RI. The argentum grain ratio localized in the secretory granule increased with time after the injection of RI in both NG and IG. The ratio in IG was entirely lower than that in NG. A retarding trend of secretory progress was recognized, showing a high ratio even 180 minutes after injection in IG, while the ratio reached a peak of 60 minutes in NG after the injection of RI. The argentum grain localizing in the other cellular organelles which have no relation with secretory granule morphosis showed a ratio below 10 percent in both NG and IG at each testing time after the injection of RI and no distinctive change was recognized in the procedure. The secretory granule morphosis of the mouse's submandibule was clarified to be retarded three days after 10 Gy irradiation. (J.P.N.).

  8. The effect of irradiation on the intracellular transportation of the parotid gland acinar cells in the mouse. Localization of monosaccharides studied by electron microscopic autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Matsunaga, Hajime (Nippon Dental Univ., Tokyo (Japan))


    The present study was designed to investigate the effects of radiation on the ability to ingest monosaccharides and intracellular transportation in the parotid gland in mice. The submandibular regions, including the parotid gland, was exposed to 10 Gy of X-rays. Three days after irradiation, the localization of reducing silver grains in organelles was determined, using electron microscopic autoradiography with H-3 labeled galactosamine, glucosamine, fucose, and mannose. In the non-irradiated group, the proportion of reducing silver grains in the acinar cells began to increase 15 min after administration of monosaccharides, reached a peak at 180 min, and thereafter decreased. Similar findings were observed in the irradiated group, although the values were lower than the non-irradiated group. The proportion of reducing silver grains in the endoplasmic reticulum reached a peak at 15 min in both the non-irradiated and irradiated groups, and gradually decreased until 120 min. Thereafter, it became almost constant and low, but the proportion in the irradiated group was slightly higher than in the non-irradiated group. The proportion of reducing silver grains in the Golgi apparatus was maximum at 60 min in the non-irradiated group, and gradually decreased until 360 min. A similar tendency was seen in the irradiated group, although its variation was not so marked as in the non-irradiated group. The proportion of reducing silver grains in the condensing vacuoles was maximum at 120 min, and thereafter, it decreased; the decrease was only slight in the irradiated group. The proportion of reducing silver grains in secretory granules increased with time in both the non-irradiated and irradiated groups, although this was only slight in the irradiated group, and reached a peak at 360 min. Transportation of monosaccharides in an acinar cell was found to be delayed by irradiation. (N.K.).

  9. Evaluation of cardiac adrenergic neuronal damage in rats with doxorubicin-induced cardiomyopathy using iodine-131 MIBG autoradiography and PGP 9.5 immunohistochemistry

    Energy Technology Data Exchange (ETDEWEB)

    Jeon, T.J.; Lee, J.D. [Division of Nuclear Medicine, Department of Diagnostic Radiology, Yonsei University Medical College, Seoul (Korea); Research Institute of Radiological Science, Yonsei University Medical College, Seoul (Korea); Ha, J.-W. [Cardiology Division, Yonsei Cardiovascular Center, Yonsei University Medical College, Seoul (Korea); Yang, W.I.; Cho, S.H. [Department of Pathology, Yonsei University Medical College, Seoul (Korea)


    Doxorubicin is one of the most useful anticancer agents, but its repeated administration can induce irreversible cardiomyopathy as a major complication. The purpose of this study was to investigate doxorubicin toxicity on cardiac sympathetic neurons using iodine-131-metaiodobenzylguanidine (MIBG) and protein gene product (PGP) 9.5 immunohistochemistry, which is a marker of cardiac innervation. Wistar rats were treated with doxorubicin (2 mg/kg, i.v.) once a week for 4 (n=5), 6 (n=6) or 8 (n=7) weeks consecutively. Left ventricular ejection fraction (LVEF), calculated by M-mode echocardiography, was used as an indicator of cardiac function. Plasma noradrenaline (NA) concentration was measured by high-performance liquid chromatography (HPLC). {sup 131}I-MIBG uptake of the left ventricular wall (24 ROIs) was measured by autoradiography. {sup 131}I-MIBG uptake pattern was compared with histopathological results, the neuronal population on PGP 9.5 immunohistochemistry and the degree of myocyte damage assessed using a visual scoring system on haematoxylin and eosin and Masson's trichrome staining. LVEF was significantly decreased in the 8-week group (P<0.05). The serum NA level also showed no statistical difference until 4 weeks and was significantly increased in the 8-week group (P<0.05). MIBG uptake was decreased in the 6- and 8-week groups (P<0.05), and was closely correlated with the reduction in the number of nerve fibres on PGP 9.5 stain. Myocyte damage was seen only in the 8-week group. Neuronal population and the {sup 131}I-MIBG uptake ratio of subepicardium to subendocardium were significantly increased (P<0.05) in the 8-week group as compared with the control group. It may be concluded that radioiodinated MIBG is a reliable marker for the detection of cardiac adrenergic neuronal damage in doxorubicin-induced cardiomyopathy; it detects such damage earlier than do other clinical parameters and in this study showed a good correlation with the reduction in the

  10. Effect of irradiation for intracellular transport on mouse parotid gland; Study of electron microscopic autoradiography with [sup 3]H-leucine

    Energy Technology Data Exchange (ETDEWEB)

    Kondou, Nobuyoshi (Nippon Dental Univ., Tokyo (Japan))


    Using light and electron microscopic autoradiographies by means of [sup 3]H-leucine, the influence of X-radiation, 10 Gy upon the submandibular region including parotid gland of a mouse was examined. The number of reduced silver grain per unit area of acinar cell was compared, and the rate of reduced silver grain localized in the intracellular organelle involved in the synthesis and transport of protein was observed. In the non-radiation (NR) group, reduced silver grain in the acinar cell of parotid gland showed the maximum value 30 minutes after [sup 3]H-leucine administration and thereafter decreased with time. Even the 3 and 14 post-radiation (PR) day-groups showed the maximum values at 30 minutes, but to a lesser extent than the NR groups, and subsequent time-course was noted a little. Reduced silver grain localized in the rough surfaced reticulum showed the highest rate at 15 minutes for the NR, 3 and 14 PR groups, and thereafter decreased abruptly. In comparing the rate of reduced silver grain localized in Golgi apparatus, the NR group showed the highest rate at 60 minutes and gradually decreased thereafter. The 3 PR group showed the highest rate at 60 minutes and similar tendency up to 120 minutes. The 14 PR group showed almost the similar tendency to the NR group. Reduced silver particles localized peri- and intra-secretory granules showed higher rate at 60 minutes for the NR group. In the 3 PR group, peri- and intra-secretory granules showed almost the same rate at 180 minutes, with a time lag for the transition of [sup 3]H-leucine to the secretory granules. In the 3 and 14 PR groups, similar order of rate was noted at 60 minutes between peri- and intra-secretory granules, with a transition time approximating to that of the NR group. Subsequent discharge, however, showed a delay tendency. Pathohistological examination revealed strong morphological changes of intracellular organelle in the 3 PR group and less remarkable changes in the 14 PR group. (author).

  11. High resolution beta imaging systems for quantitative autoradiography. Potential applications in the field of drug or radiopharmaceuticals developments; Autoradiographie quantitative a l'aide des nouveaux systemes d'imagerie Beta a haute resolution: applications a l'evaluation de medicaments ou a la mise au point de radiopharmaceutiques

    Energy Technology Data Exchange (ETDEWEB)

    Barthe, N


    Autoradiography, an imaging technique allowing to get the spatial distribution of a radioactive substance in a tissue, is generally obtained by means of films or photographic emulsions. This is a difficult methodology with several limitations: the major one being quantification which is not precise. We have contributed to the development of new high resolution beta imaging systems to replace the photographic emulsion by a direct counting of beta radioactivity. They have similar or improved performances compared to the film. The sensitivity of the detection is specially enhanced, multiplying by a factor of 100 to 500 the rapidity of image acquisition. Linearity of the detectors is improved giving a better dynamic range than the film. So quantification of radioactivity in the tissue sections is obtained with a good reliability. Multi-isotopes imaging is also achieved by using algorithms of separation based on the different characteristics of energies or decaying schemes of the isotopes. Many potential applications can be found in the development of drugs or radiopharmaceuticals. (author)

  12. Direct comparison of local cerebral blood flow rates measured by MRI arterial spin-tagging and quantitative autoradiography in a rat model of experimental cerebral ischemia. (United States)

    Ewing, James R; Wei, Ling; Knight, Robert A; Pawa, Swati; Nagaraja, Tavarekere N; Brusca, Thomas; Divine, George W; Fenstermacher, Joseph D


    The present study determined cerebral blood flow (CBF) in the rat using two different magnetic resonance imaging (MRI) arterial spin-tagging (AST) methods and 14C-iodoantipyrine (IAP)-quantitative autoradiography (QAR), a standard but terminal technique used for imaging and quantitating CBF, and compared the resulting data sets to assess the precision and accuracy of the different techniques. Two hours after cerebral ischemia was produced in eight rats via permanent occlusion of one middle cerebral artery (MCA) with an intraluminal suture, MRI-CBF was measured over a 2.0-mm coronal slice using single-coil AST, and tissue magnetization was assessed by either a spin-echo (SE) or a variable tip-angle gradient-echo (VTA-GE) readout. Subsequently ( approximately 2.5 hours after MCA occlusion), CBF was assayed by QAR with the blood flow indicator 14C-IAP, which produced coronal images of local flow rates every 0.4 mm along the rostral-caudal axis. The IAP-QAR images that spanned the 2-mm MRI slice were selected, and regional flow rates (i.e., local CBF [lCBF]) were measured and averaged across this set of images by both the traditional approach, which involved reader interaction and avoidance of sectioning artifacts, and a whole film-scanning technique, which approximated total radioactivity in the entire MRI slice with minimal user bias. After alignment and coregistration, the concordance of the CBF rates generated by the two QAR approaches and the two AST methods was examined for nine regions of interest in each hemisphere. The QAR-lCBF rates were higher with the traditional method of assaying tissue radioactivity than with the MRI-analog approach; although the two sets of rates were highly correlated, the scatter was broad. The flow rates obtained with the whole film-scanning technique were chosen for subsequent comparisons to MRI-CBF results because of the similarity in tissue "sampling" among these three methods. As predicted by previous modeling, "true" flow rates

  13. Syntheses and in vitro evaluation of fluorinated naphthoxazines as dopamine D2/D3 receptor agonists: radiosynthesis, ex vivo biodistribution and autoradiography of [{sup 18}F]F-PHNO

    Energy Technology Data Exchange (ETDEWEB)

    Vasdev, Neil [PET Centre for Addiction and Mental Health, Toronto, Ontario, Canada, M5T-1R8 (Canada) and Department of Psychiatry, University of Toronto, Toronto, Ontario, M5T-1R8 (Canada)]. E-mail:; Seeman, Philip [Department of Psychiatry, University of Toronto, Toronto, Ontario, M5T-1R8 (Canada); Department of Pharmacology, University of Toronto, Toronto, Ontario, M5S-1A8 (Canada); Garcia, Armando [PET Centre for Addiction and Mental Health, Toronto, Ontario, M5T-1R8 (Canada); Stableford, Winston T. [PET Centre for Addiction and Mental Health, Toronto, Ontario, M5T-1R8 (Canada); Nobrega, Jose N. [PET Centre for Addiction and Mental Health, Toronto, Ontario, M5T-1R8 (Canada); Department of Psychiatry, University of Toronto, Toronto, Ontario, M5T-1R8 (Canada); Department of Pharmacology, University of Toronto, Toronto, Ontario, M5S-1A8 (Canada); Houle, Sylvain [PET Centre for Addiction and Mental Health, Toronto, Ontario, M5T-1R8 (Canada); Department of Psychiatry, University of Toronto, Toronto, Ontario, M5T-1R8 (Canada); Wilson, Alan A. [PET Centre for Addiction and Mental Health, Toronto, Ontario, M5T-1R8 (Canada); Department of Psychiatry, University of Toronto, Toronto, Ontario, M5T-1R8 (Canada)


    Introduction: Carbon-11-labeled (+)-4-propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol ([{sup 11}C]-(+)-PHNO) is a dopamine D2/D3 agonist radioligand that is currently used to image the high-affinity state of dopamine receptors in humans with positron emission tomography (PET). The present study reports the preparation and evaluation of fluorinated (+)-PHNO derivatives. Methods: Five fluorinated (+)-PHNO derivatives were synthesized and tested in vitro for inhibition of binding of [{sup 3}H]domperidone in homogenates of rat striatum and inhibition of binding to [{sup 3}H]-(+)-PHNO in homogenates of human-cloned D2Long receptors in Chinese hamster ovary cells and rat striatum. Radiolabeling with fluorine-18 was carried out for the most promising candidate, N-fluoropropyl-(+)-HNO (F-PHNO), and ex vivo biodistribution and autoradiography studies with this radiopharmaceutical were performed in rodents. Results: (+)-PHNO and the fluorinated analogs inhibited binding of [{sup 3}H]domperidone and [{sup 3}H]-(+)-PHNO to the high- and low-affinity states of dopamine D2 receptors, consistent with D2 agonist behavior. The average dissociation constant at the high-affinity state of D2, K {sub i} {sup High}, was 0.4 nM for F-PHNO and proved to be equipotent with (+)-PHNO (0.7 nM). All other fluorinated derivatives were significantly less potent (K {sub i} {sup High}=2-102 nM). The most promising candidate, F-PHNO, was labeled with fluorine-18 in 5% uncorrected radiochemical yield, with respect to starting fluoride. Ex vivo biodistribution and autoradiography studies in rodents revealed that [{sup 18}F]F-PHNO rapidly enters the rodent brain. However, this radiotracer does not reveal specific binding in the brain and is rapidly cleared. Conclusions: Five novel dopamine D2/D3 agonists based on (+)-PHNO were synthesized and evaluated in vitro. F-PHNO was shown to behave as a potent D2 agonist in vitro and was therefore radiolabeled with fluorine-18. Despite the

  14. Application of neutron capture autoradiography to Boron Delivery seeking techniques for selective accumulation of boron compounds to tumor with intra-arterial administration of boron entrapped water-in-oil-in-water emulsion

    Energy Technology Data Exchange (ETDEWEB)

    Mikado, S. [Physical Science Laboratories, College of Industrial Technology, Nihon University, Chiba (Japan)], E-mail:; Yanagie, H. [Department of Nuclear Engineering and Management, University of Tokyo, Tokyo (Japan); Cooperative Unit of Medicine and Engineering, University of Tokyo Hospital, Tokyo (Japan); Yasuda, N. [Fundamental Technology Center, National Institute of Radiological Sciences, Chiba (Japan); Higashi, S.; Ikushima, I. [Miyakonojyo Metropolitan Hospital, Miyazaki (Japan); Mizumachi, R.; Murata, Y. [Department of Pharmacology, Kumamoto Institute Branch, Mitsubishi Chemical Safety Institute Ltd., Kumamoto (Japan); Morishita, Y. [Department of Human and Molecular Pathology, University of Tokyo, Tokyo (Japan); Nishimura, R. [Faculty of Agriculture, Laboratory of Veterinary Surgery, University of Tokyo (Japan); Shinohara, A. [Department of Humanities, The Graduate School of Seisen University, Tokyo (Japan); Ogura, K. [Physical Science Laboratories, College of Industrial Technology, Nihon University, Chiba (Japan); Sugiyama, H. [Cooperative Unit of Medicine and Engineering, University of Tokyo Hospital, Tokyo (Japan); Iikura, H.; Ando, H. [Japan Atomic Energy Agency, Ibaraki (Japan); Ishimoto, M. [Department of Nuclear Professional School, University of Tokyo (Japan); Takamoto, S. [Cooperative Unit of Medicine and Engineering, University of Tokyo Hospital, Tokyo (Japan); Department of Cardiac Surgery, University of Tokyo Hospital, Tokyo (Japan); Eriguchi, M. [Cooperative Unit of Medicine and Engineering, University of Tokyo Hospital, Tokyo (Japan); Department of Microbiology, Syowa University School of Pharmaceutical Sciences, Tokyo (Japan); Takahashi, H. [Department of Nuclear Engineering and Management, University of Tokyo, Tokyo (Japan); Cooperative Unit of Medicine and Engineering, University of Tokyo Hospital, Tokyo (Japan); Kimura, M. [Department of Physics, Toho University, Chiba (Japan)


    It is necessary to accumulate the {sup 10}B atoms selectively to the tumor cells for effective Boron Neutron Capture Therapy (BNCT). In order to achieve an accurate measurement of {sup 10}B accumulations in the biological samples, we employed a technique of neutron capture autoradiography (NCAR) of sliced samples of tumor tissues using CR-39 plastic track detectors. The CR-39 track detectors attached with the biological samples were exposed to thermal neutrons in the thermal column of the JRR3 of Japan Atomic Energy Agency (JAEA). We obtained quantitative NCAR images of the samples for VX-2 tumor in rabbit liver after injection of {sup 10}BSH entrapped water-in-oil-in-water (WOW) emulsion by intra-arterial injection via proper hepatic artery. The {sup 10}B accumulations and distributions in VX-2 tumor and normal liver of rabbit were investigated by means of alpha-track density measurements. In this study, we showed the selective accumulation of {sup 10}B atoms in the VX-2 tumor by intra-arterial injection of {sup 10}B entrapped WOW emulsion until 3 days after injection by using digitized NCAR images (i.e. alpha-track mapping)

  15. Neutron activation autoradiography and scanning macro-XRF of Rembrandt van Rijn's Susanna and the Elders (Gemäldegalerie Berlin): a comparison of two methods for imaging of historical paintings with elemental contrast (United States)

    Alfeld, Matthias; Laurenze-Landsberg, Claudia; Denker, Andrea; Janssens, Koen; Noble, Petria


    Imaging methods with elemental contrast are of great value for the investigation of historical paintings, as they allow for study of sub-surface layers that provide insight into a painting's creation process. Two of the most important methods are neutron activation autoradiography (NAAR) and scanning macro-XRF (MA-XRF). Given the differences between these methods in the fundamental physical phenomena exploited, a theoretical comparison of their capabilities is difficult and until now a critical comparison of their use on the same painting is missing. In this paper, we present a study of Rembrandt van Rijn's painting Susanna and the Elders from the Gemäldegalerie in Berlin employing both techniques. The painting features a considerable number of overpainted features and a wide range of pigments with different elemental tracers, including earth pigments (Mn/Fe), Azurite (Cu), lead white (Pb), vermilion (Hg) and smalt (Co, As). MA-XRF can detect all elements above Si ( Z = 14), suffers from few spectral overlaps and can be performed in a few tens of hours in situ, i.e. in a museum. NAAR requires the stay of the painting at a research facility for several weeks, and inter-element interferences can be difficult to resolve. Also, only a limited number of elements contribute to the acquired autoradiographs, most notably Mn, Cu, As, Co, Hg and P. However, NAAR provides a higher lateral resolution and is less hindered by absorption in covering layers, which makes it the only method capable of visualizing P in lower paint layers.

  16. A comparative autoradiography study in post mortem whole hemisphere human brain slices taken from Alzheimer patients and age-matched controls using two radiolabelled DAA1106 analogues with high affinity to the peripheral benzodiazepine receptor (PBR) system. (United States)

    Gulyás, Balázs; Makkai, Boglárka; Kása, Péter; Gulya, Károly; Bakota, Lidia; Várszegi, Szilvia; Beliczai, Zsuzsa; Andersson, Jan; Csiba, László; Thiele, Andrea; Dyrks, Thomas; Suhara, Tetsua; Suzuki, Kazutoshi; Higuchi, Makato; Halldin, Christer


    The binding of two radiolabelled analogues (N-(5-[125I]Iodo-2-phenoxyphenyl)-N-(2,5-dimethoxybenzyl)acetamide ([125I]desfluoro-DAA1106) and N-(5-[125I]Fluoro-2-phenoxyphenyl)-N-(2-[125I]Iodo-5-methoxybenzyl)acetamide ([125I]desmethoxy-DAA1106) of the peripheral benzodiazepine receptor (PBR) (or TSPO, 18kDa translocator protein) ligand DAA1106 was examined by in vitro autoradiography on human post mortem whole hemisphere brain slices obtained from Alzheimer's disease (AD) patients and age-matched controls. Both [(125)I]desfluoro-IDAA1106 and [(125)I]desmethoxy-IDAA1106 were effectively binding to various brain structures. The binding could be blocked by the unlabelled ligand as well as by other PBR specific ligands. With both radiolabelled compounds, the binding showed regional inhomogeneity and the specific binding values proved to be the highest in the hippocampus, temporal and parietal cortex, the basal ganglia and thalamus in the AD brains. Compared with age-matched control brains, specific binding in several brain structures (temporal and parietal lobes, thalamus and white matter) in Alzheimer brains was significantly higher, indicating that the radioligands can effectively label-activated microglia and the up-regulated PBR/TSPO system in AD. Complementary immunohistochemical studies demonstrated reactive microglia activation in the AD brain tissue and indicated that increased ligand binding coincides with increased regional microglia activation due to neuroinflammation. These investigations yield further support to the PBR/TSPO binding capacity of DAA1106 in human brain tissue, demonstrate the effective usefulness of its radio-iodinated analogues as imaging biomarkers in post mortem human studies, and indicate that its radiolabelled analogues, labelled with short half-time bioisotopes, can serve as prospective in vivo imaging biomarkers of activated microglia and the up-regulated PBR/TSPO system in the human brain.

  17. Examining dopamine D3 receptor occupancy by antipsychotic drugs via [3H]7-OH-DPAT ex vivo autoradiography and its cross-validation via c-fos immunohistochemistry in the rat brain. (United States)

    Davoodi, Nima; te Riele, Paula; Langlois, Xavier


    Dopamine D3 receptors are a major target for drug discovery programs related to psychiatric disorders such as schizophrenia. The ability of a compound to occupy significant levels of D3 receptors is important for achieving therapeutic efficacy in both pre-clinical and clinical settings. Here we attempt to characterise antipsychotic drug-effects at D3 receptors by measuring receptor occupancy via ex-vivo [3H]7-OH-DPAT autoradiography, and further validating this outcome via analysis of Fos-like immunoreactivity (Fos-LI) in the rat major islands of Calleja (ICjM), a brain structure with high D3 expression. Rats were treated subcutaneously with haloperidol (0.04 mg/kg), clozapine (20 mg/kg) and olanzapine (0.63 mg/kg), the selective D2 antagonist L-741626 (2.5 mg/kg) and the selective D3 antagonist SB-277011-A (10 mg/kg). Doses were based on levels of D2 occupancy considered clinically relevant (60-80%). When measuring D3 occupancy, clozapine and SB-277011-A displayed meaningful levels of occupancy (60% and 77%, respectively), haloperidol and olanzapine showed limited occupancy (16% and 27%, respectively), whereas L-741626 showed no occupancy. There were no significant changes in ICjM Fos-LI after L-741626 and haloperidol treatment, minor but significant increases after olanzapine treatment, whereas highly significant increases were seen with SB-277011-A and clozapine. Additionally, pre-treating clozapine with the D1 antagonist SCH23390 caused a significant, albeit non-complete, reduction in Fos-LI, highlighting the D1 agonist property of clozapine. In conclusion, it appears that drugs occupying >50% D3 receptors produce robust increases in ICjM Fos-LI. This study may help to identify the appropriate D3 receptor antagonists that have the potential to be tested in the clinic.

  18. Localization of GABA(B) (R1) receptors in the rat hippocampus by immunocytochemistry and high resolution autoradiography, with specific reference to its localization in identified hippocampal interneuron subpopulations. (United States)

    Sloviter, R S; Ali-Akbarian, L; Elliott, R C; Bowery, B J; Bowery, N G


    Immunocytochemical and autoradiographic methods were used to localize the GABA(B) receptor in the normal rat hippocampus. GABA(B) receptor 1-like immunoreactivity (GBR1-LI) was most intense in presumed GABAergic interneurons of all hippocampal subregions. It was also present throughout the hippocampal neuropil, where it was most intense in the dendritic strata of the dentate gyrus, which are innervated by the perforant pathway and inhibitory dentate hilar cells, and in strata oriens and radiatum of area CA3. The dendritic regions of area CA1 exhibited less GBR1-LI than area CA3. GBR1-LI was detectable in the somata of CA1 pyramidal cells, but was minimal or undetectable within the somata of dentate granule cells and CA3 pyramidal cells. GBR1-LI was similarly minimal in the dentate hilar neuropil, and in stratum lucidum, the two regions that contain granule cell axons and terminals. Nor was GBR1-LI detectable in the inhibitory basket cell fiber systems that surround hippocampal principal cell somata. Fluorescence co-localization studies indicated that significant proportions of interneurons expressing somatostatin, neuropeptide Y, cholecystokinin, calbindin, or calretinin also expressed GBR1-LI constitutively. Conversely, parvalbumin-positive GABAergic basket cells of the dentate gyrus and hippocampus, which form GABA(A) receptor-mediated inhibitory axo-somatic synapses, rarely contained detectable GBR1-LI. High resolution autoradiography with the GABA(B) receptor antagonist CGP 62349 revealed a close correspondence between receptor ligand binding and GBR1-LI, with several notable exceptions. Ligand binding closely matched GBR1-LI throughout the hippocampal, cortical, thalamic, and cerebellar neuropil. However, the hippocampal interneuron somata and dendrites that exhibited the most intense GBR1-LI, and the GBR1-positive somata of CA1 pyramidal cells, did not exhibit a similar density of [3H]-CGP 62349 binding. These data clarify the relationship between

  19. Biomedical applications of digital autoradiography with a MWPC

    Energy Technology Data Exchange (ETDEWEB)

    Bellazzini, R.; Betti, G.; Del Guerra, A.; Massai, M.M.; Ragadini, M.; Spandre, G.; Tonelli, G.; Venturi, R.; Zito, F.


    A Multiwire Proportional Chamber (MWPC) was used as a ..beta../sup -/ radioactivity detector in biological and medical applications. Two different kinds of experiments were performed: the study of variations in the ability of cell clones to incorporate a radioactive precursor of DNA biosynthesis (/sup 3/H-thymidine) and the regional carbohydrate consumption in myocardial tissue by means of a deposit tracer of glucose metabolism (/sup 3/H-deoxyglucose).

  20. Cerebral correlates of depressed behavior in rats, visualized using /sup 14/C-2-deoxyglucose autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Caldecott-Hazard, S.; Mazziotta, J.; Phelps, M.


    /sup 14/C-2-Deoxyglucose (2DG) was used to investigate changes in the rate of cerebral metabolism in 3 rat models of depressed behavior. The models had already been established in the literature and were induced by injections of alpha-methyl-para-tyrosine, withdrawal from chronic amphetamine, or stress. We verified that exploratory behaviors were depressed in each model and that an antidepressant drug, tranylcypromine, prevented the depressed behavior in each model. 2DG studies revealed that the rate of regional glucose metabolism was elevated bilaterally in the lateral habenula of each of the 3 models. Regional metabolic rates were reduced in each model in the dorsal medial prefrontal cortex, anterior ventral nucleus of the thalamus, and inferior colliculus. Forebrain global metabolic rates were also reduced in each of the models. Tranylcypromine prevented the elevated rate of lateral habenula metabolism seen in each of the models alone but did not significantly affect the rates of global metabolism. Our findings of identical metabolic changes in each of the models indicate that these changes are not idiosyncratic to a particular model; rather, they correlate with a generalizable state of depressed exploratory behavior in rats.

  1. Evaluation of the effects of sensory denervation on osteoblasts by 3H-proline autoradiography. (United States)

    Chiego, D J; Singh, I J


    The inferior alveolar nerve was unilaterally resected in 30-day-old mice; other animals were unilaterally sham-operated. At 15, 30, 60, 90, or 150 days after surgery, the mice wee injected with 2 muCi of 3H-proline (sp. act. 1.0Ci/mM) per g of body weight and killed 15, 30, or 60 min later. Autoradiographs were prepared from 5 micron decalcified sagittal sections of mandibles and grain counts made over periosteal osteoblasts mesial to the first molar. In denervated mandibles, osteoblasts incorporated less isotope compared to controls with differences being maximal at the early intervals. These differences became attenuated with time, possibly due to an intrinsic compensatory mechanism, secondary to neurotrophic regulation.

  2. Localization and characterization of renal calcitonin receptors by in vitro autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Sexton, P.M.; Adam, W.R.; Moseley, J.M.; Martin, T.J.; Mendelsohn, F.A.


    Calcitonin receptors were identified in renal cortex and medulla using the radioligand /sup 125/I-salmon calcitonin. Microscopic localization of these receptors revealed binding over medullary and cortical thick ascending limb of the loop of Henle and in distal convoluted tubule. A number of receptor positive cells in the inner medulla were also identified. Characterization of the binding demonstrated a single class of high-affinity binding sites in both the medulla and the cortex with affinity constants of 0.74 +/- 0.09 x 10(9) M-1 and 0.32 +/- 0.05 x 10(9) M-1, respectively, and receptor concentrations of 205 +/- 45 fmol/mg protein and 453 +/- 54 fmol/mg protein, respectively. Competition for /sup 125/I-salmon calcitonin binding by a wide range of calcitonin analogs revealed a close correspondence between the reported biological potencies and activities in the current system. The localization of binding sites within the nephron corresponds to the reported localization of calcitonin-stimulated adenylate cyclase activity and suggests that the receptor mediated actions of calcitonin in the kidney utilize cyclic AMP as a second messenger. In addition, the microscopic identification of specific calcitonin receptors helps the delineation of direct actions of this hormone from those which are indirect.

  3. Quantitative autoradiography of alpha particle emission in geo-materials using the Beaver™ system (United States)

    Sardini, Paul; Angileri, Axel; Descostes, Michael; Duval, Samuel; Oger, Tugdual; Patrier, Patricia; Rividi, Nicolas; Siitari-Kauppi, Marja; Toubon, Hervé; Donnard, Jérôme


    In rocks or artificial geo-materials, radioactive isotopes emitting alpha particles are dispersed according to the mineralogy. At hand specimen scale, the achievement of quantitative chemical mapping of these isotopes takes on a specific importance. Knowledge of the distribution of the uranium and thorium series radionuclides is of prime interest to several disciplines, from the geochemistry of uranium deposits, to the dispersion of uranium mill tailings in the biosphere. The disequilibrium of these disintegration chains is also commonly used for dating. However, some prime importance isotopes, such as 226Ra, are complicated to localize in geo-materials. Because of its high specific activity, 226Ra is found in very low concentrations ( ppq), preventing its accurate localization in rock forming minerals. This paper formulates a quantitative answer to the following question: at hand specimen scale, how can alpha emitters in geo-materials be mapped quantitatively? In this study, we tested a new digital autoradiographic method (called the Beaver™) based on a Micro Patterned Gaseous Detector (MPGD) in order to quantitatively map alpha emission at the centimeter scale rock section. Firstly, for two thin sections containing U-bearing minerals at secular equilibrium, we compared the experimental and theoretical alpha count rates, measured by the Beaver™ and calculated from the uranium content, respectively. We found that they are very similar. Secondly, for a set of eight homemade standards made up of a mixture of inactive sand and low-radioactivity mud, we compared the count rates obtained by the Beaver™ and by an alpha spectrometer. The results indicate (i) a linearity between both count rates, and (ii) that the count obtained by the Beaver™ can be estimated from the count obtained by the alpha spectrometry using a factor of 0.82.

  4. Autoradiography of dopamine receptors and dopamine uptake sites in the spontaneously hypertensive rat

    Energy Technology Data Exchange (ETDEWEB)

    Kujirai, K.; Przedborski, S.; Kostic, V.; Jackson-Lewis, V.; Fahn, S.; Cadet, J.L. (Columbia Univ., New York, NY (USA))


    We examined the status of dopamine (DA) D1 and D2 receptors by using (3H)SCH 23390 and (3H)spiperone binding, respectively, and DA uptake sites by using (3H)mazindol binding in spontaneously hypertensive rats (SHR) and Sprague-Dawley (SD) rats. SHR showed significantly higher (3H)SCH 23390 and (3H)spiperone binding in the caudate-putamen (CPu), the nucleus accumbens (NAc) and the olfactory tubercle (OT) in comparison to the SD rats. There were no significant differences in (3H)mazindol-labeled DA uptake sites between the two strains. Unilateral 6-hydroxydopamine (6-OHDA) injection into the striatum resulted in more than 90% depletion of DA uptake sites in the CPu in both strains. 6-OHDA-induced DA depletion was associated with significant increases in striatal (3H)spiperone binding which were of similar magnitude in the SD rats (+64.1%) and SHR (+51.3%). There were only small decreases (-5.4%) in D1 receptor binding in the dorsolateral aspect of the CPu in the SHR, whereas there were no changes in striatal D1 receptors in the SD rats. These results indicate that, although the SHR have higher concentrations of both D1 and D2 receptors in the basal ganglia, these receptors are regulated in a fashion similar to DA receptors in SD rats after 6-OHDA-induced striatal DA depletion.

  5. Co-60 and Ca-45 autoradiography in cerebral ischemia in the rat

    NARCIS (Netherlands)

    Stevens, H; Krop-van Gastel, W; Korf, J


    Radioisotopes of divalent Co (Co-57 in Single photon emission tomography (SPECT)and Co-55 in positron emission tomography (PET) have clinically been applied to visualize Ca related brain damage. The cerebral uptake of Ca-45 and Co-60 in a unilateral stroke model in the rat was compared; 100 mu Ci (C

  6. Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures

    Energy Technology Data Exchange (ETDEWEB)

    Katz, D.M.; Kimelberg, H.K.


    Primary astrocyte cultures prepared from the cerebral cortices of neonatal rats showed significant accumulation of serotonin (5-hydroxytryptamine; (/sup 3/H)-5-HT). At concentrations in the range of 0.01 to 0.7 microM (/sup 3/H)-5-HT, this uptake was 50 to 85% Na+ dependent and gave a Km of 0.40 +/- 0.11 microM (/sup 3/H)-5-HT and a Vmax of 6.42 +/- 0.85 (+/- SEM) pmol of (/sup 3/H)-5-HT/mg of protein/4 min for the Na+-dependent component. In the absence of Na+ the uptake was nonsaturable. Omission of the monoamine oxidase inhibitor pargyline markedly reduced the Na+-dependent component of (/sup 3/H)-5-HT uptake but had a negligible effect on the Na+-independent component. This suggest significant oxidative deamination of serotonin after it has been taken up by the high affinity system, followed by release of its metabolite. The authors estimated that this system enabled the cells to concentrate (/sup 3/H)-5-HT up to 44-fold at an external (/sup 3/H)-5-HT concentration of 10(-7) M. Inhibition of (/sup 3/H)-5-HT uptake by a number of clinically effective antidepressants was also consistent with a specific high affinity uptake mechanism for 5-HT, the order of effectiveness of inhibition being chlorimipramine greater than fluoxetine greater than imipramine = amitriptyline greater than desmethylimipramine greater than iprindole greater than mianserin. Uptake of (/sup 3/H)-5-HT was dependent on the presence of Cl- as well as Na+ in the medium, and the effect of omission of both ions was nonadditive. Varying the concentration of K+ in the media from 1 to 50 mM had a limited effect on (/sup 3/H)-5-HT uptake.

  7. Schistosoma mansoni: quantification of skin penetration and early migration by differential external radioassay and autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Georgi, J.R. (New York State College of Veterinary Medicine, Ithaca (USA))


    Eleven gamma-emitting radionuclides (/sup 49/Sc, /sup 54/Mn, /sup 59/Fe, /sup 60/Co, /sup 65/Zn, /sup 75/Se (as selenomethionine, selenocystine, selenite and selenate), /sup 109/Cd, /sup 125/Sb, /sup 133/Ba, /sup 137/Cs and /sup 203/Hg) were screened as labelling agents for Schistosoma mansoni cercariae by incubation of infected Biomphalaria glabrata snails in radioactive solution. Only (/sup 75/Se)methionine yielded satisfactorily labelled cercariae. Differential external radioassay, a new technique employing partial body shielding within a total body counter, permitted separate estimation of tail and body radioactivity of conscious mice previously exposed by tail immersion to /sup 75/Se-labelled cercariae, with measurements repeated as often as desired. Approximately 39% of the /sup 75/Se present in emergent cercariae was retained by schistosomula transformed in vitro but this was subject to considerable variation, especially in schistosomula transformed in vivo. Secreted or catabolized label from penetrant cercariae and schistosomula was rapidly removed from the skin by the bloodstream. Numbers of schistosomula in tail skin were directly proportional to the number of reduced silver foci counted on tail autoradiograms; only a very small fraction of tail radioactivity represented unbound ('spurious') label. Migration of schistosomula away from skin was 50% complete at 3.8-4.3 days, as determined by probit analysis of autoradiographic data.

  8. Labelling of Cells Engaged in DNA Synthesis: Autoradiography and BrdU Staining

    DEFF Research Database (Denmark)

    Madsen, Peder Søndergaard


    The cell cycle is divided in four phases: G1 phase, S phase (DNA-synthesis), G2 phase (together termed interphase) and M phase (mitosis). Cells that have ceased proliferation enter a state of quiescence called G0. M phase is itself composed of two tightly coupled processes: mitosis, in which...

  9. Molecular imaging of the serotonin 5-HT7 receptors: from autoradiography to positron emission tomography. (United States)

    Zimmer, Luc; Billard, Thierry


    Serotonin and its various receptors are involved in numerous brain functions and neuropsychiatric disorders. Of the 14 known serotoninergic receptors, the 5-HT7 receptor is the most recently identified and characterized. It is closely involved in the pathogenesis of depression, anxiety, epilepsy and pain and is therefore an important target for drug therapy. It is a crucial target in neuroscience, and there is a clear need for radioligands for in vitro and in vivo visualization and quantification, first in animal models and ultimately in humans. This review focuses on the main radioligands suggested for in vitro and in vivo imaging of the 5-HT7 receptor.

  10. Glycine uptake by lamprey spinal neurons demonstrated by light microscopic autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Sheridan, P.H.; Youngs, L.J.; Krieger, N.R.; Selzer, M.E.


    We have mapped the neuronal uptake of 3H-glycine in the spinal cords of large larval sea lampreys: Petromyzon marinus. Spinal cords were incubated in 10(-6) M 3H-glycine for 15 minutes. They were rinsed in lamprey solution, fixed in phosphate-buffered 2% glutaraldehyde, and washed in phosphate buffer. They were then sectioned with a cryostat at 16-m thickness or dehydrated, embedded in Epon, and sectioned at 1-4 micron. Sections were coated with a photographic emulsion and maintained at 4 degrees C for 1-7 days. By sectioning horizontally, it was possible to obtain complete serial reconstructions of up to 1.5-mm lengths of cord in 100-150 sections. The outlines of labelled cells were traced with a Nikon drawing attachment. For one Epon-embedded spinal cord sectioned at 4 micron, tracings were superimposed to form complete maps for 0.6-1.5-mm lengths in three representative regions of cord: rostral (gill region), caudal (dorsal fin region), and midsection. The labelled neurons were small (5-10-micron diameter) cells distributed throughout the central gray columns. They numbered 22 cells per hemisegment in the rostral region, 33 in the midsection, and 43 in the caudal region. None of the previously identified cell types were labelled, including lateral interneurons, edge cells, giant interneurons, dorsal cells, and Mueller and Mauthner axons.

  11. Quantitative determination of angiotensin II binding sites in rat brain and pituitary gland by autoradiography

    Energy Technology Data Exchange (ETDEWEB)

    Israel, A.; Correa, F.M.A.; Niwa, M.; Saavedra, J.M. (National Inst. of Mental Health, Bethesda, MD (USA))


    Rat brain and pituitary angiotensin II (AII) binding sites were quantitated by incubation of tissue sections with /sup 125/I-(Sar/sup 1/) AII, Ultrofilm radioautography, computerized densitometry, and comparison with /sup 125/I-standards at appropriate film exposure times. The highest number of AII binding sites was found in anterior pituitary and the circumventricular organs, organon subfornicalis and organon vasculosum laminae terminalis.

  12. Protracted treatment with MDMA induces heteromeric nicotinic receptor up-regulation in the rat brain: an autoradiography study. (United States)

    Ciudad-Roberts, Andrés; Camarasa, Jorge; Pubill, David; Escubedo, Elena


    Previous studies indicate that 3,4-methylenedioxy-methamphetamine (MDMA, ecstasy) can induce a heteromeric nicotinic acetylcholine receptor (nAChR, mainly of α4β2 subtype) up-regulation. In this study we treated male Sprague-Dawley rats twice-daily for 10 days with either saline or MDMA (7 mg/kg) and sacrificed them the day after to perform [(125)I]Epibatidine binding autoradiograms on serial coronal slices. MDMA induced significant increases in nAChR density in the substantia nigra, ventral tegmental area, nucleus accumbens, olfactory tubercle, anterior caudate-putamen, somatosensory, motor, auditory and retrosplenial cortex, laterodorsal thalamus nuclei, amygdala, postsubiculum and pontine nuclei. These increases ranged from 3% (retrosplenial cortex) to 30 and 34% (amygdala and substantia nigra). No increased α4 subunit immunoreactivity was found in up-regulated areas compared with saline-treated rats, suggesting a post-translational mechanism as occurs with nicotine. The heteromeric nAChR up-regulation in certain areas could account, at least in part, for the reinforcing, sensitizing and psychiatric disorders observed after long-term consumption of MDMA.

  13. Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections

    NARCIS (Netherlands)

    Knoll, G.; Burger, K.N.J.; Bron, R.; van Meer, G.; Verkleij, A.J.


    The fusion of liposomes with the plasma membrane of influenza virus-infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry, 24: 3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the apica

  14. Quantitative autoradiography of the binding sites for ( sup 125 I) iodoglyburide, a novel high-affinity ligand for ATP-sensitive potassium channels in rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Gehlert, D.R.; Gackenheimer, S.L.; Mais, D.E.; Robertson, D.W. (Eli Lilly and Co., Indianapolis, IN (USA))


    We have developed a high specific activity ligand for localization of ATP-sensitive potassium channels in the brain. When brain sections were incubated with ({sup 125}I)iodoglyburide (N-(2-((((cyclohexylamino)carbonyl)amino)sulfonyl)ethyl)-5-{sup 125}I-2- methoxybenzamide), the ligand bound to a single site with a KD of 495 pM and a maximum binding site density of 176 fmol/mg of tissue. Glyburide was the most potent inhibitor of specific ({sup 125}I)iodoglyburide binding to rat forebrain sections whereas iodoglyburide and glipizide were slightly less potent. The binding was also sensitive to ATP which completely inhibited binding at concentrations of 10 mM. Autoradiographic localization of ({sup 125}I)iodoglyburide binding indicated a broad distribution of the ATP-sensitive potassium channel in the brain. The highest levels of binding were seen in the globus pallidus and ventral pallidum followed by the septohippocampal nucleus, anterior pituitary, the CA2 and CA3 region of the hippocampus, ventral pallidum, the molecular layer of the cerebellum and substantia nigra zona reticulata. The hilus and dorsal subiculum of the hippocampus, molecular layer of the dentate gyrus, cerebral cortex, lateral olfactory tract nucleus, olfactory tubercle and the zona incerta contained relatively high levels of binding. A lower level of binding (approximately 3- to 4-fold) was found throughout the remainder of the brain. These results indicate that the ATP-sensitive potassium channel has a broad presence in the rat brain and that a few select brain regions are enriched in this subtype of neuronal potassium channels.

  15. Evaluation of the cause of unexplained radiocaesium contamination of brown rice in Fukushima in 2013 using autoradiography and gamma-ray spectrometry (United States)

    Matsunami, Hisaya; Murakami, Toshifumi; Fujiwara, Hideshi; Shinano, Takuro


    The Great East Japan Earthquake on 11 March 2011, caused the release of radioactive materials from the Fukushima Daiichi Nuclear Power Plant (FDNPP), contaminating eastern Japan, particularly in part of Fukushima Prefecture. In 2012 and 2014, the radiocaesium concentration in brown rice did not exceed regulatory levels in Minamisoma City, Fukushima. However, in 2013, some radiocaesium concentrations in brown rice exceeded regulatory levels. In this work, autoradiograms showed that high radioactivity was present as contaminated spots on the panicles of rice and in brown rice in 2013. We evaluate the contribution of direct contamination to the radiocaesium concentration in brown rice and discuss the origin of radiocaesium contamination in brown rice using the 134Cs/137Cs radioactivity ratio. Here, we show that the main cause of the unexplained radiocaesium contamination of brown rice in Minamisoma City in 2013 is the adherence of radioactive materials to the rice panicles, and these radioactive materials are associated with reactor units 2 or 3 of FDNPP.

  16. Effects of irradiation on intracellular protein synthesis in demilune of mouse sublingual gland; Study of electron microscopic autoradiography with [sup 3]H-leucine

    Energy Technology Data Exchange (ETDEWEB)

    Ohki, Ikuko; Nasu, Masanori; Matsunaga, Hajime; Furumoto, Keiichi (Nippon Dental Univ., Tokyo (Japan))


    The effects of radiation on amino acid transport in demilune of the sublingual glands were examined in mice. After 10 Gy X-ray irradiation to the submandibular region, [sup 3]H-leucine was administered to these mice. Three days after irradiation, light microscopic autoradiograms and electron microscopic autoradiograms were obtained to determine the silver grain count in demilune and the distribution of silver grain in the organelles of these cells involved in protein synthesis. Both non-irradiated and irradiated groups showed a peak count 30 minutes after the administration of [sup 3]H-leucine and a tendency to decrease in the count thereafter. The magnitude of the chronological change in this parameter was smaller in the irradiated group than in the non-irradiated group. The percentage of silver grain in the rough endoplasmic reticulum also reached a peak 30 minutes after [sup 3]H-leucine administration in both non-irradiated and irradiated groups. Thereafter the percentage tended to decrease with time in the non-irradiated group. In the irradiated group, on the other hand, the percentage showed little change from the peak level at 60, 120, and 180 minutes. In the non-irradiated group, the percentage of silver grain in the Golgi apparatus was relatively high at 30 and 60 minutes. The percentage tended to be low at 120 and 180 minutes in the irradiated group. In the non-irradiated group, the percentage of silver grain in condencing vacuoles reached a peak at 60 minutes and decreased gradually thereafter. This parameter reached a peak at 30 minutes in the irradiated group. The percentage of silver grain in the secretory granules increased with time in both non-irradiated and irradiated groups. At 30 and 60 minutes, the percentage was significantly higher in the irradiated group than in the non-irradiated group. At 180 minutes, however, it was significantly lower in the irradiated group than in the non-irradiated group. (author).

  17. [O-methyl-{sup 11}C]{beta}-CIT-FP, a potential radioligand for quantitation of the dopamine transporter: Preparation, autoradiography, metabolite studies, and positron emission tomography examinations

    Energy Technology Data Exchange (ETDEWEB)

    Lundkvist, Camilla; Halldin, Christer; Swahn, Carl-Gunnar; Hall, Haakan; Karlsson, Per; Nakashima, Yoshifumi; Wang, Shaoyin; Milius, Richard A.; Neumeyer, John L.; Farde, Lars


    {beta}-CIT-FP [N-(3-fluoropropyl)-2{beta}-carbomethoxy-3{beta}-(4-iodophenyl)nortropane] is a cocaine analogue with a high affinity for the dopamine transporter. [O-methyl-{sup 11}C]{beta}-CIT-FP ([{sup 11}C]{beta}-CIT-FP) was prepared byO -alkylation of the free acid with [{sup 11}C]methyl iodide. The total radiochemical yield of [{sup 11}C]{beta}-CIT-FP was 50 to 60% with an overall synthesis time of 30 min. The radiochemical purity was >99%, and the specific radioactivity at time of injection was about 37 GBq/{mu}mol (1000 Ci/mmol). Autoradiographic examination of [{sup 11}C]{beta}-CIT-FP binding in human brain postmortem demonstrated specific binding in the caudate nucleus and putamen. Positron emission tomography (PET) examination of [{sup 11}C]{beta}-CIT-FP in a Cynomolgus monkey demonstrated accumulation in the striatum with a striatum-to-cerebellum ratio of about 8 after 60 min. Equilibrium in the striatum was attained within 70 to 90 min. The radioactivity ratios of thalamus/cerebellum and neocortex/cerebellum were about 2 and 1.5, respectively. In a displacement experiment, radioactivity in the striatum but not in the cerebellum was reduced after injection of {beta}-CIT, indicating that striatal radioactivity following injection of [{sup 11}C]{beta}-CIT-FP is associated with dopamine transporter sites and that the binding is reversible. The fraction of the total radioactivity in plasma representing [{sup 11}C]{beta}-CIT-FP determined by high-performance liquid chromatography (HPLC) was 84% at 15 min and 50% at 95 min. [{sup 11}C]{beta}-CIT-FP should be a useful PET radioligand for the quantitation of dopamine transporters in the human brain in vivo.

  18. [Study of regeneration in periodontal tissue after implantation of bone ceramic and collagen gel compound materials. Evaluation of histopathological finding and autoradiography]. (United States)

    Miyamoto, Y; Hayashi, H; Kamoi, K


    The aim of this study is to determine the process of periodontal tissue regeneration and the metabolic activity of osteoblasts after implantation of bone ceramic and collagen gel compound materials (BC). Bone defects were artificially prepared in the alveolar septa of the bilateral upper first and second molars of Wistar rats. Subsequently, BC were implanted into the defective sites on the left side, and the gingival flaps were closed. At the defective sites on the right side, as a control, gingival flaps were closed without implantation. Rats were sacrificed 1, 3, 5, 7 or 14 weeks after implantation, and prepared tissue sections were observed both pathologically and autoradiographically using 3H-Proline. The results obtained were as follows: Pathological Findings One week after BC implantation, inflammatory cellular infiltration of the surrounding gingival connective tissue was relatively mild. Three weeks after implantation, BC were present in fibrous connective tissues, and some directly bound to the marices of regenerated bone. Observation 5 weeks after implantation revealed that BC had become embedded in the regenerated bone matrices and that there was giant cell reaction to foreign bodies at the margin of BC located in connective tissue. BC were directly bound to the regenerated bone matrices without intermediary fibrous tissues 7 and 14 weeks after implantation. Connective tissues showed high grade regeneration of collagen fiber bundles, in an arrangement that tended to be fixed in mesial and distal directions. Autoradiographic Findings There was no uptake of 3H-Proline into the regenerated bone matrices or the gingival connective tissue surrounding BC, while uptake of 3H-Proline into the entire area around the root apex and in the vicinity of the alveolar septum was observed with time (weeks) after BC implantation. These results suggest that BC provide nuclei for bone regeneration through inclusion in newly-generated periodontal bone tissue, although it is difficult to produce definite induction of bone tissue by BC alone. It is also apparent that these are useful bone implantation materials for restoration of the physiological morphology of alveolar bone in periodontal surgical treatment.

  19. Imaging Hypoxia in Xenografted and Murine Tumors with 18F-Fluoroazomycin Arabinoside: A Comparative Study Involving microPET, Autoradiography, pO2-Polarography, and Fluorescence Microscopy

    DEFF Research Database (Denmark)

    Busk, Morten; Horsman, Michael R; Jakobsen, Steen;


    PURPOSE: Positron emission tomography (PET) allows noninvasive assessment of tumor hypoxia; however the combination of low resolution and slow tracer clearance from nonhypoxic tissue is problematic. The aim of this study was to examine the in vivo hypoxia selectivity of fluoroazomycin arabinoside...... ([(18)F]-FAZA), a promising tracer with improved washout kinetics from oxygenated tissue. METHODS AND MATERIALS: Three squamous cell carcinomas and one fibrosarcoma with widely differing spatial patterns of vascularization, hypoxia, and necrosis were grown in mice and evaluated with PET...

  20. Regional distribution of putative NPY Y*U1 receptors and neurons expressing Y*U1 mRNA in forebrain areas of the rat central nervous system

    DEFF Research Database (Denmark)

    Larsen, Philip J.; Sheikh, Søren P.; Jakobsen, Cherine R.


    Anatomi, neurobiologi, neuropeptide Y, NPY analogues, receptor autoradiography, in situ hybridization histochemistry, Y*U1 mRNA, Y*U1 andY*U2 receptors, rat......Anatomi, neurobiologi, neuropeptide Y, NPY analogues, receptor autoradiography, in situ hybridization histochemistry, Y*U1 mRNA, Y*U1 andY*U2 receptors, rat...

  1. Expression of receptors for gut peptides in pancreata of BOP-treated and control hamsters

    NARCIS (Netherlands)

    Tang, C.; Biemond, I.; Appel, M.J.; Visser, C.J.T.; Woutersen, R.A.; Lamers, C.B.H.W.


    The growth of pancreatic cancers may be influenced by certain gut peptides. However, the alteration of gut peptide receptors in the progress of pancreatic carcinogenesis is largely unknown. With storage phosphor autoradiography, this study visualized and characterized receptors for cholecystokinin (


    Institute of Scientific and Technical Information of China (English)

    CHUJin-Shong; WANGYi-Fei


    The specific estrogen binding site for 17β-estradiol has been investigated on human spermatozoa by electron microscopec autoradiography. The results show that the binding sites were distributed over the surface of human spermatozoa: acrosomal cap, equatorial

  3. DNA and RNA Synthesis in Animal Cells in Culture--Methods for Use in Schools (United States)

    Godsell, P. M.; Balls, M.


    Describes the experimental procedures used for detecting DNA and RNA synthesis in xenopus cells by autoradiography. The method described is suitable for senior high school laboratory classes or biology projects, if supervised by a teacher qualified to handle radioisotopes. (JR)

  4. Quantitative analysis of autoradiographic image intensification using Thiourea-S35 (United States)

    Askins, B. S.; Odell, C. R.


    Photographic images enhanced by the method of Thiourea-S35 autoradiography are evaluated in terms of signal-to-noise ratio, detective quantum efficiency (DQE), and Wiener spectrum analysis using digitized images. It is determined that the original signal-to-noise ratio is not degraded by the intensification process which allows an increase in the practical working DQE as a function of density. These results apply at all spatial frequencies that were tested. The advantage given by autoradiography is the ability to produce usable images from emulsions originally exposed to the low densities corresponding to maximum DQE and movement of faint image densities above the level of the threshold for detection.

  5. Modulation of Molecular Markers by CLA. (United States)


    electrophoresis on a 6% polyacrylamide gel and detected by autoradiography using Kodak X- ray films. Results and Discussion Appendix 1 contains four...floc 6.5 Corn starch 18.2 Cerelose 18.2 Casein 26.0 Vitamin mixb (vitamin E omitted) 1.30 Mineral mix’ 4.55 DL-a-tocopherol acetate (LPX diets only...used to label the PCR products. The digested products were separated by electrophoresis on a 6% polyacrylamide gel and detected by autoradiography

  6. Catechol-O-methyltransferase: a method for autoradiographic visualization of isozymes in cellogel

    Energy Technology Data Exchange (ETDEWEB)

    Brahe, C.; Crosti, N.; Meera Khan, P.; Serra, A.


    An electrophoretic procedure for separating the molecular forms of catechol-O-methyltransferase in cellulose acetate gel is described; the zones of enzyme activity were revealed by autoradiography. The electrophoretic patterns of the enzyme in several tissues and cell lines derived from four different species are presented.

  7. Zolmitriptan (a 5-HT1B/1D receptor agonist with central action) does not increase symptoms in obsessive compulsive disorder

    NARCIS (Netherlands)

    Boshuisen, ML; den Boer, JA


    Rationale: Non-selective serotonin (5-HT) receptor agonists like meta-chlorophenylpiperazine and MK-212 have been used to explore the role of 5-HT in obsessive compulsive disorder (OCD). The results of these studies and the findings of autoradiography and neuroimaging studies, pointed to a possible

  8. {sup 11}C-harmine as a potential PET tracer for ductal pancreas cancer: in vitro studies

    Energy Technology Data Exchange (ETDEWEB)

    Herlin, G.; Persson, B.; Laangstroem, B.; Aspelin, P. [Department of Diagnostic Radiology, Huddinge University Hospital, 141-86 Stockholm (Sweden); Bergstroem, M. [Uppsala University PET Centre, Uppsala (Sweden)


    Our objective was to find a tracer in diagnosing human pancreatic cancer using positron emission tomography (PET). For this purpose in vitro test of pancreatic tissues with autoradiography was used. Autoradiography was performed with {sup 11}C-harmine (a MAO-A-inhibitor) with and without competitive inhibition. Tissue preparations were obtained from normal human pancreas and pancreatic cancer. The uptake was compared with rat brain or pig brain, tissues with high expression of MAO-A. Nine autoradiography studies on 16 samples from five different human pancreatic cancers gave a significant level of specific binding of {sup 11}C-harmine in 13, and 3 samples did not give a significant level of specific binding of {sup 11}C-harmine. All 16 samples were analysed with autoradiography. Compared with rat brain, the uptake in the human cancers varied between 9 and 43% except for one tissue preparation which had a too low value for measurement. This study shows expression of MAO-A in human pancreatic cancer. This is readily characterised in vitro. The potential use of {sup 11}C-harmine in the diagnosis of pancreatic cancer using PET might be limited, but further PET studies are necessary. (orig.)

  9. 11C-harmine as a potential PET tracer for ductal pancreas cancer: in vitro studies. (United States)

    Herlin, G; Persson, B; Bergström, M; Långström, B; Aspelin, P


    Our objective was to find a tracer in diagnosing human pancreatic cancer using positron emission tomography (PET). For this purpose in vitro test of pancreatic tissues with autoradiography was used. Autoradiography was performed with (11)C-harmine (a MAO-A-inhibitor) with and without competitive inhibition. Tissue preparations were obtained from normal human pancreas and pancreatic cancer. The uptake was compared with rat brain or pig brain, tissues with high expression of MAO-A. Nine autoradiography studies on 16 samples from five different human pancreatic cancers gave a significant level of specific binding of (11)C-harmine in 13, and 3 samples did not give a significant level of specific binding of (11)C-harmine. All 16 samples were analysed with autoradiography. Compared with rat brain, the uptake in the human cancers varied between 9 and 43% except for one tissue preparation which had a too low value for measurement. This study shows expression of MAO-A in human pancreatic cancer. This is readily characterised in vitro. The potential use of (11)C-harmine in the diagnosis of pancreatic cancer using PET might be limited, but further PET studies are necessary.

  10. 75 FR 24428 - Spirodiclofen; Pesticide Tolerances (United States)


    ..., Leydig cell adenoma in male rats, and liver tumors in mice. EPA classified spirodiclofen as ``likely to... male rats, uterine adenomas and/or adenocarcinoma in female rats, and liver tumors in mice... urine and feces. A rat whole body autoradiography study showed no accumulation in any specific organs...


    DEFF Research Database (Denmark)

    Glud, Andreas Nørgaard; Landau, A.M.; Johnsen, Erik Lisbjerg


    and histology. Evaluation of gait, PET, autoradiography and histology are ongoing on AAV-models and awaiting on inoculation fibril-models. Discussion: We predict that these animal models will be useful and beneficial in the understanding of pathological mechanisms of human PD, novel therapeutic strategies...

  12. Neuroexcitatory Drug Receptors in Mammals and Invertebrates (United States)


    T.A. Miller and R.W. Olsen (1988) Quantitative autoradiography of GABA receptors in locust (Schistocerca americana). Brain Pestic . Sci. 24, 299-309. 6... Pestic . Scl. 24, 299-309 (1988). Olsen, R.W., Szamraj, 0. and Miller, T. [35S]t-Butyl Bicyclophosphorothionate (TBPS) Binding Sites in Invertebrate

  13. Research Training Program in Breast Cancer (United States)


    release from CF~s in a form that then activates histone gene transcription. Many Nucleosoines Histones transcription factors accumulate in inactive pools ...nitrocellulose before immunob- Cajal, S.R.Y. 1903. Un sencillo metodo de coloracion seletiva lotting and autoradiography. del reticulo

  14. Effect of acute whole-body neutron gamma irradiation on the dopamine neuronal uptake-sites; Effets d`une irradiation globale aigue a preponderance neutron sur le transporteur de capture neuronale de la dopamine

    Energy Technology Data Exchange (ETDEWEB)

    Martin, C.; Mahfoudi, H.; Lambert, F.; Burckhart, M.F.; Fatome, M. [Centre de Recherches du Service de Sante des Armees, La Tronche, 38 - Grenoble (France)


    The effects of (neutron-gamma) irradiation on the dopamine uptake sites distribution were investigated, using quantitative autoradiography. Brain ares examined are striatum, lateral septum, substantia nigra, gyrus dentatus, ventral tegmental area, interfascicular nu and antero-ventral thalamic nu. Three hours after exposure at the dose of 4 Gy, a decrease (- 33 %) of dopamine uptake sites was observed in the gyrus dentatus. (authors)

  15. Changes in 5-HT1A and NMDA binding sites by a single rapid transcranial magnetic stimulation procedure in rats

    NARCIS (Netherlands)

    Kole, MHP; Fuchs, E; Ziemann, U; Paulus, W; Ebert, U


    The effects of a single rapid-rate transcranial magnetic stimulation (rTMS) exposure on neurotransmitter binding sites in the rat brain 24 h after the stimulation were examined. Quantification by in vitro-autoradiography showed no differences for H-3-paroxetine binding (5-HT uptake sites) between rT

  16. Kinetics of small lymphocytes in normal and nude mice after splenectomy

    DEFF Research Database (Denmark)

    Hougen, H P; Hansen, F; Jensen, E K


    Autoradiography and various quantitations on lymphoid tissues have been used to evaluate the kinetics of small lymphocytes in normal (+/nu or +/+) and congenitally athymic nude (nu/nu) NMRI mice 1 month after splenectomy or sham-splenectomy. The results indicate that splenectomy causes depressed...

  17. Radiosynthesis and characterisation of a potent and selective GPR139 agonist radioligand

    DEFF Research Database (Denmark)

    Kuhne, Sebastiaan; Nøhr, Anne Cathrine; Marek, AleŠ;


    Compound 1 is a selective and potent agonist of the G protein-coupled receptor GPR139 (EC50 = 39 nM). In this study, we describe the synthesis, radiolabelling and in vitro evaluation of [3H]-1 for the characterisation of GPR139 and its spatial expression in the brain using autoradiography. Two di...

  18. Rembrandt’s An Old Man in Military Costume: the underlying image re-examined

    NARCIS (Netherlands)

    Trentelman, K.; Janssens, K.; Van der Snickt, G.; Szafran, Y.; Woollett, A.T.; Dik, J.


    The painting An Old Man in Military Costume in the J. Paul Getty Museum, by Rembrandt Harmensz van Rijn, was studied using two complementary, element-specific imaging techniques—neutron activation autoradiography (NAAR) and macro-X-ray fluorescence (MA-XRF) mapping—to reveal the second, hidden paint

  19. Autoradiographic studies in a rabbit osteoarthrosis model

    Energy Technology Data Exchange (ETDEWEB)

    Fengler, H.; Franz, R. (Medizinische Akademie, Dresden (German Democratic Republic))


    To study the onset of the osteoarthrotic process, an osteoarthrosis model was used on the knee joint in adult rabbits by a valgus deformity of the proximal tibia of 30/sup 0/ in conformity with Reimann 1973. The synthesis capacity of the chondrocytes was investigated by using /sup 35/S-sulfate autoradiographies. Already prior to the affection of the superficial integrity of the cartilage it was possible to observe an enhanced glycosaminoglycan synthesis, but with progressing fibrillation the sulfate incorporation was found to be diminished. Thanks to autoradiography with /sup 3/H-thymidine the replication of the chondrocytes was already found at very early stages of osteoarthrosis that is likely to occur mitotically. The osteotomy itself induces mitoses and an enhanced glycosaminoglycan synthesis.

  20. Autoradiographic localization of estrogen binding sites in human mammary lesions

    Energy Technology Data Exchange (ETDEWEB)

    Buell, R.H.


    The biochemical assay of human mammary carcinomas for estrogen receptors is of proven clinical utility, but the cellular localization of estrogen binding sites within these lesions is less certain. The author describes the identification of estrogen binding sites as visualized by thaw-mount autoradiography after in vitro incubation in a series of 17 benign and 40 malignant human female mammary lesions. The results on the in vitro incubation method compared favorably with data from in vivo studies in mouse uterus, a well-characterized estrogen target organ. In noncancerous breast biopsies, a variable proportion of epithelial cells contained specific estrogen binding sites. Histologically identifiable myoepithelial and stromal cells were, in general, unlabeled. In human mammary carcinomas, biochemically estrogen receptor-positive, labeled and unlabeled neoplastic epithelial cells were identified by autoradiography. Quantitative results from the autoradiographic method compared favorably with biochemical data.

  1. A new, fast and semi-automated size determination method (SASDM for studying multicellular tumor spheroids

    Directory of Open Access Journals (Sweden)

    Lindhe Örjan


    Full Text Available Abstract Background Considering the width and importance of using Multicellular Tumor Spheroids (MTS in oncology research, size determination of MTSs by an accurate and fast method is essential. In the present study an effective, fast and semi-automated method, SASDM, was developed to determinate the size of MTSs. The method was applied and tested in MTSs of three different cell-lines. Frozen section autoradiography and Hemotoxylin Eosin (H&E staining was used for further confirmation. Results SASDM was shown to be effective, user-friendly, and time efficient, and to be more precise than the traditional methods and it was applicable for MTSs of different cell-lines. Furthermore, the results of image analysis showed high correspondence to the results of autoradiography and staining. Conclusion The combination of assessment of metabolic condition and image analysis in MTSs provides a good model to evaluate the effect of various anti-cancer treatments.

  2. Characterization of hydrofracture grouts for radionuclide migration

    Energy Technology Data Exchange (ETDEWEB)

    Stinton, D.P.; McDaniel, E.W.; Weeren, H.O.


    Detailed characterization of hydrofracture grouts was performed by optical microscopy, scanning electron microscopy, x-ray diffraction, and ..beta..-..gamma.. autoradiography. Laboratory-produced samples containing simulated wastes as well as actual radioactive samples of hydrofracture grout sheets obtained by core drilling were examined in this work. X-ray diffraction results revealed that both laboratory-produced samples and a core-drilled sample consisted primarily of calcium carbonate phases. Both sample types contained very small amounts of strontium or cesium wastes, neither of which could be detected by microscopic techniques. The core-drilled sample contained radioactive /sup 90/Sr, /sup 137/Cs, and /sup 60/Co that could be detected by ..beta..-..gamma.. autoradiography. The autoradiograph revealed that these radionuclides were still present in the 20-year-old grout and that they had not migrated into the trapped shale fragments.

  3. MALDI imaging mass spectrometry: bridging biology and chemistry in drug development. (United States)

    Castellino, Stephen; Groseclose, M Reid; Wagner, David


    Our understanding of drug tissue distribution impacts a number of areas in drug development, including: pharmacology, pharmacokinetics, safety, drug-drug interactions, transport and metabolism. Despite their extensive use, autoradiography and tissue homogenate LC-MS analysis have limitations in providing a comprehensive assessment of tissue distributions. In the case of autoradiography, it is the inability to distinguish between parent drug and drug metabolites. In LC-MS analysis of tissue homogenate, all tissue localization information is lost. The emerging technique of MALDI imaging mass spectrometry has the capability to distinguish between parent and metabolites while maintaining spatial distribution in tissues. In this article, we will review the MALDI imaging MS methodology as applied to drug development and provide examples highlighting the impact of this important technique in drug development.

  4. Analysis of Cytosolic Proteins that Bind to the 5' Leader Sequence of the Angiotensin AT1 Receptor by RNA Electromobility Shift Assay. (United States)

    Wu, Z; Krishnamurthi, K; Mok, K; Sandberg, K


    Electromobility shift assays (EMSAs) provide a way to study proteinnucleic acid interactions. This method is based on the observation that the electrophoretic mobility of nucleic acids through polyacrylamide gels is retarded when bound to proteins. The mobility of nucleic acid-protein complexes are thus "shifted" with respect to the free nucleic acids. Typically, the nucleic acids are labeled with (32)P. Once the nucleic acid-protein complexes are separated from free radiolabeled nucleic acids, the electrophoresis is terminated and the gel dried. The radiolabeled nucleic acids in their free and complexed forms are visualized and quantified by phosphor autoradiography or by X-ray autoradiography. DNA-binding proteins are commonly identified by EMSA. EMSA also works well for studying purified RNA-binding proteins (1-3) and this technique is currently being developed for identifying unknown RNA-binding proteins.

  5. A method for the assessment of specific energy distribution in a model tumor system

    Energy Technology Data Exchange (ETDEWEB)

    Noska, M.A.


    Due to the short range of alpha particles in tissue, the calculation of dose from internally deposited alpha emitters requires a detailed analysis of the microscopic distribution of the radionuclide in order to determine the spatial distribution of energy emission events and, from this, the spatial distribution of dose. In the present study, the authors used quantitative autoradiography (QAR) to assess the microdistribution of a radiolabeled monoclonal antibody (MAb) fragment in human glioma xenografts in mice.

  6. Chronic Opioid Therapy and Opioid Tolerance: A New Hypothesis


    Goldberg, Joel S.


    Opioids are efficacious and cost-effective analgesics, but tolerance limits their effectiveness. This paper does not present any new clinical or experimental data but demonstrates that there exist ascending sensory pathways that contain few opioid receptors. These pathways are located by brain PET scans and spinal cord autoradiography. These nonopioid ascending pathways include portions of the ventral spinal thalamic tract originating in Rexed layers VI–VIII, thalamocortical fibers that proje...

  7. Method for the typing of Clostridium difficile based on polyacrylamide gel electrophoresis of (/sup 35/S)methionine-labeled proteins

    Energy Technology Data Exchange (ETDEWEB)

    Tabaqchali, S.; O' Farrell, S.; Holland, D.; Silman, R.


    A typing method for Clostridium difficile based on the incorporation of (/sup 35/S)methionine into cellular proteins, their separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their visualization by autoradiography is described. On analysis of the radiolabeled-protein profiles, nine distinct groups were observed (A to E and W to Z). The method, which is simple, reproducible, and readily expandable, has been applied in epidemiological studies to demonstrate cross-infection and hospital acquisition of C. difficile.

  8. Spatiotemporal variability in archaeal communities of tropical coastal waters

    Digital Repository Service at National Institute of Oceanography (India)

    Singh, S.K.

    (Woese 1987), have changed our perceptions on their bio- diversity, distribution and function in natural marine ecosys- tems. Meticulous and extensive analyses of ribosomal RNA gene sequences from environmental samples have revealed that archaea.... (2001) concluded that there are 1.3×1028 archaeal cells (of which ∼20 % are thaumarchaeotes) and 3.1×1028 bacterial cells in the world oceans. A combination of in-situ hybrid- ization and micro-autoradiography has shown that marine archaea are active...

  9. Localisation and mechanism of renal retention of radiolabelled somatostatin analogues

    Energy Technology Data Exchange (ETDEWEB)

    Melis, Marleen; Krenning, Eric P.; Bernard, Bert F.; Jong, Marion de [Erasmus MC, Department of Nuclear Medicine, Rotterdam (Netherlands); Barone, Raffaella [UCL, Centre of Nuclear Medicine and Laboratory of PET, Brussels (Belgium); Visser, Theo J. [Erasmus MC, Department of Internal Medicine, Rotterdam (Netherlands)


    Radiolabelled somatostatin analogues, such as octreotide and octreotate, are used for tumour scintigraphy and radionuclide therapy. The kidney is the most important critical organ during such therapy owing to the reabsorption and retention of radiolabelled peptides. The aim of this study was to investigate in a rat model both the localisation and the mechanism of renal uptake after intravenous injection of radiolabelled somatostatin analogues. The multi-ligand megalin/cubilin receptor complex, responsible for reabsorption of many peptides and proteins in the kidney, is an interesting candidate for renal endocytosis of these peptide analogues. For localisation studies, ex vivo autoradiography and micro-autoradiography of rat kidneys were performed 1-24 h after injection of radiolabelled somatostatin analogues and compared with the renal anti-megalin immunohistochemical staining pattern. To confirm a role of megalin in the mechanism of renal retention of [{sup 111}In-DTPA]octreotide, the effects of three inhibitory substances were explored in rats. Renal ex vivo autoradiography showed high cortical radioactivity and lower radioactivity in the outer medulla. The distribution of cortical radioactivity was inhomogeneous. Micro-autoradiography indicated that radioactivity was only retained in the proximal tubules. The anti-megalin immunohistochemical staining pattern showed a strong similarity with the renal [{sup 111}In-DTPA]octreotide ex vivo autoradiograms. Biodistribution studies showed that co-injection of positively charged d-lysine reduced renal uptake to 60% of control. Sodium maleate reduced renal [{sup 111}In-DTPA]octreotide uptake to 15% of control. Finally, cisplatin pre-treatment of rats reduced kidney uptake to 70% of control. Renal retention of [{sup 111}In-DTPA]octreotide is confined to proximal tubules in the rat kidney, in which megalin-mediated endocytosis may play an important part. (orig.)

