WorldWideScience

Sample records for chloramine-t

  1. Preparation and evaluation of radio labelled recombinant human interleukin 2 by improved chloramine T method

    International Nuclear Information System (INIS)

    Iodination by improved chloramine T method was investigated. Iodine was activated with very small quantity of chloramine T (21 fold excess to iodine in molar ratio) and then recombinant interleukin-2 (rIL-2) was added to the mixture. Characteristics of the labelled rIL-2 after purification by reversed-phase HPLC was superior to those by usual chloramine T method or lactoperoxidase method. The specific activity was 0.74-1.11 MBq/μg and the immunoreactivity, biological activity and binding ability to rIL-2 receptors were similar to the intrinsic rIL-2. The improved chloramine T method is useful for iodination of unstable peptides and proteins. (author)

  2. Preparation and evaluation of radio labelled recombinant human interleukin 2 by improved chloramine T method

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Masahiro; Kominami, Goro; Kono, Masao (Shionogi and Co. Ltd., Osaka (Japan). Research Lab.)

    1991-03-01

    Iodination by improved chloramine T method was investigated. Iodine was activated with very small quantity of chloramine T (21 fold excess to iodine in molar ratio) and then recombinant interleukin-2 (rIL-2) was added to the mixture. Characteristics of the labelled rIL-2 after purification by reversed-phase HPLC was superior to those by usual chloramine T method or lactoperoxidase method. The specific activity was 0.74-1.11 MBq/{mu}g and the immunoreactivity, biological activity and binding ability to rIL-2 receptors were similar to the intrinsic rIL-2. The improved chloramine T method is useful for iodination of unstable peptides and proteins. (author).

  3. Pyridinium hydrobromide perbromide: a versatile catalyst for aziridination of olefins using Chloramine-T.

    Science.gov (United States)

    Ali, S I; Nikalje, M D; Sudalai, A

    1999-09-01

    [reaction: see text] Pyridinium hydrobromide perbromide (Py x HBr3) catalyzes effectively the aziridination of electron-deficient as well as electron-rich olefins using Chloramine-T (N-chloro-N-sodio-p-toluenesulfonamide) as a nitrogen source to afford the corresponding aziridines in moderate to good yields. PMID:16118868

  4. Survival of cool and warm freshwater fish following chloramine-T exposure

    Science.gov (United States)

    Gaikowski, M.P.; Larson, W.J.; Gingerich, W.H.

    2008-01-01

    Chloramine-T is presently available in the USA to control mortalities associated with bacterial gill disease or external columnaris only through an Investigational New Animal Drug Permit authorized by the US Food and Drug Administration (FDA). Its US approval hinges on FDA's acceptance of several key data, including those describing animal safety. Chloramine-T is presently applied in US aquaculture, by permit only, once daily on consecutive or alternate days for 1??h at 10 to 20??mg/L to control mortalities associated with bacterial gill disease or external columnaris. Our objective was to determine the safety of chloramine-T bath exposures at multiples of the proposed maximum treatment concentration (i.e., 0, 20, 60, 100, and 200??mg/L) administered on four consecutive days at 20????C to lake sturgeon Acipenser fulvescens, northern pike Esox lucius, and walleye Sander vitreum, or at 27????C to channel catfish Ictalurus punctatus, and largemouth bass Micropterus salmoides. All fish were tested as five to eight week old fry except for walleye and channel catfish which were tested as both fry and fingerling (fingerlings were at least four weeks older than the fry tested). Walleye and channel catfish were selected to evaluate the effects of life stage (fry vs. fingerling), temperature (walleye - 15, 20, or 25????C; channel catfish - 22, 27, or 32????C), exposure duration (60 vs. 180??min), and water chemistry (walleye only - reconstituted soft water vs. well water). Except for channel catfish fry, survival was significantly reduced only when fish were treated at 100 or 200??mg/L. Channel catfish fry survival was significantly reduced when exposed at 60??mg/L for 180??min at 27????C. Based on our mortality data, chloramine-T administered once daily for 60??min on four consecutive days at concentrations of up to 20??mg/L is not likely to adversely affect survival of cool or warmwater fish cultured in freshwater. Crown Copyright ?? 2007.

  5. Radioiodination of interleukin 2 to high specific activities by the vapor-phase chloramine T method

    International Nuclear Information System (INIS)

    Recombinant human interleukin 2 (IL-2) was radioiodinated utilizing the vapor phase chloramine T method of iodination. The method is rapid, reproducible, and allows the efficient radioiodination of IL-2 to specific activities higher than those previously attained with full retention of biological activity. IL-2 radioiodinated by this method binds with high affinity to receptors present on phytohemagglutinin-stimulated peripheral blood lymphocytes and should be useful for the study of receptor structure and function

  6. Acid-base and ionic fluxes in rainbow trout (Oncorhynchus mykiss) during exposure to chloramine-T

    Energy Technology Data Exchange (ETDEWEB)

    Powell, M.D.; Perry, S.F. [Department of Biology, University of Ottawa, 30 Marie Curie Ottawa, Ontario, K1N 6N5 (Canada)

    1998-09-01

    The effects of chloramine-T and its degradation products, sodium hypochlorite (NaOCl) and para-toluenesulphonamide (pTSA), on whole body acid-base and branchial and renal ion (Na{sup +}and Cl{sup -}) fluxes were examined in rainbow trout (Oncorhynchus mykiss). Exposure to chloramine-T (3.5 h, 18 mg l{sup -1}) resulted in increases in plasma total CO{sub 2} but no coincident rise in P{sub a}CO{sub 2} or reduction in blood pH. Exposure of fish to 2, 9 or 18 mg l{sup -1} chloramine-T (3.5 h duration) resulted in a reduction in net acid uptake suggesting the development of a metabolic alkalosis. Exposure to the chloramine-T breakdown product pTSA (dissolved in DMSO) resulted in increased net acid uptake (decreased acid excretion) suggesting a metabolic acidosis. Whole body ion fluxes demonstrated increases in the losses of both Na{sup +}and Cl{sup -} with chloramine-T, NaOCl and pTSA. However, the effect of DMSO alone could not be isolated. Confirmatory studies using fish in which the urinary bladder (to allow collection of urine) and dorsal aorta (to allow injection of [{sup 14}C]polyethylene glycol 4000 ([{sup 14}C]PEG), an extracellular fluid marker) were catheterised, revealed that changes in whole body ion fluxes during chloramine-T exposure could not be explained by increased renal efflux through urine flow, glomerular filtration or renal clearance. Branchial effluxes of [{sup 14}C]PEG were not significantly affected by chloramine-T exposure suggesting that the changes in whole body ionic fluxes were caused by transcellular rather than paracellular processes. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  7. Effect of chloramine-T labeling conditions on the stability of monoclonal antibodies and their fragments

    International Nuclear Information System (INIS)

    Rapid in vivo degradation of radioiodinated monoclonal antibodies (MAb) has been reported. Conditions for radioiodination have varied. The purposes of this study were to compare the stability of MAb and their fragments when iodinated with chloramine-T (CT) under different conditions, and to compare methods for quality assessment of the radioiodinated molecules. A B-cell lymphoma MAb (Lym-1, IgG2a) and its FAb fragment, and a mammary cancer MAb(B6.01, IgG1) and its F(Ab')/sub 2/ fragment were iodinated with I-125 at CT:AB and I:Ab ratios of 1:1 and 1:10. Molecular sieving (TSK-3000) high performance liquid chromatography (HPLC), cellulose acetate electrophoresis (CAE) at 11 and 45 minutes and solid phase immunoreactivity (IRA) were used to observe stability of the molecules when stored at 40C. Radiochemical yield was greater than 95% in all instances. Iodination at CT:Ab and I:Ab ratios of 1:1 induced progressive degradation in all species which was most marked for the fragments. Iodination at CT:Ab and I:Ab ratios of 1:10 resulted in no observable degradation over 21 days. There was no significant difference in degradation between the IgG2a and IgG1 antibody when iodinated under identical circumstances. HPLC, CAE for 11 minutes and IRA, but not CAE for 45 minutes, revealed comparable changes. The authors conclude that lesser amounts of chloramine-T can be used to iodinate MAb and their fragments without loss of radiochemical efficiency and with improved stability of the species. MAb fragments are more vulnerable to chloramine-T. These observations may explain, at least in part, rapid in vivo degradation of radioiodinated MAb

  8. Using micro-quantity of chloramine T in chicken pro-latin labelling and radioimmunoassay

    International Nuclear Information System (INIS)

    A radioimmunoassay was developed for measurement of chicken plasma prolactin. The assay used chPRL(AFP-10328B) as reference standard, chPRL(AFP-4444B) as the radio labelled ligand, rabbit anti-chicken PRL (AFP-151040789) as first antibody, and donkey anti-rabbit IgG antiserum as second antibody. For iodide ratio labelling of chicken PRL, a modified chloramine T method which reduced the amount of chloramine T and omitted sodium metabisulfite, and produced the labelled hormone with a specific activity of 29μCi/μg was used. The assay sensitivity was 0.34 ng/ml. the ED75, ED50 and ED25 of standard curve were 1.30, 3.71 and 10.60 ng/ml, respectively. Both coefficients of variations between and within assay were less than 15%. Serial dilutions of chicken samples showed a parallel inhibition curve to that of the standards. Plasma PRL concentrations of samples from hens at different reproductive states measured by this assay revealed significant differences and rational changing trends. These results validate the assay

  9. Inhalation exposure to chloramine T induces DNA damage and inflammation in lung of Sprague-Dawley rats.

    Science.gov (United States)

    Shim, Ilseob; Seo, Gyun-Baek; Oh, Eunha; Lee, Mimi; Kwon, Jung-Taek; Sul, Donggeun; Lee, Byung-Woo; Yoon, Byung-Il; Kim, Pilje; Choi, Kyunghee; Kim, Hyun-Mi

    2013-01-01

    Chloramine T has been widely used as a disinfectant in many areas such as kitchens, laboratories and hospitals. It has been also used as a biocide in air fresheners and deodorants which are consumer products; however, little is known about its toxic effects by inhalation route. This study was performed to identify the subacute inhalation toxicity of chloramine T under whole-body inhalation exposure conditions. Male and female groups of rats were exposed to chloramine T at concentrations of 0.2, 0.9 and 4.0 mg/m³ for 6 hr/day, 5 days/week during 4 weeks. After 28-day repeated inhalation of chloramine T, there were dose-dependently significant DNA damage in the rat tissues evaluated and inflammation was histopathologically noted around the terminal airways of the lung in both genders. As a result of the expression of three types of antioxidant enzymes (SOD-2, GPx-1, PRX-1) in rat's lung after exposure, there was no significant change of all antioxidant enzymes in the male and female rats. The results showed that no observed adverse effect level (NOAEL) was 0.2 mg/m³ in male rats and 0.9 mg/m³ in female rats under the present experimental condition.

  10. Spectrophotometric determination of bio-active compounds with chloramine-T and gallocyanine.

    Science.gov (United States)

    Sastry, C S; Srinivas, K R; Prasad, K M

    1996-10-01

    A simple, sensitive and selective method for the spectrophotometric determination of drugs, viz., sulphamethoxazole, tetracycline HCl, amidopyrine, nifurtimox and isoniazid and biologically important amino acids, cysteine, aspartic acid and arginine based on their reactivity with chloramine-T (CAT) is proposed. The method involves the addition of excess CAT of a known concentration in the presence of 0.25 M HCl and the determination of the unreacted CAT by measurement of the decrease in the absorbance of the dye, gallocyanine (lambda(max): 540 nm), the most suitable of several dyes that were tested. This method was applied to the determination of drug contents in pharmaceutical formulations and to the measurement of the aspartic acid content of some protein hydrolysates. The method is useful for the determination of the target compounds in microgram quantities from 0.4-5.6 microg mL(-1) with the exceptions of arginine (1.0-8.0 microg mL(-1)) and nifurtimox (0.8-5.6 microg mL(-1)). Standard deviations were typically 0.5 mg per dose (RSD 0.5-1.2%). No interferences were observed from common excipients in formulations, and detailed interference studies of other amino acids in the determination of cysteine, aspartic acid and arginine are reported. The validity of the method was tested against spectrophotometric and titrimetric reference methods. Recoveries were 99.8-102.1%. PMID:18966644

  11. Mechanistic chemistry of oxidation of balsalazide with acidic chloramine-T and bromamine-T: A comparative spectrophotometric kinetic study

    Indian Academy of Sciences (India)

    Puttaswamy; S Dakshayani

    2014-11-01

    Balsalazide (BSZ) belongs to a class of non-steroidal anti-inflammatory drugs. Kinetics and mechanism of oxidation of BSZ with sodium N-halo-p-toluenesulfonamides viz., chloramine-T(CAT) and bromamine-T(BAT) in HClO4 medium have been spectrophotometrically investigated (max =357nm) at 303 K. Under comparable experimental conditions, reactions with both the oxidants follow a first-order dependence of rate on [BSZ] and fractional-order dependence on each [oxidant] and [HClO4]. Activation parameters and reaction constants have been computed. 2-hydroxy-5-nitroso-benzoic acid and 3-(4-nitroso-benzoylamino)-propionic acid are identified as the oxidation products of BSZ with both CAT and BAT. The rate of oxidation of BSZ is about five-fold faster with BAT than with CAT. Plausible mechanism and related rate law have been deduced for the observed kinetics.

  12. Histopathology of repeated, intermittent exposure of chloramine-T to walleye (Sander vitreum) and (Ictalurus punctalus) channel catfish

    Science.gov (United States)

    Gaikowski, M.P.; Densmore, Christine L.; Blazer, V.S.

    2009-01-01

    Chloramine-T (Cl-T) has been used safely and effectively to control bacterial gill disease in salmonids at a maximum exposure regimen of up to four consecutive, once-daily exposures administered for 60??min at 20??mg/L. However, data to document safe treatment concentrations of Cl-T are lacking for freshwater-reared fish other than salmonids. We report the histopathology resultant from the administration of 12 consecutive, once-daily, 180-min static immersion baths of 0, 20, 50, or 80??mg Cl-T/L to walleye (20????C) and channel catfish (27????C). Twelve fish of each species were euthanized immediately before the first exposure (initial controls) and then after the twelfth exposure and 7 and 14??days after the twelfth exposure. Only initial controls and fish euthanized immediately after the twelfth exposure were processed for histological review because of the general lack of exposure-related lesions in exposed fish. The only exposure-related histological changes were in the spleen where significantly greater erythrocyte swelling and necrosis was observed in channel catfish exposed at 80??mg/L relative to exposure at 0??mg/L; similar histological changes were insignificant for walleye, though there appeared to be a shift in the general category of histological change with degenerative changes (necrosis, etc.) observed following exposure at 50 or 80??mg/L compared to the inflammatory and hemodynamic changes (congestion, leukocyte infiltrate, etc.) observed in walleye exposed at 0 or 20??mg/L. The only significant change in peripheral blood cytology was that walleye fingerlings exposed at 80??mg/L had significantly fewer mature red blood cells and significantly more immature red blood cells per oil-immersion field than controls. The histopathological changes observed following exposure to Cl-T under an exaggerated exposure regimen suggest that walleye or channel catfish therapeutically exposed to Cl-T will not have treatment-related histological changes.

  13. Influence of chloramine T iodination on the biological and immunological activity or the molecular radius of the human growth hormone molecule

    International Nuclear Information System (INIS)

    Potential alterations of the somatotropic activity of human growth hormone (hGH) resulting from Chloramine T labelling reaction, iodination up to 2.7 atoms/molecule and indirect radiation effects, have been studied. Three 2X2 factorial assays, performed in hypophysectomized rats, failed to reveal any significant difference (P greater than 0.05) in true growth promoting activity between hGH and (127-I)hGH, even after storing the latter with 125-I. Similar results were obtained applying a sensitive and precise gel filtration technique for Stokes Radius determination and radioimmunoassay

  14. Effects of sub lethal concentration of Chloramin T on growth, survival, haematocrit and some blood biochemical parameters in common carp fry (Cyprinus carpio

    Directory of Open Access Journals (Sweden)

    Mohamad R. Imanpoor

    2011-07-01

    Full Text Available This study was done in Gorgan University of Agricultural Sciences and Natural Resources, in 2009, during 8 weeks to survey effects of different concentrations of Chloramin T on fry common carp (Cyprinus carpio (Linnaeus, 1758. According to the pre experiment data, lethal concentration, the lowest observed effect concentration, maximum allowable toxicant concentration and No Observed Effect Concentration of Chloramin T (Halamid in common carp (C. carpio fry were respectively 40.9, 27.1, 4.90 and 11.28 mg/L-1. Hence, the range of our experiment was between 0 to 25 mg L-1 which was divided to five treatments (0, 5, 10, 15 and 25 mg/L-1. At the end of experiment we calculated growth factor (special growth rate and food conversion ratio and stress indices (glucose total protein and percent of haematocrit and then they were compared with the controlled group. Our study results showed no significant difference between percentage of increase in body weight (280.4±25.79 - 200.4±10.16, special growth rate (2.34±0.25 - 1.96±0.06 and food conversion ratio (0.84±0.05 - 0.60±0.12 in experimental lots of fishes. There is also no significant difference between glucose (146.82±0.79 - 99.54±1.89 and total protein (3.61±0.45 - 2.82±0.06 in experimental groups (P>.05. However there is a significant difference between the percentages of haematocrit (57.34±4.99 - 40.74±2.17 and cholesterol (362.05±24.38 - 134.92±17.59 in these groups (P<0.05.

  15. Fluorous Analogue of Chloramine-T: Preparation, X-ray Structure Determination, and Use as an Oxidant for Radioiodination and s-Tetrazine Synthesis.

    Science.gov (United States)

    Dzandzi, James P K; Beckford Vera, Denis R; Genady, Afaf R; Albu, Silvia A; Eltringham-Smith, Louise J; Capretta, Alfredo; Sheffield, William P; Valliant, John F

    2015-07-17

    A fluorous oxidant that can be used to introduce radioiodine into small molecules and proteins and generate iodinated tetrazines for bioorthogonal chemistry has been developed. The oxidant was prepared in 87% overall yield by combining a fluorous amine with tosyl chloride, followed by chlorination using aqueous sodium hypochlorite. A crystal structure of the oxidant, which is a fluorous analogue of chloramine-T, was obtained. The compound was shown to be stable for 7 days in EtOH and for longer than three months as a solid. The oxidant was effective at promoting the labeling of arylstannanes using [(125)I]NaI, where products were isolated in high specific activity in yields ranging from 46% to 86%. Similarly, iodinated biologically active proteins (e.g., thrombin) were successfully produced, as well as a radioiodinated tetrazine, through a concomitant oxidation-halodemetalation reaction. Because of its fluorous nature, unreacted oxidant and associated reaction byproducts can be removed quantitatively from reaction mixtures by passing solutions through fluorous solid phase extraction cartridges. This feature enables rapid and facile purification, which is critical when working with radionuclides and is similarly beneficial for general synthetic applications. PMID:26030355

  16. Use of chloramine-T and two dyes in the sensitive determination of stavudine in pharmaceuticals Uso de cloramina-T e de dois corantes na determinação sensível de estavudina em medicamentos

    Directory of Open Access Journals (Sweden)

    Kanakapura Basavaiah

    2007-09-01

    Full Text Available Three new methods are described for the assay of stavudine (STV in bulk drug and in dosage forms using chloramine-T (CAT and two dyes, methyl orange and indigocarmine, as reagents. Titrimetry involves treating STV with a measured excess of CAT in hydrochloric acid medium, and after the oxidation of STV is judged to be complete, the unreacted oxidant is determined iodometrically. Spectrophotometric methods entail the addition of a known excess of CAT to STV in hydrochloric acid medium followed by determination of residual oxidant by reacting with a fixed amount of either methyl orange and measuring the absorbance at 520 nm (Method A or indigo carmine and measuring the absorbance at 610 nm (Method B. In all the methods, the amount of CAT reacted corresponds to the amount of STV. In titrimetric method, the reaction follows 1:1 stoichiometry (STV: CAT, and is applicable over the range 1.5-10 mg of STV. In spectrophotometric methods, the absorbance is found to increase linearly with concentration of STV. The systems obey Beer's law for 0.2-2.0 and 1.0-10.0 mg/mL for method A and method B, respectively. The apparent molar absorptivities are calculated to be 5.7x10(4 and 1.5x10(4 L/mol/cm for method A and method B, respectively, and the corresponding Sandell sensitivity values are 0.004 and 0.015 µg/cm². The limits of detection and quantification are reported for both methods. Intra-day and inter-day precision and accuracy of the developed methods were evaluated as per the current ICH guidelines. The methods were successfully applied to the assay of STV in tablet and capsule formulations and the results were compared with those of a reference method by applying Student's t-test and F-test. No interference was observed from common tablet adjuvants. The accuracy and reliability of the methods were further ascertained by performing recovery experiments via standard-addition method.Descrevem-se três novos métodos para o ensaio de estavudina (STV na mat

  17. Preparation of iodine - 125 - labeled insulin for radioimmunoassay: comparison of chloramine T and iodogen iodination

    International Nuclear Information System (INIS)

    Stoichiometric iodination of porcine insulin was performed to the general method of Hunter and Greenwood with modifications recommended by Roth. These method was compared with radioidination using Iodogen. Films of Iodogen react rapidly in the solid phase with aqueous mixtures of I -and proteins. For two methods satisfactory activity of the labeled porcine insulin was obtained and characteristics of the radioimmunoassay were studied. (author)

  18. 氯胺T氧化孔雀绿动力学光度法测定食品中痕量碘%Kinetic-Spectrophotometric Determination of Trace Iodine in Food with the Oxidation Reaction of Malachite Green by Chloramine T

    Institute of Scientific and Technical Information of China (English)

    李建国; 乔艳; 魏永前

    1999-01-01

    基于在稀盐酸介质中,碘催化氯胺T氧化孔雀绿而使其褪色的反应,建立了测定痕量碘的新方法.测定碘的线性范围为0~80 μg/L;检出限为1.6μg/L.方法灵敏、简便、选择性好,用于食品中痕量碘的测定,结果满意.