  10. Differential Regulation of α7 Nicotinic Receptor Gene (CHRNA7) Expression in Schizophrenic Smokers


    Mexal, Sharon; Berger, Ralph; Logel, Judy; Ross, Randal G.; Freedman, Robert; Leonard, Sherry


    The α7 neuronal nicotinic receptor gene (CHRNA7) has been implicated in the pathophysiology of schizophrenia by genetic and pharmacological studies. Expression of the α7* receptor, as measured by [125I]α-bungarotoxin autoradiography, is decreased in postmortem brain of schizophrenic subjects compared to non-mentally ill controls. Most schizophrenic patients are heavy smokers, with high levels of serum cotinine. Smoking changes the expression of multiple genes and differentially regulates gene...

  11. Cellular localization of AT1 receptor mRNA and protein in normal placenta and its reduced expression in intrauterine growth restriction. Angiotensin II stimulates the release of vasorelaxants.


    Li, X.; M Shams; Zhu, J; Khalig, A; Wilkes, M; Whittle, M; Barnes, N; Ahmed, A.


    Angiotensin II (ANG II) is a potent vasoconstrictor and growth promoter. Quantitative receptor autoradiography using the nonselective radioligand [125I]ANG II and subtype-selective competing compounds demonstrated the presence of both ANG II receptor (AT)1 and AT2 receptor recognition sites. In addition, a relatively small population of apparently non-AT1/non-AT2 sites was identified that may represent a novel high affinity ANG II recognition site in human placenta. Using placental membrane p...

  12. Excitatory amino acid binding sites in the hippocampal region of Alzheimer's disease and other dementias.



    Quantitative receptor autoradiography was used to measure muscarinic cholinergic, benzodiazepine, kainate, phencyclidine (PCP), N-methyl-D-aspartate (NMDA) (measured in Tris acetate), quisqualate-sensitive, non-quisqualate-sensitive and total glutamate (measured in Tris chloride buffer) binding sites in adjacent sections of the hippocampal region of 10 Alzheimer's disease, nine control, and six demented, non-Alzheimer's disease postmortem human brains. The measurements were compared to the nu...




    Portions of an adult human lung were studied by autoradiography in order to detect the presence of fallout particles. The radioactivity in the remainder of the tissue was determined with a gamma-ray spectrometer. Four particles were found and their activities were determined. From the measurement for total-fission-product activity in the lung tissue it was calculated that there were approximately 264 particles in the right lung at the time of death.

  14. Epiminocyclohepta[b]indole analogs as 5-HT6 antagonists

    DEFF Research Database (Denmark)

    Henderson, Alan J; Guzzo, Peter R; Ghosh, Animesh;


    A new series of epiminocyclohepta[b]indoles with potent 5-HT(6) antagonist activity were discovered and optimized using in vitro protocols. One compound from this series was progressed to advanced pharmacokinetic (PK) studies followed by 5-HT(6) receptor occupancy studies. The compound was found...... to have excellent oral absorption, a highly favorable PK profile and demonstrated pharmacodynamic interaction with the 5-HT(6) receptor as shown by ex vivo autoradiography....

  15. Digital imaging of autoradiographs from paintings by Georges de La Tour (1593-1652)

    CERN Document Server

    Fischer, C O; Laurenze, C; Schmidt, C; Slusallek, K


    The artistic work of the painter Georges de La Tour has been studied very intensively in the last few years, mainly by French and US-American art historians and natural scientists. To support the in-depth analysis of two paintings from the Kimbell Art Museum in Fort Worth, Texas, USA, two similar paintings from the Gemaeldegalerie Berlin have been investigated. The method of neutron activation autoradiography has been applied using imaging plates with digital image processing.

  16. Specific binding of photoaffinity-labeling peptidomimetics of Pro-Leu-Gly-NH2 to the dopamine D2L receptor: evidence for the allosteric modulation of the dopamine receptor. (United States)

    Mann, Amandeep; Verma, Vaneeta; Basu, Dipannita; Skoblenick, Kevin J; Beyaert, Michael G R; Fisher, Abigail; Thomas, Nancy; Johnson, Rodney L; Mishra, Ram K


    The present study was undertaken to investigate the mechanistic role of l-prolyl-l-leucyl-glycinamide (PLG) in modulating agonist binding to the dopamine D(2L) receptor. Competition and displacement assays indicate that the photoaffinity-labeling peptidomimetics of PLG, 3(R)-[(4(S)-(4-azido-2-hydroxy-benzoyl) amino-2(S)-pyrrolidinylcarbonyl)amino]-2-oxo-1-pyrrolidineacetamide hydrochloride (1a) and 3(R)-[(4(S)-(4-azido-2-hydroxy-5-iodo-benzoyl)amino-2(S)-pyrrolidinylcarbonyl)amino]-2-oxo-1-pyrrolidineacetamide hydrochloride (1b) bind at the same site as PLG. Autoradiography was used to establish the covalent binding of [(125)I]-1b to an approximately 51kDa protein in bovine striatal membranes. Western blot analysis with a dopamine D(2L)-specific antibody, in combination with autoradiography, following a two-dimensional gel separation, suggested this approximately 51kDa protein to be the dopamine D(2L) receptor. Further evidence for binding of 1b to dopamine D(2L) was provided by samples immunoprecipitated with the D(2L) antibody. These samples were analyzed by western blotting in parallel with autoradiography of [(125)I]-1b labeled protein. Both methods revealed bands at approximately 51kDa. Furthermore, PLG is shown to compete with 1b for binding to the dopamine D(2L) receptor as determined by autoradiography, as well as competition experiments with PLG and 1a. Collectively, these findings suggest the successful development of a photoaffinity-labeling agent, compound 1b, that has been used to elucidate the interaction of PLG specifically with the dopamine D(2L) receptor.

  17. In vitro uptake of /sup 14/C-praziquantel by cestodes, trematodes, and a nematode

    Energy Technology Data Exchange (ETDEWEB)

    Andrews, P.; Thomas, H.; Weber, H.


    /sup 14/C-praziquantel was rapidly taken up by Schistosoma mansoni, Fasciola hepatica, Hymenolepis nana, and isolated strobilocerci of Taenia taeniaeformis. Schistosoma mansoni lost praziquantel rapidly to drug-free medium. Chromatography of extracts prepared after incubation of S. mansoni and H. nana yielded no indication that praziquantel was metabolized. Autoradiography revealed a uniform distribution of praziquantel throughout the tissues of S. mansoni and H. nana. Uptake was considerably slower in the nematode Heterakis spumosa and apparently via the oral route.

  18. Regional cerebral glucose utilization during morphine withdrawal in the rat.


    Wooten, G.F.; DiStefano, P.; Collins, R. C.


    Regional cerebral glucose utilization was studied by 2-deoxy[14C]glucose autoradiography in morphine-dependent rats and during naloxone-induced morphine withdrawal. In morphine-dependent rats, glucose utilization was increased compared with naive controls uniformly (23-54%) in hippocampus, dentate gyrus, and subiculum and reduced in frontal cortex, striatum, anterior ventral thalamus, and medial habenular nucleus. On precipitation of morphine withdrawal by subcutaneous administration of nalox...

  19. Kinetics of small lymphocytes in normal and nude mice after splenectomy

    DEFF Research Database (Denmark)

    Hougen, H P; Hansen, F; Jensen, E K;


    Autoradiography and various quantitations on lymphoid tissues have been used to evaluate the kinetics of small lymphocytes in normal (+/nu or +/+) and congenitally athymic nude (nu/nu) NMRI mice 1 month after splenectomy or sham-splenectomy. The results indicate that splenectomy causes depressed ......, are released from the lympho-myeloid organs in compensation for the loss of long-lived, thymus-derived cells....

  20. 5-HT2A and mGlu2 receptor binding levels are related to differences in impulsive behavior in the Roman Low- (RLA) and High- (RHA) avoidance rat strains

    DEFF Research Database (Denmark)

    Klein, A B; Ultved, L; Adamsen, D


    of serotonin transporter (SERT), 5-HT(2A) and 5-HT(1A) receptor binding using highly specific radioligands ((3)H-escitalopram, (3)H-MDL100907 and (3)H-WAY100635) and mGlu2/3 receptor binding ((3)H-LY341495) using receptor autoradiography in fronto-cortical sections from RLA-I (n=8) and RHA-I (n=8) male rats...

  1. Examination of the translocation of sulfonylurea herbicides in sunflower plants by matrix-assisted laser desorption/ionisation mass spectrometry imaging. (United States)

    Anderson, David M G; Carolan, Vikki A; Crosland, Susan; Sharples, Kate R; Clench, Malcolm R


    Pesticides are widely used in agriculture to control weeds, pests and diseases. Successful control is dependent on the compound reaching the target site within the organism after spray or soil application. Conventional methods for determining uptake and movement of herbicides and pesticides include autoradiography, liquid scintillation and chromatographic techniques such as high-performance liquid chromatography (HPLC). Autoradiography using radiolabelled compounds provides the best indication of a compound's movement within the plant system. Autoradiography is an established technique but it relies on the synthesis of radiolabelled compounds. The distribution of four sulfonylurea herbicides in sunflower plants has been studied 24  h after foliar application. The use of matrix-assisted laser desorption/ionisation mass spectrometry imaging (MALDI-MSI) images of protonated molecules and fragment ions (resulting from fragmentation at the urea bond within the sulfonylurea herbicides) has provided evidence for translocation above and below the application point. The translocation of nicosulfuron and azoxystrobin within the same plant system has also been demonstrated following their application to the plant stem. This study provides evidence that MALDI-MSI has great potential as an analytical technique to detect and assess the foliar, root and stem uptake of agrochemicals, and to reveal their distribution through the plant once absorbed and translocated.

  2. Use of /sup 75/Se tracer and autoradiographic techniques in the study of schistosomiasis

    Energy Technology Data Exchange (ETDEWEB)

    Chandiwana, S.K. (New York State Veterinary Coll., Ithaca, NY (USA))


    The paper provides an overview of recent studies on the use of /sup 75/Se to tag larval schistosomes and to monitor their migration and distribution patterns in naive mice and those previously exposed to cercariae. The principles and techniques of radioassay and autoradiography in studying various aspects of /sup 75/Se-labelled larval schistosomes are described. The main shortcoming of radioassay in monitoring location and movement of labelled schistosomula is that some of the label dissociates from the schistosomula and accumulates in host tissues, notably the liver. Dissociated label is indistinguishable from schistosomula-bound label making monitoring of parasite migration extremely difficult. This difficulty is overcome by compressed tissue autoradiography where labelled schistosomula can be seen as reduced silver foci on an autoradiographic film, whereas dissociated label is too diffusely distributed to produce such reduced silver foci. Furthermore, using autoradiography, quantitative information on parasite migration in normal and immunized laboratory animals can be obtained that would be impossible using traditional recovery techniques. In addition to using /sup 75/Se tracer in migration studies, the radio-isotope has potential for elucidating various aspects of schistosome transmission ecology and snail population dynamics in natural waters. (author).

  3. Evaluation of [{sup 123}I]-CLINDE as a potent SPECT radiotracer to assess the degree of astroglia activation in cuprizone-induced neuroinflammation

    Energy Technology Data Exchange (ETDEWEB)

    Mattner, Filomena [Australian Nuclear Science and Technology Organisation, ANSTO LifeSciences, Sydney, NSW (Australia); Australia Nuclear Science and Technology Organisation, ANSTO LifeSciences, Kirrawee DC, NSW (Australia); Bandin, David Linares; Staykova, Maria; Fordham, Susan; Willenborg, David O. [Canberra Hospital, Neurosciences Research Unit (ANU Medical School), Woden, Canberra, ACT (Australia); Berghofer, Paula; Gregoire, Marie Claude; Ballantyne, Patrice; Quinlivan, Mitchell; Pham, Tien; Katsifis, Andrew [Australian Nuclear Science and Technology Organisation, ANSTO LifeSciences, Sydney, NSW (Australia)


    The purpose of this study was to assess the feasibility and sensitivity of the high-affinity translocator protein (TSPO) ligand [{sup 123}I]-CLINDE in imaging TSPO changes in vivo and characterise and compare astroglial and TSPO changes in the cuprizone model of demyelination and remyelination in C57BL/6 mice. C57BL/6 mice were fed with cuprizone for 4 weeks to induce demyelination followed by 2-4 weeks of standard diet (remyelination). Groups of mice were followed by in vivo single photon emission computed tomography (SPECT)/CT imaging using [{sup 123}I]-CLINDE and uptake correlated with biodistribution, autoradiography, immunohistochemistry, immunofluorescence and real-time polymerase chain reaction (RT-PCR). The uptake of [{sup 123}I]-CLINDE in the brain as measured by SPECT imaging over the course of treatment reflects the extent of the physiological response, with significant increases observed during demyelination followed by a decrease in uptake during remyelination. This was confirmed by autoradiography and biodistribution studies. A positive correlation between TSPO expression and astrogliosis was found and both activated astrocytes and microglial cells expressed TSPO. [{sup 123}I]-CLINDE uptake reflects astrogliosis in brain structures such as corpus callosum, caudate putamen, medium septum and olfactory tubercle as confirmed by both in vitro and in vivo results. The dynamics in the cuprizone-induced astroglial and TSPO changes, observed by SPECT imaging, were confirmed by immunofluorescence, RT-PCR and autoradiography. The highly specific TSPO radioiodinated ligand CLINDE can be used as an in vivo marker for early detection and monitoring of a variety of neuropathological conditions using noninvasive brain imaging techniques. (orig.)

  4. Alternative Radioligands for Investigating the Molecular Pharmacology of Melatonin Receptors. (United States)

    Legros, Céline; Brasseur, Chantal; Delagrange, Philippe; Ducrot, Pierre; Nosjean, Olivier; Boutin, Jean A


    Melatonin exerts a variety of physiologic activities that are mainly relayed through the melatonin receptors MT1 and MT2 Low expressions of these receptors in tissues have led to widespread experimental use of the agonist 2-[(125)I]-iodomelatonin as a substitute for melatonin. We describe three iodinated ligands: 2-(2-[(2-iodo-4,5-dimethoxyphenyl)methyl]-4,5-dimethoxy phenyl) (DIV880) and (2-iodo-N-2-[5-methoxy-2-(naphthalen-1-yl)-1H-pyrrolo[3,2-b]pyridine-3-yl])acetamide (S70254), which are specific ligands at MT2 receptors, and N-[2-(5-methoxy-1H-indol-3-yl)ethyl]iodoacetamide (SD6), an analog of 2-[(125)I]-iodomelatonin with slightly different characteristics. Here, we further characterized these new ligands with regards to their molecular pharmacology. We performed binding experiments, saturation assays, association/dissociation rate measurements, and autoradiography using sheep and rat tissues and recombinant cell lines. Our results showed that [(125)I]-S70254 is receptor, and can be used with both cells and tissue. This radioligand can be used in autoradiography. Similarly, DIV880, a partial agonist [43% of melatonin on guanosine 5'-3-O-(thio)triphosphate binding assay], selective for MT2, can be used as a tool to selectively describe the pharmacology of this receptor in tissue samples. The molecular pharmacology of both human melatonin receptors MT1 and MT2, using a series of 24 ligands at these receptors and the new radioligands, did not lead to noticeable variations in the profiles. For the first time, we described radiolabeled tools that are specific for one of the melatonin receptors (MT2). These tools are amenable to binding experiments and to autoradiography using sheep or rat tissues. These specific tools will permit better understanding of the role and implication in physiopathologic processes of the melatonin receptors.

  5. Further characterization of the low and high affinity binding components of the thyrotropin receptor. (United States)

    McQuade, R; Thomas, C G; Nayfeh, S N


    Following cross-linking with disuccinimidyl suberate and analysis by SDS-PAGE and autoradiography, both the high- and low-affinity TSH binding components exhibited two similar 125I-TSH-labeled bands, with Mr values of 80,000 and 68,000. IgG fractions from patients with Graves' disease inhibited 125I-TSH binding to both components, while normal IgG had no effect. Although not entirely conclusive, these results suggest that the high- and low-affinity components share similar subunit composition and antigenic determinants.

  6. Further characterization of the low and high affinity binding components of the thyrotropin receptor

    Energy Technology Data Exchange (ETDEWEB)

    McQuade, R.; Thomas, C.G. Jr.; Nayfeh, S.N.


    Following cross-linking with disuccinimdiyl suberate and analysis by SDS-PAGE and autoradiography, both the high- and low-affinity TSH binding components exhibited two similar /sup 125/I-TSH-labeled bands, with Mr values of 80,000 and 68,000. IgG fractions from patients with Graves' disease inhibited /sup 125/I-TSH binding to both components, while normal IgG had no effect. Although not entirely conclusive, these results suggest that the high- and low-affinity components share similar subunit composition and antigenic determinants.

  7. The fate of drotaverine-acephyllinate in rat and man. I. Absorption, distribution and excretion in the rat. (United States)

    Szatmári, I; Simon, G; Vargay, Z; Tóth, E; Szüts, T


    Two different labelled forms were used for the pharmacokinetic investigations: the carbon 1 in the isoquinoline ring (Drotaverine-14C-Acephyllinate) and the carboxyl group of theophylline-7-acetic acid (Drotaverine-Acephylline-14C-ate). Drotaverine-14C-Acephyllinate was rapidly absorbed from duodenal and ileal segments. Biliary excretion was substantial after oral administration and radioactivity was excreted mostly in the feces. Absorption of Drotavenine-Acephylline-14-C-ate from the gastrointestinal tract was very poor and radioactivity was therefore excreted for the most part in the feces. The results of the study were confirmed by whole body autoradiography.

  8. Investigation of Function of Novel Sperm Binding Protein HBRP in Human

    Institute of Scientific and Technical Information of China (English)


    Objective To investigate the biology function of novel protein related to bovie seminal plasma protein in human testis.Methods Recombination pcDNA3/HBRP was constructed and transfected to HEK293 cell and permanently expression cell line was established.The activity of protein kinase C (PKC) of the cell line was detected by autoradiography method.Results The stable expression cell line of HBRP was obtained.The HBRP inhibited the activity of PKC significantly.Conclusion One of the newfunctions of novel sperm binding protein in human is the inhibitor action on activity of PKC.It may be involved in the sperm capacitation,and acrosome reaction.

  9. Perfusion and metabolism imaging studies in Parkinson's disease

    DEFF Research Database (Denmark)

    Borghammer, Per


    Positron emission tomography (PET) and single photon emission computed tomography (SPECT) are important tools in the evaluation of brain blood flow and glucose metabolism in Parkinson's disease (PD). However, conflicting results are reported in the literature depending on the type of imaging data...... analysis employed. The present review gives a comprehensive summary of the perfusion and metabolism literature in the field of PD research, including quantitative PET studies, normalized PET and SPECT studies, autoradiography studies in animal models of PD, and simulation studies of PD data...

  10. Autoradiographic visualization of group III metabotropic glutamate receptors using [3H]-L-2-amino-4-phosphonobutyrate


    Hudtloff, Camilla; Thomsen, Christian


    In vitro receptor autoradiography using [3H]-L-2-amino-4-phosphonobutyrate ([3H]-L-AP4) binding to sections of rat brain has been characterized and shown to most likely represent labelling of group III metabotropic glutamate receptors.Specific [3H]-L-AP4 binding to rat brain sections was observed at high densities in the molecular layer of the cerebellar cortex and the outer layer of the superior colliculus. Moderate levels were observed throughout the cerebral cortex, in the molecular layer ...

  11. Astrocytosis precedes amyloid plaque deposition in Alzheimer APPswe transgenic mouse brain: a correlative positron emission tomography and in vitro imaging study

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez-Vieitez, Elena; Ni, Ruiqing; Voytenko, Larysa; Marutle, Amelia [Karolinska Institutet, Division of Translational Alzheimer Neurobiology, Centre for Alzheimer Research, Department of Neurobiology, Care Sciences and Society, Stockholm (Sweden); Gulyas, Balazs; Halldin, Christer [Karolinska Institutet, Centre for Psychiatric Research, Department of Clinical Neuroscience, Stockholm (Sweden); Nanyang Technological University, NTU - Imperial College, Lee Kong Chian School of Medicine, Singapore (Singapore); Toth, Miklos; Haeggkvist, Jenny [Karolinska Institutet, Centre for Psychiatric Research, Department of Clinical Neuroscience, Stockholm (Sweden); Nordberg, Agneta [Karolinska Institutet, Division of Translational Alzheimer Neurobiology, Centre for Alzheimer Research, Department of Neurobiology, Care Sciences and Society, Stockholm (Sweden); Karolinska University Hospital Huddinge, Department of Geriatric Medicine, Stockholm (Sweden)


    Pathological studies suggest that neuroinflammation is exacerbated by increased beta-amyloid (Aβ) levels in the brain early in Alzheimer's disease (AD). The time course and relationships between astrocytosis and Aβ deposition were examined using multitracer in vivo positron emission tomography (PET) imaging in an AD transgenic mouse model, followed by postmortem autoradiography and immunohistochemistry analysis. PET imaging with the amyloid plaque tracer {sup 11}C-AZD2184 and the astroglial tracer {sup 11}C-deuterium-L-deprenyl ({sup 11}C-DED) was carried out in APPswe mice aged 6, 8-15 and 18-24 months (4-6 animals/group) and in wild-type (wt) mice aged 8-15 and 18-24 months (3-6 animals/group). Tracer uptake was quantified by region of interest analysis using PMOD software and a 3-D digital mouse brain atlas. Postmortem brain tissues from the same APPswe and wt mice in all age groups were analysed for Aβ deposition and astrocytosis by in vitro autoradiography using {sup 3}H-AZD2184, {sup 3}H-Pittsburgh compound B (PIB) and {sup 3}H-L-deprenyl and immunostaining performed with antibodies for Aβ{sub 42} and glial fibrillary acidic protein (GFAP) in sagittal brain sections. {sup 11}C-AZD2184 PET retention in the cerebral cortices of APPswe mice was significantly higher at 18-24 months than in age-matched wt mice. Cortical and hippocampal {sup 11}C-DED PET binding was significantly higher at 6 months than at 8-15 months or 18-24 months in APPswe mice, and it was also higher than at 8-15 months in wt mice. In vitro autoradiography {sup 3}H-AZD2184 and {sup 3}H-PIB binding confirmed the in vivo findings with {sup 11}C-AZD2184 and demonstrated age-dependent increases in Aβ deposition in APPswe cortex and hippocampus. There were no significant differences between APPswe and wt mice in {sup 3}H-L-deprenyl autoradiography binding across age groups. Immunohistochemical quantification demonstrated more Aβ{sub 42} deposits in the cortex and hippocampus and more

  12. Experiments in nuclear science

    CERN Document Server

    Katz, Sidney A


    Characteristics of Geiger-Muller CountersResolving TimeBackground CorrectionsInverse Square LawCorrections for Geometry FactorsBack Scatter of RadiationCorrections for Self-absorptionRange of Beta RadiationsAbsorption of Beta RadiationAbsorption of Gamma RadiationRadioactive Decay and Instrument EfficiencyHalf-life DeterminationInvestigation of Two IndependentlyDecaying RadionuclidesHalf-life of a Long-lived RadionuclideAutoradiographyCalibration and Operation of the ElectroscopeProperties of Proportional CountersIntegral SpectraGamma Spectrometry IGamma Spectrometry IILiquid Scintillation Cou

  13. Cerebral markers of the serotonergic system in rat models of obesity and after Roux-en-Y gastric bypass

    DEFF Research Database (Denmark)

    Ratner, Cecilia; Ettrup, Anders; Bueter, Marco;


    Food intake and body weight are regulated by a complex system of neural and hormonal signals, of which the anorexigenic neurotransmitter serotonin (5-hydroxytryptamine or 5-HT) is central. In this study, rat models of obesity and weight loss intervention were compared with regard to several 5-HT...... markers. Using receptor autoradiography, brain regional-densities of the serotonin transporter (SERT) and the 5-HT(2A) and 5-HT(4) receptors were measured in (i) selectively bred polygenic diet-induced obese (pgDIO) rats, (ii) outbred DIO rats, and (iii) Roux-en-Y gastric bypass (RYGB)-operated rats. pg...

  14. Injected TFF1 and TFF3 bind to TFF2-immunoreactive cells in the gastrointestinal tract in rats

    DEFF Research Database (Denmark)

    Poulsen, S S; Thulesen, J; Hartmann, B


    and that injected TFF2 ends up in the mucus layer. In the present study, tissue binding and metabolism of parenterally administered human TFF1 and TFF3 in rats were described and compared to the immunohistochemical localization of the TFF peptides. 125I-TFF1 monomer and 125I-TFF3 mono- and dimer were given...... intravenously to female Wistar rats. The tissue distribution was assessed by gamma counting of organ samples and by autoradiography of histological sections. The degradation of 125I-TFF3 was studied by means of trichloracetic acid (TCA) precipitation and the saturability of the binding by administration...

  15. Distribution of pressure-induced fast axonal transport abnormalities in primate optic nerve. An autoradiographic study. (United States)

    Radius, R L


    The distribution of transport abnormalities in primate optic nerve from eyes subjected to five hours of pressure elevation (perfusion pressure of 35 mm Hg) was studied. Tissue autoradiography and electron microscopy were used to localize regions of the lamina cribrosa with increased transport interruption. A preferential involvement by this transport abnormality involved the superior, temporal, and inferior portions, to the exclusion of the nasal portion, of the optic nerve head. This observation supports the hypothesis that transport interruption seen in this model may be pertinent to the study of clinical glaucomatous neuropathy.

  16. Leukemia revisited

    Energy Technology Data Exchange (ETDEWEB)

    Cronkite, E P


    Selected features of the historical development of our knowledge of leukemia are discussed. The use of different methodologies for study of the nature of leukemic cell proliferation are analyzed. The differences between older cell kinetic data using tritiated thymidine and autoradiography and the newer cell culture methods are more apparent than real. It is suggested that tritiated thymidine and extracorporeal irradiation of the blood may be useful for therapeutic agents that have not been given an adequate trial. Radiation leukemogenesis presents an opportunity for study of the nature of leukemogenesis that has not been exploited adequately.

  17. Diurnal rhythm of melatonin binding in the rat suprachiasmatic nucleus

    Energy Technology Data Exchange (ETDEWEB)

    Laitinen, J.T.; Castren, E.; Vakkuri, O.; Saavedra, J.M.


    We used quantitative in vitro autoradiography to localize and characterize 2-/sup 125/I-melatonin binding sites in the rat suprachiasmatic nuclei in relation to pineal melatonin production. In a light:dark cycle of 12:12 h, binding density exhibited significant diurnal variation with a peak at the dark-light transition and a trough 12 hours later. Saturation studies suggested that the decreased binding at light-dark transition might be due to a shift of the putative melatonin receptor to a low affinity state.

  18. Question of the possible asymmetric polymerization of aspartic acid on kaolinite

    Energy Technology Data Exchange (ETDEWEB)

    McCullough, J.J.; Lemmon, R.M.


    McCullough and Lemmon have been unable to confirm the recent report that kaolinite catalyzes the polymerization of aqueous D- and L-aspartic acid at different rates. In experiments where DL-Asp was used, no induced optical rotation was found in the reaction solution. No evidence for polymer (or other product) formation was found when L-Asp-2-/sup 14/C was used, and products were searched by paper chromatography and x-ray film autoradiography. Asp is adsorbed by kaolinite, but no selectivity for one or the other enantiomer was observed.

  19. Replication of DNA during barley endosperm development

    DEFF Research Database (Denmark)

    Giese, H.


    The incorporation of [6-H-3]-thymidine into DNA of developing barley end sperm was examined by autoradiography of cross sections of seeds and DNA analysis. The majority of nuclear divisions took place in the very young endosperm, but as late as 25 days after anthesis there was evidence for DNA...... replication. The DNA content of the endosperm increases during development and in response to nitrogen application in parallel to the storage protein synthesis profile. The hordein genes were hypersensitive to DNase I treatment throughout development....

  20. Differential effect of NMDA and AMPA receptor blockade on protein synthesis in the rat infarct borderzone

    DEFF Research Database (Denmark)

    Christensen, Thomas; Bruhn, T; Frank, L


    treated with either saline, MK-801 (5 mg/kg i.p.) or NBQX (30 mg/kg i.p. x 3) were subjected to permanent MCAO. Regional CPSR and volumes of gray matter structures displaying normal CPSR were measured in coronal cryosections of the brain by quantitative autoradiography following an i.v. bolus injection....... Treatment with MK-801 significantly increased the volume of tissue with normal CPSR in the ischemic hemisphere compared to controls, whereas this was not seen with NBQX treatment. The results suggest that MK-801 and NBQX have different effects on peri-infarct protein synthesis after MCAO. Since both...

  1. Dynamic process of trace boron non-equilibrium grain boundary segregation and the effect of cooling rate

    Institute of Scientific and Technical Information of China (English)

    Ping Wu; Xinlai He; Bing Cao; Sen Chen


    The dynamic process of non-equilibrium grain boundary segregation of trace boron in Fe-40%Ni alloy during cooling andthe effect of cooling rate were investigated by boron tracking autoradiography technique. The results indicate that during coolingprocess, the amount of segregated boron on grain boundary firstly increases fast, then enters a comparatively even increasing stageand increases rapidly again at the third stage. The details of each stage varied with cooling rate are explained. When thc segregationdevelops to a certain degree, the segregated boron atoms transform fiom solute status to precipitate status.

  2. Theileria parva: effects of irradiation on a culture of parasitized bovine lymphoid cells. [Gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Irvin, A.D.; Brown, C.G.D.; Stagg, D.A.


    Aliquots of a culture of Theileria parva-infected bovine lymphoid cells were irradiated at 0, 300, 600, 900, and 1200 rads. The short-term effects of irradiation were evaluated on examination of Giemsa-stained smears and on autoradiography of cells labeled with (/sup 3/H)thymidine. Irradiation inhibited cell division but parasite division did not appear to be inhibited and macroschizont nuclear particles increased in number, frequently to several hundred per schizont. There was no evidence of an increased percentage switch from macro- to microschizont. Apparently viable cells were still present in all cultures 4 days after irradiation.

  3. Local cerebral glucose metabolism during controlled hypoxemia in rats. (United States)

    Pulsinelli, W A; Duffy, T E


    2-Deoxy-[14C]glucose metabolism was examined in brains of hypoxic, normotensive rats by autoradiography, which revealed alternating cortical columns of high and low metabolism. Activity in white matter was increased severalfold over that in adjacent gray matter. The columns were anatomically related to penetrating cortical arteries with areas between arteries demonstrating higher rates of metabolism. The results suggest the presence of interarterial tissue oxygen gradients that influence regional glucose metabolism. The relatively greater sensitivity of white matter metabolism to hypoxia may lead to an understanding of white matter damage in postanoxic leukoencephalopathy.

  4. [{sup 123}I]-3-Iodcytisin as possible radiotracer for the imaging of nicotinic acetylcholine receptors using single photon emission computer tomography; [{sup 123}I]-3-Iodcytisin als moeglicher Radiotracer fuer die Darstellung der nikotinergen Acetylcholin Rezeptoren mittels Single-Photon-Emissions-Computertomographie

    Energy Technology Data Exchange (ETDEWEB)

    Paulik, Dagmar Julia


    For the synthesis of [{sup 123}I]-3-Iodcytisin as possible radiotracer for the imaging of nicotinic acetylcholine (nACh) receptors using SPECT two different technologies were used: the radio-iodination with iodogen and the radio-iodination with nitric acid. The latter one showed higher efficiency. The radiotracer will allow to detect degenerative processes and other nACh-depending diseases in the brain (Alzheimer, Parkinson) and to observe the progress. The autoradiography is aimed to the imaging of the nACh receptors in the brain bypassing the brain-blood barrier. The highest activity was measured in the thalamus of mice and rat brains.

  5. Quality assurance and quantitative error analysis by tracer techniques

    Energy Technology Data Exchange (ETDEWEB)

    Schuetze, N.; Hermann, U.


    The locations, types and sources of casting defects have been tested by tracer techniques. Certain sites of moulds were labelled using /sup 199/Au, /sup 24/Na sodium carbonate solution, and technetium solution produced in the technetium generator on a /sup 99/Mo//sup 99/Tc elution column. Evaluations were made by means of activity measurements and autoradiography. The locations and causes of casting defects can be determined by error analysis. The surface defects of castings resulting from the moulding materials and from the blacking can be detected by technetium, the subsurface defects are located by gold.

  6. Radiotracers for PETT: new developments and perspectives

    Energy Technology Data Exchange (ETDEWEB)

    Fowler, J.S.; Wolf, A.P.


    Radiotracer development with positron emitters has its major focus on problems in the neurosciences. Progress is reviewed for high-level isotope production and labelled precurser synthesis with the medical cyclotron. The study of regional brain glucose metabolism represented the first extension of one of the methods of neurochemical autoradiography to humans and the study of brain protein synthesis and neurotransmitter receptors followed. In a more general sense, one PETT instrumentation will provide resolution in the 5 mm range is already emerging. Research status is reviewed. 103 references. (PSB)

  7. Cerebral 5-HT2A receptor binding, but not mGluR2, is increased in tryptophan hydroxylase 2 decrease-of-function mice

    DEFF Research Database (Denmark)

    Jørgensen, Christinna Vangsgaard; Jacobsen, Jacob P; Caron, Marc G


    interesting as a putative translational model of low endogenous 5-HT function that allows for assessment of adaptive changes in different anatomical regions. Here, we determined 5-HT2A receptor binding in several brain regions using in vitro receptor autoradiography and two different radioligands. When using...... show that in distinct anatomical regions, 5-HT2A receptor binding sites are up-regulated in 5-HT deficient mice, and this increase is not associated with changes in mGluR2 binding....

  8. Analyzing radioligand binding data. (United States)

    Motulsky, Harvey; Neubig, Richard


    Radioligand binding experiments are easy to perform, and provide useful data in many fields. They can be used to study receptor regulation, discover new drugs by screening for compounds that compete with high affinity for radioligand binding to a particular receptor, investigate receptor localization in different organs or regions using autoradiography, categorize receptor subtypes, and probe mechanisms of receptor signaling, via measurements of agonist binding and its regulation by ions, nucleotides, and other allosteric modulators. This unit reviews the theory of receptor binding and explains how to analyze experimental data. Since binding data are usually best analyzed using nonlinear regression, this unit also explains the principles of curve fitting with nonlinear regression.

  9. The oncogenic action of ionizing radiation on rat skin

    Energy Technology Data Exchange (ETDEWEB)

    Burns, F.J.


    Progress has occurred in several areas corresponding to the specific aims of the proposal: (1) Progression and multiple events in radiation carcinogenesis of rat skin as a function of LET; (2) cell cycle kinetics of irradiated rat epidermis as determined by double labeling and double emulsion autoradiography; (3) oncogene activation detected by in situ hybridization in radiation-induced rat skin tumors; (4) amplification of the c-myc oncogene in radiation-induced rat skin tumors as a function of LET; and (5) transformation of rat skin keratinocytes by ionizing radiation in combination with c-Ki-ras and c-myc oncogenes. 111 refs., 13 figs., 12 tabs.

  10. Comparison of two procedures for labelling the surface of the hydatid disease organism, Echinococcus granulosus, with /sup 125/I

    Energy Technology Data Exchange (ETDEWEB)

    McManus, D.P.; McLaren, D.J.; Clark, N.W.T.; Parkhouse, R.M.E.


    Living, intact protoscoleces of the British horse and sheep strains of Echinococcus granulosus were subjected to surface radioiodination procedures using /sup 125/I and Iodogen and /sup 125/I-Bolton Hunter reagent. Subsequent combined electron microscopy and autoradiography revealed specific surface membrane labelling with the Iodogen procedure, but significant tegumental labelling with the Bolton-Hunter reagent. The two parasite strains yielded different profiles of electrophoretically separated labelled proteins; the Iodogen method, not surprisingly, resulted in a less complex pattern of labelled polypeptides than the Bolton and Hunter reagent.

  11. Distribution of /sup 14/C-labelled ochratoxin A in pregnant mice

    Energy Technology Data Exchange (ETDEWEB)

    Appelgren, L.E.; Arora, R.G.


    Autoradiography was used to study the distribution of /sup 14/C-labelled ochratoxin A for up to 4 hr after its iv administration to mice at various stages of pregnancy. The highest /sup 14/C concentration was consistently found in the bile throughout the experimental period. The concentration of radioactivity in the tissues was found, in decreasing order, in the liver, kidney, blood, salivary glands, large vessels, brown fat, myocardium, uterus and lymphatic tissues. The toxin was shown to cross the placental barrier on day 9 of pregnancy, at which time it is most effective in producing fetal malformations.

  12. [{sup 18}F]p-MPPF: A Radiolabeled Antagonist for the Study of 5-HT{sub 1A} Receptors with PET

    Energy Technology Data Exchange (ETDEWEB)

    Plenevaux, A. E-mail:; Lemaire, C.; Aerts, J.; Lacan, G.; Rubins, D.; Melega, W.P.; Brihaye, C.; Degueldre, C.; Fuchs, S.; Salmon, E.; Maquet, P.; Laureys, S.; Damhaut, P.; Weissmann, D.; Le Bars, D.; Pujol, J.-F.; Luxen, A


    This paper summarizes the present status of the researches conducted with [{sup 18}F]4-(2'-methoxyphenyl)-1-[2'-[N-(2''-pyridinyl)-p-fluoro benzamido]ethyl] -piperazine known as [{sup 18}F]p-MPPF, a new 5-HT{sub 1A} antagonist for the study of the serotonergic neurotransmission with positron emission tomography (PET). This includes chemistry, radiochemistry, animal data (rats, cats, and monkeys) with autoradiography and PET, human data with PET, toxicity, and metabolism.

  13. Automatic analysis of intrinsic positional verification films brachytherapy using MATLAB; Analisis automatico de peliculas de verificacion posicional intrinsica en braqueterapia mediante MATLAB

    Energy Technology Data Exchange (ETDEWEB)

    Quiros Higueras, J. D.; Marco Blancas, N. de; Ruiz Rodriguez, J. C.


    One of the essential tests in quality control of brachytherapy equipment is verification auto load intrinsic positional radioactive source. A classic method for evaluation is the use of x-ray film and measuring the distance between the marks left by autoradiography of the source with respect to a reference. In our center has developed an automated method of measurement by the radiochromic film scanning and implementation of a macro developed in Matlab, in order to optimize time and reduce uncertainty in the measurement. The purpose of this paper is to describe the method developed, assess their uncertainty and quantify their advantages over the manual method. (Author)

  14. Increase in Blood-Brain Barrier Perrmeability, Oxidative Stress, and Activated Microglia in a Rat Model of Blast-Induced Traumatic Brain Injury (United States)


    3 X 3 min), followed by dipping in ict> cold water. The sections were exposed to Kodax BioMax Autoradiography Film (Kodak, Rochester, NY) for 55...days. All films were developed using a Kodak D-19 developer and analyzed in NIH Image v1.59 on a Power Macintosh connected to a Sony XC-77 CCD...matic brain injury in rats: an evJluation of sckcted measurcs. J Neuro- trauma 18:1207-1216. Hastings TG. 2tlO<J. The role of dopamine oxidation in

  15. The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos

    DEFF Research Database (Denmark)

    Østrup, Olga; Strejcek, F.; Petrovicova, I.;


    The aim of the present study was to investigate the role of RNA polymerase I (RPI) transcription in nucleolar development during major transcriptional activation (MTA) in cattle. Late eight-cell embryos were cultured in the absence (control group) or presence of actinomycin D (AD) (RPI inhibition......, Ad 0.2 µg/ml; total transcriptional inhibition, AD 2.0 µg/ml). Late four-cell embryos were cultured to late eight-cell stage in 0.2 µg/ml AD (MTA prevention, ADLT (long-term total transcriptional inhibition group). Embryos were processed for autoradiography, transmission electron microscopy...

  16. Rembrandt’s An Old Man in Military Costume: the underlying image re-examined


    Trentelman, K.; Janssens, K.; Van der Snickt, G.; Szafran, Y.; Woollett, A.T.; Dik, J.


    The painting An Old Man in Military Costume in the J. Paul Getty Museum, by Rembrandt Harmensz van Rijn, was studied using two complementary, element-specific imaging techniques—neutron activation autoradiography (NAAR) and macro-X-ray fluorescence (MA-XRF) mapping—to reveal the second, hidden painting. NAAR provided a strong image of the face and cloak of the underlying figure, along with an indication of the chemical composition. The single-element distribution maps produced by MA-XRF mappi...

  17. High expression of NPY receptors in the human testis. (United States)

    Körner, Meike; Waser, Beatriche; Thalmann, George N; Reubii, Jean Claude


    NPY receptors represent novel molecular therapeutic targets in cancer and obesity. However, the extent of NPY receptor expression in normal human tissues is poorly investigated. Based on the role of NPY in reproductive functions, the NPY receptor expression was studied in 25 normal human testes and, additionally, 24 testicular tumors using NPY receptor autoradiography. In the normal testis, Leydig cells strongly expressed NPY receptor subtype Y2, and small arterial blood vessels Y1. Y2 receptors were found to be functional with agonist-stimulated [(35)S]GTPγS binding autoradiography. Full functional integrity of the NPY system was further suggested by the immunohistochemical detection of NPY peptide in nerve fibers directly adjacent to Leydig cells and arteries. Germ cell tumors expressed Y1 and Y2 on tumor cells in 33% and Y1 on intratumoral blood vessels in 50%. Based on its strong NPY receptor expression in Leydig cells and blood vessels, the normal human testis represents a potentially important physiological and pharmalogical NPY target.

  18. Sulfur utilization of corals is enhanced by endosymbiotic algae

    Directory of Open Access Journals (Sweden)

    Ikuko Yuyama


    Full Text Available Sulfur-containing compounds are important components of all organisms, but few studies have explored sulfate utilization in corals. Our previous study found that the expression of a sulfur transporter (SLC26A11 was upregulated in the presence of Symbiodinium cells in juveniles of the reef-building coral Acropora tenuis. In this study, we performed autoradiography using 35S-labeled sulfate ions (35SO4 2− to examine the localization and amount of incorporated radioactive sulfate in the coral tissues and symbiotic algae. Incorporated 35SO4 2− was detected in symbiotic algal cells, nematocysts, ectodermal cells and calicoblast cells. The combined results of 35S autoradiography and Alcian Blue staining showed that incorporated 35S accumulated as sulfated glycosaminoglycans (GAGs in the ectodermal cell layer. We also compared the relative incorporation of 35SO4 2− into coral tissues and endosymbiotic algae, and their chemical fractions in dark versus light (photosynthetic conditions. The amount of sulfur compounds, such as GAGs and lipids, generated from 35SO4 2− was higher under photosynthetic conditions. Together with the upregulation of sulfate transporters by symbiosis, our results suggest that photosynthesis of algal endosymbionts contributes to the synthesis and utilization of sulfur compounds in corals.

  19. Autoradiographic study of the effects of pulsed electromagnetic fields on bone and cartilage growth in juvenile rats. (United States)

    Wilmot, J J; Chiego, D J; Carlson, D S; Hanks, C T; Moskwa, J J


    Application of pulsed electromagnetic fields (PEMF) has been used in growth and repair of non-union bone fractures. The similarities between the fibrocartilage callus in non-union bone fractures and the secondary cartilage in the mandibular condyle, both histologically and functionally, lead naturally to study the effects of PEMFs on growth in the condyle. The purposes of this study were: (1) to describe the effects of PEMFs on the growth of the condyle using autoradiography, [3H]-proline and [3H]-thymidine, and (2) to differentiate between the effects of the magnetic and electrical components of the field. Male pre-adolescent Sprague-Dawley rats (28 days old) were divided into three experimental groups of five animals each: (1) PEMF-magnetic (M), (2) PEMF-electrical (E) and (3) control, and were examined at three different times-3, 7 and 14 days of exposure. Each animal was exposed to the field for 8 h per day. Histological coronal sections were processed for quantitative autoradiography to determine the mitotic activity of the condylar cartilage and the amount of bone deposition. The PEMF (magnetic or electrical) had statistically significant effects only on the thickness of the articular zone, with the thickness in the PEMF-M group being the most reduced. Length of treatment was associated with predictable significant changes in the thickness of the condylar cartilage zones and the amount of bone deposition.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. Blood flow dependence of the intratumoral distribution of peripheral benzodiazepine receptor binding in intact mouse fibrosarcoma

    Energy Technology Data Exchange (ETDEWEB)

    Amitani, Misato [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan) and Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan)]. E-mail:; Zhang, Ming-Rong [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Noguchi, Junko [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); SHI Accelerator Service, Shinagawa-ku, Tokyo 141-8686 (Japan); Kumata, Katsushi [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Ito, Takehito [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); SHI Accelerator Service, Shinagawa-ku, Tokyo 141-8686 (Japan); Takai, Nobuhiko [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Suzuki, Kazutoshi [Radiochemistry Section, Department of Molecular Probes, Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Hosoi, Rie [Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan); Inoue, Osamu [Course of Allied Health Sciences, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871 (Japan)


    The intratumoral distribution of [{sup 11}C]AC-5216 binding, a novel peripheral benzodiazepine receptor (PBR) ligand, was examined by autoradiography both in vitro and in vivo using a murine fibrosarcoma model. The regional distribution of [{sup 11}C]AC-5216 in a tumor in vivo was significantly heterogeneous; the uptake of [{sup 11}C]AC-5216 was comparatively higher in the outer rim of the tumor and was lower in the central area. In contrast, the images obtained following the injection of [{sup 11}C]AC-5216 with a large amount of nonlabeled PK11195 showed a relatively homogeneous distribution, suggesting that [{sup 11}C]AC-5216 uptake represented specific binding to PBRs. In vitro autoradiograms of [{sup 11}C]AC-5216 binding were also obtained using the section of the fibrosarcoma that was the same as that used to examine in vivo binding. In vitro autoradiographic binding images showed homogeneous distribution, and significant discrepancies of the intratumoral distribution of [{sup 11}C]AC-5216 were observed between in vivo and in vitro images. The in vivo images of [{sup 11}C]AC-5216 uptake, compared with those of [{sup 14}C]iodoantipyrine uptake, obtained by dual autoradiography to evaluate the influence of blood flow revealed the similar intratumoral distributions of both tracers. These results indicate that the delivery process from the plasma to the tumor might be the rate-limiting step for the intratumoral distribution of PBR binding in vivo in a fibrosarcoma model.

  1. Molecular imaging of cholinergic processes in prostate cancer using {sup 11}C-donepezil and {sup 18}F-FEOBV

    Energy Technology Data Exchange (ETDEWEB)

    Stokholm, Morten Gersel; Bender, Dirk; Jakobsen, Steen; Froekiaer, Joergen; Borghammer, Per [Aarhus University Hospital, Department of Nuclear Medicine and PET Centre, Aarhus C (Denmark); Hoeyer, Soeren [Aarhus University Hospital, Department of Histopathology, Aarhus C (Denmark); Borre, Michael [Aarhus University Hospital, Department of Urology, Aarhus C (Denmark)


    High-grade prostate cancer (PC) displays parasympathetic neoneurogenesis. We investigated the binding of two PET tracers that visualize cholinergic nerves in PC tissue using autoradiography. Prostatectomy tissue was subjected to autoradiography with {sup 11}C-donepezil and {sup 18}F-FEOBV and correlated with Gleason scores (GS). Regions of interest on the autoradiograms were defined and quantified. Tracer binding in cancer tissue regions was compared with that in normal tissue. We included 13 patients with biopsy-verified PC. In particular, {sup 11}C-donepezil uptake was higher in ''high-grade'' PC (GS ≥4 + 3) than in ''low-grade'' PC and benign hyperplasia. {sup 11}C-donepezil uptake ranged from a mean of 56 % higher (GS 3 + 3) to 409 % higher (GS 4 + 4), and {sup 18}F-FEOBV uptake ranged from 67 % higher (GS 3 + 3) to 194 % higher (GS 4 + 5). The uptake of both tracers was higher in PC with a high GS than in PC with a low GS, but the difference was significant only for {sup 11}C-donepezil (p = 0.003). Uptake of PET tracers binding to cholinergic nerves was markedly higher in PC with a high GS than in PC with a low GS. This finding implies that {sup 11}C-donepezil PET/CT may be able to differentiate between low-grade and high-grade PC. (orig.)

  2. Assessment of [125I]WYE-230949 as a novel histamine H3 receptor radiopharmaceutical. (United States)

    Lewis, David Y; Champion, Sue; Wyper, David; Dewar, Deborah; Pimlott, Sally


    Histamine H3 receptor therapeutics have been proposed for several diseases such as schizophrenia, attention deficit hyperactivity disorder, Alzheimer's disease and obesity. We set out to evaluate the novel compound, [125I]WYE-230949, as a potential radionuclide imaging agent for the histamine H3 receptor in brain. [125I]WYE-230949 had a high in vitro affinity for the rat histamine H3 receptor (Kd of 6.9 nM). The regional distribution of [125I]WYE-230949 binding sites in rat brain, demonstrated by in vitro autoradiography, was consistent with the known distribution of the histamine H3 receptor. Rat brain uptake of intravenously injected [125I]WYE-230949 was low (0.11 %ID/g) and the ratio of specific: non-specific binding was less than 1.4, as determined by ex vivo autoradiography. In plasma, metabolism of [125I]WYE-230949 into a less lipophilic species occurred, such that less than 38% of the parent compound remained 30 minutes after injection. Brain uptake and metabolism of [125I]WYE-230949 were increased and specific binding was reduced in anaesthetised compared to conscious rats. [125I]WYE230949 is not a potential radiotracer for imaging rat histamine H3 receptors in vivo due to low brain uptake, in vivo metabolism of the parent compound and low specific binding.

  3. Assessment of [125I]WYE-230949 as a novel histamine H3 receptor radiopharmaceutical.

    Directory of Open Access Journals (Sweden)

    David Y Lewis

    Full Text Available Histamine H3 receptor therapeutics have been proposed for several diseases such as schizophrenia, attention deficit hyperactivity disorder, Alzheimer's disease and obesity. We set out to evaluate the novel compound, [125I]WYE-230949, as a potential radionuclide imaging agent for the histamine H3 receptor in brain. [125I]WYE-230949 had a high in vitro affinity for the rat histamine H3 receptor (Kd of 6.9 nM. The regional distribution of [125I]WYE-230949 binding sites in rat brain, demonstrated by in vitro autoradiography, was consistent with the known distribution of the histamine H3 receptor. Rat brain uptake of intravenously injected [125I]WYE-230949 was low (0.11 %ID/g and the ratio of specific: non-specific binding was less than 1.4, as determined by ex vivo autoradiography. In plasma, metabolism of [125I]WYE-230949 into a less lipophilic species occurred, such that less than 38% of the parent compound remained 30 minutes after injection. Brain uptake and metabolism of [125I]WYE-230949 were increased and specific binding was reduced in anaesthetised compared to conscious rats. [125I]WYE230949 is not a potential radiotracer for imaging rat histamine H3 receptors in vivo due to low brain uptake, in vivo metabolism of the parent compound and low specific binding.

  4. Radiolabeling and autoradiographic tracing of Toxocara canis larvae in male mice

    Energy Technology Data Exchange (ETDEWEB)

    Wade, S.E.; Georgi, J.R.


    Artificially hatched infective larvae of Toxocara canis were labeled with /sup 75/Se in Medium 199 (Gibco) containing /sup 75/Se-methionine. Male CD-1 mice were infected with radiolabeled larvae by intragastric intubation or by intraperitoneal injection. At intervals of 3-56 days mice were killed and the organs prepared for compressed organ autoradiography. Radioactivity of parasitic larvae showed an exponential decrease with time, reflecting catabolism of label with a biological half life of 26 days (effective half life of 21 days) making possible experiments lasting several months. Total body larva counts, estimated by total body autoradiography, displayed an overall downward trend, but the rate of reduction was probably not constant because no significant positive or negative trends were noted from day 14 onward in the numbers of larvae. The carcass accumulated the greatest number of larvae followed by the central nervous system, liver, and lung in that order. When the numbers of larvae were considered in relationship to the mass of tissue, there were 4 groupings: central nervous system, liver, lung, carcass, and kidney, and genito-urinary organ, pelt, and intestine. No significant difference between intragastric and intraperitoneal administration was observed in the larval distribution after the larvae had left the initial site of deposition.

  5. Methods in laboratory investigation. Autoradiographic demonstration of the specific binding and nuclear localization of 3H-dexamethasone in adult mouse lung. (United States)

    Beer, D G; Cunha, G R; Malkinson, A M


    This report describes the first autoradiographic demonstration of specific nuclear localization of 3H-dexamethasone in different cell types of the lung. Adult mouse lung tissue was incubated in vitro for 90 minutes with 17 nM 3H-dexamethasone in the presence or absence of various nonradioactive steroids. After extensive washing to remove any nonspecifically bound ligand, the specimens were processed for autoradiography using the thaw-mount method. In the absence of competing steroids, silver grains were localized in the nuclei of alveolar type II cells, bronchiolar and arteriolar smooth muscle cells, fibroblasts, and endothelial cells of the pulmonary vasculature. No significant nuclear concentration of label was observed in the bronchiolar epithelium, however. The specificity of 3H-dexamethasone labeling was demonstrated by incubating 17 nM 3H-dexamethasone with a 600-fold excess of either unlabeled dexamethasone, estrogen, dihydrotestosterone, or progesterone. These autoradiographic binding and steroid competition studies were confirmed by quantifying with liquid scintillation counting the specific 3H-dexamethasone binding in nuclear and cytosolic fractions prepared from lung tissues that had undergone identical incubation and washing procedures as those for autoradiography. These results demonstrate that many cell types in adult lung are targets for glucocorticoids and may respond to physiologic concentrations of this hormone.

  6. First preclinical evaluation of mono-[{sup 123}I]iodohypericin as a necrosis-avid tracer agent

    Energy Technology Data Exchange (ETDEWEB)

    Ni, Yicheng; Chen, Feng; Marchal, Guy [University Hospital Gasthuisberg, Department of Radiology, Leuven (Belgium); Huyghe, Dieter; Verbeke, Kristin; Verbruggen, Alfons M.; Bormans, Guy M. [University of Leuven, Laboratory of Radiopharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Leuven (Belgium); Witte, Peter A. de [University of Leuven, Laboratory of Pharmaceutical Biology and Phytopharmacology, Faculty of Pharmaceutical Sciences, Leuven (Belgium); Nuyts, Johan; Mortelmans, Luc [University Hospital Gasthuisberg, Department of Nuclear Medicine, Leuven (Belgium)


    We have labelled hypericin, a polyphenolic polycyclic quinone found in St. John's wort (Hypericum perforatum), with{sup 123}I and evaluated mono-[{sup 123}I]iodohypericin (MIH) as a potential necrosis-avid diagnostic tracer agent. MIH was prepared by an electrophilic radioiodination method. The new tracer agent was evaluated in animal models of liver infarction in the rat and heart infarction in the rabbit using single-photon emission computed tomography (SPECT), triphenyltetrazolium chloride (TTC) histochemical staining, serial sectional autoradiography and microscopy, and radioactivity counting techniques. Using in vivo SPECT imaging, hepatic and cardiac infarctions were persistently visualised as well-defined hot spots over 48 h. Preferential uptake of the tracer agent in necrotic tissue was confirmed by perfect match of images from post-mortem TTC staining, autoradiography (ARX) and histology. Radioactivity concentration in infarcted tissues was over 10 times (liver; 3.51% ID/g in necrotic tissue vs 0.38% ID/g in normal tissue at 60 h p.i.) and over 6 times (myocardium; 0.36% ID/g in necrotic tissue vs 0.054% ID/g in normal tissue; ratios up to 18 for selected parts on ARX images) higher than in normal tissues. The results suggest that hypericin derivatives may serve as powerful necrosis-avid diagnostic agents for assessment of tissue viability. (orig.)

  7. Visualization of early infarction in rat brain after ischemia using a translocator protein (18 kDa) PET ligand [11C]DAC with ultra-high specific activity. (United States)

    Yui, Joji; Hatori, Akiko; Kawamura, Kazunori; Yanamoto, Kazuhiko; Yamasaki, Tomoteru; Ogawa, Masanao; Yoshida, Yuichiro; Kumata, Katsushi; Fujinaga, Masayuki; Nengaki, Nobuki; Fukumura, Toshimitsu; Suzuki, Kazutoshi; Zhang, Ming-Rong


    The aim of this study was to visualize early infarction in the rat brain after ischemia using a translocator protein (TSPO) (18 kDa) PET ligand [(11)C]DAC with ultra-high specific activity (SA) of 3670-4450 GBq/μmol. An infarction model of rat brain was prepared by ischemic surgery and evaluated 2 days after ischemia using small-animal PET and in vitro autoradiography. Early infarction with a small increase of TSPO expression in the brain was visualized using PET with high SA [(11)C]DAC (average 4060 GBq/μmol), but was not distinguished clearly with usually reported SA [(11)C]DAC (37 GBq/μmol). Infarction in the rat brain 4 days after ischemia was visualized using high and usually reported SAs [(11)C]DAC. Displacement experiments with unlabeled TSPO-selective AC-5216 or PK11195 diminished the difference in radioactivity between ipsilateral and contralateral sides, confirming that the increased uptake on the infracted brain was specific to TSPO. In vitro autoradiography with high SA [(11)C]DAC showed that the TSPO expression increased on early infarction in the rat brain. High SA [(11)C]DAC is a useful and sensitive biomarker for the visualization of early infarction and the characterization of TSPO expression which was slightly elevated in the infarcted brain using PET.

  8. Glucose utilisation in the lungs of septic rats

    Energy Technology Data Exchange (ETDEWEB)

    Hansson, L.; Jeppsson, B. [Department of Surgery, University of Lund, Lund (Sweden); Ohlsson, T.; Sandell, A. [Department of Radiation Physics, University of Lund, Lund (Sweden); Valind, S. [Department of Clinical Physiology, University of Lund, Lund (Sweden); Luts, A. [Department of Physiology and Neuroscience, University of Lund, Lund (Sweden); Wollmer, P. [Department of Clinical Physiology, Malmoe University Hospital (Sweden)


    Sequestration and degranulation of leucocytes in the pulmonary microcirculation is considered to be a key event in the development of acute respiratory distress syndrome in patients with sepsis. Glucose serves as the main source of energy in activated leucocytes. The aim of this study was to assess whether glucose utilisation in the lungs can be used as an indicator of pulmonary leucocyte accumulation in an experimental model of sepsis of intra-abdominal origin. Sepsis was induced in rats by abdominal implantation of a gelatine capsule containing bacteria and rat colonic contents. Empty gelatine capsules were implanted in control animals. Animals were studied 6 and 12 h after sepsis induction. Glucose utilisation was measured as the tissue uptake of fluorine-18-fluorodeoxyglucose ({sup 18}FDG) 1 h after intravenous injection of the tracer. Micro-autoradiography was also performed after injection of tritiated deoxyglucose. We found increased uptake of {sup 18}FDG in the lungs of septic animals. The uptake also increased with time after sepsis induction. {sup 18}FDG uptake in circulating leucocytes was increased in septic animals compared with controls, and micro-autoradiography showed intense accumulation of deoxyglucose in leucocytes in the lungs of septic animals. We conclude that glucose utilisation is increased in the lungs of septic rats. Measurements of pulmonary glucose utilisation as an index of leucocyte metabolic activity may open new possibilities for studies of the pathophysiology of sepsis and for evaluation of therapeutic interventions. (orig.)

  9. Differentiation in boron distribution in adult male and female rats' normal brain: A BNCT approach

    Energy Technology Data Exchange (ETDEWEB)

    Goodarzi, Samereh, E-mail: [Department of Nuclear Engineering, Science and Research Branch, Islamic Azad University, PO Box 19395-1943, Tehran (Iran, Islamic Republic of); Pazirandeh, Ali, E-mail: [Department of Nuclear Engineering, Science and Research Branch, Islamic Azad University, PO Box 19395-1943, Tehran (Iran, Islamic Republic of); Jameie, Seyed Behnamedin, E-mail: [Basic Science Department, Faculty of Allied Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Baghban Khojasteh, Nasrin, E-mail: [Department of Nuclear Engineering, Science and Research Branch, Islamic Azad University, PO Box 19395-1943, Tehran (Iran, Islamic Republic of)


    Boron distribution in adult male and female rats' normal brain after boron carrier injection (0.005 g Boric Acid+0.005 g Borax+10 ml distilled water, pH: 7.4) was studied in this research. Coronal sections of control and trial animal tissue samples were irradiated with thermal neutrons. Using alpha autoradiography, significant differences in boron concentration were seen in forebrain, midbrain and hindbrain sections of male and female animal groups with the highest value, four hours after boron compound injection. - Highlights: Black-Right-Pointing-Pointer Boron distribution in male and female rats' normal brain was studied in this research. Black-Right-Pointing-Pointer Coronal sections of animal tissue samples were irradiated with thermal neutrons. Black-Right-Pointing-Pointer Alpha and Lithium tracks were counted using alpha autoradiography. Black-Right-Pointing-Pointer Different boron concentration was seen in brain sections of male and female rats. Black-Right-Pointing-Pointer The highest boron concentration was seen in 4 h after boron compound injection.

  10. A study of the aptitude of soils under natural conditions to retain radiostrontium; Etude de la vocation des sols en place a la retention du radiostrontium

    Energy Technology Data Exchange (ETDEWEB)

    Bovard, P.; Grauby, A. [Commissariat a l' Energie Atomique, Saclay (France).Centre d' Etudes Nucleaires


    Independently of the theoretical study of the propagation of radioactivity in the soil as a result of submersions or of radioactive rain, the authors have studied directly and practically how this radioactivity can vary in the actual soil. To this end a simple, rapid method has been perfected; it makes it possible to maintain for each soil sample the natural parameters (structure, humidity, etc.) without introducing boundary effects. In the laboratory, after charging the soil samples, part of the study of the propagation of radioactivity is done by autoradiography; finally, as a practical application, the study of an atomic site illustrates the methods described. (author) [French] Independamment de l'etude theorique de la propagation de la radioactivite dans le sol a la suite de submersions ou de pluies radioactives, les auteurs ont etudie directement et pratiquement comment pourrait evoluer cette radioactivite dans les sols en place. Pour cela, une methode simple et rapide a ete mise au point; elle permet de conserver pour chaque echantillon de sol, les parametres naturels (structure, humidite, etc...), sans introduire d'effets de paroi. En laboratoire, apres mise en charge des massifs preleves, une partie de l'etude de la propagation des radioelements est realisee par autoradiographie; enfin, une application pratique, l'etude d'un site atomique, illustre l'expose. (auteur)

  11. Distribution of /sup 109/Cd in the nervous system of rats after intravenous injection

    Energy Technology Data Exchange (ETDEWEB)

    Arvidson, B.; Tjaelve, H.


    The distribution of intravenously injected /sup 109/Cd in the nervous system was studied in rats twenty-four hr and one week after the injection. Measurements by gamma scintillation showed a high uptake of cadmium in peripheral sensory and autonomic ganglia whereas the uptake was low in the brain, cerebellum and spinal cord. The accumulation of cadmium in the sciatic nerve was significantly higher than in the brain and spinal nerve roots but lower than in ganglia. Autoradiography confirmed that there was no uptake of cadmium in the major part of the brain parenchyma, but showed an accumulation of the metal in areas outside the blood-brain barrier such as the hypophysis, meninges, choroid plexus and pineal gland. Within the peripheral nervous system, the autoradiography showed a localization of cadmium to dorsal root ganglia and, in addition, an uptake was seen in the connective tissue sheaths surrounding the sciatic nerve. The results indicate that the uptake of cadmium is correlated to regional variations in the permeability of blood vessels. The accumulation of cadmium in certain nervous structures may explain some of the neurotoxicological effects which have been demonstrated in animal experiments.

  12. Evaluation of Glucose Uptake in Normal and Cancer Cell Lines by Positron Emission Tomography. (United States)

    Maddalena, Francesca; Lettini, Giacomo; Gallicchio, Rosj; Sisinni, Lorenza; Simeon, Vittorio; Nardelli, Anna; Venetucci, Angela Assunta; Storto, Giovanni; Landriscina, Matteo


    To date, there is no definitive demonstration of the utility of positron emission tomography (PET) in studying glucose metabolism in cultured cell lines. Thus, this study was designed to compare PET to more standardized methods for the quantitative assessment of glucose uptake in nontransformed and transformed living cells and to validate PET for metabolic studies in vitro. Human colon and breast carcinoma cell lines and mouse embryo fibroblasts were evaluated for [(18)F]fluorodeoxyglucose ([(18)F]FDG) uptake by PET and autoradiography and 2-deoxyglucose (2-DG) incorporation by colorimetric assay and analyzed for the radiotoxic effects of [(18)F]FDG and the expression levels of glucose transporters. Indeed, [(18)F]FDG incorporation on PET was comparable to [(18)F]FDG uptake by autoradiography and 2-DG incorporation by colorimetric assay, although radiotracer-based methods exhibited more pronounced differences between individual cell lines. As expected, these data correlated with glucose transporters 1 to 4 and hexokinase II expression in tumor cell lines and mouse fibroblasts. Notably, [(18)F]FDG incorporation resulted in low apoptotic rates, with fibroblasts being slightly more sensitive to radiotracer-induced cell death. The quantitative analysis of [(18)F]FDG uptake in living cells by PET represents a valuable and reproducible method to study tumor cell metabolism in vitro, being representative of the differences in the molecular profile of normal and tumor cell lines.

  13. Stimulation of DNA synthesis in cultured rat alveolar type II cells

    Energy Technology Data Exchange (ETDEWEB)

    Leslie, C.C.; McCormick-Shannon, K.; Robinson, P.C.; Mason, R.J.


    Restoration of the alveolar epithelium after injury is thought to be dependent on the proliferation of alveolar type II cells. To understand the factors that may be involved in promoting type II cell proliferation in vivo, we determined the effect of potential mitogens and culture substrata on DNA synthesis in rat alveolar type II cells in primary culture. Type II cells cultured in basal medium containing 10% fetal bovine serum (FBS) exhibited essentially no DNA synthesis. Factors that stimulated /sup 3/H-thymidine incorporation included cholera toxin, epidermal growth factor, and rat serum. The greatest degree of stimulation was achieved by plating type II cells on an extracellular matrix prepared from bovine corneal endothelial cells and then by culturing the pneumocytes in medium containing rat serum, cholera toxin, insulin, and epidermal growth factor. Under conditions of stimulation of /sup 3/H-thymidine incorporation there was an increased DNA content per culture dish but no increase in cell number. The ability of various culture conditions to promote DNA synthesis in type II cells was verified by autoradiography. Type II cells were identified by the presence of cytoplasmic inclusions, which were visualized by tannic acid staining before autoradiography. These results demonstrate the importance of soluble factors and culture substratum in stimulating DNA synthesis in rat alveolar type II cells in primary culture.

  14. Human keratinocytes synthesize and secrete the extracellular matrix protein, thrombospondin. (United States)

    Wikner, N E; Dixit, V M; Frazier, W A; Clark, R A


    Thrombospondin (TSP) a glycoprotein originally identified as the endogenous lectin of platelets, is also synthesized by fibroblasts, endothelial cells, pneumocytes, smooth muscle cells, and macrophages. Thrombospondin is subdivided into functional domains which bind specifically to heparin, fibronectin, collagen, and to specific cellular receptors. It is found within the basement membranes of kidney, lung, smooth muscle, and skin. Thus TSP may serve as an important link between cells and matrices. Thrombospondin also has been reported at the epidermal-dermal junction. We wished to determine whether human keratinocytes synthesize and secrete TSP. Pure human keratinocytes were grown in defined medium without fibroblast feeder layers. Immunofluorescent staining with either rabbit polyclonal or mouse monoclonal antibodies to human platelet TSP yielded specific granular staining within the cytoplasm of keratinocytes. Culture media and cellular lysates were harvested from cultures metabolically labeled with [35S]methionine. Trichloroacetic acid precipitation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and autoradiography revealed a major labeled band comigrating with purified platelet TSP in both the media and the cellular lysates. Immunoprecipitation with either the polyclonal or the monoclonal anti-TSP antibodies followed by SDS-PAGE and autoradiography identified this band as TSP. Thus keratinocytes in culture synthesize and secrete TSP. Thrombospondin may play an important role in epidermal interactions with extracellular matrix.

  15. A radiopharmaceutical for imaging areas of lymphocytic infiltration: [sup 123]I-interleukin-2. Labelling procedure and animal studies

    Energy Technology Data Exchange (ETDEWEB)

    Signore, A.; Chianelli, M.; Toscano, A.; Monetini, L.; Ronga, G.; Pozzilli, P.; Negri, M. (Policlinico Umberto, Rome (Italy). Clinica Medica 2); Nimmon, C.C.; Britton, K.E. (Saint Bartholomew' s Hospital, London (United Kingdom). Dept. of Nuclear Medicine)


    The labelling of interleukin-2 (IL-2) with [sup 123]I and its in vivo application for imaging chronic pathological lymphocytic infiltrations are described. The lactoperoxidase/glucoseoxidase technique was the labelling method of choice leading to immunoreactive IL-2 with high specific activity. Labelled IL-2 was injected in diabetes-prone non-obese diabetic (NOD) mice with pancreatic lymphocytic infiltration. As control animals, Balb/c mice were used. As specificity control, monoclonal antibodies AMT13 and UCHT1, bovine serum albumin and [alpha]-lactalbumin were radioiodinated and injected in mice. Eighteen NOD mice and four control Balb/c mice were used for gamma camera imaging experiments. Fifty-four NOD and 20 Balb/c mice were used for time course single organ counting and autoradiography. Gamma camera images showed that radioactivity accumulated in the pancreatic region from the 10th minute onwards in NOD mice injected with [sup 123]I-IL-2 but not in Balb/c mice, or in NOD mice injected with control radiopharmaceuticals. These findings were confirmed by counting the radioactivity present in single organs. Autoradiography of NOD pancreas, after injection of labelled IL-2, showed that radioactivity was specifically associated with infiltrating lymphocytes. In conclusion, this technique is highly specific and easy to perform and we suggest its application in humans for in vivo detection of areas of lymphocytic infiltration. (Author).

  16. RPE cell surface proteins in normal and dystrophic rats

    Energy Technology Data Exchange (ETDEWEB)

    Clark, V.M.; Hall, M.O.


    Membrane-bound proteins in plasma membrane enriched fractions from cultured rat RPE were analyzed by two-dimensional gel electrophoresis. Membrane proteins were characterized on three increasingly specific levels. Total protein was visualized by silver staining. A maximum of 102 separate proteins were counted in silver-stained gels. Glycoproteins were labeled with 3H-glucosamine or 3H-fucose and detected by autoradiography. Thirty-eight fucose-labeled and 61-71 glucosamine-labeled proteins were identified. All of the fucose-labeled proteins were labeled with glucosamine-derived radioactivity. Proteins exposed at the cell surface were labeled by lactoperoxidase-catalyzed radioiodination prior to preparation of membranes for two-dimensional analysis. Forty separate 125I-labeled surface proteins were resolved by two-dimensional electrophoresis/autoradiography. Comparison with the glycoprotein map showed that a number of these surface labeled proteins were glycoproteins. Two-dimensional maps of total protein, fucose-labeled, and glucosamine-labeled glycoproteins, and 125I-labeled surface proteins of membranes from dystrophic (RCS rdy-p+) and normal (Long Evans or RCS rdy+p+) RPE were compared. No differences in the total protein or surface-labeled proteins were observed. However, the results suggest that a 183K glycoprotein is more heavily glycosylated with glucosamine and fucose in normal RPE membranes as compared to membranes from dystrophic RPE.

  17. Biodistribution, cellular localization, and in vivo tolerability of {sup 35}S-labeled antiinflammatory dendritic polyglycerol sulfate amine

    Energy Technology Data Exchange (ETDEWEB)

    Holzhausen, Cornelia [Freie Universität Berlin, Institute of Veterinary Pathology (Germany); Gröger, Dominic [Freie Universität Berlin, Institute of Chemistry and Biochemistry (Germany); Mundhenk, Lars [Freie Universität Berlin, Institute of Veterinary Pathology (Germany); Donat, Cornelius K. [Helmholtz-Zentrum Dresden-Rossendorf, Department of Neuroradiopharmaceuticals (Germany); Schnorr, Jörg [Charité-Universitätsmedizin Berlin, Institute of Radiology (Germany); Haag, Rainer [Freie Universität Berlin, Institute of Chemistry and Biochemistry (Germany); Gruber, Achim D., E-mail: [Freie Universität Berlin, Institute of Veterinary Pathology (Germany)


    Antiinflammatory dendritic polyglycerol sulfate (dPGS) holds great potential in the treatment and imaging of inflammatory processes. Here, we studied its biokinetic behavior, biodistribution, target cells, and in vivo toxicology. Following intravenous or subcutaneous application of {sup 35}sulfur-labeled dPGS amine with a molecular weight of 10.05 kDa and a hydrodynamic diameter of 5.7 ± 1.5 nm to mice, tissues were collected at specific time points (2, 15 min; 1, 24 h; 5, 21 days) and analyzed by liquid scintillation counting, autoradiography, radioluminography, and light microscopic autoradiography. The blood half-life of dPGS amine was 12 days. The major route of elimination was via the bile and feces. Elimination via the kidney and urine was only initially observed after i.v., but not after s.c. injection. Regardless of the administration mode, liver and spleen were late target organs where dPGS amine accumulated in phagocytic cells. Despite bioaccumulation, toxicological histopathology failed to identify any adverse effects at any time and in any tissues examined suggesting a high in vivo biocompatibility and encouraging future investigation for biomedical applications.

  18. Autoradiographic studies of synapsis formation on regenerated hair cells in the chich cochlea%小鸡再生听毛细胞及其突触连接的放射自显影

    Institute of Scientific and Technical Information of China (English)

    聂国辉; 汪吉宝


    目的采用光镜放射自显影(Light microscopic autoradiography, LM-ARG)及电镜放射自显影(Electron microscopic autoradiography, EM-ARG)技术,观察小鸡再生听毛细胞及其突触连接的特征。方法 7d龄纯种伊莎鸡20只,实验组(n=14),连续皮下注射庆大霉素10d;对照组(n=6)。皮下注射生理盐水,用药第4d开始注射氚标记胸腺嘧啶核苷,分别于停药当天及第7d取标本作LM-ARG和EM-ARG。结果停药当天组光镜下标记细胞多为支持细胞,位于损伤区基底膜上至表层之间;电镜下有标记的毛细胞位于腔面,银颗粒呈卷丝状,准确定位于细胞核上,毛细胞下可见尚未成熟的神经突触,内含较多囊泡及线粒体,突触特殊结构尚不明显。停药第7d时光镜发现受损区有标记的成熟毛细胞;电镜下再生毛细胞及突触特征与当天组相似。对照组未见标记的毛细胞和支持细胞。结论:庆大霉素中毒后鸡基底乳头中细胞增殖活跃,基底乳头中某些支持细胞作为前体细胞,通过有丝分裂增殖分化为毛细胞。再生毛细胞及其突触结构尚待进一步发育成熟。本结论为毛细胞再生及其神经突触的恢复提供了直接依据。%Objective To observe regenerated hair cells and new neural synapses in the chicken cochlea by the use of autoradiography. Methods Seven-day old chicks were given gentamicin or distilled water for 10 days. All birds were also injected with 3H-Thymidine (3H-TdR) begining on the fourth day of the period. Following postinjection survival periods of the same or 7 days, the basilar papilla was processed for light microscopic autoradiography (LM-ARG) and electron microscopic autoradiography (EM-ARG). Results Incorporation of 3H-TdR was seen over hair cells and supporting cells in the experimental papilla in regions hair cell loss. In EM-ARG, the silver grains had a simple structure, usually consisting of a single filment

  19. In-vivo imaging characteristics of two fluorinated flumazenil radiotracers in the rat

    Energy Technology Data Exchange (ETDEWEB)

    Dedeurwaerdere, Stefanie [University of Melbourne, Department of Medicine (RMH), Parkville, VIC (Australia); ANSTO, Radiopharmaceuticals Research Institute, Sydney (Australia); Gregoire, Marie-Claude; Fookes, Christopher; Greguric, Ivan; Pham, Tien; Loc' h, Christian; Katsifis, Andrew [ANSTO, Radiopharmaceuticals Research Institute, Sydney (Australia); Vivash, Lucy; O' Brien, Terence J.; Myers, Damian E. [University of Melbourne, Department of Medicine (RMH), Parkville, VIC (Australia); Roselt, Peter; Binns, David; Hicks, Rodney J. [Peter MacCallum Cancer Centre, Centre for Molecular Imaging, Melbourne (Australia)


    [{sup 11}C]Flumazenil shows promise as a clinical and research PET radiotracer to image changes in GABA{sub A} central benzodiazepine receptor (cBZR), but its widespread use has been limited by practical limitations of [{sup 11}C]. This study evaluated the imaging characteristics of two fluorinated PET radiotracers in rats in vivo: [{sup 18}F]fluoroflumazenil ([{sup 18}F]FFMZ) and [{sup 18}F]flumazenil ([{sup 18}F]FMZ). PET acquisitions were performed on a small-animal scanner following injection of [{sup 18}F]FFMZ in nine rats and [{sup 18}F]FMZ in eight rats. The following treatments were investigated: (1) injection of the tracer dose, (2) presaturation then injection of the tracer dose, and (3) injection of the tracer dose followed by a displacement injection. Unchanged tracer was measured in plasma and brain structures in four animals 10 and 30 min after injection, and ex-vivo autoradiography was also performed. For both [{sup 18}F]FFMZ and [{sup 18}F]FMZ maximal brain activity peaked rapidly, and was highest in the hippocampus (1.12{+-}0.06 SUV, 1.24{+-}0.10 SUV, respectively), and lowest in the pons (1.00{+-}0.07 SUV, 1.03{+-}0.09 SUV, respectively). By 50 min after injection, maximal uptake for [{sup 18}F]FFMZ and [{sup 18}F]FMZ had decreased in the hippocampus to 18{+-}3% and 80{+-}1% (p<0.01), respectively. The presaturation and displacement studies showed a higher nonspecific component for [{sup 18}F]FFMZ than for [{sup 18}F]FMZ. Metabolite studies showed that at 30 min only 10% of the signal was from [{sup 18}F]FFMZ in the brain. This nonspecific binding was apparent on autoradiography. In contrast, [{sup 18}F]FMZ accounted for >70% of the signal in the brain, which resulted in well-defined regional binding on autoradiography. These results demonstrate that [{sup 18}F]FMZ is a superior radiotracer to [{sup 18}F]FFMZ for in-vivo PET imaging of the GABA{sub A}/cBZR, having slower metabolism and leading to lower concentrations of metabolites in the brain

  20. Scintillation autoradiographic localization of 1,25-dihydroxyvitamin D/sub 3/ in chick intestine. [Tritium tracer techniques

    Energy Technology Data Exchange (ETDEWEB)

    Jones, P.G.; Haussler, M.R.


    The intracellular binding site of 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/) was determined via biochemical analysis of radioactive 1,25(OH)/sub 2/D/sub 3/ association with various chick tissues and then by direct autoradiography. When vitamin D-deficient chicks were injected intracardially with doses of tritiated 1,25(OH)/sub 2/D/sub 3/ and killed 2 h later, 2 to 3 times more radioactivity was found in the intestinal mucosa than was present in equal weights of pancreas, parathyroid, or liver tissue. Very little tritium was found in muscle tissue. The intestinally localized radioactivity was predominantly associated with the nuclear chromatin fraction, and binding of 1,25(OH)/sub 2/(/sup 3/H)D/sub 3/ to the nucleus was maximal 2 h after injection and at a dose of at least 0.52 nmol. Using this dose and time period, autoradiographic studies were done on duodenum and thoracic muscle of rachitic chicks injected with radioactive 1,25(OH)/sub 2/D/sub 3/ (11.2 Ci/mol). Thin sections of tissue were prepared for thaw and dry mount scintillation autoradiography as well as simple dip-coating autoradiography. After exposure for 4 to 6 months, a preferential concentration and retention of tritium-labeled 1,25(OH)/sub 2/D/sub 3/ was evident in the nuclei of intestinal villi and in the crypt of Lieberkuehn cells when each of the autoradiographic techniques was utilized. Quantitation of the labeled hormone confirms the significant nuclear accumulation in both villi and crypt cells. No such nuclear concentration of silver grains was observed in thoracic muscle cells, and the intestinal localization was abolished when a 100-fold excess of unlabeled 1,25(OH)/sub 2/D/sub 3/ was injected simultaneously with the radioactive hormone. It is concluded that 1,25(OH)/sub 2/D/sub 3/ is bound in a tissue-selective fashion to a high affinity, low capacity site within the nucleus of its intestinal target organ.

  1. In vivo and in vitro evidence that {sup 99m}Tc-HYNIC-interleukin-2 is able to detect T lymphocytes in vulnerable atherosclerotic plaques of the carotid artery

    Energy Technology Data Exchange (ETDEWEB)

    Glaudemans, Andor W.J.M.; Vries, Erik F.J. de; Koole, Michel; Luurtsema, Gert; Slart, Riemer H.J.A. [University Medical Center Groningen (Netherlands). Dept. of Nuclear Medicine and Molecular Imaging; Bonanno, Elena [Univ. of Rome Tor Vergata (Italy). Dept. of Anatomic Pathology; Galli, Filippo [Sapienza Univ, Rome (Italy). Nuclear Medicine Unit; Zeebregts, Clark J. [University Medical Center Groningen (Netherlands). Surgery (Div. Vascular Surgery); Boersma, Hendrikus H. [University Medical Center Groningen (Netherlands). Dept. of Nuclear Medicine and Molecular Imaging; University Medical Center Groningen (Netherlands). Clinical and Hospital Pharmacy; Taurino, Maurizio [Sapienza Univ., Rome (Italy). Vascular Surgery Unit; Signore, Alberto [University Medical Center Groningen (Netherlands). Dept. of Nuclear Medicine and Molecular Imaging; Sapienza Univ, Rome (Italy). Nuclear Medicine Unit


    Recent advances in basic science have established that inflammation plays a pivotal role in the pathogenesis of atherosclerosis. Inflammatory cells are thought to be responsible for the transformation of a stable plaque into a vulnerable one. Lymphocytes constitute at least 20 % of infiltrating cells in these vulnerable plaques. Therefore, the interleukin-2 (IL-2) receptor, being overexpressed on activated T lymphocytes, may represent an attractive biomarker for plaque vulnerability. The aim of this study was to evaluate the specificity of radiolabelled IL-2 [{sup 99m}Tc-hydrazinonicotinamide (HYNIC)-IL-2] for imaging the lymphocytic infiltration in carotid plaques in vivo by planar and single photon emission computed tomography (SPECT)/CT imaging and ex vivo by microSPECT and autoradiography. For the in vivo study, ten symptomatic patients with advanced plaques at ultrasound who were scheduled for carotid endarterectomy underwent {sup 99m}Tc-HYNIC-IL-2 scintigraphy. The images were analysed visually on planar and SPECT images and semi-quantitatively on SPECT images by calculating target to background (T/B) ratios. After endarterectomy, immunomorphological evaluation and immunophenotyping were performed on plaque slices. For the ex vivo studies, four additional patients were included and, after in vitro incubation of removed plaques with {sup 99m}Tc-HYNIC-IL-2, autoradiography was performed and microSPECT images were acquired. Visual analysis defined clear {sup 99m}Tc-HYNIC-IL-2 uptake in seven of the ten symptomatic plaques. SPECT/CT allowed visualization in eight of ten. A significant correlation was found between the number of CD25+ lymphocytes and the total number of CD25+ cells in the plaque and the T/B ratio with adjacent carotid artery as background (Pearson's r = 0.89, p = 0.003 and r = 0.87, p = 0.005, respectively). MicroSPECT imaging showed clear {sup 99m}Tc-HYNIC-IL-2 uptake within the plaque wall and not in the lipidic core. With autoradiography

  2. Putative melatonin receptors in a human biological clock

    Energy Technology Data Exchange (ETDEWEB)

    Reppert, S.M.; Weaver, D.R.; Rivkees, S.A.; Stopa, E.G.


    In vitro autoradiography with /sup 125/I-labeled melatonin was used to examine melatonin binding sites in human hypothalamus. Specific /sup 125/I-labeled melatonin binding was localized to the suprachiasmatic nuclei, the site of a putative biological clock, and was not apparent in other hypothalamic regions. Specific /sup 125/I-labeled melatonin binding was consistently found in the suprachiasmatic nuclei of hypothalami from adults and fetuses. Densitometric analysis of competition experiments with varying concentrations of melatonin showed monophasic competition curves, with comparable half-maximal inhibition values for the suprachiasmatic nuclei of adults (150 picomolar) and fetuses (110 picomolar). Micromolar concentrations of the melatonin agonist 6-chloromelatonin completely inhibited specific /sup 125/I-labeled melatonin binding, whereas the same concentrations of serotonin and norepinephrine caused only a partial reduction in specific binding. The results suggest that putative melatonin receptors are located in a human biological clock.

  3. Visualization of Functional Neuropeptide Y Receptors in the Mouse Hippocampus and Neocortex Using [35S]GTPγS Binding

    DEFF Research Database (Denmark)

    Elbrønd-Bek, Heidi; Gøtzsche, Casper René; Skinbjerg, Mette;


    The peptide transmitter neuropeptide Y (NPY) has been implicated in a plethora of actions in the central nervous system, including the hippocampus and neocortex (NeoCx). Previous studies using traditional receptor autoradiography show that NPY receptor binding is altered under various...... was further optimized to visualize the distribution of individual NPY receptor-mediated [35S]GTPγS binding in the mouse hippocampus and NeoCx using the endogenous ligand NPY in combination with optimized concentrations of selective antagonists for Y1, Y2, and Y5 NPY receptors. Consistent with previous studies......Cx. Furthermore, the effect of NPY receptor antagonists per se was also studied. Both BIIE0246 and L-152,804 significantly attenuated the basal [35S]GTPγS binding response, suggesting inverse agonism....

  4. GnRH receptors in human granulosa cells: Anatomical localization and characterization by autoradiographic study

    Energy Technology Data Exchange (ETDEWEB)

    Latouche, J.; Crumeyrolle-Arias, M.; Jordan, D.; Kopp, N.; Augendre-Ferrante, B.; Cedard, L.; Haour, F. (Institut Pasteur, Paris (France))


    The presence of receptors for GnRH in human ovary has been investigated by quantitative autoradiography. Simultaneous visualization and characterization of specific receptors on frozen sections were obtained on six pairs of human ovaries. Among them only one exhibited a large preovulatory follicle. This dominant follicle exhibited a specific and high affinity binding capacity for {sup 125}I-GnRHa exclusively localized on the granulosa cell layer. Analysis of saturation curve indicates a Kd value of 0.22 nM and Bmax of 9.6 fmol/mg protein. In contrast LH-hCG binding sites were present in all antral follicles. These data demonstrate for the first time the presence of high affinity GnRH receptors in human granulosa cells at a late stage of follicular maturation.

  5. L-(TH)glutamate binds to kainate-, NMDA- and AMPA-sensitive binding sites: an autoradiographic analysis

    Energy Technology Data Exchange (ETDEWEB)

    Monaghan, D.T.; Yao, D.; Cotman, C.W.


    The anatomical distribution of L-(TH)glutamate binding sites was determined in the presence of various glutamate analogues using quantitative autoradiography. The binding of L-(TH)glutamate is accounted for by the presence of 3 distinct binding sites when measured in the absence of CaS , Cl and Na ions. The anatomical distribution and pharmacological specificity of these binding sites correspond to that reported for the 3 excitatory amino acid binding sites selectively labelled by D-(TH)2-amino-5-phosphonopentanoate (D-(TH)AP5), (TH)kainate ((TH)KA) and (TH) -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ((TH)AMPA) which are thought to be selective ligands for the N-methyl-D-aspartate (NMDA), KA and quisqualate (QA) receptors, respectively. (Auth.). 29 refs.; 1 figure; 1 table.

  6. Absorption, distribution and elimination of drotaverine. (United States)

    Magyar, K; Lengyel, M; Knoll, J


    14C-drotaverine [1(3',4'-diethoxybenzale)-6, 7-diethoxy-1, 2,3, 4-tetrahydroisoquinoline. HCl; No-SpaR] is well absorbed after subcutaneous and oral administration in mice. Its distribution is not specific. After intravenous administration the drug penetrates rapidly into every organ as indicated by whole body autoradiography. In the first hours the concentration of drotaverine was higher in the intestinal wall than in the other tissues. The concentrations of drotaverine in the organs decrease soon after administration and the drug is excreted mainly with the bile as beta-glucuronide; 60% of the dose was in the bile collected during 5 hours. During 96 hours of observation, 67% of the radioactivity administered was found in the stools while only 20% of it was eliminated with urine.

  7. Isotopic labeling of mouse interferon by incorporation of radioactive amino acids during synthesis

    Energy Technology Data Exchange (ETDEWEB)

    DeMaeyer-Guignard, J.; Cachard, A.; DeMaeyer, E.


    Mouse interferon produced by C-243 cells induced with Newcastle disease virus was isotopically labeled by adding either (/sup 35/S)methionine or a /sup 14/C-labeled amino acid mixture to the culture medium. A method combining butyric acid and theophylline treatment and resulting in high interferon yields was used. Following purification by two-step affinity chromatography on poly(U) and antibody columns, the resulting material was analyzed on SDS-PAGE. The migration pattern of radioactivity and interferon coincided well and autoradiography revealed three major bands at migration distances corresponding, respectively, to 35, 28, and 22 K. Interferon represented 3.8% of all (/sup 35/S)methionine-labeled proteins and 2.6% of all /sup 14/C-amino acid-labeled proteins released into the medium.

  8. Corticosterone modulation of neurotransmitter receptors in rat hippocampus: a quantitative autoradiographic study

    Energy Technology Data Exchange (ETDEWEB)

    Biegon, A. (Hoffmann-La Roche, Inc., Nutley, NJ (USA). Dept. of Pharmacology); Rainbow, T.C. (Pennsylvania Univ., Philadelphia (USA). School of Medicine); McEwen, B.S. (Rockefeller Univ., New York (USA))


    The effect of adrenalectomy (ADX) and corticosterone (CORT) replacement on neurotransmitter receptors was studied in dorsal hippocampus of rat using quantitative autoradiography. ADX for one week causes an increase in (/sup 3/H)5-HT binding to 5-HT/sub 1/ receptors which is significant in the CA1 cell field. CORT treatment of ADX rats for 3-5 days results in localized reductions of (/sup 3/H)5-HT binding including a partial reversal of the increase observed after ADX in CA1. CORT treatment of ADX animals also decreases binding of (/sup 3/H)QNB to muscarinic receptors in the dorsal hippocampus, with a significant effect in an area designated as subiculum. No influence of CORT was detected on (/sup 3/H)prazosin binding to alpha/sub 1/ adrenergic receptors in dorsal hippocampus. Possible mechanisms for hormone effects on neurotransmitter receptor levels are discussed.


    Institute of Scientific and Technical Information of China (English)


    To investigate the expression of the receptors for vasoactive intestinal peptide (VIP) and secretin in colon cancer. Methods: This study visualized and characterized the receptors for VIP and secretin in the sequence of human tumor-free colon, adenoma, carcinoma, liver metastasis using storage phosphor autoradiography. Results: Receptors for VIP and secretin were demonstrated in tumor-free colon and colon tumors. A decrease in affinity of VIP receptors was shown in the colonic liver metastasis (Kd = 3.30 nmol) when compared with tumor-free colon (Kd = 0.82 nmol). An up-regulation of receptors for secretin was found in colonic liver metastases. Conclusions: VIP and secretin were both expressed on normal colon tissues. Binding of VIP decreased while secretin increased in colonic liver metastasis. A down-regulation of receptors for VIP in colonic liver metastases may helpful to understand the migration of colon cancer.

  10. Synthesis of [N-C[sup 3]H[sub 3

    Energy Technology Data Exchange (ETDEWEB)

    Wyrick, S.D.; Myers, A.M.; Booth, R.G.; Kula, N.S.; Baldessarini, R.J.; Mailman, R.B. (North Carolina Univ., Chapel Hill, NC (United States). School of Pharmacy and Brain and Development Research Center Harvard Medical School, Boston, MA (United States) Massachusetts General Hospital, Belmont, MA (United States). Mailman Research Center)


    Subsequent to the discovery that the (+)-benzomorphan sigma receptor ligands, (+)-pentazocine and (+)-N-allylnormetazocine, stimulated tyrosine hydroxylase activity and dopamine synthesis in rat striatum in vitro, we reported a similar effect on a structurally similar series of 1-phenyl-3-aminotetrahydronaphthalenes (phenylaminotetralins, PAT's). We previously reported the synthesis of tritium labeled Cl,OH-PAT to be used in radioreceptor and autoradiography studies and found that it labeled a sigma-like site in guinea pig brain with an apparent Kd of [approx] 50 pM and with a pharmacological profile unique from other known CNS receptors. Here we report the synthesis of high specific activity tritium labeled trans-(1R,3S)-(-)-H[sub 2]PAT as this enantiomer was found to be more active in the tyrosine hydroxylase assay and possessed approximately 45 fold greater affinity for the novel neuromodulatory sigma-like receptor. (author).

  11. Blockade of the MEK/ERK pathway with a raf inhibitor prevents activation of pro-inflammatory mediators in cerebral arteries and reduction in cerebral blood flow after subarachnoid hemorrhage in a rat model

    DEFF Research Database (Denmark)

    Maddahi, Aida; Ansar, Saema; Chen, Qingwen


    hours, cerebral arteries were harvested, and iNOS, interleukin (IL)-6, IL-1ß, matrix metalloproteinase (MMP)-9, tissue inhibitors of metalloproteinase (TIMP)-1, and phosphorylated ERK1/2 were investigated by immunofluorescence, real-time polymerase chain reaction (PCR), and Western blot analysis....... Cerebral blood flow (CBF) was measured using autoradiography. Protein levels of MMP-9, TIMP-1, iNOS, IL-6, and IL-1ß were increased after SAH, as were mRNA levels of IL-6, MMP-9, and TIMP-1. After SAH, pERK1/2 was increased, but CBF was reduced. Treatment with SB-386023-b at 0 or 6 hours after SAH...

  12. Blockade of the MEK/ERK pathway with a raf inhibitor prevents activation of pro-inflammatory mediators in cerebral arteries and reduction in cerebral blood flow after subarachnoid hemorrhage in a rat model

    DEFF Research Database (Denmark)

    Maddahi, Aida; Ansar, Saema; Chen, Qingwen


    hours, cerebral arteries were harvested, and iNOS, interleukin (IL)-6, IL-1β, matrix metalloproteinase (MMP)-9, tissue inhibitors of metalloproteinase (TIMP)-1, and phosphorylated ERK1/2 were investigated by immunofluorescence, real-time polymerase chain reaction (PCR), and Western blot analysis....... Cerebral blood flow (CBF) was measured using autoradiography. Protein levels of MMP-9, TIMP-1, iNOS, IL-6, and IL-1β were increased after SAH, as were mRNA levels of IL-6, MMP-9, and TIMP-1. After SAH, pERK1/2 was increased, but CBF was reduced. Treatment with SB-386023-b at 0 or 6 hours after SAH...

  13. A nuclear method to authenticate Buddha images (United States)

    Khaweerat, S.; Ratanatongchai, W.; Channuie, J.; Wonglee, S.; Picha, R.; Promping, J.; Silva, K.; Liamsuwan, T.


    The value of Buddha images in Thailand varies dramatically depending on authentication and provenance. In general, people use their individual skills to make the justification which frequently leads to obscurity, deception and illegal activities. Here, we propose two non-destructive techniques of neutron radiography (NR) and neutron activation autoradiography (NAAR) to reveal respectively structural and elemental profiles of small Buddha images. For NR, a thermal neutron flux of 105 n cm-2s-1 was applied. NAAR needed a higher neutron flux of 1012 n cm-2 s-1 to activate the samples. Results from NR and NAAR revealed unique characteristic of the samples. Similarity of the profile played a key role in the classification of the samples. The results provided visual evidence to enhance the reliability of authenticity approval. The method can be further developed for routine practice which impact thousands of customers in Thailand.

  14. Induction of DNA repair synthesis by ultraviolet radiation and methylmethanesulphonate in cultured mouse lymphocytes

    Energy Technology Data Exchange (ETDEWEB)

    Bianchi, V.; Zantedeschi, A.; Levis, A.G. (Padua Univ. (Italy). Ist. di Zoologia e Anatomia Comparata); Ronchese, F. (Inst. of Pathological Anatomy, Padua (Italy))


    The induction of DNA repair synthesis by UV-radiation and methylmethanesulphonate (MMS) was studied in mouse lymphocytes and leukemic cells by means of autoradiography and scintillation counting, after labelling in vitro with tritiated thymidine ((/sup 3/H)dThd). Repair stimulation was detected by both procedures in LSTRA AND YC8 leukemic cell lines as well as in primary fibroblasts of BALB/c and BALB/Mo mice. No stimulation was observed in primary cultures of lymphocytes from the spleen, thymus and lymph-nodes of the same mice. In primary lymphocytes neither stimulation with concanavalin A (Con A) nor pre-incubation with 5-bromodeoxyuridine (BUdR) were effective in making evident DNA repair. The data put into question the reliability of the repair test for the prediction of carcinogenic potential of chemicals.

  15. Zinc Blotting Assay for Detection of Zinc-Binding Prolamin in Barley (Hordeum vulgare) Grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Langkilde, Ane; Vincze, Éva


    In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol-soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. 65Zn overlay and blotting techniques have been widely used for detecting zinc......-binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (65ZnCl2) blotting technique was optimized to detect zinc-binding prolamins, followed by development of an easy-to-follow nonradioactive colorimetric...... zinc blotting method with a zinc-sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS-PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc-binding specificity of certain proteins was detected either by autoradiography or color...

  16. Effect of intracarotid injection of iopamidol on local cerebral glucose utilization in rat brain. (United States)

    d'Avella, D; Cicciarello, R; Albiero, F; Piscitelli, G; Fiori, M G; Mesiti, M; Princi, P; d'Aquino, S


    We assessed, by means of the [14C]-2-deoxy-D-glucose autoradiography method, the effect of intracarotid injection of a nonionic, low-osmolar contrast medium (iopamidol) on local cerebral glucose utilization in the rat brain. Contrast medium was injected at 20 degrees C and at 37 degrees C, and the relative changes in local cerebral glucose utilization were measured. At 20 degrees C the viscosity of the contrast agent was about twice that of the same solution at 37 degrees C, and resulted in a statistically significant increase in local cerebral glucose utilization in the hemisphere ipsilateral to the side of intracarotid infusion. Saline control studies showed that the metabolic change was not related to either the solution temperature or the osmolality. These findings suggest that increased viscosity of a contrast medium may contribute to its neurotoxic effects during cerebral angiography, hence emphasizing the importance of preheating contrast material to avoid adverse reactions.

  17. Parametric mapping of 5HT1A receptor sites in the human brain with the Hypotime method: theory and normal values

    DEFF Research Database (Denmark)

    Møller, Mette; Rodell, Anders; Gjedde, Albert


    analysis. METHODS: A total of 19 healthy volunteers (age range, 23-73 y) underwent PET to test the Hypotime application of the chemical microsphere properties of (11)C-WAY to identify regions of binding and nonbinding on the exclusive basis of the rate of washout of (11)C-WAY. RESULTS: The results...... of the Hypotime method were compared with the simplified but multilinearized reference tissue method (MLSRTM). The distribution of receptor BP(ND) obtained with Hypotime was consistent with previous autoradiography of postmortem brain tissue, with the highest values of BP(ND) recorded in the medial temporal lobe...... and decline of receptor availability with age. The values in the basal ganglia and cerebellum were negligible. The MLSRTM, in contrast, yielded lower BP(ND) in all regions and only weakly revealed the decline with age. CONCLUSION: The simple and computationally efficient Hypotime method gave reliable values...

  18. Effects of unilateral 6-OHDA lesions on [3H]-N-propylnorapomorphine binding in striatum ex vivo and vulnerability to amphetamine-evoked dopamine release in rat

    DEFF Research Database (Denmark)

    Palner, Mikael; Kjaerby, Celia; Knudsen, Gitte M


    ligands should likewise be fitter than antagonists for detecting responses to denervation in positron emission tomography studies of idiopathic Parkinson's disease. Agonist binding increases in vivo are likely to reflect the composite of a sensitization-like phenomenon, and relatively less competition...... to be more vulnerable to competition from endogenous dopamine than was the antagonist ligand [(11)C]raclopride, measured ex vivo in mouse striatum, and subsequently in multi-tracer PET studies of analogous design. Based on these results, we predicted that prolonged dopamine depletion would result...... in a preferential increase in agonist binding, and a lesser competition from residual dopamine to the agonist binding. To test this hypothesis we used autoradiography to measure [(3)H]NPA and [(3)H]raclopride binding sites in hemi-parkinsonian rats with unilateral 6-OHDA lesions, with and without amphetamine...

  19. Analytical, ultrastructural, autoradiographic and biochemical studies on [3H]dicarboxylic acid added to cultures of melanoma cells. (United States)

    Ward, B J; Breathnach, A S; Robins, E J; Bhasin, Y; Ethridge, L; Passi, S; Nazzaro-Porro, M


    Lentigo maligna and malignant melanoma can be treated by dicarboxylic acids (C9 and C12), which are competitive inhibitors of tyrosinase. We therefore studied the intracellular location and possible sites of action of dodecanedioic acid (C12) in murine melanoma cells, using EM autoradiography and biochemical analysis of lipid extracts by HPLC. Significant levels of radioactivity were found in the mitochondria and in the nuclei but not in association with membranes of rough endoplasmic reticulum, Golgi-associated endoplasmic reticulum, or Golgi apparatus, and not in coated vesicles or melanosomes. Biochemical analysis revealed that the diacid underwent beta-oxidation, which occurs only in mitochondria. The results suggest that the toxicity of dicarboxylic acids in melanoma cells is not related to anti-tyrosinase activity but may be due to interference with oxidoreductase enzymes in the mitochondria and possibly to inhibition of DNA synthesis in the nucleus.

  20. Physical properties, structure, and shape of radioactive Cs from the Fukushima Daiichi Nuclear Power Plant accident derived from soil, bamboo and shiitake mushroom measurements. (United States)

    Niimura, Nobuo; Kikuchi, Kenji; Tuyen, Ninh Duc; Komatsuzaki, Masakazu; Motohashi, Yoshinobu


    We conducted an elution experiment with contaminated soils using various aqueous reagent solutions and autoradiography measurements of contaminated bamboo shoots and shiitake mushrooms to determine the physical and chemical characteristics of radioactive Cs from the Fukushima Daiichi Nuclear Power Plant accident. Based on our study results and data in the literature, we conclude that the active Cs emitted by the accident fell to the ground as granular non-ionic materials. Therefore, they were not adsorbed or trapped by minerals in the soil, but instead physically adhere to the rough surfaces of the soil mineral particles. Granular Cs* can be transferred among media, such as soils and plants. The physical properties and dynamic behavior of the granular Cs* is expected to be helpful in considering methods for decontamination of soil, litter, and other media.

  1. Temporal variation of cesium isotope concentrations and atom ratios in zooplankton in the Pacific off the east coast of Japan (United States)

    Ikenoue, Takahito; Takata, Hyoe; Kusakabe, Masashi; Kudo, Natsumi; Hasegawa, Kazuyuki; Ishimaru, Takashi


    After the Fukushima Daiichi Nuclear Power Plant accident in March 2011, concentrations of cesium isotopes (133Cs, 134Cs, and 137Cs) were measured in zooplankton collected in the Pacific off the east coast of Japan from May 2012 to February 2015. The time series of the data exhibited sporadic 137Cs concentration peaks in zooplankton. In addition, the atom ratio of 137Cs/133Cs in zooplankton was consistently high compared to that in ambient seawater throughout the sampling period. These phenomena cannot be explained fully by the bioaccumulation of 137Cs in zooplankton via ambient seawater intake, the inclusion of resuspended sediment in the plankton sample, or the taxonomic composition of the plankton. Autoradiography revealed highly radioactive particles within zooplankton samples, which could be the main factor underlying the sporadic appearance of high 137Cs concentrations in zooplankton as well as the higher ratio of 137Cs/133Cs in zooplankton than in seawater.

  2. Prenatal neurogenesis in the telencephalon of the precocial mouse Acomys cahirinus. (United States)

    Brunjes, P C; Korol, D L; Stern, K G


    [3H]thymidine autoradiography was employed to examine the times of formation of the major neuronal classes in the forebrain of the precocial mouse Acomys cahirinus. Dams received 3 thymidine injections over a 24 h period on either embryonic Day 14, 18, 20, 22, 29 or 36. Age-related changes in the distribution and number of heavily labeled cells were noted. Acomys exhibited later onset and more protracted periods of cell generation than the phylogenetically related, altricial laboratory rat or mouse, indicating considerable differences in patterns of early growth between the species. Understanding the factors responsible for these differences could lend important insights into evolutionary mechanisms involved in the process of speciation.

  3. Repeated administration of alpha7 nicotinic acetylcholine receptor (nAChR) agonists, but not positive allosteric modulators, increases alpha7 nAChR levels in the brain

    DEFF Research Database (Denmark)

    Christensen, Ditte Z; Mikkelsen, Jens D; Hansen, Henrik H;


    -induced phosphorylation of Erk2 in the prefrontal cortex occurs following acute, but not repeated administration. Our results demonstrate that repeated agonist administration increases the number of alpha7 nAChRs in the brain, and leads to coupling versus uncoupling of specific intracellular signaling....... Here we investigate the effects of repeated agonism on alpha7 nAChR receptor levels and responsiveness in vivo in rats. Using [(125)I]-alpha-bungarotoxin (BTX) autoradiography we show that acute or repeated administration with the selective alpha7 nAChR agonist A-582941 increases the number of alpha7 n......-120596 and NS1738 do not increase [(125)I]-BTX binding. Furthermore, A-582941-induced increase in Arc and c-fos mRNA expression in the prefrontal cortex is enhanced and unaltered, respectively, after repeated administration, demonstrating that the alpha7 nAChRs remain responsive. Contrarily, A-582941...

  4. Biodistribution of [sup 125]I labeled monoclonal antibody against gamma seminoprotein in the nude mice bearing human benign prostatic hyperplasia xenografts

    Energy Technology Data Exchange (ETDEWEB)

    Fujino, Awato (Kitasato Univ., Sagamihara, Kanagawa (Japan). School of Medicine)


    The biodistribution of [sup 125]I labeled monoclonal antibody against gamma seminoprotein ([gamma]-Sm) in the nude mice bearing human benign prostatic hyperplasia xenograft was evaluated by whole body autoradiography and by counting of radioactivity in organs. The monoclonal antibody (murine Ig G[sub 1], K) to [gamma]-Sm which was established in this institute and its F(ab')[sub 2] fragment were radioiodinated using Iodogen method. The autoradiograms demonstrated specific uptake of [sup 125]I-intact Ig G as well as [sup 125]I-F(ab')[sub 2] within the prostatic adenoma xenografts 4 days after intravenous administrations. Radioactivity in the xenograft was relatively higher than those in the liver, kidney or lung. These results suggest that radio-labeled monoclonal antibody against [gamma]-Sm might be applicable for radioimmunodetection of prostatic tumors which produce [gamma]-Sm. (author).

  5. DEPFET--a pixel device with integrated amplification

    CERN Document Server

    Neeser, W; Buchholz, P; Fischer, P; Holl, P; Kemmer, J; Klein, P; Koch, H; Löcker, M; Lutz, Gerhard; Matthäy, H; Strüder, L; Trimpl, M; Ulrici, J; Wermes, N


    In the DEPFET pixel concept, the absorbed radiation directly modulates the channel current of a p-JFET transistor being integrated into a fully depleted high ohmic silicon substrate in every pixel cell, offering very low noise operation at room temperature. Hence, DEPFET pixels open new possibilities in biomedical applications, but also have a potential in particle physics and astrophysics. Second prototype 50 mu mx50 mu m single pixels as well as large (64x64) DEPFET matrices have been successfully produced and operated confirming the low noise behavior (12e). Device studies as well as a full DEPFET pixel Bioscope system to be used in real-time digital autoradiography with excellent spatial and energy resolution for X-rays are presented.

  6. Muscarinic M3 receptor subtype gene expression in the human heart. (United States)

    Hellgren, I; Mustafa, A; Riazi, M; Suliman, I; Sylvén, C; Adem, A


    The heart is an important target organ for cholinergic function. In this study, muscarinic receptor subtype(s) in the human heart were determined using reverse transcription-polymerase chain reaction. Our results demonstrated muscarinic receptor M2 and M3 subtype RNA in left/right atria/ventricles of donor hearts. Receptor autoradiography analysis using selective muscarinic ligands indicated an absence of M1 receptor subtype in the human heart. The level of muscarinic receptor binding in atria was two to three times greater than in ventricles. Our results suggest that muscarinic receptors in the human heart are of the M2 and M3 subtypes. This is the first report of M3 receptors in the human myocardium.

  7. Autoradiographic analysis of tritiated imipramine binding in the human brain post mortem: effects of suicide

    Energy Technology Data Exchange (ETDEWEB)

    Gross-Isseroff, R.; Israeli, M.; Biegon, A.


    In vitro quantitative autoradiography of high-affinity tritiated imipramine binding sites was performed on brains of 12 suicide victims and 12 matched controls. Region-specific differences in imipramine binding were found between the two groups. Thus, the pyramidal and molecular layers of the cornu ammoni hippocampal fields and the hilus of the dentate gyrus exhibited 80%, 60%, and 90% increases in binding in the suicide group, respectively. The postcentral cortical gyrus, insular cortex, and claustrum had 45%, 28%, and 75% decreases in binding in the suicide group, respectively. No difference in imipramine binding was observed in prefrontal cortical regions, in the basal ganglia, and in mesencephalic nuclei. No sex and postmortem delay effects on imipramine binding were found. Imipramine binding was positively correlated with age, the effect of age being most pronounced in portions of the basal ganglia and temporal cortex.

  8. Site of iodination in rat mammary gland

    Energy Technology Data Exchange (ETDEWEB)

    Strum, J.M.


    The ability of the mammary gland to take up and organically bind radioiodide was studied in non-pregnant, pregnant, and lactating rats. Autoradiography was used to determine whether duct cells or alveolar cells are responsible for iodination in the rat mammary gland. Iodination was not detected in mammary glands from non-pregnant rats, but occurred late in the twelfth day of gestation and continued throughout pregnancy and lactation. Protein-containing vacuoles in alveolar cells and casein-like proteins in milk were the major sites where iodination occurred within the gland. Milk proteins in the lumens of ductules adjacent to alveoli were also iodinated. In contrast, ducts, myoepithelial cells, fat cells, blood vessels and other histological components of the gland did not show iodinating capability. Cytochemistry was also used to identify endogenous mammary peroxidase activity in the same glands, and it was found that the presence and location of this enzyme were correlated with the ability to iodinate.

  9. Effects of compound rehmannia formula on dopamine transporter content in the corpus striatum of Parkinson's disease rats treated with levodopa

    Institute of Scientific and Technical Information of China (English)

    Ruijng Luo; Jiancheng He


    Long-term application of levodopa (L-3, 4-dihydroxyphenylalanine, L-DOPA) for Parkinson's disease can lead to adverse effects and reduce the amount of dopamine transporter (DAT) in the corpus striatum. The present study attempted to verify whether increasing the amount of DAT can reduce the adverse effects of L-DOPA. The specific radioactive uptake value of DAT in the corpus striatum of the lesioned hemisphere was significantly decreased, but was significantly increased following administration of compound rehmannia formula [Radix rehmanniae preparata (prepared rehmannia root), Concha margaritifera usta (nacre), Radix paeoniae alba (white peony alba), Radix salviae miltiorrhizae (Danshen root), Scorpio (scorpion), green tea] for 4 weeks. The changes in DAT 125I-beta-carbomethoxy-3 beta-(4-iodophenyl) tropane autoradiography were consistent with those in radioactivity. The results revealed that the compound rehmannia formula can reduce the adverse effects of L-DOPA in treating Parkinson′s disease, possibly by increasing the amount of DAT.

  10. Monoclonal antibody raised against human mitotic cyclin B1, identifies cyclin B-like mitotic proteins in synchronized onion (Allium cepa L.) root meristem. (United States)

    Chaudhuri, S K; Ghosh, S


    Cyclin B-like mitotic proteins have been detected in synchronized Allium cepa L. root tip cells by using mouse monoclonal anti-cyclin B1 antibody raised against human cyclin B1. Immunoblot shows two closely placed isoforms of cyclin B-like proteins having an apparent molecular weight around 54 kDa. In vivo [35S]-methionine labelling followed by immunoprecipitation and autoradiography indicates that cyclin B-like proteins are mainly synthesized in the G2 phase of the cell cycle and destroyed in late mitosis. Immunoblotting data depict that the level of cyclin B-like proteins reaches the maximum at the late G2 to early M phase; and it becomes degraded in the late hours of mitosis. Moreover, the cyclin B isoforms are stabilized in colchicine-arrested metaphase cells as already reported in animal cells.

  11. [From automation to robotics]. (United States)


    The introduction of automation into the laboratory of biology seems to be unavoidable. But at which cost, if it is necessary to purchase a new machine for every new application? Fortunately the same image processing techniques, belonging to a theoretic framework called Mathematical Morphology, may be used in visual inspection tasks, both in car industry and in the biology lab. Since the market for industrial robotics applications is much higher than the market of biomedical applications, the price of image processing devices drops, and becomes sometimes less than the price of a complete microscope equipment. The power of the image processing methods of Mathematical Morphology will be illustrated by various examples, as automatic silver grain counting in autoradiography, determination of HLA genotype, electrophoretic gels analysis, automatic screening of cervical smears... Thus several heterogeneous applications may share the same image processing device, provided there is a separate and devoted work station for each of them.

  12. Hydrogen diffusion and distribution in alloy 600 and related effects on the plasticity; Diffusion et distribution de l`hydrogene dans l`alliage 600, effet sur la plasticite

    Energy Technology Data Exchange (ETDEWEB)

    Lecoester, F.; Brass, A.M.; Chene, J. [Laboratoire de metalurgie structurale, URA CNRS No. 1107, Bat. 413, Universit Paris-Sud, 91405 Orsay (France); Noel, D. [Electricite de France (EDF), 77 - Ecuelles (France)


    Hydrogen can play a part in several mechanisms proposed for explaining the stress corrosion cracking of nickel based alloy 600, used in steam generators of pressurized water nuclear reactors. This study presents data on diffusion and hydrogen trapping in alloy 600 as well as the embrittlement which results from it. Distribution data were obtained by deuterium analysis of samples cathodically charged with heavy water. Secondary ion mass spectrometry, liquid scintillation counting and tritium autoradiography have been used for analysis. Data on hydrogen embrittlement were obtained by imposed tensile tests on samples with or without cathodic charging. Different microstructures were studied. The results show that alloy 600 embrittlement greatly depend on the structure and increases with the degree of intergranular precipitation. An effect of hydrogen on the plasticity of the alloy was noted. (author). 9 refs.

  13. Differences in human skin between the epidermal growth factor receptor distribution detected by EGF binding and monoclonal antibody recognition

    DEFF Research Database (Denmark)

    Green, M R; Couchman, J R


    , the eccrine sweat glands, capillary system, and the hair follicle outer root sheath, generally similar in pattern to that previously reported for full-thickness rat skin and human epidermis. The same areas also bound EGF-R1 but in addition the monoclonal antibody recognized a cone of melanin containing......, indicating that, at least for SVK14 cells, EGF-R1 binding provides a reliable marker for EGF binding. Explanations for the discrepancies between these two methods for determining EGF receptor distribution in human skin are discussed, including the possibility that latent EGF receptors, unable to bind [125I......Two methods have been used to examine epidermal growth factor (EGF) receptor distribution in human scalp and foreskin. The first employed [125I]EGF viable explants and autoradiography to determine the EGF binding pattern while the second used a monoclonal antibody to the human EGF receptor to map...

  14. Rapid DNA sequencing by horizontal ultrathin gel electrophoresis. (United States)

    Brumley, R L; Smith, L M


    A horizontal polyacrylamide gel electrophoresis apparatus has been developed that decreases the time required to separate the DNA fragments produced in enzymatic sequencing reactions. The configuration of this apparatus and the use of circulating coolant directly under the glass plates result in heat exchange that is approximately nine times more efficient than passive thermal transfer methods commonly used. Bubble-free gels as thin as 25 microns can be routinely cast on this device. The application to these ultrathin gels of electric fields up to 250 volts/cm permits the rapid separation of multiple DNA sequencing reactions in parallel. When used in conjunction with 32P-based autoradiography, the DNA bands appear substantially sharper than those obtained in conventional electrophoresis. This increased sharpness permits shorter autoradiographic exposure times and longer sequence reads. Images PMID:1870968

  15. Autoradiographic imaging of the serotonin transporter, using S-[{sup 18}F](fluoromethyl)-(+)-McN5652 ([{sup 18}F]Me-McN) in the brains of several animal species

    Energy Technology Data Exchange (ETDEWEB)

    Kretzschmar, M.; Zessin, J.; Brust, P.; Cumming, P. [PET Centre of Aarhus Univ. Hospitals, Aarhus C (Denmark); Bergmann, R.


    The [{sup 18}F]fluoromethyl analogue of (+)-McN5652 ([{sup 18}F]Me-McN) was recently proposed as a new potential PET tracer [1]. To further validate its use in PET, we studied the binding of [{sup 18}F]Me-McN in the brains of rats and pigs using autoradiography. The binding was compared with the uptake of the known 5-HT uptake inhibitor [{sup 3}H] citalopram [2] and the radioligand (+)-[{sup 11}C]McN5652. The binding of the three compounds was qualitatively identical in the autoradiograms of the individual brains. Intense labelling was observed in regions known to be serotonin uptake sites. The binding was specifically inhibited, using the 5-HT uptake inhibitors citalopram and fluoxetine. (orig.)

  16. Real-time monitoring of DNAzyme cleavage process using fluorescent assay

    Institute of Scientific and Technical Information of China (English)

    Xiang Xian Meng; Xiao Hai Yang; Ke Min Wang; Wei Hong Tan; Qiu Ping Guo


    Detection of deoxyribozyme (DNAzyme) cleavage process usually needs complex and time-consuming radial labeling, gel electrophoresis and autoradiography. This paper reported an approach to detect DNAzyme cleavage process in real time using a fluorescence probe. The probe was employed as DNAzyme substrate to convert directly the cleavage information into fluorescence signal in real time. Compared with traditional approach, this non-isotope method not only brought a convenient means to monitor the DNAzyme cleavage reaction, but also offered abundant dynamic data for choosing potential gene therapeutic agents. It provides a new tool for DNAzyme research, as well as a new insight into research on human disease diagnosis. Based on this method, 8-17deoxyribozyme (8-17DNAzyme) against hepatitis C virus RNA (HCV-RNA) was designed and the cleavage process was studied in real time.

  17. An approach for serotonin depletion in pigs: effects on serotonin receptor binding

    DEFF Research Database (Denmark)

    Ettrup, Anders; Kornum, Birgitte R; Weikop, Pia


    concentrations of 5-HT in seven distinct brain structures from one hemisphere: frontal and occipital cortex, striatum, hippocampus, cerebellum, rostral, and caudal brain stem, were determined. The other hemisphere was processed for receptor autoradiography. Treatments with 50 mg/kg and 100 mg/kg pCPA caused...... is increasingly used as an experimental animal model especially in neuroscience research. Here, we present an approach for serotonin depletion in the pig brain. Central serotonin depletion in Danish Landrace pigs was achieved following 4 days treatment with para-chlorophenylalanine (pCPA). On day 5, tissue...... average decreases in 5-HT concentrations of 61% ± 14% and 66% ± 16%, respectively, and a substantial loss of 5-HT immunostaining was seen throughout the brain. The serotonin depletion significantly increased 5-HT₄ receptor binding in nucleus accumbens, but did not alter 5-HT(1A) and 5-HT(2A) receptor...

  18. Changes in Binding of [(123)I]CLINDE, a High-Affinity Translocator Protein 18 kDa (TSPO) Selective Radioligand in a Rat Model of Traumatic Brain Injury

    DEFF Research Database (Denmark)

    Donat, Cornelius K; Gaber, Khaled; Meixensberger, Jürgen


    After traumatic brain injury (TBI), secondary injuries develop, including neuroinflammatory processes that contribute to long-lasting impairments. These secondary injuries represent potential targets for treatment and diagnostics. The translocator protein 18 kDa (TSPO) is expressed in activated...... microglia cells and upregulated in response to brain injury and therefore a potential biomarker of the neuroinflammatory processes. Second-generation radioligands of TSPO, such as [(123)I]CLINDE, have a higher signal-to-noise ratio as the prototype ligand PK11195. [(123)I]CLINDE has been employed in human...... and sacrificed at 6, 24, 72 h and 28 days post surgery. TSPO expression was assessed in brain sections employing [(123)I]CLINDE in vitro autoradiography. From 24 h to 28 days post surgery, injured animals exhibited a marked and time-dependent increase in [(123)I]CLINDE binding in the ipsilateral motor...

  19. An approach for serotonin depletion in pigs: effects on serotonin receptor binding

    DEFF Research Database (Denmark)

    Ettrup, Anders; Kornum, Birgitte R; Weikop, Pia


    concentrations of 5-HT in seven distinct brain structures from one hemisphere: frontal and occipital cortex, striatum, hippocampus, cerebellum, rostral, and caudal brain stem, were determined. The other hemisphere was processed for receptor autoradiography. Treatments with 50 mg/kg and 100 mg/kg pCPA caused...... is increasingly used as an experimental animal model especially in neuroscience research. Here, we present an approach for serotonin depletion in the pig brain. Central serotonin depletion in Danish Landrace pigs was achieved following 4 days treatment with para-chlorophenylalanine (pCPA). On day 5, tissue...... average decreases in 5-HT concentrations of 61% ± 14% and 66% ± 16%, respectively, and a substantial loss of 5-HT immunostaining was seen throughout the brain. The serotonin depletion significantly increased 5-HT4 receptor binding in nucleus accumbens, but did not alter 5-HT(1A) and 5-HT(2A) receptor...

  20. Film analysis systems and applications

    Energy Technology Data Exchange (ETDEWEB)

    Yonekura, Y.; Brill, A.B.


    The different components that can be used in modern film analysis systems are reviewed. TV camera and charge-coupled device sensors coupled to computers provide low cost systems for applications such as those described. The autoradiography (ARG) method provides an important tool for medical research and is especially useful for the development of new radiopharmaceutical compounds. Biodistribution information is needed for estimation of radiation dose, and for interpretation of the significance of observed patterns. The need for such precise information is heightened when one seeks to elucidate physiological principles/factors in normal and experimental models of disease. The poor spatial resolution achieved with current PET-imaging systems limits the information on radioreceptor mapping, neutrotransmitter, and neuroleptic drug distribution that can be achieved from patient studies. The artful use of ARG in carefully-controlled animal studies will be required to provide the additional information needed to fully understand results obtained with this new important research tool. (ERB)

  1. Differential immediate and sustained memory enhancing effects of alpha7 nicotinic receptor agonists and allosteric modulators in rats

    DEFF Research Database (Denmark)

    Thomsen, Morten Skøtt; El-Sayed, Mona; Mikkelsen, Jens D


    . Subsequent [(125)I]-bungarotoxin autoradiography revealed no direct correlation between α7 nAChR levels in frontal cortical or hippocampal brain regions and short-term memory with either compound. Additionally, repeated treatment with A-582941 did not affect mRNA expression of RIC-3 or the lynx-like gene...... products lynx1, lynx2, PSCA, or Ly6H, which are known to affect nAChR function. In conclusion, both α7 nAChR agonists and PAMs exhibit sustained pro-cognitive effects after repeated administration, and altered levels of the α7 nAChR per se, or that of endogenous regulators of nAChR function, are likely...

  2. Direct demonstration of insulin receptor internalization. A quantitative electron microscopic study of covalently bound /sup 125/I-photoreactive insulin incubated with isolated hepatocytes

    Energy Technology Data Exchange (ETDEWEB)

    Gorden, P.; Carpentier, J.L.; Moule, M.L.; Yip, C.C.; Orci, L.


    When /sup 125/I-insulin is incubated with isolated rodent hepatocytes at 37 degrees C, the ligand initially binds to the plasma membrane of the cell and is subsequently internalized by adsorptive endocytosis. To confirm directly that the insulin receptor is internalized with the ligand, we covalently linked photoreactive /sup 125/I-N sigma B29 (azidobenzoyl) insulin to its specific hepatocyte receptor and followed its fate by quantitative electron microscopic autoradiography. We found that the covalently linked photoreactive insulin is internalized by the cell in fashion analogous to the internalization of ordinary /sup 125/I-insulin, indicating that, at least under these conditions, the insulin receptor is internalized with the ligand.

  3. Filter transfer of genomic libraries in a state accessible to DNA-binding proteins. (United States)

    Beebee, T J


    I have developed a method for transferring plaque DNA of lambda genomic libraries onto 3MM filters in a state accessible to DNA-binding proteins. DNA bound to 3MM is available to proteins as large as Escherichia coli RNA polymerase and maintains template activity similar to that in free solution. Lambda Plaques can be lifted onto 3MM filter disks, deproteinized, and used for transcription assays in vitro. The RNA synthesized is complementary to phage rather than to E. coli DNA and plaques can be identified by autoradiography. Furthermore, the filters can subsequently be probed with radioactive nucleic acids under standard hybridization conditions. Finally, colorimetric assays can be employed with lactate dehydrogenase (LDH) A in which plaques are identified by the localized reduction of nitroblue tetrazolium.

  4. Effect of dislocation configuration on non-equilibrium boron segregation during cooling

    Institute of Scientific and Technical Information of China (English)


    Different densities and configurations of crystal defects were obtained in an austenitic Fe-30%Ni alloy and an ultra low carbon bainitic (ULCB) alloy by undergoing different deformations and annealing treatments at high temperatures. Boron segregation on grain boundaries and subgrain boundaries during air-cooling were revealed by means of the particle tracking autoradiography technique. It is found that non-equilibrium segregation is resisted in deformed grains after recovery and polygonization, boron-depleted zones seem to be quite clear in recrystallized grains than those in deformed original grains during cooling. Subgrain boundaries and polygonized dislocation cells have a significant effect on non-equilibrium boron segregation during the air-cooling. The results implicates that dislocation configuration is a more important factor affecting boron segregation at grain boundaries rather than the density of defects itself in the grain.

  5. Synthesis of heat shock proteins in rat brain cortex after transient ischemia. (United States)

    Dienel, G A; Kiessling, M; Jacewicz, M; Pulsinelli, W A


    Cell-free protein synthesis and two-dimensional gel autoradiography were used to characterize early postischemic protein synthesis in rat neocortex. Severe forebrain ischemia was induced for 30 min (four-vessel occlusion model) and followed by 3 h of recirculation. Polysomes were isolated from the cerebral cortex, translated in vitro in a reticulocyte system, and analyzed by two-dimensional gel electrophoresis. The translation products of postischemic polysomes included a major new protein family (70 kDa) with multiple isoelectric variants that was found to comigrate with the 68- to 70-kDa "heat shock" protein synthesized from polysomes of hyperthermic rats. Two other stress proteins (93 and 110 kDa) also appeared to be synthesized in increased amounts after ischemia. A complement of proteins that was indistinguishable from that of controls was also synthesized after ischemia, indicating that messenger ribonucleic acid coding for most brain proteins is preserved after ischemia and is bound to polysomes.

  6. Evidence for xylitol 5-P production in human dental plaque

    Energy Technology Data Exchange (ETDEWEB)

    Waaler, S.M. (Department of Preclinical Techniques and Material Sciences and Department of Pedodontics, Dental Faculty, University of Oslo, Oslo (Norway))


    The Turku sugar studies indicated that xylitol may possess a caries-therapeutic effect. More recent data show that xylotol exhibits a bacteriostatic activity on a wide range of bacteria based on uptake and expulsion of xylitol. Intracellular xylitol 5-P appears to be a key substance associated with inhibition of bacterial metabolism by xylitol. This has been shown in studies with pure strains of bacteria, mainly Streptococcus mutans. The aim of the present study was to examine if production of xylitol 5-P occurs in freshly collected dental plaque which is exposed to labeled xylitol. Plaque extracts were analyzed by thin-layer chromatography combined with autoradiography and high performance liquid chromatography. Strong indications were obtained that xylitol 5-P is readily produced by dental plaque. No other significant xylitol metabolites were identified. The bacteriostatic properties of xylitol in plaque are a mechanism which may well account for the caries-therapeutic effect of xylitol. (au).

  7. Regional Myocardial Substrate Uptake in Hypertensive Rats: A Quantitative Autoradiographic Measurement (United States)

    Yonekura, Yoshiharu; Bertrand Brill, A.; Som, Prantika; Yamamoto, Kazutaka; Srivastava, Suresh C.; Iwai, Junichi; Elmaleh, David R.; Livni, Eli; Strauss, H. William; Goodman, Mark M.; Knapp, Furn F.


    Severe hypertension causes global and regional changes in myocardial perfusion and substrate utilization. Regional perfusion and fatty acid utilization were evaluated by dual-tracer autoradiography in normotensive and hypertensive rats of the Dahl strain. The regional distributions of perfusion and fatty acid utilization were homogeneous in normotensive rats. Severe hypertension was associated with a homogeneous pattern of regional perfusion, but fatty acid utilization was focally decreased in the free wall of the left ventricle. The decrease in fatty acid uptake was associated with a concomitant increase in glucose utilization. These findings suggest that severe hypertension is associated with uniform myocardial perfusion and focal alterations in the substrates used for the performance of myocardial work.

  8. [Studies of biologic activation associated with molecular receptor increase and tumor response in ChL6/L6 protocol patients; Studies in phantoms; Quantitative SPECT; Preclinical studies; and Clinical studies]. DOE annual report, 1994--95

    Energy Technology Data Exchange (ETDEWEB)

    DeNardo, S.J.


    The authors describe results which have not yet been published from their associated studies listed in the title. For the first, they discuss Lym-1 single chain genetically engineered molecules, analysis of molecular genetic coded messages to enhance tumor response, and human dosimetry and therapeutic human use radiopharmaceuticals. Studies in phantoms includes a discussion of planar image quantitation, counts coincidence correction, organ studies, tumor studies, and {sup 90}Y quantitation with Bremsstrahlung imaging. The study on SPECT discusses attenuation correction and scatter correction. Preclinical studies investigated uptake of {sup 90}Y-BrE-3 in mice using autoradiography. Clinical studies discuss image quantitation verses counts from biopsy samples, S factors for radiation dose calculation, {sup 67}Cu imaging studies for lymphoma cancer, and {sup 111}In MoAb imaging studies for breast cancer to predict {sup 90}Y MoAb therapy.

  9. Comparison of US and FRG post-irradiation examination procedures to measure statistically significant failure fractions of irradiated coated-particle fuels. [HTGR

    Energy Technology Data Exchange (ETDEWEB)

    Kania, M.J.; Homan, F.J.; Mehner, A.W.


    Two methods for measuring failure fraction on irradiated coated-particle fuels have been developed, one in the United States (the IMGA system - Irradiated-Microsphere Gamma Analyzer) and one in the Federal Republic of Germany (FRG) (the PIAA procedure - Postirradiation Annealing and Beta Autoradiography). A comparison of the two methods on two standardized sets of irradiated particles was undertaken to evaluate the accuracy, operational procedures, and expense of each method in obtaining statistically significant results. From the comparison, the postirradiation examination method employing the IMGA system was found to be superior to the PIAA procedure for measuring statistically significant failure fractions. Both methods require that the irradiated fuel be in the form of loose particles, each requires extensive remote hot-cell facilities, and each is capable of physically separating failed particles from unfailed particles. Important differences noted in the comparison are described.

  10. Evaluation of [{sup 11}C]RTI-121 as a selective radioligand for PET studies of the dopamine transporter

    Energy Technology Data Exchange (ETDEWEB)

    Hume, Susan P.; Luthra, Sajinder K.; Brown, David J.; Opacka-Juffry, Jolanta; Osman, Safiye; Ashworth, Sharon; Myers, Ralph; Brady, Frank; Carroll, F. Ivy; Kuhar, Michael J.; Brooks, David J


    The cocaine analogue RTI-121 (3{beta}-(4-iodophenyl)tropane-2{beta}-carboxylic acid isopropyl ester), when labeled with carbon-11, was evaluated in rats as a potential PET ligand for the dopamine transporter. The compound gave in vivo striatum:cerebellum ratios that were similar to those obtained with the related ligand [{sup 11}C]RTI-55 (2{r_reversible}-(4-iodophenyl)tropane-2{beta}-carboxylic acid methyl ester) but showed a much greater selectivity for the dopamine compared with the 5-HT uptake site. The results indicate that [{sup 11}C]RTI-121 could be used in preference to [{sup 11}C]RTI-55 in man. Experimentally, [{sup 11}C]RTI-121 has potential in the quantification of dopamine terminal function in rat models of disease, using a combination of autoradiography, postmortem sampling, and in vivo tomography.

  11. Effect of epileptogenic agents on the incorporation of /sup 3/H-glycine into proteins in the cat's cerebral cortex

    Energy Technology Data Exchange (ETDEWEB)

    Rojik, I.; Feher, O.


    Filter paper strips soaked in /sup 3/H-glycine solution were applied to acoustic cortex of cats, anaesthetized with Nembutal and pretreated with epileptogenic agents (Metrazol, G-penicillin, and 3-amino-pyridine) and cycloheximide. The untreated contralateral hemisphere served as control. After 1 h incubation, both cortical samples were excised simultaneously and fixed in Bouin solution for autoradiography. Incorporation was blocked by cycloheximide. There was no glycine incorporation on the penicillin-treated side, while pyramidal cells were intensively labelled in layers II-V of the mirror focus. 3-Aminopyridine produced the same result. Metrazol as convulsant proved to be far weaker than the previous two. The intensity of incorporation was significantly more intensive in the mirror focus than in the primary one. Penicillin and 3-aminopyridine, while provoking cortical seizures, seem to inhibit glycine incorporation into a neuron-specific, function-dependent protein contained by the labelled cells in the autoradiogram.

  12. Differential regulation of alpha7 nicotinic receptor gene (CHRNA7) expression in schizophrenic smokers. (United States)

    Mexal, Sharon; Berger, Ralph; Logel, Judy; Ross, Randal G; Freedman, Robert; Leonard, Sherry


    The alpha7 neuronal nicotinic receptor gene (CHRNA7) has been implicated in the pathophysiology of schizophrenia by genetic and pharmacological studies. Expression of the alpha7* receptor, as measured by [(125)I]alpha-bungarotoxin autoradiography, is decreased in postmortem brain of schizophrenic subjects compared to non-mentally ill controls. Most schizophrenic patients are heavy smokers, with high levels of serum cotinine. Smoking changes the expression of multiple genes and differentially regulates gene expression in schizophrenic hippocampus. We examined the effects of smoking on CHRNA7 expression in the same tissue and find that smoking differentially regulates expression of both mRNA and protein for this gene. CHRNA7 mRNA and protein levels are significantly lower in schizophrenic nonsmokers compared to control nonsmokers and are brought to control levels in schizophrenic smokers. Sufficient protein but low surface expression of the alpha7* receptor, seen in the autoradiographic studies, suggests aberrant assembly or trafficking of the receptor.

  13. Extreme dryness and DNA-protein cross-links (United States)

    Bieger-Dose, A.; Dose, K.; Meffert, R.; Mehler, M.; Risi, S.

    Exposure of fungal conidia (Aspergillus ochraceus) or spores of Bacillus subtilis to extreme dryness or vacuum induces DNA lesions, including strand breaks and the formation of DNA-protein cross-links. In wet cells only a small amount of protein is bound to DNA, but exposure to conditions of lowered water activity results in an increasing number of cross-links between DNA and proteins. In fungal conidia these cross-links are detected after selective iodination (125J) of the DNA-bound proteins followed by gel electrophoresis and subsequent autoradiography. Another approach is the labelling of DNA with 32p by means of nick translation and the detection of differences in the electrophoretic mobility of DNA before and after digestion with proteinase K of proteins bound to DNA.

  14. Nucleic acid and protein synthesis during lateral root initiation in Marsilea quadrifolia (Marsileaceae) (United States)

    Lin, B. L.; Raghavan, V.


    The pattern of DNA, RNA, and protein synthesis during lateral root initiation in Marsilea quadrifolia L. was monitored by autoradiography of incorporated of 3H-thymidine, 3H-uridine, and 3H-leucine, respectively. DNA synthesis was associated with the enlargement of the lateral root initial prior to its division. Consistent with histological studies, derivatives of the lateral root initial as well as the cells of the adjacent inner cortex and pericycle of the parent root also continued to synthesize DNA. RNA and protein synthetic activities were found to be higher in the lateral root initials than in the endodermal initials of the same longitudinal layer. The data suggest a role for nucleic acid and protein synthesis during cytodifferentiation of a potential endodermal cell into a lateral root initial.

  15. Plasmodium falciparum: characterization of toxin-associated proteins and identification of a hemoglobin containing parasite cytokine stimulator

    DEFF Research Database (Denmark)

    Kristensen, G; Jakobsen, P H


    ]-methionine and immunoprecipitated the labeled antigens with an antiserum against IMP which blocks malaria parasite-induced TNF production. We detected four proteins associated with IMP when the immunoprecipitates were separated by SDS-PAGE and analyzed by autoradiography. To evaluate the capacity of different P. falciparum...... antigens to induce cytokine production we separated a mixture of exoantigens by SDS-PAGE gels. Antigen fractions of 43-71 kDa and of a low molecular mass of TNF alpha interleukin 1 alpha, and interleukin 6 production from human mononuclear cells. The low......Previous studies have indicated the inositol monophosphate (IMP) is a component of the malaria parasite toxin that induces cytokines such as tumour necrosis factor (TNF). To further characterize the toxin we have labeled Plasmodium falciparum in vitro cultures with [14C]inositol or [35S...

  16. Early tissue distribution of therapeutic 131I in a patient with thyrotoxicosis. (United States)

    Vachal, E; Wegst, A; Preston, D; Tomita, T


    A therapeutic dose of 131I was given to a 53-year-old man with thyrotoxicosis three and one-half hours prior to death. The distribution of 131I in the thyroid gland was studied by autoradiography, and the radioactivity in various fresh tissues was measured. Radioactivity was distributed evenly throughout the colloid; small amounts were present in follicular cells. Radioactivity was not localized to the periphery of colloid follicles as observed in an earlier report. Papillary projections were associated with increased radioactivity. Decreased radioactivity was associated with oxyphilic cells. Measurement of 131I in individual tissues showed that per gram of tissue the thyroid gland contained approximately 50 times that of intestine and spleen and approximately 100 times that of liver and bone.

  17. A New Positron Emission Tomography (PET) Radioligand for Imaging Sigma-1 Receptors in Living Subjects

    DEFF Research Database (Denmark)

    James, Michelle L; Shen, Bin; Zavaleta, Cristina L;


    Sigma-1 receptor (S1R) radioligands have the potential to detect and monitor various neurological diseases. Herein we report the synthesis, radiofluorination and evaluation of a new S1R ligand 6-(3-fluoropropyl)-3-(2-(azepan-1-yl)ethyl)benzo[d]thiazol-2(3H)-one ([(18)F]FTC-146, [(18)F]13). [(18)F......]13 was synthesized by nucleophilic fluorination, affording a product with >99% radiochemical purity (RCP) and specific activity (SA) of 2.6 ± 1.2 Ci/µmol (n = 13) at end of synthesis (EOS). Positron emission tomography (PET) and ex vivo autoradiography studies of [(18)F]13 in mice showed high uptake...

  18. Batteries and fuel cells: Design, employment, chemistry (United States)

    Euler, K.-J.

    The history of electrochemical current sources is considered along with primary cells, standard cells, high-energy primary cells, high-energy storage batteries, and fuel cells. Aspects of battery research and development are also discussed, taking into account general considerations related to technological development projects, the introduction of mathematical methods into battery research, resistance measurements, autoradiography and other radiochemical methods, color photography as an aid in research, electron microscopy, X-ray and electron diffraction, spin resonance methods, and electrical measurements involving powders. Attention is given to zinc/manganese dioxide cells, zinc/mercury cells, zinc/silver oxide primary cells, cells utilizing atmospheric oxygen, lead-acid batteries, nickel-iron and nickel-cadmium storage batteries, zinc/silver storage batteries, dry cells with organic depolarizers, dry cells with solid electrolyte, and storage batteries utilizing hydrogen.

  19. Changes in 5-HT4 receptor and 5-HT transporter binding in olfactory bulbectomized and glucocorticoid receptor heterozygous mice

    DEFF Research Database (Denmark)

    Licht, Cecilie Löe; Kirkegaard, Lisbeth; Zueger, Maha;


    The 5-HT(4) receptor is a new potential target for antidepressant treatment and may be implicated in the pathogenesis of depression. This study investigated differences in 5-HT(4) receptor and 5-HT transporter (5-HTT) binding by quantitative autoradiography of [(3)H]SB207145 and (S)-[N-methyl-(3)H......]citalopram in two murine models of depression-related states, olfactory bulbectomy and glucocorticoid receptor heterozygous (GR(+/-)) mice. The olfactory bulbectomy model is characterized by 5-HT system changes, while the GR(+/-) mice have a deficit in hypothalamic-pituitary-adrenal (HPA) system control....... The olfactory bulbectomized mice displayed increased activity in the open field test, a characteristic depression-like feature of this model. After bulbectomy, 5-HT(4) receptor binding was increased in the ventral hippocampus (12%) but unchanged in the dorsal hippocampus, frontal and caudal caudate putamen...

  20. Evaluation of the novel 5-HT4 receptor PET ligand [11C]SB207145 in the Göttingen minipig

    DEFF Research Database (Denmark)

    Kornum, Birgitte R; Lind, Nanna M; Gillings, Nic;


    This study investigates 5-hydroxytryptamine 4 (5-HT(4)) receptor binding in the minipig brain with positron emission tomography (PET), tissue homogenate-binding assays, and autoradiography in vitro. The cerebral uptake and binding of the novel 5-HT(4) receptor radioligand [(11)C]SB207145 in vivo...... autoradiographic 5-HT(4) receptor distribution resembles the human 5-HT(4) receptor distribution with the highest binding in the striatum and no detectable binding in the cerebellum. We found that in the minipig brain [(11)C]SB207145 follows one-tissue compartment kinetics, and the simplified reference tissue...... model provides stable and precise estimates of the binding potential in all regions. The binding potentials calculated for striatum, midbrain, and cortex from the PET data were highly correlated with 5-HT(4) receptor concentrations determined in brain homogenates from the same regions, except...

  1. The brain 5-HT4 receptor binding is down-regulated in the Flinders Sensitive Line depression model and in response to paroxetine administration

    DEFF Research Database (Denmark)

    Licht, Cecilie Löe; Marcussen, Anders Bue; Wegener, Gregers


    The 5-hydroxytryptamine (5-HT(4)) receptor may be implicated in depression and is a new potential target for antidepressant treatment. We have investigated the brain 5-HT(4) receptor [(3)H]SB207145 binding in the Flinders Sensitive Line rat depression model by quantitative receptor autoradiography....... In the Flinders Sensitive Line, the 5-HT(4) receptor and 5-HT transporter binding were decreased in the dorsal and ventral hippocampus, and the changes in binding were directly correlated within the dorsal hippocampus. Chronic but not acute paroxetine administration caused a 16-47% down-regulation of 5-HT(4...... cortices after chronic paroxetine administration, and markedly reduced in several regions after 5-HT depletion. Thus, the 5-HT(4) receptor binding was decreased in the Flinders Sensitive Line depression model and in response to chronic paroxetine administration....