  19. Chloramine-induced anaphylaxis while showering: a case report

    Directory of Open Access Journals (Sweden)

    D’Alò Simona

    2012-09-01

    Full Text Available Abstract Introduction Sodium-N-chlorine-p-toluene sulfonamide, commonly known as chloramine-T, is a derivative of chlorine which is widely used as a disinfectant. For many years, chloramine-T has been described as a cause of immediate-type hypersensitivity, especially with regard to asthma and rhinitis, and as a cause of occupational dermatoses in cleaning personnel in hospitals, although no anaphylactic reaction has yet been reported. Hence, to the best of our knowledge we present the first case of anaphylaxis to chloramine-T with evidence of specific immunoglobulin E antibodies. Case presentation We describe the case of a 25-year-old Caucasian woman who was in good health and with a negative history for atopy, including no respiratory symptoms of rhinitis or asthma, and with no professional exposure to chloramine-T. She, while showering, applied a chloramine-T solution to a skin area with folliculitis on her leg, and within a few minutes developed generalized urticaria and angioedema, followed by vomiting and collapse with loss of consciousness. A skin prick test with a chloramine-T solution at 10mg/mL concentration was positive, and specific immunoglobulin E to chloramine-T was quantified at a value of 2.9 optical density as measured by the enzyme allergosorbent test technique. Conclusion The strict cause-effect relationship and the results of the skin test and the in vitro test make certain the causative role of chloramine-T in this case of anaphylaxis. This suggests that chloramine-T, based on its wide use as a disinfectant, should be considered a possible cause in anaphylaxis of unknown origin.

  20. Radio-labeling of polyethylene glycol modification of recombinant human interleukin-6

    International Nuclear Information System (INIS)

    Iodine-125 was used as labeling nuclide, and the PEG-rhIL-6 was labeled by the common used chloramines-T and the two-phase chloramines-T, respectively. The labeled compound was purified by both methods of gel filtration and ultrafiltration respectively. The purity of the labeled PEG-rhIL-6 was determined by both trichloroacetic acid (TCA) and SDS-PAGE, and the biological activity was determined by MTT method. The results demonstrated that the labeling rate and specific radioactivity were 74.5% and 5.513 x 105 Bq/μg for PEG-rhIL-6 by the two-phase chloramines-T method, higher than that by the common used chloramines-T method, which was 62.3% and 4.610 x 105 Bq/μg respectively. The purity of labeled PEG-rhIL-6, purified by both gel filtration and ultrafiltration methods, was over 99% with TCA method. The labeled PEG-rhIL-6 by two-phase chloramines-T method showed two bands, which was identical to that of standard PEG-rhIL-6 though SDS-PAGE, but the labeled PEG-rhIL-6 by common used chloramines-T method had one more band compared with standard PEG-rhIL-6. When determined by MTT, it shown that the biological activity of PEG-rhIL-6 iodinated by common used chloramines-T method was lower than that by two-phase chloramines-T method. (authors)

  1. Preparation and characterization of 125 I labeled bovine serum albumin

    Directory of Open Access Journals (Sweden)

    K S Ashwitha Rai

    2015-01-01

    Full Text Available Bovine serum albumin is a model protein, which has been conventionally used as protein standard and in many areas of biochemistry, pharmacology and medicine. Radioiodination procedure for bovine serum albumin employing chloramine-T as an oxidant with slight modification was evaluated critically to establish the optimal conditions for the preparation of radiolabeled tracer ( 125 I-BSA with required specific activity without impairing the immune reactivity and biological activity. Optimized radioiodination procedure involving 10 µg of chloramine-T along with 20 µg of sodium metabisulphite with 60 seconds incubation at 2° yielded 125 I-BSA with high integrity.

  2. One-pot synthesis of new series 3,4,5-trisubstituted-dihydroisoxazoline derivatives via 1,3-dipolar cycloaddition of nitrile oxides with chalcones

    Indian Academy of Sciences (India)

    Raad Kasim Yhya; K M Lokanatha Rai; Ebraheem Abdu Musad

    2013-07-01

    We have synthesized a series of novel isoxazolines via 1,3-dipolar cycloaddition reaction. Aromatic aldoximes undergo oxidative-dehydrogenation with chloramine-T to give nitrile oxides, which were reacted with chalcones to afford of 3,4,5-trisubstituted 4,5-dihydroisoxazolines in a good yield.

  3. Improved radioimmunotherapy of hematologic malignancies. Progress report, November 1, 1993--October 31, 1994

    Energy Technology Data Exchange (ETDEWEB)

    Press, O.W.

    1994-08-04

    This report summaries progress made during the time interval between November 1, 1993 and October 31, 1994 and briefly describes studies on the metabolism of antibodies targeting B cell antigens, retention of labeled antibodies by human B cell lymphocytes, and tissue distribution of Chloramine T and tyramine cellobiose labeled antibodies in mice harboring a human erythroleukemia tumor transplant.

  4. Synthesis of new series of 4, 5-dihydroisoxazole-5-carboxylate derivatives for the study of their liquid crystalline properties

    Indian Academy of Sciences (India)

    SUMANA Y KOTIAN; NARAYANA U KUDVA N; K M LOKANATHA RAI; K BYRAPPA

    2016-07-01

    A new series of 4,5-dihydroisoxazole-5-carboxylate derivatives were synthesized via [3+2] cycloaddition reaction between ethyl acrylate and nitrile oxide generated in situ in presence of Chloramine-T. The synthesized derivatives were characterized by Mass, IR and NMR Spectroscopy and their mesomorphic behavior were studied using DSC and Polarising Optical Microscopy.

  5. Synthesis and radioiodination of analogues of substance P for the building up of a radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Bienert, M.; Schmidt, H.E.; Ehrlich, A.; Forner, K.; Klauschenz, E.; Furkert, J.; Rathsack, R.; Niedrich, H. (Akademie der Wissenschaften der DDR, Berlin. Inst. fuer Wirkstofforschung; Akademie der Wissenschaften der DDR, Berlin-Buch. Zentralinstitut fuer Isotopen- und Strahlenforschung)

    1984-02-01

    The substance P (SP) analogs (Lys(MSOC)/sup 3/)-SP, (Tyr/sup 8/)-SP, (Tyr/sup 8/,Nle/sup 11/)-SP and (p-HPA/sup 1/)-SP have been synthesized by classical methods of peptide synthesis as well as by the liquid phase peptide synthesis (LPPS) to allow conjugation with protein in the N..cap alpha..-position and radioiodination. (Tyr/sup 8/)-SP and (p-HPA-Arg/sup 1/)-SP have been radioiodinated by the chloramine T- and the lactoperoxidase method. A complete S-oxidation of SP was observed, when the chloramine T procedure was used, but this modification does not disturb the assay. The introduction of the (/sup 125/I-pHPA-Arg/sup 1/)-SP-tracer led to considerable improvements of our SP-radioimmunoassay.

  6. Labelling of human serum albumin with iodine-131 for diagnosis in nuclear medicine

    International Nuclear Information System (INIS)

    Labelling of 131I-human serum albumin with I-131 from a solution of 131I-sodium iodide using chloramine T as an oxidant agent is studied. Parameters which can influence on the labelling yield like mass of human serum albumin, and chloramine T, pH of the reaction, reaction time and activity of 131I are also studied. The purification of the labeled product by means of IRA-410 Amberlite ion-exchange resin in chloride form and the sterilization of the 131I-human serum albumin by its passage through a 0,22μ millipore filter are carried out. The radiochemistry control of the final product by paper chromatography and the microbiological control by cultivation of microorganisms in fluid medium: nutrient broth, sodium thioglycollate broth and Sabouraud, are performed. The stability of the radiopharmaceutical until ten days after its preparation is analysed by means of radiochemical control. (Author)

  7. Kinetics and Mechanism of Electron Transfer Reaction: Oxidation of Sulfanilic Acid by N-Chloro-p-Toluene Sulfonamide in Acid Perchlorate Medium

    Energy Technology Data Exchange (ETDEWEB)

    Sailani, Riya; Bhasin, Meneka; Khandelwal, C. L.; Sharma, P. D. [Univ. of Rajasthan, Jaipur (India)

    2014-01-15

    The kinetics and mechanism of oxidation of sulfanilic acid by N-chloro-p-toluene sulfonamide (chloramine-T) have been studied in acid medium. The species of chloramine-T were analysed on the basis of experimental observations and predominantly reactive species was taken into account for proposition of most plausible reaction mechanism. The derived rate law (1) conforms to such a mechanism. All kinetic parameters were evaluated. Activation parameters such as energy and entropy of activation were calculated to be (61.67 ± 0.47) kJ mol{sup -1} and (-62.71 ± 2.48) JK{sup -1}mol{sup -1} respectively employing Eyring equation.

  8. The effect of chemical anti-inhibitors on fibrinolytic enzymes and inhibitors

    DEFF Research Database (Denmark)

    Sidelmann, Johannes Jakobsen; Jespersen, J; Kluft, C;

    1997-01-01

    proteases. We studied the influence of chemical anti-inhibitors (chloramine T, flufenamate, sodium lauryl sulfate, and methylamine) on fibrinolytic serine proteases and fibrinolytic enzyme inhibitors using the physiological substrate fibrin as plasmin substrate. Low concentrations of chloramine T (0.01 mmol....../l) prevent the inhibition of plasminogen activators. Higher concentrations (1 mmol/l) reduce the inhibition of plasmin, but simultaneously quench the plasminogen activator activity. Flufenamate eliminates most fibrinolytic enzyme inhibitors, but increases the activity of plasmin (apparent recovery 140......Fibrinolytic enzyme inhibitors hamper the determination of the specific fibrinolytic serine protease activity. Reportedly, chemical anti-inhibitors eliminate the influence of fibrinolytic inhibitors, but it remains unclear to what extent they change the specific activity of fibrinolytic serine...

  9. Preparation and Evaluation of (125I) Daunorubicin as a Potential Agent for Tumor Detection and radiotherapy

    International Nuclear Information System (INIS)

    In this study, the optimization of daunorubicin labeling with iodine-125 and its biological evaluation were described. Daunorubicin was labeled via direct electrophilic substitution using chloramine-T as oxidizing agent. The optimum amounts of reactants were: 40μg daunorubicin, 30μg Chloramine-T and ∼ 19 KBq carrier free Na125I. The labeled daunorubicin was stable for more than 24 hours. Results of the in-vivo evaluation revealed that the tracer, [125I] daunorubicin, tends to localize in tissues with high proliferation rate with preferential accumulation in cancerous tissues. Imaging should be carried at 3 hours post injection. The in-vitro cell growth inhibition assay showed that the effect of [125I] Daunorubicin was stronger than the effect of cold daunorubicin which strongly suggested that its cytotoxicity was mainly due to radiotoxicity rather than chemotherapeutic activity.

  10. Human growth hormone (HGH), ch. 6

    International Nuclear Information System (INIS)

    A radioimmunoassay method for the human growth hormone (HGH) is described. The requirements are discussed in detail and a scheme for the preparation of incubation mixtures is given. HGH is labelled with 125I by the chloramine T method and purified by gel filtration or electrophoresis. Separation of bound and free-labelled hormones is performed by absorption of the free hormone, using talc or charcoal

  11. New aquaculture drugs under FDA review

    Science.gov (United States)

    Bowker, James D.; Gaikowski, Mark P.

    2012-01-01

    Only eight active pharmaceutical ingredients available in 18 drug products have been approved by the U.S. Food and Drug Administration for use in aquaculture. The approval process can be lengthy and expensive, but several new drugs and label claims are under review. Progress has been made on approvals for Halamid (chloramine-T), Aquaflor (florfenicol) and 35% PeroxAid (hydrogen peroxide) as therapeutic drugs. Data are also being generated for AQUI-S 20E, a fish sedative.

  12. Ethyl 4,4''-Difluoro-5'-hydroxy-1,1':3',1''-terphenyl-4'-carboxylate

    Directory of Open Access Journals (Sweden)

    Badiadka Narayana

    2011-11-01

    Full Text Available A simple and novel route for the synthesis of new terphenyl derivative as well as oxidative aromatization of α,β-unsaturated cyclohexenone to the corresponding phenol derivative is developed. The present work involves the condensation of ethylacetoacetate with 4,4'-difluoro chalcone followed by the aromatization using chloramine-T in acetic acid to yield the title compound (3. The synthesized compound (3 is well characterized by IR, NMR, LCMS and elemental analysis.

  13. Ethyl 4,4''-Difluoro-5'-hydroxy-1,1':3',1''-terphenyl-4'-carboxylate

    OpenAIRE

    Badiadka Narayana; Balladka Kunhanna Sarojini; Seranthimata Samshuddin

    2011-01-01

    A simple and novel route for the synthesis of new terphenyl derivative as well as oxidative aromatization of α,β-unsaturated cyclohexenone to the corresponding phenol derivative is developed. The present work involves the condensation of ethylacetoacetate with 4,4'-difluoro chalcone followed by the aromatization using chloramine-T in acetic acid to yield the title compound (3). The synthesized compound (3) is well characterized by IR, NMR, LCMS and elemental analysis.

  14. Comparison and evaluation of different methods for α-MSH labelling

    International Nuclear Information System (INIS)

    The authors have studied the behaviour of 125I-labelled α-MSH under different experimental conditions. Until now, the chloramine T method had been used by most investigators with variable results. They have tested three other labelling techniques based on 125I mild oxidation: (1) an enzymatic method with lactoperoxidase, (2) a sparingly soluble chloramine method (T.D.G.U.), and (3) modified chloramine T procedure, 'the iodine volatilization method'. Labelled hormone obtained after each kind of iodination was assayed for immunoreactivity. In addition, time course degradation was measured by classical RIA incubation procedures. Charcoal-dextran was used to separate bound and free antigen. The authors have found chloramine T-iodinated α-MSH to be significantly more damaged than preparations obtained by other methods and to be less stable when stored at -180C. No differences were found between the differently labelled 125I-labelled α-MSH fresh preparations in binding to surface receptors of human melanoma cell lines in culture. (Auth.)

  15. Preparation of iodine-123 labeled AM251: a potential SPECT radioligand for the brain cannabinoid CB1 receptor

    Energy Technology Data Exchange (ETDEWEB)

    Lan, Ruoxi; Makriyannis, Alexandros [Connecticut Univ., Molecular and Cell Biology Dept., Storrs, CT (United States); Gatley, S.J. [Brookhaven National Lab., Medical Dept., Upton, NY (United States)

    1996-10-01

    We report the synthesis and labeling with iodine-123 of N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251). This compound is an analog of the recently described cannabinoid receptor antagonist, SR141716A, in which a 4-chlorophenyl group is replaced by 4-iodophenyl. Labeling in good yield (62%) and radiochemical purity (> 95%), and high specific activity (> 2500 Ci/mmol) was achieved by an iododestannylation reaction using the tributyltin precursor, no carrier added I-123 iodide, and chloramine-T. (author).

  16. Chemical Modification of Amino Acid Residues in Human Plasminogen

    Institute of Scientific and Technical Information of China (English)

    LIANG Fang; SUN Hong; ZHAO Cheng-guang; CUI Ting; HONG Shui-sheng; CHEN Jia; LIU Lan-ying

    2003-01-01

    The chemical modification of human plasminogen(HPg) was studied with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide(EDC), N-acetylimidazole(NAI), 1,2-cyclohexanedione(CHD), chloramine T(Ch-T) and N-bromosuccinimide(NBS) as modifying reagents at its carboxyl group, tyrosine, arginine, methionine and tryptophan residues, respectively. The results indicate that tyrosine and arginine residues are not essential for HPg activity, while carboxyl groups, methionine and tryptophan residues are important for the activity of HPg. The Keech and Farrant′s kinetic analysis reveals that one tryptophan residue, one methionine residue and two carboxyl groups are essential for HPg activity.

  17. The tritium kinetic isotope effect for the iodination of L-(5-3H) -3-iodotyrosine

    International Nuclear Information System (INIS)

    The kinetic tritium isotope effect k(subH)/k(subT) for the iodination of L-(5-3H)-3-iodotyrosine at pH 7.5 was found to be 4.29 +-0.25 with molecular iodine and 4.50 +- 0.26 with chloramine-T/potassium iodide. This effect establishes that the rate-limiting step is proton removal by the general base. A novel method for measuring the fraction of tritiated water by high performance liquid chromatography is described

  18. Studies on the Radioimmunoassay of Human Growth Hormone - 1. Evaluation of the method of determination

    International Nuclear Information System (INIS)

    Utilizing the commercial radioimmunoassay kit, the author assayed HGH and evaluated the problems of the method. The method had the sensitivity of 0.5 mug/ml degree and could determine the plasma HGH concentration directly without the help of plasma extraction. Also it was specific for the HGH, when tested by a dilution method using test utilizing the plasma of the acromegalic patient. The author also obtained the 125I labelled HGH of specific activity of 156. 3 μCi/μg, performing the chloramine-T method.

  19. Isolation of a somatomedin binding protein from human preterm amniotic fluid: development of a radioimmunoassay

    International Nuclear Information System (INIS)

    This thesis investigates the nature and biological behaviour of a somatomedin binding protein, identified in preterm amniotic fluid (AF). For that purpose a double antibody radioimmunoassay was developed. Purified AF binding protein (AFBP) was iodinated by the chloramine-T method, and dilutions of partially purified AFBP were designated as the standard, with the results expressed in μg equivalent protein/ml. The sensitivity of the assay was improved by adoption of the nonequilibrium procedure. AFBP values were twice as high in preterm AF as in term AF. (Auth.)

  20. Qualitative and quantitative evaluation of donkeys responses to immunization by rabbits' IgG

    International Nuclear Information System (INIS)

    In this study two apparently healthy donkeys were immunized with highly pure rabbit's 1gG using a revised protocol. Qualitative test using the same immuno gen was done as a primary test to eva lute the immune system response. However, the same 1gG was iodinated with 125I using chloramine T method and the labeled 1gG was used to quantitatively study the immune response. The two donkeys showed good response with the younger one having the best response. The obtained donkey anti rabbit sera was used as separating agent for RIA assay for human PRL. (Author)

  1. optimization of insulin radioreceptor assay in human erythrocytes in normal and some disease status

    International Nuclear Information System (INIS)

    This study is concerned with the evaluation of a new optimized technique for the principle of chloramine-T method used for insulin iodination by 125I-radioisotope with some modifications. The modified procedure can be carried out under normal condition of room temperature, employed longer reaction times and omitted the addition of inorganic reducing salts, maintaining efficient iodination and avoiding denaturations to obtain labels of exceedingly high specific activity on a small quantities of insulin for in vitro usage in the investigation of human erythrocytes 125 I-insulin binding capacity in normal and some disease status

  2. Preparation of iodine-123 labeled AM251: a potential SPECT radioligand for the brain cannabinoid CB1 receptor

    International Nuclear Information System (INIS)

    We report the synthesis and labeling with iodine-123 of N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251). This compound is an analog of the recently described cannabinoid receptor antagonist, SR141716A, in which a 4-chlorophenyl group is replaced by 4-iodophenyl. Labeling in good yield (62%) and radiochemical purity (> 95%), and high specific activity (> 2500 Ci/mmol) was achieved by an iododestannylation reaction using the tributyltin precursor, no carrier added I-123 iodide, and chloramine-T. (author)

  3. Comparison of 131I-TYR3-octreotate and 131I-DOTA-TYR3-octreotate: The effect of DOTA on pharmacokinetics and stability

    International Nuclear Information System (INIS)

    The authors compared the biodistribution, and in vivo and in vitro stabilities of 131I-Tyr3-octreotate and 131I-DOTA-Tyr3-octreotate. The peptides were radioiodinated by the chloramine T method and high radiochemical yields were obtained (greater than 97%). Both labelled compounds showed high stability when incubated in human plasma at 37 deg C. The 131I-Tyr3-octreotate showed significant hepatic uptake and biliary excretion. The biodistribution of 131I-DOTA-Tyr3-octreotate, however, can be compared with the distribution of radiometal labelled octreotide analogues. (author)

  4. Comparison of specific radioactivities of human alpha-lactalbumin iodinated by three different methods

    Energy Technology Data Exchange (ETDEWEB)

    Thean, E.T. ( Monash Medical School, Prahran, Victoria (Australia))

    1990-08-01

    Radioiodination provides an extremely sensitive method for the detection of low levels of proteins. In the development of a sensitive radioimmunoassay for human alpha-lactalbumin (alpha-LA), the protein was labeled to high specific activity (approaching 2000 Ci/mmol) with lactoperoxidase, chloramine-T, and Iodogen. Despite high specific activities of the labeled protein by each method, there was a considerable difference in their binding affinity with monoclonal anti-human alpha-LA antibodies due to varying degrees of protein damage. Iodination of human alpha-LA with Iodogen resulted in labels of the highest specific activity and immunoreactivity with the monoclonal antibodies used.

  5. The experience with setting-up radioimmunoassay for alpha-1 fetoprotein

    International Nuclear Information System (INIS)

    The decisive factor in the preparation of radioimmunological alpha-1-fetoprotein determination, provided sufficient commercial or own antisera and standards are available for calibration, is the quality of the preparation for labelling. Alpha-1-fetoprotein was separated by affinity chromatography using Sepharose with alpha-1-fetoprotein-bound antibodies. The isolates thus obtained were labelled with 125I using enzyme and chloramine T and Iodogen techniques. The labelled alpha-1-fetoprotein can be used for RIA. In view of reduced immunoreactivity of the preparation, however, the performance of the radioimmunological determination has so far not matched the quality of imported kits. The technique is currently being optimized. (author)

  6. Radioiodination and biological evaluation of levalbuterol as a new selective radiotracer. A β{sub 2}-adrenoceptor agonist

    Energy Technology Data Exchange (ETDEWEB)

    Sanad, Mahmoud Hamdi; Abelrahman, Mohamed Abdelmotelb; Marzook, Fawzy Mohamed Abdelmaged [Atomic Energy Authority, Cairo (Egypt). Radioisotopes Production and Radioactive Sources Div.

    2016-08-01

    Levalbuterol was successfully radiolabeled with iodine using chloramine-T as an oxidizing agent via an electrophilic substitution reaction. The reaction parameters that affecting the labeling yield such as levalbuterol concentration, chloramine-T concentration, pH of the reaction medium and reaction time were studied in details. The radiochemical yield was 97.5 ± 0.5% and the radioiodinated compound was separated by HPLC. In vitro studies showed that the iodinated levalbuterol was stable for up to 24 h. The biodistribution in experimental animals showed that the lung uptake was 68.18 ± 0.17% at 5 min post injection which decreased with time until reached to 18.7 ± 0.12% at 2 h which was higher than other recent developed radiopharmaceuticals for lung imaging. The clearance pathways from the mice appear to proceed via both hepatobiliary and renal pathways. Predosing the mice with cold levalbuterol reduced the lung uptake to 20 ± 1.3% and further confirms the high specificity and selectivity of {sup 125}I-levalbuterol for the lung.