  2. Distribution of /sup 14/C-ochratoxin A in the rainbow trout (Salmo gairdneri)

    Energy Technology Data Exchange (ETDEWEB)

    Fuchs, R.; Appelgren, L.-E.; Hult, K.


    The nephrotoxic mycotoxin ochratoxin A was studied in rainbow trout by whole-body autoradiography and scintillation counting using /sup 14/C-labelled toxin. After one single intravenous injection of 10 fish, corresponding to 160 ng toxin/g body weight, the tissue affinity was studied during an eight day period. As soon as 5 min. after injection the concentration of the radioactivity in the blood had dropped to one tenth of that in the kidney and the urinary bladder. The autoradiograms showed two patterns of blackening in the kidney, one diffuse in the pronephros and one very strong blackening in the opistonephros. In addition to the kidney very high concentrations of radioactivity were also noticed in the bile and the pseudobranch. The muscular tissue of treated trouts contained almost no radioactivity during the whole experiment. Chemical analysis revealed that the radioactivity that could be extracted from the organs was mainly ochratoxin A.

  3. Significance of adrenergic receptors for the development of nevus flammeus and nevus anemicus

    Energy Technology Data Exchange (ETDEWEB)

    Raff, M. (Vienna Univ. (Austria). 2. Hautklinik)


    Examination of patients with nevus flammeus or nevus anemicus showed disturbed sensibility in the area of the nevus in the majority of cases. Histologically and with special technique of histochemistry and fluorescence microscopy there was no evidence for neurogenic lesions. However, signs of vegetative disfunction were present: hyperhidrosis and absent reactivity of vasculature in the nevus area to vasoconstrictive and vasodilatatory stimuli. Based on these findings a disturbed regulation of vascular intramural adrenergic receptors seemed possible and really could be demonstrated by means of autoradiography. In both types of nevi only one of the adrenergic receptors could be marked with specific antagonists. Therefore, the persistent vascular dilatation and constriction can be accounted for by the absence of one of these receptors. This abnormal distribution of receptors could be due to a developmental defect influenced by the ''nerve growth factor''.

  4. Kinetics of circulating B lymphocytes in human myeloma

    Energy Technology Data Exchange (ETDEWEB)

    Boccadoro, M.; Gavarotti, P.; Fossati, G.; Massaia, M.; Pileri, A.; Durie, B.G.


    The tritiated thymidine labeling index (LI%) of peripheral B lymphocytes was studied in eight myeloma patients using simultaneous immunofluorescence and autoradiography. The LI% values were low (0.3%-5.1%), but significantly increased as compared to normal controls. In addition, there was excellent correlation between the LI% values and myeloma disease activity: lowest LI% values were observed in remission patients and the highest at the time of relapse. Simultaneous LI% evaluation of bone marrow myeloma cells in five patients gave concordant results, indicating the same kinetic behavior in both these compartments, particularly in the relapse phase. These data indicate both that circulating B lymphocytes include the neoplastic clone and that these B lymphocytes and bone marrow myeloma cells have similar kinetics.

  5. Nucleolar re-activation is delayed in mouse embryos cloned from two different cell lines

    DEFF Research Database (Denmark)

    Svarcova, Olga; Dinnyes, A.; Polgar, Z.


    displayed early NPBs transformation. In conclusion, despite normal onset of EGA in cloned embryos, activation of functional nucleoli was one cell cycle delayed in NT embryos. NT-MEF embryos displayed normal targeting but delayed activation of nucleolar proteins. Contrary, in NT-HM1 embryos, both......Aim of this study was to evaluate and compare embryonic genome activation (EGA) in mouse embryos of different origin using nucleolus as a marker. Early and late 2-cell and late 4-cell stage embryos, prepared by in vitro fertilization (IVF), parthenogenetic activation (PG), and nuclear transfer...... ofmouse embryonic fibroblast (MEF) and mouse HM1 emryonic stem cells (HM1), were processed for autoradiography following 3H-uridine incubation (transcriptional activity), transmission electron microscopy (ultrastructure) and immunofluorescence (nucleolar proteins; upstream binding factor, UBF...

  6. Subgrains and boron distribution of low carbon bainitic steels

    Institute of Scientific and Technical Information of China (English)

    Xuemin Wang; Bing Cao; Chengjia Shang; Xueyi Liu; Xinlai He


    The structure variation of deformed austenite during the relaxation stage after deformation at various temperatures in an Nb-B ultra low carbon bainitic steel and Fe-Ni alloy was studied by the thermo-simulation. Optical microscope and TEM were applied to analyze the microstructure after RPC (Relaxation-precipitation-controlling phase transformation technique) and the evolution of dislocation configuration. The particle tracking autoradiography (PTA) technique, revealing the distribution of boron, was employed to show the change of boron segregation after different relaxation times. The results indicate that during the relaxation stage the recovery occurs in the deformed austenite, the dislocations rearrange and subgrains form. During the subsequent cooling the boron will segregate at the boundaries of subgrains.

  7. Biokinetic models for radionuclides in experimental animals; Modelos biocineticos de radionucleidos en animales de experimentacion

    Energy Technology Data Exchange (ETDEWEB)

    Morcillo, M. A. [Ciemat. Madrid (Spain)


    The biokinetic models for many radionuclides are, to a large extent, based on data obtained in experimental animals. The methods used in the experimental development of a biokinetic model can be classified in two groups (i) those applied during the experimental work, which include the activity determination of a given radionuclide at different times and in different biological media such as blood, serum, organs/tissues, urine, bile and faeces and (ii) those methods used for the analysis and study of the experimental data, based in mathematical tools. Some of these methods are reviewed,with special emphasis in the whole body macro autoradiography. To conclude, the contribution that this type of studies can have in two fields of radiation protection is discussed, namely optimization of dosimetric evaluations and decorporation of radionuclides. (Author)

  8. A micro-PET/CT approach using O-(2-[{sup 18}F]fluoroethyl)-L-tyrosine in an experimental animal model of F98 glioma for BNCT

    Energy Technology Data Exchange (ETDEWEB)

    Menichetti, L., E-mail: [CNR Institute of Clinical Physiology, Pisa (Italy); Petroni, D.; Panetta, D. [CNR Institute of Clinical Physiology, Pisa (Italy); Burchielli, S. [Fondazione CNR/Regione Toscana G. Monasterio, Pisa (Italy); Bortolussi, Silva [Dept. Theoretical and Nuclear Physics, University of Pavia, Pavia (Italy); Matteucci, M. [Scuola Superiore Sant' Anna, Pisa (Italy); Pascali, G.; Del Turco, S. [CNR Institute of Clinical Physiology, Pisa (Italy); Del Guerra, A. [Department of Physics, University of Pisa, Pisa (Italy); Altieri, S. [Dept. Theoretical and Nuclear Physics, University of Pavia, Pavia (Italy); Salvadori, P.A. [CNR Institute of Clinical Physiology, Pisa (Italy)


    The present study focuses on a micro-PET/CT application to be used for experimental Boron Neutron Capture Therapy (BNCT), which integrates, in the same frame, micro-CT derived anatomy and PET radiotracer distribution. Preliminary results have demonstrated that {sup 18}F-fluoroethyl-tyrosine (FET)/PET allows the identification of the extent of cerebral lesions in F98 tumor bearing rat. Neutron autoradiography and {alpha}-spectrometry on axial tissues slices confirmed the tumor localization and extraction, after the administration of fructose-boronophenylalanine (BPA). Therefore, FET-PET approach can be used to assess the transport, the net influx, and the accumulation of FET, as an aromatic amino acid analog of BPA, in experimental animal model. Coregistered micro-CT images allowed the accurate morphological localization of the radiotracer distribution and its potential use for experimental BNCT.

  9. Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA

    DEFF Research Database (Denmark)

    Klein, A B; Bay, T; Villumsen, I S


    analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations......, (3)H-HOCPCA displays excellent signal-to-noise ratios using rodent brain autoradiography, which makes it a valuable ligand for anatomical quantification of native GHB binding site levels. Our data confirmed that (3)H-HOCPCA labels only the high-affinity specific GHB binding site, found in high...... density in cortical and hippocampal regions. The experiments revealed markedly stronger binding at pH 6.0 (Kd 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar Bmax values. Using (3)H-HOCPCA we analyzed the GHB binding protein profile during mouse...

  10. Distribution of i.v. administered epidermal growth factor in the rat

    DEFF Research Database (Denmark)

    Poulsen, Steen Seier; Nexø, Ebba


    The distribution of i.v. injected 125I-labeled epidermal growth factor (EGF) was examined in the rat. The uptake of radioactivity was examined for the following tissues: liver, kidney, skin, stomach, small intestine, colon, brain, submandibular gland, lung, spleen, and testis. 125I-EGF was cleared...... of 125I-EGF per g tissue was markedly higher for the liver and kidneys than for the rest of the organs. By autoradiography 125I-EGF was found in the peripheral parts of the classical liver lobule, in the proximal tubules of the kidneys, in the surface epithelium of the stomach, and in the surface...... epithelium of the villi in the small intestine. In conclusion the present study showed that small doses of homologous EGF was cleared from the circulation of rats within minutes, mainly by the liver, the kidneys, and the small intestine....

  11. Mapping CB1 cannabinoid receptors with [3H]OMAR in the Flinders rodent model of depression

    DEFF Research Database (Denmark)

    Nahimi, A.; Gjedde, A.; Wong, D. F.


    ). In these rats, CB1 receptor autoradiography in postmortem tissue was carried out by incubating 20 mM thick sagital sections in the presence of the CB1 receptor antagonist [3H] OMAR (method adapted from Mato & Pazos 2004, with small modifications). Non-specific binding was determined on adjacent sections...... in the presence of [3H]OMAR and the CB receptor agonist CP55,940. Results: Eighteen regions of interest were analyzed on the autoradiogram. The highest binding (in units of pmol/g(plus or minus)SD) was found in the hippocampus (105(plus or minus)21 in FSL and 105(plus or minus)16 in FRL), striatum (69(plus...

  12. Metabolic changes in deafferented central neurons of an insect, Acheta domesticus. I. Effects upon amino acid uptake and incorporation

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, M.R.; Edwards, J.S.


    Chronic cercal deafferentation of the terminal ganglion in developing crickets (Acheta domesticus), which is known to suppress normal development of giant interneuron dendritic arborizations is shown here to reduce (/sup 3/H)leucine uptake and incorporation into ganglion proteins. Short term deafferentation of adult crickets, in contrast, does not depress amino acid uptake and incorporation significantly. Following unilateral long term deafferentation of the terminal ganglion, a comparison was made of the (/sup 3/H)leucine incorporation into primary dendritic processes and somata of deafferented and normally innervated medial giant interneurons (MGIs) within the same ganglion by means of quantitative autoradiography. Grain densities within dendrites of deafferented MGIs were significantly lower than in paired control MGIs' grain densities within somata of deafferented MGIs also were reduced, although the effects of deafferentation were less pronounced in somata than in target dendrites. These results imply a specific influence of afferent innervation on protein metabolism during growth and development of target postsynaptic elements.

  13. Effect of Dachengqi Tang(大承气汤)Granule on Proliferation of Intestinal Epithelial Cells in Rats with Experimental Intestinal Obstruction

    Institute of Scientific and Technical Information of China (English)

    KANGYi; LINXiu-zhen


    Objective:To study the effects of Dachengqi Tang(DCQT) granule on the proliferation of the intestinal epithelial cells in rats with experimental intestinal obstruction.Methods:Experimental intes-tinal obstruction models were established in rats and autoradiography with 3H-TdR was used to determine 3H-TdR labeling counts of intestinal epithelial cells in rats.Results:DCQT granule had no effects on 3H-TdR labeling counts of intestinal epithelial cells in normal rats.DCQT granule obviously increases the rate of renovation in intestinal epithelial cells of the intestinal obstruction rats.Conclusion:DCQT granule could reinforce the intestinal mucosa's defensive function by means of increasing the proliferation of intesti-nal epithelial cells.

  14. Effect of Dachengqi Tang (大承气汤) Granule on Proliferation of Intestinal Epithelial Cells in Rats with Experimental Intestinal Obstruction

    Institute of Scientific and Technical Information of China (English)

    康毅; 林秀珍


    Objective: To study the effects of Dachengqi Tang (DCQT) granule on the proliferation of the intestinal epithelial cells in rats with experimental intestinal obstruction. Methods: Experimental intestinal obstruction models were established in rats and autoradiography with 3H-TdR was used to determine 3H-TdR labeling counts of intestinal epithelial cells in rats. Results: DCQT granule had no effects on 3H-TdR labeling counts of intestinal epithelial cells in normal rats. DCQT granule obviously increases the rate of renovation in intestinal epithelial cells of the intestinal obstruction rats. Conclusion: DCQT granule could reinforce the intestinal mucosa's defensive function by means of increasing the proliferation of intestinal epithelial cells.

  15. Possible recruitment of osteoblastic precursor cells from hypertrophic chondrocytes during initial osteogenesis in cartilaginous limbs of young rats. (United States)

    Franzen, A; Oldberg, A; Solursh, M


    The appearance of the bone phenotype during rat embryogenesis was studied by in situ hybridization using a cDNA clone to osteopontin. Radiolabeled sense and antisense RNA probes were prepared from the osteopontin cDNA by in vitro transcription. The probes were used to hybridize paraffin sections of the cartilaginous diaphysis from embryonic rats at day 17 of gestation. The hybridization pattern was analyzed by autoradiography. Hybridization with the antisense probe gave patterns of silver grain labeling, indicating the presence of osteopontin mRNA among the hypertrophic chondrocytes. No silver grains could be detected in the corresponding region following hybridization of consecutive sections with the sense probe, showing the specificity of the technique being used. Whether these results indicate that the osteopontin gene is transiently expressed by hypertrophic chondrocytes or that osteopontin is an early marker for osteoblastic precursor cells will have to be explored further.

  16. Synthesis and evaluation of boron compounds for neutron capture therapy of malignant brain tumors. Technical progress report No. 1, May 1, 1990--January 31, 1991

    Energy Technology Data Exchange (ETDEWEB)

    Soloway, A.H.; Barth, R.F.


    Boron neutron capture therapy offers the potentiality for treating brain tumors currently resistant to treatment. The success of this form of therapy is directly dependent upon the delivery of sufficient numbers of thermal-neutrons to tumor cells which possess high concentrations of B-10. The objective of this project is to develop chemical methodology to synthesize boron-containing compounds with the potential for becoming incorporated into rapidly-dividing malignant brain tumor cells and excluded from normal components of the brain and surrounding tissues, to develope biological methods for assessing the potential of the compound by use of cell culture or intratumoral injection, to develop analytical methodology for measuring boron in cells and tissue using direct current plasma atomic emission spectroscopy (DCP-AES) and alpha track autoradiography, to develop biochemical and HPLC procedures for evaluating compound uptake and tissue half-life, and to develop procedures required to assess both in vitro and vivo efficacy of BNCT with selected compounds.

  17. Synthesis and evaluation of boron compounds for neutron capture therapy of malignant brain tumors

    Energy Technology Data Exchange (ETDEWEB)

    Soloway, A.H.; Barth, R.F.


    Boron neutron capture therapy offers the potentiality for treating brain tumors currently resistant to treatment. The success of this form of therapy is directly dependent upon the delivery of sufficient numbers of thermal-neutrons to tumor cells which possess high concentrations of B-10. The objective of this project is to develop chemical methodology to synthesize boron-containing compounds with the potential for becoming incorporated into rapidly-dividing malignant brain tumor cells and excluded from normal components of the brain and surrounding tissues, to develope biological methods for assessing the potential of the compound by use of cell culture or intratumoral injection, to develop analytical methodology for measuring boron in cells and tissue using direct current plasma atomic emission spectroscopy (DCP-AES) and alpha track autoradiography, to develop biochemical and HPLC procedures for evaluating compound uptake and tissue half-life, and to develop procedures required to assess both in vitro and vivo efficacy of BNCT with selected compounds.

  18. Bioconcentration of the anionic surfactant linear alkylbenzene sulfonate (LAS) in the marine shrimp Palaemonetes varians: a radiotracer study. (United States)

    Renaud, Florent; Warnau, Michel; Oberhänsli, François; Teyssié, Jean-Louis; Temara, Ali; Rouleau, Claude; Metian, Marc


    Uptake and depuration kinetics of dissolved [(14)C]C₁₂-6-linear alkylbenzene sulfonate (LAS) were determined in the shrimp Palaemonetes varians using environmentally relevant exposure concentration. The shrimp concentrated LAS from seawater with a mean BCF value of 120 L kg(-1) after a 7-day exposure. Uptake biokinetics were best described by a saturation model, with an estimated BCFss, of 159 ± 34 L kg(-1), reached after 11.5 days. Shrimp weight influenced significantly BCF value with smaller individuals presenting higher affinity to LAS. To the light of a whole body autoradiography, major accumulation of LAS occurred in the cephalothorax circulatory system (gills, heart, hepatopancreas) and ocular peduncle, but not in the flesh, limiting potential transfer to human consumers. LAS depuration rate constant value of the shrimp was 1.18 ± 0.08 d(-1) leading to less than 1% of remaining LAS in its tissues after 8 days of depuration.

  19. [{sup 125}I]{beta}-CIT-FE and [{sup 125}I]{beta}-CIT-FP are superior to [{sup 125}I]{beta}-CIT for dopamine transporter visualization: Autoradiographic evaluation in the human brain

    Energy Technology Data Exchange (ETDEWEB)

    Guenther, Ilonka; Hall, Haakan; Halldin, Christer; Swahn, Carl-Gunnar; Farde, Lars; Sedvall, Goeran


    The binding of the three dopamine transporter radioligands ([{sup 125}I]{beta}-CIT, [{sup 125}I]{beta}-CIT-FE, and [{sup 125}I]{beta}-CIT-FP) was studied using whole-hemisphere autoradiography on postmortem human brains. The autoradiograms revealed an intense and homogeneous labeling of the nucleus caudatus and putamen but also to varying extent to serotonergic and noradrenergic transporters of neocortex and thalamus. The order of specificity estimated (striatum over neocortex ratios) was {beta}-CIT-FP > {beta}-CIT-FE >> {beta}-CIT, suggesting that {beta}-CIT-FE and {beta}-CIT-FP should be preferred for in vivo studies of the dopamine transporter in the human brain.

  20. Effects of carbon dioxide on Penicillium chrysogenum: an autoradiographic study

    Energy Technology Data Exchange (ETDEWEB)

    Edwards, A.G.; Ho, C.S.


    Previous research has shown that dissolved carbon dioxide causes significant changes in submerged penicillin fermentations, such as stunted, swollen hyphae, increased branching, lower growth rates, and lower penicillin productivity. Influent carbon dioxide levels of 5 and 10% were shown through the use of autoradiography to cause an increase in chitin synthesis in submerged cultures of Penicillium chrysogenum. At an influent 5% carbon dioxide level, chitin synthesis is ca. 100% greater in the subapical region of P. chrysogenum hyphae than that of the control, in which there was no influent carbon dioxide. Influent carbon dioxide of 10% caused an increase of 200% in chitin synthesis. It is believed that the cell wall must be plasticized before branching can occur and that high amounts of dissolved carbon dioxide cause the cell to lose control of the plasticizing effect, thus the severe morphological changes occur.

  1. Radioiodinated benzodiazepines: agents for mapping glial tumors

    Energy Technology Data Exchange (ETDEWEB)

    Van Dort, M.E.; Ciliax, B.J.; Gildersleeve, D.L.; Sherman, P.S.; Rosenspire, K.C.; Young, A.B.; Junck, L.; Wieland, D.M.


    Two isomeric iodinated analogues of the peripheral benzodiazepine binding site (PBS) ligand Ro5-4864 have been synthesized and labeled in high specific activity with iodine-125. Competitive binding assays conducted with the unlabeled analogues indicate high affinity for PBS. Tissue biodistribution studies in rats with these /sup 125/I-labeled ligands indicate high uptake of radioactivity in the adrenals, heart, and kidney--tissues known to have high concentrations of PBS. Preadministration of the potent PBS antagonist PK 11195 blocked in vivo uptake in adrenal tissue by over 75%, but to a lesser degree in other normal tissues. In vivo binding autoradiography in brain conducted in C6 glioma bearing rats showed dense, PBS-mediated accumulation of radioactivity in the tumor. Ligand 6 labeled with /sup 123/I may have potential for scintigraphic localization of intracranial glioma.

  2. Cytophysiological and ultrastructural modifications induced by cold in the microsporocytes and tapetum of Rhoeo discolor Hance

    Directory of Open Access Journals (Sweden)

    André Souvré


    Full Text Available The exposition of Rhoeo discolor to cold induces an alteration of the microsporocytes (PMC and tapetum ultrastructure. In the young cooled PMC, the mitochondria present short and vesiculate cristae, the stroma of proplasts is clearer and the polyrilbasomes are deteriorated. During the phase tetrads-microspores, the alterations are more important: the chromatin coagulates, the nucleus swells while the nuclear membrane is modified; some large vesicules appear outside of the plasmalemma. In the cooled periplasmodium we can observe many groups of vesicules, mitochondria with dilated cristae, rough endoplasmic reticulum without their ribosomes and a breaking up of inueleoli. Our observations are in correlation with the previous results obtained by autoradiography and photometry, and are discussed with the bibliographical results.

  3. Interaction of minoxidil with pigment in cells of the hair follicle: an example of binding without apparent biological effects. (United States)

    Buhl, A E; Kawabe, T T; MacCallum, D K; Waldon, D J; Knight, K A; Johnson, G A


    To identify minoxidil target cells in hair follicles we followed the uptake of radiolabeled drug in mouse vibrissae follicles both in vitro and in vivo. Autoradiography showed that both 3H-minoxidil and 3H-minoxidil sulfate accumulated in the differentiating epithelial matrix cells superior to the dermal papilla, a distribution similar to that of pigment. Minoxidil localized in melanocytes, melanocyte processes, and areas of greater melanin concentrations within the epithelial cells. Although uptake of minoxidil was significantly less in unpigmented follicles, the drug stimulated proliferation and differentiation of both pigmented and unpigmented follicles. Labeled minoxidil bound to Sepia melanin and was displaced with unlabeled minoxidil and other electron donor drugs. This interaction with melanin acts as a targeting mechanism of minoxidil to pigmented hair follicles but has no apparent functional significance in hair growth. This work illustrates how measurement of drugs in hair may be biased by pigmentation.

  4. Simple detection of hepatitis C virus using {sup 125}I-2'-deoxyuridine triphosphate and gamma counter

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Soo Jin; Ahn, S. H.; Chung, W. S.; Woo, K. S.; Lim, S. J.; Choi, C. W.; Lim, S. M. [Korea Cancer Center Hospital, Seoul (Korea, Republic of)


    Hepatitis C Virus (HCV) is the major cause of post transfusion and sporadic non A, non B hepatitis. Current infection of HCV can be detected by PCR method. Using PCR, it has been possible to detect HCV viremia prior to immunological sero-conversion and to detect fluctuation of viremia in antibody-positive chronic HCV patients undergoing therapy with interferon. In this study, we established the simple method to detect HCV DNA by incorporation of {sup 125}I-deoxyuridine triphosphate(dUTP) into DNA during the PCR, and counted the radioactivity of PCR product by gamma counter. {sup 125}I-2'-deoxyuridine 5'-triphosphate was prepared, and incorporated into DNA during PCR. dUTP was radiolabeled by the iododemercuration of 5-mercuri intermediate. Iododemercuration labeling was completed with 98% yield and the obtained product was incorporated into DNA without further purification. After incorporation, covalently bonded radioiodine substituent was remained stable during PCR procedure HCV positive standard and positive patient sera in immunological assay were centrifuged. HCV RNA is isolated from by GTC(Guanidine Thiocyanate) and phenol/chloroform extraction method and synthesized complementary DNA by using reverse transcriptase. The '1{sup 25}I-dUTP was incorporated into HCV C DNA during PCR. PCR product purified by fiber matrix column and counted by gamma counter. PCR products were electrophoresized, and autoradiography image obtained. Amplified HCV DNA by {sup 125}I-dUTP PCR obtained the band on the gel by electrophoresis and autoradiography at the same position. In patient sera, radioactivity of HCV positive sample was 8 times higher than HCV negative viremia sample. We established HCV detection method using {sup 125}I-dUTP. {sup 125}I-dUTP PCR detection of HCV is convenient and reporducible.

  5. Pituitary binding and internalization of radioiodinated gonadotropin-releasing hormone agonist and antagonist ligands in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Wynn, P.C.; Suarez-Quian, C.A.; Childs, G.V.; Catt, K.J.


    In rat pituitary gonadotrophs, the rates of binding and endocytosis of two GnRH superagonist analogs, (D-Ala6,Pro9-NEt)GnRH and (D-Lys6,Pro9-NEt)GnRH, were compared with those of the potent antagonist analog (N-acetyl-D-pCl-Phe1,2,D-Trp3,D-Lys6,D-Ala10)GnRH by quantitative electron microscopic autoradiography. In dispersed pituitary cells, the two agonist analogs showed similar binding kinetics and comparable degrees of sequestration, as measured by their resistance to dissociation by low pH buffer. However, quantification of silver grain localization suggested that cellular internalization of the (D-Ala6)GnRH agonist increased more rapidly than that of the (D-Lys6)GnRH analog. These discrepancies, and the finding that a larger amount of the specifically bound /sup 125/I-(D-Ala6)GnRH agonist was removed during glutaraldehyde fixation, indicated that the proportional internalization of this analog was over estimated by quantitative autoradiography owing to loss of cell surface-bound radioligand. We, therefore, employed radioiodinated D-Lys6-substituted analogs to analyze the receptor binding and cellular uptake of GnRH agonist and antagonist derivatives in vivo. After iv injection, a high proportion of the /sup 125/I-(D-Lys6)GnRH agonist was translocated into pituitary gonadotrophs within 60 min, whereas the D-Lys6 antagonist was predominantly associated with the plasma membrane during that time. Four hours after injection of the antagonist, an appreciable proportion of silver grains was associated with intracellular organelles, and this trend increased progressively at later time points. The relatively prolonged cellular processing of the GnRH antagonist is consistent with in vivo binding kinetics, and its slower internalization may reflect the basal rate of GnRH receptor turnover in the cell membrane.

  6. Sympathetic nerve damage and restoration after ischemia-reperfusion injury as assessed by {sup 11}C-hydroxyephedrine

    Energy Technology Data Exchange (ETDEWEB)

    Werner, Rudolf A.; Higuchi, Takahiro [University of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); University of Wuerzburg, Comprehensive Heart Failure Center, Wuerzburg (Germany); Maya, Yoshifumi [University of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); Nihon Medi-Physics Co., Ltd., Research Centre, Chiba (Japan); Rischpler, Christoph [Technische Universitaet Muenchen, Department of Nuclear Medicine, Klinikum rechts der Isar, Muenchen (Germany); Javadi, Mehrbod S. [Johns Hopkins University, Division of Nuclear Medicine, Russell H. Morgan Department of Radiology, Baltimore, MD (United States); Fukushima, Kazuhito [Hyogo College of Medicine, Department of Radiology, Hyogo (Japan); Lapa, Constantin [University of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); Herrmann, Ken [University of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); David Geffen School of Medicine at UCLA, Department of Molecular and Medical Pharmacology, Los Angeles, CA (United States)


    An altered state of the cardiac sympathetic nerves is an important prognostic factor in patients with coronary artery disease. The aim of this study was to investigate regional sympathetic nerve damage and restoration utilizing a rat model of myocardial transient ischemia and a catecholamine analog PET tracer, {sup 11}C-hydroxyephedrine ({sup 11}C-HED). Transient myocardial ischemia was induced by coronary occlusion for 20 min and reperfusion in male Wistar rats. Dual-tracer autoradiography was performed subacutely (7 days) and chronically (2 months) after ischemia, and in control rats without ischemia using {sup 11}C-HED as a marker of sympathetic innervation and {sup 201}TI for perfusion. Additional serial in vivo cardiac {sup 11}C-HED and {sup 18}F-FDG PET scans were performed in the subacute and chronic phases after ischemia. After transient ischemia, the {sup 11}C-HED uptake defect areas in both the subacute and chronic phases were clearly larger than the perfusion defect areas in the midventricular wall. The subacute {sup 11}C-HED uptake defect showed a transmural pattern, whereas uptake recovered in the subepicardial portion in the chronic phase. Tyrosine hydroxylase antibody nerve staining confirmed regional denervation corresponding to areas of decreased {sup 11}C-HED uptake. Serial in vivo PET imaging visualized reductions in the area of the {sup 11}C-HED uptake defects in the chronic phase consistent with autoradiography and histology. Higher susceptibility of sympathetic neurons compared to myocytes was confirmed by a larger {sup 11}C-HED defect with a corresponding histologically identified region of denervation. Furthermore, partial reinnervation was observed in the chronic phase as shown by recovery of subepicardial {sup 11}C-HED uptake. (orig.)

  7. Memory formation, amnesia, improved memory and reversed amnesia: 5-HT role. (United States)

    Perez-Garcia, G; Meneses, A


    Traditionally, the search for memory circuits has been focused on examinations of amnesic and AD patients, cerebral lesions and neuroimaging. A complementary alternative has become the use of autoradiography with radioligands, aiming to identify neurobiological markers associated with memory formation, amnesia states and (more recently) recovery from memory deficits. Indeed, ex vivo autoradiographic studies offer the advantage of detecting functionally active receptors altered by pharmacological tools during memory formation, amnesia states and memory recovery. Moreover, serotonin (5-hydroxytryptamine, 5-HT) systems have become a pharmacological and genetic target in the treatment of memory disorders. Herein evidence from studies involving expression of 5-HT(1A), 5-HT(2A), 5-HT(4), and 5-HT(6) receptors in memory formation, amnesia conditions (e.g., pharmacological models or aging) and recovery of memory is reviewed. Thus, specific 5-HT receptors were expressed in trained animals relative to untrained in brain areas such as cortex, hippocampus and amygdala. However, relative to the control group, rats showing amnesia or recovered memory, showed in the hippocampus, region where explicit memory is formed, a complex pattern of 5-HT receptor expression. An intermediate expression occurred in amygdala, septum and some cortical areas in charge of explicit memory storage. Even in brain areas thought to be in charge of procedural memory such as basal ganglia, animals showing recovered memory displayed an intermediate expression, while amnesic groups, depending on the pharmacological amnesia model, showed up- or down-regulation. In conclusion, evidence indicates that autoradiography, by using specific radioligands, offers excellent opportunities to map dynamic changes in brain areas engaged in these cognitive processes. The 5-HT modulatory role strengthens or suppresses memory is critically depend on the timing of the memory formation.

  8. Uptake of inflammatory cell marker [{sup 11}C]PK11195 into mouse atherosclerotic plaques

    Energy Technology Data Exchange (ETDEWEB)

    Laitinen, Iina; Marjamaeki, Paeivi; Naagren, Kjell; Roivainen, Anne; Knuuti, Juhani [University of Turku, Turku PET Centre, Turku (Finland); Laine, V.J.O. [Turku University Hospital, Department of Pathology, Turku (Finland); Wilson, Ian [GE Healthcare Biosciences, Medical Diagnostics, London (United Kingdom); Leppaenen, Pia; Ylae-Herttuala, Seppo [University of Kuopio, A.I. Virtanen Institute, Kuopio (Finland)


    The ligand [{sup 11}C]PK11195 binds with high affinity and selectivity to peripheral benzodiazepine receptor, expressed in high amounts in macrophages. In humans, [{sup 11}C]PK11195 has been used successfully for the in vivo imaging of inflammatory processes of brain tissue. The purpose of this study was to explore the feasibility of [{sup 11}C]PK11195 in imaging inflammation in the atherosclerotic plaques. The presence of PK11195 binding sites in the atherosclerotic plaques was verified by examining the in vitro binding of [{sup 3}H]PK11195 onto mouse aortic sections. Uptake of intravenously administered [{sup 11}C]PK11195 was studied ex vivo in excised tissue samples and aortic sections of a LDLR/ApoB48 atherosclerotic mice. Accumulation of the tracer was compared between the atherosclerotic plaques and non-atherosclerotic arterial sites by autoradiography and histological analyses. The [{sup 3}H]PK11195 was found to bind to both the atherosclerotic plaques and the healthy wall. The autoradiography analysis revealed that the uptake of [{sup 11}C]PK11195 to inflamed regions in plaques was more prominent (p = 0.011) than to non-inflamed plaque regions, but overall it was not higher than the uptake to the healthy vessel wall. Also, the accumulation of {sup 11}C radioactivity into the aorta of the atherosclerotic mice was not increased compared to the healthy control mice. Our results indicate that the uptake of [{sup 11}C]PK11195 is higher in inflamed atherosclerotic plaques containing a large number of inflammatory cells than in the non-inflamed plaques. However, the tracer uptake to other structures of the artery wall was also prominent and may limit the use of [{sup 11}C]PK11195 in clinical imaging of atherosclerotic plaques. (orig.)

  9. High-performance imaging of stem cells using single-photon emissions (United States)

    Wagenaar, Douglas J.; Moats, Rex A.; Hartsough, Neal E.; Meier, Dirk; Hugg, James W.; Yang, Tang; Gazit, Dan; Pelled, Gadi; Patt, Bradley E.


    Radiolabeled cells have been imaged for decades in the field of autoradiography. Recent advances in detector and microelectronics technologies have enabled the new field of "digital autoradiography" which remains limited to ex vivo specimens of thin tissue slices. The 3D field-of-view (FOV) of single cell imaging can be extended to millimeters if the low energy (10-30 keV) photon emissions of radionuclides are used for single-photon nuclear imaging. This new microscope uses a coded aperture foil made of highly attenuating elements such as gold or platinum to form the image as a kind of "lens". The detectors used for single-photon emission microscopy are typically silicon detectors with a pixel pitch less than 60 μm. The goal of this work is to image radiolabeled mesenchymal stem cells in vivo in an animal model of tendon repair processes. Single-photon nuclear imaging is an attractive modality for translational medicine since the labeled cells can be imaged simultaneously with the reparative processes by using the dual-isotope imaging technique. The details our microscope's two-layer gold aperture and the operation of the energy-dispersive, pixellated silicon detector are presented along with the first demonstration of energy discrimination with a 57Co source. Cell labeling techniques have been augmented by genetic engineering with the sodium-iodide symporter, a type of reporter gene imaging method that enables in vivo uptake of free 99mTc or an iodine isotope at a time point days or weeks after the insertion of the genetically modified stem cells into the animal model. This microscopy work in animal research may expand to the imaging of reporter-enabled stem cells simultaneously with the expected biological repair process in human clinical trials of stem cell therapies.

  10. Gene activity during germination of spores of the fern, Onoclea sensibilis: RNA and protein synthesis and the role of stored mRNA (United States)

    Raghavan, V.


    Pattern of 3H-uridine incorporation into RNA of spores of Onoclea sensibilis imbibed in complete darkness (non-germinating conditions) and induced to germinate in red light was followed by oligo-dT cellulose chromatography, gel electrophoresis coupled with fluorography and autoradiography. In dark-imbibed spores, RNA synthesis was initiated about 24 h after sowing, with most of the label accumulating in the high mol. wt. poly(A) -RNA fraction. There was no incorporation of the label into poly(A) +RNA until 48 h after sowing. In contrast, photo-induced spores began to synthesize all fractions of RNA within 12 h after sowing and by 24 h, incorporation of 3H-uridine into RNA of irradiated spores was nearly 70-fold higher than that into dark-imbibed spores. Protein synthesis, as monitored by 3H-arginine incorporation into the acid-insoluble fraction and by autoradiography, was initiated in spores within 1-2 h after sowing under both conditions. Autoradiographic experiments also showed that onset of protein synthesis in the cytoplasm of the germinating spore is independent of the transport of newly synthesized nuclear RNA. One-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis of 35S-methionine-labelled proteins revealed a good correspondence between proteins synthesized in a cell-free translation system directed by poly(A) +RNA of dormant spores and those synthesized in vivo by dark-imbibed and photo-induced spores. These results indicate that stored mRNAs of O. sensibilis spores are functionally competent and provide templates for the synthesis of proteins during dark-imbibition and germination.

  11. Evaluation of vegetables in Tsukuba for contamination with radioactive materials from the accident at Fukushima Daiichi nuclear power plant. (United States)

    Isobe, Tomonori; Mori, Yutaro; Takada, Kenta; Sato, Eisuke; Takahashi, Hideki; Sekiguchi, Takao; Yoshimura, Yousuke; Sakurai, Hideyuki; Sakae, Takeji


    A large amount of radioactive material was released into the atmosphere after the accident of the Fukushima Daiichi Nuclear Power Plant following the Tohoku earthquake on 11 March 2011, and traces of these materials were detected in Tsukuba. Because radioactive materials can adhere to vegetables, the authors made a qualitative evaluation of vegetables in Tsukuba, estimated internal exposure dose based on quantitative measurement results, and investigated several decontamination methods. Qualitative analysis of vegetable contamination was done by autoradiography. Quantitative analysis was done using a high-purity germanium detector. To assess decontamination, two methods were tested: one with running water and the other with boiling water. In addition, boiled soup stock was measured. In the qualitative evaluation by autoradiography, radioactive materials were not uniformly distributed but adhered to vegetables in clumps and hot spots. In the quantitative evaluation to measure contamination of outer and inner leaves of sanchu lettuce, it was observed that the concentration of I was 8,031.35 ± 764.79 Bq kg in the outer leaves and 115.28 ± 20.63 Bq kg in the inner leaves. In addition, the concentration of Cs was 1,371.93 ± 366.45 Bq kg in the outer leaves and 9.68 ± 15.03 Bq kg in the inner leaves. This suggests that one can greatly reduce internal exposure dose by removing the outer leaves if one has to eat vegetables just after a nuclear accident. In the decontamination assessment, a decontamination efficiency of up to 70% was achieved by boiling vegetables for 20 min.

  12. Validation of MRI-based 3D digital atlas registration with histological and autoradiographic volumes: an anatomofunctional transgenic mouse brain imaging study. (United States)

    Lebenberg, J; Hérard, A-S; Dubois, A; Dauguet, J; Frouin, V; Dhenain, M; Hantraye, P; Delzescaux, T


    Murine models are commonly used in neuroscience to improve our knowledge of disease processes and to test drug effects. To accurately study neuroanatomy and brain function in small animals, histological staining and ex vivo autoradiography remain the gold standards to date. These analyses are classically performed by manually tracing regions of interest, which is time-consuming. For this reason, only a few 2D tissue sections are usually processed, resulting in a loss of information. We therefore proposed to match a 3D digital atlas with previously 3D-reconstructed post mortem data to automatically evaluate morphology and function in mouse brain structures. We used a freely available MRI-based 3D digital atlas derived from C57Bl/6J mouse brain scans (9.4T). The histological and autoradiographic volumes used were obtained from a preliminary study in APP(SL)/PS1(M146L) transgenic mice, models of Alzheimer's disease, and their control littermates (PS1(M146L)). We first deformed the original 3D MR images to match our experimental volumes. We then applied deformation parameters to warp the 3D digital atlas to match the data to be studied. The reliability of our method was qualitatively and quantitatively assessed by comparing atlas-based and manual segmentations in 3D. Our approach yields faster and more robust results than standard methods in the investigation of post mortem mouse data sets at the level of brain structures. It also constitutes an original method for the validation of an MRI-based atlas using histology and autoradiography as anatomical and functional references, respectively.

  13. Small-Animal PET Imaging of Tau Pathology with 18F-THK5117 in 2 Transgenic Mouse Models. (United States)

    Brendel, Matthias; Jaworska, Anna; Probst, Federico; Overhoff, Felix; Korzhova, Viktoria; Lindner, Simon; Carlsen, Janette; Bartenstein, Peter; Harada, Ryuichi; Kudo, Yukitsuka; Haass, Christian; Van Leuven, Fred; Okamura, Nobuyuki; Herms, Jochen; Rominger, Axel


    Abnormal accumulation of tau aggregates in the brain is one of the hallmarks of Alzheimer disease neuropathology. We visualized tau deposition in vivo with the previously developed 2-arylquinoline derivative (18)F-THK5117 using small-animal PET in conjunction with autoradiography and immunohistochemistry gold standard assessment in 2 transgenic mouse models expressing hyperphosphorylated tau. Small-animal PET recordings were obtained in groups of P301S (n = 11) and biGT mice (n = 16) of different ages, with age-matched wild-type (WT) serving as controls. After intravenous administration of 16 ± 2 MBq of (18)F-THK5117, a dynamic 90-min emission recording was initiated for P301S mice and during 20-50 min after injection for biGT mice, followed by a 15-min transmission scan. After coregistration to the MRI atlas and scaling to the cerebellum, we performed volume-of-interest-based analysis (SUV ratio [SUVR]) and statistical parametric mapping. Small-animal PET results were compared with autoradiography ex vivo and in vitro and further validated with AT8 staining for neurofibrillary tangles. SUVRs calculated from static recordings during the interval of 20-50 min after tracer injection correlated highly with estimates of binding potential based on the entire dynamic emission recordings (R = 0.85). SUVR increases were detected in the brain stem of aged P301S mice (+11%; P parametric mapping analysis. Saturable binding of the tracer was verified by autoradiographic blocking studies. In the first dedicated small-animal PET study in 2 different transgenic tauopathy mouse models using the tau tracer (18)F-THK5117, the temporal and spatial progression could be visualized in good correlation with gold standard assessments of tau accumulation. The serial small-animal PET method could afford the means for preclinical testing of novel therapeutic approaches by accommodating interanimal variability at baseline, while detection thresholds in young animals have to be considered.

  14. {sup 99m}Tc-annexin V and {sup 111}In-antimyosin antibody uptake in experimental myocardial infarction in rats

    Energy Technology Data Exchange (ETDEWEB)

    Sarda-Mantel, Laure; Rouzet, Francois; Martet, Genevieve; Raguin, Olivier; Vrigneaud, Jean-Marc; Guludec, Dominique Le [Bichat Hospital AP-HP, EA 3512, Nuclear Medicine Department, Paris (France); Michel, Jean-Baptiste; Louedec, Liliane [INSERM U460, UFR Bichat, Paris (France); Vanderheyden, Jean-Luc [Theseus Imaging Corporation, Boston, MA (United States); Hervatin, Florence [Bichat Hospital AP-HP, EA 3512, Nuclear Medicine Department, Paris (France); CGA/SHFS, Orsay (France); Khaw, Ban An [Bouve College of Pharmacy and Health Sciences, Center for Drug Targeting and Analysis, Boston, MA (United States)


    {sup 99m}Tc-annexin V (ANX) allows scintigraphic detection of apoptotic cells via specific binding to exposed phosphatidylserine. In myocardial infarction, apoptosis of myocytes is variable and depends especially on the presence or absence of coronary reperfusion. In this study, ANX uptake in non-reperfused experimental myocardial infarcts was compared with uptake of a marker of myocyte necrosis ({sup 111}In-antimyosin antibodies, AM) and an immunohistochemical marker of apoptosis (Apostain). The left anterior coronary artery was ligated in 47 Wistar rats, which were then injected with ANX (n=20), AM (n=21) or both (n=6). Myocardial uptake of ANX and AM was determined at 2 h (n=14), 4 h (n=14) and 24 h (n=19) after coronary ligation (CL), by quantitative autoradiography with (n=23) or without (n=24) gamma imaging. Heart-to-lung ratios (HLRs) and infarct-to-remote myocardium activity ratios (INRs) were calculated on the scintigrams and autoradiograms respectively. Cardiac sections were stained with haematoxylin-eosin and Apostain. The above studies were repeated in 12 normal rats. All rats with CL showed increased ANX and AM uptake in cardiac areas on scintigrams 24 h after CL, with HLRs higher than in controls: 3.1{+-}0.6 versus 1.5{+-}0.3 (p=0.001) for ANX and 1.99{+-}0.44 versus 1.01{+-}0.05 (p<0.0005) for AM. Autoradiography showed intense ANX and AM uptake in infarcts, with comparable topography and INRs at 2 h, 4 h and 24 h after CL (4.6{+-}0.9 versus 5.0{+-}1.8 at 24 h), while Apostain staining was very low (0.06{+-}0.06% of cells). In this model of persistent CL, we observed increased ANX uptake in injured myocardium, comparable in intensity, topography and kinetics to that of AM. There was only minimal Apostain staining in the same areas. (orig.)

  15. Myocardial uptake of {sup 99m}Tc-annexin-V and {sup 111}In-antimyosin-antibodies after ischemia-reperfusion in rats

    Energy Technology Data Exchange (ETDEWEB)

    Sarda-Mantel, Laure [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); AP-HP, Groupe Hospitalier Bichat-Beaujon, Service de Medecine Nucleaire, Paris (France); INSERM, U773, Paris (France); Hopital Bichat, Service de Medecine Nucleaire, Paris (France); Hervatin, Florence [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); CEA, DSV/DRM/SHFJ, Orsay (France); Michel, Jean-Baptiste; Louedec, Liliane [INSERM, U698, Paris (France); Martet, Genevieve [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); INSERM, U773, Paris (France); Rouzet, Francois; Lebtahi, Rachida; Merlet, Pascal; Le Guludec, Dominique [Universite Denis Diderot-Paris 7, UMR S773, Paris (France); AP-HP, Groupe Hospitalier Bichat-Beaujon, Service de Medecine Nucleaire, Paris (France); INSERM, U773, Paris (France); Khaw, Ban-An [Bouve College of Pharmacy and Health Sciences, Center for Drug Targeting and Analysis, Boston, MA (United States)


    Phosphatidylserin exposure on cell surfaces occurs early during apoptosis and is detected in vivo by using {sup 99m}Tc-annexin-V (ANX). Cardiomyocyte membrane disruption is detected in vivo by using {sup 111}In-antimyosin-antibodies (AM). We aimed to determine if ANX and AM allow evaluation of the time-course of these two distinct cell death events after myocardial ischemia-reperfusion. Coronary tying (20 min) followed by reperfusion (IR) was performed in 31 rats. Twelve of the rats were injected with ANX, 11 with AM, and eight with both tracers. Myocardial uptake of tracers was studied 1-2 h, 4 h, or 24 h after IR by scintigraphy (ANX, n = 14) and autoradiography (all cases), and compared to histology and Apostain staining. Scintigraphy was positive in all rats 2 h after IR and in three of five rats at 24 h. On autoradiography, ANX activity was intense in myocardial lesions as early as 1 h post-IR, whereas AM activity was mild at 2 h then increased at 4 h post-IR. ANX and AM uptakes evolved from mid-myocardium to endocardial and epicardial regions from 2 h to 24 h post-IR. Apostain staining was significant in myocardial lesions (p < 10{sup 6} compared to six sham-operated rats). On histology, myocardial lesion was characterized by interstitial oedema, myocytes necrosis, and dramatic thinning at 24 h. These data suggest that ANX and AM allow temporal and regional evaluations of PS exposure and membrane disruption, respectively, during myocytes death after 20-min myocardial ischemia followed by reperfusion. Also, (i) apoptosis starts very early in injured myocardium, (ii) myocyte necrosis occurs later (3-4 h post-reperfusion), and (iii) most dead cells are removed from mid-myocardium between 6 h and 24 h after reperfusion. (orig.)

  16. 2-Deoxyglucose incorporation into rat brain glycogen during measurement of local cerebral glucose utilization by the 2-deoxyglucose method

    Energy Technology Data Exchange (ETDEWEB)

    Nelson, T.; Kaufman, E.E.; Sokoloff, L.


    The incorporation of 14C into glycogen in rat brain has been measured under the same conditions that exist during the measurement of local cerebral glucose utilization by the autoradiographic 2-(14C)deoxyglucose method. The results demonstrate that approximately 2% of the total 14C in brain 45 min after the pulse of 2-(14C)deoxyglucose is contained in the glycogen portion, and, in fact, incorporated into alpha-1-4 and alpha-1-6 deoxyglucosyl linkages. When the brain is removed by dissection, as is routinely done in the course of the procedure of the 2-(14C)deoxyglucose method to preserve the structure of the brain for autoradiography, the portion of total brain 14C contained in glycogen falls to less than 1%, presumably because of postmortem glycogenolysis which restores much of the label to deoxyglucose-phosphates. In any case, the incorporation of the 14C into glycogen is of no consequence to the validity of the autoradiographic deoxyglucose method, not because of its small magnitude, but because 2-(14C)deoxyglucose is incorporated into glycogen via (14C)deoxyglucose-6-phosphate, and the label in glycogen represents, therefore, an additional ''trapped'' product of deoxyglucose phosphorylation by hexokinase. With the autoradiographic 2-(14C)deoxyglucose method, in which only total 14C concentration in the brain tissue is measured by quantitative autoradiography, it is essential that all the labeled products derived directly or indirectly from (14C)deoxyglucose phosphorylation by hexokinase be retained in the tissue; their chemical identity is of no significance.

  17. Distribution of bisphenol A and tetrabromobisphenol A in quail eggs, embryos and laying birds and studies on reproduction variables in adults following in ovo exposure

    Energy Technology Data Exchange (ETDEWEB)

    Halldin, K.; Berg, C.; Brandt, I.; Brunstroem, B. [Department of Environmental Toxicology, Evolutionary Biology Centre, Uppsala Univ. (Sweden); Bergman, Aa. [Department of Environmental Chemistry, Stockholm University, Stockholm (Sweden)


    In this study, uptake and distribution of BPA and tetrabromobisphenol A (TBBPA) in embryos and laying quail were examined as well as variables related to reproduction in adult quail following administration of the compounds into the yolk of embryonated eggs. The uptake of radiolabelled BPA, TBBPA and the reference compound diethylstilboestrol (DES) was studied in the embryos using ss-spectrometry. Autoradiography was employed to examine distribution in egg and embryo after yolk sac injection of BPA or TBBPA and in laying birds, following intravenous and oral administration. Following embryonic exposure to BPA or TBBPA, sexually mature male birds were examined for reproductive behaviour and testis morphology, and females were examined for egg laying and oviduct morphology. Neither BPA (200 {mu}g/g egg) nor TBBPA (15 {mu}g/g egg) caused any significant oestrogen-like effects on the variables studied, although effects on the female oviducts after BPA exposure were indicated. Embryonic exposure to DES is known to cause profound effects on male sexual behaviour and female oviduct morphology at doses 3-5 orders of magnitude lower than the BPA and TBBPA doses used in the present study. The proportions of BPA and TBBPA taken up by the embryos after yolk sac injection were similar to the proportion of DES taken up. Differences in bioavailability, therefore do not account for any major part of the potency differences between DES and the two bisphenol A compounds. The concentration of radioactivity in the embryo, as revealed by autoradiography, was low compared with that in the yolk at all stages studied (days 6, 10 and 15). Pronounced labelling of the bile and the allantoic fluid was observed, however, indicating that both compounds were readily metabolised and excreted. Radiolabelled BPA and TBBPA administered to laying quail were largely excreted via the bile and 9 days after oral dosing, only small amounts of the labelled compound remained within the body. Maternal

  18. 2-(2'-((Dimethylamino)methyl)-4'-(3-[{sup 18}F]fluoropropoxy)-phenylthio)benzenamine for positron emission tomography imaging of serotonin transporters

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Julie L. [Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 (United States); Parhi, Ajit K.; Oya, Shunichi; Lieberman, Brian; Kung, Mei-Ping [Department of Radiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 (United States); Kung, Hank F. [Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 (United States); Department of Radiology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104 (United States)], E-mail:


    Introduction: A new {sup 18}F ligand, 2-(2'-((dimethylamino)methyl)-4'-(3-[{sup 18}F]fluoropropoxy) -phenylthio)benzenamine ([{sup 18}F]1), for positron emission tomography (PET) imaging of serotonin transporters (SERT) was evaluated. Methods: Binding affinity was determined through in vitro binding assays with LLC-PK1 cells overexpressing SERT, NET or DAT (LLC-SERT, LLC-NET and LLC-DAT) and with rat cortical homogenates. Localization and selectivity of [{sup 18}F]1 binding in vivo were evaluated by biodistribution, autoradiography and A-PET imaging studies in rats. Results: This compound displayed excellent binding affinity for SERT in vitro with K{sub i}=0.33 and 0.24 nM in LLC-SERT and rat cortical homogenates, respectively. Biodistribution studies with [{sup 18}F]1 showed good brain uptake (1.61% dose/g at 2 min postinjection), high uptake into the hypothalamus (1.22% dose/g at 30 min) and a high target-to-nontarget (hypothalamus to cerebellum) ratio of 9.66 at 180 min postinjection. Pretreatment with a SERT selective inhibitor considerably inhibited [{sup 18}F]1 binding in biodistribution studies. Ex vivo autoradiography reveals [{sup 18}F]1 localization to brain regions with high SERT density, and this binding was blocked by pretreatment with SERT selective inhibitors. Small animal PET (A-PET) imaging in rats provided clear images of tracer localization in the thalamus, midbrain and striatum. In A-PET chasing experiments, injecting a SERT selective inhibitor 75 min post-tracer injection causes a dramatic reduction in regional radioactivity and the target-to-nontarget ratio. Conclusion: The results of the biological studies and the ease of radiosynthesis with moderately good radiochemical yield (RCY=10-35%) make [{sup 18}F]1 an excellent candidate for SERT PET imaging.

  19. Characterization of a novel acetamidobenzoxazolone-based PET ligand for translocator protein (18 kDa) imaging of neuroinflammation in the brain. (United States)

    Tiwari, Anjani K; Yui, Joji; Fujinaga, Masayuki; Kumata, Katsushi; Shimoda, Yoko; Yamasaki, Tomoteru; Xie, Lin; Hatori, Akiko; Maeda, Jun; Nengaki, Nobuki; Zhang, Ming-Rong


    We developed the novel positron emission tomography (PET) ligand 2-[5-(4-[(11)C]methoxyphenyl)-2-oxo-1,3-benzoxazol-3(2H)-yl]-N-methyl-N-phenylacetamide ([(11)C]MBMP) for translocator protein (18 kDa, TSPO) imaging and evaluated its efficacy in ischemic rat brains. [(11)C]MBMP was synthesized by reacting desmethyl precursor (1) with [(11)C]CH3 I in radiochemical purity of ≥ 98% and specific activity of 85 ± 30 GBq/μmol (n = 18) at the end of synthesis. Biodistribution study on mice showed high accumulation of radioactivity in the TSPO-rich organs, e.g., the lungs, heart, kidneys, and adrenal glands. The metabolite analysis in mice brain homogenate showed 80.1 ± 2.7% intact [(11)C]MBMP at 60 min after injection. To determine the specific binding of [(11)C]MBMP with TSPO in the brain, in vitro autoradiography and PET studies were performed in an ischemic rat model. In vitro autoradiography indicated significantly increased binding on the ipsilateral side compared with that on the contralateral side of ischemic rat brains. This result was supported firmly by the contrast of radioactivity between the ipsilateral and contralateral sides in PET images. Displacement experiments with unlabelled MBMP or PK11195 minimized the difference in uptake between the two sides. In summary, [(11)C]MBMP is a potential PET imaging agent for TSPO and, consequently, for the up-regulation of microglia during neuroinflammation.

  20. Evaluation of CLINDE as potent translocator protein (18 kDa) SPECT radiotracer reflecting the degree of neuroinflammation in a rat model of microglial activation

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    Arlicot, Nicolas; Duval, Stephanie; Guilloteau, Denis; Chalon, Sylvie [Inserm, U930, Tours (France); Universite Francois Rabelais, Tours (France); CHRU de Tours, Tours (France); Katsifis, Andrew; Mattner, Filomena [Australian Nuclear Science and Technology Organisation, Radiopharmaceuticals Research Institute, Sydney (Australia); Garreau, Lucette; Vergote, Jackie; Bodard, Sylvie [Inserm, U930, Tours (France); Universite Francois Rabelais, Tours (France)


    The translocator protein (TSPO; 18 kDa), the new name of the peripheral-type benzodiazepine receptor, is localised in mitochondria of glial cells and expressed in very low concentrations in normal brain. Their expression rises after microglial activation following brain injury. Accordingly, TSPO are potential targets to evaluate neuroinflammatory changes in a variety of CNS disorders. To date, only a few effective tools are available to explore TSPO by SPECT. We characterised here 6-chloro-2-(4'iodophenyl)-3-(N,N-diethyl)-imidazo[1,2-a]pyridine-3-acetamide or CLINDE in a rat model with different stages of excitotoxic lesion. Excitotoxicity was induced in male Wistar rats by unilateral intrastriatal injection of different amounts of quinolinic acid (75, 150 or 300 nmol). Six days later, two groups of rats (n = 5-6/group) were i.v. injected with [{sup 125}I]-CLINDE (0.4 MBq); one group being pre-injected with PK11195 (5 mg/kg). Brains were removed 30 min after tracer injection and the radioactivity of cerebral areas measured. Complementary ex vivo autoradiography, in vitro autoradiography ([{sup 3}H]-PK11195) and immunohistochemical studies (OX-42) were performed on brain sections. In the control group, [{sup 125}I]-CLINDE binding was significantly higher (p < 0.001) in lesioned than that in intact side. This binding disappeared in rats pre-treated with PK11195 (p<0.001), showing specific binding of CLINDE to TSPO. Ex vivo and in vitro autoradiographic studies and immunohistochemistry were consistent with this, revealing a spatial correspondence between radioactivity signal and activated microglia. Regression analysis yielded a positive relation between the ligand binding and the degree of neuroinflammation. These results demonstrate that CLINDE is suitable for TSPO in vivo SPECT imaging to explore their involvement in neurodegenerative disorders associated with microglial activation. (orig.)

  1. The effect of anaesthesia on [{sup 18}F]MK-9470 binding to the type 1 cannabinoid receptor in the rat brain

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    Casteels, Cindy; Van Laere, Koen [KU Leuven and University Hospital Gasthuisberg, Division of Nuclear Medicine, Leuven (Belgium); KU Leuven, MoSAIC, Molecular Small Animal Imaging Center, Leuven (Belgium); Bormans, Guy [KU Leuven, MoSAIC, Molecular Small Animal Imaging Center, Leuven (Belgium); KU Leuven, Laboratory for Radiopharmacy, Leuven (Belgium)


    Small animal PET can be applied to study molecular processes in animal models of a variety of human diseases. In order to keep the animals in a restricted position during imaging, anaesthesia is in many instances inevitable. Using small animal PET and ex vivo autoradiography, we examined the influence of pentobarbital and isoflurane anaesthesia on the rat brain uptake of [{sup 18}F]MK-9470, a radioligand for the type 1 cannabinoid receptor. PET imaging was performed on adult Wistar rats under pentobarbital (n=6) and isoflurane anaesthesia (n=7), and under control conditions (free moving during tracer uptake, n=8). Parametric PET images were generated, anatomically standardized and analysed by voxel-based Statistical Parametric Mapping and a predefined volume of interest approach. Immediately after in vivo PET, brains were processed for ex vivo autoradiography using manually placed regions of interest. An extra group (n=6) was included ex vivo, in which animals were intravenously injected without the use of anaesthetics. Using in vivo and ex vivo molecular imaging techniques, no significant changes in absolute [{sup 18}F]MK-9470 uptake were present in the brain of pentobarbital and isoflurane rats as compared to control conditions. Relative [{sup 18}F]MK-9470 uptake PET values obtained applying global scaling were, however, decreased in the cortex under both anaesthetics (pentobarbital: -13.3{+-}1.4%; isoflurane -8.7 {+-} 3.1%), while an increase was seen in the cerebellum by 13.5 {+-} 4.0% and 13.9 {+-} 4.1% under pentobarbital and isoflurane, respectively. Ex vivo results were in agreement with in vivo findings. These findings suggest a similar, regionally specific interference of pentobarbital and isoflurane anaesthesia with in vivo CB1 receptor imaging using [{sup 18}F]MK-9470. (orig.)

  2. Synthesis and biological evaluation of carbon-11- and fluorine-18-labeled 2-oxoquinoline derivatives for type 2 cannabinoid receptor positron emission tomography imaging

    Energy Technology Data Exchange (ETDEWEB)

    Evens, Nele [Laboratory for Radiopharmacy, K.U. Leuven, 3000 Leuven (Belgium); Muccioli, Giulio G. [Unite de Chimie Pharmaceutique et de Radiopharmacie, U.C. Louvain, 1200 Bruxelles (Belgium); Houbrechts, Nele [Laboratory for Radiopharmacy, K.U. Leuven, 3000 Leuven (Belgium); Lambert, Didier M. [Unite de Chimie Pharmaceutique et de Radiopharmacie, U.C. Louvain, 1200 Bruxelles (Belgium); Verbruggen, Alfons M. [Laboratory for Radiopharmacy, K.U. Leuven, 3000 Leuven (Belgium); Van Laere, Koen [Division of Nuclear Medicine, K.U. Leuven, 3000 Leuven (Belgium); Bormans, Guy M. [Laboratory for Radiopharmacy, K.U. Leuven, 3000 Leuven (Belgium)], E-mail:


    Introduction: The type 2 cannabinoid (CB{sub 2}) receptor is part of the endocannabinoid system and has been suggested as a mediator of several central and peripheral inflammatory processes. Imaging of the CB{sub 2} receptor has been unsuccessful so far. We synthesized and evaluated a carbon-11- and a fluorine-18-labeled 2-oxoquinoline derivative as new PET tracers with high specificity and affinity for the CB{sub 2} receptor. Methods: Two 2-oxoquinoline derivatives were synthesized and radiolabeled with either carbon-11 or fluorine-18. Their affinity and selectivity for the human CB{sub 2} receptor were determined. Biological evaluation was done by biodistribution, radiometabolite and autoradiography studies in mice. Results: In vitro studies showed that both compounds are high affinity CB{sub 2}-specific inverse agonists. Biodistribution study of the tracers in mice showed a high in vivo initial brain uptake and fast brain washout, in accordance with the low CB{sub 2} receptor expression levels in normal brain. A persistently high in vivo binding to the spleen was observed, which was inhibited by pretreatment with two structurally unrelated CB{sub 2} selective inverse agonists. In vitro autoradiography studies with the radioligands confirmed CB{sub 2}-specific binding to the mouse spleen. Conclusion: We synthesized two novel CB{sub 2} receptor PET tracers that show high affinity/selectivity for CB{sub 2} receptors. Both tracers show favourable characteristics as radioligands for central and peripheral in vivo visualization of the CB{sub 2} receptor and are promising candidates for primate and human CB{sub 2} PET imaging.

  3. In Vitro and In Vivo Characterization of Selected Fluorine-18 Labeled Radioligands for PET Imaging of the Dopamine D3 Receptor

    Directory of Open Access Journals (Sweden)

    Natascha Nebel


    Full Text Available Cerebral dopamine D3 receptors seem to play a key role in the control of drug-seeking behavior. The imaging of their regional density with positron emission tomography (PET could thus help in the exploration of the molecular basis of drug addiction. A fluorine-18 labeled D3 subtype selective radioligand would be beneficial for this purpose; however, as yet, there is no such tracer available. The three candidates [18F]1, [18F]2a and [18F]2b were chosen for in vitro and in vivo characterization as radioligands suitable for selective PET imaging of the D3 receptor. Their evaluation included the analysis of radiometabolites and the assessment of non-specific binding by in vitro rat brain autoradiography. While [18F]1 and [18F]2a revealed high non-specific uptake in in vitro rat brain autoradiography, the D3 receptor density was successfully determined on rat brain sections (n = 4 with the candidate [18F]2b offering a Bmax of 20.38 ± 2.67 pmol/g for the islands of Calleja, 19.54 ± 1.85 pmol/g for the nucleus accumbens and 16.58 ± 1.63 pmol/g for the caudate putamen. In PET imaging studies, the carboxamide 1 revealed low signal/background ratios in the rat brain and relatively low uptake in the pituitary gland, while the azocarboxamides [18F]2a and [18F]2b showed binding that was blockable by the D3 receptor ligand BP897 in the ventricular system and the pituitary gland in PET imaging studies in living rats.

  4. Electrophysiological and autoradiographical evidence of V1 vasopressin receptors in the lateral septum of the rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Raggenbass, M.; Tribollet, E.; Dreifuss, J.J.


    Extracellular recordings were obtained from single neurons located in the lateral septum, an area known to receive a vasopressinergic innervation in the rat brain. Approximately half of the neurons tested responded to 8-L-arginine vasopressin (AVP) by a marked increase in firing rate at concentrations greater than 1 nM. The effect of vasopressin was blocked by synthetic structural analogues possessing antagonistic properties on peripheral vasopressin and oxytocin receptors. Oxytocin was much less potent than vasopressin in firing septal neurons, and a selective oxytocic agonist was totally ineffective. The action of vasopressin on neuronal firing was mimicked by the vasopressor agonist (2-phenylalanine,8-ornithine)vasotocin but not by the selective antidiuretic agonist 1-deamino(8-D-arginine)vasopressin. In a parallel study, sites that bind (/sup 3/H)AVP at low concentration (1.5 nM) were found by in vitro autoradiography in the lateral septum. Adjacent sections were also incubated with 1.5 mM (/sup 3/H)AVP and, in addition, with 100 nM (2-phenylalanine,8-ornithine)vasotocin or 1-deamino(8-D-arginine)vasopressin--i.e., the same compounds as those used for the electrophysiological study. Results showed that the vasopressor agonist, but not the antidiuretic agonist, displaced (/sup 3/H)AVP, thus indicating that the vasopressin binding sites detected by autoradiography in the septum were V1 (vasopressor type) rather than V2 (antidiuretic type) receptors. Based on the electrophysiological evidence, we conclude that these receptors, when occupied, lead to increased firing of lateral septal neurons.

  5. Imaging brain inflammation with [(11)C]PK11195 by PET and induction of the peripheral-type benzodiazepine receptor after transient focal ischemia in rats. (United States)

    Rojas, Santiago; Martín, Abraham; Arranz, Maria J; Pareto, Deborah; Purroy, Jesús; Verdaguer, Esther; Llop, Jordi; Gómez, Vanessa; Gispert, Joan D; Millán, Olga; Chamorro, Angel; Planas, Anna M


    [(11)C]PK11195 is used in positron emission tomography (PET) studies for imaging brain inflammation in vivo as it binds to the peripheral-type benzodiazepine receptor (PBR) expressed by reactive glia and macrophages. However, features of the cellular reaction required to induce a positive [(11)C]PK11195 signal are not well characterized. We performed [(11)C]PK11195 PET and autoradiography in rats after transient focal cerebral ischemia. We determined [(3)H]PK11195 binding and PBR expression in brain tissue and examined the lesion with several markers. [(11)C]PK11195 standard uptake value increased at day 4 and grew further at day 7 within the ischemic core. Accordingly, ex vivo [(3)H]PK11195 binding increased at day 4, and increases further at day 7. The PET signal also augmented in peripheral regions, but to a lesser extent than in the core. Binding in the region surrounding infarction was supported by [(11)C]PK11195 autoradiography at day 7 showing that the radioactive signal extended beyond the infarcted core. Enhanced binding was preceded by increases in PBR mRNA expression in the ipsilateral hemisphere, and a 18-kDa band corresponding to PBR protein was detected. Peripheral-type benzodiazepine receptor immunohistochemistry showed subsets of ameboid microglia/macrophages within the infarcted core showing a distinctive strong PBR expression from day 4. These cells were often located surrounding microhemorrhages. Reactive astrocytes forming a rim surrounding infarction at day 7 also showed some PBR immunostaining. These results show cellular heterogeneity in the level of PBR expression, supporting that PBR is not a simple marker of inflammation, and that the extent of [(11)C]PK11195 binding depends on intrinsic features of the inflammatory cells.

  6. {sup 18}F-FDG and {sup 18}F-FET uptake in experimental soft tissue infection

    Energy Technology Data Exchange (ETDEWEB)

    Kaim, Achim H.; Weber, Bruno; Schulthess, Gustav K. von; Buck, Alfred [Nuclear Medicine, University Hospital Zurich (Switzerland); Kurrer, Michael O. [Department of Pathology, University Hospital Zurich (Switzerland); Westera, Gerrit [Center for Radiopharmaceutical Sciences, University Hospital Zuerich (Switzerland); Schweitzer, Alain [Novartis Pharma Inc., Basel (Switzerland); Gottschalk, Jochen [Institute of Medical Microbiology, University Hospital Zurich (Switzerland)


    The aim of this study was to compare the uptake of {sup 18}F-fluoroethyl-L-tyrosine ({sup 18}F-FET) with that of {sup 18}F-fluorodeoxyglucose ({sup 18}F-FDG) in activated inflammatory white blood cells. Unilateral thigh muscle abscesses were induced in 11 rats by intramuscular inoculation of 0.1 ml of a bacterial suspension (S. aureus, 1.2 x 10{sup 9} CFU/ml). Four animals were intraperitoneally injected with 130-180 MBq {sup 18}F-FDG, four with 140-170 MBq {sup 18}F-FET and three with a mixture of 140-170 MBq {sup 18}F-FET and 1.8 MBq {sup 14}C-deoxyglucose. Autoradiography (10 {mu}m slice thickness) of the abscess and the contralateral muscle was performed and detailed spatial correlation of autoradiography and histopathology (haematoxylin-eosin staining) was obtained. Regions of interest were placed on the abscess wall and the grey values (digitised image intensities) measured were converted to kBq/cc per kBq injected activity per gram (SUV). Areas with increased {sup 18}F-FDG uptake corresponded to cellular inflammatory infiltrates mainly consisting of granulocytes. The SUV was calculated to be 4.08{+-}0.65 (mean{+-}SD). The uptake of {sup 18}F-FET in activated white blood cells was not increased: the SUV of the abscess wall, at 0.74{+-}0.14, was even below that of contralateral muscle. The low uptake of {sup 18}F-FET in non-neoplastic inflammatory cells promises a higher specificity for the detection of tumour cells than is achieved with {sup 18}F-FDG, since the immunological host response will not be labelled and inflammation can be excluded. (orig.)

  7. Micro–Positron Emission Tomography/Contrast-Enhanced Computed Tomography Imaging of Orthotopic Pancreatic Tumor–Bearing Mice Using the αvβ3 Integrin Tracer 64Cu-Labeled Cyclam-RAFT-c(-RGDfK-4

    Directory of Open Access Journals (Sweden)

    Winn Aung


    Full Text Available The purpose of this study was to develop a clinically relevant orthotopic xenotransplantation model of pancreatic cancer and to perform a preclinical evaluation of a new positron emission tomography (PET imaging probe, 64Cu-labeled cyclam-RAFT-c(-RGDfK-4 peptide (64Cu-RAFT-RGD, using this model. Varying degrees of αvβ3 integrin expression in several human pancreatic cancer cell lines were examined by flow cytometry and Western blotting. The cell line BxPC-3, which is stably transfected with a red fluorescence protein (RFP, was used for surgical orthotopic implantation. Orthotopic xenograft was established in the pancreas of recipient nude mice. An in vivo probe biodistribution and receptor blocking study, preclinical PET imaging coregistered with contrast-enhanced computed tomography (CECT comparing 64Cu-RAFT-RGD and 18F-fluoro-2-deoxy-D-glucose (18F-FDG accumulation in tumor, postimaging autoradiography, and histologic and immunohistochemical examinations were done. Biodistribution evaluation with a blocking study confirmed that efficient binding of probe to tumor is highly αvβ3 integrin specific. 64Cu-RAFT-RGD PET combined with CECT provided for precise and easy detection of cancer lesions. Autoradiography, histologic, and immunohistochemical examinations confirmed the accumulation of 64Cu-RAFT-RGD in tumor versus nontumor tissues. In comparative PET studies, 64Cu-RAFT-RGD accumulation provided better tumor contrast to background than 18F-FDG. Our results suggest that 64Cu-RAFT-RGD PET imaging is potentially applicable for the diagnosis of αvβ3 integrin–expressing pancreatic tumors.