  7. Rapid and efficient radiosynthesis of [{sup 123}I]I-PK11195, a single photon emission computed tomography tracer for peripheral benzodiazepine receptors

    Energy Technology Data Exchange (ETDEWEB)

    Pimlott, Sally L. [Department of Clinical Physics, West of Scotland Radionuclide Dispensary, Western Infirmary, G11 6NT Glasgow (United Kingdom)], E-mail: s.pimlott@clinmed.gla.ac.uk; Stevenson, Louise [Department of Chemistry, WestCHEM, University of Glasgow, G12 8QQ Glasgow (United Kingdom); Wyper, David J. [Institute of Neurological Sciences, Southern General Hospital, G51 4TF Glasgow (United Kingdom); Sutherland, Andrew [Department of Chemistry, WestCHEM, University of Glasgow, G12 8QQ Glasgow (United Kingdom)

    2008-07-15

    Introduction: [{sup 123}I]I-PK11195 is a high-affinity single photon emission computed tomography radiotracer for peripheral benzodiazepine receptors that has previously been used to measure activated microglia and to assess neuroinflammation in the living human brain. This study investigates the radiosynthesis of [{sup 123}I]I-PK11195 in order to develop a rapid and efficient method that obtains [{sup 123}I]I-PK11195 with a high specific activity for in vivo animal and human imaging studies. Methods: The synthesis of [{sup 123}I]I-PK11195 was evaluated using a solid-state interhalogen exchange method and an electrophilic iododestannylation method, where bromine and trimethylstannyl derivatives were used as precursors, respectively. In the electrophilic iododestannylation method, the oxidants peracetic acid and chloramine-T were both investigated. Results: Electrophilic iododestannylation produced [{sup 123}I]I-PK11195 with a higher isolated radiochemical yield and a higher specific activity than achievable using the halogen exchange method investigated. Using chloramine-T as oxidant provided a rapid and efficient method of choice for the synthesis of [{sup 123}I]I-PK11195. Conclusions: [{sup 123}I]I-PK11195 has been successfully synthesized via a rapid and efficient electrophilic iododestannylation method, producing [{sup 123}I]I-PK11195 with a higher isolated radiochemical yield and a higher specific activity than previously achieved.

  8. Studies on human proinsulin.C-peptide radioimmunoassay method

    International Nuclear Information System (INIS)

    125I-labelled human.C-peptide was prepared by the chloramin T method, the enzymic method and the active ester method, respectively. Using respective 125I-labelled human.C-peptides in human proinsulin.C-peptide RIA, we compared the binding (B0/T %) to antibody, displacement by standard human.C-peptide, recovery, and stability. The usable 125I-labelled antigen for human proinsulin.C-peptide RIA could be prepared by the chloramin T method and the enzymic method which labelled 125I to tyrosyl human proinsulin connecting peptide, and by an active ester method which conjugates 125I-labelled active ester to human proinsulin connecting peptide. No differences among those 125I-labelled antigens were observed in displacement (B/B0 %) by standard human.C-peptide or the recovery test. In the case of constant preparation of 125I-labelled antigen for RIA, the enzymic method was the best from the viewpoint that reaction ratio is stable and the stability of B0/T % was good. (auth.)

  9. Preparation of 131I-epidepride and biodistribution study in SD rats

    International Nuclear Information System (INIS)

    The purpose is to study the preparation and biodistribution in animals of dopamine D2 receptor ligand 131I-epidepride. Epidepride was 131I radioiodinated with hydroperoxide or chloramine-T method. The purity and stability of 131I-epidepride were studied by HPLC and TLC, the dynamic distribution in brain and critical organs of SD rats were studied. The radiolabelling yields of 131I-epidepride were 97.4% and 52.9% respectively. With no more purification, the hydroperoxide method is better than the chloramine-T method. After 4 hours in room temperature, the radiochemical purity of 131I-epidepride was still over 90%. The striatum uptake was good, the striatum/cerebellum ratio reached 237:1 at 320 min. Among the critical organs, lung had the first and the highest uptake. All organs had quick clearance (T1/2131I-epidepride has a high striatum uptake and a high striatum/cerebellum ratio. 131I-epidepride is a good dopamine D2 receptor agent

  10. Optimization of the preparation conditions of radioiodoepidepride. A dopamine D2receptor antagonist radioligand

    International Nuclear Information System (INIS)

    [125I]iodepidepride, (s)-(-)-[(1-ethyl-2-pyrrolidinyl)methyl]-5-[125I]-iodo-2,3- dimethoxybenzamide is the iodine substituted analogue of isoremoxipride, both of which are very potent dopamine D2-antagonists. Epidepride was radioiodinated using different oxidizing agents such as chloramine-T, iodogen, iodogen glass frit and hydrogen peroxide. Chloramine-T is a powerful oxidizing agent compared to both iodogen and hydrogen peroxide so that the side products, especially the chlorinated epidepride, decreases the radiochemical yield. This chlorinated epidepride is minimized in the case of iodogen and iodogen glass frit and are not observed in case of the non-chlorinated oxidizing agent hydrogen peroxide. TLC and HPLC were used to analyze the reaction components and to estimate both the radiochemical yield and purity. The reaction parameters such as reaction time, pH, epidepride and oxidizing agent concentrations and the stabilty of the final product were studied to optimize the radiochemical yield and purity. The optimized radiochemical yield was about 90% and the radiochemical purity of the final product was 99.9%. (author)

  11. Labeling Lanreotide with 125I and 188Re

    International Nuclear Information System (INIS)

    Lanreotide is a new somatostatin analogue. It can bind to human somatostatin receptor (hSSTR) subtype 2 through 5 with high affinity and to hSSTR subtype I with low affinity. We investigate labeling condition, quality control and stability in vitro of 125I-Lanreotide and 188Re-lanreotide respectively. (A) Lanreotide is labeled with 125I using Chloramine T. The effect of reaction condition (such as reaction time, pH value, Lanreotide amount, quantity of Chloramine T and reaction volume) on labeling yield is investigated in detail. (B) The labeling yield and radiochemical purity (RP) is measured with paper chromatography (PC) and Sep-Pak C18 Cartridge. (C) The stability of 125I-Lanreotide in vitro is investigated by labeling compound incubating for 48 hours at 37 deg C in the 0.9% sodium chloride solution and RP is tested by PC at specific time intervals. (D) Lanreotide is labeled directly with 188Re via the mixture of citrate and tartate using stannous chloride as reduced agent. The influence of reaction conditions such as pH, temperature, amount of stannous chloride, amount of Lanreotide and reaction time on labeling yield is investigated in detail. At the time, the stability in vitro quality control and animal test are evaluated

  12. Clinical scale preparation and evaluation of 131I-Rituximab for Non-Hodgkin's lymphoma

    International Nuclear Information System (INIS)

    Radioimmunotherapy (RIT) with anti CD20 MoAb conjugated to a β- emitting radioisotope like 131I or 90Y has the added advantage of delivering radiation not only to tumor cells that bind the antibody but also due to a crossfire effect, to neighboring tumor cells inaccessible to the antibody. In order to make available an indigenous radioimmunotherapeutic agent for Non Hodgkin's Lymphoma (NHL), radioiodinated Rituximab has been prepared and evaluated at a clinical scale. Radioiodination of Rituximab was performed by the conventional Chloramine T method using 7.4 GBq Na131I in a lead shielded plant. Six batches of radioiodination were prepared and characterized by electrophoresis and HPLC to evaluate the reproducibility of the product. The product remained stable retaining the radiochemical purity > 95% upto 5 days after radioiodination. In vitro cell binding studies and biodistribution studies in normal Swiss mice have indicated the potential of this molecule as a radioimmunotherapeutic agent for NHL. (orig.)

  13. Standardization of human thyrotropin radioimmunoassay and its application to the purification of this hormone to the preparation of the assay reagents

    International Nuclear Information System (INIS)

    The various steps that are necessary for setting up the thyrotropin radioimmunoassay are presented below. Radioiodination was carried out through the Chloramine T method and the labeled purification performed on Sephadex G-100. Purification of human thyrotropin from side fractions obtained during the purification of growth hormone was carried out in order to obtain a pure reagent for use in the radioimmunoassay. The employment of the hormone obtained was evaluated as the radioimmunoassay tracer in comparison with that prepared from the hormone received from the NIDDKD, U.S.A. The results indicated that although it was not possible to obtain a hormone with a purity degree adequate to be used as the tracer, enough experience was acquired for the isolation of thyrotropin. (author)

  14. Study and development of a radioimmunoassay of antidiuretic hormone sensitive at 10/sup -12/M

    Energy Technology Data Exchange (ETDEWEB)

    Caillens, H.; Rousselet, F. (Faculte des Sciences Pharmaceutiques, (France)); Paillard, F. (Hopital Tenon, Paris (France))

    1982-07-01

    A radioimmunoassay of antidiuretic hormone is described. The antiserum was obtained by immunization of rabbits with lysine vasopressin conjugated to hemocyanine. The specificity of the antibody was selective and directed against the pentapeptide ring of the vasopressin molecule: oxytocin showed no cross-reactivity at 10/sup -9/M. The labelled hormone (/sup 125/I-AVP) prepared using the chloramine-T method had a high specific activity (1860 Ci/mmol). Incubation was performed in an equilibrium system. Comparative studies of different separation methods of bounds and free /sup 125/I-AVP showed that the sensitivity and the precision of the standard curve were better using charcoal dextran. The limit of detection of the assay was 1,6 pg per ml.

  15. Homologous radioimmunoassay of human prolactin

    International Nuclear Information System (INIS)

    Gelfiltration on Sephadex G-75 showed a heterogenity of prolactin in serum of patients with prolactinoma and in culture medium of a prolactinoma. Serum of patients with prolactinoma and culture medium of a prolactinoma were examined as possible sources of prolactin by gel filtration and ion exchange chromatography. Polyacrylamide electrophoresis revealed both preparations as contaminated by other proteins. Nevertheless prolactin isolated form culture medium of a prolactinoma is good enough as a tracer in our radioimmunoassay because contaminating proteins in this preparation do not inferfere in our system. An hPRL antiserum created in a rabbit against a crude fraction of human serum of a patient with prolactinoma was tested by titration, saturation studies, and ion exchange chromatography. In comparison with lactoperoxidase-iodinated prolactin Chloramine T iodinated prolactin showed higher loss of immunochemical properity, however higher specific activity. Specifity and precision in our radioimmunoassay system were described and the conditions of optimal sensitivity in our assay were evaluated. (orig.)

  16. Rapid method for the preparation of 125I-labelled human growth hormone for receptor studies, using reverse-phase high performance liquid chromatography

    International Nuclear Information System (INIS)

    Human growth hormone was labelled with 125 Iodine by the stoichiometric modification of the chloramine-T method to a specific activity of 50-80 microCi/microgram, and the iodinated mixture was purified by reverse-phase high performance liquid chromatography using a C18 column (SynChropak RP-P) and a linear gradient. Compared with the usual Sephadex G-100 chromatography, HPLC gave a much better separation, with a higher yield and a considerably reduced analysis time (30 min vs 5 h). The [125I]-labelled preparation had normal binding to IM-9 lymphocyte receptors. The maximum bindability of the HPLC-purified preparation approximated 90%, which is the best value so far reported for human growth hormone. It is concluded that HPLC is a fast, convenient and reproducible method for obtaining an improved [125I]-labelled human growth hormone for receptor studies

  17. Development of the kits for RIA simultaneous determination of polypeptide hormones

    International Nuclear Information System (INIS)

    A simple and universal modification of chloramine T technique has been developed for the radioactive iodination of several polypeptide hormones such as insulin, human growth hormone (HGH), human TSH, synthetic human gastrin and beta-endorphine. The prepared products proved to have good immunoreactivity suitable for RIA purposes. The technique is inexpensive and quick. A new procedure has also been worked out utilizing horse myeloperoxidase in solid state as catalyser. The hormones iodinated with this technique show better parameters (e.g. longer stability, better binding to antibody, more favourable adsorption on dextran-coated charcoal); however the specific activities achieved were lower. The possibilities of simultaneous measurement of insulin and HGH have been studied. In this connection, a comparatively simple method for the determination of the endogenous anti-insulin antibodies was developed and used for the control of patients with diabetes and for the checking of new insulin preparations. However, the technique requires relatively sophisticated equipment and computerized calculations

  18. Spectrophotometric determination of azathioprine in pharmaceutical formulations.

    Science.gov (United States)

    Lakshmi, C S; Reddy, M N

    1998-12-01

    Four simple and sensitive visible spectrophotometric methods (A-D) have been described for the assay of azathioprine (ATP) either in pure form or in pharmaceutical formulations. Methods A and B are based on the oxidation of ATP with excess N-bromosuccinimide (NBS) or chloramine-T (CAT) and determining the consumed NBS or CAT with a decrease in colour intensity of celestine blue (CB) (method A) or gallocyanine (GC) (method B), respectively. Methods C and D are based on the diazotisation of reduced azathioprine (RATP) with excess nitrous acid and estimating either the consumed nitrous acid (HNO(2)) with cresyl fast violet acetate (CFVA) (method C) or by coupling reaction of the diazonium salt formed with N-1-naphthyl ethylene diamine dihydrochloride (NED) (method D). All of the variables have been optimized and the reactions presented. The concentration measurements are reproducible within a relative standard deviation of 1.0%. Recoveries are 99.2-100.3%. PMID:18967434

  19. Invertase immobilization by adsorption on polymer microspheres studied by radioiodination technique

    Energy Technology Data Exchange (ETDEWEB)

    Queiroz, Alvaro A.A. de [Universidade Federal de Itajuba, MG (Brazil). Inst. de Ciencias Exatas. Dept. de Fisica e Quimica]. E-mail: alencar@unifei.edu.br; Pontin, Luiz F. [Universidade Federal de Itajuba, MG (Brazil). Inst. de Ciencias Exatas. Dept. de Matematica e Computacao]. E-mail: pontin@unifei.edu.br; Higa, Olga Z.; Ribela, Maria Tereza C.P. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Lab. de Biologia Molecular]. E-mail: ozahiga@ipen.br; Tomotani, Ester J.; Vitolo, Michele [Sao Paulo Univ., SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Tecnologia Bioquimico-Farmaceutica]. E-mail: michenzi@usp.br

    2005-07-01

    In this paper, we report the study of the diffusion behavior of invertase onto polystyrene-divinylbenzene (PS-DVB) microspheres. To detect the surface concentration of protein adsorbed on the microspheres, the chloramine-T method was used to label invertase and has been applied to the study of protein sorption properties on the microspheres. The sorption isotherms and the diffusion coefficient (D{sub f}) were computed from experimental results and the concentration of bound invertase was determined in terms of the Fick's law. The Hill equation was applied to the data and the binding capacity of the microspheres were estimated. The results of adsorption show that the radiolabelling invertase method are efficacious at the protein surface concentration detection and can be used to investigate the enzyme immobilization by sorption properties of polymer microspheres. (author)

  20. Development of glycoside-bound radiopharmaceuticals; Novel radioiodination method for digoxin

    Energy Technology Data Exchange (ETDEWEB)

    Takemura, Yasutaka; Dote, Nobuhito; Taniuchi, Hideyuki; Iijima, Naoko; Yokoyama, Akira (Kyoto Univ. (Japan). Faculty of Pharmaceutical Science); Fujibayashi, Yasuhisa; Konishi, Junji

    1994-01-01

    We combined 2-hydroxy-3-methylbenzoylhydrazide (HMBH) with glycosides as a novel method for the radioiodination of physiologically active glycosides. This method was tested using digoxin, which is one of the cardiac glycosides. A digoxin-HMBH conjugate was synthesized by periodate cleavage of the third sugar ring, and was readily radiolabelled with Na[[sup 125]I] by the chloramine-T method. [sup 125]I labelled digoxin-HMBH conjugate retained Na[sup +], K[sup +]-ATPase binding in vivo and in vitro, and also retained immunoreactivity to an anti-digoxin antibody. Thus, this [sup 125]I labelled digoxin-HMBH conjugate represents a potential radiopharmaceutical for Na[sup +], K[sup +]-ATPase imaging, as well as for the radioimmunoassay of digoxin. (author).

  1. A radioimmunoassay for chicken avidin

    International Nuclear Information System (INIS)

    A double-antibody solid-phase radioimmunoassay for chicken avidin is reported. Avidin was labelled with 125I by the chloramine-T method. The bound and free avidin were separated with a second antibody bound to a solid matrix. In the logit-log scale the standard curve was linear from 1-2 to 100-200ng of avidin/ml. Cross-reaction of ovalbumin was less than 0.015%. Saturation of biotin-binding sites of avidin with an excess of biotin decreased radioimmunoassay values by about 15%. Recovery studies indicated that avidin can be assayed from all chicken tissues studied with radioimmunoassay, whereas the [14C]biotin/bentonite method gave poor recoveries for avidin in the liver and kidney. Radioimmunoassay and the [14C]biotin/bentonite method gave similar concentrations for oviduct avidin. (author)

  2. Radioimmunoassay determination of decreased amounts of. cap alpha. -L-fucosidase protein in fucosidosis

    Energy Technology Data Exchange (ETDEWEB)

    Andrews-Smith, G.L.; Alhadeff, J.A. (California Univ., San Diego, La Jolla (USA). Dept. of Neurosciences)

    1982-03-15

    Purified human liver ..cap alpha..-L-fucosidase (EC 3.2.1.51) has been radioiodinated by a chloramine-T procedure to a specific activity of 3.7 x 10/sup 6/ dpm/..mu..g protein without altering its apparent Michaelis constant for the 4-methylumbelliferyl substrate. This /sup 125/I-labeled ..cap alpha..-L-fucosidase has been used in development of a competitive binding radioimmunoassay for ..cap alpha..-L-fucosidase which can detect 1-2 ng of enzyme protein and has been employed to quantify the amount of ..cap alpha..-L-fucosidase protein in the liver and spleen from a patient with fucosidosis. Less than 1% of the normal amount of ..cap alpha..-L-fucosidase protein is present suggesting that normal amounts of catalytically inactive ..cap alpha..-L-fucosidase are not found in this disease.

  3. Fungicidal effect of 15 disinfectants against 24 fungal contaminants commonly found in bread and cheese manufacturing

    DEFF Research Database (Denmark)

    Bundgaard-Nielsen, Kirsten; Nielsen, Per Væggemose

    1996-01-01

    Resistance of 19 mold- and 6 yeast- species against 15 commercial disinfectants was investigated by a suspension-method in which the fungicidal effect and germination time were determined at 20 °C. Disinfectants containing 0.5 % dodecyldiethylentriaminacetic acid, 10 g/l chloramine-T, 2.......0 % formaldehyde, 0.1 % potassium hydroxide, 3.0 % hydrogen peroxide or 0.3 % peracetic acid were ineffective as fungicides. The fungicidal effect of quaternary ammonium compounds and chlorine compounds showed great variance between species and among the 6 tested isolates of Penicillium roqueforti var. roqueforti...... with age. Both P. roqueforti var. roqueforti and E. repens showed great variability of resistance within isolates of each specie....

  4. Synthesis and evaluation of an (125)I-labeled azide prosthetic group for efficient and bioorthogonal radiolabeling of cyclooctyne-group containing molecules using copper-free click reaction.

    Science.gov (United States)

    Choi, Mi Hee; Shim, Ha Eun; Nam, You Ree; Kim, Hye Rim; Kang, Jung Ae; Lee, Dong-Eun; Park, Sang Hyun; Choi, Dae Seong; Jang, Beom-Su; Jeon, Jongho

    2016-02-01

    Herein we report the radiosynthesis of a pyridine derived azide prosthetic group for iodine radioisotope labeling of dibenzocyclooctyne (DBCO) conjugated molecules. The radiolabeling of the stannylated precursor 2 was conducted using [(125)I]NaI and chloramine-T to give (125)I-labeled azide ([(125)I]1) with high radiochemical yield (72±8%, n=4) and radiochemical purity (>99%). Using (125)I-labeled azide ([(125)I]1), cyclic RGD peptide and near infrared fluorescent molecule were efficiently labeled with modest to good radiochemical yields. The biodistribution study and SPECT/CT images showed that [(125)I]1 underwent rapid renal clearance. These results clearly demonstrated that [(125)I]1 could be used as an useful radiotracer for in vivo pre-targeted imaging as well as efficient in vitro radiolabeling of DBCO containing molecules. PMID:26748695

  5. Linking water treatment practices and fish welfare

    DEFF Research Database (Denmark)

    Zubiaurre, Claire; Pedersen, Lars-Flemming

    2016-01-01

    Peracetic acids can be used as sanitizers to control water quality in aquaculture systems. As an alternative to formalin, chloramine-T or copper sulphate, PAA has strong anti-microbial effects, degrades quickly and is relatively safe to use. Its mode of action and associated rapid decay can make....... Supportive enzymatic, biochemical and physiological biomarkers can be used along with gill and epidermal histological measures to evaluate the effects on water treatment regimens. The ultimate goal is to define the therapeutic window where fish welfare is not compromised.PAA is among the few disinfectants...... optimizing treatment protocols a challenge. Continuous low-dose applications seem to be a promising solution. In this preliminary study behavioral response was used to assess potential correlations with PAA dosage. A behavioral change or response is not necessarily an indication of compromised welfare...

  6. Development and Optimization of Sepharose Solid Phase Radioimmunoassay for Estimation of Thyroid Stimulating Hormone

    International Nuclear Information System (INIS)

    The aim of the present study was oriented to develop, optimize and validate solid phase radioimmunoassay, through many studies on Sepharose, for estimation of thyroid stimulating hormone in humane serum. Preparation of polyclonal antibodies was carried out in host rabbit animals against TSH antigen followed by partial purification of 1gG. Linkage of antibody 1gG to activated Sepharose CL-4B was carried out after activation of Sepharose with 1,1- carbonyldiimidazole. Labeling of TSH was carried out using chloramine-T as an oxidizing agent and the labeled tracer was purified through PD-10 column. Extensive studies were carried out to obtain the optimum conditions of using solid phase Sepharose to reach higher separation efficiency. The results of validation tests reveal that the local solid phase system is precise and accurate for evaluation of thyroid disorders

  7. Preparation and Validation of Double Antibody Radioimmunoassay for Thyroglobulin (Tg) using Balb/C Mice as Host Animals

    International Nuclear Information System (INIS)

    The preparation and development of primary reagents of thyroglobulin (Tg) radioimmunoassay technique with low cost is considered to be the main objective of present study. The production of polyclonal antibodies of thyroglobulin was undertaken by immunizing three bulb/C mice intraperitoneal through primary injection and two booster doses. The preparation of 125I-Tg radiotracer was carried out using chloramine-T as oxidizing agent. The preparation of thyroglobulin standards were carried out. Optimization and validation of the assay were studied out. The results obtained provide a highly sensitive, specific and accurate RIA system for thyroglobulin based on liquid phase separation. In conclusion, this assay could be used for diagnosis of thyroid cancer.