  8. Evaluation of [{sup 11}C]rofecoxib as PET tracer for cyclooxygenase 2 overexpression in rat models of inflammation

    Energy Technology Data Exchange (ETDEWEB)

    Vries, Erik F.J. de [Department of Nuclear Medicine and Molecular Imaging, University Medical Center Groningen, University of Groningen, P.O. Box 30.001, 9700 RB Groningen (Netherlands)], E-mail:; Doorduin, Janine; Dierckx, Rudi A.; Waarde, Aren van [Department of Nuclear Medicine and Molecular Imaging, University Medical Center Groningen, University of Groningen, P.O. Box 30.001, 9700 RB Groningen (Netherlands)


    Background: Overexpression of cyclooxygenase type 2 (COX-2) is triggered by inflammatory stimuli, but it also plays a prominent role in the initiation and progression of various diseases. This study aims to investigate [{sup 11}C]rofecoxib as a positron emission tomography (PET) tracer for COX-2 expression. Methods: [{sup 11}C]Rofecoxib was prepared by methylation of its sulphinate precursor. Regional brain distribution and specific binding of [{sup 11}C]rofecoxib in healthy rats was studied by ex vivo biodistribution and autoradiography. Regional brain distribution and PET imaging studies were also performed on rats with severe encephalitis, caused by nasal infection with herpes simplex virus (HSV). Finally, ex vivo biodistribution and blocking studies were carried in rats with a sterile inflammation, induced by intramuscular turpentine injection. Results: [{sup 11}C]rofecoxib brain uptake in control animals corresponded with the known distribution of COX-2. Pretreatment with NS398 significantly reduced tracer uptake in the cingulate/frontopolar cortex, whereas the reduction in hippocampus approached significance. Ex vivo autoradiography also revealed preferential tracer uptake in hippocampus and cortical areas that could be blocked by NS398. In HSV-infected animals, [{sup 11}C]rofecoxib uptake was moderately increased in all brain regions, but it could not be blocked with indomethacin. Yet, some PET images revealed increased tracer uptake in brain areas with microglia activation. In turpentine-injected animals, [{sup 11}C]rofecoxib uptake in inflamed muscle was not higher than in control muscle and could not be blocked with NS398. Indomethacin caused a slight reduction in muscle uptake. Conclusions: Despite the apparent correlation between [{sup 11}C]rofecoxib uptake and COX-2 distribution in healthy rats, [{sup 11}C]rofecoxib could not unambiguously detect COX-2 overexpression in two rat models of inflammation.

  9. Region specific up-regulation of oxytocin receptors in the opioid Oprm1-/- mouse model of autism

    Directory of Open Access Journals (Sweden)

    Valentina eGigliucci


    Full Text Available Autism spectrum disorders (ASDs are characterized by impaired communication, social impairments and restricted and repetitive behaviors and interests. Recently altered motivation and reward processes have been suggested to participate in the physiopathology of ASDs, and μ-opioid receptors (MORs have been investigated in relation to social reward due to their involvement in the neural circuitry of reward. Mice lacking a functional MOR gene (Oprm1-/- mice display abnormal social behavior and major autistic-like core symptoms, making them an animal model of autism. The oxytocin (OXT system is a key regulator of social behavior and co-operates with the opioidergic system in the modulation of social behavior. To better understand the opioid-OXT interplay in the central nervous system, we first determined the expression of the oxytocin receptor (OXTR in the brain of WT C57BL6/J mice by quantitative autoradiography; we then evaluated OXTR regional alterations in Oprm1-/- mice. Moreover, we tested these mice in a paradigm of social behavior, the male-female social interaction test, and analyzed the effects of acute intranasal OXT treatment on their performance. In autoradiography, Oprm1-/- mice selectively displayed increased OXTR expression in the Medial Anterior Olfactory Nucleus, the Central and Medial Amygdaloid nuclei and the Nucleus Accumbens. Our behavioral results confirmed that Oprm1-/- male mice displayed social impairments, as indicated by reduced ultrasonic calls, and that these were rescued by a single intranasal administration of OXT. Taken together, our results provide evidence of an interaction between OXT and opioids in socially relevant brain areas and in the modulation of social behavior. Moreover, they suggest that the oxytocinergic system may act as a compensative mechanism to bypass and/or restore alterations in circuits linked to impaired social behavior.

  10. The effects of ionizing radiation in the rat's mandibular bone freeding the hypernomic calcium-deficient diet

    Energy Technology Data Exchange (ETDEWEB)

    Hasegawa, Gen; Kurita, Akihiko; Nasu, Masanori; Furumoto, Keiichi (Nippon Dental Univ., Tokyo (Japan))


    The mandibles of rats in a group maintained on the Ca-deficient diet for a long period were irradiated with 30 Gy. To study the effects of radiation, serum Ca and inorganic phosphorus levels were determined for 3 weeks, and the data were compared with findings obtained from rats maintained on a standard diet by autoradiography using [sup 45]Ca and microradiography. The serum Ca level tended to decrease with time after irradiation in the irradiated group maintained on the Ca-deficient diet, but there was no significant difference between the group maintained on the Ca-deficient diet and the group maintained on the standard diet. The serum inorganic phosphorus levels were almost constant throughout the observation period in both the non-irradiated and radiated groups regardless of diet. Uptake of [sup 45]Ca was examined by autoradiography. Both the non-irradiated and irradiated groups maintained on the Ca-deficient diet showed intense [sup 45]Ca uptake, there was almost no difference between these groups in photographic density or in weekly changes after irradiation. The microradiographic study of bone trabeculae revealed only slight changes in the bone cortex after irradiation in the group maintained on the standard diet. On day 3 after irradiation both thinning and roughness of the trabeculae were observed in the interradicular septa and incisal inferior margin and on day 7 in cancellous bone. In the groups maintained on the Ca-deficient diet, marked thinning and roughness of the trabeculae were observed mainly in the cancellous bone. (author).

  11. Imaging brain neuronal activity using functionalized magnetonanoparticles and MRI. (United States)

    Akhtari, Massoud; Bragin, Anatol; Moats, Rex; Frew, Andrew; Mandelkern, Mark


    This study explored the use of non-radioactive 2-deoxy glucose (2DG)-labeled magnetonanoparticles (MNP) and magnetic resonance imaging (MRI) to detect functional activity during rest, peripheral stimulation, and epileptic seizures, in animal models. Non-radioactive 2DG was covalently attached to magnetonanoparticles composed of iron oxide and dextran and intravenous (tail) injections were performed. 2DG-MNP was injected in resting and stimulated naïve rodents and the subsequent MRI was compared to published (14)C-2DG autoradiography data. Reproducibility and statistical significance was established in one studied model. Negative contrast enhancement (NCE) in acute seizures and chronic models of epilepsy were investigated. MRI NCE due to 2DG-MNP particles was compared to that of plain (unconjugated) MNP in one animal. NCE due to 2DG-MNP particles at 3 T, which is approved for human use, was also investigated. Histology showed presence of MNP (following intravenous injection) in the brain tissues of resting naïve animal. 2DG-MNP intraparenchymal uptake was visible on MRI and histology. The locations of NCE agreed with published results of 2DG autoradiography in resting and stimulated animals and epileptic rats. Localization of epileptogenicity was confirmed by subsequent depth-electrode EEG (iEEG). Non-radioactive 2DG-MNP can cross the blood-brain barrier (BBB) and may accurately localize areas of increased activity. Although, this proof-of-principle study involves only a limited number of animals, and much more research and quantification are necessary to demonstrate that 2DG-MNP, or MNPs conjugated with other ligands, could eventually be used to image localized cerebral function with MRI in humans, this MNP-MRI approach is potentially applicable to the use of many bioactive molecules as ligands for imaging normal and abnormal localized cerebral functions.

  12. Repeated dexamphetamine treatment alters the dopaminergic system and increases the phMRI response to methylphenidate (United States)

    Schrantee, Anouk; Tremoleda, Jordi L.; Wylezinska-Arridge, Marzena; Bouet, Valentine; Hesseling, Peter; Meerhoff, Gideon F.; de Bruin, Kora M.; Koeleman, Jan; Freret, Thomas; Boulouard, Michel; Desfosses, Emilie; Galineau, Laurent; Gozzi, Alessandro; Dauphin, François; Gsell, Willy; Booij, Jan; Lucassen, Paul J.; Reneman, Liesbeth


    Dexamphetamine (AMPH) is a psychostimulant drug that is used both recreationally and as medication for attention deficit hyperactivity disorder. Preclinical studies have demonstrated that repeated exposure to AMPH can induce damage to nerve terminals of dopamine (DA) neurons. We here assessed the underlying neurobiological changes in the DA system following repeated AMPH exposure and pre-treated rats with AMPH or saline (4 times 5 mg/kg s.c., 2 hours apart), followed by a 1-week washout period. We then used pharmacological MRI (phMRI) with a methylphenidate (MPH) challenge, as a sensitive and non-invasive in-vivo measure of DAergic function. We subsequently validated the DA-ergic changes post-mortem, using a.o. high-performance liquid chromatography (HPLC) and autoradiography. In the AMPH pre-treated group, we observed a significantly larger BOLD response to the MPH challenge, particularly in DA-ergic brain areas and their downstream projections. Subsequent autoradiography studies showed that AMPH pre-treatment significantly reduced DA transporter (DAT) density in the caudate-putamen (CPu) and nucleus accumbens, whereas HPLC analysis revealed increases in the DA metabolite homovanillic acid in the CPu. Our results suggest that AMPH pre-treatment alters DAergic responsivity, a change that can be detected with phMRI in rats. These phMRI changes likely reflect increased DA release together with reduced DAT binding. The ability to assess subtle synaptic changes using phMRI is promising for both preclinical studies of drug discovery, and for clinical studies where phMRI can be a useful tool to non-invasively investigate DA abnormalities, e.g. in neuropsychiatric disorders. PMID:28241065

  13. Survey of the chemical diffusion at infinite dilution in the nickel-plutonium and aluminium-uranium systems; Contribution a l'etude de l'heterodiffusion a dilution infinie systemes nickel-plutonium et aluminium-uranium

    Energy Technology Data Exchange (ETDEWEB)

    Blechet, J.J. [Commissariat a l' Energie Atomique, Fontenay-aux-Roses (France). Centre d' Etudes Nucleaires


    Solubility S{sub 0} and chemical diffusion coefficients D{sub PuNi} at infinite dilution of plutonium in nickel have been determined by autoradiography {alpha} in poly-phased system by the welded couples method: S{sub 0} varies from 40 to 80.10{sup -6} (atomic concentration) and D{sub PuNi} follows an Arrhenius law D = D{sub 0} exp (-Q/RT) with 0.03 cm{sup 2}/s < D{sub 0} < 1.6 cm{sup 2}/s and 46000 cal/mole < Q < 56000 cal/mole. Diffusion of uranium in aluminium have been carried out by fissiography using the thin layer method. Frequency factor lies between 0.01 and 3.1 cm{sup 2}/s and the activation energy lies between 24000 and 34000 cal/mole. (author) [French] La solubilite S{sub 0} et les coefficients de diffusion chimique D{sub PuNi}, a dilution infinie, du plutonium dans le nickel ont ete determines par autoradiographie {alpha} sur des couples soudes en systeme polyphase. Entre 1000 et 1125 deg. C. S{sub 0} varie de 40 a 80.10{sup -6} et D obeit a une loi d'ARRHENIUS (concentration atomique) D = D{sub 0} exp (-Q/RT) avec 0.03 cm{sup 2}s{sup -1} < D{sub 0} < 1.60 cm{sup 2}s{sup -1} 46000 calories par mole < Q < 56000 calories par mole. La diffusion de l'uranium dans l'aluminium a ete etudiee par fissiographie en utilisant la technique du depot mince. Le facteur de frequence est situe entre 0.01 et 3.1 cm{sup 2}s{sup -1} et l'energie d'activation entre 24000 et 34000 calories par mole. (auteur)

  14. Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA. (United States)

    Klein, A B; Bay, T; Villumsen, I S; Falk-Petersen, C B; Marek, A; Frølund, B; Clausen, R P; Hansen, H D; Knudsen, G M; Wellendorph, P


    GHB (γ-hydroxybutyric acid) is a compound endogenous to mammalian brain with high structural resemblance to GABA. GHB possesses nanomolar-micromolar affinity for a unique population of binding sites, but the exact nature of these remains elusive. In this study we utilized the highly selective GHB analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations, (3)H-HOCPCA displays excellent signal-to-noise ratios using rodent brain autoradiography, which makes it a valuable ligand for anatomical quantification of native GHB binding site levels. Our data confirmed that (3)H-HOCPCA labels only the high-affinity specific GHB binding site, found in high density in cortical and hippocampal regions. The experiments revealed markedly stronger binding at pH 6.0 (Kd 73.8 nM) compared to pH 7.4 (Kd 2312 nM), as previously reported for other GHB radioligands but similar Bmax values. Using (3)H-HOCPCA we analyzed the GHB binding protein profile during mouse brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over the commonly used radioligand (3)H-NCS-382, and thus a very suitable in vitro tool for qualitative and quantitative autoradiography of the GHB high-affinity site.

  15. Increased brain dopamine D4-like binding after chronic ethanol is not associated with behavioral sensitization in mice. (United States)

    Quadros, Isabel Marian Hartmann; Nobrega, Jose Nascimento; Hipolide, Debora Cristina; Souza-Formigoni, Maria Lucia Oliveira


    Dopaminergic D4 receptors have been hypothesized to be involved in neuropsychiatric disorders and substance abuse. In mice, repeated ethanol administration may induce behavioral sensitization, a phenomenon of increased sensitivity to the drug's stimulant properties. This study aimed to analyze brain D4 receptors binding in mice with different levels of behavioral sensitization to ethanol. Male Swiss mice received 2.2 g/kg ethanol (n = 64) or saline (n = 16) intraperitoneally daily for 21 days and were weekly tested for locomotor activity and for blood ethanol levels. According to the locomotor scores presented across test days, ethanol-treated mice were classified as "sensitized" or "nonsensitized." Twenty-four hours after the last administration, mice were sacrificed and brains were processed for autoradiography. Brain D4 binding was assessed by quantitative autoradiography using [3H]nemonapride + raclopride in three groups: saline-treated controls (n = 10), ethanol-sensitized (n = 11), and ethanol-nonsensitized (n = 9) mice. Both sensitized and nonsensitized mice showed higher D4 binding densities than saline-treated controls in the posterior caudate-putamen and the olfactory tubercle (p < .02), but only sensitized mice presented higher D4 binding than controls at the lateral septal nucleus (p < .02). However, there were no differences between sensitized and nonsensitized mice in any of the brain regions analyzed. Furthermore, sensitized and nonsensitized mice presented similar blood ethanol levels during the treatment. The higher D4 binding levels observed in both ethanol-treated subgroups (sensitized and nonsensitized) suggest that chronic ethanol treatment may induce upregulation of D4 receptors in specific brain regions. However, this mechanism does not seem to be associated with the differential ability to develop behavioral sensitization to ethanol in mice.

  16. Autoradiographic visualization of angiotensin-converting enzyme in rat brain with (/sup 3/H)captopril: localization to a striatonigral pathway

    Energy Technology Data Exchange (ETDEWEB)

    Strittmatter, S.M.; Lo, M.M.S.; Javitch, J.A.; Snyder, S.H.


    The authors have visualized angiotensin-converting enzyme (ACE; dipeptidyl carboxypeptidase, peptidylpeptide hydrolase, EC in rat brain by in vitro (/sup 3/H)captopril autoradiography. (/sup 3/H)Captopril binding to brain slices displays a high affinity (K/sub d/ = 1.8 x 10/sup -9/ M) and a pharmacological profile similar to that of ACE activity. Very high densities of (/sup 3/H)captopril binding were found in the choroid plexus and the subfornical organ. High densities were present in the caudate putamen and substantia nigra, zona reticulata. Moderate levels were found in the entopeduncular nucleus, globus pallidus, and median eminence of the hypothalamus. Lower levels were detectable in the supraoptic and paraventricular nuclei of the hypothalamus, the media habenula, the median preoptic area, and the locus coeruleus. Injection of ibotenic acid or colchicine into the caudate putamen decreased (/sup 3/H)captopril-associated autoradiographic grains by 85% in the ipsilateral caudate putamen and by > 50% in the ipsilateral substantia nigra. Thus, ACE in the substantia nigra is located on presynaptic terminals of axons originating from the caudate putamen, and ACE in the caudate putamen is situated in neuronal perikarya or at the terminals of striatal interneurons. The lack of effect of similar injections into the substantia nigra confirmed that the caudate putamen injections did not cause trans-synaptic changes. The presence of (/sup 3/H)captopril binding is consistent with an ACE-mediated production of angiotensin II in some brain regions. Although (/sup 3/H)captopril autoradiography reveals ACE in a striatonigral pathway, there is no evidence for angiotensin II involvement in such a neuronal pathway. 26 references, 4 figures, 2 tables.

  17. Metabolic activation of phenacetin in rat nasal mucosa: dose-dependent binding to the glands of Bowman

    Energy Technology Data Exchange (ETDEWEB)

    Brittebo, E.B.


    The metabolism and binding of the analgetic drug (ring-/sup 3/H)phenacetin in the nasal mucosa were studied in vitro and in vivo in male Sprague-Dawley rats. As shown by whole-body and light microscopic autoradiography there was an irreversible binding of metabolites to the glands of Bowman in the olfactory mucosa after high but not after low doses of (/sup 3/H)phenacetin. In the other tissues, the distribution of radioactivity was not changed when the dose was increased. Autoradiography of (/sup 3/H)-acetaminophen showed no preferential uptake of radioactivity in the olfactory mucosa. At incubation of nasal septa with (/sup 3/H)phenacetin in vitro, a binding of metabolites to the glands of Bowman was observed indicating that the metabolism occurred in situ. In rats, glutathione (GSH) depleted by pretreatment with phorone, there was a binding to the glands of Bowman in the olfactory mucosa also after a trace dose of (/sup 3/H)phenacetin. Addition of GSH decreased the irreversible binding of (/sup 3/H)phenacetin metabolites that occurred in 9000 X g nasal mucosa supernatants incubated with (/sup 3/H)phenacetin. There was a moderate decrease in the level of nonprotein sulfhydryl groups, mainly GSH, in the olfactory mucosa after administration of 100-300 mg/kg phenacetin. Collectively, these data suggest that phenacetin is metabolized and subsequent to GSH depletion, bound preferentially in the glands of Bowman. The data also suggest that in situ metabolic activation and binding of phenacetin in the rat nasal mucosa at high doses may play a role in the pathogenesis of the nasal tumors induced by high doses of phenacetin in the rat.

  18. Effect of Iboga alkaloids on µ-opioid receptor-coupled G protein activation.

    Directory of Open Access Journals (Sweden)

    Tamara Antonio

    Full Text Available OBJECTIVE: The iboga alkaloids are a class of small molecules defined structurally on the basis of a common ibogamine skeleton, some of which modify opioid withdrawal and drug self-administration in humans and preclinical models. These compounds may represent an innovative approach to neurobiological investigation and development of addiction pharmacotherapy. In particular, the use of the prototypic iboga alkaloid ibogaine for opioid detoxification in humans raises the question of whether its effect is mediated by an opioid agonist action, or if it represents alternative and possibly novel mechanism of action. The aim of this study was to independently replicate and extend evidence regarding the activation of μ-opioid receptor (MOR-related G proteins by iboga alkaloids. METHODS: Ibogaine, its major metabolite noribogaine, and 18-methoxycoronaridine (18-MC, a synthetic congener, were evaluated by agonist-stimulated guanosine-5´-O-(γ-thio-triphosphate ([(35S]GTPγS binding in cells overexpressing the recombinant MOR, in rat thalamic membranes, and autoradiography in rat brain slices. RESULTS AND SIGNIFICANCE: In rat thalamic membranes ibogaine, noribogaine and 18-MC were MOR antagonists with functional Ke values ranging from 3 uM (ibogaine to 13 uM (noribogaine and 18MC. Noribogaine and 18-MC did not stimulate [(35S]GTPγS binding in Chinese hamster ovary cells expressing human or rat MORs, and had only limited partial agonist effects in human embryonic kidney cells expressing mouse MORs. Ibogaine did not did not stimulate [(35S]GTPγS binding in any MOR expressing cells. Noribogaine did not stimulate [(35S]GTPγS binding in brain slices using autoradiography. An MOR agonist action does not appear to account for the effect of these iboga alkaloids on opioid withdrawal. Taken together with existing evidence that their mechanism of action also differs from that of other non-opioids with clinical effects on opioid tolerance and withdrawal, these

  19. Evaluation of a radiolabelled peripheral benzodiazepine receptor ligand in the central nervous system inflammation of experimental autoimmune encephalomyelitis: a possible probe for imaging multiple sclerosis

    Energy Technology Data Exchange (ETDEWEB)

    Mattner, F.; Katsifis, A.; Ballantyne, P. [ANSTO, Radiopharmaceuticals Division, Lucas Heights (Australia); Staykova, M.; Willenborg, D.O. [Australian National University Medical School, The Canberra Hospital, Neurosciences Research Unit, Woden, Canberra (Australia)


    Peripheral benzodiazepine receptors (PBRs) are upregulated on macrophages and activated microglia, and radioligands for the PBRs can be used to detect in vivo neuroinflammatory changes in a variety of neurological insults, including multiple sclerosis. Substituted 2-phenyl imidazopyridine-3-acetamides with high affinity and selectivity for PBRs have been prepared that are suitable for radiolabelling with a number of positron emission tomography and single-photon emission computed tomography (SPECT) isotopes. In this investigation, the newly developed high-affinity PBR ligand 6-chloro-2-(4'-iodophenyl)-3-(N,N-diethyl)imidazo[1,2-a]pyridine-3-acetamide, or CLINDE, was radiolabelled with{sup 123}I and its biodistribution in the central nervous system (CNS) of rats with experimental autoimmune encephalomyelitis (EAE) evaluated. EAE was induced in male Lewis rats by injection of an emulsion of myelin basic protein and incomplete Freund's adjuvant containing Mycobacterium butyricum. Biodistribution studies with{sup 123}I-CLINDE were undertaken on EAE rats exhibiting different clinical disease severity and compared with results in controls. Disease severity was confirmed by histopathology in the spinal cord of rats. The relationship between inflammatory lesions and PBR ligand binding was investigated using ex vivo autoradiography and immunohistochemistry on rats with various clinical scores. {sup 123}I-CLINDE uptake was enhanced in the CNS of all rats exhibiting EAE when compared to controls. Binding reflected the ascending nature of EAE inflammation, with lumbar/sacral cord > thoracic cord > cervical cord > medulla. The amount of ligand binding also reflected the clinical severity of disease. Ex vivo autoradiography and immunohistochemistry revealed a good spatial correspondence between radioligand signal and foci of inflammation and in particular ED-1{sup +} cells representing macrophages and microglia. These results demonstrate the ability of {sup 123}I

  20. Evaluation of C.L.I.N.D.E. as potent peripheral-type benzodiazepine receptor tracer in a rat model of micro-glial activation

    Energy Technology Data Exchange (ETDEWEB)

    Arlicot, N.; Guilloteau, D.; Chalon, S. [Institut National de la Sante et de la Recherche Medicale (INSERM), U619, 37 - Tours (France); Universite Francois Rabelais de Tours, 37 (France); Katsifis, A.; Mattner, F. [ANSTO, Sydney (Australia)


    The peripheral-type benzodiazepine receptors (P.B.R.) are localized in mitochondria of glial cells and are very low expressed in normal brain. Their expression rises after micro-glial activation consecutive to brain injury. Accordingly, P.B.R. are potential targets to evaluate neuro inflammatory changes in a variety of C.N.S. disorders. To date no effective tool is available to explore P.B.R. by SPECT. We characterized here 6-chloro-2-(4 iodophenyl)-3-(N,N-diethyl)-imidazo[1,2-a]pyridine- 3-acetamide, C.L.I.N.D.E., in a rat model of excitotoxic lesion. Excitotoxicity was induced in male Wistar rats by unilateral intra striatal injection of different amounts of quinolinic acid (Q.A.: 75, 150 or 300 nmol). One week later, 2 groups of rats (n = 5-6/group) were i.v. injected with [{sup 125}I]-C.L.I.N.D.E. (0.4 MBq), one group being pre-injected with P.K.11195 (5 mg/kg). Brains were removed 30 min after tracer injection and the radioactivity of cerebral areas measured. Complementary ex vivo autoradiography and immunohistochemical studies using O.X.-42 were performed on brain sections In the control group, [{sup 125}I]-C.L.I.N.D.E. binding was significantly higher ( p < 0.001) in lesioned than that in intact side (striatum: 0.552 {+-} 0.109 vs. 0.123 {+-} 0.012% I.D./g tissue; cortex: 0.385 {+-} 0.126 vs. 0.131 {+-} 0.007% with 300 nmol Q.A.). This binding disappeared in rats pretreated with P.K.11195 ( p < 0.001), showing specific binding of C.L.I.N.D.E. to P.B.R.. Ex vivo autoradiography and immunohistochemistry were consistent with this, revealing a spatial correspondence between radioactivity signal and activated micro-glia. Regression analysis yielded a significant correlation ( p < 0.001) between the ligand binding and the dose of Q.A.. These results demonstrate that C.L.I.N.D.E. is suitable for P.B.R. in vivo SPECT imaging to explore their involvement in neuro degenerative disorders associated with micro-glial activation. (authors)

  1. Unscheduled DNA synthesis in rat pleural mesothelial cells treated with mineral fibres. (United States)

    Renier, A; Lévy, F; Pillière, F; Jaurand, M C


    Unscheduled DNA synthesis (UDS) was studied in confluent rat pleural mesothelial cells (RPMCs) arrested in G0/G1 with hydroxyurea (HU) and treated with various fibre types, i.e., chrysotile, crocidolite or attapulgite. In addition, the effects of UV light and of benzo[a]pyrene were determined as references. Using autoradiography after [3H]thymidine incorporation ([3H]dThd), RPMCs treated with 4 micrograms/cm2 of chrysotile fibres exhibited a low but significant enhancement of net grains compared to untreated cells. Treatment with higher doses of chrysotile was not possible because of the impairment of microscopic observation due to the presence of the fibres. Using liquid scintillation counting, RPMCs treated with chrysotile or crocidolite showed a significant dose-dependent increase in [3H]dThd incorporation compared to untreated cells. In contrast, attapulgite did not enhance [3H]dThd incorporation compared to untreated cells. Treatment of RPMCs with 1, 2 or 4 micrograms/ml of benzo[a]pyrene resulted in a significant increase in [3H]dThd incorporation. In order to discount a possible role of S cells in the augmentation of [3H]dThd incorporation, despite the presence of 5 mM HU, S cells were counted by autoradiography. Results indicated that the percentage of S cells was similar in asbestos-treated and untreated cultures. Stimulation of the S phase also seems unlikely because treatment of RPMCs with asbestos fibres in the absence of HU resulted in a reduction of [3H]dThd incorporation attributed to an impairment of the S phase by the fibres. 1-4 micrograms/ml benzo[a]pyrene or 10-50 J/m2 UV light resulted in an approximate doubling of [3H]dThd incorporation. The effects of inhibitors of DNA repair were determined in chrysotile-treated RPMCs. [3H]dThd incorporation was inhibited by cytosine arabinoside and nalidixic acid. These results show that asbestos produces UDS in RPMCs.

  2. A novel {sup 125}I-labeled daunorubicin derivative for radionuclide-based cancer therapy

    Energy Technology Data Exchange (ETDEWEB)

    Ickenstein, Ludger M. [Department of Physical and Analytical Chemistry, Uppsala University, P.O. Box 579, 75121 Uppsala (Sweden); Edwards, Katarina [Department of Physical and Analytical Chemistry, Uppsala University, P.O. Box 579, 75121 Uppsala (Sweden); Sjoeberg, Stefan [Department of Biochemistry and Organic Chemistry, Uppsala University, P.O. Box 576, 75123 Uppsala (Sweden); Carlsson, Joergen [Rudbeck Laboratory, Division of Biomedical Radiation Sciences, Department of Oncology, Radiology and Clinical Immunology, Uppsala University, 75185 Uppsala (Sweden); Gedda, Lars [Rudbeck Laboratory, Division of Biomedical Radiation Sciences, Department of Oncology, Radiology and Clinical Immunology, Uppsala University, 75185 Uppsala (Sweden) and Rudbeck Laboratory, Division of Experimental Urology, Department of Surgery, Uppsala University, 75185 Uppsala (Sweden)]. E-mail:


    Introduction: Auger electron emitters, such as {sup 125}I, are getting increasingly wider recognition as alternatives to current anticancer treatments. The effectiveness of Auger electrons is strongly dependent on their proximity to DNA and is therefore considered as harmless outside the nucleus. Methods: {sup 125}I or {sup 127}I was conjugated with Comp1, Comp2 or Comp3 - three derivatives of the chemotherapeutic drug daunorubicin. Their capacity factors, DNA-binding constants and exclusion parameters, and the degree of DNA fragmentation after incubating isolated DNA with our {sup 127}I- or {sup 125}I-conjugated daunorubicin derivatives were determined. Human breast adenocarcinoma (SK-BR-3) cells were incubated with the derivatives; fluorescent microscopy and autoradiography images were generated; and cell growth was monitored. Results and Discussion: The capacity factor of {sup 127}I-Comp1 was similar to those of daunorubicin and doxorubicin, whereas lower capacity factors of {sup 127}I-Comp2 and {sup 127}I-Comp3 suggested reduced interactions with lipid membranes. DNA exclusion parameters and binding constants of {sup 127}I-Comp1 and {sup 127}I-Comp2, but not of {sup 127}I-Comp3, were similar to those of doxorubicin. Fluorescent microscopy and autoradiography images of SK-BR-3 cells revealed that {sup 127}I-Comp1 and {sup 125}I-Comp1 accumulated in tumor cell nuclei, whereas {sup 127}I-Comp2 and {sup 127}I-Comp3 were present predominantly in other cell compartments. The binding of {sup 125}I-Comp1 to isolated chromosomal DNA led to major fragmentation. Incubation of SK-BR-3 cells with {sup 125}I-Comp1 inhibited cell growth, whereas doxorubicin or {sup 127}I-Comp1 administered at the same concentration had no effect on cell growth. Our results thus suggest that {sup 125}I-Comp1 has the potential to become a new tool for anticancer therapy.

  3. Activated platelets in carotid artery thrombosis in mice can be selectively targeted with a radiolabeled single-chain antibody.

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    Timo Heidt

    Full Text Available BACKGROUND: Activated platelets can be found on the surface of inflamed, rupture-prone and ruptured plaques as well as in intravascular thrombosis. They are key players in thrombosis and atherosclerosis. In this study we describe the construction of a radiolabeled single-chain antibody targeting the LIBS-epitope of activated platelets to selectively depict platelet activation and wall-adherent non-occlusive thrombosis in a mouse model with nuclear imaging using in vitro and ex vivo autoradiography as well as small animal SPECT-CT for in vivo analysis. METHODOLOGY/PRINCIPAL FINDINGS: LIBS as well as an unspecific control single-chain antibody were labeled with (111Indium ((111In via bifunctional DTPA ( = (111In-LIBS/(111In-control. Autoradiography after incubation with (111In-LIBS on activated platelets in vitro (mean 3866 ± 28 DLU/mm(2, 4010 ± 630 DLU/mm(2 and 4520 ± 293 DLU/mm(2 produced a significantly higher ligand uptake compared to (111In-control (2101 ± 76 DLU/mm(2, 1181 ± 96 DLU/mm(2 and 1866 ± 246 DLU/mm(2 indicating a specific binding to activated platelets; P<0.05. Applying these findings to an ex vivo mouse model of carotid artery thrombosis revealed a significant increase in ligand uptake after injection of (111In-LIBS in the presence of small thrombi compared to the non-injured side, as confirmed by histology (49630 ± 10650 DLU/mm(2 vs. 17390 ± 7470 DLU/mm(2; P<0.05. These findings could also be reproduced in vivo. SPECT-CT analysis of the injured carotid artery with (111In-LIBS resulted in a significant increase of the target-to-background ratio compared to (111In-control (1.99 ± 0.36 vs. 1.1 ± 0.24; P < 0.01. CONCLUSIONS/SIGNIFICANCE: Nuclear imaging with (111In-LIBS allows the detection of platelet activation in vitro and ex vivo with high sensitivity. Using SPECT-CT, wall-adherent activated platelets in carotid arteries could be depicted in vivo. These results encourage further studies elucidating the role of

  4. Anti-CD20 Immunoglobulin G Radiolabeling with a 99mTc-Tricarbonyl Core: In Vitro and In Vivo Evaluations.

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    Hélène Carpenet

    Full Text Available In recent years, the diagnostic and therapeutic uses of radioisotopes have shown significant progress. Immunoglobulin (Ig appears to be a promising tracer, particularly due to its ability to target selected antigens. The main objective of this study is to optimize and assess an Ig radiolabeling method with Technetium 99m (99mTc, an attractive radioelement used widely for diagnostic imaging. Monoclonal anti-CD20 IgG was retained to study in vitro and in vivo radiolabeling impact. After IgG derivatization with 2-iminothiolane, IgG-SH was radiolabeled by an indirect method, using a 99mTc-tricarbonyl core. Radiolabeling stability was evaluated over 24h by thin-layer chromatography. IgG integrity was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis coupled with Western blot and autoradiography. The radiolabeled Ig's immunoaffinity was assessed in vitro by a radioimmunoassay method and binding experiments with cells (EL4-hCD20 and EL4-WT. Biodistribution studies were performed in normal BALB/c mice. Tumor uptake was assessed in mice bearing EL4-hCD20 and EL4-WT subcutaneous xenografts. With optimized method, high radiolabeling yields were obtained (95.9 ± 3.5%. 99mTc-IgG-SH was stable in phosphate-buffered saline (4°C and 25°C and in serum (37°C, even if important sensitivity to transchelation was observed. IgG was not degraded by derivatization and radiolabeling, as shown by Western blot and autoradiography results. 99mTc-anti-CD20 IgG-SH immunoaffinity was estimated with Kd = 35 nM by both methods. In vivo biodistribution studies for 48h showed significant accumulation of radioactivity in plasma, liver, spleen, lungs and kidneys. Planar scintigraphy of mice bearing tumors showed a significant uptake of 99mTc-anti-CD20 IgG-SH in CD20+ tumor versus CD20- tumor. Radiolabeling of derivatized IgG with 99mTc-tricarbonyl was effective, stable and required few antibody amounts. This attractive radiolabeling method is "antibody safe

  5. Radiosynthesis and biological evaluation of an {sup 18}F-labeled derivative of the novel pyrazolopyrimidine sedative-hypnotic agent indiplon

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    Hoepping, Alexander [ABX Advanced Biochemical Compounds GmbH, 01454 Radeberg (Germany); Scheunemann, Matthias [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany); Fischer, Steffen [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany); Deuther-Conrad, Winnie [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany); Hiller, Achim [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany); Wegner, Florian [Department of Neurology, University of Leipzig, 04103 Leipzig (Germany); Diekers, Michael [ABX Advanced Biochemical Compounds GmbH, 01454 Radeberg (Germany); Steinbach, Joerg [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany); Brust, Peter [Institute of Interdisciplinary Isotope Research, 04318 Leipzig (Germany)]. E-mail:


    Introduction: Gamma amino butyric acid type A (GABA{sub A}) receptors are involved in a variety of neurological and psychiatric diseases, which have promoted the development and use of radiotracers for positron emission tomography imaging. Radiolabeled benzodiazepine antagonists such as flumazenil have most extensively been used for this purpose so far. Recently, the non-benzodiazepine pyrazolopyrimidine derivative indiplon with higher specificity for the {alpha}{sub 1} subtype of the GABA{sub A} receptor has been introduced for treatment of insomnia. The aim of this study was the development and biological evaluation of an {sup 18}F-labeled derivative of indiplon. Methods: Both [{sup 18}F]fluoro-indiplon and its labeling precursor were synthesized by two-step procedures starting from indiplon. The radiosynthesis of [{sup 18}F]fluoro-indiplon was performed using the bromoacetyl precursor followed by multiple-stage purification using semipreparative HPLC and solid phase extraction. Stability, partition coefficients, binding affinities and regional brain binding were determined in vitro. Biodistribution and radiotracer metabolism were studied in vivo. Results: [{sup 18}F]Fluoro-indiplon was readily accessible in good yields (38-43%), with high purity and high specific radioactivity (>150 GBq/{mu}mol). It displays high in vitro stability and moderate lipophilicity. [{sup 18}F]Fluoro-indiplon has an affinity to GABA{sub A} receptors comparable to indiplon (K {sub i}=8.0 nM vs. 3.4 nM). In vitro autoradiography indicates high [{sup 18}F]fluoro-indiplon binding in regions with high densities of GABA{sub A} receptors. However, ex vivo autoradiography and organ distribution studies show no evidence of specific binding of [{sup 18}F]fluoro-indiplon. Furthermore, the radiotracer is rapidly metabolized with high accumulation of labeled metabolites in the brain. Conclusions: Although [{sup 18}F]fluoro-indiplon shows good in vitro features, it is not suitable for in vivo

  6. Distinct α subunit variations of the hypothalamic GABAA receptor triplets (αβγ are linked to hibernating state in hamsters

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    Alò Raffaella


    Full Text Available Abstract Background The structural arrangement of the γ-aminobutyric acid type A receptor (GABAAR is known to be crucial for the maintenance of cerebral-dependent homeostatic mechanisms during the promotion of highly adaptive neurophysiological events of the permissive hibernating rodent, i.e the Syrian golden hamster. In this study, in vitro quantitative autoradiography and in situ hybridization were assessed in major hypothalamic nuclei. Reverse Transcription Reaction-Polymerase chain reaction (RT-PCR tests were performed for specific GABAAR receptor subunit gene primers synthases of non-hibernating (NHIB and hibernating (HIB hamsters. Attempts were made to identify the type of αβγ subunit combinations operating during the switching ON/OFF of neuronal activities in some hypothalamic nuclei of hibernators. Results Both autoradiography and molecular analysis supplied distinct expression patterns of all α subunits considered as shown by a strong (p 1 ratio (over total α subunits considered in the present study in the medial preoptic area (MPOA and arcuate nucleus (Arc of NHIBs with respect to HIBs. At the same time α2 subunit levels proved to be typical of periventricular nucleus (Pe and Arc of HIB, while strong α4 expression levels were detected during awakening state in the key circadian hypothalamic station, i.e. the suprachiasmatic nucleus (Sch; 60%. Regarding the other two subunits (β and γ, elevated β3 and γ3 mRNAs levels mostly characterized MPOA of HIBs, while prevalently elevated expression concentrations of the same subunits were also typical of Sch, even though this time during the awakening state. In the case of Arc, notably elevated levels were obtained for β3 and γ2 during hibernating conditions. Conclusion We conclude that different αβγ subunits are operating as major elements either at the onset of torpor or during induction of the arousal state in the Syrian golden hamster. The identification of a brain regional

  7. Anatomical location of LPA1 activation and LPA phospholipid precursors in rodent and human brain. (United States)

    González de San Román, Estibaliz; Manuel, Iván; Giralt, María Teresa; Chun, Jerold; Estivill-Torrús, Guillermo; Rodríguez de Fonseca, Fernando; Santín, Luis Javier; Ferrer, Isidro; Rodríguez-Puertas, Rafael


    Lysophosphatidic acid (LPA) is a signaling molecule that binds to six known G protein-coupled receptors: LPA1 -LPA6 . LPA evokes several responses in the CNS, including cortical development and folding, growth of the axonal cone and its retraction process. Those cell processes involve survival, migration, adhesion proliferation, differentiation, and myelination. The anatomical localization of LPA1 is incompletely understood, particularly with regard to LPA binding. Therefore, we have used functional [(35) S]GTPγS autoradiography to verify the anatomical distribution of LPA1 binding sites in adult rodent and human brain. The greatest activity was observed in myelinated areas of the white matter such as corpus callosum, internal capsule and cerebellum. MaLPA1 -null mice (a variant of LPA1 -null) lack [(35) S]GTPγS basal binding in white matter areas, where the LPA1 receptor is expressed at high levels, suggesting a relevant role of the activity of this receptor in the most myelinated brain areas. In addition, phospholipid precursors of LPA were localized by MALDI-IMS in both rodent and human brain slices identifying numerous species of phosphatides and phosphatidylcholines. Both phosphatides and phosphatidylcholines species represent potential LPA precursors. The anatomical distribution of these precursors in rodent and human brain may indicate a metabolic relationship between LPA and LPA1 receptors. Lysophosphatidic acid (LPA) is a signaling molecule that binds to six known G protein-coupled receptors (GPCR), LPA1 to LPA6 . LPA evokes several responses in the central nervous system (CNS), including cortical development and folding, growth of the axonal cone and its retraction process. We used functional [(35) S]GTPγS autoradiography to verify the anatomical distribution of LPA1 -binding sites in adult rodent and human brain. The distribution of LPA1 receptors in rat, mouse and human brains show the highest activity in white matter myelinated areas. The basal and

  8. HPLC and TLC methods for analysis of [(18)F]FDG and its metabolites from biological samples. (United States)

    Rokka, Johanna; Grönroos, Tove J; Viljanen, Tapio; Solin, Olof; Haaparanta-Solin, Merja


    The most used positron emission tomography (PET) tracer, 2-[(18)F]fluoro-2-deoxy-d-glucose ([(18)F]FDG), is a glucose analogue that is used to measure tissue glucose consumption. Traditionally, the Sokoloff model is the basis for [(18)F]FDG modeling. According to this model, [(18)F]FDG is expected to be trapped in a cell in the form of [(18)F]FDG-6-phosphate ([(18)F]FDG-6-P). However, several studies have shown that in tissues, [(18)F]FDG metabolism goes beyond [(18)F]FDG-6-P. Our aim was to develop radioHPLC and radioTLC methods for analysis of [(18)F]FDG metabolites from tissue samples. The radioHPLC method uses a sensitive on-line scintillation detector to detect radioactivity, and the radioTLC method employs digital autoradiography to detect the radioactivity distribution on a TLC plate. The HPLC and TLC methods were developed using enzymatically in vitro-produced metabolites of [(18)F]FDG as reference standards. For this purpose, three [(18)F]FDG metabolites were synthesized: [(18)F]FDG-6-P, [(18)F]FD-PGL, and [(18)F]FDG-1,6-P2. The two methods were evaluated by analyzing the [(18)F]FDG metabolic profile from rodent ex vivo tissue homogenates. The HPLC method with an on-line scintillation detector had a wide linearity in a range of 5Bq-5kBq (LOD 46Bq, LOQ 139Bq) and a good resolution (Rs ≥1.9), and separated [(18)F]FDG and its metabolites clearly. The TLC method combined with digital autoradiography had a high sensitivity in a wide range of radioactivity (0.1Bq-2kBq, LOD 0.24Bq, LOQ 0.31Bq), and multiple samples could be analyzed simultaneously. As our test and the method validation with ex vivo samples showed, both methods are useful, and at best they complement each other in analysis of [(18)F]FDG and its radioactive metabolites from biological samples.

  9. Preclinical evaluation and test-retest studies of [{sup 18}F]PSS232, a novel radioligand for targeting metabotropic glutamate receptor 5 (mGlu{sub 5})

    Energy Technology Data Exchange (ETDEWEB)

    Milicevic Sephton, Selena; Mueller Herde, Adrienne; Keller, Claudia; Ruedisuehli, Sonja; Schibli, Roger; Kraemer, Stefanie D.; Ametamey, Simon M. [Center for Radiopharmaceutical Sciences of ETH, PSI and USZ, Zurich (Switzerland); Mu, Linjing [University Hospital Zuerich, Department of Nuclear Medicine, Zuerich (Switzerland); Auberson, Yves [Novartis Institutes for Biomedical Research, Novartis Pharma AG, Basel (Switzerland)


    A novel, {sup 18}F-labelled metabotropic glutamate receptor subtype 5 (mGlu{sub 5}) derivative of [{sup 11}C]ABP688 ([{sup 11}C]1), [{sup 18}F]PSS232 ([{sup 18}F]5), was evaluated in vitro and in vivo for its potential as a PET agent and was used in test-retest reliability studies The radiosynthesis of [{sup 18}F]5 was accomplished via a one-step reaction using a mesylate precursor. In vitro stability was determined in PBS and plasma, and with liver microsomal enzymes. Metabolite studies were performed using rat brain extracts, blood and urine. In vitro autoradiography was performed on horizontal slices of rat brain using 1 and 8, antagonists for mGlu{sub 5} and mGlu{sub 1}, respectively. Small-animal PET, biodistribution, and test-retest studies were performed in Wistar rats. In vivo, dose-dependent displacement studies were performed using 6 and blocking studies with 7. [{sup 18}F]5 was obtained in decay-corrected maximal radiochemical yield of 37 % with a specific activity of 80 - 400 GBq/μmol. Treatment with rat and human microsomal enzymes in vitro for 60 min resulted in 20 % and 4 % of hydrophilic radiometabolites, respectively. No hydrophilic decomposition products or radiometabolites were found in PBS or plasma. In vitro autoradiography on rat brain slices showed a heterogeneous distribution consistent with the known distribution of mGlu{sub 5} with high binding to hippocampal and cortical regions, and negligible radioactivity in the cerebellum. Similar distribution of radioactivity was found in PET images. Under displacement conditions with 6, reduced [{sup 18}F]5 binding was found in all brain regions except the cerebellum. 7 reduced binding in the striatum by 84 % on average. Test-retest studies were reproducible with a variability ranging from 6.8 % to 8.2 %. An extended single-dose toxicity study in Wistar rats showed no compound-related adverse effects. The new mGlu{sub 5} radiotracer, [{sup 18}F]5, showed specific and selective in vitro and in vivo

  10. Applications of scientific imaging in environmental toxicology (United States)

    El-Demerdash, Aref M.

    The national goals of clean air, clean water, and healthy ecosystems are a few of the primary forces that drive the need for better environmental monitoring. As we approach the end of the 1990s, the environmental questions at regional to global scales are being redefined and refined in the light of developments in environmental understanding and technological capability. Research in the use of scientific imaging data for the study of the environment is urgently needed in order to explore the possibilities of utilizing emerging new technologies. The objective of this research proposal is to demonstrate the usability of a wealth of new technology made available in the last decade to providing a better understanding of environmental problems. Research is focused in two imaging techniques macro and micro imaging. Several examples of applications of scientific imaging in research in the field of environmental toxicology were presented. This was achieved on two scales, micro and macro imaging. On the micro level four specific examples were covered. First, the effect of utilizing scanning electron microscopy as an imaging tool in enhancing taxa identification when studying diatoms was presented. Second, scanning electron microscopy combined with energy dispersive x-ray analyzer were demonstrated as a valuable and effective tool for identifying and analyzing household dust samples. Third, electronic autoradiography combined with FT-IR microscopy were used to study the distribution pattern of [14C]-Malathion in rats as a result of dermal exposure. The results of the autoradiography made on skin sections of the application site revealed the presence of [ 14C]-activity in the first region of the skin. These results were evidenced by FT-IR microscopy. The obtained results suggest that the penetration of Malathion into the skin and other tissues is vehicle and dose dependent. The results also suggest the use of FT-IR microscopy imaging for monitoring the disposition of

  11. {sup 18}F-Labelled metomidate analogues as adrenocortical imaging agents

    Energy Technology Data Exchange (ETDEWEB)

    Erlandsson, Maria; Karimi, Farhad [Department of Biochemistry and Organic Chemistry, Uppsala University, Box 576, S-751 23 Uppsala (Sweden); Lindhe, Orjan [Uppsala Imanet, GE Healthcare, Box 967, S-751 09 Uppsala (Sweden); Langstroem, Bengt [Department of Biochemistry and Organic Chemistry, Uppsala University, Box 576, S-751 23 Uppsala (Sweden)], E-mail:


    Introduction: Two- and one-step syntheses of {sup 18}F-labelled analogues of metomidate, such as 2-[{sup 18}F]fluoroethyl 1-[(1R)-1-phenylethyl]-1H-imidazole-5-carboxylate (1), 2-[{sup 18}F]fluoroethyl 1-[(1R)-1-(4-chlorophenyl)ethyl]-1H-imidazole-5-carboxylate (2), 2-[{sup 18}F]fluoroethyl 1-[(1R)-1-(4-bromophenyl)ethyl]-1H-imidazole-5-carboxylate (3), 3-[{sup 18}F]fluoropropyl 1-[(1R)-1-(4-bromophenyl)ethyl]-1H-imidazole-5-carboxylate (4) and 3-[{sup 18}F]fluoropropyl 1-[(1R)-1-phenylethyl]-1H-imidazole-5-carboxylate (5) are presented. Methods: Analogues 1-5 were prepared by a two-step reaction sequence that started with the synthesis of either 2-[{sup 18}F]fluoroethyl 4-methylbenzenesulfonate or 3-[{sup 18}F]fluoropropyl 4-methylbenzenesulfonate. These were used as {sup 18}F-alkylating agents in the second step, in which they reacted with the ammonium salt of a 1-[(1R)-1-phenylethyl]-1H-imidazole-5-carboxylic acid. One-step-labelling syntheses of 1, 2 and 5 were also explored. Analogues 1-4 were biologically validated by frozen-section autoradiography and organ distribution. Metabolite analysis was performed for 2 and 3. Results: The radiochemical yield of the two-step synthesis was in the range of 10-29% and that of the one-step synthesis was 25-37%. Using microwave irradiation in the one-step synthesis of 1 and 2 increased the radiochemical yield to 46{+-}3% and 79{+-}30%, respectively. Conclusion: Both the frozen-section autoradiography and organ distribution results indicated that analogue 2 has a potential as an adrenocortical imaging agent, having the highest degree of specific adrenal binding and best ratio of adrenal to organ uptake among the compounds studied.

  12. Effect of acute and chronic treatment with risperidone on the serotonin and dopamine receptors in the rat brain

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    Choi, Yun Young; Moon, Dae Hyuk; Son, Hye Kyung; Kim, Chang Yoon; Lee, Chul; Lee, Hee Kyung [College of Medicine, Ulsan Univ., Seoul (Korea, Republic of)


    The therapeutic efficacy of antipsychotic drugs is generally attributed to their ability to block dopamine D{sub 2} receptors. Classical D{sub 2} antagonists are not effective to treat negative symptoms and produce extrapyramidal side effects. On the other hand, atypical antipsychotic agents ameliorate negative symptoms without producing extrapyramidal side effects, and it is reported to be associated with blockade of serotonin 5-HT{sub 2} receptors. The purpose of this study was to evaluate the effect of risperidone on neuroreceptors in the rat brain by quantitative autoradiography method. In acute treatment group, risperidone was injected into peritoneal cavity of male Wistar rats with dose of 0, 0.1, 0.25, 0.5, 1.0 and 2.0mg/kg in each group (5/group), and they were decapitated after 2 hours. In chronic treatment group, risperidone was injected with dose of 0, 0.1, and 1m/kg (I.P.) for 21 ays and decapitated after 24 hours following last treatment. The effect of risperodone on the binding of [{sup 3}H) spiperone to 5-HT{sub 2} and D{sub 2} receptors were analysed in 4 discrete regions of the striatum, nucleus accumbens, and frontal cortex by quantitative autoradiography. Acute treatment with risperidone reduced cortical 5-HT{sub 2} specific [{sup 3}H]spiperone binding to 32% of vehicle-treated control. Subcortical 5-HR{sub 2} specific [{sup 3}H]spiperone binding was not affected at all dose groups whereas a significant reduction (57%) in D{sub 2} specific [{sup 3}H]spiperone binding was observed in risperidone treated group at doses of 1-2mg/kg. Chronic treatment with risperidone produced a decrease in the maximal number of cortical 5-HT{sub 2} receptors to 51% and 46% of control in 0.1mg/kg and 1mg/kg treated group respectively. In conclusion, risperidone is a cortical serotonin receptor antagonist with relatively weak antagonistic action on dopamine receptors. These effects on neuroreceptors may explain the therapeutic effect of risperidone as a atypical

  13. PET imaging of neuroinflammation in a rat traumatic brain injury model with radiolabeled TSPO ligand DPA-714

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Yu [Medical School of Southeast University, Jiangsu Key Laboratory of Molecular Imaging and Functional Imaging, Department of Radiology, Zhongda Hospital, Nanjing (China); National Institutes of Health - NIH, Laboratory of Molecular Imaging and Nanomedicine - LOMIN, National Institute of Biomedical Imaging and Bioengineering - NIBIB, Bethesda, MD (United States); Yue, Xuyi; Kiesewetter, Dale O.; Niu, Gang; Chen, Xiaoyuan [National Institutes of Health - NIH, Laboratory of Molecular Imaging and Nanomedicine - LOMIN, National Institute of Biomedical Imaging and Bioengineering - NIBIB, Bethesda, MD (United States); Teng, Gaojun [Medical School of Southeast University, Jiangsu Key Laboratory of Molecular Imaging and Functional Imaging, Department of Radiology, Zhongda Hospital, Nanjing (China)


    The inflammatory response in injured brain parenchyma after traumatic brain injury (TBI) is crucial in the pathological process. In order to follow microglia activation and neuroinflammation after TBI, we performed PET imaging in a rat model of TBI using {sup 18}F-labeled DPA-714, a ligand of the 18-kDa translocator protein (TSPO). TBI was induced in male SD rats by a controlled cortical impact. The success of the TBI model was confirmed by MRI. [{sup 18}F]DPA-714 was synthesized using a slightly modified TRACERLab FX-FN module and an automated procedure. In vivo PET imaging was performed at different time points after surgery using an Inveon small-animal PET scanner. The specificity of [{sup 18}F]DPA-714 was confirmed by a displacement study with an unlabeled competitive TSPO ligand, PK11195. Ex vivo autoradiography as well as immunofluorescence staining was carried out to confirm the in vivo PET results. Both in vivo T{sub 2}-weighted MR images and ex vivo TTC staining results revealed successful establishment of the TBI model. Compared with the sham-treated group, [{sup 18}F]DPA-714 uptake was significantly higher in the injured brain area on PET images. Increased lesion-to-normal ratios of [{sup 18}F]DPA-714 were observed in the brain of TBI rats on day 2 after surgery. Ratios peaked around day 6 (2.65 ± 0.36) and then decreased gradually to nearly normal levels on day 28. The displacement study using PK11195 confirmed the specific binding of [{sup 18}F]DPA-714 to TSPO. The results of ex vivo autoradiography were consistent with in vivo PET results. Immunofluorescence staining showed the time course of TSPO expression after TBI and the temporal and the spatial distribution of microglia in the damaged brain area. TSPO-targeted PET using [{sup 18}F]DPA-714 as the imaging probe can be used to dynamically monitor the inflammatory response after TBI in a noninvasive manner. This method will not only facilitate a better understanding of the inflammatory process

  14. Fiber-optic system for dual-modality imaging of glucose probes 18F-FDG and 6-NBDG in atherosclerotic plaques.

    Directory of Open Access Journals (Sweden)

    Raiyan T Zaman

    Full Text Available Atherosclerosis is a progressive inflammatory condition that underlies coronary artery disease (CAD-the leading cause of death in the United States. Thus, the ultimate goal of this research is to advance our understanding of human CAD by improving the characterization of metabolically active vulnerable plaques within the coronary arteries using a novel catheter-based imaging system. The aims of this study include (1 developing a novel fiber-optic imaging system with a scintillator to detect both 18F and fluorescent glucose probes, and (2 validating the system on ex vivo murine plaques.A novel design implements a flexible fiber-optic catheter consisting of both a radio-luminescence and a fluorescence imaging system to detect radionuclide 18F-fluorodeoxyglucose (18F-FDG and the fluorescent analog 6-(N-(7-Nitrobenz-2-oxa-1,3-diazol-4-ylamino-6-Deoxyglucose (6-NBDG, respectively. Murine macrophage-rich atherosclerotic carotid plaques were imaged ex vivo after intravenous delivery of 18F-FDG or 6-NBDG. Confirmatory optical imaging by IVIS-200 and autoradiography were also performed.Our fiber-optic imaging system successfully visualized both 18F-FDG and 6-NBDG probes in atherosclerotic plaques. For 18F-FDG, the ligated left carotid arteries (LCs exhibited 4.9-fold higher radioluminescence signal intensity compared to the non-ligated right carotid arteries (RCs (2.6 × 10(4 ± 1.4 × 10(3 vs. 5.4 × 10(3 ± 1.3 × 10(3 A.U., P = 0.008. Similarly, for 6-NBDG, the ligated LCs emitted 4.3-fold brighter fluorescent signals than the control RCs (1.6 × 10(2 ± 2.7 × 10(1 vs. 3.8 × 10(1 ± 5.9 A.U., P = 0.002. The higher uptake of both 18F-FDG and 6-NBDG in ligated LCs were confirmed with the IVIS-200 system. Autoradiography further verified the higher uptake of 18F-FDG by the LCs.This novel fiber-optic imaging system was sensitive to both radionuclide and fluorescent glucose probes taken up by murine atherosclerotic plaques. In addition, 6-NBDG is a

  15. Radiosynthesis and evaluation of [{sup 11}C]YM-202074 as a PET ligand for imaging the metabotropic glutamate receptor type 1

    Energy Technology Data Exchange (ETDEWEB)

    Yanamoto, Kazuhiko; Konno, Fujiko; Odawara, Chika; Yamasaki, Tomoteru; Kawamura, Kazunori; Hatori, Akiko; Yui, Joji; Wakizaka, Hidekatsu [Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Nengaki, Nobuki [Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); SHI Accelerator Service Co., Ltd., Shinagawa-ku, Tokyo 141-8686 (Japan); Takei, Makoto [Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan); Tokyo Nuclear Service Co., Ltd., Taito-ku, Tokyo 110-0005 (Japan); Zhang Mingrong, E-mail: zhang@nirs.go.j [Molecular Imaging Center, National Institute of Radiological Sciences, Inage-ku, Chiba 263-8555 (Japan)


    Introduction: Developing positron emission tomography (PET) ligands for imaging metabotropic glutamate receptor type 1 (mGluR1) is important for studying its role in the central nervous system. N-cyclohexyl-6-{l_brace}[N-(2-methoxyethyl)-N-methylamino]methyl{r_brace} -N-methylthiazolo[3,2-a]benzimidazole-2-carboxamide (YM-202074) exhibited high binding affinity for mGluR1 (K{sub i}=4.8 nM), and selectivity over other mGluRs in vitro. The purpose of this study was to label YM-202074 with carbon-11 and to evaluate in vitro and in vivo characteristics of [{sup 11}C]YM-202074 as a PET ligand for mGluR1 in rodents. Methods: [{sup 11}C]YM-202074 was synthesized by N-[{sup 11}C]methylation of its desmethyl precursor with [{sup 11}C]methyl iodide. The in vitro and in vivo brain regional distributions were determined in rats using autoradiography and PET, respectively. Results: [{sup 11}C]YM-202074 (262-630 MBq, n=5) was obtained with radiochemical purity of >98% and specific activity of 27-52 GBq/{mu}mol at the end of synthesis, starting from [{sup 11}C]CO{sub 2} of 19.3-21.5 GBq. In vitro autoradiographic results showed that the high specific binding of [{sup 11}C]YM-202074 for mGluR1 was presented in the cerebellum, thalamus and hippocampus, which are known as mGluR1-rich regions. In ex vivo autoradiography and PET studies, the radioligand was specifically distributed in the cerebellum, although the uptake was low. Furthermore, the regional distribution was fairly uniform in the whole brain by pretreatment with JNJ16259685 (a mGluR1 antagonist). However, radiometabolite(s) was detected in the brain. Conclusions: From these results, especially considering the low brain uptake and the influx of radiometabolite(s) into brain, [{sup 11}C]YM-202074 may not be a useful PET ligand for in vivo imaging of mGluR1 in the brain.

  16. In Vivo Differences between Two Optical Isomers of Radioiodinated o-iodo-trans-decalinvesamicol for Use as a Radioligand for the Vesicular Acetylcholine Transporter.

    Directory of Open Access Journals (Sweden)

    Izumi Uno

    Full Text Available To develop a superior VAChT imaging probe for SPECT, radiolabeled (--OIDV and (+-OIDV were isolated and investigated for differences in their binding affinity and selectivity to VAChT, as well as their in vivo activities.Radioiodinated o-iodo-trans-decalinvesamicol ([125I]OIDV has a high binding affinity for vesicular acetylcholine transporter (VAChT both in vitro and in vivo. Racemic [125I]OIDV was separated into its two optical isomers (--[125I]OIDV and (+-[125I]OIDV by HPLC. To investigate VAChT binding affinity (Ki of two OIDV isomers, in vitro binding assays were performed. In vivo biodistribution study of each [125I]OIDV isomer in blood, brain regions and major organs of rats was performed at 2,30 and 60 min post-injection. In vivo blocking study were performed to reveal the binding selectivity of two [125I]OIDV isomers to VAChT in vivo. Ex vivo autoradiography were performed to reveal the regional brain distribution of two [125I]OIDV isomers and (--[123I]OIDV for SPECT at 60 min postinjection.VAChT binding affinity (Ki of (--[125I]OIDV and (+-[125I]OIDV was 22.1 nM and 79.0 nM, respectively. At 2 min post-injection, accumulation of (--[125I]OIDV was the same as that of (+-[125I]OIDV. However, (+-[125I]OIDV clearance from the brain was faster than (--[125I]OIDV. At 30 min post-injection, accumulation of (--[125I]OIDV (0.62 ± 0.10%ID/g was higher than (+-[125I]OIDV (0.46 ± 0.07%ID/g in the cortex. Inhibition of OIDV binding showed that (--[125I]OIDV was selectively accumulated in regions known to express VAChT in the rat brain, and ex vivo autoradiography further confirmed these results showing similar accumulation of (--[125I]OIDV in these regions. Furthermore, (--[123I]OIDV for SPECT showed the same regional brain distribution as (--[125I]OIDV.These results suggest that radioiodinated (--OIDV may be a potentially useful tool for studying presynaptic cholinergic neurons in the brain.

  17. Changes in postnatal norepinephrine alter alpha-2 adrenergic receptor development. (United States)

    Sanders, J D; Happe, H K; Bylund, D B; Murrin, L C


    Alpha-2 adrenergic receptors (A2AR) regulate multiple brain functions and are enriched in developing brain. Studies demonstrate norepinephrine (NE) plays a role in regulating brain maturation, suggesting it is important in A2AR development. To investigate this we employed models of NE absence and excess during brain development. For decreases in NE we used N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine hydrochloride (DSP4), a specific noradrenergic neurotoxin. Increased noradrenergic terminal density was produced by methylazoxymethanol acetate (MAM) treatment. A2AR density was assayed with [(3)H]RX821002 autoradiography. DSP4 lesions on postnatal day (PND) 3 produce A2AR decreases in many regions by PND 5. A2AR recover to control levels by PND 15 and 25 and there is no further change in total receptor density. We also assayed A2AR in brains lesioned with DSP4 on PND 13, 23, 33 and 43 and harvested 22 days post-lesion. A2AR levels remain similar to control at each of these time points. We examined A2AR functionality and high affinity state with epinephrine-stimulated [(35)S]GTPγS and [(125)I]p-iodoclonidine autoradiography, respectively. On PND 25, control animals and animals lesioned with DSP4 on PND 3 have similar levels of [(35)S]GTPγS incorporation and no change in high affinity state. This is in contrast to increases in A2AR high affinity state produced by DSP4 lesions of mature brain. We next investigated A2AR response to increases in norepinephrine levels produced by MAM. In contrast to DSP4 lesions, increasing NE results in a large increase in A2AR. Animals treated with MAM on gestational day 14 had cortical [(3)H]RX821002 binding 100-200% greater than controls on PND 25, 35, 45, 55 and 65. These data indicate that NE regulation of A2AR differs in developing and mature brain and support the idea that NE regulates A2AR development and this has long term effects on A2AR function.

  18. Characterization study of cesium concentrated particles in the soils near the Fukushima Daiichi nuclear power plant (United States)

    Satou, Yukihiko; Sueki, Keisuke; Sasa, Kimikazu; Adachi, Kouji; Igarashi, Yasuhito


    Radionuclides from the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident contaminated a vast area. Two types of contamination, spread and spot types, were observed in soils with autoradiography using an imaging plate. Other samples such as dust filters, vegetation, X-ray films, and so on, also indicate the spot type contamination in the early stage of the FDNPP accident. The source of spot type contamination is well known as hot particles at the Chernobyl Nuclear Power Plant (ChNPP) accident in 1986. Hot particles were divided into two groups, fuel hot particles and fission product particles, and they were emitted directly from reactor core with phreatic explosion and fire. In contrast, the official reports of the FDNPP accident did not conforme core explosion. In addition, the emitted total amount of Uranium was very few (Yamamoto et al., 2014). Thus, the spot type contaminations were not identified as the same of hot particles yet. Therefore, the present study aimed to pick up and identify the spot contaminations in soils. Surface soil samples were collected at 20 km northwest from the FDNPP in June 2013. Soils were spread in plastic bags for autoradiography with imaging plate analysis. Then, the soil particles were collected on a sticky carbon tape and analyzed by SEM-EDS to detect radioactive particles. Finally, particles were confirmed to contain photo peaks in the γ-spectrum by a germanium semiconductor detector. Four radioactive particles were isolated from the soil samples in the present study. Detected γ-ray emission radionuclides were only Cs-134 and Cs-137. The X-ray spectra on the SEM-EDS of all particles showed a Cs peak as well as O, Fe, Zn, and Rb peaks, and these elements were distributed uniformly within the particles. In addition, uniform distribution of Si was also shown. Moreover, U was detected from one of the particles, but U concentration was very low and existed locally in the particle. These characters are very similar to previous

  19. Quantitative immunolocalization of {mu} opioid receptors: regulation by naltrexone

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    Evans, C.J.; Lam, H.; To, T.; Anton, B. [Department of Psychiatry and Biobehavioral Sciences, Neuropsychiatric Institute, University of California, Los Angeles, CA (United States); Unterwald, E.M. [Department of Psychiatry, New York University Medical Center, New York, NY (United States)


    The present study utilized a newly developed quantitative immunohistochemical assay to measure changes in {mu} opioid receptor abundance following chronic administration of the opioid receptor antagonist naltrexone. These data were compared with those obtained from {mu} receptor radioligand binding on adjacent tissue sections, in order to determine whether the characteristic antagonist-induced increase in radioligand binding is due to an increase in the total number of {mu} receptors and/or to an increase in the proportion of receptors that are in an active binding conformation in the absence of a change in the total number of receptors. Adult male Sprague-Dawley rats were administered naltrexone, 7-8 mg/kg per day, or saline continuously for seven days by osmotic minipumps, after which time their brains were processed for immunohistochemistry and receptor autoradiography on adjacent fresh frozen tissue sections. Semiquantitative immunohistochemistry was performed using a radiolabelled secondary antibody for autoradiographic determination and a set of radioactive standards. Results demonstrate an overall concordance between the distribution of {mu} opioid receptors as measured by the two different methods with a few exceptions. Following naltrexone administration, {mu} receptor immunoreactivity was significantly higher in the amygdala, thalamus, hippocampus, and interpeduncular nucleus as compared with the saline-treated control animals. [{sup 3}H]D-Ala{sup 2},N-Me-Phe{sup 4},Gly-ol{sup 5}-enkephalin binding to {mu} opioid receptors was significantly higher in the globus pallidus, amygdala, thalamus, hypothalamus, hippocampus, substantia nigra, ventral tegmental area, central gray, and interpeduncular nucleus of the naltrexone-treated rats.These findings indicate that in some brain regions chronic naltrexone exposure increases the total number of {mu} opioid receptors, while in other regions there is an increase in the percent of active receptors without an

  20. Characterization of a novel rat cholangiocarcinoma cell culture model-CGCCA

    Institute of Scientific and Technical Information of China (English)

    Chun-Nan Yeh; Kun-Ju Lin; Tsung-Wen Chen; Ren-Ching Wu; Lee-Cheng Tsao; Ying-Tzu Chen; Wen-Hui Weng; Miin-Fu Chen


    AIM: To characterize a culture model of rat CCA cells, which were derived from a transplantable TTA-induced CCA and designated as Chang Gung CCA (CGCCA). METHODS: The CGCCA cells were cultured at in vitro passage 12 times on a culture dish in DMEM medium. To measure the doubling time, 103 cells were plated in a 96-well plate containing the growth medium. The cells were harvested 4 to 10 d after seeding, and a standard MTT assay was used to measure the growth. The phenotype of CACCA cell and xenograft was determined by immunohistochemical study. We also determine the chromosomal alterations of CGCCA, G-banding and spectral karyotyping studies were performed. The CGCCA cell line was transplanted into the nude mice for examining its tumorigenicity. 2-Deoxy-2-(18F)fluoro-Dglucose (FDG) autoradiography was also performed to evaluate the FDG uptake of the tumor xenograft. RESULTS: The doubling time for the CGCCA cell line was 32 h. After transplantation into nude mice, FDG autoradiography showed that the tumors formed at the cell transplantation site had a latency period of 4-6 wk with high FDG uptake excluding necrosis tissue. Moreover, immunohistochemical staining revealed prominent cytoplasmic expression of c-erb-B2, CK19, c-Met, COX-Ⅱ, EGFR, MUC4, and a negative expression of K-ras. All data confirmed the phenotypic features of the CGCCA cell line coincide with the xenograft mice tumors, indicating cells containing the tumorigenicity of CCA originated from CCA. In addition, karyotypic banding analysis showed that the diploid (2n) cell status combines with ring and giant rod marker chromosomes in these clones; either both types simultaneously appeared or only one type of marker chromosome in a pair appeared in a cell. The major materials contained in the marker chromosome were primarily identified from chromosome 4. CONCLUSION: The current CGCCA cell line may be used as a non-K-ras effect CCA model and to obtain information and reveal novel pathways for CCA. Further

  1. Detection of early stage atherosclerotic plaques using PET and CT fusion imaging targeting P-selectin in low density lipoprotein receptor-deficient mice

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Ikuko, E-mail: [RIKEN Center for Molecular Imaging Science, Kobe (Japan); Department of Cardiovascular Medicine, Saga University, Saga (Japan); Hasegawa, Koki [RIKEN Center for Molecular Imaging Science, Kobe (Japan); Department of Pathology and Experimental Medicine, Kumamoto University, Kumamoto (Japan); Wada, Yasuhiro [RIKEN Center for Molecular Imaging Science, Kobe (Japan); Hirase, Tetsuaki; Node, Koichi [Department of Cardiovascular Medicine, Saga University, Saga (Japan); Watanabe, Yasuyoshi, E-mail: [RIKEN Center for Molecular Imaging Science, Kobe (Japan)


    Highlights: ► P-selectin regulates leukocyte recruitment as an early stage event of atherogenesis. ► We developed an antibody-based molecular imaging probe targeting P-selectin for PET. ► This is the first report on successful PET imaging for delineation of P-selectin. ► P-selectin is a candidate target for atherosclerotic plaque imaging by clinical PET. -- Abstract: Background: Sensitive detection and qualitative analysis of atherosclerotic plaques are in high demand in cardiovascular clinical settings. The leukocyte–endothelial interaction mediated by an adhesion molecule P-selectin participates in arterial wall inflammation and atherosclerosis. Methods and results: A {sup 64}Cu-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid conjugated anti-P-selectin monoclonal antibody ({sup 64}Cu-DOTA-anti-P-selectin mAb) probe was prepared by conjugating an anti-P-selectin monoclonal antibody with DOTA followed by {sup 64}Cu labeling. Thirty-six hours prior to PET and CT fusion imaging, 3 MBq of {sup 64}Cu-DOTA-anti-P-selectin mAb was intravenously injected into low density lipoprotein receptor-deficient Ldlr-/- mice. After a 180 min PET scan, autoradiography and biodistribution of {sup 64}Cu-DOTA-anti-P-selectin monoclonal antibody was examined using excised aortas. In Ldlr-/- mice fed with a high cholesterol diet for promotion of atherosclerotic plaque development, PET and CT fusion imaging revealed selective and prominent accumulation of the probe in the aortic root. Autoradiography of aortas that demonstrated probe uptake into atherosclerotic plaques was confirmed by Oil red O staining for lipid droplets. In Ldlr-/- mice fed with a chow diet to develop mild atherosclerotic plaques, probe accumulation was barely detectable in the aortic root on PET and CT fusion imaging. Probe biodistribution in aortas was 6.6-fold higher in Ldlr-/- mice fed with a high cholesterol diet than in those fed with a normal chow diet. {sup 64}Cu-DOTA-anti-P-selectin m

  2. Diffusion in the uranium - plutonium system and self-diffusion of plutonium in epsilon phase; Diffusion dans le systeme uranium-plutonium et autodiffusion du plutonium epsilon

    Energy Technology Data Exchange (ETDEWEB)

    Dupuy, M. [Commissariat a l' Energie Atomique, Fontenay-Aux-Roses (France). Centre d' Etudes Nucleaires


    A survey of uranium-plutonium phase diagram leads to confirm anglo-saxon results about the plutonium solubility in {alpha} uranium (15 per cent at 565 C) and the uranium one in {zeta} phase (74 per cent at 565 C). Interdiffusion coefficients, for concentration lower than 15 per cent had been determined in a temperature range from 410 C to 640 C. They vary between 0.2 and 6 10{sup 12} cm{sup 2} s{sup -1}, and the activation energy between 13 and 20 kcal/mole. Grain boundary, diffusion of plutonium in a uranium had been pointed out by micrography, X-ray microanalysis and {alpha} autoradiography. Self-diffusion of plutonium in {epsilon} phase (bcc) obeys Arrhenius law: D = 2. 10{sup -2} exp -(18500)/RT. But this activation energy does not follow empirical laws generally accepted for other metals. It has analogies with 'anomalous' bcc metals ({beta}Zr, {beta}Ti, {beta}Hf, U{sub {gamma}}). (author) [French] Une etude du diagramme d'equilibre uranium-plutonium conduit a confirmer les resultats anglo-saxons relatifs a la solubilite du plutonium dans l'uranium {alpha} (15 pour cent a 565 C) et de l'uranium dans la phase {zeta} (74 pour cent a 565 C). Les coefficients de diffusion chimique, pour des concentrations inferieures a 15 pour cent ont ete determines a des temperatures comprises entre 410 et 640 C. Ils se situent entre 0.2 et 6. 10{sup 12} cm{sup 2} s{sup -1}. L'energie d'activation varie entre 13 et 20 kcal/mole. La diffusion intergranulaire du plutonium dans l'uranium a a ete mise en evidence par micrographie, microanalyse X et autoradiographie {alpha}. L' autodiffusion du plutonium {beta} cubique centree obeit a la loi d'Arrhenius D = 2. 10{sup -2} exp - (18500)/RT. Son energie d'activation n'obeit pas aux lois empiriques generalement admises pour les autres metaux. Elle possede des analogies avec les cubiques centres ''anormaux'' (Zr{beta}, Ti{beta}, Hf{beta}, U{gamma}). (auteur)

  3. The effects of X-ray radiation on mandibular bone of low-calcium diet rats

    Energy Technology Data Exchange (ETDEWEB)

    Kurita, Akihiko (Nippon Dental Univ., Tokyo (Japan))


    In an attempt to examine the effects of X-ray on osteoporosis, a single dose of 30 Gy was delivered to the mandible in rats given low-calcium diet. Serum levels of calcium (Ca) and inorganic phosphorus (P) were measured; and changes in bone salt were determined by autoradiography, microradiography, and roentgenography using an electron probe microanalyzer. Body weight was lower in the irradiated group than the non-irradiated group, irrespective of types of diet. The serum Ca levels in the irradiated group given a normal diet were significantly decreased on Days 3, 7, and 14 days after irradiation. When given a low-Ca diet, these levels tended to be lower in the irradiated group than the non-irradiated group on Day 7 or later. The serum levels of inorganic P were significantly lower in the irradiated group given a normal diet than the non-irradiated group on Day 3. Rats given a low-Ca diet had the same levels, irrespective of irradiation. Autoradiography revealed that Ca-45 retention in the whole jaw was slightly greater in the irradiated group than the non-irradiated group On Days 7 and 21. Rats given a low-Ca diet in both irradiated and non-irradiated groups had a greater Ca-45 retention than those given a normal diet. Microradiography revealed that bone formation-like changes, such as flat surface of the periodontal membrane at the intra-alveolar septum, were slightly noticeable in the irradiated group of rats given a normal diet on Day 21. Thinning of the intra-alveolar septum and decrease of the trabecula at the diaphysis were also noticeable in the irradiated group of rats given a low-Ca diet. Variation of X-ray intensity was more marked on Day 7 than on Day 21 in the irradiated group given a normal diet. When given a low-Ca diet, both the irradiated and non-irradiated group had noticeable X-ray intensity variation. (N.K.).

  4. Whole-body biodistribution and brain PET imaging with [{sup 18}F]AV-45, a novel amyloid imaging agent - a pilot study

    Energy Technology Data Exchange (ETDEWEB)

    Lin, K.-J. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taiwan (China); Hsu, W.-C. [Department of Neurology, Chang Gung Memorial Hospital, Taiwan (China); Hsiao, I.-T.; Wey, S.-P. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taiwan (China); Jin, L.-W. [M.I.N.D. Institute and Department of Pathology, University of California, Davis, CA (United States); Skovronsky, Daniel [Avid Radiopharmaceuticals, Inc., Philadelphia, PA (United States); Wai, Y.-Y. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Radiology, Chang Gung Memorial Hospital, Taiwan (China); Chang, H.-P.; Lo, C.-W.; Yao, C.H.; Yen, T.-C. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Kung, M.-P. [Department of Nuclear Medicine and Molecular Imaging Center, Chang Gung Memorial Hospital, Taiwan (China); Department of Radiology, University of Pennsylvania, Philadelphia, PA (United States)


    Purpose: The compound (E)-4-(2-(6-(2-(2-(2-{sup 18}F-fluoroethoxy)ethoxy)ethoxy) pyridin-3-yl)vinyl)-N-methylbenzenamine ([{sup 18}F]AV-45) is a novel radiopharmaceutical capable of selectively binding to {beta}-amyloid (A{beta}) plaques. This pilot study reports the safety, biodistribution, and radiation dosimetry of [{sup 18}F]AV-45 in human subjects. Methods: In vitro autoradiography and fluorescent staining of postmortem brain tissue from patients with Alzheimer's disease (AD) and cognitively healthy subjects were performed to assess the specificity of the tracer. Biodistribution was assessed in three healthy elderly subjects (mean age: 60.0{+-}5.2 years) who underwent 3-h whole-body positron emission tomography (PET)/computed tomographic (CT) scans after a bolus injection of 381.9{+-}13.9 MBq of [{sup 18}F]AV-45. Another six subjects (three AD patients and three healthy controls, mean age: 67.7{+-}13.6 years) underwent brain PET studies. Source organs were delineated on PET/CT. All subjects underwent magnetic resonance imaging (MRI) for obtaining structural information. Results: In vitro autoradiography revealed exquisitely high specific binding of [{sup 18}F]AV-45 to postmortem AD brain sections, but not to the control sections. There were no serious adverse events throughout the study period. The peak uptake of the tracer in the brain was 5.12{+-}0.41% of the injected dose. The highest absorbed organ dose was to the gallbladder wall (184.7{+-}78.6 {mu}Gy/MBq, 4.8 h voiding interval). The effective dose equivalent and effective dose values for [{sup 18}F]AV-45 were 33.8{+-}3.4 {mu}Sv/MBq and 19.3{+-}1.3 {mu}Sv/MBq, respectively. Conclusion: [{sup 18}F]AV-45 binds specifically to A{beta} in vitro, and is a safe PET tracer for studying A{beta} distribution in human brain. The dosimetry is suitable for clinical and research application.

  5. Bone marrow uptake of {sup 99m}Tc-MIBI in patients with multiple myeloma

    Energy Technology Data Exchange (ETDEWEB)

    Fonti, R. [Medicina Nucleare, Facolta di Medicina, Univ. Federico II, Naples (Italy); Centro per la Medicina Nucleare C.N.R., Cattedra di Medicina Nucleare, Napoli (Italy); Del Vecchio, S.; Zannetti, A.; Di Gennaro, F.; Pace, L.; Salvatore, M. [Centro per la Medicina Nucleare C.N.R., Cattedra di Medicina Nucleare, Napoli (Italy); De Renzo, A.; Catalano, L.; Califano, C.; Rotoli, B. [Cattedra di Ematologia, Dipt. di Medicina Clinica e Sperimentale, Napoli (Italy)


    In a previous study, we showed the ability of technetium-99m methoxyisobutylisonitrile ({sup 99m}Tc-MIBI) scan to identify active disease in patients with multiple myeloma (Eur J Nucl Med 1998; 25: 714-720). In particular, a semiquantitative score of the extension and intensity of bone marrow uptake was derived and correlated with both the clinical status of the disease and plasma cell bone marrow infiltration. In order to estimate quantitatively {sup 99m}Tc-MIBI bone marrow uptake and to verify the intracellular localization of the tracer, bone marrow samples obtained from 24 multiple myeloma patients, three patients with monoclonal gammopathy of undetermined significance (MGUS) and two healthy donors were studied for in vitro uptake. After centrifugation over Ficoll-Hypaque gradient, cell suspensions were incubated with {sup 99m}Tc-MIBI and the uptake was expressed as the percentage of radioactivity specifically retained within the cells. The cellular localization of the tracer was assessed by micro-autoradiography. Twenty-two out of 27 patients underwent {sup 99m}Tc-MIBI scan within a week of bone marrow sampling. Whole-body images were obtained 10 min after intravenous injection of 555 MBq of the tracer; the extension and intensity of {sup 99m}Tc-MIBI uptake were graded using the semiquantitative score. A statistically significant correlation was found between in vitro uptake of {sup 99m}Tc-MIBI and both plasma cell infiltration (Pearson's coefficient of correlation r=0.69, P<0.0001) and in vivo score (Spearman rank correlation coefficient r=0.60, P<0.01). No specific tracer uptake was found in bone marrow samples obtained from the two healthy donors. Micro-autoradiography showed localization of {sup 99m}Tc-MIBI inside the plasma cells infiltrating the bone marrow. Therefore, our findings show that the degree of tracer uptake both in vitro and in vivo is related to the percentage of infiltrating plasma cells which accumulate the tracer in their inner

  6. A comprehensive study on the putative δ-opioid receptor (sub)types using the highly selective δ-antagonist, Tyr-Tic-(2S,3R)-β-MePhe-Phe-OH. (United States)

    Birkas, Erika; Bakota, Lidia; Gulya, Karoly; Wen, Ting; Pintar, John; Tóth, Geza; Szucs, Maria


    The goal of our work was a throughout characterization of the pharmacology of the TIPP-analog, Tyr-Tic-(2S,3R)-β-MePhe-Phe-OH and see if putative δ-opioid receptor subtypes can be distinguished. Analgesic latencies were assessed in mouse tail-flick assays after intrathecal administration. In vitro receptor autoradiography, binding and ligand-stimulated [(35)S]GTPγS functional assays were performed in the presence of putative δ(1)-(DPDPE: agonist, BNTX: antagonist), δ(2)-(agonist: deltorphin II, Ile(5,6)-deltorphin II, antagonist: naltriben) and μ-(DAMGO: agonist) opioid ligands. The examined antagonist inhibited the effect of DPDPE by 60%, but did not antagonize δ(2)- and μ-agonist induced analgesia. The radiolabeled form identified binding sites with K(D)=0.18 nM and receptor densities of 102.7 fmol/mg protein in mouse brain membranes. The binding site distribution of the [(3)H]Tyr-Tic-(2S,3R)-β-MePhe-Phe-OH agreed well with that of [(3)H]Ile(5,6)-deltorphin II as revealed by receptor autoradiography. Tyr-Tic-(2S,3R)-β-MePhe-Phe-OH displayed 2.49±0.06 and 0.30±0.01 nM potency against DPDPE and deltorphin II in the [(35)S]GTPγS functional assay, respectively. The rank order of potency of putative δ(1)- and δ(2)-antagonists against DPDPE and deltorphin was similar in brain and CHO cells expressing human δ-opioid receptors. Deletion of the DOR-1 gene resulted in no residual binding of the radioligand and no significant DPDPE effect on G-protein activation. Tyr-Tic-(2S,3R)-β-MePhe-Phe-OH is a highly potent and δ-opioid specific antagonist both in vivo and in vitro. However, the putative δ(1)- and δ(2)-opioid receptors could not be unequivocally distinguished in vitro.

  7. {sup 99m}Tc-Hynic-annexin V imaging to evaluate inflammation and apoptosis in rats with autoimmune myocarditis

    Energy Technology Data Exchange (ETDEWEB)

    Tokita, Naoki; Izumi, Tohru [Department of Internal Medicine, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara-city, Kanagawa 228-8555 (Japan); Hasegawa, Shinji; Maruyama, Kaoru [Division of Tracer Kinetics, Biomedical Research Center, Osaka University Graduate School of Medicine, Suita, Osaka (Japan); Blankenberg, Francis G. [Division of Pediatric Radiology, Stanford University School of Medicine, Lucile Salter Packard Children' s Hospital, Stanford, Calif. (United States); Tait, Jonathan F. [Department of Laboratory Medicine, University of Washington, Seattle, Wash. (United States); Strauss, William H. [Department of Nuclear Medicine, Memorial Sloan-Kettering Cancer Center, New York, New York (United States); Nishimura, Tsunehiko [Department of Radiology, Kyoto Prefectural University of Medicine, Kyoto, Kyoto (Japan)


    Inflammation and cell death are two important components of myocarditis. We evaluated the distribution of inflammation and apoptotic cell death in rats with autoimmune myocarditis using two radiotracers - technetium-99m Hynic-annexin V ({sup 99m}Tc-annexin) as a marker of apoptotic cell death and carbon-14 deoxyglucose ({sup 14}C-DG) as a marker of inflammation - in comparison with histologic findings. Three, 7 and 14 weeks after immunization with porcine cardiac myosin (acute, subacute, and chronic phases, respectively) {sup 99m}Tc-annexin and {sup 14}C-DG were injected. The uptake in the total heart was determined as the percentage of injected dose per gram (% ID/g) by tissue counting. Dual-tracer autoradiography with {sup 99m}Tc-annexin and {sup 14}C-DG was performed. The distribution of each of these agents was compared with the results of hematoxylin and eosin staining to identify areas of inflammation, and TUNEL staining to identify areas of apoptosis. Total cardiac uptake of {sup 99m}Tc-annexin in the acute phase of myocarditis was significantly higher than that in normal rats (1.28%{+-}0.30% vs 0.46%{+-}0.01%; P<0.0001); it then decreased in the subacute phase and reached normal levels (0.56%{+-}0.08% vs 0.60%{+-}0.08%; P=NS). Total cardiac uptake of {sup 14}C-DG in the acute phase of myocarditis was significantly higher than that in normal rats (2.78%{+-}0.95% vs 1.02%{+-}0.25%; P<0.0001); it then decreased in the subacute phase, but still remained higher than in controls (2.06%{+-}0.52% vs 1.37%{+-}0.46%; P<0.05). Using autoradiography and staining of tissue specimens, it was found that most histologic inflammatory foci corresponded to areas of high {sup 14}C-DG uptake; some also corresponded to areas of high {sup 99m}Tc-annexin uptake in the acute phase of myocarditis. {sup 99m}Tc-annexin localization was strongly correlated with the number of TUNEL-positive cells (P<0.0001, r=0.83), but the uptake of {sup 14}C-DG showed no relationship with it. There is

  8. Automated Detection of Trinucleotide Repeats in Fragile X Syndrome. (United States)

    Hamdan; Tynan; Fenwick; Leon


    Background: The conventional method for diagnosis of fragile X syndrome has been amplification of the trinucleotide repeat region of the FMR-1 gene by polymerase chain reaction (PCR) and Southern blot analysis to detect full expansion and hypermethylation. "Stuttering" resulting from incomplete amplification is still observed in the PCR products despite the use of reagents that reduce the secondary structure of the GC-rich template. In addition, PCR products can be detected by autoradiography only after 1 to 2 days of exposure. By combination of a recently reported amplification protocol with fluorescence detection of PCR products in an automated DNA sequencer, the PCR protocol for amplification of trinucleotide repeats was simplified. This modified protocol is highly reproducible, more accurate, and less costly than the conventional protocol because of the elimination of radioisotopes from the PCR. Methods and Results: PCRs were conducted with betaine and Pfu DNA polymerase. This improved PCR protocol allowed immediate detection of PCR products in agarose gels containing ethidium bromide. Stuttering was completely eliminated and fragments of up to 1kb ( approximately 250 repeats) were visible in agarose gels. PCR products were automatically detected by laser fluorescence in an automated DNA sequencer by inclusion of a fluorescently-labeled primer in the PCR reaction. A short electrophoresis run of 100 minutes in denaturing acrylamide gels was sufficient to give high resolution of fragments with higher accuracy and sensitivity than conventional detection by autoradiography. Conclusions: A simple, nonradioactive protocol that is more rapid and less expensive than the conventional PCR protocol for the detection of trinucleotide repeats has been developed. By use of this detection protocol, fragment sizes containing up to 100 repeats could be detected, alleles differing by one trinucleotide repeat were clearly resolved, and heterogeneous repeat patterns such as those

  9. Identification of macrophage external membrane proteins and their possible role in cell adhesion. (United States)

    Pearlstein, E; Dienstman, S R; Defendi, V


    Starch-activated mouse peritoneal macrophages (STpMAC) plated on plastic demonstrate the adhesive properties typical for activated pMAC: attaching as round cells and, within 15 min, spreading out with marginal membrane ruffles. These attached STpMAC were labeled by lactoperoxidase-catalysed 125I surface iodination, sodium dodecyl-sulfate-lysed, and the lysates electrophoresed on polyacrylamide gels which were examined by autoradiography. The STpMAC morphological phenotype correlates with the labeling of a particular protein (195,000, estimated mol wt). Normal pMAC (NpMAC), from unstimulated mice, do not spread and do not display the 195,000 band. Both pMAC band patterns, including the 195,000 band, are relatively resistant to trypsin digestion, as is pMAC adhesion itself trypsin-resistant. Neither class of pMAC exhibits fibronectin (Cell Adhesion Factor, LETS protein) which is a component in the adhesive matrix of cells forming trypsin-sensitive monolayers. When pMAC are tested against antifibronectin antibody, these cells do not give immunofluorescent staining. In summary, two functions in pMAC adhesion, enzyme resistance and the ability to spread, appear related to molecular properties distinctive for pMAC surface protein.

  10. Tyrosine Sulfation of Statherin

    Directory of Open Access Journals (Sweden)

    C. Kasinathan, N. Gandhi, P. Ramaprasad, P. Sundaram, N. Ramasubbu


    Full Text Available Tyrosylprotein sulfotransferase (TPST, responsible for the sulfation of a variety of secretory and membrane proteins, has been identified and characterized in submandibular salivary glands (William et al. Arch Biochem Biophys 1997; 338: 90-96. In the present study we demonstrate the sulfation of a salivary secretory protein, statherin, by the tyrosylprotein sulfotransferase present in human saliva. Optimum statherin sulfation was observed at pH 6.5 and at 20 mm MnCl2. Increase in the level of total sulfation was observed with increasing statherin concentration. The Km value of tyrosylprotein sulfotransferase for statherin was 40 μM. Analysis of the sulfated statherin product on SDS-polyacrylamide gel electrophoresis followed by autoradiography revealed 35S-labelling of a 5 kDa statherin. Further analysis of the sulfated statherin revealed the sulfation on tyrosyl residue. This study is the first report demonstrating tyrosine sulfation of a salivary secretory protein. The implications of this sulfation of statherin in hydroxyapatite binding and Actinomyces viscosus interactions are discussed.

  11. Penetration of ASM 981 in canine skin: a comparative study. (United States)

    Gutzwiller, Meret E Ricklin; Reist, Martin; Persohn, Elke; Peel, John E; Roosje, Petra J


    ASM 981 has been developed for topical treatment of inflammatory skin diseases. It specifically inhibits the production and release of pro-inflammatory cytokines. We measured the skin penetration of ASM 981 in canine skin and compared penetration in living and frozen skin. To make penetration of ASM 981 visible in dog skin, tritium labelled ASM 981 was applied to a living dog and to defrosted skin of the same dog. Using qualitative autoradiography the radioactive molecules were detected in the lumen of the hair follicles until the infundibulum, around the superficial parts of the hair follicles and into a depth of the dermis of 200 to 500 microm. Activity could not be found in deeper parts of the hair follicles, the dermis or in the sebaceous glands. Penetration of ASM 981 is low in canine skin and is only equally spread in the upper third of the dermis 24 hours after application. Penetration in frozen skin takes even longer than in living canine skin but shows the same distribution.

  12. Receptor-mediated binding and uptake of GnRH agonist and antagonist by pituitary cells

    Energy Technology Data Exchange (ETDEWEB)

    Jennes, L.; Stumpf, W.E.; Conn, P.M.


    The intracellular pathway of an enzyme resistant GnRH agonist (D- Lys6 -GnRH) conjugated to ferritin or to colloidal gold was followed in cultured pituitary cells. After an initial uniform distribution over the cell surface of gonadotropes, the electrondense marker was internalized, either individually or in small groups. After longer incubation times, the marker appeared in the lysosomal compartment and the Golgi apparatus, where it could be found in the vesicular as well as cisternal portion. In addition, the receptor-mediated endocytosis of the GnRH antagonist D-p-Glu1-D-Phe2-D-Trp3-D- Lys6 -GnRH was studied by light and electron microscopic autoradiography after 30 and 60 min of incubation to ensure uptake. At both time points, in in vitro as well as in vivo studies, silver grains were localized over cytoplasmic organelles of castration cells, including dilated endoplasmic reticulum, lysosomes, and clear vesicles. No consistent association with cell nuclei, mitochondria, or secretory vesicles could be observed. The results suggest that both agonist and antagonist are binding selectively to the plasma membrane of gonadotropes and subsequently are taken up via receptor-mediated endocytosis for degradation or possible action on synthetic processes.

  13. Pharmacokinetics, distribution, metabolism, and excretion of the dual reuptake inhibitor [(14)C]-nefopam in rats. (United States)

    Yu, Jian; Solon, Eric; Shen, Helen; Modi, Nishit B; Mittur, Aravind


    1. This study examined the pharmacokinetics, distribution, metabolism, and excretion of [(14)C] nefopam in rats after a single oral administration. Blood, plasma, and excreta were analyzed for total radioactivity, nefopam, and metabolites. Metabolites were profiled and identified. Radioactivity distribution was determined by quantitative whole-body autoradiography. 2. The pharmacokinetic profiles of total radioactivity and nefopam were similar in male and female rats. Radioactivity partitioned approximately equally between plasma and red blood cells. A majority of the radioactivity was excreted in urine within 24 hours and mass balance was achieved within 7 days. 3. Intact nefopam was a minor component in plasma and excreta. Numerous metabolites were identified in plasma and urine generated by multiple pathways including: hydroxylation/oxidation metabolites (M11, M22a and M22b, M16, M20), some of which were further glucuronidated (M6a to M6c, M7a to M7c, M8a and M8b, M3a to M3d); N-demethylation of nefopam to metabolite M21, which additionally undergoes single or multiple hydroxylations or sulfation (M9, M14, M23), with some of the hydroxylated metabolites further glucuronidated (M2a to M2d). 4. Total radioactivity rapidly distributed with highest concentrations found in the urinary bladder, stomach, liver, kidney medulla, small intestine, uveal tract, and kidney cortex without significant accumulation or persistence. Radioactivity reversibly associated with melanin-containing tissues.

  14. Colloid Facilitated Transport of Plutonium in Fractured Volcanic Tuff (United States)

    Kersting, A. B.; Zhao, P.; Walensky, J. R.; Roberts, S. K.; Johnson, M. R.; Zavarin, M.; Ramon, E. C.


    The transport of low-solubility radionuclides in a colloidal- or colloidal bound state is frequently suspected or observed. Groundwater contaminated with radionuclides associated with underground nuclear tests was collected from several different well locations at the Nevada Test Site (NTS). In each case, the low-levels of plutonium detected in the groundwater were overwhelmingly (>95percent) associated with the colloidal and not the dissolved fraction of the groundwater. The colloidal fractions consisted of secondary minerals such as clays and zeolites. To better understand the mechanisms controlling the potential colloidal transport of plutonium, colloid-facilitated fracture flow laboratory experiments are being conducted. Pseudocolloids consisting of Pu(IV) sorbed to clinoptilolite were combined with a radionuclide solution cocktail consisting of Np, U, Cs, Sr, Sm and 3H and Re (analog to Tc) tracers in NTS-type synthetic groundwater (4.5mM NaHCO3-). The cocktail was injected into a smooth fracture in a volcanic tuff rock core from the NTS and the effluent analyzed. Autoradiography and secondary ion mass spectrometry will be used to understand the mineral -colloid-radionuclide interactions in the fracture volcanic tuff.

  15. Effects of juvenile isolation and morphine treatment on social interactions and opioid receptors in adult rats: behavioural and autoradiographic studies. (United States)

    Van den Berg, C L; Van Ree, J M; Spruijt, B M; Kitchen, I


    The consequences of juvenile isolation and morphine treatment during the isolation period on (social) behaviour and mu-, delta- and kappa-opioid receptors in adulthood were investigated by using a social interaction test and in vitro autoradiography in rats. Juvenile isolation reduced social exploration in adults. Morphine treatment counteracted this reduction in isolated rats, but decreased social exploration in nonisolated rats. Self-grooming and nonsocial exploration were enhanced after juvenile isolation. Morphine treatment had no effect on self-grooming, but suppressed nonsocial exploration in isolated rats. With respect to the opioid receptors, juvenile isolation resulted in regiospecific increases in mu-binding sites with a 58% increase in the basolateral amygdala and a 33% increase in the bed nucleus of stria terminalis. Morphine treatment in isolated rats reversed this upregulation in both areas. The number of delta-binding sites did not differ between the experimental groups. A general upregulation of kappa-binding sites was observed after juvenile isolation, predominantly in the cortical regions, the hippocampus and the substantia nigra. Morphine treatment did not affect the upregulation of kappa-receptors. The results show that juvenile isolation during the play period causes long-term effects on social and nonsocial behaviours and on the number of mu- and kappa- but not delta-opioid receptors in distinct brain areas. The number of mu-receptors in the basolateral amygdala appears to be negatively correlated with the amount of social exploration in adult rats.

  16. Morphine treatment during juvenile isolation increases social activity and opioid peptides release in the adult rat. (United States)

    Van den Berg, C L; Kitchen, I; Gerrits, M A; Spruijt, B M; Van Ree, J M


    The consequences of juvenile isolation and morphine treatment on general activity, social activity and endogenous opioid release during a social interaction test were investigated in the adult rat. Rats were either isolated or socially housed during weeks 4 and 5 of age and treated daily during this isolation period subcutaneously with either saline or morphine. Directly after a social interaction test at 10 weeks of age, rats were injected with [3H]-diprenorphine and subsequently prepared for in vivo autoradiography. The autoradiographic technique was used to visualise neuroanatomical changes in opioid receptor occupancy, probably reflecting changes in opioid peptide release, as a result of social activity. Juvenile isolation increased general activity during the social interaction test, an effect which was accompanied by a reduction of opioid receptor occupancy in many brain areas, suggesting an increased opioid peptide release as a consequence of socially-induced general activity. Morphine treatment in isolated rats caused an increase in adult social activity and enhanced opioid peptide release in some cortical regions and the ventral tegmental area as compared to saline treated rats. Both social activity and opioid receptor occupancy were unaffected by morphine treatment in non-isolated rats. The present study underscores the role of opioid systems in adult social behaviors as a consequence of juvenile isolation. The results suggest a relationship between social activity and opioid peptide release during social contact. Increased social activity seems to be accompanied by elevated opioid peptide release in distinct brain areas after morphine treatment during juvenile isolation.

  17. Up-regulation of -opioid receptors in the spinal cord of morphine-tolerant rats

    Indian Academy of Sciences (India)

    Subrata Basu Ray; Himanshu Gupta; Yogendra Kumar Gupta


    Though morphine remains the most powerful drug for treating pain, its effectiveness is limited by the development of tolerance and dependence. The mechanism underlying development of tolerance to morphine is still poorly understood. One of the factors could be an alteration in the number of m-receptors within specific parts of the nervous system. However, reports on changes in the -opioid receptor density in the spinal cord after chronic morphine administration are conflicting. Most of the studies have used subcutaneously implanted morphine pellets to produce tolerance. However, it does not simulate clinical conditions, where it is more common to administer morphine at intervals, either by injections or orally. In the present study, rats were made tolerant to morphine by injecting increasing doses of morphine (10–50 mg/kg, subcutaneously) for five days. In vitro tissue autoradiography for localization of -receptor in the spinal cord was done using [3H]-DAMGO. As compared to the spinal cord of control rats, the spinal cord of tolerant rats showed an 18.8% increase or up-regulation in the density of -receptors in the superficial layers of the dorsal horn. This up-regulation of -receptors after morphine tolerance suggests that a fraction of the receptors have been rendered desensitized, which in turn could lead to tolerance.

  18. Medical application of radioisotopes

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Chang Woon; Lim, S. M.; Kim, E. H. [and others


    In this project, we studied following subjects: 1. Clinical research for radionuclide therapy 2. Development of in vitro assay method with radioisotope 3. Development of binary therapy; Boron neutron capture therapy and photodynamic therapy 4. Development of diagnostic methods in radionuclide imaging. The results can be applied for the following objectives: (1) Radionuclide therapy will be applied in clinical practice to treat the cancer patients or other diseases in multi-center trial (2) The newly developed monoclonal antibodies and biomolecules can be used in biology, chemistry or other basic life science research (3) The new methods for the analysis of therapeutic effects, such as dosimetry, and quantitative analysis methods of radioactivity, can be applied in basic research, such as radiation oncology and radiation biology (4) The result of the project will be expected to develop the new radioimmunoassay for drug monitoring following the clinical experiments (5) Boron porphyrin has been successfully labeled with iodine. This enables the pharmacodynamic study of the boron compound in human body (6) A method to evaluate the biological effect of neutrons on tumor cells has been developed (7) The establishment of macro- and microscopic dose assessment using alpha-track autoradiography (8) Clinical application of PDT in bladder cancers, oropharyngeal cancer and skin cancer (9) Radionuclide imaging of estrogen receptor in breast cancer, lipid metabolism, gene therapy, cancers, brain function and heart disease.

  19. Tritiated thymidine autoradiographic study on the influence of sensory and sympathetic innervation on periodontal wound healing in the rat. (United States)

    Wucherpfennig, A L; Chiego, D J; Avery, J K


    Understanding of wound healing mechanisms is important in designing preventive and therapeutic approaches to inflammatory periodontal diseases, which are a major cause of dental morbidity. In this study, cell proliferation was assessed after an experimental gingival wound; this was preceded by either resection of 3 mm of the inferior alveolar nerve, total extirpation of the superior cervical ganglion, trauma to those structures or sham operations. At different times, animals were pulsed with 0.5 microCi/g body weight of tritiated thymidine; histological sections were processed for quantitative autoradiography of different compartments of the periodontium. Wounding led to a significant increase in cell proliferation in the epithelial layer, the fibroblast compartment and the periodontal ligament, but not in the alveolar crest compartment. Sympathetic denervation significantly enhanced this response in the epithelial layer, the fibroblast compartment and the alveolar crest, whereas sensory denervation only modified the response in the fibroblast layer. Thus it appears that sympathetic innervation plays an important role in the regulation of cell proliferation in the periodontium and that pharmacological modulation of sympathetic activity should be further studied as a therapeutic approach in periodontal disease.

  20. Neuroregulation of protein synthesis in odontoblasts of the first molar of the rat after wounding. (United States)

    Chiego, D J; Avery, J K; Klein, R M


    Odontoblasts respond to occlusal trauma by increased elaboration of a matrix which is subsequently calcified to form reparative dentin. The purpose of the present study was to analyze quantitatively and compare the ability of odontoblasts to synthesize collagen after wounding in rats with an intact innervation (baseline) and in rats with sensory (inferior alveolar nerve, IAN) and/or sympathetic (superior cervical ganglion, SCG) surgical denervation. Surgery was performed 7 days prior to wounding. All rats had 1 mm of enamel and dentin removed from the occlusal surface of the first mandibular molar (resected side) with the contralateral tooth serving as a control. Rats were killed 1 h after injection with 3H-proline on days 0, 5, 10 or 15 after wounding, and mandibles were removed and processed for autoradiography. Grain counts were performed over odontoblasts throughout the pulp horns for each time period and for control and experimental molars in intact (baseline) and denervated groups. When compared to the control baseline, the experimental baseline data showed increased 3H-proline uptake throughout the study with a peak at 5 days. When compared to the baseline data, IAN and SCG results demonstrated a delay or attenuation of the protein synthetic response. The results indicate that the sensory and sympathetic neural components may regulate odontoblastic response to wounding.

  1. Leptin Increases Striatal Dopamine D2 Receptor Binding in Leptin-Deficient Obese (ob/ob) Mice

    Energy Technology Data Exchange (ETDEWEB)

    Pfaffly, J.; Michaelides, M.; Wang, G-J.; Pessin, J.E.; Volkow, N.D.; Thanos, P.K.


    Peripheral and central leptin administration have been shown to mediate central dopamine (DA) signaling. Leptin-receptor deficient rodents show decreased DA D2 receptor (D2R) binding in striatum and unique DA profiles compared to controls. Leptin-deficient mice show increased DA activity in reward-related brain regions. The objective of this study was to examine whether basal D2R-binding differences contribute to the phenotypic behaviors of leptin-deficient ob/ob mice, and whether D2R binding is altered in response to peripheral leptin treatment in these mice. Leptin decreased body weight, food intake, and plasma insulin concentration in ob/ob mice but not in wild-type mice. Basal striatal D2R binding (measured with autoradiography [{sup 3}H] spiperone) did not differ between ob/ob and wild-type mice but the response to leptin did. In wild-type mice, leptin decreased striatal D2R binding, whereas, in ob/ob mice, leptin increased D2R binding. Our findings provide further evidence that leptin modulates D2R expression in striatum and that these effects are genotype/phenotype dependent.

  2. Upregulation of Cannabinoid Type 1 Receptors in Dopamine D2 Receptor Knockout Mice Is Reversed by Chronic Forced Ethanol Consumption

    Energy Technology Data Exchange (ETDEWEB)

    Thanos, P.K.; Wang, G.; Thanos, P.K.; Gopez, V.; Delis, F.; Michaelides, M.; Grand, D.K.; Wang, G.-J.; Kunos, G.; Volkow, N.D.


    The anatomical proximity of the cannabinoid type 1 (CNR1/CB1R) and the dopamine D2 receptors (DRD2), their ability to form CB1R-DRD2 heteromers, their opposing roles in locomotion, and their involvement in ethanol's reinforcing and addictive properties prompted us to study the levels and distribution of CB1R after chronic ethanol intake, in the presence and absence of DRD2. We monitored the drinking patterns and locomotor activity of Drd2+/+ and Drd2-/- mice consuming either water or a 20% (v/v) ethanol solution (forced ethanol intake) for 6 months and used the selective CB1 receptor antagonist [{sup 3}H]SR141716A to quantify CB1R levels in different brain regions with in vitro receptor autoradiography. We found that the lack of DRD2 leads to a marked upregulation (approximately 2-fold increase) of CB1R in the cerebral cortex, the caudate-putamen, and the nucleus accumbens, which was reversed by chronic ethanol intake. The results suggest that DRD2-mediated dopaminergic neurotransmission and chronic ethanol intake exert an inhibitory effect on cannabinoid receptor expression in cortical and striatal regions implicated in the reinforcing and addictive properties of ethanol.

  3. Autoradiographic study of serotonin transporter during memory formation. (United States)

    Tellez, Ruth; Rocha, Luisa; Castillo, Carlos; Meneses, Alfredo


    Serotonin transporter (SERT) has been associated with drugs of abuse like d-methamphetamine (METH). METH is well known to produce effects on the monoamine systems but it is unclear how METH affects SERT and memory. Here the effects of METH and the serotonin reuptake inhibitor fluoxetine (FLX) on autoshaping and novel object recognition (NOR) were investigated. Notably, both memory tasks recruit different behavioral, neural and cognitive demand. In autoshaping task a dose-response curve for METH was determined. METH (1.0mg/kg) impaired short-term memory (STM; lasting less of 90min) in NOR and impaired both STM and long-term memory (LTM; lasting 24 and 48h) in autoshaping, indicating that METH had long-lasting effects in the latter task. A comparative autoradiography study of the relationship between the binding pattern of SERT in autoshaping new untrained vs. trained treated (METH, FLX, or both) animals was made. Considering that hemispheric dominance is important for LTM, hence right vs. left hemisphere of the brain was compared. Results showed that trained animals decreased cortical SERT binding relative to untrained ones. In untrained and trained treated animals with the amnesic dose (1.0mg/kg) of METH SERT binding in several areas including hippocampus and cortex decreased, more remarkably in the trained animals. In contrast, FLX improved memory, increased SERT binding, prevented the METH amnesic effect and re-established the SERT binding. In general, memory and amnesia seemed to make SERT more vulnerable to drugs effects.

  4. Expression of the 5-HT receptors in rat brain during memory consolidation. (United States)

    Meneses, A; Manuel-Apolinar, L; Rocha, L; Castillo, E; Castillo, C


    Serotonin (5-hydroxytryptamine, 5-HT) system displays more than 14 receptors subtypes on brain areas involved in learning and memory processes, and pharmacological manipulation of specific receptors selectively affects memory formation. In order to begin the search of 5-HT receptors expression during memory formation, in this work, we aimed to determine, by autoradiography (using 3H 5-HT as ligand, 2 nM, specific activity 123 Ci/mmol), 5-HT receptors (5-HTR) expression in passive (untrained) and autoshaping trained (3 sessions) adult (3 months) and old (9 months) male rats. Thus, trained adult rats had better retention than old animals. Raphe nuclei of adult and old trained rats expressed less receptors on medial and dorsal, respectively. Hippocampal CA1 area and dentate gyrus of adult trained rats expressed less 5-HTR, while dentate gyrus of old increased them. Basomedial amygdaloid nucleus in old trained rats expressed more 5-HTR; while in the basolateral amygdaloid nucleus they were augmented in both groups. Training decreased or did not change 5-HTR in caudate-putamen of adult or old animals. The above profile of 5-HTR expression is consistent with previous reports, and suggests that memory formation and aging modulates 5-HTR expression in brain areas relevant to memory systems.

  5. Tachykinin receptors in the equine pelvic flexure. (United States)

    Sonea, I M; Wilson, D V; Bowker, R M; Robinson, N E


    Tachykinins, of which substance P (SP) is the prototype, are neuropeptides which are widely distributed in the nervous systems. In the equine gut, SP is present in enteric nerves and is a powerful constrictor of enteric muscle; in other species, SP is also known to have potent vasodilatory and pro-inflammatory effects. The specific effects of SP are determined by the subtype of receptor present in the target tissue. There are 3 known subtypes of tachykinin receptors, distinguished by their relative affinities for SP and other tachykinins. The distribution of SP binding sites in the equine pelvic flexure was determined using 125I-Bolton Hunter SP (I-BHSP) autoradiography. Most I-BHSP binding sites were determined to be saturable and specific, therefore presumably representing tachykinin receptors. The greatest degree of I-BHSP binding occurred over very small vessels, and over the muscularis mucosae; I-BHSP binding was also intense over the circular muscle of the muscularis externa and mucosa, and present, although less intense, over the longitudinal muscle of the muscularis externa. Competition of I-BHSP with specific receptor agonists for binding sites in the equine pelvic flexure were used to determine the subtypes of tachykinin receptors present. The neurokinin-1 receptor subtype predominated in the equine pelvic flexure, followed by the neurokinin-3 receptor subtype.

  6. Preparation of highly specific radioactivity [{sup 18}F]flumazenil and its evaluation in cynomolgus monkey by positron emission tomography

    Energy Technology Data Exchange (ETDEWEB)

    Ryzhikov, Nikolaj N. [Institute of Human Brain, Russian Academy of Science, 9, Pavlov str, 197376, Saint Petersburg (Russian Federation); Seneca, Nicholas [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden); National Institute of Mental Health, National Institutes of Health, Molecular Imaging Branch, Bethesda, MD 20892 (United States); Krasikova, Raisa N. [Institute of Human Brain, Russian Academy of Science, 9, Pavlov str, 197376, Saint Petersburg (Russian Federation); Gomzina, Natalia A. [Institute of Human Brain, Russian Academy of Science, 9, Pavlov str, 197376, Saint Petersburg (Russian Federation); Shchukin, Evgeny [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden); Fedorova, Olga S. [Institute of Human Brain, Russian Academy of Science, 9, Pavlov str, 197376, Saint Petersburg (Russian Federation); Vassiliev, Dmitrij A. [Institute of Human Brain, Russian Academy of Science, 9, Pavlov str, 197376, Saint Petersburg (Russian Federation); Gulyas, Balazs [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden); Hall, Hakan [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden); Savic, Ivanka [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden); Halldin, Christer [Department of Clinical Neuroscience, Karolinska Institutet, Psychiatry Section, Karolinska Hospital, S-17176 Stockholm (Sweden)]. E-mail:


    A straightforward method for the preparation of no-carrier-added (n.c.a.) [{sup 18}F]flumazenil via standard nucleophilic radiofluorination of the corresponding nitro-analog Ro 15-2344 has been developed. The labeling was performed by employing the K{sup 18}F/kryptofix complex in DMF at 160 deg. C for 30 min and equimolar ratio [K/K2.2.2]{sup +18}F{sup -}/precursor. Under these conditions, an {sup 18}F incorporation rate into flumazenil was in the range of 55-60%. The final product was isolated by HPLC purification within a total synthesis time of 75 min and a radiochemical yield of about 30% (EOB). Human post-mortem whole-hemisphere autoradiography of brain sections demonstrated selective uptake of the radioligand in the areas of high density of the central benzodiazepine receptors (BZR). PET studies in a cynomolgus monkey and metabolite studies by HPLC demonstrated similar results by [{sup 18}F]flumazenil as for [{sup 11}C]flumazenil. In blocking experiments, almost all radioactivity was inhibited by the addition of unlabeled flumazenil. [{sup 18}F]Flumazenil is a suitable radioligand for PET assessment of the BZR.

  7. Autoradiographic study of dopamine transporter in rat Model of Parkinson' s disease with 125I-β-CIT

    Institute of Scientific and Technical Information of China (English)

    Liu Zhenguo; Chen Shengdi; Shum Wenshan


    Objective To evaluate the value of iaaging for dopamine transpter(DAT) wi th 125I- β-CIT. Methods The partial and complete lesioned rat models of hemiparkinsonism were rendered with 6- hydroxy-dopamine (6-OHDA). Each rat was injected intravenously with 1251-β-CIT containing 40 μ Ci. Coronal t issue sections were imaged by autoradiography. The levets of dopamine (DA)and its metabolites were measured by high performance 1iquid choromatography and electro-chemical detection (HPLC-ECD). The t yros i nc hydroxylase(Tll)-positive cells and fibres in substantia nigra and striatum of the rats were observed by immunohistochemieal staining. Results The radioactivities in the lesioned striatum of both partial and complete lesioned hemiparkinsonian rats were 2.67±0.25 and O. 98±0.29 respectively , and were singificantly decreased by.18% and 72% respectively, as compared with those of unlesioned side. The levels of DA in the lesioned striatum of partial and complete lesioned models were decreased by 39% and 98% respectively. The loss of TH-positive eells and fibres in the substantia nigra and striatum was found in the lesioned striatum of both partial and complete-lesioned models. Conclusion The imaging study of DAT may be helpful for the early diagnosis of Parkinson's disease and for the monitor of the progression of this discaose;.

  8. Assessment of glutamine synthetase activity by [13N]ammonia uptake in living rat brain. (United States)

    Momosaki, Sotaro; Ito, Miwa; Tonomura, Misato; Abe, Kohji


    Glutamine synthetase (GS) plays an important role in glutamate neurotransmission or neurological disorder in the brain. [(13) N]Ammonia blood flow tracer has been reported to be metabolically trapped in the brain via the glutamate-glutamine pathway. The present study investigated the effect of an inhibitor of GS on [(13) N]ammonia uptake in order to clarify the feasibility of measuring GS activity in the living brain. l-Methionine sulfoximine (MSO), a selective GS inhibitor was microinjected into the ipsilateral striatum in rats. [(13) N]Ammonia uptake was quantified by autoradiography method as well as small animal positron emission tomography (PET) scans. The GS activity of the brain homogenate was assayed from the γ-glutamyl transferase reaction. Autoradiograms showed a decrease of [(13) N]ammonia radioactivity on the MSO-injected side compared with the saline-injected side of the striatum. This reduction could be detected with a small animal PET scanner. MSO had no effect on cerebral blood flow measured by uptake of [(15) O]H2 O. The reduction of [(13) N]ammonia uptake was closely related to the results of GS activity assay. These results indicated that [(13) N]ammonia may enable measurement of GS activity in the living brain.

  9. Absence of point mutation in the 12th codon of transformed c-Ha-rasl genes of human cancer of the breast, stomach, melanoma, and neuroblastoma

    Energy Technology Data Exchange (ETDEWEB)

    Knyazev, P.G.; Schafer, R.; Willecke, K.V.; Seitz, I.F.


    In the authors' previous investigations, they established that the tumorous cell lines SK-BR-3 (breast cancer), LAN-1 (neuroblastoma), and a heterotransplant of malignant melanoma Jal contain transforming genes of Ha-ras type. Now, the authors report their results using restriction endonucleases of MspI and HpaII restriction to study nucleotide sequences 5'-CCGGC-3' and 3'GGCCG-5', which contain the 12th codon of GGC for the amino acid glycine in the normal allele of c-Ha-rasl in the three tumors listed above, in addition to human adenocarcinoma of the stomach (CaVSt) and normal cells corresponding to them. For hybridization of MspI/HpaII, fragments of chromosomal DNA isolated from cell lines SK-BR-3, and LAN-1, Ja-1 heterotransplant, and stomach adenocarcinoma CaVSt, the XmaI section of EJ oncogene, c-Ha-rasl (plasmid pEJ 6.6), labeled with /sup 32/P was used in down-translation reaction. Hybridization was performed in 3 x SSC buffer containing 5x Deinhardt's reagent and 10% dextran sulfate at 68/sup 0/C for 16-18 h. Washing of filters was conducted under rigid conditions. For autoradiography, Kodak XR-5 x-ray film in cartridges with reinforcing shields was used at -70/sup 0/C, exposure time of four to six days.

  10. Cloned polynucleotide and synthetic oligonucleotide probes used in colony hybridization are equally efficient in the identification of enterotoxigenic Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Sommerfelt, H.; Kalland, K.H.; Raj, P.; Moseley, S.L.; Bhan, M.K.; Bjorvatn, B.


    Restriction endonuclease-generated polynucleotide and synthetically produced oligonucleotide gene probes used in colony hybridization assays proved to be efficient for the detection and differentiation of enterotoxigenic Escherichia coli. To compare their relative efficiencies, these two sets of probes were radiolabeled with /sup 32/P and were applied to 74 strains of E. coli with known enterotoxin profiles and to 156 previously unexamined E. coli isolates. The enterotoxigenic bacteria Vibrio cholerae O1, Vibrio cholerae non-O1 (NAG), Yersinia enterocolitica, and E. coli harboring the plasmid vectors of the polynucleotide gene probes were examined for further evaluation of probe specificity. The two classes of probes showed a perfect concordance in their specific detection and differentiation of enterotoxigenic E. coli. In the analysis of six strains, the signal strength on autoradiography after hybridization with oligonucleotides was weaker than that obtained after hybridization with polynucleotide probes. The probes did not hybridize with DNA from V. cholerae O1, V. cholerae non-O1 (NAG), or Y. enterocolitica. The strains of E. coli harboring the plasmid vectors of the polynucleotide gene probes were, likewise, negative in the hybridization assays.

  11. Uptake of 7,12-dimethylbenz(a)anthracene and benzo(a)pyrene in melanin-containing tissues

    Energy Technology Data Exchange (ETDEWEB)

    Roberto, A.; Larsson, B.S. [Uppsala Univ., Dept. of Pharmaceutical Biosciences, Div. of Toxicology, Uppsala (Sweden); Tjaelve, H. [The Swedish Univ. of Agricultural Sciences, Dept. of Pharmacology and Toxicology, Uppsala (Sweden)


    It is widely accepted that UV exposure is the main etiological factor for malignant melanoma. Epidemiologic studies, however, have indicated that also chemical carcinogens may be a risk factor for the disease. Polycyclic aromatic hydrocarbons such as 7,12-dimethylbenz(a)anthracene and benzo(a)pyrene represent an important class of carcinogenic chemicals. It is known that 7,12-dimethylbenz(a)anthracene can induce melanotic tumours in various animal species, and human melanocytes in culture have been found to be capable of metabolizing benzo(a)pyrene to its proximate carcinogen benzo(a)pyrene-7,8-diol. In the present study the disposition of {sup 14}C- and {sup 3}H-7,12-dimethylbenz(a)anthracene and {sup 14}C-benzo(a)pyrene was studied in pigmented and albino mice and Syrian golden hamsters by whole-body autoradiography. The results showed pronounced retention of label in the melanin-containing structures of the eyes and the hair follicles in the pigmented animals. The labelling of the corresponding structures in the albino animals was low. Additional experiments showed that 7,12-dimethylbenz(a)anthracene and benzo(a)pyrene as well as some of their metabolites are bound to melanin in vitro. The specific localization of the polycyclic aromatic hydrocarbons in pigmented tissues due to melanin affinity, combined with bioactivating capacity of melanocytes, suggest that these substances may play a role in the induction of malignant melanoma. (au).

  12. Insulin phosphorylates calmodulin in preparations of solubilized rat hepatocyte insulin receptors

    Energy Technology Data Exchange (ETDEWEB)

    Sacks, D.B.; McDonald, J.M.


    It has previously been shown that insulin stimulates the phosphorylation of calmodulin in adipocyte insulin receptor preparations. Here they demonstrate that insulin also stimulates the phosphorylation of calmodulin in wheat germ lectin-enriched insulin receptor preparations obtained from rat hepatocytes. Standard phosphorylation assays were performed at 30C in the presence of 50mM Tris-HCl (pH 7.5), 0.1% (v/v) Triton X-100, 1mM EGTA, 50 M (el-TSP)ATP, 5mM MgCl2, 0.25 M polylysine, 1.2 M calmodulin and various CaS and insulin concentrations. The phosphorylation of calmodulin was determined by SDS-PAGE and autoradiography. Phosphorylation of calmodulin had an absolute requirement for insulin receptors, insulin and certain basic proteins. Phosphorylation was maximal above 13 nM insulin and at submicromolar CaS concentrations, whereas supramicromolar CaS concentrations were inhibitory. As was observed in the adipocyte insulin receptor system, calmodulin phosphorylation was dependent upon the presence of co-factors, such as polylysine, histone H/sub f/2b and protamine sulfate. The role played by these co-factors has not yet been established. These data suggest that both CaS and calmodulin participate in post receptor insulin events in hepatocytes.

  13. Induction of a heat shock response (HSP 72) in rat embryos exposed to selected chemical teratogens. (United States)

    Mirkes, P E; Doggett, B; Cornel, L


    A monoclonal antibody to the 72 kD heat shock protein (HSP 72), Western blot analysis and 2-D gel electrophoresis/autoradiography were used to determine whether selected chemical teratogens induced the synthesis and accumulation of HSP 72 in postimplantation rat embryos exposed in vitro. The chemical teratogens studied include N-Acetoxy-2-acetylaminofluorene (N-Ac-AAF), cadmium chloride (CAD), cyclophosphamide (CP), sodium arsenite (AS), and sodium salicylate (SAL). Exposures to test chemicals were selected that produced obvious embryotoxicity characterized by abnormal development and growth retardation. Of the five chemical teratogens studied, AS and SAL induced the synthesis and accumulation of HSP 72 in day 10 rat embryos. The kinetics of HSP 72 accumulation, however, differed between AS- and SAL-treated embryos. Maximal levels of HSP 72 were observed 24 hours after AS exposure and 10 hours after SAL exposure. N-Ac-AAF, CD, and CP induced obvious embryotoxicity; however, none of these chemical teratogens induced HSP 72 at any of the timepoints assayed. Although only a small sample of chemical teratogens was studied, our results suggest that the heat shock response, characterized by the synthesis and accumulation of HSP 72, is not a general biomarker for chemical teratogens.

  14. Localization of Proliferating Cells in the Inter-Vertebral Region of the Developing and Adult Vertebrae of Lizards in Relation to Growth and Regeneration. (United States)

    Alibardi, Lorenzo


    New cartilaginous tissues in lizards is formed during the regeneration of the tail or after vertebral damage. In order to understand the origin of new cartilaginous cells in the embryo and after injury of adult vertebrae we have studied the distribution of proliferating cartilaginous cells in the vertebral column of embryos and adults of the lizard Anolis lineatopus using autoradiography for H3-thymidine and light and ultrastructural immunocytochemistry for 5BrdU. Proliferating sclerotomal cells initially surround the notochord in a segmental pattern and give rise to the chondrocytes of the vertebral centrum that replace the original chordal cells. Qualitative observations show that proliferating sclerotomal cells dilute the labeling up to 13 days post-injection but a few maintain the labeling as long labeling retention cells and remain in the inter-centra and perichondrium after birth. These cells supply new chondroblasts for post-natal growth of vertebrae but can also proliferate in case of vertebral damage or tail amputation in lizards, a process that sustains tail regeneration. The lack of somitic organization in the regenerating tail impedes the re-formation of a segmental vertebral column that is instead replaced by a continuous cartilaginous tube. It is hypothesized that long labeling retaining cells might represent stem/primordial cells, and that their permanence in the inter-vertebral cartilages and the nearby perichondrium in adult lizards pre-adapt these reptiles to elicit a broad cartilage regeneration in case of injury of the vertebrae.

  15. Original and regenerating lizard tail cartilage contain putative resident stem/progenitor cells. (United States)

    Alibardi, Lorenzo


    Regeneration of cartilaginous tissues is limited in mammals but it occurs with variable extension in lizards (reptiles), including in their vertebrae. The ability of lizard vertebrae to regenerate cartilaginous tissue that is later replaced with bone has been analyzed using tritiated thymidine autoradiography and 5BrdU immunocytochemistry after single pulse or prolonged-pulse and chase experiments. The massive cartilage regeneration that can restore broad vertebral regions and gives rise to a long cartilaginous tube in the regenerating tail, depends from the permanence of some chondrogenic cells within adult vertebrae. Few cells that retain tritiated thymidine or 5-bromodeoxy-uridine for over 35 days are mainly localized in the inter-vertebral cartilage and in sparse chondrogenic regions of the neural arch of the vertebrae, suggesting that they are putative resident stem/progenitor cells. The study supports previous hypothesis indicating that the massive regeneration of the cartilaginous tissue in damaged vertebrae and in the regenerating tail of lizards derive from resident stem cells mainly present in the cartilaginous areas of the vertebrae including in the perichondrium that are retained in adult lizards as growing centers for most of their lifetime.

  16. An effective fixative for glucocorticoid receptors in fetal tissues

    Energy Technology Data Exchange (ETDEWEB)

    Koga, T.; Kurisu, K.


    As a preliminary study in an autoradiographic study of glucocorticoid (GC) receptor localization in orofacial tissues of mouse fetuses, a search was made to determine the most effective fixative for preservation of the GC-receptor complex. Twelve-day-old mouse fetuses were administered tritiated triamcinolone acetonide (/sup 3/H-TAC) intraamniotically and subsequently processed by one of the following three procedures: freeze-drying, prefixation with Karnovsky's fixative, or the catechin fixative (Karnovsky's fixative containing 1% D-catechin) and postfixation with osmium tetroxide. Light microscopic autoradiography and liquid scintillation counting of the specimens revealed that the catechin fixative gave the best results for fixation of the steroid-receptor complex and preservation of tissue structure. Light and electron microscopic autoradiographic studies of the time course of the localization of /sup 3/H-TAC in palatal shelves supported the catechin fixative as being the most effective in preservation of GC-receptor or ligand complexes.

  17. Utility of spatially-resolved atmospheric pressure surface sampling and ionization techniques as alternatives to mass spectrometric imaging (MSI) in drug metabolism

    Energy Technology Data Exchange (ETDEWEB)

    Blatherwick, Eleanor Q. [University of Warwick, UK; Van Berkel, Gary J [ORNL; Pickup, Kathryn [AstraZeneca R& D Sweden; Johansson, Maria K. [AstraZeneca R& D Sweden; Beaudoin, Marie-Eve [AstraZeneca, USA; Cole, Roderic [ORNL; Day, Jennifer M. [AstraZeneca R& D, UK; Iverson, Suzanne [AstraZeneca R& D Sweden; Wilson, Ian D. [AstraZeneca R& D, UK; Scrivens, James H. [University of Warwick, UK; Weston, Daniel J. [AstraZeneca R& D, UK


    1. Tissue distribution studies of drug molecules play an essential role in the pharmaceutical industry and are commonly undertaken using quantitative whole body autoradiography (QWBA) methods. 2. The growing need for complementary methods to address some scientific gaps around radiography methods has led to increased use of mass spectrometric imaging (MSI) technology over the last 5 to 10 years. More recently, the development of novel mass spectrometric techniques for ambient surface sampling has redefined what can be regarded as fit-for-purpose for MSI in a drug metabolism and disposition arena. 3. Together with a review of these novel alternatives, this paper details the use of two liquid microjunction (LMJ)- based mass spectrometric surface sampling technologies. These approaches are used to provide qualitative determination of parent drug in rat liver tissue slices using liquid extraction surface analysis (LESA) and to assess the performance of a LMJ surface sampling probe (LMJ-SSP) interface for quantitative assessment of parent drug in brain, liver and muscle tissue slices. 4. An assessment of the utility of these spatially-resolved sampling methods is given, showing interdependence between mass spectrometric and QWBA methods, in particular there emerges a reason to question typical MSI workflows for drug metabolism; suggesting the expedient use of profile or region analysis may be more appropriate, rather than generating time-intensive molecular images of the entire tissue section.

  18. Implication of dorsostriatal D3 receptors in motivational processes: a potential target for neuropsychiatric symptoms in Parkinson’s disease (United States)

    Favier, Mathieu; Carcenac, Carole; Drui, Guillaume; Vachez, Yvan; Boulet, Sabrina; Savasta, Marc; Carnicella, Sebastien


    Beyond classical motor symptoms, motivational and affective deficits are frequently observed in Parkinson’s disease (PD), dramatically impairing the quality of life of patients. Using bilateral 6-hydroxydopamine (6-OHDA) lesions of the substantia nigra pars compacta (SNc) in rats, we have been able to reproduce these neuropsychiatric/non-motor impairments. The present study describes how bilateral 6-OHDA SNc lesions affect the function of the main striatal dopaminergic (DA) receptor subtypes. Autoradiography was used to measure the levels of striatal DA receptors, and operant sucrose self-administration and neuropharmacological approaches were combined to investigate the causal implication of specific DA receptors subtypes in the motivational deficits induced by a dorsostriatal DA denervation. We found that D3 receptors (D3R) exclusively are down-regulated within the dorsal striatum of lesioned rats. We next showed that infusion of a D3R antagonist (SB-277011A) in non-lesioned animals specifically disrupts preparatory, but not consummatory behaviors. Our findings reveal an unexpected involvement of dorsostriatal D3R in motivational processes. They strongly suggest an implication of dorsostriatal D3R in the neuropsychiatric symptoms observed in PD, highlighting this receptor as a potential target for pharmacological treatment. PMID:28134302

  19. The galactose-recognizing system of rat peritoneal macrophages; identification and characterization of the receptor molecule. (United States)

    Kelm, S; Schauer, R


    Resident rat peritoneal macrophages express a galactose-recognizing system, which mediates binding and uptake of cells and glycoproteins exposing terminal galactose residues. Here we describe the identification, isolation, and characterization of the corresponding receptor molecule. Using photoaffinity labelling of adherent peritoneal macrophages with the 4-azido-6-125I-salicylic acid derivative of anti-freeze glycoprotein 8 followed by SDS-PAGE and autoradiography, we identified the receptor of these cells as a protein with an apparent molecular mass of 42 kDa. Furthermore, cell surface receptors were radioiodinated by an affinity-supported labelling technique using the conjugate of asialoorosomucoid and lactoperoxidase, followed by extraction and isolation by affinity chromatography. Finally, the native receptor was isolated and analysed. To estimate its binding activity in solutions, a suitable binding assay was developed, using the precipitation of receptor-ligand complex with polyethylene glycol to separate bound from unbound 125I-asialoorosomucoid, which was used as ligand. It is shown that the isolated receptor binds to galactose-exposing particles and distinguishes between sialidase-treated and -untreated erythrocytes, similar to peritoneal macrophages. The binding characteristics of the membrane-bound and the solubilized receptor are described in the following paper of Lee et al.

  20. Recognition of Mother's voice evokes metabolic activation in the medial prefrontal cortex and lateral thalamus of Octodon degus pups. (United States)

    Braun, K; Poeggel, G


    In a variety of animal species, including primates, vocal communication is an essential part to establish and maintain social interactions, including the emotional bond between the newborn, its parents and siblings. The aim of this study in pups of the trumpet-tailed rat, Octodon degus, was to identify cortical and subcortical brain regions, which are involved in the perception of vocalizations uttered by the mother. In this species, which is characterized by an elaborated vocal repertoire, the (14C)-2-fluoro-deoxyglucose autoradiography was applied to measure region-specific metabolic activation in response to the presentation of a learned emotionally relevant acoustic stimulus, the maternal calls. Already at the age of eight days the precentral medial cortex, anterior cingulate cortex and the lateral thalamus could be identified by their enhanced metabolic activation in response to the presentation of the emotionally relevant maternal nursing calls, whereas other brain areas, such as the hippocampus and amygdala did not show stimulus-induced activation. Since in humans changes of activity patterns in relation to the emotional content of spoken language have been observed in similar brain regions, e.g. in the anterior cingulate cortex, Octodon degus may provide a suitable animal model to study the cellular and synaptic mechanisms underlying perception, production and processing of conspecific vocalizations.

  1. Differential association with cellular substructures of pseudorabies virus DNA during early and late phases of replication

    Energy Technology Data Exchange (ETDEWEB)

    Ben-Porat, T.; Veach, R.A.; Blankenship, M.L.; Kaplan, A.S.


    Pseudorabies virus DNA synthesis can be divided into two phases, early and late, which can be distinguished from each other on the basis of the structures of the replicating DNA. The two types of replicating virus DNA can also be distinguished from each other on the basis of the cellular substructures with which each is associated. Analysis by electron microscopic autoradiography showed that during the first round of replication, nascent virus DNA was found in the vicinity of the nuclear membrane; during later rounds of replication the nascent virus DNA was located centrally within the nucleus. The degree of association of virus DNA synthesized at early and late phases with the nuclear matrix fractions also differed; a larger proportion of late than of early nascent virus DNA was associated with this fraction. While nascent cellular DNA only was associated in significant amounts with the nuclear matrix fraction, a large part (up to 40%) of all the virus DNA remained associated with this fraction. However, no retention of specific virus proteins in this fraction was observed. Except for two virus proteins, which were preferentially extracted from the nuclear matrix, approximately 20% of all virus proteins remained in the nuclear matrix fraction. The large proportion of virus DNA associated with the nuclear fraction indicated that virus DNA may be intimately associated with some proteins.

  2. 99mTc(CO)3-Labeled Benzothiazole Derivatives Preferentially Bind Cerebrovascular Amyloid: Potential Use as Imaging Agents for Cerebral Amyloid Angiopathy. (United States)

    Jia, Jianhua; Cui, Mengchao; Dai, Jiapei; Liu, Boli


    Cerebral amyloid angiopathy (CAA) is a disorder affecting the elderly that is characterized by amyloid-β (Aβ) deposition in blood vessel walls of the brain. A series of 99mTc(CO)3-labeled benzothiazole derivatives as potential SPECT imaging probes for cerebrovascular Aβ deposition is reported. Rhenium surrogate displayed high affinities to Aβ aggregates with Ki values ranging from 106 to 42 nM, and they strongly stained Aβ deposits in transgenic mice (Tg) and Alzheimer's disease (AD) patients. In vitro autoradiography on brain sections of Tg and AD patients confirmed that [99mTc]24 possessed sufficient affinity for Aβ plaques, and [99mTc]24 could only label Aβ deposition in blood vessels but not Aβ plaques in the parenchyma of the brain of AD patients. Moreover, [99mTc]24 possessed favorable initial uptake (1.21% ID/g) and fast blood washout (blood2 min/blood60 min=23) in normal mice. These preliminary results suggest that [99mTc]24 may be used as an Aβ imaging probe for the detection of CAA.

  3. Replicon properties of chromosomal DNA fibers and the duration of DNA synthesis of sunflower root-tip meristem cells at different temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Van' t Hof, J.; Bjerknes, C.A.; Clinton, J.H.


    Chromosomal DNA fiber autoradiography was used to examine the replicon properties of root-tip meristem cells of Helianthus annuus intact seedlings grown at temperatures from 10 to 38/sup 0/C and those of root-tip cells grown in vitro at 23/sup 0/. The average replicon size was approximately 22 and it did not change with temperature nor when the roots were grown in culture. The average fork rate was 6 at 10/sup 0/ and it rose gradually to 12 at 38/sup 0/. The responses of replication fork movement and of the duration of S to temperature were of three types: those in which change in fork rate was primarily (more than 90%) responsible for change in the duration of S, those in which the fork rate remained constant while S increased nearly twofold, and those in which the duration of S increased even though the replication forks were moving faster. The first type of response listed was observed at temperatures from 20 to 35/sup 0/, the second type listed was observed at 10 to 15/sup 0/, and the third, was produced at 38/sup 0/.

  4. Profiling of Epstein-Barr virus latent RNA expression in clinical specimens by gene-specific multiprimed cDNA synthesis and PCR. (United States)

    Stevens, Servi J C; Brink, Antoinette A T P; Middeldorp, Jaap M


    We describe a two-step RT-PCR method for simultaneous detection of EBNA-1 (QK and Y3K splice variants), EBNA-2, LMP-1, LMP-2a and -2b, ZEBRA, and BARTs RNA encoded by Epstein-Barr virus. As a control for RNA integrity, the low-copy-number transcript derived from U1A snRNP, a cellular housekeeping gene, is coamplified. Copy DNA (cDNA) for these nine targets is simultaneously synthesized in a gene-specific, multiprimed cDNA reaction, which strongly reduces the amount of required clinical specimen and allows more sensitive detection than random hexamer or oligo-dT priming. For amplification, cDNA synthesis is followed by nine separate PCRs for the mentioned targets. Primers were designed either as intron-flanking, to avoid background DNA amplification, or in different exons, allowing identification of differentially spliced RNA molecules. To increase specificity, PCR products are detected by autoradiography after hybridization with radiolabeled internal oligonucleotide probes. The method described is highly suitable for profiling EBV latent RNA expression in tissue biopsies, cultured or isolated cells, and unfractionated whole blood and for definition of EBV latency type I, II, or III gene expression in these samples.

  5. Distribution of neurotransmitter receptors and zinc in the pigeon (Columba livia) hippocampal formation: A basis for further comparison with the mammalian hippocampus. (United States)

    Herold, Christina; Bingman, Verner P; Ströckens, Felix; Letzner, Sara; Sauvage, Magdalena; Palomero-Gallagher, Nicola; Zilles, Karl; Güntürkün, Onur


    The avian hippocampal formation (HF) and mammalian hippocampus share a similar functional role in spatial cognition, but the underlying neuronal mechanisms allowing the functional similarity are incompletely understood. To understand better the organization of the avian HF and its transmitter receptors, we analyzed binding site densities for glutamatergic AMPA, NMDA, and kainate receptors; GABAA receptors; muscarinic M1 , M2 and nicotinic (nACh) acetylcholine receptors; noradrenergic α1 and α2 receptors; serotonergic 5-HT1A receptors; dopaminergic D1/5 receptors by using quantitative in vitro receptor autoradiography. Additionally, we performed a modified Timm staining procedure to label zinc. The regionally different receptor densities mapped well onto seven HF subdivisions previously described. Several differences in receptor expression highlighted distinct HF subdivisions. Notable examples include 1) high GABAA and α1 receptor expression, which rendered distinctive ventral subdivisions; 2) high α2 receptor expression, which rendered distinctive a dorsomedial subdivision; 3) distinct kainate, α2 , and muscarinic receptor densities that rendered distinctive the two dorsolateral subdivisions; and 4) a dorsomedial region characterized by high kainate receptor density. We further observed similarities in receptor binding densities between subdivisions of the avian and mammalian HF. Despite the similarities, we propose that 300 hundred million years of independent evolution has led to a mosaic of similarities and differences in the organization of the avian HF and mammalian hippocampus and that thinking about the avian HF in terms of the strict organization of the mammalian hippocampus is likely insufficient to understand the HF of birds.

  6. Localization of glucocorticoid receptor messenger ribonucleic acid in hippocampus of rat brain using in situ hybridization

    Energy Technology Data Exchange (ETDEWEB)

    Yang, G.; Matocha, M.F.; Rapoport, S.I.


    An in situ hybridization procedure was applied to quantify glucocorticoid receptor (GR) mRNAs in the hippocampus of rat brain. Hybridization was carried out using a radiolabeled antisense probe complementary to the rat liver GR gene. The specificity of the method was validated by showing: 1) a high cellular grain density in sections hybridized with an antisense but not a sense probe; 2) agreement between the experimental and theoretical temperature at which 50% of the hybrids melted, and 3) a high signal distribution of GR mRNA in the hippocampus, a region of brain known to preferentially concentrate steroid hormones. Within the hippocampus, however, subregional differences in hybridization densities were observed. Quantitative autoradiography indicated that the average neuronal silver grain number was highest in the pyramidal cell layers of CA2 and CA4 and lowest in those of CA1 and CA3. Also, there was a significant difference in the average grain number between all of the cell fields except for that between CA2 and CA4. These results show that contiguous but neuroanatomically distinct cell fields of the hippocampus express different levels of GR transcripts, and indicate that differential regulation of GR expression occurs in subpopulations of hippocampal neurons.

  7. Spreading of hemiretinal projections in the ipsilateral tectum following unilateral enucleation: a study of optic nerve regeneration in Xenopus with one compound eye. (United States)

    Straznicky, C; Tay, D


    Right compound eyes were formed in Xenopus embryos at stages 32-33 by the fusion of two nasal (NN), two ventral (VV) or two temporal (TT) halves. Shortly after metamorphosis the optic nerve from the compound eye was sectioned and the left intact eye removed. The retinotectal projections from the compound eye to the contralateral and ipsilateral tecta were studied by [3H]proline autoradiography and electrophysiological mapping between 6 weeks and 5 months after the postmetamorphic surgery. The results showed that NN and VV eyes projected to the entire extent of both tecta. In contrast, optic fibre projection from TT eyes, although more extensive than the normal temporal hemiretinal projection, failed to cover the caudomedial portion of the tecta. The visuotectal projections in all three combinations corresponded to typical reduplicated maps to be expected from such compound eyes, where each of the hemiretinae projected across the contralateral and ipsilateral tecta in an overlapping fashion. The rapid expansion of the hemiretinal projections of the compound eyes in the ipsilateral tectum following the removal of the resident optic fibre projection suggests that tectal markers may be carried and deployed by the incoming optic fibres themselves.

  8. Novel radioiodinated {gamma}-hydroxybutyric acid analogues for radiolabeling and Photolinking of high-affinity {gamma}-hydroxybutyric acid binding sites

    DEFF Research Database (Denmark)

    Wellendorph, Petrine; Høg, Signe; Sabbatini, Paola;


    ¿-Hydroxybutyric acid (GHB) is a therapeutic drug, a drug of abuse, and an endogenous substance that binds to low- and high-affinity sites in the mammalian brain. To target the specific GHB binding sites, we have developed a (125)I-labeled GHB analog and characterized its binding in rat brain...... homogenate and slices. Our data show that [(125)I]4-hydroxy-4-[4-(2-iodobenzyloxy)phenyl]butanoate ([(125)I]BnOPh-GHB) binds to one site in rat brain cortical membranes with low nanomolar affinity (K(d), 7 nM; B(max), 61 pmol/mg protein). The binding is inhibited by GHB and selected analogs......, but not by ¿-aminobutyric acid. Autoradiography using horizontal slices from rat brain demonstrates the highest density of binding in hippocampus and cortical regions and the lowest density in the cerebellum. Altogether, the findings correlate with the labeling and brain regional distribution of high-affinity GHB sites...