  8. Iodination of monoclonal antibodies, proteins and peptide using iodogen

    International Nuclear Information System (INIS)

    The use of the iodinating reagent 1,3,4,6-tetrachloro-3α, 6α-diphenylglycholuril (Iodogen) to label monoclonal antibodies (McAbs). Proteins and peptides was invesrigated with McAbs identified as mouse IgG and IgM, arginine-vasopressin (AVP), glucagon (Glu), human insulin(hI) and albumin(Alb). The labeled products were purified by gel chromatography and their immunoreactivity were detected by RIA or IRMA> Comparison of the Iodogen method with the lactoperoxides and chloramine-T methods showed that the Iodogen method had a number of advantages: 1) technically simpler ; 2) a high labeling efficiency could be obtained; 3) the immunoreactivity of the products was minimally affected; 4) the products were stable for up to 4 months

  9. Standardisation of radioimmunoassay for human insulin employing magnetizable cellulose particles

    International Nuclear Information System (INIS)

    We describe a convenient and flexible solid phase radioimmunoassay for human insulin employing magnetizable cellulose particles. Anti-porcine insulin antibody was covalently linked to magnetizable cellulose particles to form a stable and economical solid phase immunosorbent system. The tracer was prepared by radioiodinating insulin with 125I using Chloramine-T oxidation method. The analytical sensitivity of assay observed was 5.5 μIU/mL. Intra-assay and inter-assay variations were found to be <12 % along with analytical recovery of 93-109 %. The developed assay can be used for the routine analysis of clinical samples. In addition, concentration of the solid phase magnetizable immunosorbent can be easily varied as per the specific requirement for research purposes. (author)

  10. Measurement of plasma canine C peptide by radioimmunoassay

    International Nuclear Information System (INIS)

    A sensitive radioimmunoassay for canine C peptide (CCP) was established using synthetic CCP, a specific antiserum, and rabbit anti-guinea pig serum. Radioiodination was performed according to a modified chloramine T method. Tracer preparations have been used for 6 weeks after iodination. The standard curve ranges from 0.028 to 3.0 nmol/l. The intra-assay coefficient of variation (CV) was 3-5% and the inter-assay CV was 6-9% in the optimal range between 0.3 and 0.8 nmol/l. The average recovery of CCP added to plasma samples was 100.6% (n = 9). Canine insulin, porcine proinsulin, bovine proinsulin, and human C peptide exhibited no cross-reactivity. The mean fasting plasma CCP concentration was 0.089 +- 0.021 nmol/l in normal dogs and -0.005 +- 0.007 nmol/l (mean +- SEM) in diabetic dogs, respectively. (author)

  11. Fetal antigen 2: an amniotic protein identified as the aminopropeptide of the alpha 1 chain of human procollagen type I

    DEFF Research Database (Denmark)

    Teisner, B; Rasmussen, H B; Højrup, P;

    1992-01-01

    -PAGE analysis gave an M(r) = 27 kDa under reducing and non-reducing conditions for both forms, whereas the exact M(r) determined by mass spectrometry was 14,343 +/- 3 Da. FA2 was N-terminally blocked and after tryptic digestion the amino acid composition and sequences of the peptides showed identity...... with the aminopropeptide of the alpha 1 chain of human procollagen type I as determined by nucleotide sequences. After oxidative procedures normally employed for radio-iodination (iodogen and chloramine-T), FA2 lost its immunoreactivity. An antigen which cross-reacted with polyclonal rabbit anti-human FA2 was demonstrated...... in fetal calf serum. Gel filtration with analysis of fractions by inhibition ELISA showed that the bovine homologue was present in the same molecular forms as those in human amniotic fluid, and immunohistochemical analysis with anti-human FA2 showed that its distribution in bovine skin was identical...

  12. UP-REGULATION OF HEPATIC RECEPTOR FOR GROWTH HORMONE IN THE FLOUNDER (PARALICHTHYS OLIVACEUS) AFTER ORAL ADMINISTRATION WITH EXOGENOUS GH

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The iodination efficiency of salmon GH(Sgh) was 38.82%,using a modification of the chloramine-T method. The specific activity of the 125I-Sgh was about 40 μCi/μg protein. The results of binding assay showed a single class of high affinity and low-capacity binding site in flounder liver. Long-term administration with exogenous GH can induce the up-regulation of hepatic GH receptor in total binding capacity though there was no significant difference of association constant among any groups. Considering that there was no significant difference in capacity of free binding sites of livers from control and experimental fish, this result also indicated that the liver from experimental fish, compared to that from control fish, had more occupied binding sites.

  13. Preparation of [123I]- and [125I]epidepride: a dopamine D-2 receptor antagonist radioligand

    International Nuclear Information System (INIS)

    (S)-(-)-N-[(1-ethyl-2-pyrrolidinyl)methyl]-5-[123I] iodo-2,3-dimethoxybenzamide (TDP 517) (proposed generic name, [123I]epidepride) is the iodine-123 substituted analogue of isoremoxipride (FLB 457), both of which are very potent dopamine D-2 antagonists (epidepride KD 0.024 nM). [123I] Epidepride was radioiodinated in 60-70% radiochemical yields in 35 min from the corresponding 5-(tributyltin) derivative using Na123I with a specific radioactivity of 3000 Ci/mmol, and oxidized in situ with chloramine-T. The aryltin precursor was prepared from non-labelled epidepride by palladium-catalyzed stannylation using bis(tri-n-butyltin) in triethylamine. Alternatively, using no carrier-added Na125I as the radioisotope, [125I] epidepride at 2000 Ci/mmol specific radioactivity was prepared in 86% radiochemical yield and 99% radiochemical purity after purification by reverse phase HPLC in ethanolic phosphate buffer. (author)

  14. UP-REGULATION OF HEPATIC RECEPTOR FOR GROWTH HORMONE IN THE FLOUNDER (PARALICHTHYS OLIVACEUS) AFTER ORAL ADMINISTRATION WITH EXOGENOUS GH

    Institute of Scientific and Technical Information of China (English)

    刘宗柱; 王金宝; 徐永立; 王勇; 张培军

    2001-01-01

    The iodination efficiency of salmon GH (sGH) was 38.82%,using a modification of the chloramine-T method. The specific activity of the 125I-sGH was about 40 μCi/μg protein. The results of binding assay showed a single class of high affinity and low-capacity binding site in flounder liver. Long-term administration with exogenous GH can induce the up-regulation of hepatic GH receptor in total binding capacity though there was no significant difference of association constant among any groups. Con-sidering that there was no significant difference in capacity of free binding sites of livers from control and experimental fish, this result also indicated that the liver from experimental fish, compared to that from control fish, had more occupied binding sites.

  15. Receptor-purified, Bolton-Hunter radioiodinated, recombinant, human epidermal growth factor: An improved radioligand for receptor studies

    International Nuclear Information System (INIS)

    We report an assessment of the applicability of the Bolton-Hunter method to the radioiodination of epidermal growth factor (EGF). Recombinant human EGF (hEGF) could be radioiodinated successfully by this method, whereas murine EGF could not. Bolton-Hunter 125I-labeled hEGF was compared with commercial 125I-labeled hEGF prepared by the chloramine-T radioiodination method. Neither radioligand was sufficiently pure for a detailed characterization of the purportedly heterogeneous pattern of binding of EGF to its receptors. A procedure based on receptor adsorption was thus developed for repurification of the Bolton-Hunter 125I-labeled hEGF. This provided a much purer radioligand suitable for detailed studies of receptor-binding heterogeneity

  16. Iodination of Fab fragments: Effect of I/Fab molar ratio

    Energy Technology Data Exchange (ETDEWEB)

    Kishore, R.; Eary, J.F.; Beaumier, P.L.; Hellstrom, K.E.; Hellstrom, I.; Nelp, W.B.

    1985-05-01

    Radioisotopes of iodine covalently coupled to antibodies have formed the standard against which other radiolabeled antibody tracers are compared. Since the use of monoclonal antibodies (MoAb) for the diagnosis and therapy of malignant diseases is increasing rapidly the authors wished to investigate radiolabeling variables which affect the immunointegrity of radioiodinated antibodies. The authors studied the electrophilic (chloramine-T) iodination of the Fab fragment of a murine MoAb against the high molecular weight proteoglycan antigen of human melanoma. The immunoreactivity of the iodianted Fab was assessed using a cell binding assay with formaldehyde-fixed cells of a selected cell line (number 2669). The in vitro stability of the labeled fragment was studied electrophoretically. The results indicate that the reaction time and concentrations of chloramine-T were not critical within broad limits. On the other hand immunoreactivity and deiodination over time (shelf-life) were inversely related and very sensitive to I/Fab molar ratio even at concentrations below -0.1 atom of I per Fab molecule. This has important implications for the radiotherapy of malignant tumors using I-131 labeled immunoglobulins which often have higher I/Fab molar ratios (100 mCi I-131/10 mg Fab approx. = 0.5 I atoms/Fab) versus diagnostic preparations (10 mCi I-131/5 mg Fab approx. = 0.1 I atoms/Fab). Thus the authors conclude that to maintain high immunointegrity the I/Fab molar ratio should be kept low, especially for therapeutic preparations, by using correspondingly higher amounts of Fab.

  17. Generation of a high-valent iron imido corrolazine complex and NR group transfer reactivity.

    Science.gov (United States)

    Leeladee, Pannee; Jameson, Guy N L; Siegler, Maxime A; Kumar, Devesh; de Visser, Sam P; Goldberg, David P

    2013-04-15

    The generation of a new high-valent iron terminal imido complex prepared with a corrolazine macrocycle is reported. The reaction of [Fe(III)(TBP8Cz)] (TBP8Cz = octakis(4-tert-butylphenyl)corrolazinato) with the commercially available chloramine-T (Na(+)TsNCl(-)) leads to oxidative N-tosyl transfer to afford [Fe(IV)(TBP8Cz(+•))(NTs)] in dichloromethane/acetonitrile at room temperature. This complex was characterized by UV-vis, Mössbauer (δ = -0.05 mm s(-1), ΔE(Q) = 2.94 mm s(-1)), and EPR (X-band (15 K), g = 2.10, 2.00) spectroscopies, and together with reactivity patterns and DFT calculations has been established as an iron(IV) species antiferromagnetically coupled with a Cz-π-cation-radical (S(total) = 1/2 ground state). Reactivity studies with triphenylphosphine as substrate show that [Fe(IV)(TBP8Cz(+•))(NTs)] is an efficient NTs transfer agent, affording the phospharane product Ph3P═NTs under both stoichiometric and catalytic conditions. Kinetic analysis of this reaction supports a bimolecular NTs transfer mechanism with rate constant of 70(15) M(-1) s(-1). These data indicate that [Fe(IV)(TBP8Cz(+•))(NTs)] reacts about 100 times faster than analogous Mn terminal arylimido corrole analogues. It was found that two products crystallize from the same reaction mixture of Fe(III)(TBP8Cz) + chloramine-T + PPh3, [Fe(IV)(TBP8Cz)(NPPh3)] and [Fe(III)(TBP8Cz)(OPPh3)], which were definitively characterized by X-ray crystallography. The sequential production of Ph3P═NTs, Ph3P═NH, and Ph3P═O was observed by (31)P NMR spectroscopy and led to a proposed mechanism that accounts for all of the observed products. The latter Fe(III) complex was then rationally synthesized and structurally characterized from Fe(III)(TBP8Cz) and OPPh3, providing an important benchmark compound for spectroscopic studies. A combination of Mössbauer and EPR spectroscopies led to the characterization of both intermediate spin (S = 3/2 and low spin (S = 1/2) Fe(III) corrolazines, as

  18. Labeling of Octreotide With 131Ⅰ and Its Vitro Uptake in Inflammatory Cells for Graves' Ophthalmopathy%Octreotide的131I标记及Graves眼病炎症细胞的体外摄取

    Institute of Scientific and Technical Information of China (English)

    卢霞; 王荣福; 闫平; 周颖明; 张一帆

    2011-01-01

    将octreotide中的苯丙氨酸用酪氨酸替换合成[Tyr3]octreotide,对其进行了131I标记.考察了氯胺T标记法中氯胺T和[Tyr3]octreotide的用量对标记率的影响,观察了Graves眼病相关炎症细胞对131I-[Tyr3]octrotide的体外摄取情况.结果表明,对octreotide进行结构修饰后得到的[Tyr3]octreotide可以用于131I标记;氯胺T法对[Tyr3]octreotide进行131I标记时,氯胺T和[Tyr3]octreotide的最佳用量分别为1.6和1.4 μg,最高标记率为85%;T淋巴细胞、活化的成纤维细胞及血管内皮细胞均能够摄取131-[Tyr3]octreotide,而脂肪细胞的摄取则明显较少.以上结果表明,131I-[Tyr3]octreotide能够被Graves眼病发病过程中的炎症细胞摄取,具有用于Graves眼病发病过程中炎症反应程度评估的应用潜力.但由于摄取量还很小,需要进一步深入研究.%Phenylalanine was displaced by tyrosine in octreotide([Tyr3]octreotide) to study the labeling methods of [Tyr3]octreotide with 131Ⅰ, and the difference uptake ability of 131 Ⅰ-[Tyr3]octreotide in various cell lines of Graves ophthalmopathy were observed. The results showed that [Tyr3]octreotide could be labeled with 131Ⅰ by chloramines-T method and the optimal conditiona was 1.6μg chloramines-T and 1.4μg [Tyr3]octreotide. The labeling efficiency was 85%. T lymphocyte, fibroblast, and HUVEC could accumulate 131 Ⅰ-[Tyr3]octreotide largely compared to lipocyte, which indicated that 131Ⅰ-[Tyr3]octreotide could be combined with inflammatory cells in Graves' ophthalmopathy.

  19. Labeling Lanreotide with 125I and 188Re. China

    International Nuclear Information System (INIS)

    Lanreotide (D-β-Nal-Cys-Try-D-Trp-Lys-Val-Cys-Thr-NH2) is a new somatostatin analogue. It can bind to human somatostatin receptor (hSSTR) subtype 2 through 5 with high affinity and to hSSTR subtype 1 with low affinity. We investigate labeling condition, quality control and stability in vitro of 125I-Lanreotide and 188Re-lanreotide respectively. (A) Lanreotide is labeled with 125I using Chloramine T. The effect of reaction condition (such as reaction time, pH value, Lanreotide amount, quantity of Chloramine T and reaction volume) on labeling yield is investigated in detail. (B) The labeling yield and radiochemical purity (RP) is measured with paper chromatography (PC) and Sep-Pak C18 Cartridge. For PC method, 125I-Lanreotide is spotted on the Whatman No.1 paper and developed in the mixture of CH3CH2CH2CH2OH and CH3CH2OH and NH4OH (v/v/v=5:2:1), the Rf value of every component in the mobile phase is given in table 1. For Sep-Pak C18 Cartridge methods each cartridge is washed with 10 ml of ethanol followed by 10 ml of iso-CH3CH2CH2OH solution. Aliquots of 0.1 mI sample is loaded onto the cartridge, unbound peptide (sodium iodine-125) is eluted with 5 ml of 0.5mol/L sodium acetate solution, 125I-Lanreotide is eluted with 5 mI of 95% aqueous ethanol solution. (C) The stability of 125I-Lanreotide in vitro is investigated by labeling compound incubating for 48 hours at 37 deg. C in the 0.9% sodium chloride solution and RP is tested by PC at specific time intervals. (D) Lanreotide is labeled directly with 188Re via the mixture of citrate and tartate using stannous chloride as reduced agent. The influence of reaction conditions such as pH, temperature, amount of stannous chloride, amount of Lanreotide and reaction time on labeling yield is investigated in detail. At the time, the stability in vitro quality control and animal test are evaluated

  20. Iodine-125 Chitosan-Vitamin C complex. Preparation, characterization and application

    Energy Technology Data Exchange (ETDEWEB)

    Elbarbary, Ahmed M. [National Center for Radiation Research and Technology, Cairo (Egypt). Polymer Chemistry Dept.; Shafik, H.M.; Ebeid, N.H.; Ayoub, S.M. [Atomic Energy Authority, Cairo (Egypt). Hot Lab. Center; Othman, Sameh H. [Atomic Energy Authority, Cairo (Egypt). Nuclear Research Center

    2015-07-01

    In heterogeneous conditions, water soluble Chitosan-Vitamin C Complex (CSVC) is successfully synthesized via the ionic interaction between γ-degraded CS and VC. Chitosan (CS) of low molecular weight (MW) is prepared using γ-irradiation method. The coupling of CS and vitamin C (VC) is carried out by the chemical treatment of VC with the γ-degraded CS. The formation of CSVC complex instead of physical mixture is confirmed by FT-IR and UV spectrometry. Characterization by transmission electron microscope (TEM) and dynamic light scattering (DLS) shows the formation of a nanostructure in 40 nm range. The preparation of labeled CSVC was performed using chloramines-T oxidation method. The labeling feasibility of CSVC nanostructure by Iodine-125 ({sup 125}I) is investigated. The optimized conditions of labeling are thought to be 50 μg of oxidizing agent, pH 3, and one minute reaction time. The Biodistribution activity of {sup 125}I radiolabeled CSVC nanostructure ({sup 125}I-CSVC) is examined on a group of different ascites tumor bearing mice. Calculation of the biodistribution percentages shows that the tumor, liver, and kidney are the targeting organs of {sup 125}I-CSVC nanostructure.

  1. Preparation, purification and primary bioevaluation of radioiodinated ofloxacin. An imaging agent

    Energy Technology Data Exchange (ETDEWEB)

    Kandil, Shaban; Seddik, Usama; Hussien, Hiba; Shaltot, Mohamed [Atomic Energy Authority, Cairo (Egypt). Cyclotron Project; El-Tabl, Abdou [Monofia Univ. (Egypt). Faculty of Science

    2015-07-01

    The broad-spectrum antibiotic agents have been demonstrated as promising diagnostic tools for early detection of infectious lesions. We set out ofloxacin (Oflo), a second-generation fluoroquinolone, for the radioiodination process. In particular, this was carried out with {sup 125}I via an electrophilic substitution reaction. The radiochemical yield was influenced by different factors; drug concentration, different oxidizing agents, e.g. chloramine-T, iodogen and n-bromosuccinimide, pH of medium, reaction time, temperature and different organic media. These parameters were studied to optimize the best conditions for labeling with ofloxacin. We found that radiolabeling in ethanol medium showed a 70% radiochemical yield of {sup 125}I-ofloxacin. The radioiodination was determined by means of TLC and HPLC. The cold labeled Oflo ({sup 127}I-Oflo) was prepared and controlled by HPLC. The cold labeled Oflo was also confirmed by NMR and MS techniques. Furthermore, biodistribution studies for labeled {sup 125}I-Oflo were examined in two independent groups (3 mice in each one); control and E. Coli-injected (inflamed). The radiotracer showed a good localization in muscle of thigh for inflamed group as compared to control. In conclusion, ofloxacine might be a promising target as an anti-inflammatory imaging agent.

  2. Development of an 'In vitro' system for the caption essay of T3; Desarrollo de un sistema 'In vitro' para el ensayo de captacion de T3

    Energy Technology Data Exchange (ETDEWEB)

    Lavalley, C.; Ferro, G.; Zambrano, F.; Lezama, J

    1990-02-15

    Triiodothyronine uptake (T3U) is a qualitative technique for evaluation of the unsaturation capacity of thyroid binding globulin (TBG). This paper presents results related to a T3 standardized serum and the integration of T3-I-125, and adsorbent for labelled hormone. Labelled hormone were prepared by the chloramine T method and then purified by high performance liquid chromatography. The specific activity was 500 {mu} Ci/ {mu} g. Various adsorbents such as: Norit A Charcoal, calcium silicate, talc, bovine serum albumin macroaggregated (BSAM) were used in different buffers as: Tris-HCl, barbital and Michaelis. Standardized serum was prepared by mixing different euthyroid sera. Best conditions for T3U assays were achieved with 15 mg/ml. BSAM at pH 8.6 in presence of Tris-HCl buffer for hypothyroid and hyperthyroid sera, for which we obtained < 0.9 {+-} 0.04 and > 1.1 {+-} 0.05 respectively as a T3U index with a 3.0 % of coefficient variation. The reagents so prepared can be conveniently used for T3U assays. (Author)

  3. Application of different /sup 125/I tracers in radioimmunoassays of estradiol-17. beta

    Energy Technology Data Exchange (ETDEWEB)

    Bienert, R.; Flentje, H.; Herzmann, H. (Akademie der Wissenschaften der DDR, Berlin-Buch. Zentralinstitut fuer Isotopen- und Strahlenforschung; Akademie der Wissenschaften der DDR, Leipzig. Zentralinstitut fuer Isotopen- und Strahlenforschung)

    1984-01-01

    Some different /sup 125/I-labelled estradiol tracers were produced by direct radioiodizing of estradiol and also of the histamine and tyramine conjugates of estradiol-3-carboxymethylether (E/sub 2/-3-CM) by means of the chloramine-T method. The linkage properties of these tracers were investigated in relation to the /sup 3/H-labelled estradiol opposite to the antisera, which were produced against the cow serum albumin (RSA) conjugates of E/sub 2/-3-CM and estradiol-6-carboxymethyloxime (E/sub 2/-6-CMO). As suitable system for the radioimmunological estradiol determination could be revealed 4-/sup 125/I-iodine estradiol in connection with one antiserum in each case of the radioligand antiserum combinations against E/sub 2/-3-CM-RSA- and E/sub 2/-6-CMO-RSA-conjugate. The double antibody method is used for separation in optimized RIA systems. The first and the second antibody reaction take place simultaneously.