  9. Novel Radioiodinated γ-Hydroxybutyric Acid Analogues for Radiolabeling and Photolinking of High-Affinity γ-Hydroxybutyric Acid Binding Sites

    DEFF Research Database (Denmark)

    Wellendorph, Petrine; Høg, Signe; Sabbatini, Paola;


    γ-Hydroxybutyric acid (GHB) is a therapeutic drug, a drug of abuse, and an endogenous substance that binds to low- and high-affinity sites in the mammalian brain. To target the specific GHB binding sites, we have developed a 125I-labeled GHB analog and characterized its binding in rat brain...... homogenate and slices. Our data show that [125I]4-hydroxy-4-[4-(2-iodobenzyloxy)phenyl]butanoate ([125I]BnOPh-GHB) binds to one site in rat brain cortical membranes with low nanomolar affinity (Kd, 7 nM; Bmax, 61 pmol/mg protein). The binding is inhibited by GHB and selected analogs, but not by γ......-aminobutyric acid. Autoradiography using horizontal slices from rat brain demonstrates the highest density of binding in hippocampus and cortical regions and the lowest density in the cerebellum. Altogether, the findings correlate with the labeling and brain regional distribution of high-affinity GHB sites or [3H...

  10. Anatomical Location of LPA1 Activation and LPA Phospholipid Precursors in Rodent and Human Brain (United States)

    González de San Román, E; Manuel, I; Giralt, MT; Chun, J; Estivill-Torrús, G; Rodriguez de Fonseca, F; Santín, LJ; Ferrer, I; Rodriguez-Puertas, R


    Lysophosphatidic acid (LPA) is a signaling molecule that binds to six known G protein-coupled receptors (GPCRs): LPA1–LPA6. LPA evokes several responses in the CNS including cortical development and folding, growth of the axonal cone and its retraction process. Those cell processes involve survival, migration, adhesion proliferation, differentiation and myelination. The anatomical localization of LPA1 is incompletely understood, particularly with regard to LPA binding. Therefore, we have used functional [35S]GTPγS autoradiography to verify the anatomical distribution of LPA1 binding sites in adult rodent and human brain. The greatest activity was observed in myelinated areas of the white matter such as corpus callosum, internal capsule and cerebellum. MaLPA1-null mice (a variant of LPA1-null) lack [35S]GTPγS basal binding in white matter areas, where the LPA1 receptor is expressed at high levels, suggesting a relevant role of the activity of this receptor in the most myelinated brain areas. In addition, phospholipid precursors of LPA were localized by MALDI-IMS in both rodent and human brain slices identifying numerous species of phosphatides (PA) and phosphatidylcholines (PC). Both PA and PC species represent potential LPA precursors. The anatomical distribution of these precursors in rodent and human brain may indicate a metabolic relationship between LPA and LPA1 receptors. PMID:25857358

  11. N-terminal galanin-(1-16) fragment is an agonist at the hippocampal galanin receptor

    Energy Technology Data Exchange (ETDEWEB)

    Fisone, G.; Berthold, M.; Bedecs, K.; Unden, A.; Bartfai, T.; Bertorelli, R.; Consolo, S.; Crawley, J.; Martin, B.; Nilsson, S.; (Univ. of Stockholm (Sweden))


    The galanin N-terminal fragment (galanin-(1-16)) has been prepared by solid-phase synthesis and by enzymic cleavage of galanin by endoproteinase Asp-N. This peptide fragment displaced {sup 125}I-labeled galanin in receptor autoradiography experiments on rat forebrain and spinal cord and in equilibrium binding experiments from high-affinity binding sites in the ventral hippocampus with an IC50 of approximately 3 nM. In tissue slices of the same brain area, galanin-(1-16), similarly to galanin, inhibited the muscarinic agonist-stimulated breakdown of inositol phospholipids. Upon intracerebroventricular administration, galanin-(1-16) (10 micrograms/15 microliters) also inhibited the scopolamine (0.3 mg/kg, s.c.)-evoked release of acetylcholine, as studied in vivo by microdialysis. Substitution of (L-Trp2) for (D-Trp2) resulted in a 500-fold loss in affinity as compared with galanin-(1-16). It is concluded that, in the ventral hippocampus, the N-terminal galanin fragment (galanin-(1-16)) is recognized by the galanin receptors controlling acetylcholine release and muscarinic agonist-stimulated inositol phospholipid breakdown as a high-affinity agonist and that amino acid residue (Trp2) plays an important role in the receptor-ligand interactions.

  12. Ca sup 2+ binding capacity of cytoplasmic proteins from rod photoreceptors is mainly due to arrestin

    Energy Technology Data Exchange (ETDEWEB)

    Huppertz, B.; Weyand, I.; Bauer, P.J. (Institut fuer Biologische Informationsverarbeitung, Forschungszentrum Juelich GmbH (Germany, F.R.))


    Arrestin (also called S-antigen or 48-kDa protein) binds to photoexcited and phosphorylated rhodopsin and, thereby, blocks competitively the activation of transducin. Using Ca{sup 2+} titration in the presence of the indicator arsenazo III and {sup 45}Ca{sup 2+} autoradiography, we show that arrestin is a Ca2(+)-binding protein. The Ca{sup 2+} binding capacity of arresting-containing protein extracts from bovine rod outer segments is about twice as high as that of arrestin-depleted extracts. The difference in the Ca{sup 2+} binding of arrestin-containing and arrestin-depleted protein extracts was attributed to arrestin. Both, these difference-measurements of protein extracts and the measurements of purified arrestin yield dissociation constants for the Ca{sup 2+} binding of arrestin between 2 and 4 microM. The titration curves are consistent with a molar ratio of one Ca{sup 2+} binding site per arrestin. No Ca{sup 2+} binding in the micromolar range was found in extracts containing mainly transducin and cGMP-phosphodiesterase. Since arrestin is one of the most abundant proteins in rod photoreceptors occurring presumably up to millimolar concentrations in rod outer segments, we suggest that aside from its function to prevent the activation of transducin, arrestin acts probably as an intracellular Ca{sup 2+} buffer.

  13. Quantitative distribution of monoamine oxidase A in brainstem monoamine nuclei is normal in major depression. (United States)

    Ordway, G A; Farley, J T; Dilley, G E; Overholser, J C; Meltzer, H Y; Balraj, E K; Stockmeier, C A; Klimek, V


    An abnormal expression of noradrenergic proteins (e.g., tyrosine hydroxylase, norepinephrine transporters) in the locus coeruleus has recently been demonstrated in subjects with major depression and/or victims of suicide. Monoamine oxidase A (MAO-A) is a key enzyme in the catabolism of biogenic amines and is expressed in brain noradrenergic neurons. In this study, the binding of [3H]Ro41-1049 to MAO-A was measured by quantitative autoradiography at multiple levels along the rostral-caudal axis of the noradrenergic locus coeruleus from subjects with major depression and age- and postmortem interval-matched control subjects who were psychiatrically normal. [3H]Ro41-1049 binding to MAO-A was unevenly distributed along the axis of the locus coeruleus, paralleling an uneven number of neuromelanin-containing (noradrenergic) neurons throughout the nucleus. Accordingly, there was a significant correlation between the number of neuromelanin-containing neurons per section and the specific binding of [3H]Ro41-1049 at any particular level of the locus coeruleus in control subjects (r(2)=0.25; pdepression (r(2)=0.14; pdepression to psychiatrically normal control subjects. These findings demonstrate that the pathophysiology of major depression is not likely to involve abnormalities in MAO-A.

  14. SIMS Analyses of Aerodynamic Fallout from a Uranium-Fueled Test

    Energy Technology Data Exchange (ETDEWEB)

    Lewis, L. A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Univ. of California, Berkeley, CA (United States); Knight, K. B. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Matzel, J. E. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Prussin, S. G. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Ryerson, F. J. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Kinman, W. S. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Zimmer, M. M. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Hutcheon, I. D. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)


    Five silicate fallout glass spherules produced in a uranium-fueled, near-surface nuclear test were characterized by secondary ion mass spectrometry, electron probe microanalysis, autoradiography, scanning electron microscopy, and energy-dispersive x ray spectroscopy. Several samples display distinctive compositional heterogeneity suggestive of incomplete mixing, and exhibit heterogeneity in U isotopes with 0.02 < 235U/ 238U < 11.8 among all five samples and 0.02 < 235U/ 238U < 7.81 within a single sample. In two samples, the 235U/ 238U ratio is correlated with major element composition, consistent with the agglomeration of chemically and isotopically distinct molten precursors. Two samples are quasi-homogeneous with respect to composition and uranium isotopic composition, suggesting extensive mixing possibly due longer residence time in the fireball. Correlated variations between 234U, 235U, 236U and 238U abundances point to mixing of end-members corresponding to uranium derived from the device and natural U ( 238U/ 235U = 0.00725) found in soil.

  15. Marked accumulation of valproic acid in embryonic neuroepithelium of the mouse during early organogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Dencker, L.; Nau, H.; D' Argy, R. (Univ. of Uppsala (Sweden))


    Valproic acid, an antiepileptic drug, causes neural tube defects in mice and man. 14C-labeled valproic acid (sodium-salt) was administered to pregnant mice on days 8 and 9 of gestation (period of high sensitivity in regard to formation of neural tube defects in this species). Two dose levels of valproic acid (1 and 400 mg/kg) were used; in each case the total radioactivity administered was the same: 400 microCi/kg or 14.7 MBq/kg. Autoradiography combined with computerized densitometry revealed that in low-dose animals most of the radioactivity was confined to maternal liver and kidney, while at high doses more activity was observed in soft tissues and fluids, including amniotic fluid. In the embryo, the neuroepithelium showed the highest concentration, irrespective of dose and survival interval (30 min, 3 h, and 6 h). Upon administration of the high dose, up to five times more radioactivity (approximately 2,000 times more valproic acid) was recovered in embryonic tissues than after the low dose. It is concluded that high doses of VPA saturate the capacities of metabolism, excretion, and protein binding in the maternal organism, resulting in a higher proportion of the dose reaching the embryo, allowing more of the drug to be accumulated by the target organ, the neuroepithelium.

  16. Crossing axons in the third nerve nucleus. (United States)

    Bienfang, D C


    The research presented in this paper studied the pathway taken by the crossed fibers of the third nerve nucleus in an animal whose nucleus has been well mapped and found to correlate well with higher mammals and man. Autoradiography using tritiated amino acid labeled the cell bodies an axons of the left side of the oculomotor nucleus of the cat. Axons so labeled could be seen emerging from the ventral portion of the left nucleus through the median longitudinal fasciculus (mlf) to join the left oculomotor nerve. Labeled axons were also seen to emerge from the medial border of the caudal left nucleus, cross the midline, and pass through the right nucleus and the right mlf to join the right oculomotor nerve. These latter axons must be the crossed axons of the superior rectus and levator palpebrae subnuclei. Since the path of these crossed axons is through the caudal portion of the nucleus of the opposite side, the destruction of one lateral half of the oculomotor nucleus would result in a bilateral palsy of the crossed subnuclei. Bilateral palsy of the superior rectus and bilateral assymetrical palsy of the levator palpebrae muscles would result.

  17. Targeting Potassium Channels for Increasing Delivery of Imaging Agents and Therapeutics to Brain Tumors

    Directory of Open Access Journals (Sweden)

    Nagendra Sanyasihally Ningaraj


    Full Text Available Every year in the US, 20,000 new primary and nearly 200,000 metastatic brain tumor cases are reported. The cerebral microvessels/ capillaries that form the blood–brain barrier (BBB not only protect the brain from toxic agents in the blood but also pose a significant hindrance to the delivery of small and large therapeutic molecules. Different strategies have been employed to circumvent the physiological barrier posed by blood-brain tumor barrier (BTB. Studies in our laboratory have identified significant differences in the expression levels of certain genes and proteins between normal and brain tumor capillary endothelial cells. In this study, we validated the non-invasive and clinically relevant Dynamic Contrast Enhancing-Magnetic Resonance Imaging (DCE-MRI method with invasive, clinically irrelevant but highly accurate Quantitative Autoradiography (QAR method using rat glioma model. We also showed that DCE-MRI metric of tissue vessel perfusion-permeability is sensitive to changes in blood vessel permeability following administration of calcium-activated potassium (BKCa channel activator NS-1619. Our results show that human gliomas and brain tumor endothelial cells that overexpress BKCa channels can be targeted for increased BTB permeability for MRI enhancing agents to brain tumors. We conclude that monitoring the outcome of increased MRI enhancing agents’ delivery to microsatellites and leading tumor edges in glioma patients would lead to beneficial clinical outcome.

  18. Autoradiographic Mapping of 5-HT1B/1D Binding Sites in the Rhesus Monkey Brain Using [carbonyl-11C]zolmitriptan

    Directory of Open Access Journals (Sweden)

    Örjan Lindhe


    Full Text Available Zolmitriptan is a serotonin 5-HT1B/1D receptor agonist that is an effective and well-tolerated drug for migraine treatment. In a human positron emission tomography study, [11C]zolmitriptan crossed the blood-brain barrier but no clear pattern of regional uptake was discernable. The objective of this study was to map the binding of [11C]zolmitriptan in Rhesus monkey brain using whole hemisphere in vitro autoradiography with [11C]zolmitriptan as a radioligand. In saturation studies, [11C]zolmitriptan showed specific (90% binding to a population of high-affinity binding sites (Kd 0.95–5.06 nM. There was regional distribution of binding sites with the highest density in the ventral pallidum, followed by the external globus pallidus, substantia nigra, visual cortex, and nucleus accumbens. In competitive binding studies with 5-HT1 receptor antagonists, [11C]zolmitriptan binding was blocked by selective 5-HT1B and 5-HT1D ligands in all target areas. There was no appreciable change in binding with the addition of a 5-HT1A receptor antagonist.

  19. Bovine gallbladder muscularis: Source of a myogenic receptor for cholecystokinin

    Energy Technology Data Exchange (ETDEWEB)

    Schjoldager, B.; Shaw, M.J.; Powers, S.P.; Schmalz, P.E.; Szurszewski, J.; Miller, L.J. (Mayo Clinic and Foundation, Rochester, MN (USA))


    Despite being a classic target for the gastrointestinal peptide hormone, cholecystokinin (CCK), the gallbladder CCK receptor is not well characterized. Pharmacological studies of small species suggest that CCK action can be mediated by direct myogenic or by both myogenic and neurogenic receptors. To prepare for the biochemical characterization of a gallbladder CCK receptor and to define the subtype of the receptor being studied. The authors have performed autoradiographic localization and pharmacological characterization of CCK receptors on bovine gallbladder. Autoradiography demonstrated high-affinity specific CCK-binding sites only on the muscularis. CCK-8 stimulated tonic contraction of longitudinal strips of gallbladder muscularis in a concentration-dependent manner. Antagonism at the cholinergic receptor with 1{mu}M atropine or axonal transmission with 1{mu}M tetrodotoxin did not modify CCK-induced contraction, supporting a direct myogenic effect of this hormone. Optimal electrical field stimulation to elicit a neuronal response resulted in muscle strip relaxation, which was abolished with adrenergic blockade. Although acetylcholine administration stimulated contraction, electrical field stimulation did not, even in the presence of phentolamine, propranolol, and/or CCK. Thus, in bovine gallbladder muscularis, there is evidence for a functional CCK receptor only on smooth muscle cells. Demonstration of a single, high-affinity specific CCK-binding site on an enriched plasma membrane preparation of bovine gallbladder muscularis is consistent with this representing a myogenic CCK receptor.

  20. Toxin a from Clostridium difficile binds to rabbit erythrocyte glycolipids with therminal Gal. cap alpha. 1-3Gal. beta. 1-4GlcNaC sequences

    Energy Technology Data Exchange (ETDEWEB)

    Clark, G.F.; Krivan, H.; Wilkins, T.; Smith, D.F.


    Toxin A is one of two clostridial toxins implicated as the causative agent of pseudomembranous colitis in patients undergoing postoperative antibiotic therapy. Evidence that the carbohydrate binding determinant for this toxin is a glycoconjugate(s) with non-reducing Gal..cap alpha..1-3Gal..beta..1-4GlcNAc has recently been reported. Specific agglutination of rabbit erythrocytes by Toxin A is inhibited by bovine thyroglobulin and prevented by pretreatment of cells with ..cap alpha..-galactosidase. Total lipid extracts from rabbit erythrocytes were subjected to thin layer chromatography and the chromatogram overlaid with purified /sup 125/I-labeled Toxin A. Two major and several minor toxin-binding glycolipids were detected following autoradiography. The major toxin-binding glycolipids were identified as pentasaccharide- and decasaccharide-ceramides expressing terminal Gal..cap alpha..1-3Gal..beta..1-4GlcNAc sequences. Treatment of the toxin-binding glycolipids with ..cap alpha..-galactosidase abolished binding. Forsmann glycolipid, globoside, Gal..cap alpha..1-4 Gal..beta..1-4Glc-cer, and Gal..cap alpha..1-3Gal..beta..1-4Glc-cer did not bind the toxin. These observations are consistent with the proposed carbohydrate specificity of the toxin for the non-reducing terminal sequence, Gal..cap alpha..1-3Gal..beta..1-4GlcNAc.

  1. Bilateral plasticity of Vibrissae SII representation induced by classical conditioning in mice. (United States)

    Debowska, Weronika; Liguz-Lecznar, Monika; Kossut, Malgorzata


    The somatosensory cortex in mice contains primary (SI) and secondary (SII) areas, differing in somatotopic precision, topographic organization, and function. The role of SII in somatosensory processing is still poorly understood. SII is activated bilaterally during attentional tasks and is considered to play a role in tactile memory and sensorimotor integration. We measured the plasticity of SII activation after associative learning based on classical conditioning, in which unilateral stimulation of one row of vibrissae was paired with a tail shock. The training consisted of three daily 10 min sessions, during which 40 pairings were delivered. Cortical activation driven by stimulation of vibrissae was mapped with 2-[(14)C]deoxyglucose (2DG) autoradiography 1 d after the end of conditioning. We reported previously that the conditioning procedure resulted in unilateral enlargement of 2DG-labeled cortical representation of the "trained" row of vibrissae in SI. Here, we measured the width and intensity of the labeled region in SII. We found that both measured parameters in SII increased bilaterally. The increase was observed in cortical layers II/III and IV. Apparently, plasticity in SII is not a simple reflection of changes in SI. It may be attributable to bilateral integrative role of SII, its lesser topographical specificity, and strong involvement in attentional processing.

  2. Advancements of labelled radio-pharmaceutics imaging with the PIM-MPGD

    Energy Technology Data Exchange (ETDEWEB)

    Donnard, J; Berny, R; Carduner, H; Leray, P; Morteau, E; Servagent, N; Thers, D [Subatech, in2p3/Ecole des Mines de Nantes/Universite de Nantes, La Chantrerie 4 rue Alfred Kastler BP 20722 44307 NANTES CEDEX 3 (France); Arlicot, N, E-mail: jerome.donnard@subatech.in2p3.f [INSERM U930 CNRS ERL 3106/Universite Francois Rabelais de Tours, CHRU Bretonneau 2 boulevard Tonnelle, 37044 TOURS CEDEX (France)


    The Beta autoradiography is widely used in pharmacology or in biological fields to study the response of an organism to a certain kind of molecule. The image of the distribution is processed by studying the concentration of the radioactivity into different organs. We report on the development of an integrated apparatus based on a PIM device (Parallel Ionization Multiplier) able to process the image of 10 microscope slides at the same time over an area of 18*18 cm{sup 2}. Thanks to a vacuum pump and a regulation gas circuit, 5 minutes is sufficient to begin an acquisition. All the electronics and the gas distribution are included in the structure leading to a transportable device. Special software has been developed to process data in real time with image visualization. Biological samples can be labelled with beta emitters of low energy like {sup 3}H/{sup 14}C or Auger electrons of {sup 125}I/{sup 99m}Tc. The measured spatial resolution is 30 mum in {sup 3}H and the trigger and the charge rate are constant over more than 6 days of acquisition showing good stability of the device. Moreover, collaboration with doctors and biologists of INSERM (National Institute for Medical Research in France) has started in order to demonstrate that MPGD's can be easily proposed outside a physics laboratory.

  3. Eimeria tenella: parasite-specific incorporation of /sup 3/H-uracil as a quantitative measure of intracellular development

    Energy Technology Data Exchange (ETDEWEB)

    Schmatz, D.M.; Crane, M.S.; Murray, P.K.


    An assay has been developed using parasite-specific incorporation of /sup 3/H-uracil to assess the intracellular growth of Eimeria tenella in vitro. As shown by both scintillation counts and autoradiography, /sup 3/H-uracil was incorporated specifically into intracellular parasites from the onset of infection and continued throughout development of the first generation schizonts. Mature schizonts and first generation merozoites did not continue to incorporate additional /sup 3/H-uracil, indicating that RNA synthesis had halted in these stages. Based on these findings, a semi-automated microscale uracil incorporation assay was developed to determine parasite viability. This method should be useful for biochemical studies with intracellular parasites and for screening compounds for anticoccidial activity. The ease, rapidity, and quantitative nature of this assay contrasts favorably with standard morphometric approaches of determining parasite development. In addition, parallel studies using host cell incorporation of /sup 3/H-uridine have been introduced as a method of determining whether antiparasitic activity is direct or indirect in relation to effects on the host cell.

  4. Studies on the in vitro cultivation of coccidia

    Energy Technology Data Exchange (ETDEWEB)

    Schmatz, D.M.


    New approaches to the in vitro cultivation of coccidian parasites are described here, specifically for avian coccidia of the genus Eimeria. Firstly, an improved method of purifying the infectious stage of these parasites, known as sporozoites, over a DEAE-52 cellulose anion exchange column to eliminate toxic debris generated during excystation is described. The cultured cells used to support the intracellular development of these parasites, Madin-Darby Bovine Kidney Cells (MDBK), were cloned and it was demonstrated that some clones were more susceptible than others to infection with sporozoites. The use of sub-lethal doses of gamma radiation to pre-treat host cell monolayers prior to infecting has been found to prevent host cell overgrowth and subsequent peeling of the monolayers while not interfering with parasite development. Utilizing in vitro culture techniques developed here in conjunction with radiolabeling studies, an assay has been development using the parasite-specific incorporation of /sup 3/H-uracil to assess the intracellular development of E. tenella and E. acervulina in vitro. As shown by both scintillation counts and autoradiography, /sup 3/H-uracil was incorporated specifically into the intracellular parasites from the onset of infection and continued throughout the development of the first generation schizonts. Based on these findings, a semi-automated microscale incorporation assay was developed to determine parasite viability. The assay system is used in this study to investigate the effects of known anticoccidials, sporozoite antiserum, and varying the composition of the cell culture medium on parasite development.

  5. Ultrastructural study of the chromatoid body in planarian regenerative cells

    Energy Technology Data Exchange (ETDEWEB)

    Hori, I. (Kanazawa Medical Univ., Ishikawa (Japan))


    The present paper deals with the ultrastructural changes of chromatoid bodies in planarian regenerative cells under normal and experimental conditions. A close relationship was usually observed between chromatoid bodies and pore regions of the nuclear envelope in these cells. The chromatoid bodies continued to decrease in size during cytodifferentiation of regenerative cells, though they did not disappear entirely throughout the regeneration processes. Cytochemistry and (/sup 3/H)uridine autoradiography have shown that the chromatoid body contains RNA. The typical morphological effect of actinomycin D became apparent in three organelles, i.e., nucleolus, polysome and chromatoid body. Ultrastructural changes in nucleoli were observed to occur after actinomycin treatment (20 The exposure to a higher dose of actinomycin (50 caused a decay of chromatoid bodies while nuclear envelopes retained numerous pores. Both the nucleoli and the chromatoid bodies disappeared in the sequential stages. Within the cytoplasm of such cells disintegration of a polysomal pattern was correlated with the disappearance of chromatoid bodies. The significance of the planarian chromatoid body is discussed in relation to differentiation of the regenerative cells.

  6. BETAview: a digital {beta}-imaging system for dynamic studies of biological phenomena

    Energy Technology Data Exchange (ETDEWEB)

    Bertolucci, E.; Conti, M.; Mettivier, G.; Montesi, M.C. E-mail:; Russo, P


    We present a digital autoradiography (DAR) system, named BETAview, based on semiconductor pixel detectors and a single particle counting chip, for quantitative analysis of {beta}-emitting radioactive tracers in biological samples. The system is able to perform a real time monitoring of time-dependent biological phenomena. BETAview could be equipped either with GaAs or with Si semiconductor pixellated detectors. In this paper, we describe the results obtained with an assembly based on a Si detector, 300 {mu}m thick, segmented into 64x64 170 {mu}m size square pixels. The detector is bump-bonded to the low threshold, single particle counting chip named Medipix1, developed by a CERN-based European collaboration. The sensitive area is about 1 cm{sup 2}. Studies of background noise and detection efficiency have been performed. Moreover, time-resolved cellular uptake studies with radiolabelled molecules have been monitored. Specifically, we have followed in vivo and in real time, the [{sup 14}C]L-leucine amino acid uptake by eggs of Octopus vulgaris confirming the preliminary results of a previous paper. This opens the field of biomolecular kynetic studies with this new class of semiconductor DAR systems, whose evolution (using the Medipix2 chip, 256x256 pixels, 55 {mu}m pixel size) is soon to come.

  7. Luminescence imaging using radionuclides: a potential application in molecular imaging

    Energy Technology Data Exchange (ETDEWEB)

    Park, Jeong Chan [Department of Molecular Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Il An, Gwang [Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul 139-706 (Korea, Republic of); Park, Se-Il [Department of Molecular Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Oh, Jungmin [Korea Basic Science Institute Chuncheon Center, Gangwon-do 200-701 (Korea, Republic of); Kim, Hong Joo [Department of Physics and Energy Science, Kyungpook National University, Daegu 702-710 (Korea, Republic of); Su Ha, Yeong; Wang, Eun Kyung [Department of Molecular Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Min Kim, Kyeong; Kim, Jung Young [Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul 139-706 (Korea, Republic of); Lee, Jaetae [Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Welch, Michael J. [Department of Radiology, Washington University School of Medicine, St. Louis, MO 63110 (United States); Yoo, Jeongsoo, E-mail: [Department of Molecular Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of)


    Introduction: Nuclear and optical imaging are complementary in many aspects and there would be many advantages when optical imaging probes are prepared using radionuclides rather than classic fluorophores, and when nuclear and optical dual images are obtained using single imaging probe. Methods: The luminescence intensities of various radionuclides having different decay modes have been assayed using luminescence imaging and in vitro luminometer. Radioiodinated Herceptin was injected into a tumor-bearing mouse, and luminescence and microPET images were obtained. The plant dipped in [{sup 32}P]phosphate solution was scanned in luminescence mode. Radio-TLC plate was also imaged in the same imaging mode. Results: Radionuclides emitting high energy {beta}{sup +}/{beta}{sup -} particles showed higher luminescence signals. NIH3T6.7 tumors were detected in both optical and nuclear imaging. The uptake of [{sup 32}P]phosphate in plant was easily followed by luminescence imaging. Radio-TLC plate was visualized and radiochemical purity was quantified using luminescence imaging. Conclusion: Many radionuclides with high energetic {beta}{sup +} or {beta}{sup -} particles during decay were found to be imaged in luminescence mode due mainly to Cerenkov radiation. 'Cerenkov imaging' provides a new optical imaging platform and an invaluable bridge between optical and nuclear imaging. New optical imaging probes could be easily prepared using well-established radioiodination methods. Cerenkov imaging will have more applications in the research field of plant science and autoradiography.

  8. Hyperglycolysis is exacerbated after traumatic brain injury with fentanyl vs. isoflurane anesthesia in rats. (United States)

    Statler, Kimberly D; Janesko, Keri L; Melick, John A; Clark, Robert S B; Jenkins, Larry W; Kochanek, Patrick M


    Despite common use of narcotics in the clinical management of severe traumatic brain injury (TBI), in experimental models rats treated with fentanyl have exhibited worse functional outcome and more CA1 hippocampal death than rats treated with standard isoflurane anesthesia. We hypothesized that greater post-traumatic excitotoxicity, reflected by cerebral glucose utilization (CMRglu), may account for detrimental effects of fentanyl vs. isoflurane. Rats were anesthetized with either isoflurane (1% by inhalation) or fentanyl (10 mcg/kg iv bolus then 50 mcg/kg/h infusion). 14C-deoxyglucose autoradiography was performed 45 min after controlled cortical impact (CCI) to left parietal cortex (n=4 per anesthetic group) or in uninjured rats after 45 min of anesthesia (n=3 per anesthetic group). Uninjured rats treated with fentanyl vs. isoflurane showed 35-45% higher CMRglu in all brain structures (panesthesia. This post-traumatic hyperglycolysis suggests greater excitotoxicity and concurs with reports of worse functional outcome and more CA1 hippocampal death after TBI with fentanyl vs. isoflurane anesthesia.

  9. Epithelial binding of 1,1,2,2-tetrachloroethane in the respiratory and upper alimentary tract

    Energy Technology Data Exchange (ETDEWEB)

    Eriksson, C.; Brittebo, E.B. (Swedish Univ. of Agricultural Sciences, Uppsala (Sweden). Dept. of Pharmacology and Toxicology)


    The bioactivation and binding of {sup 14}C-labelled 1,1,2,2-tetrachloroethane (TCE) in the tissues of C57B1 mice were studied. As shown by autoradiography with heated and organic solvent-extracted tissue sections of i.v. injected mice, a high and selective localization of bound metabolites occurred in the nasal olfactory mucosa, preferentially in the Bowman's glands. High levels of bound metabolites were also present in epithelia of the trachea, bronchi and bronchioli and in the squamous epithelia of the oral cavity, tongue and esophagus. An epithelial binding was observed in tissue slices incubated with {sup 14}C-TCE. Incubation of {sup 14}C-TCE with homogenates of the olfactory mucosa and liver showed that the olfactory mucosa had a higher ability to activate {sup 14}C-TCE into products that become irreversibly bound to protein. Addition of metyrapone, glutathione or sodium dithionite to the incubations decreased the level of irreversible binding, suggesting that the activation of TCE to reactive products is mediated via an oxidative cytochrome P-450 dependent process in the olfactory mucosa. (orig.).

  10. Boron microquantification in oral mucosa and skin following administration of a neutron capture therapy agent

    Energy Technology Data Exchange (ETDEWEB)

    Kiger, S.W. III; Micca, P.L.; Morris, G.M.; Coderre, J.A


    Clinical trials of boron neutron capture therapy (BNCT) for intracranial tumours using boronphenylalanine-fructose undertaken at Harvard-MIT and Brookhaven National Laboratory have observed acute normal tissue reactions in the skin and oral mucosa. Because the range of the {sup 10}B(n,a){sup 7}Li reaction products is very short, 10-14 {mu}m combined, knowledge of the 10B microdistribution in tissue is critical for understanding the microdosimetry and radiobiology of BNCT. This paper reports measurements of the microdistribution of {sup 10}B in an animal model, rat skin and tongue, using high resolution quantitative autoradiography (HRQAR), a neutron-induced track etch autoradiographic technique. The steep spatial gradient and high absolute value relative to blood of the {sup 10}B concentration observed in some strata of the rat tongue epithelium and skin are important for properly evaluating the radiobiology and the biological effectiveness factors for normal tissue reactions such as oral mucositis, which are generally assessed using the blood boron concentration rather than the tissue boron concentration. (author)

  11. DNA synthesis index: higher for human gallbladders with cholesterol gallstones than with pigment gallstones

    Energy Technology Data Exchange (ETDEWEB)

    Lamote, J.; Putz, P.; Francois, M.; Willems, G.


    (/sup 3/H)dThd uptake by the gallbladder epithelium was estimated in 33 patients with cholesterol stones, in 13 patients with pigment stones, and in 12 gallbladders without stones. Proliferative parameters were estimated by autoradiography after in vitro incubation with (/sup 3/H)-dThd. Stones were identified by quantitative infrared spectroscopy. The degree of inflammation of the gallbladder wall was estimated by a histologic scoring method. In the gallbladders containing cholesterol stones the DNA synthesis index (1.39 +/- 0.28%) was higher (P less than .01) than in the gallbladders without stones (0.19 +/- 0.04%). No significant increase in proliferative parameters was found in the gallbladders with pigment stones (0.24 +/- 0.06%). No correlation was found between total stone number, weight or volume, and the DNA synthesis index. No evidence was observed that inflammation could influence the epithelial cell proliferation. Something in the bile of patients with cholesterol stones rather than the physical presence of stones may be the cause of the variations observed.

  12. Imaging of peripheral-type benzodiazepine receptor in tumor: carbon ion irradiation reduced the uptake of a positron emission tomography ligand [11C]DAC in tumor. (United States)

    Yamasaki, Tomoteru; Koike, Sachiko; Hatori, Akiko; Yanamoto, Kazuhiko; Kawamura, Kazunori; Yui, Joji; Kumata, Katsushi; Ando, Koichi; Zhang, Ming-Rong


    We aimed to determine the effect of carbon ion irradiation on the uptake of N-benzyl-N-11C-methyl-2-(7-methyl-8-oxo-2-phenyl-7,8-dihydro-9H-purin-9-yl)acetamide ([(11)C]DAC), a positron emission tomography (PET) ligand for the peripheral-type benzodiazepine receptor (PBR), in tumor cells and tumor-bearing mice. Spontaneous murine fibrosarcoma (NFSa) cells were implanted into the right hind legs of syngeneic C3H male mice. Conditioning irradiation with 290 MeV/u carbon ions was delivered to the 7- to 8-mm tumors In vitro uptake of [(11)C]DAC was measured in single NFSa cells isolated from NFSa-bearing mice after irradiation. In vivo biodistribution of [(11)C]DAC in NFSa-bearing mice was determined by small animal PET scanning and dissection. In vitro autoradiography was performed using tumor sections prepared from mice after PET scanning. In vitro and in vivo uptake of [(11)C]DAC in single NFSa cells and NFSa-bearing mice was significantly reduced by carbon ion irradiation. The decrease in [(11)C]DAC uptake in the tumor sections was mainly due to the change in PBR expression. In conclusion, [(11)C]DAC PET responded to the change in PBR expression in tumors caused by carbon ion irradiation in this study. Thus, [(11)C]DAC is a promising predictor for evaluating the effect of carbon ion radiotherapy.

  13. Novel use of positively charged nylon transfer membranes for trapping indoleacetic acid or other small anions during efflux from plant tissues (United States)

    Evans, M. L.; Hangarter, R. P.


    Positively charged nylon blotting membranes were used as an anion binding medium to trap [14C]indoleactic acid (IAA) as it exited cells at the basal ends of Coleus blumei L. stem and Zea mays L. coleoptile segments. Autoradiography was used to visualize where the [14C] that moved out of the cut ends was localized on the nylon membrane. Diffusion of [14C]IAA from the initial point of contact with the nylon membrane was minimal. Comparison of the autoradiograms with anatomical tissue prints of the cut ends of the segments was used to determine what tissues participate in IAA movement. The results of these initial studies were consistent with other reports suggesting that [14C]IAA movement was primarily associated with vascular tissues in both C. blumei stems and corn coleoptiles, but the resolution was not sufficient to identify which vascular tissues were involved in IAA transport. With further refinements, this technique could also be used for studying the movement of other small charged molecules through plant tissues.

  14. In-situ removal and characterisation of uranium-containing particles from sediments surrounding the Fukushima Daiichi Nuclear Power Plant (United States)

    Martin, P. G.; Griffiths, I.; Jones, C. P.; Stitt, C. A.; Davies-Milner, M.; Mosselmans, J. F. W.; Yamashiki, Y.; Richards, D. A.; Scott, T. B.


    Traditional methods to locate and subsequently study radioactive fallout particles have focused heavily on autoradiography coupled with in-situ analytical techniques. Presented here is the application of a Variable Pressure Scanning Electron Microscope with both backscattered electron and energy dispersive spectroscopy detectors, along with a micromanipulator setup and electron-hardening adhesive to isolate and remove individual particles before synchrotron radiation analysis. This system allows for a greater range of new and existing analytical techniques, at increased detail and speed, to be applied to the material. Using this method, it was possible to erform detailed energy dispersive spectroscopy and synchrotron radiation characterisation of material likely ejected from the Fukushima Daiichi Nuclear Power Plant found within a sediment sample collected from the edge of the 30 km exclusion zone. Particulate material sub-micron in maximum dimension examined during this work via energy dispersive spectroscopy was observed to contain uranium at levels between 19.68 and 28.35 weight percent, with the application of synchrotron radiation spectroscopy confirming its presence as a major constituent. With great effort and cost being devoted to the remediation of significant areas of eastern Japan affected by the incident, it is crucial to gain the greatest possible understanding of the nature of this contamination in order to inform the most appropriate clean-up response.

  15. Development of a New Positron Emission Tomography Tracer for Targeting Tumor Angiogenesis: Synthesis, Small Animal Imaging, and Radiation Dosimetry

    Directory of Open Access Journals (Sweden)

    David S. Lalush


    Full Text Available Angiogenesis plays a key role in cancer progression and correlates with disease aggressiveness and poor clinical outcomes. Affinity ligands discovered by screening phage display random peptide libraries can be engineered to molecularly target tumor blood vessels for noninvasive imaging and early detection of tumor aggressiveness. In this study, we tested the ability of a phage-display-selected peptide sequence recognizing specifically bone marrow- derived pro-angiogenic tumor-homing cells, the QFP-peptide, radiolabeled with 64Cu radioisotope to selectively image tumor vasculature in vivo by positron emission tomography (PET. To prepare the targeted PET tracer we modified QFP-phage with the DOTA chelator and radiolabeled the purified QFP-phage-DOTA intermediate with 64Cu to obtain QFP-targeted radioconjugate with high radiopharmaceutical yield and specific activity. We evaluated the new PET tracer in vivo in a subcutaneous (s.c. Lewis lung carcinoma (LLC mouse model and conducted tissue distribution, small animal PET/CT imaging study, autoradiography, histology, fluorescence imaging, and dosimetry assessments. The results from this study show that, in the context of the s.c. LLC immunocompetent mouse model, the QFP-tracer can target tumor blood vessels selectively. However, further optimization of the biodistribution and dosimetry profile of the tracer is necessary to ensure efficient radiopharmaceutical applications enabled by the biological specificity of the QFP-peptide.

  16. Comparative targeting of human colon-carcinoma multicell spheroids using one- and two-step (bispecific antibody) techniques. (United States)

    Devys, A; Thedrez, P; Gautherot, E; Faivre-Chauvet, A; Saï-Maurel, C; Rouvier, E; Auget, J L; Barbet, J; Chatal, J F


    In the perspective of radioimmunotherapy (RIT) of micrometastases, we compared, in multicell spheroids (MS), the uptake and retention kinetics of 125I-F(ab)'2 F6 anti-carcinoembryonic antigen (CEA) monoclonal antibody (MAb), and the affinity enhancement system (AES) using an anti-CEA/anti-DTPA-indium bispecific antibody (BsMAb) and a 125I-labeled di-DTPA-In-tyrosine-lysine bivalent hapten. We used MS of colorectal tumor cell lines expressing CEA strongly (LS 174T), weakly (HT-29) or not at all (HRT-18). Uptake and retention kinetics of 125I-F(ab)'2 F6 and 125I-BsMAb used alone gave similar results. The highest uptake values, obtained with LS 174T MS, were slightly lower with AES than with 125I-F(ab)'2 F6. However, effective retention half-lives were longer for AES than for 125I-F(ab)'2 F6 or for 111In-labeled monovalent hapten after pre-incubation of spheroids with BsMAb. Autoradiography showed the same slow and heterogeneous distribution of 125I-F(ab)'2 F6 and 125I-BsMAb. These results indicate that the 2-step technique is more favorable for RIT: uptake values were approximately the same but uptake kinetics were more rapid, and retention half-life was longer than with the one-step technique.

  17. Sex differences in adrenocortical structure and function. Pt. 11

    Energy Technology Data Exchange (ETDEWEB)

    Malendowicz, L.K.; Jachimowicz, B.


    Adrenal glands from orchectomized and ovariectomized rats, with and without replacement therapy, and also from intact controls of both sexes, were examined by autoradiography with /sup 3/H-thymidine. The labelling index after 1 or 2 nucleoside injections was higher in the zona glomerulosa of females than in male rats, while no differences were found in the fascicular and reticular zones. Orchiectomy increased the labelling index in the fascicular and reticular zones, an effect prevented by testosterone. Ovariectomy did not change the labelling index, while estradiol lowered it in the zona glomerulosa. Duration of the S phase was longer in the zona fasciculata cells of males than in females. Both orchiectomy and testosterone shortened this phase in cells of the zona fasciculata and zona reticularis. Ovariectomy prolonged the S phase in the zona fasciculata and shortened this time in the reticular zone, an effect reversed by estradiol. In the glomerular and fascicular zones, cell cycle time was longer in males than in females. Orchiectomy shortened this time in all adrenocortical zones, an effect reversed by testosterone. Ovariectomy shortened cell cycle time in the glomerular and reticular zones and prolonged it in the zona fasciculata; these effects were reversed by estradiol. Turnover rate in adrenocortical cells was markedly higher in females than in males, a difference due to testosterone which markedly decreased turnover rate.

  18. Autoradiographic study of the effect of 1,25-dihydroxyvitamin D/sub 3/ on bone matrix synthesis in vitamin D replete rats

    Energy Technology Data Exchange (ETDEWEB)

    Hock, J.M.; Kream, B.E.; Raisz, G.


    An autoradiographic technique using pulse labels of (/sup 3/H)proline was developed to assess the early effects of 1,25-dihydroxyvitamin D/sub 3/ (1,25(OH)/sub 2/D/sub 3/) on bone matrix synthesis in vitamin D replete rats. Rats, 7 days old, were given 0.25, 2.5, or 25 ng of 1,25(OH)/sub 2/D/sub 3/ or vehicle alone subcutaneously on days 1, 3, and 5 of the experiment. Rats received a subcutaneous injection of 100 (/sup 3/H)proline on days 2 and 6 and were killed on day 7. Calvaria and tibia were processed for autoradiography, and morphometric methods were developed to measure the rate and amount of bone matrix formed during the experimental period. When compared to control values, the amount and rate of formation of new bone matrix were both significantly decreased in rats receiving 25 ng of 1,25(OH)/sub 2/D/sub 3/ and slightly, but not significantly, decreased in rats receiving 2.5 ng. We conclude that administration of pharmacologic doses of 1,25(OH)/sub 2/D/sub 3/ to vitamin D replete rat pups impairs the formation of collagenous bone matrix.

  19. The effect of vitamin A on the migration and DNA synthesis of rat bladder tumor cell line NBT II in culture. (United States)

    Tchao, R; Leighton, J


    In the presence of vitamin A, NBT II cells, derived from a carcinoma of rat bladder, grew as a monolayer with diminished piling up. Keratinization, which normally appeared within stratified cells in postconfluent cultures, was inhibited. A "wounding" technique suitable for quantitative analysis of cell migration was developed for confluent cultures grown on glass coverslips. Vitamin A treatment enhanced the migration of cells from the wound edge. In dense postconfluent monolayer cultures, vitamin A treatment maintained a higher percentage of cells in DNA synthesis than in the control cultures, as determined by 3H-TdR uptake and autoradiography. In contrast, in sparse cultures vitamin A did not stimulate DNA synthesis or increase the mitotic index. This stimulatory effect, limited to dense cultures, may be attributable to vitamin A causing viable cells to be shed into the medium, thereby maintaining the monolayer just at confluence. Thus vitamin A inhibits squamous cell differentiation, enhances migration, and maintains the culture in the proliferative phase. In a different system of high cell density, NBT II aggregates cultured in a combined matrix of chick plasma clot and collagen-coated sponge, vitamin A also enhanced the migration of cells. These results may explain, in part, the failure of vitamin A to inhibit completely the growth of some established tumors.

  20. The pharmacokinetics, distribution and degradation of human recombinant interleukin 1 beta in normal rats

    DEFF Research Database (Denmark)

    Reimers, J; Wogensen, L D; Welinder, B


    Based upon in vivo rat experiments it was recently suggested that interleukin 1 in the circulation may be implicated in the initial events of beta-cell destruction leading to insulin-dependent diabetes mellitus (IDDM) in humans. The aim of the present study was to estimate half-lives of distribut......Based upon in vivo rat experiments it was recently suggested that interleukin 1 in the circulation may be implicated in the initial events of beta-cell destruction leading to insulin-dependent diabetes mellitus (IDDM) in humans. The aim of the present study was to estimate half...... the circulation with a T1/2 alpha of 2.9 min and a T1/2 beta of 41.1 min. The central and peripheral volume of distribution was 20.7 and 19.1 ml/rat, respectively, and the metabolic clearance rate was 16.9 ml/min/kg. The kidney and liver showed the highest accumulation of tracer, and autoradiography demonstrated...

  1. Improved targeting of 5-[125I/131I]iodo-2‧-deoxyuridine to rat hepatoma by using lipiodol emulsion (United States)

    Yu, Hung-Man; Yeh, Hsin-Pei; Chang, Tien-Kui; Huang, Kuang-Liang; Chuang, Kuo-Tang; Liu, Ren-Shen; Wang, Shyh-Jen; Hwang, Jeng-Jong; Chi, Kwan-Hwa; Chen, Fu-Du; Lin, Wuu-Jyh; Chen, Chin-Hsiung; Wang, Hsin-Ell


    This study aims to assess whether emulsion of [ 125/131I]IUdR and lipiodol (IUdR/LP) can improve delivery of IUdR into hepatoma. MethodsIn vitro release profile of IUdR from IUdR/LP to serum was performed. IUdR/LP was injected into N1-S1 hepatoma-bearing SD rat via hepatic artery and IUdR/normal saline (IUdR/NS) was used for comparison. Biodistribution, autoradiography, imaging and tumor DNA incorporation assay were performed. The radioactive metabolites in plasma and urine were analyzed. Radiation doses to tumor and organs were estimated. ResultsIUdR released from lipiodol into serum was fast. There were longer retention, more DNA incorporation and higher radiation dose of IUdR in the tumor by using IUdR/LP. IUdR/LP deposited deep in the hepatomas. Only free iodide was found in the plasma and urine after injection of IUdR/LP. ConclusionsHepatic artery injection of IUdR/LP emulsion could definitely enhance the tumor cell uptake and incorporation to DNA of *IUdR, prolong the tumor retention time and increase radiation dose to tumor. IUdR/LP may be an effective therapeutic agent for the treatment of hepatic tumors.

  2. Raman microprobe investigation of molecular structure and organization in the native state of woody tissue

    Energy Technology Data Exchange (ETDEWEB)

    Atalla, R.H.


    Although the primary emphasis of our program has remained with the application of Raman spectroscopy to the study of native tissue, the scope of the work has been expanded to include a number of complementary approaches. These have included Solid State 13C NMR, autoradiography of radiolabeled woody tissue sections, and the generation of biomimetic tertiary aggregates which simulate states of aggregation characteristic of cell walls. Our Raman spectroscopic studies have resulted in progress in the areas of interpretation of the spectral features, and confirmation of the variability of the patterns of orientation of lignin reported earlier. We have assembled and made operational our new microprobe and spectrometer systems acquired under the DOE-URIP program. We have also demonstrated that, operating with gated detection and pulsed laser excitation, we can discriminate against the laser-excited fluorescence characteristic of most woody tissue. Our studies of celluloses, which combine Raman spectroscopy and 13C NMR have shown that all native celluloses are composites of two forms which have the same secondary structure but different tertiary structures.

  3. Raman microprobe investigation of molecular structure and organization in the native state of woody tissue. Progress report, April 1, 1987--July 31, 1989

    Energy Technology Data Exchange (ETDEWEB)

    Atalla, R.H.


    Although the primary emphasis of our program has remained with the application of Raman spectroscopy to the study of native tissue, the scope of the work has been expanded to include a number of complementary approaches. These have included Solid State 13C NMR, autoradiography of radiolabeled woody tissue sections, and the generation of biomimetic tertiary aggregates which simulate states of aggregation characteristic of cell walls. Our Raman spectroscopic studies have resulted in progress in the areas of interpretation of the spectral features, and confirmation of the variability of the patterns of orientation of lignin reported earlier. We have assembled and made operational our new microprobe and spectrometer systems acquired under the DOE-URIP program. We have also demonstrated that, operating with gated detection and pulsed laser excitation, we can discriminate against the laser-excited fluorescence characteristic of most woody tissue. Our studies of celluloses, which combine Raman spectroscopy and 13C NMR have shown that all native celluloses are composites of two forms which have the same secondary structure but different tertiary structures.

  4. Sexing the human fetus and identification of polyploid nuclei by DNA-DNA in situ hybridisation in interphase nuclei. (United States)

    West, J D; Gosden, C M; Gosden, J R; West, K M; Davidson, Z; Davidson, C; Nicolaides, K H


    Samples of human adult lymphocytes, fetal lymphocytes, amniotic fluid cells, and chorionic villus cells were sexed independently by cytogenetics and DNA-DNA in situ hybridisation to a tritiated Y probe. For the in situ hybridisation analysis, the presence of Y bodies (hybridisation bodies) in 100 interphase nuclei were scored after autoradiography. In all, 82/83 samples were sexed in this way (one technical failure) and 78/82 were sexed by both in situ hybridisation and cytogenetics. There was complete agreement between the two methods. There was a considerable variation (40-100%) in the percentage of interphase nuclei with a hybridisation body among the male samples, but very few nuclei from female samples showed significant hybridisation. In situ hybridisation could be used to sex the conceptus when males but not females are at risk for various X-linked genetic disorders and may also be useful for detecting 45,X/46,XY mosaicism or polyploid/diploid mosaicism. This would be particularly useful for direct preparations of chorionic villus samples, which often prove difficult to analyse cytogenetically but offer the best means of avoiding maternal contamination. Some interphase nuclei had more than one hybridisation body, and this was most commonly found among amniotic fluid cells. Comparison of sizes of nuclei with one or two hybridisation bodies strongly suggested that most of the amniotic fluid cell nuclei with two hybridisation bodies were tetraploid.

  5. Breakdown of the normal optic nerve head blood-brain barrier following acute elevation of intraocular pressure in experimental animals. (United States)

    Radius, R L; Anderson, D R


    Five hours of elevated intraocular pressure produced evidence of an altered blood-brain barrier at the optic nerve head in 27 of 29 monkey eyes. The change in vascular permeability was documented by fluorescein angiography (18 of 21 eyes), by Evans blue fluorescence microscopy (21 of 23 eyes), or by both methods. Leakage occurred from major blood vessels as well as from microvasculature of the nerve head. In 22 eyes, rapid axonal transport was studied after intravitreal injection of tritiated leucine. In 18 of these 22 eyes, autoradiography demonstrated a local interruption of axonal transport. In 15 eyes examined by all three methods, leakage from microvasculature (as opposed to leakage from the major vessels) was loosely associated with severe and widespread blockade of axonal transport at the lamina cribrosa. Although cause-and-effect relationships are not proved, ischemia may be responsible both for the focal endothelial damage with breakdown of the normal blood-brain barrier and for the local abnormalities of axonal transport.

  6. Physical Matrix Characterisation: Studies of Crystalline Rocks and Consolidated Clays by PMMA Method and Electron Microscopy as Support of Diffusion Analyses

    Energy Technology Data Exchange (ETDEWEB)

    Siitari-Kauppi, M.; Leskinen, A.; Kelokaski, A.; Togneri, L.; Alonso, U.; Missana, T.; Garcia-Gutierrez, M.; Patelli, A.


    Crystalline rock and consolidated clay are both considered adequate host rocks for a high-level radioactive waste deep geological repository (HLWR). Over the extended periods of HLWR operation, long-lived radionuclides (RN) may be released from the fuel and migrate to the geo/biosphere. To predict the fate of contaminants and to assess the safety of the host rock, it is very important to determine transport parameters, as diffusion coefficients, and to relate them to the physical properties of the barriers, as connected porosity. In heterogeneous materials, it is also a major task to describe the transport at the mineral scale evaluating diffusion coefficients and mineral-specific porosities on single minerals. The main objective of this study was to determine within granite and consolidated clay, the connective porosity and mineral-specific porosities by poly-methylmethacrylate (PMMA) autoradiography method. Scanning electron microscopy and energy-dispersive X-ray analyses (FESEM/EDS) were performed in order to study the pore apertures of porous regions in greater detail and to identify the corresponding minerals. By the novel application of the nuclear ion beam technique Rutherford Backscattering Spectrometry (RBS) apparent diffusion coefficients were determined at mineral scale. Finally, the porosity results were used to evaluate the effective diffusion coefficients and retention parameters of single minerals in different granite samples and consolidated clays. (Author) 42 refs.

  7. The studies on neurogenesis indufced by brain injury in adult ring dove

    Institute of Scientific and Technical Information of China (English)



    It was the first time demonstrated by us that the number of newborn neurons was increased after making lesion in forebrain of adult ring dove(Streptopelia risoria) by means of autoradiography and imunohistochemistry,Neurogensis in the adult avian is restricted to the telencephalon.In doves with bilateral electrolytic lesion of nucleus ectostriatum(E),the mean mumber of proliferating cells in the lateral ventricular zone(LVZ) and newborn neurons in the forebrain increased by 1.95 times and 2.38 times respectively as compared with that in intact doves.The most remarkable incresase of neurogenesis induced by nucleus ectostriatum lesions was found at the anteriorposterior level 3(L3),where the lesion site was located.These results showed that the electrolytic brain lesion al tered the distribution pattern of proliferating cells in the LVZ and resulted in increase of the number of newborn neureons in the non-VZ areas of forebrain.The changes in number and distribution pattern of proliferating cells in LVZ and newbon neurons in forebrain may be dependent on site of lesion.Studies on the relationship between proliferating cells in LVZ and newly generated neurons in non-VZ aresa may help to understand the mechanism of brain plasticity and development.

  8. Effects of insulin on hexose transport across blood-brain barrier in normoglycemia

    Energy Technology Data Exchange (ETDEWEB)

    Namba, H.; Lucignani, G.; Nehlig, A.; Patlak, C.; Pettigrew, K.; Kennedy, C.; Sokoloff, L.


    The effects of insulin on 3-O-(/sup 14/C) methylglucose transport across the blood-brain barrier (BBB) were studied in conscious rats under steady-state normoglycemic conditions. The (/sup 14/C)methylglucose was infused intravenously at a constant rate, and animals were killed at various times between 5 and 30 min after the initiation of the infusion. The time course of the arterial plasma concentration of (/sup 14/C)methylglucose was determined in timed arterial blood samples taken during the infusion. Local cerebral tissue concentrations of (/sup 14/C)methylglucose at the time of killing were determined by quantitative autoradiography of brain sections. The rate constants for inward and outward transport of (/sup 14/C)methylglucose across the BBB, K/sub 1/, and k/sub 2/, respectively, were estimated by a least-squares, best-fit of a kinetic equation to the measured time courses of plasma and tissue concentrations. The equilibrium distribution ration, K/sub 1//k/sub 2/, for (/sup 14/C)methylglucose in brain increased by approx. 10-11% in the hyperinsulinemic animals. Because 3-O-(/sup 14/C)methylglucose shares the same carrier that transports glucose and other hexoses across the BBB, these results suggest that hyperinsulinemia decreases the rate constants for transport but increases the distribution space for hexoses in brain. These effects are, however, quite small and are probably minor or negligible when compared with the major effects of insulin in other tissues.

  9. Intestinal inhibition of the Na+/H+ exchanger 3 prevents cardiorenal damage in rats and inhibits Na+ uptake in humans. (United States)

    Spencer, Andrew G; Labonte, Eric D; Rosenbaum, David P; Plato, Craig F; Carreras, Christopher W; Leadbetter, Michael R; Kozuka, Kenji; Kohler, Jill; Koo-McCoy, Samantha; He, Limin; Bell, Noah; Tabora, Jocelyn; Joly, Kristin M; Navre, Marc; Jacobs, Jeffrey W; Charmot, Dominique


    The management of sodium intake is clinically important in many disease states including heart failure, kidney disease, and hypertension. Tenapanor is an inhibitor of the sodium-proton (Na(+)/H(+)) exchanger NHE3, which plays a prominent role in sodium handling in the gastrointestinal tract and kidney. When administered orally to rats, tenapanor acted exclusively in the gastrointestinal tract to inhibit sodium uptake. We showed that the systemic availability of tenapanor was negligible through plasma pharmacokinetic studies, as well as autoradiography and mass balance studies performed with (14)C-tenapanor. In humans, tenapanor reduced urinary sodium excretion by 20 to 50 mmol/day and led to an increase of similar magnitude in stool sodium. In salt-fed nephrectomized rats exhibiting hypervolemia, cardiac hypertrophy, and arterial stiffening, tenapanor reduced extracellular fluid volume, left ventricular hypertrophy, albuminuria, and blood pressure in a dose-dependent fashion. We observed these effects whether tenapanor was administered prophylactically or after disease was established. In addition, the combination of tenapanor and the blood pressure medication enalapril improved cardiac diastolic dysfunction and arterial pulse wave velocity relative to enalapril monotherapy in this animal model. Tenapanor prevented increases in glomerular area and urinary KIM-1, a marker of renal injury. The results suggest that therapeutic alteration of sodium transport in the gastrointestinal tract instead of the kidney--the target of current drugs--could lead to improved sodium management in renal disease.

  10. Autoradiographic Mapping of 5-HT(1B/1D) Binding Sites in the Rhesus Monkey Brain Using [carbonyl-C]zolmitriptan. (United States)

    Lindhe, Orjan; Almqvist, Per; Kågedal, Matts; Gustafsson, Sven-Åke; Bergström, Mats; Nilsson, Dag; Antoni, Gunnar


    Zolmitriptan is a serotonin 5-HT(1B/1D) receptor agonist that is an effective and well-tolerated drug for migraine treatment. In a human positron emission tomography study, [(11)C]zolmitriptan crossed the blood-brain barrier but no clear pattern of regional uptake was discernable. The objective of this study was to map the binding of [(11)C]zolmitriptan in Rhesus monkey brain using whole hemisphere in vitro autoradiography with [(11)C]zolmitriptan as a radioligand. In saturation studies, [(11)C]zolmitriptan showed specific (90%) binding to a population of high-affinity binding sites (Kd 0.95-5.06 nM). There was regional distribution of binding sites with the highest density in the ventral pallidum, followed by the external globus pallidus, substantia nigra, visual cortex, and nucleus accumbens. In competitive binding studies with 5-HT(1) receptor antagonists, [(11)C]zolmitriptan binding was blocked by selective 5-HT(1B) and 5-HT(1D) ligands in all target areas. There was no appreciable change in binding with the addition of a 5-HT(1A) receptor antagonist.

  11. Species differences in 3-methylsulphonyl-DDE bioactivation by adrenocortical tissue. (United States)

    Lindström, Veronica; Brandt, Ingvar; Lindhe, Orjan


    The CYP11B1-activated adrenocortical toxicant 3-methylsulphonyl-DDE (3-MeSO2-DDE) is proposed as a lead compound for an improved chemotherapy for adrenocortical carcinoma. We compared the binding of 3-MeSO2-[14C]DDE in the adrenal cortex of four rodent species; hamster, guinea pig, mouse and rat, using a precision-cut adrenal slice culture system ex vivo. Localization and quantification of the bound radioactivity were carried out using light microscopy autoradiography and radioluminography. The results revealed major species differences since 3-MeSO2-[14C]DDE was extensively bound to the hamster adrenal tissue while the guinea pig adrenals were devoid of binding. A high binding in mouse adrenal cortex was confirmed while binding in rat adrenal cortex was very weak. The results support previous observations that metabolic activation of 3-MeSO2-DDE is highly species dependent. Since CYP11B1 could be expressed in tissues other than the adrenal cortex, final toxicological characterization should be carried out in a species that can bioactivate this compound.

  12. Improved targeting of 5-[{sup 125}I/{sup 131}I]iodo-2'-deoxyuridine to rat hepatoma by using lipiodol emulsion

    Energy Technology Data Exchange (ETDEWEB)

    Yu, H.-M. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China); Yeh, H.-P. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China); Chang, T.-K. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China); Huang, K.-L. [Institute of Nuclear Energy Research, Taoyuan, Taiwan (China); Chuang, Kuo-Tang [Institute of Nuclear Energy Research, Taoyuan, Taiwan (China); Liu, R.-S. [Department of Nuclear Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Wang, S.-J. [Department of Nuclear Medicine, Taipei Veterans General Hospital, Taipei, Taiwan (China); Hwang, J.-J. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China); Chi, K.-H. [Division of Radiation Therapy and Oncology, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan (China); Chen, F.-D. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China): Central Taiwan University of Science and Technology, Taichung, Taiwan (China); Lin, W.-J. [Institute of Nuclear Energy Research, Taoyuan, Taiwan (China); Chen, Chin-Hsiung [Institute of Nuclear Energy Research, Taoyuan, Taiwan (China); Wang, H.-E. [Institute of Radiological Sciences, National Yang-Ming University, Taipei, Taiwan (China)]. E-mail:


    This study aims to assess whether emulsion of [{sup 125/131}I]IUdR and lipiodol (IUdR/LP) can improve delivery of IUdR into hepatoma. Methods: In vitro release profile of IUdR from IUdR/LP to serum was performed. IUdR/LP was injected into N1-S1 hepatoma-bearing SD rat via hepatic artery and IUdR/normal saline (IUdR/NS) was used for comparison. Biodistribution, autoradiography, imaging and tumor DNA incorporation assay were performed. The radioactive metabolites in plasma and urine were analyzed. Radiation doses to tumor and organs were estimated. Results: IUdR released from lipiodol into serum was fast. There were longer retention, more DNA incorporation and higher radiation dose of IUdR in the tumor by using IUdR/LP. IUdR/LP deposited deep in the hepatomas. Only free iodide was found in the plasma and urine after injection of IUdR/LP. Conclusions: Hepatic artery injection of IUdR/LP emulsion could definitely enhance the tumor cell uptake and incorporation to DNA of *IUdR, prolong the tumor retention time and increase radiation dose to tumor. IUdR/LP may be an effective therapeutic agent for the treatment of hepatic tumors.

  13. 2-Phenylbenzothiazole conjugated with cyclopentadienyl tricarbonyl [CpM(CO)3] (M = Re, (99m)Tc) complexes as potential imaging probes for β-amyloid plaques. (United States)

    Jia, Jianhua; Cui, Mengchao; Dai, Jiapei; Liu, Boli


    Technetium-99m-labeled cyclopentadienyl tricarbonyl complexes conjugated with the 2-phenylbenzothiazole binding motif were synthesized. The rhenium surrogates , , and were demonstrated to have moderate to high affinities for Aβ1-42 aggregates with Ki values of 142, 76, 64 and 24 nM, respectively. During the fluorescence staining of brain sections of transgenic mice and patients with Alzheimer's disease, these rhenium complexes demonstrated perfect and intense labeling of Aβ plaques. Moreover, in in vitro autoradiography, (99m)Tc-labeled complexes clearly detected β-amyloid plaques on sections of brain tissue from transgenic mice, which confirmed the sufficient affinity of these tracers for Aβ plaques. However, these compounds did not show desirable properties in vivo, especially showing poor brain uptake (below 0.5% ID g(-1)), which will hinder the further development of these tracers as brain imaging agents. Nonetheless, it is encouraging that these (99m)Tc-labeled complexes designed by a conjugate approach displayed sufficient affinities for Aβ plaques.

  14. Visualization and Quantitative Assessment of the Brain Distribution of Insulin through Nose-to-Brain Delivery Based on the Cell-Penetrating Peptide Noncovalent Strategy. (United States)

    Kamei, Noriyasu; Shingaki, Tomotaka; Kanayama, Yousuke; Tanaka, Misa; Zochi, Riyo; Hasegawa, Koki; Watanabe, Yasuyoshi; Takeda-Morishita, Mariko


    Our recent work suggested that intranasal coadministration with the cell-penetrating peptide (CPP) penetratin increased the brain distribution of the peptide drug insulin. The present study aimed to distinctly certify the ability of penetratin to facilitate the nose-to-brain delivery of insulin by quantitatively evaluating the distribution characteristics in brain using radioactive (64)Cu-NODAGA-insulin. Autoradiography and analysis using a gamma counter of brain areas demonstrated that the accumulation of radioactivity was greatest in the olfactory bulb, the anterior part of the brain closest to the administration site, at 15 min after intranasal administration of (64)Cu-NODAGA-insulin with l- or d-penetratin. The brain accumulation of (64)Cu-NODAGA-insulin with penetratin was confirmed by ELISA using unlabeled insulin in which intact insulin was delivered to the brain after intranasal coadministration with l- or d-penetratin. By contrast, quantification of cerebrospinal fluid (CSF) samples showed increased insulin concentration in only the anterior portion of the CSF at 15 min after intranasal coadministration with l-penetratin. This study gives the first concrete proof that penetratin can accelerate the direct transport of insulin from the nasal cavity to the brain parenchyma. Further optimization of intranasal administration with CPP may increase the efficacy of delivery of biopharmaceuticals to the brain while reducing the risk of systemic drug exposure.

  15. Accumulation of radioactivity in rat brain and peripheral tissues including salivary gland after intravenous administration of {sup 14}C-D-aspartic acid

    Energy Technology Data Exchange (ETDEWEB)

    Imai, Kazuhiro; Fukushima, Takeshi; Santa, Tomofumi; Homma, Hiroshi [Tokyo Univ. (Japan). Faculty of Pharmaceutical Sciences; Sugihara, Juko; Kodama, Hirohiko; Yoshikawa, Masayoshi


    After the intravenous administration of {sup 14}C-D-aspartic acid (Asp) into Sprague-Dawley rats (male, 7-week-old), the distribution and elimination of radioactivity was investigated by the whole body autoradiography. High radioactivities were detected in pineal gland, pituitary gland and salivary gland at 30 min after administration. The other tissues detected were liver, lung, adrenal gland, pancreas and spleen where D-Asp was reported to occur naturally. After 24 hr, the radioactivities were still detected at high levels in the pineal, pituitary and salivary glands. The data suggested the natural occurrence of D-Asp in salivary gland. After careful examination utilizing fluorescent derivatization and chiral separation by high-performance liquid chromatography, the presence of D-Asp was, for the first time, demonstrated in salivary gland in situ, the concentration of which was 7.85 {+-} 1.0 nmol/g. The administration of {sup 14}C-L-Asp was also carried out. The data suggested that D-Asp in the circulating blood is one of the sources of the tissue D-Asp. (author)

  16. Connections of the corticomedial amygdala in the golden hamster. II. Efferents of the ''olfactory amygdala''

    Energy Technology Data Exchange (ETDEWEB)

    Kevetter, G.A.; Winans, S.S.


    The anterior cortical (C1) and posterolateral cortical (C2) nuclei of the amygdala are designated the ''olfactory amygdala'' because they each receive direct projections from the main olfactory bulb. The efferents of these nuclei were traced after stereotaxic placement of 1-5 muCi tritiated proline in the corticomedial amygdala of the male golden hamsters. Following survival times of 12, 24, or 48 hours, 20 micron frozen sections of the brains were processed for light microscopic autoradiography. Efferents from C2 terminate in layers II and III of the olfactory tubercle and in layer Ib of pars ventralis and pars medialis of the anterior olfactory nucleus. Fibers from this nucleus also project to layers I and II of the infralimbic cortex and to the molecular layer of the agranular insular cortex. More posteriorly, fibers from C2 terminate in layer I of the dorsolateral entorhinal cortex, and in the endopiriform nucleus. From C1, efferent fibers travel in the stria terminalis and terminate in the precommissural bed nucleus of the stria terminalis and in the mediobasal hypothalamus. Efferents from C1 also innervate the molecular layer of C2, the amygdalo-hippocampal area, and the adjacent piriform cortex. Neurons in both C1 and C2 project to the molecular layer of the medial amygdaloid nucleus and the posteromedial cortical nucleus of the amygdala, the plexiform layer of the ventral subiculum, and the molecular layer of the lateral entorhinal cortex.

  17. Distribution of /sup 58/Co from the oral cavity of mice

    Energy Technology Data Exchange (ETDEWEB)

    Stenberg, T. (Department of Prosthetics and Biophysical Laboratory, Faculty of Odontology, University of Umeaa, Umeaa, Sweden)


    Whole body retention and tissue distribution of /sup 58/Co in mice having had an oral cavity deposit of /sup 58/CoCl/sub 2/ was studied at 1, 5 and 24 h after administration by means of gamma scintillation measurements and autoradiography. The whole body /sup 58/Co retention at 5 h after administration was 93.3% of the whole body /sup 58/Co content at 1 h. Corresponding /sup 58/Co-retention at 24 h after administration was 9.2%. Tissues analyzed by gamma scintillation measurements at 1, 5 and 24 h post administration were: serum, kidney, pancreas, spleen, liver, heart, lung, tongue and palatal mucosa. At all survival times after administration palatal mucosa showed the highest /sup 58/Co median content followed by tongue. The very high /sup 58/Co median content of palatal mucosa and tongue at 1 h after administration showed a strong decrease at 5 and 24 h after administration. For the other tissues the highest /sup 58/Co median content was found in the liver and the lowest in the spleen with the highest /sup 58/Co median value for each tissue at 5 h after administration. The autoradiograms revealed a high /sup 58/Co uptake in oral, nasal, pharyngeal and esophageal mucosa, intestines and stomach. In liver, pancreas and kidney no uptake was observed at 1 h and a weak uptake was detected at 5 and 24 h.

  18. Autoradiographic localization of a gluten peptide during organ culture of human duodenal mucosa

    Energy Technology Data Exchange (ETDEWEB)

    Fluge, G.; Aksnes, L.


    An 125I-labeled subfraction of Frazer's fraction III (molecular weight, 8,000) was added to the culture medium during organ culture of duodenal biopsies from two patients with celiac disease in exacerbation. The isotope-labeled gluten peptide was localized by autoradiography after 6, 12, and 24 h of culture. At 6 h, labeling was located mainly in the basal layers of the biopsies. The tissue was well preserved. After 12 h in culture, the labeling had spread to the lamina propria and the crypts. A few grains were located over enterocytes and desquamated cells. Moderate histological signs of toxicity were observed. After 24 h, there was marked toxic deterioration, comparable to that seen after culture with alpha-gliadin. Labeling had spread throughout the entire section. There seemed to be no specificity of the binding, for the entire section was affected. Culture with the identical gluten fraction, in the radionegative state, produced histological deterioration comparable to that seen after exposure to the isotope-labeled peptide. Gluten peptides are presented to the target cells in a unique way during organ culture, different from in vivo conditions. This may influence the results when the organ culture method is used to investigate the pathogenesis of celiac disease.

  19. Effect of time period after boric acid injection on {sup 10}B absorption in different regions of adult male rat's brain

    Energy Technology Data Exchange (ETDEWEB)

    Baghban Khojasteh, Nasrin, E-mail: [Nuclear Engineering Department, Science and Research Branch, Islamic Azad University, Poonak Sq. PO Box 14515-775, Tehran (Iran, Islamic Republic of); Pazirandeh, Ali [Nuclear Engineering Department, Science and Research Branch, Islamic Azad University, Poonak Sq. PO Box 14515-775, Tehran (Iran, Islamic Republic of); Jameie, Behnam [Nuclear Engineering Department, Science and Research Branch, Islamic Azad University, Poonak Sq. PO Box 14515-775, Tehran (Iran, Islamic Republic of); Laboratory of Basic Science and Neuroscience, Basic Science Dept, Faculty of Allied Medicine, Cellular and Molecular Research Center, Tehran University of Medical Science, Pardis-e-Hemmat,Tehran (Iran, Islamic Republic of); Goodarzi, Samereh [Nuclear Engineering Department, Science and Research Branch, Islamic Azad University, Poonak Sq. PO Box 14515-775, Tehran (Iran, Islamic Republic of)


    Distribution of {sup 10}B in different regions of rat normal brain was studied. Two groups were chosen as control and trial. Trial group received 2 ml of neutral boron compound. 2, 4 and 6 h after the injection brain removed, coronal sections of forebrain, midbrain and hindbrain were sandwiched between two pieces of polycarbonate. Autoradiography plots of {sup 10}B distribution showed significant differences in three regions with the highest {sup 10}B concentration in the forebrain during 4 h after injection. - Highlights: Black-Right-Pointing-Pointer Normal tissue tolerance is very important in BNCT. Black-Right-Pointing-Pointer This study has been done to determine {sup 10}B distribution in three anatomical regions of rat normal brain. Black-Right-Pointing-Pointer These specific regions of brain have not been studied in previous BNCT projects. Black-Right-Pointing-Pointer We found significant differences in {sup 10}B distribution between these three regions. Black-Right-Pointing-Pointer In different time periods after neutral boron compound injection, there has been a significant difference in boron absorption.