  4. Preparation of directly iodinated steroid hormones and related directly halogenated compounds

    International Nuclear Information System (INIS)

    The preparation of directly iodinated radioactive steroid hormones is described for use in radioimmunoassays or radiolocalization and treatment of human breast tumours. The radioactive iodinated steroid hormone is prepared by reacting a parent steroid hormone with an alkali metal iodide containing radioactive 123I, 125I, 130I or 131I in the presence of hydrogen peroxide or chloramine-T. The parent steroid hormones include the adrenal corticosteroids, the estrogens, the progestogens, the progestins and the diuretic and antidiuretic agents. The radioactive iodinated steroid hormone is prepared by iodinating the parent steroid hormone directly on the cyclopentanophenanthrene nucleus. The radioactive iodinated steroid hormones have the same antigenicity and receptor site specificity as the parent steroid hormone. The invention is illustrated by 1) the method of iodination of estradiol-17β, 2) results for the percentage labelling of several steroids and steroid hormones, 3) results for the radioimmunoassay of 125I-estradiol and 4) results for the binding of directly iodinated estradiol-17β in an estrogen receptor assay of human breast cancer. (U.K.)

  5. Biodistribution and radioimmunoimage of iodine-125 labeled anti-human ADAM15 polyclonal antibody in nude mice bearing human gastric carcinoma xenografts

    International Nuclear Information System (INIS)

    Objective: To investigate the biodistribution of Iodine-125 labeled anti-human ADAM15 polyclonal antibody in nude mice bearing human gastric carcinoma xenografts. Methods: The anti-human ADAM15 polyclonal antibody was labeled with I-125 using Chloramine-T method. The labeling efficiency and radiochemical purity of 125I-anti-ADAM15 antibody were measured. The SPECT planar imaging of nude mice bearing gastric carcinoma xenografts were performed at 1, 4, 8, 24, and 48 h post-injection and the biodistribution of 125I-anti-ADAM15 antibody was measured at 48 h after injection. Results: The labeling efficiency of 125I-anti-ADAM15 antibody was (75.16±9.43)% and its radiochemical purity was (99.44±0.21)% . Tumors could be cleanly visualized in SPECT planar images, and the radioactivity ratio of tumor to non-tumor tissue was 3.84±0.43 at 48 h post-injection. Conclusion: 125I-anti-ADAM15 antibody can target the gastric carcinoma in vivo, and provide good radioimmunoimages. (authors)

  6. Experimental research for tumor VIP receptor imaging

    International Nuclear Information System (INIS)

    To study the possibility of radioactive labelled vasoactive intestinal peptide (VIP) for tumor VIP receptor imaging. 125I-VIP was prepared by chloramine-T method, and purified by Sephadex G-50 column chromatography. The bioactivity and stability of 125I-VIP were measured by silica 60 F254 TLC and competition test to SGC7901 cell in vitro. The biodistribution of 125I-VIP was studied in the nude mice bearing tumor. The results showed that labelled rate of 125I was 73.8%, the specific activity was 18.2 PBq/mol, the radiochemical purity (RCP) was over 98% and remained 96.3% after 48 days stored at -80 degree C. The specific binding of 125I-VIP to the SGC7901 cell was inhibited by VIP in dose dependence in the competition experiment. The radioactivity of tumor was higher than that of muscles in all phases (P<0.05-0.01), the peak activity of tumor occurred at 30 min (3.58 +- 0.48ID%/g) and the peak ratio of T/N occurred at 60 min after the injection. The activity of lungs was obviously higher than that of blood, the intestine was always in low level. Most of the activity in the body was mainly eliminated from kidney. The present study demonstrated that the radioactive labelled VIP is a promising agent for tumor VIP receptor scintigraphy

  7. Production of antibodies against secretin and their use for radioimmunoassay of secretin

    International Nuclear Information System (INIS)

    Synthetic thyroglobulin was bonded to bovine albunia and thyroglobulin according to the principle of the carbodiimide condensation reaction. 16 rabbits were immunized with these conjugates and with unconjugated secretin. Secretin labelling with 125I was carried out by the chloramin-T method. The tracer has a specific activity of 15.45 mCi/mg. A secretin RIA was developed using the double antibody method. The sensitivity of the system could be raised by variation of the specific activity of the tracer and optimisation of the incubation parameters. Antisera were compared. The titers of secretin/bovine albumine conjugate antisera were similar to the antisera against secretin thyroglobulin conjugate. The sensitivity of the standard curves was higher for secretin/bovine albumin conjugate antisera than for thyroglobulin conjugate antisera. Two antisera were tested for specificity. The detection threshold of antiserum S 5 IX was 12,43 pmol/l while the 50% intercept was at 55.45 pmol/l. This antiserum is particularly suitable for a secretin RIA. (orig./MS)

  8. Preparation of 125I-labeled human growth hormone of high quality binding properties endowed with long-term stability

    International Nuclear Information System (INIS)

    125I-labeled human growth hormone (125I-labeled.hGH) was prepared by using two variants of the chloramine T labelling procedure and purified by polyacrylamide gel electrophoresis (PAGE) of the reaction mixture. Variant A produced a tracer with high specific activity (100 +/- 10 microCi/microgram), high maximal binding capacity to antibodies (93%) and long-term stability (at least 150 days at -20/degree/C). No diiodinated tyrosil residues could be detected in this tracer. Variant B was devised to obtain higher yields of labeled hormone. The electrophoresis of the iodination mixture revealed two radioactive components with Rm values of 0.49 and 0.55 which result from the iodination of hGH variants preexisting in the starting material. Both tracers had similar specific activities (70 +/- 10 microCi/microgram), high maximal binding capacity to antibodies or receptors (80-100%, after 80 days of their obtention) and high stability (at least 100 days at -20/degree/C). It is concluded that the iododerivatives of hGH obtained by either method are adequate to perform radioimmunoassay and receptor studies and have long-term stability

  9. Iodine-123 labeled derivatives of methylphenidate: potential SPECT radiopharmaceuticals for brain dopamine transporters

    International Nuclear Information System (INIS)

    Since dl-threo-[11C]methylphenidate (Ritalin) and especially the more active enantiomer, d-threo-[11C]methylphenidate, have favorable properties for PET studies, we prepared two radioiodinated analogs of methylphenidate, p-[123I]iodomethylphenidate and m-[123I]iodo-p-hydroxymethylphenidate with a view to evaluating them as potential SPECT tracers. To prepare p-[123I]iodomethylphenidate, the p-tributyltin derivative was prepared from the previously reported p-bromomethylphenidate and reacted under acidic conditions with I-123 iodide plus chloramine-T at room temperature for 90 seconds. The predomimant radioactive product was obtained in 85% radiochemical yield and > 10 Ci/μmol specific radioactivity after HPLC purification. It had the same HPLC retention time as a spectroscopically characterized non-radioactive p-iodomethylphenidate standard prepared via nitration of methylphenidate and diazotization, after protection of the secondary amino group by benzoylation. A second radioiodinated methylphenidate derivative, m-[123)I]iodop-hydroxymethylphenidate was prepared in 80% radiochemical yield by direct iodination of the known p-hydroxymethylphenidate. In this case the non-radioactive standard was prepared by iodination of p-hydroxyritalinic acid using I2 and iodic acid, followed by esterification. (author)

  10. Iodine-125 Chitosan-Vitamin C complex. Preparation, characterization and application

    International Nuclear Information System (INIS)

    In heterogeneous conditions, water soluble Chitosan-Vitamin C Complex (CSVC) is successfully synthesized via the ionic interaction between γ-degraded CS and VC. Chitosan (CS) of low molecular weight (MW) is prepared using γ-irradiation method. The coupling of CS and vitamin C (VC) is carried out by the chemical treatment of VC with the γ-degraded CS. The formation of CSVC complex instead of physical mixture is confirmed by FT-IR and UV spectrometry. Characterization by transmission electron microscope (TEM) and dynamic light scattering (DLS) shows the formation of a nanostructure in 40 nm range. The preparation of labeled CSVC was performed using chloramines-T oxidation method. The labeling feasibility of CSVC nanostructure by Iodine-125 (125I) is investigated. The optimized conditions of labeling are thought to be 50 μg of oxidizing agent, pH 3, and one minute reaction time. The Biodistribution activity of 125I radiolabeled CSVC nanostructure (125I-CSVC) is examined on a group of different ascites tumor bearing mice. Calculation of the biodistribution percentages shows that the tumor, liver, and kidney are the targeting organs of 125I-CSVC nanostructure.

  11. Development and clinical application of human gastrin radioimmunoassay

    International Nuclear Information System (INIS)

    The determination of human gastrin levels in the blood is very important for diagnosis of gastrointestinal disorders. This work describes the radioimmunoassay of gastrin developed according to Russell et al. and its clinical application measuring fasting levels of this hormone in normal subjects, gastrectomized, chagasics, patients with chronic renal failure (CRF), pernicious anemia (PA) and Zollinger-Ellison syndrome (ZES). Synthetic human gastrin was used for radioiodination and as standard, while the specific antibody was raised in rabbits. Gastrin was radioiodinated by a modification of the chloramine T technique and purified by anion exchange chromatography in QAE-Sephadex A-25 to a specific activity around 200 uCi/ug. The assays were performed by incubation of 125I-gastrin, standard gastrin (zero to 500 pmol/l) or unknown samples with the antiserum for 4 days at 40C. The antibody bound and free 125 I-gastrin was separated by adsorption of the latter to the charcoal. The basal gastrin values of normal subjects ranged from 2 to 74 pmol/l, being these levels higher in the chagasics (from 6 to 261 pmol/l). Higher levels of gastrin were determined in patients with CRF (from 12 to 350 pmol/l), PA (from 160 to 680 pmol/l) and with ZES(1010 pmol/l), while very low levels were confirmed in gastrectomized (from 1 to 8 pmol/l). (author)

  12. Comparison of three radioligands, selenium-75, iodine-125, and tritium, in the radioimmunoassay of methotrexate

    International Nuclear Information System (INIS)

    Radioimmunoassays for methotrexate are described, involving use of a rabbit antiserum to a conjugate of the drug and bovine serum albumin and the drug labeled with tritium, selenium-75, or iodine-125. Of the two gamma emitters, the 75Se-labeled drug was prepared by the Radiochemical Centre, Amersham, England and the 125I-labeled drug in the laboratory, by the Chloramine T iodination technique. The stability of labels with both methods allows use of the faster, cheaper, and simpler gamma-counting techniques, with results available after 3 h. All three methods have acceptable sensitivity, accuracy, precision, and reproducibility, and are specific for methotrexate, with no significant interference from naturally occurring folates or leucovorin. The assays in which the gamma emitters are used have significant practical advantages over the beta emitter and are much better suited to automation and clinical application. The main advantage of 75Se-labeled methotrexate is its longer half-life, 121 days, as compared with 60 days for 125I

  13. Development of Solid Phase Radioimmunoassay for Measuring Methotrexate and 7-Hydroxy Methotrexate in Biological Fluids using Magnetizable Particles

    International Nuclear Information System (INIS)

    The objective of the present study was to prepare a solid phase magnetic particles radioimmunoassay (RIA) reagents. Development as well as optimization and validation of RIA system for the assessment of methotrexate (MTX) and its main metabolite 7-hydroxy methotrexate (7-OH MTX) in human serum were described. Preparation of 125 I-MTX and 125 I -7(OH) MTX were carried out using tyrosine methyl ester (TME) and chloramine-T (Ch-T) as oxidizing agent. HPLC column was used for purification of the prepared tracers. The production of polyclonal anti - bodies was carried out using six rabbits divided to two groups. The first one was immunized intraperitoneal through primary injection and three booster doses using MTX-BSA conjugate while the other group was immunized using 7(OH) MTX-BSA conjugate. Low density magnitizable cellulose iron oxide particles have been used to couple covalently to polyclonal antibody. The results obtained provide a low cost, simple, sensitive, specific, and accurate RIA system based on magnetizable solid phase separation.

  14. Spectrophotometric Determination of Trace Cyanide in Fruit Wines by the Catalytic Reaction of Ninhydrin Following Micro-Distillation

    Directory of Open Access Journals (Sweden)

    Saksit Chanthai

    2014-03-01

    Full Text Available Trace analysis of cyanide (CN based on the absorbance of the catalytic reaction of ninhydrin (NH in the presence of CN- was developed. This reaction was investigated consisting of 0.08 M NH, 0.4 M Na2CO3, 1% (v/v Tween 20 and 2.5 M NaOH in 5-mL final volume. The absorbance of the CN-NH complex was measured against the reagent blank at 598 nm. The calibration curve was widely linear over the range of 40-160 µg/L with r2 >0.99. The method recoveries of free cyanide, bound cyanide and total cyanide for wine samples were found in the range of 76.2-89.2%, 73.2-91.2% and 76.8-94.8%, respectively, at 250 µg/L CN- spiked level. Limit of detection was 6 µg/L. The reproducibility of the proposed method was less than 4.44%. This method was then applied for local Thai fruit wines. No trace amount of cyanide was detected, as if high recovery (88.4% of the micro-distillation was guaranteed. The obtained results were in agreement with those of the chloramine-T/barbituric acid-pyridine method with no statistically significant difference at 95% confidence level.

  15. Solid phase group specific absorbants in assays for glycoproteins

    International Nuclear Information System (INIS)

    The focus of this paper is on several technical advances in the assays for glycoprotein hormones and enzymes that have been achieved by use of the solid phase carbohydrate specific adsorbant concanavalin-A. Puriffication of glycoprotein radioligand after labelling by the chloramine-T method is readily accomplished using a small column of agarose bound concanavalin-A which separates glycoprotein radioligand from radioiodide and radiolabelled unadsorbed contaminants. After concanavalin-A column chromatography, radiolabelled glycoprotein hormone preparations exhibited improved binding to antibodies and tissue receptors. To increase the effective sensitivity of radioimmunoassays for glycoproteins, agarose bound concanavalin-A is used to extract and concentrate the glycoproteins from various biologic samples. For example, the effective sensitivity for the detection of human thyrotropin in serum was improved approximately 5 fold by using concanavalin-A concentrates of 1.5 ml of serum. Partial purification of the glycoprotein dopamine-β-hydroxylase from serum using agarose bound concanavalin-A resulted in separation of the serum factors that interfere with the measurement of enzyme activity. We conclude that in assays for glycoproteins, concanavalin-A is useful for purification of radioligand, for preparation of concentrates of glycoproteins from biologic samples, and for separation of glycoproteins from various interfering factors contained in biologic samples prior to radioligand or radioenzyme assay. (orig.)

  16. Solid-phase group-specific adsorbants in assays for glycoproteins

    International Nuclear Information System (INIS)

    The focus of the paper is on several technical advances in the assays for glycoprotein hormones and enzymes that have been achieved by the use of the solid-phase cabohydrate-specific adsorbant concanavalin-A. Purification of glycoprotein radioligand after labelling by the Chloramine-T method is readily accomplished using a small column of agarose-bound concanavalin-A which separates glycoprotein radioligand from radioiodide and radiolabelled unadsorbed contaminants. After concanavalin-A column chromatography, radiolabelled glycoprotein hormone preparations exhibited improved binding to antibodies and tissue receptors. To increase the effective sensitivity of radioimmunoassays for glycoproteins, agarose-bound concanavalin-A is used to extract and concentrate the glycoproteins from various biological samples. For example, the effective sensitivity for the detection of human thyrotropin in serum was improved approximately 5-fold by using concanavalin-A concentrates of 1.5ml of serum. Partial purification of the glycoprotein dopamine-β-hydroxylase from serum using agarose-bound concanavalin-A resulted in separation of the serum factors that interfere with the measurement of enzyme activity. We conclude that in assays for glycoproteins, concanavalin-A is useful for purification of radioligand, for preparation of concentrates of glycoproteins from biological samples and for separation of glycoproteins from various interfering factors contained in biological samples before radioligand or radioenzyme assay. (author)

  17. 125I Labelling of Protein Using Immobilized Enzyme

    International Nuclear Information System (INIS)

    For an effective solid-phase labelling of protein with 125I, studies on the immobilization of lactoperoxidase (LPO) on the inner wall of polystyrene tubes were carried out. Labelling of bovine serum albumin (BSA) and insulin was also practiced using the LPO immobilized tubes. The immobilized enzyme of about 2.5 μ g/tube was sufficient for small scale labelling since the results of radio-paper chromatography of the labelling mixture of insulin indicated that the yields were sufficiently high (80%) even in the reactions conducted at room temperature for 60 sec. The results of the Sephadex column chromatography indicated that the labelled products were not contaminated with LPO-125I, and the radiochemical purity of the products was more than 90%. In considering the general trend that the 125I labelled protein obtained by using LPO maintains its intactness better than those obtained by using chloramine-T, together with the tendency of yield enhancing with increase of reactants-concentration, the LPO immobilized tube method is estimated to be one of the simple methods of labelling. The product might be applicable without further purification.

  18. Improvement of labeling efficiency of glycoprotein-liposome conjugates with iodine-125 by using Bolton-Hunter Reagent and their distribution in mice

    Energy Technology Data Exchange (ETDEWEB)

    Kawakita, Yasunori; Kojima, Shuji [Science Univ. of Tokyo, Noda, Chiba (Japan). Research Institute for Biological Sciences; Yamazaki, Noboru

    2000-07-01

    We have developed a new type of glycoprotein-liposome conjugates and examined their potential utilities as drug-targeting carriers which exploit cellular functions of carbohydrate-binding proteins, i.e. animal lectins. An extremely low labeling efficiency, however, has been often a big problem in biodistribution study by using radiolabeled glycoprotein-liposome conjugates. In this study, improvement of the labeling efficiency was conducted by using Bolton-Hunter Reagent (BHR). First, tyrosyl groups were introduced into liposome membrane through amines of a constitutive phospholipid, dipalmitoylphosphatidlethanolamine (DPPE). Then, glycoprotein-tyrosyl group introduced liposomes were iodinated with {sup 125}I according to Chloramine-T methods. Labeling efficiency was markedly elevated in comparison with the BHR-untreated liposome conjugates. There was no significant changes in binding activity of BHR-treated glycoprotein-liposome conjugates with lectin. However, biodistribution of glycoprotein-tyrosyl group introduced liposomes in mice was significantly different from the mother conjugates. Thus, another suitable method for radioiodination of the glycoprotein-liposome conjugates should be developed. (author)

  19. Clinical scale preparation and evaluation of {sup 131}I-Rituximab for Non-Hodgkin's lymphoma

    Energy Technology Data Exchange (ETDEWEB)

    Kameswaran, Mythili; Vimalnath, K. Viswanathan; Rajeswari, Ardhi; Joshi, Prahlad Vasudeo; Samuel, Grace [Bhabha Atomic Research Centre, Mumbai (India). Radiopharmaceuticals Div.; Sarma, H.D. [Bhabha Atomic Research Centre, Mumbai (India). Radiation Biology and Health Sciences Div.

    2014-09-01

    Radioimmunotherapy (RIT) with anti CD20 MoAb conjugated to a β{sup -} emitting radioisotope like {sup 131}I or {sup 90}Y has the added advantage of delivering radiation not only to tumor cells that bind the antibody but also due to a crossfire effect, to neighboring tumor cells inaccessible to the antibody. In order to make available an indigenous radioimmunotherapeutic agent for Non Hodgkin's Lymphoma (NHL), radioiodinated Rituximab has been prepared and evaluated at a clinical scale. Radioiodination of Rituximab was performed by the conventional Chloramine T method using 7.4 GBq Na{sup 131}I in a lead shielded plant. Six batches of radioiodination were prepared and characterized by electrophoresis and HPLC to evaluate the reproducibility of the product. The product remained stable retaining the radiochemical purity > 95% upto 5 days after radioiodination. In vitro cell binding studies and biodistribution studies in normal Swiss mice have indicated the potential of this molecule as a radioimmunotherapeutic agent for NHL. (orig.)

  20. The transport of humic and fulvic acids through sand

    International Nuclear Information System (INIS)

    The objective of this work was to produce stable iodine-125 labelled humic and fulvic acids for use in field tests at the BGS borehole array at Drigg in Cumbria. The first part of the report describes iodine-125 labelling of humic and fulvic acids by oxidation of iodide-125 by chloramine-T. Although the apparent efficiency of labelling was high, some of the iodinated humic dissociated on standing and during passage through sand columns. The second part of the report describes the production of a more stable iodinated humic and fulvic acid. This was achieved by adding reducing agent to the reaction mixture after iodination of the humic material. The addition of reducing agent decreased the apparent labelling efficiency of the humic materials but increased the stability. The third part of the report describes the preparation of iodinated-fulvic acid for use in a field test. Gel column separation showed that 95% of the activity was bound to fulvic acid on the day of the field experiment. Samples of this material were passed through 55 cm long sand columns. I-125 breakthrough occurred simultaneously with tritium but a proportion remained on the column. This sorbed, or deposited, material then eluted very slowly from the column. (author)

  1. A radioimmunoassay of chicken growth hormone using growth hormone produced by recombinant DNA technology: validation and observations of plasma hormone variations in genetically fat and lean chickens

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) of chicken growth hormone (c-GH) has been developed using growth hormone produced by recombinant DNA technology. The best rabbit antiserum was used at 1/300,000 final dilution. Hormone labelling by iodine-125, achieved by chloramine T, allowed a specific activity of 3.7 MBq/μg. The equilibrium curves show that optimal conditions of incubation were reached at room temperature for 24h. This RIA used a second sheep antibody which precipitated the whole c-GH bound to the first antibody in the presence of polyethylene glycol solution (6%) at room temperature for 30 min. In our conditions, sensitivity was about 30 pg of c-GH per tube. Coefficient of variation was around 10%. No cross reaction was found with avian LH and prolactin. Thyrotrophin-releasing hormone (TRH) injection to young chickens induced 20-fold higher plasma c-GH concentrations. Simultaneous injection of somatostatin and TRH slightly reduced these concentrations. Hypoglycemia induced by insulin led to a drop of the plasma c-GH concentration. Conversely, refeeding or glucose load induced slight increases of the c-GH level. Genetically fat chickens tended to exhibit higher plasma c-GH concentrations than lean chickens

  2. Human C-peptide. Pt. 1

    International Nuclear Information System (INIS)

    Synthetic human C-peptide bearing a tyrosine group at its amino end is labelled with 125iodine using chloramin T or hydrogen peroxide and lactoperoxidase. The results of the two methods are compared. Antiserum to synthetic human C-peptide (without tyrosine), which was partially coupled to rabbit albumin, is raised in guinea pigs and goats. Goats show to be superior to guinea pips concerning antibody production. The so-called 'hook effect' phenomenon is observed when setting up the standard curves for the radioimmunoassay. Monotonically decreasing standard curves are obtained on dilution of antiserum with a high antibody titer which was produced by repeated immunization in goats. Free C-peptide and C-peptide bound to antiserum are separated using the anion exchange resin amberlite. Using this separation technique we excluded unspecific binding of labelled C-peptide to protein fractions in serum of diabetics. The sensitivity of our radioimmunoassay is approx. 0.3 ng C-peptide/ml serum. Intra- and interassay variability are below 10%. Human proinsulin is the only substance found to crossreact with the antiserum. (orig.)

  3. Measurement of insulin in human sera using a new RIA kit. 1

    International Nuclear Information System (INIS)

    A sensitive and versatile radioimmunoassay (RIA) for insulin was established using human insulin standard, a specific guinea pig anti-insulin antiserum and rabbit anti-guinea pig serum. Radioiodination was performed according to a modified chloramine T method. Tracer preparations were used for as long as 6 weeks after iodination. The standard curve ranges from 0.044 to 1.2 nmol/l. The intra-assay coefficient of variation (CV) was 3-5% and the inter-assay CV was 6-9% in the optimal range between 0.4 and 0.9 nmol/l. The average recovery of human insulin added to plasma or serum samples was 100.2 ± 2.0% (n = 38) and 100.1 ± 1.9% (n = 42), respectively. In addition to human insulin, procine, canine, rabbit and bovine insulin can also be determined but not rat or mouse insulin. The cross-reactivity of the antiserum with porcine proinsulin was found to be 40 % on the molar basis. The range of mean fasting plasma insulin concentrations in healthy subjects and under various pathological conditions were estimated. (author)

  4. Composition of cross-linked 125I-follitropin-receptor complexes

    International Nuclear Information System (INIS)

    Both of the alpha and beta subunits of intact human follitropin (FSH) were radioiodinated with 125I-sodium iodide and chloramine-T and could be resolved on sodium dodecyl sulfate-polyacrylamide gels. Radioiodinated FSH was affinity-cross-linked with a cleavable (nondisulfide) homobifunctional reagent to its membrane receptor on the porcine granulosa cell surface as well as to a Triton X-100-solubilized form of the receptor. Cross-linked samples revealed three additional bands of slower electrophoretic mobility, corresponding to 65, 83, and 117 kDa, in addition to the hormone bands. The hormone alpha beta dimer band corresponded to 43 kDa. Formation of the three bands requires the 125I-hormone to bind specifically to the receptor with subsequent cross-linking. Binding was prevented by an excess of the native hormone but not by other hormones. A monofunctional analog of the cross-linking reagent failed to produce the three bands. Reagent concentration-dependent cross-linking revealed that their formation was sequential; smaller complexes formed first and then larger ones. When gels of cross-linked complexes were treated to cleave covalent cross-links and then electrophoresed in a second dimension, 18-, 22-, and 34-kDa components were released, in addition to the alpha and beta subunits of the hormone

  5. Oxidative radioiodination damage to human lactoferrin

    International Nuclear Information System (INIS)

    Oxidative iodination of human lactoferrin (Lf) commonly performed by using the chloramine-T, the Iodogen or the lactoperoxidase method produces an unreliable tracer protein because of excessive and heterogeneous polymer formation. Before iodination a minor tetramer fraction may be demonstrable in iron-saturated Lf only. 125I-Lf polymers are mainly covalently linked, as suggested by the lack of substantial dissociation in SDS/polyacrylamide-gel electrophoresis. Damage to the 125I-Lf monomer may be another consequence of oxidative iodination. This is demonstrated in SDS/polyacrylamide-gel electrophoresis where 50% of the radioactivity of apparently normal monomer (Msub(r) 75000) is displaced to a lower-Msub(r) region (30000-67000) after reduction with dithiothreitol. Non-oxidative iodination by the Bolton-Hunter technique produces an antigenetically stable tracer that is not being subjected to polymerization and monomer degradation as judged by high performance gel chromatography and SDS/polyacrylamide-gel electrophoresis with and without dithiothreitol treatment. It is concluded that oxidation in itself leads to covalent non-disulphide cross-linking between human Lf molecules and, possibly, to intramolecular peptide-bond breaking becoming unmasked under reducing conditions. (author)

  6. Antiseptics and microcosm biofilm formation on titanium surfaces

    Directory of Open Access Journals (Sweden)

    Georgia VERARDI

    2016-01-01

    Full Text Available Abstract Oral rehabilitation with osseointegrated implants is a way to restore esthetics and masticatory function in edentulous patients, but bacterial colonization around the implants may lead to mucositis or peri-implantitis and consequent implant loss. Peri-implantitis is the main complication of oral rehabilitation with dental implants and, therefore, it is necessary to take into account the potential effects of antiseptics such as chlorhexidine (CHX, chloramine T (CHT, triclosan (TRI, and essential oils (EO on bacterial adhesion and on biofilm formation. To assess the action of these substances, we used the microcosm technique, in which the oral environment and periodontal conditions are simulated in vitro on titanium discs with different surface treatments (smooth surface - SS, acid-etched smooth surface - AESS, sand-blasted surface - SBS, and sand-blasted and acid-etched surface - SBAES. Roughness measurements yielded the following results: SS: 0.47 µm, AESS: 0.43 µm, SB: 0.79 µm, and SBAES: 0.72 µm. There was statistical difference only between SBS and AESS. There was no statistical difference among antiseptic treatments. However, EO and CHT showed lower bacterial counts compared with the saline solution treatment (control group. Thus, the current gold standard (CHX did not outperform CHT and EO, which were efficient in reducing the biofilm biomass compared with saline solution.

  7. Luminal oxidants selectively modulate electrogenic ion transport in rat colon

    Institute of Scientific and Technical Information of China (English)

    Julio M Mayol; Yolanda Adame-Navarrete; Pilar Alarma-Estrany; Elena Molina-Roldan; Fernando Huete-Toral; Jesus A Fernandez-Represa

    2006-01-01

    AIM: To investigate the effects of luminal exposure to H2O2 and two related thiol oxidizing agents on basal and stimulated chloride secretion in native colon using electrophysiological and pharmacological approaches.METHODS: Unstripped rat distal colon segments were mounted in Ussing chambers. Potential difference, cal culated resistance and short-circuit current across unstripped colon segments were monitored with a dual voltage/current clamp. Paracellular permeability was assessed by measuring the mucosa-to-serosa flux of a fluorescent probe (FITC).RESULTS: Luminal exposure to hydrogen peroxide transitorily stimulated chloride secretion without altering barrier function. This stimulatory effect could be blocked by basolateral atropine but not indomethacin. The cysteine and methionine oxidizing compounds, phenylarsine oxide and chloramine T respectively, mimicked the effect of H2O2, except for a drop in transcolonic resistance after 30 min. In contrast to the observed stimulatory effect on basal secretion, cAMP-stimulated electrogenic ion trans port was blunted by luminal H2O2. However, the Ca2+-activated response remained unchanged.CONCLUSION: H2O2 may be an important selective modulator of intestinal ion and water secretion in certain pathologic conditions such as inflammation or ischemiareperfusion by multiple mechanisms.

  8. Synthesis of 125 I - Salicyl Hydroxamic Acid for Urinary Bladder Imaging

    International Nuclear Information System (INIS)

    Salicylhydroxamic acid is a salicylate derivative. Radiolabeling of Salicyl hydroxamic acid ( SHA ) with iodine-125 may have considerable interest for imaging of urinary bladder. This study is aimed to optimize the radiolabeling yield of Salicyl hydroxamic with radio iodine (125-123) using chloramine - T (CAT) as an oxidizing agent with respect to factors that affect the reaction conditions such as SHA amount, CAT amount, reaction time and ph of the reaction mixture. In - vitro stability of the radiolabeled complex was checked and it was found to be stable for up to 24 h. 125 I-SHA was injected via intravenous administration routes into normal male Sprague – Dawley rats. Bio - distribution studies have revealed that 125I-SHA was excreted in urine with extent that it could give a clear image for urinary bladder especially if the bladder it tightly closed. The amount of 125 I - Salicyl hydroxamic excreted was increased in case of giving potassium bicarbonate to rat before injection of 125 I-SHA. The result of biodistribution study of 125 I - SHA in experimental animal suggest ed the possibility of using 123 I-SHA to image the urinary bladder

  9. Biochemical assays with radio-iodinated substrates

    International Nuclear Information System (INIS)

    A sensitive radiochemical method for the measurement of nanogram amounts of ammonia applicable to studies of enzymatic or other processes releasing or utilizing ammonia was developed. The method is based on a modified Berthelot reaction between ammonia and an aqueous alkaline solution of phenol and sodium hypochloride to yield indophenol. The radioactive tracer used is 125I-o-iodophenol, prepared by iodination of phenol with 125I by the chloramine-T method. A chloroform/carbon tetrachloride mixture is used to extract unreacted 125I-o-iodophenol from the reaction mixture; the 125I-labelled product remains in the aqueous phase and is measured by scintillation counting. Non-protein nitrogenous substances and certain other compounds interfere with the reaction, which cannot thus be used to measure ammonia directly in biological fluids. The glass Conway microdiffusion cell may, however, be utilized to isolate small amounts of ammonia from such fluids for measurement. The fluid is placed in the centre well of the cell and cold water in the outer well. Warm saturated potassium carbonate solution is then added to the centre well to expel the ammonia gas which is absorbed by the water. Technical details of the method are given

  10. Preparation of ( sup 123 I)- and ( sup 125 I)epidepride: a dopamine D-2 receptor antagonist radioligand

    Energy Technology Data Exchange (ETDEWEB)

    Clanton, J.A.; Schmidt, D.E.; Ansari, M.S.; Manning, R.G.; Kessler, R.M. (Vanderbilt Univ., Nashville, TN (United States). School of Medicine); Paulis, T. de (Vanderbilt Univ., Nashville, TN (United States). School of Medicine Vanderbilt Univ., Nashville, TN (United States). Dept. of Chemistry); Baldwin, R.M. (Yale Univ., West Haven, CT (United States). Veterans Administration Medical Center Research)

    1991-07-01

    (S)-(-)-N-((1-ethyl-2-pyrrolidinyl)methyl)-5-({sup 123}I) iodo-2,3-dimethoxybenzamide (TDP 517) (proposed generic name, ({sup 123}I)epidepride) is the iodine-123 substituted analogue of isoremoxipride (FLB 457), both of which are very potent dopamine D-2 antagonists (epidepride K{sub D} 0.024 nM). ({sup 123}I) Epidepride was radioiodinated in 60-70% radiochemical yields in 35 min from the corresponding 5-(tributyltin) derivative using Na{sup 123}I with a specific radioactivity of 3000 Ci/mmol, and oxidized in situ with chloramine-T. The aryltin precursor was prepared from non-labelled epidepride by palladium-catalyzed stannylation using bis(tri-n-butyltin) in triethylamine. Alternatively, using no carrier-added Na{sup 125}I as the radioisotope, ({sup 125}I) epidepride at 2000 Ci/mmol specific radioactivity was prepared in 86% radiochemical yield and 99% radiochemical purity after purification by reverse phase HPLC in ethanolic phosphate buffer. (author).

  11. Preparation of 125I labelled nerve growth factor and study of its pharmacokinetics

    International Nuclear Information System (INIS)

    125I-Nerve growth factor (125I-NGF) with more than 95% of radiochemical purity is prepared by chloramine-T method. After I.V. and I.M. injection of 125I-NGF, pharmacokinetics of NGF in mice is determined by SDS-PAGE method. The results show that concentration-time curves after I.V. injection are fitted to a 2-compartment model are those after I.M. injection are fitted to a 1-compartment model. After I.V. injection of 25 μg/kg, the elimination half life (t1/2(β)) is 3.65 h. After I.M. injection of 75, 25, 10, 3.3 μg/kg, t1/2(β) is 1.79, 2.25, 2.30, 3.24 h, respectively. The mean plasma clearance (CLs) is 0.37 L/(h·kg), the appearance volume of distribution (Vd) is 1.18 L/kg and the mean residence time (tr-bar) in mice is 2.78 h. The pharmacokinetics of 125I-NGF in mice is useful for clinical trial

  12. Iodine-123 labeled derivatives of methylphenidate: potential SPECT radiopharmaceuticals for brain dopamine transporters

    Energy Technology Data Exchange (ETDEWEB)

    Pan, D.; Gatley, S.J.; Chen, R.; Ding, Y.-S. [Brookhaven National Lab., Upton, NY (United States)

    1996-06-01

    Since dl-threo-[{sup 11}C]methylphenidate (Ritalin) and especially the more active enantiomer, d-threo-[{sup 11}C]methylphenidate, have favorable properties for PET studies, we prepared two radioiodinated analogs of methylphenidate, p-[{sup 123}I]iodomethylphenidate and m-[{sup 123}I]iodo-p-hydroxymethylphenidate with a view to evaluating them as potential SPECT tracers. To prepare p-[{sup 123}I]iodomethylphenidate, the p-tributyltin derivative was prepared from the previously reported p-bromomethylphenidate and reacted under acidic conditions with I-123 iodide plus chloramine-T at room temperature for 90 seconds. The predomimant radioactive product was obtained in 85% radiochemical yield and > 10 Ci/{mu}mol specific radioactivity after HPLC purification. It had the same HPLC retention time as a spectroscopically characterized non-radioactive p-iodomethylphenidate standard prepared via nitration of methylphenidate and diazotization, after protection of the secondary amino group by benzoylation. A second radioiodinated methylphenidate derivative, m-[{sup 123}I]iodop-hydroxymethylphenidate was prepared in 80% radiochemical yield by direct iodination of the known p-hydroxymethylphenidate. In this case the non-radioactive standard was prepared by iodination of p-hydroxyritalinic acid using I{sub 2} and iodic acid, followed by esterification. (author).

  13. Radioiodination of chicken erythrocyte histones H4 and H5 in chromatin.

    Science.gov (United States)

    Griffiths, G R; Huang, P C

    1979-08-25

    The conformational state of histones in isolated chicken erythrocyte chromatin was studied using procedures developed for probing surface proteins on membranes. Under controlled conditions, only exposed tyrosyl residues react with iodide radicals, generated either by the oxidant, chloramine-T (paratoluenesulfonyl chloramide), or the enzyme lactoperoxidase, giving monoidotyrosine. Using 125-iodine, this study compared the reactive tyrosines in free and bound histones H4, and H5. The relative extent of iodination of these histones within (H4) and outside (H5) of the nucleosomes was measured after extraction and gel electrophoresis. Each of the histones was further analyzed for the extent of specific tyrosine iodination by separating the tryptic peptides by high voltage electrophoresis. The identity of the labeled peptide was determined by dansylation of the amino acids present in each hydrolyzed peptide. The results show that there is a difference in the conformational arrangement of these histones on chromatin and in the free forms, since in chromatin not all tyrosine residues are as accessible for iodination as in the denatured state. Residue 53 of histone H5 for instance is more reactive than residues 28 and 58, indicating that the segments containing the latter residues are involved in either protein-DNA or protein-protein interactions. In histone H4, preferential labeling of 2 of the 4 tyrosines present was also observed.

  14. Plasma half-life and organ uptake ratio of radiolabeled glandular kallikrein in control and nephrectomized rats

    Energy Technology Data Exchange (ETDEWEB)

    Nishimura, K.; Iwata, T.; Kokubu, T.

    1986-01-01

    The purified rat urinary kallikrein was radiolabeled by lactoperoxidase method and by chloramine T method. Plasma half-life of radiolabeled kallikrein was 5.06 +/- 0.59 (n = 5) min in control rats and 5.24 +/- 0.42 (n = 5) min in nephrectomized rats. There was no difference between two groups. From autoradiogram, main metabolic organs of radiolabeled kallikrein were liver, kidney and spleen. Total uptake of radiolabeled kallikrein in ech organ was the highest in liver (73.2%). The uptake per g tissue of radiolabeled kallikrein in each organ was high in liver (33.0%), kidney (31.4%) and spleen (21.1%). These results suggest that the active kallikrein is metabolized mainly in the liver, and kidney is not so an important organ to metabolize or to eliminate the active kallikrein in plasma. In order to clarify the mode of existence of active kallikrein in plasma, the following experiment was done by using disc gel electrophoresis. Radioactive profile of radiolabeled kallikrein showed one peak (Rf = 1.0), but radiolabeled kallikrein mixed with rat plasma showed two peaks, that is small peak (Rf = 1.0), and main peak (RF = 0.5). The most of radiolabeled kallikrein was bound to plasma protein and only five per cent was in free form. Furthermore, the binding of radiolabeled kallikrein to plasma protein was interfered by the addition of active kallikrein. These results suggest the possibility of existence of kallikrein binding protein in plasma.

  15. Oxidative radioiodination damage to human lactoferrin

    Energy Technology Data Exchange (ETDEWEB)

    Rosenmund, A.; Kuyas, C.; Haeberli, A.

    1986-11-15

    Oxidative iodination of human lactoferrin (Lf) commonly performed by using the chloramine-T, the Iodogen or the lactoperoxidase method produces an unreliable tracer protein because of excessive and heterogeneous polymer formation. Before iodination a minor tetramer fraction may be demonstrable in iron-saturated Lf only. /sup 125/I-Lf polymers are mainly covalently linked, as suggested by the lack of substantial dissociation in SDS/polyacrylamide-gel electrophoresis. Damage to the /sup 125/I-Lf monomer may be another consequence of oxidative iodination. This is demonstrated in SDS/polyacrylamide-gel electrophoresis where 50% of the radioactivity of apparently normal monomer (Msub(r) 75000) is displaced to a lower-Msub(r) region (30000-67000) after reduction with dithiothreitol. Non-oxidative iodination by the Bolton-Hunter technique produces an antigenetically stable tracer that is not being subjected to polymerization and monomer degradation as judged by high performance gel chromatography and SDS/polyacrylamide-gel electrophoresis with and without dithiothreitol treatment. It is concluded that oxidation in itself leads to covalent non-disulphide cross-linking between human Lf molecules and, possibly, to intramolecular peptide-bond breaking becoming unmasked under reducing conditions.

  16. Selective mono-radioiodination and characterization of a cell-penetrating peptide. L-Tyr-maurocalcine

    Energy Technology Data Exchange (ETDEWEB)

    Ahmadi, Mitra; Bacot, Sandrine; Perret, Pascale; Riou, Laurent; Ghezzi, Catherine [Universite Joseph Fourier, Grenoble (France); INSERM U1039, Grenoble (France). Radiopharmaceutiques Biocliniques; Poillot, Cathy; Cestele, Sandrine [INSERM U836, Grenoble (France). Grenoble Inst. of Neuroscience; Universite Joseph Fourier, Grenoble (France); Desruet, Marie-Dominique [INSERM U1039, Grenoble (France). Radiopharmaceutiques Biocliniques; Couvet, Morgane; Bourgoin, Sandrine; Seve, Michel [CRI-INSERM U823, Grenoble (France). Inst. of Albert Bonniot; Universite Joseph Fourier, Grenoble (France); Waard, Michel de [INSERM U836, Grenoble (France). Grenoble Inst. of Neuroscience; Universite Joseph Fourier, Grenoble (France); Smartox Biotechnologies, Grenoble (France)

    2014-07-01

    Mono-and poly-iodinated peptides form frequently during radioiodination procedures. However, the formation of a single species in its mono-iodinated form is essential for quantitative studies such as determination of tissue concentration or image quantification. Therefore, the aim of the present study was to define the optimal experimental conditions in order to exclusively obtain the mono-iodinated form of L-maurocalcine (L-MCa). L-MCa is an animal venom toxin which was shown to act as a cell-penetrating peptide. In order to apply the current direct radioiodination technique using oxidative agents including chloramine T, Iodo-Gen {sup registered} or lactoperoxidase, an analogue of this peptide containing a tyrosine residue (Tyr-L-MCa) was synthesized and was shown to fold/oxidize properly. The enzymatic approach using lactoperoxidase/H{sub 2}O{sub 2} was found to be the best method for radioiodination of Tyr-L-MCa. MALDI-TOF mass spectrometry analyses were then used for identification of the chromatographic eluting components of the reaction mixtures. We observed that the production of different radioiodinated species depended upon the reaction conditions. Our results successfully described the experimental conditions of peptide radioiodination allowing the exclusive production of the mono-iodinated form with high radiochemical purity and without the need for a purification step. Mono-radioiodination of L-Tyr-MCa will be crucial for future quantitative studies, investigating the mechanism of cell penetration and in vivo biodistribution.

  17. Preparation and properties of /sup 3/H-phe/sup B1/ insulin

    Energy Technology Data Exchange (ETDEWEB)

    Grant, K.I.

    1986-01-01

    Insulin iodinated by both the lactoperoxidase and chloramine-T methods was found in accordance with many published reports to bind to plasma membrane. It was found, however not to obey isotope dilution laws in its displacement from plasma membrane by natural insulin. It was shown to be heterogeneous on reverse-phase HPLC, to contain diiodotyrosine and to have a reduced biological activity in an in vivo assay. Thus an insulin labelled semi-synthetically with /sup 3/H-phenylalaine at position B1 was prepared. A method already published was attempted, but without success. A different scheme of synthesis was developed using 2-t-butoxycarbonyloxyimino-2-phenylacetonitrile as the t-butoxycarbonylating agent and HPLC as the mode of separation of insulin derivatives. Using L-(2,3,4,5,6-/sup 3/H) phenylalaine a labelled insulin of specific radioactivity of 60 Ci/mmole was made. Characterization by the techniques of HPLC, acid-urea polyacrylamide gel electrophoresis, dansylation, Edman degradation and gel exclusion chromatography showed the labelled insulin to be indistinguishable from the natural hormone, and to contain a single covalently bound /sup 3/H-phe residue.

  18. Biologically active monoiodinated alpha-MSH derivatives for receptor binding studies using human melanoma cells

    International Nuclear Information System (INIS)

    Three different monoiodinated radioligands of alpha-MSH (alpha-melanocyte-stimulating hormone) were compared in a binding assay with human D10 melanoma cells: [Tyr(125I)2]-alpha-MSH, [Tyr(125I)2,NIe4]-alpha-MSH, and [Tyr(125I)2,NIe4,D-Phe7]-alpha-MSH. They were prepared either by the classical chloramine T method or by the Enzymobead method. A simple and rapid purification scheme was developed consisting of a primary separation on reversed-phase C18 silica cartridges immediately after the iodination, followed by HPLC purification before each binding experiment. Biological testing of the three radioligands showed that they all retained high melanotropic activity in the B16 melanin assay and the Anolis melanophore assay. However, in human D10 melanoma cells, [Tyr(125I)2,NIe4]-alpha-MSH led to a high degree of non-specific binding to the cells which could not be displaced by excess alpha-MSH and only partially by [NIe4]-alpha-MSH. The [Tyr(125I)2,NIe4,D-Phe7]-alpha-MSH tracer gave similar results but with a much lower proportion of non-specific binding. On the other hand, [Tyr(125I)2]-alpha-MSH proved to be an excellent radioligand whose non-specific binding to the D10 cells was not higher than 20% of the total binding

  19. Short communication: rapid preparation of preventive and therapeutic whole-killed retroviral vaccines using the microbicide taurine chloramine.