  20. PET imaging of cannabinoid type 2 receptors with [(11)C]A-836339 did not evidence changes following neuroinflammation in rats. (United States)

    Pottier, Geraldine; Gómez-Vallejo, Vanessa; Padro, Daniel; Boisgard, Raphaël; Dollé, Frédéric; Llop, Jordi; Winkeler, Alexandra; Martín, Abraham


    Cannabinoid type 2 receptors (CB2R) have emerged as promising targets for the diagnosis and therapy of brain pathologies. However, no suitable radiotracers for accurate CB2R mapping have been found to date, limiting the investigation of the CB2 receptor expression using positron emission tomography (PET) imaging. In this work, we report the evaluation of the in vivo expression of CB2R with [(11)C]A-836339 PET after cerebral ischemia and in two rat models of neuroinflammation, first by intrastriatal LPS and then by AMPA injection. PET images and in vitro autoradiography showed a lack of specific [(11)C]A-836339 uptake in these animal models demonstrating the limitation of this radiotracer to image CB2 receptor under neuroinflammatory conditions. Further, using immunohistochemistry, the CB2 receptor displayed a modest expression increase after cerebral ischemia, LPS and AMPA models. Finally, [(18)F]DPA-714-PET and immunohistochemistry demonstrated decreased neuroinflammation by a selective CB2R agonist, JWH133. Taken together, these findings suggest that [(11)C]A-836339 is not a suitable radiotracer to monitor in vivo CB2R expression by using PET imaging. Future studies will have to investigate alternative radiotracers that could provide an accurate binding to CB2 receptors following brain inflammation.

  1. Region-selective effects of neuroinflammation and antioxidant treatment on peripheral benzodiazepine receptors and NMDA receptors in the rat brain

    Energy Technology Data Exchange (ETDEWEB)

    Biegon, A.; Alvarado, M.; Budinger, T.F.; Grossman, R.; Hensley, K.; West, M.S.; Kotake, Y.; Ono, M.; Floyd, R.A.


    Following induction of acute neuroinflammation by intracisternal injection of endotoxin (lipopolysaccharide) in rats, quantitative autoradiography was used to assess the regional level of microglial activation and glutamate (NMDA) receptor binding. The possible protective action of the antioxidant phenyl-tert-butyl nitrone in this model was tested by administering the drug in the drinking water for 6 days starting 24 hours after endotoxin injection. Animals were killed 7 days post-injection and consecutive cryostat brain sections labeled with [3H]PK11195 as a marker of activated microglia and [125I]iodoMK801 as a marker of the open-channel, activated state of NMDA receptors. Lipopolysaccharide increased [3H]PK11195 binding in the brain, with the largest increases (2-3 fold) in temporal and entorhinal cortex, hippocampus, and substantia innominata. A significant (>50 percent) decrease in [125I]iodoMK801 binding was found in the same brain regions. Phenyl-tert-butyl nitrone treatment resulted in a partial inhibition ({approx}25 percent decrease) of the lipopolysaccharide-induced increase in [3H]PK11195 binding but completely reversed the lipopolysaccharide-induced decrease in [125I]iodoMK80 binding in the entorhinal cortex, hippocampus, and substantia innominata. Loss of NMDA receptor function in cortical and hippocampal regions may contribute to the cognitive deficits observed in diseases with a neuroinflammatory component, such as meningitis or Alzheimer's disease.

  2. Major integral membrane protein immunogens of Treponema pallidum are proteolipids

    Energy Technology Data Exchange (ETDEWEB)

    Chamberlain, N.R.; Brandt, M.E.; Erwin, A.L.; Radolf, J.D.; Norgard, M.V. (Univ. of Texas Southwestern Medical Center, Dallas (USA))


    A number of the major pathogen-specific immunogens of Treponema pallidum were characterized recently as amphiphilic, integral membrane proteins by phase partitioning with Triton X-114. In the present study, we demonstrated that the same membrane immunogens (designated as detergent phase proteins (DPPs)) become radiolabeled upon in vitro incubation of T. pallidum with various {sup 3}H-labeled fatty acids. Radioimmunoprecipitation with a monoclonal antibody confirmed that the {sup 3}H-labeled 47-kilodalton protein corresponded to the well-characterized treponemal antigen with the identical apparent molecular mass. Failure to detect {sup 3}H-labeled DPPs following incubation with erythromycin confirmed that protein acylation required de novo protein synthesis by the bacteria. When treponemes were incubated with ({sup 3}H)myristate, ({sup 3}H)palmitate, or ({sup 3}H)oleate, radiolabeled proteins corresponding to the DPPs were detected upon autoradiography. Demonstration that a number of the abundant membrane immunogens of T. pallidum are proteolipids provides information to help clarify their membrane association(s) and may serve to explain their extraordinary immunogenicity.

  3. Synthesis and Preliminary Evaluation of a 2-Oxoquinoline Carboxylic Acid Derivative for PET Imaging the Cannabinoid Type 2 Receptor

    Directory of Open Access Journals (Sweden)

    Linjing Mu


    Full Text Available Cannabinoid receptor subtype 2 (CB2 has been shown to be up-regulated in activated microglia and therefore plays an important role in neuroinflammatory and neurodegenerative diseases such as multiple sclerosis, amyotrophic lateral sclerosis and Alzheimer’s disease. The CB2 receptor is therefore considered as a very promising target for therapeutic approaches as well as for imaging. A promising 2-oxoquinoline derivative designated KP23 was synthesized and radiolabeled and its potential as a ligand for PET imaging the CB2 receptor was evaluated. [11C]KP23 was obtained in 10%–25% radiochemical yield (decay corrected and 99% radiochemical purity. It showed high stability in phosphate buffer, rat and mouse plasma. In vitro autoradiography of rat and mouse spleen slices, as spleen expresses a high physiological expression of CB2 receptors, demonstrated that [11C]KP23 exhibits specific binding towards CB2. High spleen uptake of [11C]KP23 was observed in dynamic in vivo PET studies with Wistar rats. In conclusion, [11C]KP23 showed promising in vitro and in vivo characteristics. Further evaluation with diseased animal model which has higher CB2 expression levels in the brain is warranted.

  4. Cannabinoid receptor localization in brain

    Energy Technology Data Exchange (ETDEWEB)

    Herkenham, M.; Lynn, A.B.; Little, M.D.; Johnson, M.R.; Melvin, L.S.; de Costa, B.R.; Rice, K.C. (National Institute of Mental Health, Bethesda, MD (USA))


    (3H)CP 55,940, a radiolabeled synthetic cannabinoid, which is 10-100 times more potent in vivo than delta 9-tetrahydrocannabinol, was used to characterize and localize a specific cannabinoid receptor in brain sections. The potencies of a series of natural and synthetic cannabinoids as competitors of (3H)CP 55,940 binding correlated closely with their relative potencies in several biological assays, suggesting that the receptor characterized in our in vitro assay is the same receptor that mediates behavioral and pharmacological effects of cannabinoids, including human subjective experience. Autoradiography of cannabinoid receptors in brain sections from several mammalian species, including human, reveals a unique and conserved distribution; binding is most dense in outflow nuclei of the basal ganglia--the substantia nigra pars reticulata and globus pallidus--and in the hippocampus and cerebellum. Generally high densities in forebrain and cerebellum implicate roles for cannabinoids in cognition and movement. Sparse densities in lower brainstem areas controlling cardiovascular and respiratory functions may explain why high doses of delta 9-tetrahydrocannabinol are not lethal.

  5. Synthesis and evaluation of ethyleneoxylated and allyloxylated chalcone derivatives for imaging of amyloid β plaques by SPECT. (United States)

    Fuchigami, Takeshi; Yamashita, Yuki; Haratake, Mamoru; Ono, Masahiro; Yoshida, Sakura; Nakayama, Morio


    We report radioiodinated chalcone derivatives as new SPECT imaging probes for amyloid β (Aβ) plaques. The monoethyleneoxy derivative 2 and allyloxy derivative 8 showed a high affinity for Aβ(1-42) aggregates with Ki values of 24 and 4.5 nM, respectively. Fluorescent imaging demonstrated that 2 and 8 clearly stained thioflavin-S positive Aβ plaques in the brain sections of Tg2576 transgenic mice. In vitro autoradiography revealed that [(125)I]2 displayed no clear accumulation toward Aβ plaques in the brain sections of Tg2576 mice, whereas the accumulation pattern of [(125)I]8 matched with the presence of Aβ plaques both in the brain sections of Tg2576 mice and an AD patient. In biodistribution studies using normal mice, [(125)I]2 showed preferable in vivo pharmacokinetics (4.82%ID/g at 2 min and 0.45%ID/g at 60 min), while [(125)I]8 showed only a modest brain uptake (1.62%ID/g at 2 min) with slow clearance (0.56%ID/g at 60 min). [(125)I]8 showed prospective binding properties for Aβ plaques, although further structural modifications are needed to improve the blood brain barrier permeability and washout from brain.

  6. A Very High Spatial Resolution Detector for Small Animal PET

    Energy Technology Data Exchange (ETDEWEB)

    Kanai Shah, M.S.


    Positron Emission Tomography (PET) is an in vivo analog of autoradiography and has the potential to become a powerful new tool in imaging biological processes in small laboratory animals. PET imaging of small animals can provide unique information that can help in advancement of human disease models as well as drug development. Clinical PET scanners used for human imaging are bulky, expensive and do not have adequate spatial resolution for small animal studies. Hence, dedicated, low cost instruments are required for conducting small animal studies with higher spatial resolution than what is currently achieved with clinical as well as dedicated small animal PET scanners. The goal of the proposed project is to investigate a new all solid-state detector design for small animal PET imaging. Exceptionally high spatial resolution, good timing resolution, and excellent energy resolution are expected from the proposed detector design. The Phase I project was aimed at demonstrating the feasibility of producing high performance solid-state detectors that provide high sensitivity, spatial resolution, and timing characteristics. Energy resolution characteristics of the new detector were also investigated. The goal of the Phase II project is to advance the promising solid-state detector technology for small animal PET and determine its full potential. Detectors modules will be built and characterized and finally, a bench-top small animal PET system will be assembled and evaluated.

  7. Characteristic expression of γ-aminobutyric acid and glutamate decarboxylase in rat jejunum and its relation to differentiation of epithelial cells

    Institute of Scientific and Technical Information of China (English)

    Fang-Yu Wang; Masahito Watanabe; Ren-Min Zhu; Kentaro Maemura


    AIM: To investigate the expression between γ-aminobutyric acid (GABA) and glutamate decarboxylase and its relation with differentiation and maturation of jejunal epithelial cells in rat jejunum.METHODS: Immunohistochemical expression of GABA and glutamate decarboxylase (GAD, including two isoforms,GAD65 and GAD67) was investigated in rat jejunum.Meanwhile, double staining was performed with GAD65 immunohistochemistry, followed by lectin histochemistry of fluorescent wheat germ agglutinin. Furthermore,evaluation of cell kinetics in jejunum was conducted by 3Hthymidine autoradiography and immunohistochemistry using a monoclonal antibody to proliferating cell nuclear antigen (PCNA).RESULTS: The cells showing positive immunoreactivity GABA and GAD65 were mainly distributed in the villi in rat jejunum, while jejunal epithelial cells were negative for GAD67. Positive GABA or GAD65 staining was mainly located in the cytoplasm and along the brush border of epithelial cells in the middle and upper portions. In addition, a few GABA and GAD65 strongly positive cells were scattered in the upper two thirds of jejunal villi. Double staining showed that GAD65 immunoreactivity was not found in goblet cells.3H-thymidine-labeled nuclei were found in the lower and middle portions of jejunal crypts, which was consistent with PCNA staining. Therefore, GABA and GAD65 were expressed in a maturation or functional zone.CONCLUSION: The characteristic expression of GABA and GAD suggests that GABA might be involved in regulation of differentiation and maturation of epithelial cells in rat jejunum.

  8. Type I and II positive allosteric modulators differentially modulate agonist-induced up-regulation of α7 nicotinic acetylcholine receptors

    DEFF Research Database (Denmark)

    Thomsen, Morten Skøtt; Mikkelsen, Jens D


    Long-term treatment with nicotine or selective α7 nicotinic acetylcholine receptor (nAChR) agonists increases the number of α7 nAChRs and this up-regulation may be involved in the mechanism underlying the sustained procognitive effect of these compounds. Here, we investigate the influence of type I...... expressing human α7 nAChR, whereas the type I PAMs AVL-3288 or NS1738 do not. Contrarily, neither type I nor II PAMs affect 10 μM nicotine-induced receptor up-regulation, suggesting that nicotine and A-582941 induce up-regulation through different mechanisms. We further show in vivo that 3 mg/kg PNU-120596...... inhibits up-regulation of the α7 nAChR induced by 10 mg/kg A-582941, as measured by [(125)I]-bungarotoxin autoradiography, whereas 1 mg/kg AVL-3288 does not. Given that type II PAMs decrease desensitization of the receptor, whereas type I PAMs do not, these results suggest that receptor desensitization...

  9. Brain-derived neurotrophic factor-deficient mice exhibit a hippocampal hyperserotonergic phenotype. (United States)

    Guiard, Bruno P; David, Denis J P; Deltheil, Thierry; Chenu, Franck; Le Maître, Erwan; Renoir, Thibault; Leroux-Nicollet, Isabelle; Sokoloff, Pierre; Lanfumey, Laurence; Hamon, Michel; Andrews, Anne M; Hen, René; Gardier, Alain M


    Growing evidence supports the involvement of brain-derived neurotrophic factor (BDNF) in mood disorders and the mechanism of action of antidepressant drugs. However, the relationship between BDNF and serotonergic signalling is poorly understood. Heterozygous mutants BDNF +/- mice were utilized to investigate the influence of BDNF on the serotonin (5-HT) system and the activity of the serotonin transporter (SERT) in the hippocampus. The zero net flux method of quantitative microdialysis revealed that BDNF +/- heterozygous mice have increased basal extracellular 5-HT levels in the hippocampus and decreased 5-HT reuptake capacity. In keeping with these results, the selective serotonin reuptake inhibitor paroxetine failed to increase hippocampal extracellular 5-HT levels in BDNF +/- mice while it produced robust effects in wild-type littermates. Using in-vitro autoradiography and synaptosome techniques, we investigated the causes of attenuated 5-HT reuptake in BDNF +/- mice. A significant decrease in [3H]citalopram-binding-site density in the CA3 subregion of the ventral hippocampus and a significant reduction in [3H]5-HT uptake in hippocampal synaptosomes, revealed mainly a decrease in SERT function. However, 5-HT1A autoreceptors were not desensitized in BDNF +/- mice. These results provide evidence that constitutive reductions in BDNF modulate SERT function reuptake in the hippocampus.

  10. Autoradiographic demonstration of oxytocin-binding sites in the macula densa

    Energy Technology Data Exchange (ETDEWEB)

    Stoeckel, M.E.; Freund-Mercier, M.J. (Centre National de la Recherche Scientifique, Strasbourg (France))


    Specific oxytocin (OT)-binding sites were localized in the rat kidney with use of a selective {sup 125}I-labeled OT antagonist ({sup 125}I-OTA). High concentrations of OT binding sites were detected on the juxtaglomerular apparatus with use of the conventional film autoradiographic technique. No labeling occurred on other renal structures. The cellular localization of the OT binding sites within the juxtaglomerular apparatus was studied in light microscope autoradiography, on semithin sections from paraformaldehyde-fixed kidney slices incubated in the presence of {sup 125}I-OTA. These preparations revealed selective labeling of the macula densa, mainly concentrated at the basal pole of the cells. Control experiments showed first that {sup 125}I-OTA binding characteristics were not noticeably altered by prior paraformaldehyde fixation of the kidneys and second that autoradiographic detection of the binding sites was not impaired by histological treatments following binding procedures. In view of the role of the macula densa in the tubuloglomerular feedback, the putative OT receptors of this structure might mediate the stimulatory effect of OT on glomerular filtration.

  11. Correlation between oxytocin neuronal sensitivity and oxytocin receptor binding: An electrophysiological and autoradiographical study comparing rat and guinea pig hippocampus

    Energy Technology Data Exchange (ETDEWEB)

    Raggenbass, M.; Tribollet, E.; Dubois-Dauphin, M.; Dreifuss, J.J. (Univ. Medical Center, Geneva (Switzerland))


    In transverse hippocampal slices from rat and guinea pig brains, the authors obtained unitary extracellular recordings from nonpyramidal neurones located in or near the stratum pyramidale in the CA1 field and in the transition region between the CA1 and the subiculum. In rats, these neurones responded to oxytocin at 50-1,000 nM by a reversible increase in firing rate. The oxytocin-induced excitation was suppressed by a synthetic structural analogue that acts as a potent, selective antioxytocic on peripheral receptors. Nonpyramidal neurones were also excited by carbachol at 0.5-10 {mu}M. The effect of this compound was postsynaptic and was blocked by the muscarinic antagonist atropine. In guinea pigs, by contrast, nonpyramidal neurones were unaffected by oxytocin, although they were excited by carbachol. Light microscopic autoradiography, carried out using a radioiodinated selective antioxytocic as a ligand, revealed labeling in the subiculum and in the CA1 area of the hippocampus of rats, whereas no oxytocin-binding sites were detected in the hippocampus of guinea pigs. The results indicate (i) that a hippocampal action of oxytocin is species-dependent and (ii) that a positive correlation exists between neuronal responsiveness to oxytocin and the presence in the hippocampus of high-affinity binding sites for this peptide.


    Institute of Scientific and Technical Information of China (English)

    P. Wu; D.Y. Yu; X.L. He


    The development of non-equilibrium segregation of boron at grain boundaries in Fe-40%Ni alloy during continuous cooling process was experimentally observed with boronParticle Tracking Autoradiography (PTA) and Transmission Electron Microscopy(TEM). The samples with 10ppm boron were cooled at 2℃/s to 1040, 980, 920,860, 780 and 640℃ respectively after pre-heat treatment of 1150℃ for 15min witha Gleeble-1500 heat simulating machine, then water quenched to room temperature.The width of segregation layer and boron depletion zone, rich factor and other pc-rameters were measured by a special image analysis system. The experimental resultsof PTA show that the grain boundary segregation of boron during cooling process is adynamic process and the development of the non-equilibrium segregation experiencesthree stages: first increases rapidly from 1150 to 1040℃, then gently from 1040 to860℃, and rapidly again from 860℃ to 640℃. The width of boron depletion zoneincreases from about 11μm at 1040℃ to 26μm at 640℃. TEM observation showsthat boron precipitates exist at grain boundaries when the samples are cooled to below860℃. The experimental phenomena are briefly discussed.

  13. Synaptic activation of ribosomal protein S6 phosphorylation occurs locally in activated dendritic domains. (United States)

    Pirbhoy, Patricia Salgado; Farris, Shannon; Steward, Oswald


    Previous studies have shown that induction of long-term potentiation (LTP) induces phosphorylation of ribosomal protein S6 (rpS6) in postsynaptic neurons, but the functional significance of rpS6 phosphorylation is poorly understood. Here, we show that synaptic stimulation that induces perforant path LTP triggers phosphorylation of rpS6 (p-rpS6) locally near active synapses. Using antibodies specific for phosphorylation at different sites (ser235/236 versus ser240/244), we show that strong synaptic activation led to dramatic increases in immunostaining throughout postsynaptic neurons with selectively higher staining for p-ser235/236 in the activated dendritic lamina. Following LTP induction, phosphorylation at ser235/236 was detectable by 5 min, peaked at 30 min, and was maintained for hours. Phosphorylation at both sites was completely blocked by local infusion of the NMDA receptor antagonist, APV. Despite robust induction of p-rpS6 following high frequency stimulation, assessment of protein synthesis by autoradiography revealed no detectable increases. Exploration of a novel environment led to increases in the number of p-rpS6-positive neurons throughout the forebrain in a pattern reminiscent of immediate early gene induction and many individual neurons that were p-rpS6-positive coexpressed Arc protein. Our results constrain hypotheses about the possible role of rpS6 phosphorylation in regulating postsynaptic protein synthesis during induction of synaptic plasticity.

  14. Metabotropic glutamate receptor type 5 in levodopa-induced motor complications. (United States)

    Ouattara, Bazoumana; Grégoire, Laurent; Morissette, Marc; Gasparini, Fabrizio; Vranesic, Ivo; Bilbe, Graeme; Johns, Donald R; Rajput, Alex; Hornykiewicz, Oleh; Rajput, Ali H; Gomez-Mancilla, Baltazar; Di Paolo, Thérèse


    Metabotropic glutamate receptors type 5 (mGluR5) are implicated in regulation of synaptic plasticity and learning, and were the focus of our investigation in human Parkinson's disease (PD) patients with dyskinesias and wearing-off, and in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) monkeys with dyskinesias. Using the selective mGluR5 ligand [(3)H]ABP688 autoradiography, we measured mGluR5 in brain slices from 11 normal and 14 PD patients and from MPTP monkeys, in relation to motor complications (dyskinesias and wearing-off) associated with treatment with l-dopa. In 16 monkeys with a bilateral MPTP lesion and four controls, [(3)H]ABP688 specific binding was elevated in the striatum of dyskinetic l-dopa-treated MPTP monkeys but not in MPTP monkeys without dyskinesias compared to controls. PD patients with motor complications (either dyskinesias or wearing-off) had higher [(3)H]ABP688 specific binding compared to those without motor complications and controls in putamen, external and internal globus pallidus. Elevated glutamatergic transmission as measured with increased mGluR5 specific binding was associated with motor complications and its antagonism could be targeted for their treatment.

  15. Natural radionuclides in the environment. A contribution for the localization and characterization of natural hot particles in solid samples; Natuerliche Radionuklide in der Umwelt. Ein Beitrag zur Lokalisierung und Charakterisierung natuerlicher Heisser Teilchen in festen Proben

    Energy Technology Data Exchange (ETDEWEB)

    Boehnke, A.


    In the present thesis appearance, spreading, origin, and mineralogical properties of natural hot particles are studied and the radioecological relevance of these particles judged. For this first relevent quaestions on radioactivity in the environment and on hot particles are theoretically treated. In the following detailedly the method of the autoradiography and solid-state track detectors is considered, which make possible to quote the precise position, the number and distribution of radioactive particles on the cutting area or surface of a sample. Basing on these methodical considerations by laboratory experiments determined track pattern formations of alpha emitters are documentated and interpreted. Starting from the knowledge obtained from this in the further part of the thesis a detection technique is developed, by means of which it is possible, to determine and mark the position of natural hot particles in sold samples. Thereafter follows a description of the electron-microscopical studies for the identification of the localized natural hot particles. Using the developed detection technique, as well as the electron-microscopical methods, a broad spectrum of samples - anthropogeneous depositions (industrial residues, industry products, by-products) and natural depositions (rocks, sediments, minerals) - is studied.

  16. Visualizing the 17th century underpainting in Portrait of an Old Man by Rembrandt van Rijn using synchrotron-based scanning macro-XRF

    Energy Technology Data Exchange (ETDEWEB)

    Alfeld, Matthias; Janssens, Koen [University of Antwerp, Department of Chemistry, Antwerpen (Belgium); Siddons, D.P. [Brookhaven National Laboratory, National Synchrotron Light Source, Upton, NY (United States); Dik, Joris [Delft University of Technology, Department of Materials Science, Delft (Netherlands); Woll, Arthur [Cornell University, Cornell High Energy Synchrotron Source, Ithaca, NY (United States); Kirkham, Robin [CSIRO, Materials Science and Engineering, Clayton, VIC (Australia); Wetering, Ernst van de [Rembrandt Research Project, Amsterdam (Netherlands)


    In 17th century Old Master Paintings, the underpainting generally refers to the first sketch of a composition. The underpainting is applied to a prepared ground using a monochrome, brown oil paint to roughly indicate light, shade and contours. So far, methods to visualize the underpainting - other than in localized cross-sections - have been very limited. Neither infrared reflectography nor neutron induced autoradiography have proven to be practical, adequate visualization tools. Thus, although of fundamental interest in the understanding of a painting's genesis, the underpainting has virtually escaped all imaging efforts. In this contribution we will show that 17th century underpainting may consist of a highly heterogeneous mixture of pigments, including copper pigments. We suggest that this brown pigment mixture is actually the recycled left-over of a palette scraping. With copper as the heaviest exclusive elemental component, we will hence show in a case study on a Portrait of an Old Man attributed to Rembrandt van Rijn how scanning macro-XRF can be used to efficiently visualize the underpainting below the surface painting and how this information can contribute to the discussion of the painting's authenticity. (orig.)

  17. PCP-induced alterations in cerebral glucose utilization in rat brain: blockade by metaphit, a PCP-receptor-acylating agent

    Energy Technology Data Exchange (ETDEWEB)

    Tamminga, C.A.; Tanimoto, K.; Kuo, S.; Chase, T.N.; Contreras, P.C.; Rice, K.C.; Jackson, A.E.; O' Donohue, T.L.


    The effects of phencyclidine (PCP) on regional cerebral glucose utilization was determined by using quantitative autoradiography with (/sup 14/C)-2-deoxyglucose. PCP increased brain metabolism in selected areas of cortex, particularly limbic, and in the basal ganglia and thalamus, whereas the drug decreased metabolism in areas related to audition. These results are consistent with the known physiology of central PCP neurons and may help to suggest brain areas involved in PCP-mediated actions. Moreover, based on the behavioral similarities between PCP psychosis and an acute schizophrenic episode, these data may be relevant to the understanding of schizophrenia. The PCP-receptor-acylating agent, metaphit, blocked most of these PCP actions. In addition, metaphit by itself was found to diminish glucose utilization rather uniformly throughout brain. These results indicate an antagonist effect of metaphit on the PCP system and suggest a widespread action of metaphit, putatively at a PCP-related site, possibly in connection with the N-methyl-D-aspartate (NMDA) receptor.

  18. Neurogenesis in the brain auditory pathway of a marsupial, the northern native cat (Dasyurus hallucatus)

    Energy Technology Data Exchange (ETDEWEB)

    Aitkin, L.; Nelson, J.; Farrington, M.; Swann, S. (Department of Physiology, Monash University, Melbourne (Australia))


    Neurogenesis in the auditory pathway of the marsupial Dasyurus hallucatus was studied. Intraperitoneal injections of tritiated thymidine (20-40 microCi) were made into pouch-young varying from 1 to 56 days pouch-life. Animals were killed as adults and brain sections were prepared for autoradiography and counterstained with a Nissl stain. Neurons in the ventral cochlear nucleus were generated prior to 3 days pouch-life, in the superior olive at 5-7 days, and in the dorsal cochlear nucleus over a prolonged period. Inferior collicular neurogenesis lagged behind that in the medial geniculate, the latter taking place between days 3 and 9 and the former between days 7 and 22. Neurogenesis began in the auditory cortex on day 9 and was completed by about day 42. Thus neurogenesis was complete in the medullary auditory nuclei before that in the midbrain commenced, and in the medial geniculate before that in the auditory cortex commenced. The time course of neurogenesis in the auditory pathway of the native cat was very similar to that in another marsupial, the brushtail possum. For both, neurogenesis occurred earlier than in eutherian mammals of a similar size but was more protracted.

  19. Neurogenesis in Aplysia californica resembles nervous system formation in vertebrates. [Sponges

    Energy Technology Data Exchange (ETDEWEB)

    Jacob, M.H.


    The pattern of neurogenesis of the central nervous system of Aplysia californica was investigated by (/sup 3/H)thymidine autoradiography. Large numbers of animals at a series of early developmental stages were labeled with (/sup 3/H)thymidine for 24 or 48 hr and were subsequently sampled at specific intervals throughout the life cycle. I found that proliferative zones, consisting of columnar and placodal ectodermal cells, are established in regions of the body wall adjacent to underlying mesodermal cells. Mitosis in the proliferative zones generates a population of cells which leave the surface and migrate inward to join the nearby forming ganglia. Tracing specific (/sup 3/H)thymidine-labeled cells from the body wall to a particular ganglion and within the ganglion over time suggests that the final genomic replication of the neuronal precursors occurs before the cells join the ganglion while glial cell precursors and differentiating glial cells continue to divide within the ganglion for some time. Ultrastructural examination of the morphological features of the few mitosing cells observed within the Aplysia central nervous system supports this interpretation. The pattern of neurogenesis in the Aplysia central nervous system resembles the proliferation of cells in the neural tube and the migration of neural crest and ectodermal placode cells in the vertebrate nervous system but differs from the pattern described for other invertebrates.

  20. Protein turnover and cellular autophagy in growing and growth-inhibited 3T3 cells

    Energy Technology Data Exchange (ETDEWEB)

    Papadopoulos, T.; Pfeifer, U. (Univ. of Wuerzburg (West Germany))


    The relationship between growth, protein degradation, and cellular autophagy was tested in growing and in growth-inhibited 3T3 cell monolayers. For the biochemical evaluation of DNA and protein metabolism, growth-inhibited 3T3 cell monolayers with high cell density and growing 3T3 cell monolayers with low cell density were labeled simultaneously with ({sup 14}C)thymidine and ({sup 3}H)leucine. The evaluation of the DNA turnover and additional ({sup 3}H)thymidine autoradiography showed that 24 to 5% of 3T3 cells continue to replicate even in the growth-inhibited state, where no accumulation of protein and DNA can be observed. Cell loss, therefore, has to be assumed to compensate for the ongoing cell proliferation. When the data of protein turnover were corrected for cell loss, it was found that the rate constant of protein synthesis in nongrowing monolayers was reduced to half the value found in growing monolayers. Simultaneously, the rate constant of protein degradation in nongrowing monolayers was increased to about 1.5-fold the value of growing monolayers. These data are in agreement with the assumption that cellular autophagy represents a major pathway of regulating protein degradation in 3T3 cells and that the regulation of autophagic protein degradation is of relevance for the transition from a growing to a nongrowing state.

  1. Functionalization of Mesoporous Silicon Nanoparticles for Targeting and Bioimaging Purposes

    Directory of Open Access Journals (Sweden)

    Jussi Rytkönen


    Full Text Available Carboxylic acid functionalized thermally hydrocarbonized porous silicon nanoparticles (UnTHCPSi-NP were synthesized, and their opsonization and targeting properties were studied in vitro alongside with in vivo biodistribution. The carboxyl groups on UnTHCPSi were utilized to further functionalize the nanoparticles. In order to reduce the opsonization of the UnTHCPSi-NPs, different sized polyethylene glycol (PEG were conjugated or adsorbed to the NPs surface. The latter approach, based on hydrophobic interaction, turned out to be more effective in reducing the opsonization and improving the stability of the nanoparticle suspension. The most abundant opsonins after plasma incubation were fibrinogen precursors and IgG. Furthermore, the targeting properties of UnTHCPSi-NPs were studied in vitro with antibodies against glutathione S-transferase (anti-GST. PEGylated NPs conjugated with anti-GST bound to GST-agarose in human plasma nearly 35-fold compared to control NPs, indicating that UnTHCPSi-NPs are suitable for targeting in physiological environment. The in vivo biodistribution in mice revealed that PEGylated UnTHCPSi-NPs, accumulate fast into the liver and the spleen, regardless of the reduced opsonization in vitro. However, autoradiography and transmission electron microscopy showed that majority of the NPs still remained in hepatic blood vessels and sinusoids suggesting a possibility to utilize them as a sustained release platform for payload molecules.

  2. Localization of cannabinoid CB1 receptor mRNA using ribonucleotide probes: methods for double- and single-label in situ hybridization. (United States)

    Hohmann, Andrea G


    This chapter presents a reliable, detailed method for performing double-label in situ hybridization (ISH) that has been validated for use in studies identifying the co-localization of cannabinoid CB1 receptor mRNA with other distinct species of mRNAs. This method permits simultaneous detection of two different species of mRNA within the same tissue section. Double-label ISH may be accomplished by hybridizing tissue sections with a combination of radiolabeled and digoxigenin-labeled RNA probes that are complementary to their target mRNAs. Single-label ISH may be accomplished by following the procedures described for use with radioisotopic probes (here [35S]-labeled) only. Silver grains derived from conventional emulsion autoradiography are used to detect the radiolabeled cRNA probe. An alkaline phosphatase-dependent chromogen reaction product is used to detect the nonisotopic (here, digoxigenin-labeled) cRNA probe. Necessary controls that are required to document the specificity of the labeling of the digoxigenin and radiolabeled probes are described. The methods detailed herein may be employed to detect even low levels of a target mRNA. These methods may be utilized to study co-localization and coregulation of expression of a particular gene within identified neurons in multiple systems.

  3. Carbon-11 labeled stilbene derivatives from natural products for the imaging of Aβ plaques in the brain

    Energy Technology Data Exchange (ETDEWEB)

    Cui, Mengchao; Tang, Ruikun; Li, Zijing; Jia, Hongmei; Liu, Boli [Beijing Normal Univ. (China). Key Laboratory of Radiopharmaceuticals; Zhang, Jinming; Zhang, Xiaojun [Chinese PLA General Hospital, Beijing (China). Dept. of Nuclear Medicine


    Four stilbene derivatives from natural products were screened as novel β-amyloid (Aβ) imaging ligands. In vitro binding assay showed that the methylated ligand, (E)-1-methoxy-4-styrylbenzene (8) displayed high binding affinity to Aβ{sub 1-42} aggregates (K{sub i} = 19.5 nM). Moreover, the {sup 11}C-labeled ligand, [{sup 11}C]8 was prepared through an O-methylation reaction using [{sup 11}C]CH{sub 3}OTf. In vitro autoradiography with sections of transgenic mouse brain also confirmed the high and specific binding of [{sup 11}C]8 to Aβ plaques. In vivo biodistribution experiments in normal mice indicated that [{sup 11}C]8 displayed high initial uptake (9.41 ± 0.51% ID/g at 5 min post-injection) into and rapid washout from the brain, with a brain{sub 5} {sub min}/brain{sub 30} {sub min} ratio of 6.63. These preliminary results suggest that [{sup 11}C]8 may be served as a novel Aβ imaging probe for PET. (orig.)

  4. Anomalous Nuclear Phenomena Associated with Ultrafast Processes (United States)

    Jiang, Xingliu; Zhou, Xiaoping; Han, Lijun; Wang, Liyin


    Localized nuclear reactions on the tips of the surface of electrodes in electrolysis cells have been observed by using solid detectors CR-39 and autoradiography in our laboratory at the period of May, 1989. A physical model of transient vortex dynamics with torsion coherence with the zero point energy has been proposed by Xingliu Jiang based on the ultrafast processes of tripple phases area of tip effect on the electrode surface. Considering the large equivelent capacitance of electrochemical double layer, it is presumed that the double layer can exhibit nonlinear electrical response with spatial and temporal variations confined to micreoscopic areas by tip effect. Recent work reveals that nuclear reactions which usually occur at the field of high energy states, could be created in the systems of far from equilibrium with nonlinear beharvior at room tempurature.Our current understanging of science is like a puzzle with a large missing piece-zero point energy. Jiang Xingliu, Lei Jinzhi, Torsion field and tapping the zero-point energy in an electrochemical system, J. of New Energy, 4(2), 93(1999). B. Naranjo, J.K. Gimzewski & S. Putterman, Observation of nuclear fusion driven by a pyroelectric crystal, Nature, 434, 1115(2005).

  5. Measurement of specific parameters for dose calculation after inhalation of aerols containing transuranium elements; Mesure de parametres specifiques pour le calcul de dose apres inhalation d'aerosols renfermant des elements transuraniens

    Energy Technology Data Exchange (ETDEWEB)

    Ramounet-le Gall, B.; Fritsch, P.; Abram, M.C.; Rateau, G.; Grillon, G.; Guillet, K. [Lab. de Radiotoxicologie, CEA/DSV/DRR/SRCA, Bruyeres le Chatel (France); Baude, S. [Lab. de Mesures Specifiques Gaz, CEA/DAM/DASE/SRCE, Bruyeres le Chatel (France); Berard, P. [Cabinet du Conseiller medical du CEA, CEA/DEN/DPS/LABM Saclay, Gif sur Yvette (France); Ansoborlo, E. [CEA/DEN/DRCP/CETAMA, Bagnols sur Ceze (France); Delforge, J. [Lab. de Radiotoxicologie, CEA/DSV/DRR/SRCA, Bruyeres le Chatel (France)


    A review on specific parameter measurements to calculate doses per unit of incorporation according to recommendations of the International Commission of Radiological Protection has been performed for inhaled actinide oxides. Alpha activity distribution of the particles can be obtained by autoradiography analysis using aerosol sampling filters at the work places. This allows us to characterize granulometric parameters of 'pure' actinide oxides, but complementary analysis by scanning electron microscopy is needed for complex aerosols. Dissolution parameters with their standard deviation are obtained after rat inhalation exposure, taking into account both mechanical lung clearance and actinide transfer to the blood estimated from bone retention. In vitro experiments suggest that the slow dissolution rate might decrease as a function of time following exposure. Dose calculation software packages have been developed to take into account granulometry and dissolution parameters as well as specific physiological parameters of exposed individuals. In the case of poorly soluble actinide oxides, granulometry and physiology appear as the main parameters controlling dose value, whereas dissolution only alters dose distribution. Validation of these software packages are in progress. (author)

  6. Dopamine D₄ receptor counteracts morphine-induced changes in µ opioid receptor signaling in the striosomes of the rat caudate putamen. (United States)

    Suárez-Boomgaard, Diana; Gago, Belén; Valderrama-Carvajal, Alejandra; Roales-Buján, Ruth; Van Craenenbroeck, Kathleen; Duchou, Jolien; Borroto-Escuela, Dasiel O; Medina-Luque, José; de la Calle, Adelaida; Fuxe, Kjell; Rivera, Alicia


    The mu opioid receptor (MOR) is critical in mediating morphine analgesia. However, prolonged exposure to morphine induces adaptive changes in this receptor leading to the development of tolerance and addiction. In the present work we have studied whether the continuous administration of morphine induces changes in MOR protein levels, its pharmacological profile, and MOR-mediated G-protein activation in the striosomal compartment of the rat CPu, by using immunohistochemistry and receptor and DAMGO-stimulated [35S]GTPγS autoradiography. MOR immunoreactivity, agonist binding density and its coupling to G proteins are up-regulated in the striosomes by continuous morphine treatment in the absence of changes in enkephalin and dynorphin mRNA levels. In addition, co-treatment of morphine with the dopamine D4 receptor (D4R) agonist PD168,077 fully counteracts these adaptive changes in MOR, in spite of the fact that continuous PD168,077 treatment increases the [3H]DAMGO Bmax values to the same degree as seen after continuous morphine treatment. Thus, in spite of the fact that both receptors can be coupled to Gi/0 protein, the present results give support for the existence of antagonistic functional D4R-MOR receptor-receptor interactions in the adaptive changes occurring in MOR of striosomes on continuous administration of morphine.

  7. Nimodipine pretreatment improves cerebral blood flow and reduces brain edema in conscious rats subjected to focal cerebral ischemia. (United States)

    Jacewicz, M; Brint, S; Tanabe, J; Wang, X J; Pulsinelli, W A


    The effect of nimodipine pretreatment on CBF and brain edema was studied in conscious rats subjected to 2.5 h of focal cortical ischemia. An infusion of nimodipine (2 micrograms/kg/min i.v.) or its vehicle, polyethylene glycol 400, was begun 2 h before the ischemic interval and was continued throughout the survival period. Under brief halothane anesthesia, the animals' right middle cerebral and common carotid arteries were permanently occluded, and 2.5 h later, they underwent a quantitative CBF study ([14C]iodoantipyrine autoradiography followed by Quantimet 970 image analysis). Nimodipine treatment improved blood flow to the middle cerebral artery territory without evidence of a "vascular steal" and reduced the volume of the ischemic core (cortex with CBF of less than 25 ml/100 g/min) and accompanying edema by approximately 50% when compared with controls (p = 0.006 and 0.0004, respectively). Mild hypotension induced by nimodipine did not aggravate the ischemic insult. The ischemic core volumes, however, were 50-75% smaller than the 24-h infarct volumes generated in a similar paradigm that demonstrated 20-30% infarct reduction with continuous nimodipine treatment. These results suggest that nimodipine pretreatment attenuates the severity of early focal cerebral ischemia, but that with persistent ischemia, cortex surrounding the ischemic core undergoes progressive infarction and the early benefit of nimodipine treatment is only partly preserved.

  8. Posthoc phosphorylation of proteins derived from ischemic rat hippocampus, striatum and neocortex. (United States)

    Kirschenbaum, B; Pulsinelli, W A


    Disruption of the brain's protein phosphorylation system by ischemia may cause irreversible metabolic and structural alterations leading eventually to cell death. To examine the effect of ischemia on the phosphorylation state of brain proteins, tissue homogenates derived from the hippocampus, striatum and neocortex of normal rats and rats subjected to severe forebrain ischemia were phosphorylated with [gamma-32P]ATP. The phosphorylated proteins were separated by two-dimensional polyacrylamide gel electrophoresis and changes were assessed by autoradiography. Cerebral ischemia caused marked alterations of the phosphorylation state of many brain proteins; phosphorylation of some proteins was increased while phosphorylation of others was decreased. Despite differences in the sensitivity of the hippocampus, striatum and neocortex to ischemic injury the direction and approximate magnitude of protein phosphorylation changes caused by ischemia were similar in all three regions. Since the pattern of protein phosphorylation in the ischemia-vulnerable hippocampus was identical to that in the ischemia-resistant paramedian neocortex we conclude that abnormalities of protein phosphorylation may be necessary for ischemic injury to neurons but none are sufficient to explain the selective vulnerability of certain brain regions to ischemic damage.

  9. A 55,000-60,000 Mr receptor protein for urokinase-type plasminogen activator. Identification in human tumor cell lines and partial purification

    DEFF Research Database (Denmark)

    Nielsen, L S; Kellerman, G M; Behrendt, N;


    The iodinated Mr approximately equal to 15,000 amino-terminal fragment (ATF) of the urokinase-type plasminogen activator (u-PA) molecule bound specifically to the cell surface of all of seven cultured human tumor cell lines studied. Cross-linking of iodinated ATF to the cell surface using...... a bifunctional amino-reactive reagent followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography revealed with the four cell lines studied the occurrence of a single band migrating with an Mr of 70,000-75,000, indicating complex formation with an Mr of 55,000-60,000 u-PA receptor......-PA-R consists of one polypeptide chain. Two forms of u-PA-R, which differed with respect to affinity to concanavalin A, were identified. u-PA-R retained its ability to bind to ATF after cell lysis, and it was purified approximately 2,200-fold from biosynthetically labeled U937 cells by affinity chromatography...

  10. In vivo characterization of radioiodinated (+)-2-[4-(4-iodophenyl) piperidino] cyclohexanol as a potential {sigma}-1 receptor imaging agent

    Energy Technology Data Exchange (ETDEWEB)

    Akhter, Nasima [Department of Biotracer Medicine, Kanazawa University, Kanazawa 920-8640 (Japan); Shiba, Kazuhiro [Division of Tracer Kinetics, Advanced Science Research Center, Kanazawa University, Kanazawa 920-8640 (Japan)], E-mail:; Ogawa, Kazuma [Division of Tracer Kinetics, Advanced Science Research Center, Kanazawa University, Kanazawa 920-8640 (Japan); Kinuya, Seigo; Nakajima, Kenichi [Department of Biotracer Medicine, Kanazawa University, Kanazawa 920-8640 (Japan); Mori, Hirofumi [Division of Tracer Kinetics, Advanced Science Research Center, Kanazawa University, Kanazawa 920-8640 (Japan)


    In this study, the (+)-enantiomer of radioiodinated 2-[4-(4-iodophenyl)piperidino]cyclohexanol [(+)-[{sup 125}I]-p-iodovesamicol] [(+)-[{sup 125}I]pIV], which is reported to bind with high affinity to {sigma}-1 receptors in vitro, was tested for its usefulness in imaging {sigma}-1 receptors in the central nervous system (CNS) in vivo. In biodistribution studies, significant amounts (approximately 3% of the injected dose) of (+)-[{sup 125}I]pIV accumulated in rat brain, and its retention was prolonged. In blocking studies, the accumulation of (+)-[{sup 125}I]pIV in the rat brain was significantly reduced by the coadministration of {sigma}-ligands such as pentazocine (5.0 {mu}mol), haloperidol (0.5 {mu}mol) or SA4503 (0.5 {mu}mol). The blocking effect of pentazocine (selective {sigma}-1 ligand) was similar to the blocking effects of SA4503 and haloperidol [nonselective {sigma} ({sigma}-1 and {sigma}-2) ligands]. Ex vivo autoradiography of the rat brain at 45 min following intravenous injection of (+)-[{sup 125}I]pIV showed high localization in brain areas rich in {sigma}-1 receptors. Thus, the distribution of (+)-[{sup 125}I]pIV was thought to bind to {sigma}-1 receptors in the CNS in vivo. These results indicate that radioiodinated (+)-pIV may have the potential to image {sigma}-1 receptors in vivo.

  11. Radioactive labeling of a natural assemblage of marine sedimentary bacteria and microalgae for trophic studies: An autoradiographic study. (United States)

    Carman, K R


    Autoradiography was used to examine critical questions for trophic studies concerning the uptake of radioactive tracers by a natural assemblage of sedimentary microorganisms. Labeled organic substrates ([(3)H]-acetate and [(3)H]-thymidine) were taken up only by heterotrophic bacteria, and [(14)C]-bicarbonate was taken up only by microalgae. Only approximately 2% of the bacterial assemblage took up detectable quantities of either [(3)H]-acetate or [(3)H]-thymidine, regardless of whether labeled substrates were delivered to sediments via slurries or by injection with a microliter syringe. Significantly more diatoms were labeled when [(14)C]-bicarbonate was delivered to sediments by the injection method (75%) as compared to the slurry method (50%). These results indicate that radio-active tracers can be used in natural sediments to selectively label potential microbial food of invertebrate grazers. Only a small proportion of bacteria, however, may actually use a labeled substrate, which introduces a large uncertainty into the conversion of radioactivity in grazers to the number of bacteria consumed. Finally, the use of disruptive methods (e.g., slurries) to deliver labels to sediments does not increase the proportion of microorganisms that become labeled. Thus, given the variety of artifacts that may be associated with the use of sediment slurries, it is probably advisable to use nondisruptive methods to deliver substrates to sediments.

  12. The distribution of radioactivity in brains of rats given (N-methyl- sup 11 C)PK 11195 in vivo after induction of a cortical ischaemic lesion

    Energy Technology Data Exchange (ETDEWEB)

    Cremer, J.E.; Hume, S.P.; Cullen, B.M.; Myers, R.; Manjil, L.G.; Turton, D.R.; Luthra, S.K.; Bateman, D.M.; Pike, V.W. (Hammersmith Hospital, London (United Kingdom). M.R.C. Cyclotron Unit)


    PK 11195 is a selective ligand for the peripheral-type benzodiazepine bindings site (PTBBS). There are few sites in normal brain but their number increases in association with tissue necrosis. The time-course of appearance of PTBBS around a focally induced ischaemic lesion in frontal cortex of rat brain was established by autoradiography using (N-methyl-{sup 3} H )PK 11195. Using this information and the same experimental model of ischaemia, the distribution of radioactivity after injection of carbon-11 labelled PK 11195 was studied. The purpose was to synthesize (N-methyl-{sup 11}C)PK 11195 and to test its suitability as a tracer for depicting the presence of PTBB in ischaemic lesions. The time-profiles of distribution of radioactivity in brain regions after intravenous injection of tracer and the ratio of radioactivity in lesioned compared with unlesioned cortex were determined. Data for the temporal (days after lesion induction) and for the regional retention of radioactivity were consistent with independent evidence (autoradiographic and immunohistochemical) for the occurence of increased numbers of PTBBS, predominantly in association with macrophages, in areas undergoing necrosis. (Author).

  13. Novel Fluorine-Containing NMDA Antagonists for Brain Imaging: In Vitro Evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Alvarado, M.; Biegon, A.


    The NMDA receptor has been implicated in neuronal death following stroke, brain injury and neurodegenerative disorders (e.g. Alzheimer's, Parkinson's and Huntington's disease) and in physiological functions (e.g. memory and cognition). Non-competitive antagonists, such as MK- 801 and CNS-1102, that block the action of glutamate at the NMDA receptor have been shown to be neuroprotective by blocking the influx of calcium into the cells. As a result, they are being considered as therapeutic agents for the above mentioned diseases. Several Fluorine-containing novel analogs of NMDA channel blockers have been synthesized and evaluated in search of a compound suitable for 18F labeling and Positron Emission Tomography (PET). Based on in vitro binding assay studies on rat brain membranes, the novel compounds examined displayed a range of affinities. Preliminary analyses indicated that chlorine is the best halogen on the ring, and that ethyl fluoro derivatives are more potent than methyl-fluoro compounds. Further analysis based on autoradiography will be needed to examine the regional binding characteristics of the novel compounds examined in this study. Labeling with 18F will allow the use of these compounds in humans, generating new insights into mechanisms and treatment of diseases involving malfunction of the glutamatergic system in the brain.

  14. In vivo radiometric analysis of glucose uptake and distribution in mouse bone

    Institute of Scientific and Technical Information of China (English)

    Meredith L Zoch; Diane S Abou; Thomas L Clemens; Daniel LJ Thorek; and Ryan C Riddle


    Bone formation and remodeling occurs throughout life and requires the sustained activity of osteoblasts and osteoclasts, particularly during periods of rapid bone growth. Despite increasing evidence linking bone cell activity to global energy homeostasis, little is known about the relative energy requirements or substrate utilization of bone cells. In these studies, we measured the uptake and distribution of glucose in the skeleton in vivo using positron-emitting 18F-fluorodeoxyglucose ([18F]-FDG) and non-invasive, high-resolution positron emission tomography/computed tomography (PET/CT) imaging and ex vivo autoradiography. Assessment of [18F]-FDG uptake demonstrated that relative to other tissues bone accumulated a significant fraction of the total dose of the glucose analog. Skeletal accumulation was greatest in young mice undergoing the rapid bone formation that characterizes early development. PET/CT imaging revealed that [18F]-FDG uptake was greatest in the epiphyseal and metaphyseal regions of long bones, which accords with the increased osteoblast numbers and activity at this skeletal site. Insulin administration significantly increased skeletal accumulation of [18F]-FDG, while uptake was reduced in mice lacking the insulin receptor specifically in osteoblasts or fed a high-fat diet. Our results indicated that the skeleton is a site of significant glucose uptake and that its consumption by bone cells is subject to regulation by insulin and disturbances in whole-body metabolism.

  15. Effect of MK-801 on methamphetamine-induced dopaminergic neurotoxicity: long-term attenuation of methamphetamine-induced dopamine release

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Sang Eun; Kim, Yu Ri; Hwang, Se Hwan [Sungkyunkwan Univ., School of Medicine, Seoul (Korea, Republic of)


    Repeated administration of methamphetamine (METH) produces high extracellular levels of dopamine (DA) and subsequent striatal DA terminal damage. The effect of MK-801, a noncompetitive N-methyl-D-aspartate receptor antagonist, on METH-induced changes in DA transporter (DAT) and DA release evoked by an acute METH challenge was evaluated in rodent striatum using [{sup 3}H] WIN 38,428 ex vivo auto-radiography and in vivo microdialysis. Four injections of METH (10 mg/kg, i.p.), each given 2 h apart, produced 71% decrease in DAT levels in mouse striatum 3 d after administration. Pretreatment with MK-801 (2.5 g/kg, i.p.) 15 min before each of the four METH injections protected completely against striatal DAT depletions. Four injections of MK-801 alone did not significantly change striatal DAT levels. Striatal DA release evoked by an acute METH challenge (4mg/kg, i.p.) at 3 d after repeated administration of METH in rats was decreased but significant compared with controls, which was attenuated by repeated pretreatment with MK-801. Also, repeated injections of MK-801 alone attenuated acute METH-induced striatal DA release 3 d after administration. These results suggest that repeated administration of MK-801 may exert a preventive effect against METH-induced DA terminal injury through long-term attenuation of DA release induced by METH and other stimuli.

  16. AMPA receptor potentiation can prevent ethanol-induced intoxication. (United States)

    Jones, Nicholas; Messenger, Marcus J; O'Neill, Michael J; Oldershaw, Anna; Gilmour, Gary; Simmons, Rosa M A; Iyengar, Smriti; Libri, Vincenzo; Tricklebank, Mark; Williams, Steve C R


    We present a substantial series of behavioral and imaging experiments, which demonstrate, for the first time, that increasing AMPA receptor-mediated neurotransmission via administration of potent and selective biarylsulfonamide AMPA potentiators LY404187 and LY451395 reverses the central effects of an acutely intoxicating dose of ethanol in the rat. Using pharmacological magnetic resonance imaging (phMRI), we observed that LY404187 attenuated ethanol-induced reductions in blood oxygenation level dependent (BOLD) in the anesthetized rat brain. A similar attenuation was apparent when measuring local cerebral glucose utilization (LCGU) via C14-2-deoxyglucose autoradiography in freely moving conscious rats. Both LY404187 and LY451395 significantly and dose-dependently reversed ethanol-induced deficits in both motor coordination and disruptions in an operant task where animals were trained to press a lever for food reward. Both prophylactic and acute intervention treatment with LY404187 reversed ethanol-induced deficits in motor coordination. Given that LY451395 and related AMPA receptor potentiators/ampakines are tolerated in both healthy volunteers and elderly patients, these data suggest that such compounds may form a potential management strategy for acute alcohol intoxication.

  17. Nitro-fatty acid pharmacokinetics in the adipose tissue compartment. (United States)

    Fazzari, Marco; Khoo, Nicholas K H; Woodcock, Steven R; Jorkasky, Diane K; Li, Lihua; Schopfer, Francisco J; Freeman, Bruce A


    Electrophilic nitro-FAs (NO2-FAs) promote adaptive and anti-inflammatory cell signaling responses as a result of an electrophilic character that supports posttranslational protein modifications. A unique pharmacokinetic profile is expected for NO2-FAs because of an ability to undergo reversible reactions including Michael addition with cysteine-containing proteins and esterification into complex lipids. Herein, we report via quantitative whole-body autoradiography analysis of rats gavaged with radiolabeled 10-nitro-[(14)C]oleic acid, preferential accumulation in adipose tissue over 2 weeks. To better define the metabolism and incorporation of NO2-FAs and their metabolites in adipose tissue lipids, adipocyte cultures were supplemented with 10-nitro-oleic acid (10-NO2-OA), nitro-stearic acid, nitro-conjugated linoleic acid, and nitro-linolenic acid. Then, quantitative HPLC-MS/MS analysis was performed on adipocyte neutral and polar lipid fractions, both before and after acid hydrolysis of esterified FAs. NO2-FAs preferentially incorporated in monoacyl- and diacylglycerides, while reduced metabolites were highly enriched in triacylglycerides. This differential distribution profile was confirmed in vivo in the adipose tissue of NO2-OA-treated mice. This pattern of NO2-FA deposition lends new insight into the unique pharmacokinetics and pharmacologic actions that could be expected for this chemically-reactive class of endogenous signaling mediators and synthetic drug candidates.

  18. Imaging plasma docosahexaenoic acid (dha incorporation into the brain in vivo, as a biomarker of brain DHA: Metabolism and neurotransmission

    Directory of Open Access Journals (Sweden)

    Rapoport Stanley I.


    Full Text Available Docosahexaenoic acid (DHA is critical for normal brain structure and function, and its brain concentration depends on dietary DHA content and hepatic conversion from its dietary derived n-3 precursor, a-linolenic acid (α-LNA. We developed an in vivo method in rats using quantitative autoradiography to image incorporation into brain of unesterified plasma DHA, and showed that the incorporation rate equals the rate of brain metabolic DHA consumption. Thus, quantitative imaging of DHA incorporation from plasma into brain can be used as a biomarker of brain DHA metabolism and neurotransmission. The method has been extended to humans with the use of positron emission tomography (PET. Furthermore, imaging in unanesthetized rats using DHA incorporation as a biomarker in response to N-methyl-D-aspartate (NMDA administration confirms that regional DHA signaling is independent of extracellular calcium, and likely mediated by a calcium-independent phospholipase A2 (iPLA2. Studies in mice in which iPLA2-VIA (β was knocked out confirmed that this enzyme is critical for baseline and muscarinic cholinergic signaling involving DHA.

  19. Light-dependent degradation of the QB-protein in isolated pea thylakoids (United States)

    Ohad, I.; Kyle, D. J.; Hirschberg, J.


    The 32 000-dalton QB-protein of photosystem II (PS II) is rapidly damaged and removed from isolated pea thylakoids during incubation in the light resulting in a loss of photosynthetic electron flow through PS II. This in vitro photoinhibition is similar to that previously reported with intact Chlamydomonas cells. The damage occurs at a faster rate in vitro, however, due to the inability of isolated thylakoids to synthesize replacement QB-protein. The removal of the damaged QB-protein does not require any soluble components of the chloroplast stroma and is unaffected by the protease inhibitors phenyl-methylsulfonylfluoride or antipain. Unlike the effect of trypsin, no low mol. wt. membrane-bound or soluble fragments of the labelled QB-protein could be identified either by autoradiography or immunologically using polyclonal antibodies specific for the QB-protein. The lightinduced damage to the QB-protein (indicated by a loss of QB functional activity), preceded the removal of the protein from the membrane. We conclude that photodamage of the QB-protein generates a conformational change which renders the protein susceptible to attack by a highly efficient, intrinsic membrane protease. ImagesFig. 1.Fig. 3.Fig. 4.Fig. 5. PMID:16453621

  20. Mammary excretion and placental transfer of cis-malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolane] platinum(II) in rats. (United States)

    Cho, Y B; Kim, K H; Kim, D K; Miyamoto, G


    The mammary excretion and placental transfer of cis-malonato[(4R,5R)-4,5-bis(aminomethyl)-2-isopropyl-1,3-dioxolan e] platinum(II) (CAS 146665-77-2, SKI 2053R), a new potential anticancer agent, were investigated in the lactating or pregnant rats after a single intravenous administration of 14C-SKI 2053R (20 mg/kg, 100 microCi/kg). The radioactivity of the milk declined in a biexponential fashion with an initial half-life of 0.39 h and with a terminal half-life of 14.05 h in the lactating rats. The radioactivity of the milk was lower than that of plasma until 1 h after dosing, but was higher than that of plasma from 4 h after dosing. 14C-SKI 2053R was well distributed to most tissues including uterus and placenta in the pregnant rats, but the levels of radioactivity in the amniotic fluid and fetuses were markedly lower than that in the maternal plasma. Therefore, it is concluded that SKI 2053R scarcely passes the blood-placenta barrier, which was confirmed by the whole-body autoradiography study.

  1. Autoradiographic localization of beta-adrenoreceptors in rat uterus

    Energy Technology Data Exchange (ETDEWEB)

    Tolszczuk, M.; Pelletier, G.


    The inhibitory effects of catecholamines on uterine smooth muscle are known to be mediated through beta-adrenergic receptors. To investigate further the distribution of these receptors in the rat uterus, we utilized in vitro autoradiography using ( SVI)-cyanopindolol (CYP), a specific beta-receptor ligand that has equal activity for both beta 1- and beta 2-receptor subtypes. The specificity of the labeling and the characterization of receptor subtypes in different cell types were achieved by displacement of radioligand with increasing concentrations of zinterol, a beta-adrenergic agonist with preferential affinity for the beta 2-adrenoreceptor subtype, and practolol, a beta-adrenergic antagonist that binds preferentially to the beta 1-subtype. Quantitative estimation of ligand binding was performed by densitometry. It was shown that the vast majority of beta-adrenoreceptors were of the beta 2-subtype and were found in high concentration not only in the myometrium but also in the endometrial and serosal epithelia. Specific labeling was also observed in glandular elements. These results suggest that beta-adrenoreceptors might be involved in different functions in the uterus.

  2. Effect of misoprostol and cimetidine on gastric cell labeling index

    Energy Technology Data Exchange (ETDEWEB)

    Fich, A.; Arber, N.; Sestieri, M.; Zajicek, G.; Rachmilewitz, D.


    The effect of misoprostol and cimetidine on gastric cell turnover was studied. Endoscopic biopsy specimens of fundic and antral mucosa were obtained from duodenal ulcer patients before and after 4 wk of therapy with cimetidine 1.2 g/day or misoprostol 800 micrograms/day. Biopsy specimens were incubated with (/sup 3/H)thymidine. Glandular column length and number of labeled cells were determined after autoradiography. There was no significant difference in column length of antral or fundic glands before or after therapy with cimetidine and misoprostol. The number of antral and fundic labeled cells was significantly decreased after misoprostol treatment (3.6 +/- 0.3 and 4.6 +/- 0.4, mean +/- SE), as opposed to their respective number before therapy (6.9 +/- 0.5 and 8.3 +/- 0.8) (p less than 0.01). On the other hand, after treatment with cimetidine, the number of antral and fundic labeled cells was significantly higher (11.8 +/- 0.9 and 7.5 +/- 1.0, respectively) as compared with their number before therapy (5.7 +/- 0.5 and 5.6 +/- 0.6, respectively). The decreased gastric cell turnover induced by misoprostol indicates that the trophic effect of prostanoids on gastric mucosa is not due to an increase in cellular kinetics. The increased gastric cell turnover induced by cimetidine may contribute to its therapeutic effect in peptic ulcer disease.

  3. A study of the localized humoral immune response to implicated microorganisms in juvenile periodontitis

    Energy Technology Data Exchange (ETDEWEB)

    Hall, E.R.


    A study was undertaken using an in vitro explant culture system to determine the presence of immunoglobulins (IgG, IgA, and IgM) in the supernatant fluids (SF) of disease gingival tissue explant cultures. Studies were also undertaken to determine if the de novo biosynthesis of {sup 14}C-immunoglobulins could be observed in the explant cultures of diseased tissues from juvenile periodontitis (JP) patients. Radiolabeled proteins were detected in the SF and immunodiffusion studies using goat antihuman gamma, alpha or mu chain serum revealed the presence of IgG and IgA but no IgM present in the SF of the JP gingival tissue explant cultures. Immunodiffusion studies using goat anti-human gamma chain serum with Staph protein A isolated IgG fractions of the SF, followed by autoradiography of the IgG precipitation lines demonstrated the biosynthesis of IgG by the JP gingival tissue explant cultures. The serological studies suggested that local immune response in JP was to a polymicrobic infection. The SF of JP showed significantly higher levels of antibody reactivity to B. intermedius, C. ochracea, E. nodatum and P. micros as compared to healthy tissues. The local antibody response to the microorganisms tested differed from that observed in the sera of the patients.

  4. Specificity of indium-111 granulocyte scanning and fecal excretion measurement in inflammatory bowel disease--an autoradiographic study

    Energy Technology Data Exchange (ETDEWEB)

    Keshavarzian, A.; Price, Y.E.; Peters, A.M.; Lavender, J.P.; Wright, N.A.; Hodgson, H.J.


    The validity of /sup 111/In granulocyte scanning and fecal excretion measurement, as a reflection of loss of cells into the gastrointestinal tract, was studied using an autoradiographic technique in 11 patients in whom /sup 111/In granulocyte scan and colonoscopy were carried out simultaneously. /sup 111/In granulocytes were injected 1.5-4 hr prior to colonoscopy, and intraluminal fluid, mucosal brushings, and colonic biopsies were collected during the colonoscopy. In two patients with no histological evidence of inflammatory bowel disease, and four patients with clinically and histologically inactive inflammatory bowel disease, no /sup 111/Indium was detected in fluid, brushing, or biopsies. In five patients with active disease, 85% of the /sup 111/In activity in colonic fluid was precipitated by low-speed centrifugation. Autoradiography confirmed that the label remained attached to whole granulocytes in colonic fluid and mucosal brushings. Studies on biopsies, at intervals up to 4 1/2 hr following labeled granulocyte injection, demonstrated labeled polymorphonuclear neutrophils (PMNs) on the inflamed epithelial surface, with occasional cells in crypt abscesses by 110 min. We conclude that the techniques of /sup 111/In granulocyte scanning and fecal counting in patients with IBD are specifically measuring cell loss; labeled PMNs are capable of migrating through the gastrointestinal mucosa, in active disease, within 2 hr of administration.


    Institute of Scientific and Technical Information of China (English)

    余枭; 韩天权; 汤耀卿; 雷若庆; 夏宗勤


    Objective To evaluate the changes of pancreatic acinar cell functions in the rats with acute necrotizing pancreatitis (ANP). Methods Seventy SD rats were randomized into two groups: experimental group (n=35) and control group (n=35). To prepare the experimental model, the retrograde injection of 5% sodium taurocholate into the pancreatic duct was used for inducing ANP. Radioactive tracing by L- 3H-phenylalanine and autoradiography were performed for scoring the differences of changes of amino acid uptake, enzyme-protein synthesis and output from acinar cells in rats between both groups. Results No changes were observed in amino acid uptake and enzyme-protein synthesis in rats with dotted and haemorrhagic necrotizing foci as compared with control group. However, accumulated zymogen granules in the interstitial of acinar cells were seen in the experimental group. Conclusion It indicates that in experimental ANP rats, the functions of acinar cells in both amino acid uptake and protein synthesis were essentially normal, but the pathway of enzyme output was affected into ectopic secretion through the bottom or lateral cellular membrane of pancreatic acinar cell.

  6. Novel ¹⁸F-labeled benzoxazole derivatives as potential positron emission tomography probes for imaging of cerebral β-amyloid plaques in Alzheimer's disease. (United States)

    Cui, Mengchao; Ono, Masahiro; Kimura, Hiroyuki; Ueda, Masashi; Nakamoto, Yuji; Togashi, Kaori; Okamoto, Yoko; Ihara, Masafumi; Takahashi, Ryosuke; Liu, Boli; Saji, Hideo


    Two radiofluoro-pegylated phenylbenzoxazole derivatives, 4-(5-(2-(2-(2-[(18)F]fluoroethoxy)ethoxy)ethoxy)benzo[d]oxazol-2-yl)-N-methylaniline ([(18)F]24) and 4-(5-(2-(2-(2-[(18)F]fluoroethoxy)ethoxy)ethoxy)benzo[d]oxazol-2-yl)-N,N-dimethylaniline ([(18)F]32), were synthesized and evaluated as probes for imaging cerebral β-amyloid (Aβ) plaques in living brain tissue by PET. [(18)F]24 and [(18)F]32 displayed high affinity for Aβ(1-42) aggregates (K(i) = 9.3 and 3.9 nM, respectively). In vitro autoradiography with sections of post-mortem AD brain and transgenic mouse brain confirmed the affinity of these tracers. Initial high uptake into and rapid washout from the brain in normal mice were observed. [(18)F]24 also displayed excellent binding to Aβ plaques in ex vivo autoradiographic experiments with Tg2576 mice. Furthermore, small-animal PET studies demonstrated significant differences in the clearance profile after the administration of [(18)F]24 between Tg2576 and wild-type mice. The results suggest [(18)F]24 to be a useful PET agent for detecting Aβ plaques in the living human brain.

  7. Comparison of Cannabinoid CB1 Receptor Binding in Adolescent and Adult Rats: A Positron Emission Tomography Study Using [18F]MK-9470

    Directory of Open Access Journals (Sweden)

    Mathieu Verdurand


    Full Text Available Despite the important role of cannabinoid CB1 receptors (CB1R in brain development, little is known about their status during adolescence, a critical period for both the development of psychosis and for initiation to substance abuse. In the present study, we assessed the ontogeny of CB1R in adolescent and adult rats in vivo using positron emission tomography with [18F]MK-9470. Analysis of covariance (ANCOVA to control for body weight that would potentially influence [18F]MK-9470 values between the two groups revealed a main effect of age (F(1,109=5.0, P=0.02 on [18F]MK-9470 absolute binding (calculated as percentage of injected dose with adult estimated marginal means being higher compared to adolescents amongst 11 brain regions. This finding was confirmed using in vitro autoradiography with [3H]CP55,940 (F(10,99=140.1, P<0.0001. This ontogenetic pattern, suggesting increase of CB1R during the transition from adolescence to adulthood, is the opposite of most other neuroreceptor systems undergoing pruning during this period.

  8. Partial genetic deletion of neuregulin 1 and adolescent stress interact to alter NMDA receptor binding in the medial prefrontal cortex

    Directory of Open Access Journals (Sweden)

    Tariq Waseem Chohan


    Full Text Available Schizophrenia is thought to arise due to a complex interaction between genetic and environmental factors during early neurodevelopment. We have recently shown that partial genetic deletion of the schizophrenia susceptibility gene neuregulin 1 (Nrg1 and adolescent stress interact to disturb sensorimotor gating, neuroendocrine activity and dendritic morphology in mice. Both stress and Nrg1 may have converging effects upon N-methyl-D-aspartate receptors (NMDARs which are implicated in the pathogenesis of schizophrenia, sensorimotor gating and dendritic spine plasticity. Using an identical repeated restraint stress paradigm to our previous study, here we determined NMDAR binding across various brain regions in adolescent Nrg1 heterozygous (HET and wild-type (WT mice using [3H] MK-801 autoradiography. Repeated restraint stress increased NMDAR binding in the ventral part of the lateral septum (LSV and the dentate gyrus (DG of the hippocampus irrespective of genotype. Partial genetic deletion of Nrg1 interacted with adolescent stress to promote an altered pattern of NMDAR binding in the infralimbic (IL subregion of the medial prefrontal cortex. In the IL, whilst stress tended to increase NMDAR binding in WT mice, it decreased binding in Nrg1 HET mice. However in the DG, stress selectively increased the expression of NMDAR binding in Nrg1 HET mice but not WT mice. These results demonstrate a Nrg1-stress interaction during adolescence on NMDAR binding in the medial prefrontal cortex.

  9. Receptor Binding Sites for Substance P, but not Substance K or Neuromedin K, are Expressed in High Concentrations by Arterioles, Venules, and Lymph Nodules in Surgical Specimens Obtained from Patients with Ulcerative Colitis and Crohn Disease (United States)

    Mantyh, Christopher R.; Gates, Troy S.; Zimmerman, Robert P.; Welton, Mark L.; Passaro, Edward P.; Vigna, Steven R.; Maggio, John E.; Kruger, Lawrence; Mantyh, Patrick W.


    Several lines of evidence indicate that tachykinin neuropeptides [substance P (SP), substance K (SK), and neuromedin K (NK)] play a role in regulating the inflammatory and immune responses. To test this hypothesis in a human inflammatory disease, quantitative receptor autoradiography was used to examine possible abnormalities in tachykinin binding sites in surgical specimens from patients with inflammatory bowel disease. Surgical specimens of colon were obtained from patients with ulcerative colitis (n = 4) and Crohn disease (n = 4). Normal tissue was obtained from uninvolved areas of extensive resections for carcinoma (n = 6). In all cases, specimens were obtained germinal center of lymph nodules, whereas the concentrations of SP and SK binding sites expressed by the external muscle layers are not altered significantly. These results demonstrate that receptor binding sites for SP, but not SK or NK, are ectopically expressed in high concentrations (1000-2000 times normal) by cells involved in mediating inflammatory and immune responses. These data suggest that SP may be involved in the pathophysiology of inflammatory bowel disease and might provide some insight into the interaction between the nervous system and the regulation of inflammation and the immune response in human inflammatory disease.

  10. {sup 11}C-labeled stilbene derivatives as A{beta}-aggregate-specific PET imaging agents for Alzheimer's disease

    Energy Technology Data Exchange (ETDEWEB)

    Ono, Masahiro; Wilson, Alan; Nobrega, Jose; Westaway, David; Verhoeff, Paul; Zhuang Zhiping; Kung Meiping; Kung, Hank F. E-mail:


    A series of stilbene derivatives as potential diagnostic imaging agents targeting amyloid plaques in Alzheimer's disease (AD) were synthesized and evaluated. The syntheses of the stilbenes were successfully achieved by a simple Wadsworth-Emmons reaction between diethyl (4-nitrobenzyl)phosphonate and 4-methoxybenzaldehyde. 4-N,N-dimethylamino-4'-methyoxy and the corresponding 4-N-monomethylamino-, 4'-hydroxy stilbenes showed good binding affinities towards A{beta} aggregates in vitro (K{sub i} < 10 nM). The {sup 11}C labeled 4-N-methylamino-4'-hydroxystilbene, [{sup 11}C]4, was prepared by {sup 11}C methylation of 4-amino-4'-hydroxystilbene. The [{sup 11}C]4 displayed a moderate lipophilicity (log P = 2.36), and showed a very good brain penetration and washout from normal rat brain after an iv injection. In