    Science.gov (United States)

    Dudani, A K; Martyres, A; Fliss, H

    2008-04-01

    A current urgent priority is to develop microbicides and vaccines to combat retroviruses like human immunodeficiency virus (HIV). We show that the cysteine-selective natural compound, taurine chloramine (T-NCl), can be effective in this task. A number of proteins in all retroviruses contain highly conserved cysteine-rich regions that are essential for infection and replication. Our data show that by targeting these essential cysteine residues, T-NCl (2 or 5 mM) acts as a highly effective and safe microbicide that fully blocks the infectivity of high HIV-1 titers (10(6) TCID(50) units/ml) but is not injurious to eukaryotic cells. We also demonstrate that T-NCl can be used to prepare a highly effective whole-killed vaccine against murine AIDS (MAIDS) that shows both preventive and therapeutic efficacy. The vaccine consists of a T-NCl-inactivated retrovirus suspension in host cell lysate. The novelty of our approach lies in the ease and speed of vaccine preparation and its avoidance of harsh inactivation or purification steps that can alter native viral conformation. Our approach is therefore likely to overcome a number of intractable obstacles to the preparation of an effective whole-killed HIV vaccine, such as surviving infective viral particles, rapid viral mutation rates, numerous viral strains, and harsh purification steps. Our approach may also permit the rapid preparation of autologous, or custom-made, vaccines for individual patients.

  20. Selective mono-radioiodination and characterization of a cell-penetrating peptide. L-Tyr-maurocalcine

    International Nuclear Information System (INIS)

    Mono-and poly-iodinated peptides form frequently during radioiodination procedures. However, the formation of a single species in its mono-iodinated form is essential for quantitative studies such as determination of tissue concentration or image quantification. Therefore, the aim of the present study was to define the optimal experimental conditions in order to exclusively obtain the mono-iodinated form of L-maurocalcine (L-MCa). L-MCa is an animal venom toxin which was shown to act as a cell-penetrating peptide. In order to apply the current direct radioiodination technique using oxidative agents including chloramine T, Iodo-Gen registered or lactoperoxidase, an analogue of this peptide containing a tyrosine residue (Tyr-L-MCa) was synthesized and was shown to fold/oxidize properly. The enzymatic approach using lactoperoxidase/H2O2 was found to be the best method for radioiodination of Tyr-L-MCa. MALDI-TOF mass spectrometry analyses were then used for identification of the chromatographic eluting components of the reaction mixtures. We observed that the production of different radioiodinated species depended upon the reaction conditions. Our results successfully described the experimental conditions of peptide radioiodination allowing the exclusive production of the mono-iodinated form with high radiochemical purity and without the need for a purification step. Mono-radioiodination of L-Tyr-MCa will be crucial for future quantitative studies, investigating the mechanism of cell penetration and in vivo biodistribution.

  1. Studies into the radiobromination and -iodination of aromatic compounds with n-halogen compounds without addition of carriers

    International Nuclear Information System (INIS)

    There is a special need for radiohalogenated compounds for diagnostic nuclear medicine, in particular for no-carrier-added (nca) short lived cyclotron products. In this study the applicability of chloramine-T (CAT) and dichloramine-T (DCT) has been demonstrated for nca-radiohalogenation with bromine-75, 77 and iodine-123 in aqueous and organic solvents. Radio gaschromatography and -HPLC are used for product analysis. In aqueous solution the optimum reaction conditions with respect to pH, concentration of CAT, reaction time and added bromide-carrier are determined using tyrosine as a model substrate. The suitability of the CAT method for radiobrominations under convenient and mild conditions is demonstrated for some biomolecules such as amino acids and nucleobases as well as for peptides and the proteins urokinase and HSA. Dichloramine-T is found to be a new efficient reagent for radiobromination and -iodination of aromatic compounds in various organic solvents such as acetic acid, dichloromethane or carbontetrachloride. A high para-selectivity is observed and the radiochemical yields (80%) are as high as in aqueous solution. A comparison of this reagent with different in-situ halogenation agents show that DCT is superior with respect to reaction time, concentration of the reagent and thus oxidative side reactions, and ease of handling. (orig./RB)

  2. Variation of hydroxyproline content after local injection hyaluronic acid in excessive dermal scar of rabbit ears%透明质酸注射兔耳瘢痕后羟脯氨酸含量变化

    Institute of Scientific and Technical Information of China (English)

    张阳; 柳大烈; 李希军; 李玉霞; 张选奋; 安书杰

    2002-01-01

    AIM To observe the scar formation by using hyaluronic acid (HA) and hyaluronidase (HAse) in excessive dermal scar of rabbit ears. METHODS Upon the foundation of hypertrophic scar model in rabbit ears, we tested the content of hydroxyproline (HPr) using chloramineT method by injecting HA and HAse in excessive dermal scar of rabbit ears with the establishment of saline and blank control at different time. RESULTS ① HPr contents of the HA, saline and blank groups were gradullay increased (P0.05); ③ Compared with saline group, HPr contents of the HA group were higher (P0.05);③HA组的HPr含量明显高于盐水组(P<0.05);④HAse组的HPr含量明显低于盐水组(P<0.01);⑤HA组的HPr含量明显高于HAse组(P<0.01).结论瘢痕组织中注射HA后胶原合成增加, 提示瘢痕组织中, HA含量的持续增高可导致瘢痕的异常增生.

  3. Studies on the mode of action of calciferol. XIII. Development of a radioimmunoassay for vitamin D-dependent chick intestinal calcium-binding protein and tissue distribution

    International Nuclear Information System (INIS)

    A RIA for chick intestinal calcium-binding protein (CaBP) has been developed with a sensitivity of 1 ng. The antiserum was generated in rabbits injected with highly purified vitamin D-dependent chick intestinal CaBP. The assay employs the double antibody technique, and 125I-labeled CaBP was prepared using chloramine T. Low molecular weight peptide hormones and normal rabbit, rat, and human serum proteins show no cross-reactivity in the assay. Measurements of chick intestinal and kidney CaBP by RIA showed a good correlation with measurements of CaBP by the radial immunodiffusion method. The assay is reproducible (interassay variability, 16.3%) and precise (intraassay variability, 4.0%). The concentration of immunoreactive CaBP (iCaBP) in chick serum (2.7 ng/ml serum) can now be measured as early as 8 h after the administration of 6.5 nmol 1,25-dihydroxyvitamin D3; a maximum of 11 ng/ml is reached at 20 h. The level of CaBP in chick serum was found to be dependent on the dose of vitamin D3 or 1,25-dihydroxyvitamin D3 administered to the animal

  4. Studies on the mode of action of calciferol. XIII. Development of a radioimmunoassay for vitamin D-dependent chick intestinal calcium-binding protein and tissue distribution

    Energy Technology Data Exchange (ETDEWEB)

    Christakos, S.; Friedlander, E.J.; Frandsen, B.R.; Norman, A.W.

    1979-05-01

    A RIA for chick intestinal calcium-binding protein (CaBP) has been developed with a sensitivity of 1 ng. The antiserum was generated in rabbits injected with highly purified vitamin D-dependent chick intestinal CaBP. The assay employs the double antibody technique, and /sup 125/I-labeled CaBP was prepared using chloramine T. Low molecular weight peptide hormones and normal rabbit, rat, and human serum proteins show no cross-reactivity in the assay. Measurements of chick intestinal and kidney CaBP by RIA showed a good correlation with measurements of CaBP by the radial immunodiffusion method. The assay is reproducible (interassay variability, 16.3%) and precise (intraassay variability, 4.0%). The concentration of immunoreactive CaBP (iCaBP) in chick serum (2.7 ng/ml serum) can now be measured as early as 8 h after the administration of 6.5 nmol 1,25-dihydroxyvitamin D/sub 3/; a maximum of 11 ng/ml is reached at 20 h. The level of CaBP in chick serum was found to be dependent on the dose of vitamin D/sub 3/ or 1,25-dihydroxyvitamin D/sub 3/ administered to the animal.

  5. Dynamic behavior of histone H1 microinjected into HeLa cells

    International Nuclear Information System (INIS)

    Histone H1 was purified from bovine thymus and radiolabeled with tritium by reductive methylation or with 125I using chloramine-T. Red blood cell-mediated microinjection was then used to introduce the labeled H1 molecules into HeLa cells synchronized in S phase. The injected H1 molecules rapidly entered HeLa nuclei, and a number of tests indicate that their association with chromatin was equivalent to that of endogenous histone H1. The injected molecules copurified with HeLa cell nucleosomes, exhibited a half-life of ∼100h, and were hyperphosphorylated at mitosis. When injected HeLa cells were fused with mouse 3T3 fibroblasts < 10% of the labeled H1 molecules migrated to mouse nuclei during the next 48 h. Despite their slow rate of migration between nuclei, the injected H1 molecules were evenly distributed on mouse and human genomes soon after mitosis of HeLa-3T3 heterokaryons. These results suggest that although most histone H1 molecules are stably associated with interphase chromatin, they undergo extensive redistribution after mitosis

  6. The study of labeling with Iodine-131 of monoclonal antibody anti-CD20 used for the treatment of non-Hodgkin lymphoma

    International Nuclear Information System (INIS)

    Lymphomas are malignancies of the lymphatic system, described by Thomas Hodgkin in 1932. Traditionally, lymphomas are classified in two basic groups: Hodgkin disease and non-Hodgkin lymphoma (NHL). Patients with NHL were earlier treated with radiotherapy alone or in combination with immunotherapy using monoclonal antibody anti-CD20 (ex., Rituximab-Mabthera, Roche). However, Radioimmunotherapy is a new modality of treatment for patients with NHL, in which cytotoxic radiation from therapeutic radioisotopes is delivered to tumors through monoclonal antibodies. This study focused on labeling conditions of monoclonal antibody anti-CD20 (Rituximab-Mabthera, Roche) with iodine-131, by direct radioiodination method using Chloramine-T as oxidizing agent. Labeling parameters investigated were: Radiochemical purity (RP), method of purification, incubation time, antibody mass, oxidative agent mass, stability in vitro, stability in vivo, immunoreactivity and biological distribution performed in normal Swiss mouse. Product of high radiochemical purity was obtained with no notable difference between the methods applied. No clear evidence of direct influence of incubation time on radiochemical purity of the labeled antibody was observed. Whereas, a clear evidence of direct influence of activity on radiochemical purity of the labeled antibody was observed when antibody mass was varied. After purification, the labeled product presented radiochemical purity of approximately 100 %. Product of superior radiochemical yield was observed when standard condition of labeling was used. The labeled product presented variation in radiochemical purity using five different stabilizer conditions. The condition in which gentisic acid was combined with freeze appears more suitable and capable of minimizing autoradiolysis of the antibody labeled with high therapeutic activity of iodine-131. The labeled product presented low immunoreactivity when compared to the literature. Biological distribution in

  7. The study of labeling with iodine-131 of monoclonal antibody anti-CD20 used for the treatment of non-Hodgkin lymphoma

    International Nuclear Information System (INIS)

    Lymphomas are malignancies of the lymphatic system, described by Thomas Hodgkin in 1932. Traditionally, lymphomas are classified in two basic groups: Hodgkin disease and non-Hodgkin lymphoma (NHL). Patients with NHL were earlier treated with radiotherapy alone or in combination with immunotherapy using monoclonal antibody anti-CD20 (ex., Rituximab-Mabthera, Roche). However, Radioimmunotherapy is a new modality of treatment for patients with NHL, in which cytotoxic radiation from therapeutic radioisotopes is delivered to tumors through monoclonal antibodies. This study focused on labeling conditions of monoclonal anti-CD20 (ex., Rituximab-Mabthera, Roche) with iodine-131, by direct radioiodination method using Chloramine-T as oxidizing agent. Labeling parameters investigated were: Radiochemical purity (RP), method of purification, incubation time, antibody mass, oxidative agent mass, stability in vitro, immunoreactivity and biological distribution performed in normal Swiss mouse. Product of high radiochemical purity was obtained with no notable difference between the methods applied. No clear evidence of direct influence of incubation time on radiochemical purity of the labeled antibody was observed. Whereas, a clear evidence of direct influence of activity on radiochemical purity of the labeled antibody was varied. After purification the labeled product presented radiochemical purity of approximately 100 %. Product of superior radiochemical yield was observed when standard condition of labeling was used. The labeled product presented variation in radiochemical purity using five different stabilizer conditions. The condition in which gentisic acid combined with freeze appears more suitable and capable of minimizing autoradiolysis of the antibody labeled with freeze appears more suitable and capable of minimizing autoradiolysis of the antibody labeled with high therapeutic activity of iodine-131. The labeled product presented low immunoreactivity when compared to the

  8. The study of labeling with Iodine-131 of monoclonal antibody anti-CD20 used for the treatment of non-Hodgkin lymphoma; Estudo de marcacao com Iodo-131 de anticorpo monoclonal anti-CD20 na terapia de linfoma nao-Hodgkin

    Energy Technology Data Exchange (ETDEWEB)

    Akanji, Akinkunmi Ganiyu

    2006-07-01

    Lymphomas are malignancies of the lymphatic system, described by Thomas Hodgkin in 1932. Traditionally, lymphomas are classified in two basic groups: Hodgkin disease and non-Hodgkin lymphoma (NHL). Patients with NHL were earlier treated with radiotherapy alone or in combination with immunotherapy using monoclonal antibody anti-CD20 (ex., Rituximab-Mabthera, Roche). However, Radioimmunotherapy is a new modality of treatment for patients with NHL, in which cytotoxic radiation from therapeutic radioisotopes is delivered to tumors through monoclonal antibodies. This study focused on labeling conditions of monoclonal antibody anti-CD20 (Rituximab-Mabthera, Roche) with iodine-131, by direct radioiodination method using Chloramine-T as oxidizing agent. Labeling parameters investigated were: Radiochemical purity (RP), method of purification, incubation time, antibody mass, oxidative agent mass, stability in vitro, stability in vivo, immunoreactivity and biological distribution performed in normal Swiss mouse. Product of high radiochemical purity was obtained with no notable difference between the methods applied. No clear evidence of direct influence of incubation time on radiochemical purity of the labeled antibody was observed. Whereas, a clear evidence of direct influence of activity on radiochemical purity of the labeled antibody was observed when antibody mass was varied. After purification, the labeled product presented radiochemical purity of approximately 100 %. Product of superior radiochemical yield was observed when standard condition of labeling was used. The labeled product presented variation in radiochemical purity using five different stabilizer conditions. The condition in which gentisic acid was combined with freeze appears more suitable and capable of minimizing autoradiolysis of the antibody labeled with high therapeutic activity of iodine-131. The labeled product presented low immunoreactivity when compared to the literature. Biological distribution in

  9. Two Fatal Intoxications with Cyanohydrins.

    Science.gov (United States)

    Zheng, Shuiqing; Yuan, Xiaoliang; Wang, Wei; Liang, Chen; Cao, Fangqi; Zhang, Runsheng

    2016-06-01

    Cyanohydrins, also be called cyanoalcohols, are important industrial precursors to carboxylic acids and some amino acids. Acetone cyanohydrin (ACH) and formaldehyde cyanohydrin (glycolonitrile, FCH), which are the typical examples of cyanohydrins, are classified as extremely hazardous substances. As the cyanohydrins can readily decompose, and it is hard to find cyanohydrins in gastric contents and heart blood, the determination study in biological samples can be divided into two parts: the first is the determination of HCN by using a Prussian blue reaction and the HS-GC-MSD after derivatization by chloramine-T. The second is the determination of acetone or formaldehyde. In this part, headspace gas chromatography with flame ionization detector (HS-GC-FID) and solid phase microextraction (SPME)-gas chromatography with mass spectrometric detectors (GC-MSD) had been used. In this report, we reported two fatal intoxication cases of ACH and FCH; one person was killed by his wife by poisoning his food and the other was suicide by poison. Two real cases of ACH and FCH in human blood and gastric contents have been analyzed by using the above-mentioned method. The Prussian blue reaction was positive in the two cases. The peaks of acetone with retention times of 0.998 min appear in specimens of the deceased are consistent with the retention times of pure acetone. The peaks of formaldehyde with a retention time of 1.658 min appear in heart blood of the deceased, and the retention time of formaldehyde of the liquid is 1.674 min, which are consistent with the retention times of pure formaldehyde (1.673 min). PMID:27026650

  10. N-Chlorotaurine and ammonium chloride: an antiseptic preparation with strong bactericidal activity.

    Science.gov (United States)

    Gottardi, Waldemar; Arnitz, Roland; Nagl, Markus

    2007-04-20

    The bactericidal activity of the endogenous antiseptic N-chlorotaurine (NCT) is significantly enhanced in the presence of ammonium chloride which induces the formation of monochloramine (NH(2)Cl) whose strong bactericidal activity is well known. In this study the properties of NCT plus ammonium chloride have been investigated. The reaction of active chlorine compounds like chloramine-T (N-chlorotoluene-sulfonamide sodium), chloroisocyanuric acid derivatives, hypochlorites (NaOCl, CaOCl(2)) with ammonium chloride did not stop at the stage of monochloramine, and the pungent smelling by-products di- and trichloramine, NHCl(2) and NCl(3), were also formed. This was not the case with NCT where only monochloramine was generated. The equilibrium constant of the reaction of NCT with ammonium was found to be [Formula: see text] , which allows to estimate the equilibrium concentration of monochloramine in aqueous solutions of NCT and ammonium chloride. At concentrations each ranging between 0.01% and 1.0% it comes to [NH(2)Cl]=3.5-254 ppm. As an unexpected result the monochloramine containing formulation turned out to be most stable in plain water without buffer additives. Quantitative killing assays revealed complete inactivation of 10(6) to 10(7)CFU/mL of seven bacterial strains by 0.1% NCT plus 0.1% ammonium chloride within 5 min, while with plain 0.1% NCT an incubation time of 2-4h was needed to achieve the same effect. The highly significant increase of bactericidal activity (200-300-fold) could be assigned to the presence of monochloramine which could be isolated by vacuum distillation. Aqueous solutions of NCT and ammonium chloride provide a highly effective and well tolerable antiseptic preparation appropriate to a treatment cycle of at least 1 month if stored in the refrigerator.

  11. New bisphosphonate labeled with Iodine-131 for the palliative therapy for bone metastases pain

    International Nuclear Information System (INIS)

    The aim of this work was to obtain new bisphosphonate marked with 131I suitable for palliative treatment of bone metastases pain characteristics. Materials and Methods: It started with aromatic amino acids and the synthesis consisted of three stages: 1) Protection of amino groups by acetylation; 2) phosphonation protected amino acids with a mixture of phosphorous acid and phosphorus pentachloride; 3) Lack of protection of the amino groups by basic hydrolysis. The compounds obtained were characterized by IR, 1H NMR, RMN13-C mass. Los spectrometry bisphosphonic acids obtained were labeled with 131I using chloramine T and iodogen as oxidants. Stability of labeled compounds in aqueous solution was studied serum. 3 mg of 2-amino-3- (4-hydroxyphenyl) -1-hydroxypropyl-1,1-bisphosphonic acid labeled of 131I were administered to male wistar rats (170-190 g) through a lateral tail vein. The scintigraphic study was conducted at 2, 6 and 12 hours. Results: The yield of the reactions of the amino group protection four compounds ranged from 75 to 80%, while the phosphonation was between 50 and 60%. The radiochemical purity of 2-amino-3- (4-hydroxyphenyl) -1-hydroxypropyl-1,1- bisphosphonic acid labeled with 131I was (91.5 ± 1.4)% and its stability was satisfactory for 72h. Scintigraphic images suggest excretion by the kidneys of the compound and from 12 h post-administration begin to visualize bone structures of the animal, suggesting that the compound exhibits affinity for these tissues. Conclusions: A novel synthesis method with modifications that yielded the sodium salts of bisphosphonic acids starting from the respective aromatic amino acids was developed. 2-amino-3- (4-hydroxyphenyl) -1-hydroxypropyl-1,1-bisphosphonic acid 131I labeled was stable up to 72h and showed affinity for bone tissue. (author)

  12. Preparation of monoclonal antibodies against cardiac myosin and some radiolabelling studies

    International Nuclear Information System (INIS)

    Monoclonal antibodies were raised against myosin, a specific indicator of myocardial infarction and labelled with 125I and 99mTc. Human cardiac myosin was isolated from normal human heart and was used for raising the monoclonal antibodies by the hybridoma technique. Antibody producing clones were identified by ELISA and cloning was done by the limiting dilution technique. Of the 13 clones obtained, 4 were deemed suitable for further studies. The antibodies were grown in ascites, purified, isotyped and their cross reactions with other forms of myosin were estimated. All the clones showed negligible cross reaction with rabbit myosin, but reacted to different extents with bovine skeletal myosin. The most avid antibody Mab-4G4 was chosen for further labelling studies. Mab-4G4 was labelled with 125I using different oxidising agents such as iodogen, chloramine-T and lactoperoxidase. Purified radioiodinated antibody with radiochemical purity >95% could be obtained by gel filtration. Immunoreactivity was retained as tested by binding to myosin immobilised on a solid support. Mab-4G4 was also labelled with 99mTc using stannous tartrate as the reducing agent. Radiolabelling yield was ∼60%, the purity was >95% and the immunoreactivity was retained. Both the labelled preparations were tested for bio-distribution in normal and infarcted rats. The activity accumulation in the infarcted region was ∼ 1.5 and 3.5 times as that in normal heart muscle for 125I and 99mTc labelled Mab-4G4 respectively. The major problem with the iodinated antibody was the in vivo deiodination resulting in very high percentage of activity in the thyroid. Although the fraction of the total activity associated with the infarcted heart is not very impressive, the fact that the activities with the infarcted and normal hearths are significantly different is heartening. With further optimisation of labelling and use of F(ab)'2 fragments, better delineation of the infarct sites is aspired. (author)

  13. The development of a human calcitonin radioimunoassay, with 'in house' reagent production, for application to the early diagnosis of medullary thyroid carcioma

    International Nuclear Information System (INIS)

    Reagent production for human Calcitonin (hCT) Radioimmunoassay (RIA) was carried out in our laboratory starting from a kind donation of human synthetic preparation from CIBA (Basel, Switzerland). This product was used for anti-hCT antibody production in rabbits and guinea-pigs and for radioiodination, according to two different methods: classical and stoichiometric Chloramine T techniques. The use of Sephadex G-50 in tracer purification allowed the obtainement of 125I-hCT free of high molecular weight contaminats. A repurification on polyacrylamide gel electrophoresis provided 125I-hCT of higher specific activity that presented specific binginds, to good quality antisera, of the same order of imported tracers (∼ 45%). Different antisera were obtained in rabbits and quinea-pigs, but only one (GP2-IPEN) could be used in such a dilution (1:4000) to provide highly sensitive curves (minimal detectable concentration < 70 pg/ml) presenting, however, very low specific bindings (7-10%). For this reason, in order to be able to set up a regular quality control of our hCT-RIA technique, an antiserum kindly donated by the Karolinska Institute (Stockholm, Sweden) was used. This way, through the use of an higher antibody dilution (1:8000), higher specific bindings (20-30%), higher sensitivies (< 30 pg/ml) and satisfactory precision were obtained. We consider this study a first approach to a complete national production of hCT-RIA reagents, that, at present moment, depends practically only from the obtainement of a high avidity anti-hCT antiserum. More has to be done on accuracy and correct clinical application of this assay to the screening of the familial form of Medullary Thyroid Carcinoma. We also emphasize the fact, due to our limited financial possibilities, all the work was carried out with great economy, taking advantage of previously set up techniques and of the experience already acquired in this field of work. (author)

  14. Synthesis and enantiomeric evaluation of radiolabelled methyl phenidate for the study of dopamine reuptake sites

    International Nuclear Information System (INIS)

    Full text: Neurological diseases such as Parkinson's Disease and associated Movement Disorders have been characterised by reductions in the number of dopamine re-uptake sites or dopamine transporters (DAT) on presynaptic neurons in the striatum. Radiolabelled drugs which display specific and selective binding to the DAT has thus found clinical utility in medical imaging. The psychomotor stimulant methyl phenidate (MP) or Ritalin 1 and several derivatives have demonstrated high affinity binding to these DAT where they exert their pharmacological action. MP possesses two chiral centers with its pharmacological activity being attributed to the dl-threo diastereomer, the dl-erythro being inactive. Furthermore, imaging studies using Positron Emission Tomography (PET) with carbon-11 radiolabelled MP confirmed the d-threo enantiomer to be most active. In vitro studies indicate that compounds substituted with either bromine or iodine in the 3 position of the aromatic ring exhibit high affinity and selective binding to DAT and were thus targeted for radiolabelling with both the PET tracer Bromine-76 as well as the SPECT tracer lodione-123. Threo methyl phenidate and its halogenated derivatives were prepared by the condensation of 3-bromophenylacetonitrile and 2-bromopyridine according to modified literature methods. Radiohalogenation with either 76Br or 123I was achieved by halodestannylation reactions of the corresponding tributyl stannane precursor in the presence of chloramine-T as the oxidant. Purification by C-18 reverse phase HPLC gave the dl-threo product in 70-80% radiochemical yield and in greater than 95% radiochemical purity. Separation of the d and / enantiomers was achieved by chiral HPLC. Biodistribution studies in rodents indicated high uptake of radioactivity in tissues with known DAT sites. PET and SPECT imaging studies in primates indicated high uptake in the striatum compared to the cerebellum reference tissue. The synthesis, diastereomeric separation

  15. 原料奶粉及乳饮料中L-羟脯氨酸的含量测定%Determination of L-hydroxyproline in raw milk powder and milk drink

    Institute of Scientific and Technical Information of China (English)

    陈铭中; 钟旭美; 胡洪森

    2011-01-01

    The method of P-dimethylaminobenzaldehyde spectrophotom etry was used to determine L-hydroxyproline in raw milk powder and milk drink.The result was stable and the recovery rate was high.After hydrolyzed with acid,L-hydroxyproline was released and oxidized by chloramine T.L-hydroxyproline oxide reacted with P-dimethylaminobenzaldehyde and formed red compounds,the absorbance was detected at 580 nm.The method was simple and accurate.Compared with other detecting methods,the expense of it was smaller.It could provide reference for enterprises to detect whether product was adulterated hydrolyzed protein in raw milk powder and milk drink.%采用对二甲氨基苯甲醛分光光度法测定奶粉及乳饮料中L-羟脯氨酸,具有较好的稳定性。奶粉及乳饮料经浓盐酸水解,得到的L-羟脯氨酸经过氯胺T氧化,与对二甲氨基苯甲醛反应生成红色物质,在558nm下测定吸光值。该方法操作简单、准确率高、检测费用相对少,可用于企业奶粉原料掺水解蛋白及产品中是否掺有水解蛋白的检测提供依据。

  16. Identification of the binding subunit of the D/sub 1/-dopamine receptor by photoaffinity crosslinking

    Energy Technology Data Exchange (ETDEWEB)

    Amlaiky, N.; Berger, J.G.; Chang, W.; McQuade, R.D.; Caron, M.G.

    1986-03-01

    A derivative of the potent D/sub 1/ antagonist SCH-23390 has been synthesized. This compound, (R,S)-1-(m-aminophenyl)-7-chloro-8-hydroxy-3-methyl-2,3,4,5-tetrahydro-1H-3-benzazepine (SCH-38548), has been radioiodinated by a chloramine T procedure yielding 3 radioiodinated products. One of these separated isomers (R/sub f/ = 0.35; CH/sub 2/Cl/sub 2/:MEOH:TEA; 82.5:17.5:0.1 on TLC) binds reversibly to rat striatal membranes with high affinity (K/sub D/ approx. 80 pM) appropriate stereoselectivity and D/sub 1/-dopaminergic specificity. (/sup 125/I)SCH-38548 can be covalently incorporated into a peptide of M/sub r/ approx. 72,000 using the heterobifunctional crosslinking reagent N-succinimidyl-6-(4'-azido-2'-nitro-phenylamino) hexanoate. Covalent incorporation of (/sup 125/I) SCH-38548 into the M/sub r/ approx. 72,000 peptide can be blocked by dopaminergic agents with D/sub 1/-dopaminergic specificity (for agonists: SKF 38393 > apomorphine > dopamine; for antagonists: SCH-23390 > cis-flupentixol >>> trans-flupentixol). The D/sub 1/-dopaminergic selectivity and specificity of the labeling was further demonstrated by the fact that other antagonists such as domperidone, ketanserin, phentolamine and alprenolol did not compete for the covalent labeling of the M/sub r/ approx. 72,000 peptide. This new radioligand should be useful in the molecular characterization of the D/sub 1/-dopaminergic receptor from various sources.

  17. Biochemical and pharmacological studies of native and irradiated crotamine with gamma radiation of Co60

    International Nuclear Information System (INIS)

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide from South American rattlesnake venom, composed of 42 amino acid residues. It induces skeletal muscle spasms, leading to a spastic paralysis of hind limbs in mice. The objective was to carry out biochemical and pharmacological studies of native and irradiated crotamine with Co. Crotamine was purified from Crotalus durissus terrificus venom by Sephadex G-100 gel filtration followed by ion exchange chromatography, using a Fast performance Liquid Chromatography (FPLC) system. It was irradiated at 2 mg/ml in 0.15 m NaCl with 2.0 kGy gamma radiation emitted by a Co source. Native and irradiated crotamine were evaluated by biochemical characterization, toxic activity (LD50), and biodistribution. The native and irradiated crotamine were labeled with 29.6 MBq of I using chloramine T method and separated in a Sephadex G-50 column. Male Swiss mice (35 @ 5 g) were injected IP with 0.1 mL (2.4x10 cpm/mouse) of I native crotamine or with 0.4 mL (1.3 x 10 cpm/mouse) of I irradiated crotamine. The animals were sacrificed by ether inhalation at 0.08, 0.25, 0.5,1, 2, 3, 4, 8, 12, and 24 hours. Blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine radioactivity content. The results showed that gamma radiation did not change protein concentration, electrophoretic profile, or protein primary structure, although differences could be seen by spectroscopic techniques. Gamma radiation reduced crotamine toxicity, but did not eliminate bioactivity. Biodistribution studies showed that native and irradiated crotamine have hepatic metabolism and renal elimination. Native and irradiated crotamine have an affinity to skeletal muscle and did not cross the blood-brain barrier. (author)

  18. Therapeutic efficacy of intralesional 131I-labelled hyaluronectin in grafted human glioblastoma

    Energy Technology Data Exchange (ETDEWEB)

    Girard, N.; Courel, M.N.; Vera, P.; Delpech, B. [Centre Henri-Becquerel, Rouen (France). Laboratoire d' Oncologie Moleculaire

    2000-07-01

    The grafted human glioblastoma cell CB109 was used as a model for intralesional therapy with 131I-labelled hyaluronectin glycoprotein (131I-HN). 131I-HN bound specifically to in situ hyaluronic acid (HA), a main component of the extracellular matrix which is involved in tumour invasion. Labelling experimental conditions were determined and, finally, 25 {mu}Ci/{mu}gHN, 1 {mu}g chloramine-T/{mu}gHN and a 60-s stirring period provided a 131I-HN preparation with an optimal affinity for HA (64% compared to unlabelled HN). Following intratumoral injection, 131I-HN was retained with a limited diffusion outside the tumour. On day 4 the radioactivity concentrated in the tumour was still 25 times greater than that in the liver, spleen and kidneys combined. For therapeutic assays, 65 {mu}Ci 131I-HN was injected into the tumour, resulting in a delivery of 6.8 Gy over a 7-day period. Controls received unlabelled HN, heat-inactivated HN, a mixture of inactivated HN plus free 131I or no treatment (six animals per group). Tumour volumes were evaluated every second day from treatment day and the rate of tumour growth was expressed as a ratio of tumour size at time intervals to the tumour size at the time of injection. Growth curves were compared: heat-inactivated with or without free 131I had no anti-tumour effect. Unlabelled HN-injected tumours had a slightly slower growth rate than untreated tumours (p < 0.02) and growth rate of 131I-HN-injected tumours was much lower (p < 0.00002). A pronounced inhibitory effect with intralesional 131I-labelled HN injection resulted from a combination of a) blockage of HA, a proliferation facilitating factor, and b) local irradiation of tumoral tissue, while uptake in normal tissues was minimized.

  19. Harvard--MIT research program in short-lived radiopharmaceuticals

    International Nuclear Information System (INIS)

    This report describes progress on five projects. The first project showed a 1000 fold concentration of the cationic complex 99mTc (MIBI) in heart cell mitochondria vs heart cell cytoplasm, as determined by high resolution electron probe microanalysis. Additional technetium-99m based complexes are being developed and tested. The second project involves evaluating technetium acetylacteonates as potential indicators of cerebral blood flow. An intermediate in the synthesis of a technetium porphyrin complex has been synthesized; an oxotechnetium(V)-2,4-pentanedione complex has been prepared and is currently being characterized. The third project involves using radio labelled antibodies for diagnosis and treatment of cancer. An early discovery was that chloramine-T based iodination protocols resulted in a reversal of the charge on mouse lgGs. Immunoperoxidase-labelled monoclonal antibody MOv 18 was shown to bind specifically to the most frequent ovarian aderon carcinomas, and not to healthy tissue, making this antibody a good candidate for immunotherapy or immunodetection. Work on a specific immunotherapy protocol suffered a setback when one reagent, a 125I-biotin complex, proved to be unstable in vivo. The fourth project involves labelling antibodies with positron emitting radionuclides. Radiofluorination was accomplished through reductive alkylation of 18F-aldehyde, or pentafluorophenyl esters. Radioiodination was accomplished using alkyl-tin derivation exchange. The fifth project examined antibody modification for use in radioimmune imaging. Technetium-99m-labelled lgG was shown to be biologically equivalent to Indium-III-labelled lgG for imaging focal sites of inflamation. Also, Indium III labelling of small bioactive peptides was examined as a means of imaging important physiological processes. 44 refs., 2 figs

  20. Radioimmunotherapy in refractory b-cell nonhodgkins lymphoma with I-131-labeled chimeric anti cd-20 c2b8 (I-131 rituximab): preliminary result

    Energy Technology Data Exchange (ETDEWEB)

    Kang, Hye Jin; Park, Yeon Hee; Kim, Sung Eun and others [Korea University Medical School, Seoul (Korea, Republic of)

    2005-07-01

    Recently, the native chimeric human-mouse anti CD-20 antibody IDEC-C2B8 (Rituximab) has been widely applied in NHL. This ongoing phase study was to evaluate whether radioimmunotherapy (RIT) with I-131 rituximab is effective in refractory B-cell NHL. Inclusion criteria were as follows: B-cell NHL with relapsed or refractory to primary standard therapy, measurable disease, adequate hematologic, renal, and hepatic function, informed consent. The rituximab (Mabthera, Roach) was radiolabeled with iodine-131(I-131) using a modified chloramine T method with high radiochemical purity (95%) and preservation of immuno-reactivity. All patients received loading doses of unlabeled rituximab (median, 40 mg: range, 20{approx}70 mg) immediately prior to administration of therapeutic dose (51.4{approx}152.2 MBq/kg), and then underwent gamma camera scan. 11 patients were enrolled (4 low-grade B-cell NHL, 7 DLBCL, median age 63 years). Patients had received a median of three prior chemotherapy regimens. The objective response rate was 36.4% (1 CR, 3 PRs). These all responses were observed in low-grade B-cell NHL, except one with DLBCL. Adverse events were primarily hematologic toxicities; the incidence of grade 3/4 neutropenia, thrombocytopenia, and anemia was 27.3%, 45.5%, and 18.2%, respectively. The treatment-related mortality was observed in one patient, who had been previously treated with high-dose chemotherapy plus TBI with autologous stem cell transplantation. RIT with I-131 rituximab seems to be effective tolerable in refractory low-grade B-cell NHL, although modest activity in refractory DLBCL. Further studies to define the efficacy of I-131 rituximab in DLBCL are warranted.

  1. Pharmacokinetics of Pegylated recombinant new human Interleukin-6 (PET-rhIL-6)

    International Nuclear Information System (INIS)

    In this study, the newly developed Pegylated recombinant human Interleukin-6 (PEG-rhIL-6) was investigated. With 125I labeled PEG-rhIL-6 by two-phase chloramine-T method. Thirty of the SD rats were randomly divided into 6 groups (group 1, 2, 3, 4, 5 and 6, respectively), with 5 animals (3 males and 2 females) each group. Group 1, 2 and 3 were injected subcutaneously with 125I-PEG-rhIL-6 at the dose of 40, 20 and 3 μg/kg, respectively. Group 4, 5 and 6 were injected intravenously with 125I-PEG-rhIL-6 with the same dose level, respectively. Blood samples were collected at different time intervals after injection. The plasma concentration (ng/ml) of 125I-PEG-rhIL-6 were determined by 125I isotope tracing method and then assessed with 3P87 software to obtain the pharmacokinetic parameters. Results clearly showed that 1/2 ke (20.41, 21.53, 19.77) and tl/2 ka (10.44, 11.26, 11.37) of group 1-3 are higher number than t1/2β (17.54, 18.78, 17.81) and t1/2α (1.93, 1.60, 1.78) of group 4-6. The pharmacological activity of 125I-PEG-rhIL-6 maintain longer time by subcutaneous injection; The pharmacokinetic of V/Fc after sc (694.96 ml/kg) is higher than those after iv (115.26 ml/kg), which showed that the excretion speed of 125-PEG-rhIL-6 given by sc was slowly. The above results obtained indicated that the pharmacokinetic characteristics of 125I-PEG-rhIL-6 would provide important information for clinical application and the subcutaneous injection is better. (authors)

  2. Role of the metabolism of parathyroid hormone

    International Nuclear Information System (INIS)

    The heterogeneity of parathyroid hormone (PTH) in plasma has prompted investigations of the metabolism of PTH and its relationship to hormone action. The time course of tissue distribution and metabolism of electrolytically iodinated PTH (E-PTH) previously shown to retain biological activity was compared with that of inactive PTH iodinated with Chloramine-T (CT-PTH). Labeled PTH (0.4 μg) was injected in the saphenous veins of anesthetized rats which were sacrificed at 1, 3, 5, 10, and 20 min. Tissue extracts from kidney, liver, and serum were chromatographed to separate intact PTH from its metabolites. In the kidney, the initial rate of degradation of E-PTH was greater than that of CT-PTH. The difference in initial rates of metabolism may be due, in part, to receptor-specific hydrolysis on peritubular cell membranes which selectively act on biologically active PTH molecules. PTH-responsive adenyl cyclase activity in isolated kidney cortex plasma membranes was measured and PTH metabolism was monitored simultaneously. When degradation was completely blocked by histone f3 (1 mg/ml), adenyl cyclase activity was significantly increased over control. In addition, when adenyl cyclase activity was negligible, the rate of PTH degradation by the membranes was not significantly diminished. Consistent with the in vivo data was the observation that E-PTH is metabolized by these membranes at a greater rate than CT-PTH. The data demonstrate the existence of a receptor-specific metabolism at sites which are independent of PTH receptor mediated adenyl cyclase activity

  3. Development of anti-CD30 radioimmunoconstructs (RICs) for treatment of Hodgkin's lymphoma. Studies with cell lines and animal studies

    International Nuclear Information System (INIS)

    Objectives: comparison of the binding affinity to a CD30-positive Hodgkin lymphoma (HL) cell line and biodistribution in HL bearing mice of new anti-CD30 radioimmunoconjugates (RICs) of varying structure and labelling nuclides. Methods: The antibodies Ki-4 and 5F11 were radioiodinated by the chloramine T method or labelled with 111In via p-NCS-Benzyl-DOTA. In addition, the Ki-4-dimer was investigated in the iodinated form. The RICs were analyzed for retained immunoreactivity by immunochromatography. In-vitro binding studies were performed on CD30-positive L540 cell lines. For in-vivo biodistribution studies, SCID mice bearing human HL xenografts were injected with the various radio-immunoconjugates. After 24 h, activities in the organs and tumour were measured for all 5 RICs. Tumour-free animals were studied in the same way with 131I-Ki-4 24 h p. i. The three RICs with the highest tumour/background ratios 24 h p.i. (131I-Ki-4, 131I-5F11, 111In-bz-DOTA-Ki-4) were analysed further at 48 h and 72 h. Results: all the RICs were successfully labelled with high specific activities (28-47 TBq/mmol) and sufficient radiochemical yields (> 80%). Scatchard plot analysis proved high tumour affinity (KD = 20-220 nmol/l). In-vivo tumour accumulation in % of injected dose per g tissue (% ID/g) lay between 2.6 (131I-5F11) and 12.3 % ID/g (131I-Ki-4) with permanently high background in blood. Tumour/blood-ratios of all RICs were below one at all time points. Conclusions: in-vitro tumour cell affinities of all RICs were promising. However, in-vivo biokinetics tested in the mouse model did not meet expectations. This highlights the importance of developing and testing further new anti-CD30 conjugates. (orig.)

  4. A radioimmunoassay detecting the borine leukaemia virus transmembrane protein gp30 and anti-gp30 antibodies in the serum of cattle

    International Nuclear Information System (INIS)

    From virus-infected fetal lamb kidney (FLK) cells a relatively homogeneous envelope transmembrane protein gp30 of bovine leukemia virus (BLV) was isolated. As shown by a partial sequence analysis of the N-terminus of this protein, the gp30 preparation contained only traces (less than 5%) of p24 gag protein: rabbit anti-gp30 serum did not cross react with the BLV proteins gp51, p12, p151, p152 and p10 but reacted weakly with the p24 polypeptide. 125I-labelled gp30 (chloramine-T) was precipitated with the serum of BLV-infected cattle. Nonlabelled preparation of gp30 competitively inhibited the reaction of 125I-labelled gp30 with natural antibodies. 193 cattle sera were investigated by liquid phase radioimmunoassay (RIA) using 125I-gp30, gp51 and p24 antigens. Sixteen noninfected cattle sera were negative in all tests. The 177 serum samples of BLV-infected animals were examined to the diagnostic value of the three tests. Of these, 175 were positive in gp51 RIA, 172 in p24 RIA and 164 in gp30 RIA. In all three tests, 159 sera were positive while 18 sera, mostly coming from animals with normal leukocyte counts, were positive only either with gp51 or p24, or were double positive with either gp51/p24 or gp51/gp30. It is concluded that gp51 RIA is superior to both gp30 and p24 RIA and that gp30 RIA will be useful for investigating the role of gp30 in virus pathogenicity. (author). 2 figs., 2 tabs., 33 refs

  5. Fast determination of cyanide by paper disk method%氰化物纸片快速测定食品中的氰化物

    Institute of Scientific and Technical Information of China (English)

    杨俊; 陈亿展; 孟令兵; 赖小丽; 李意

    2012-01-01

    目的:建立食品中氰化物的纸片速测法并应用于重大活动保障中.方法:食品中的氰化物经水浸取后,与氯胺T反应生成氯化氰,继而与异烟酸反应,并经水解而成戊烯二醛,最后与1,3-二甲基巴比妥酸作用生成紫蓝色化合物,比色定量.结果:方法检出限为1.00 mg/kg,准确度和精密度都较为理想.结论:氰化物纸片速测法具有灵敏度高,快速,携带方便的特点,适合于重大活动保障中氰化物的快速测定.%Objective:To establish a method for fast determination of cyanide in food by paper disk method in important Activities. Methods: Cyanide in food was leached with water,and then reacted with chloramine T to form ey-anochloride which reacted with isonicotinic acid, the product was hydrolyed to glutaconaldehyde, and finally it reacted with 1,3- dimethyl barbituric acid to form a violet blue compound. The content of cyanide was quantitated colorime-trically. Results: The detection limit of the method was 1. 00 mg/kg, the recoveries and the precision were satisfactory. Conclusion: The method was sensitive, fast and convenient. It is suitable to determine cyanide in food in important Activities.

  6. Role of the metabolism of parathyroid hormone. [Rats

    Energy Technology Data Exchange (ETDEWEB)

    Teitelbaum, Anne P.

    1978-01-01

    The heterogeneity of parathyroid hormone (PTH) in plasma has prompted investigations of the metabolism of PTH and its relationship to hormone action. The time course of tissue distribution and metabolism of electrolytically iodinated PTH (E-PTH) previously shown to retain biological activity was compared with that of inactive PTH iodinated with Chloramine-T (CT-PTH). Labeled PTH (0.4 ..mu..g) was injected in the saphenous veins of anesthetized rats which were sacrificed at 1, 3, 5, 10, and 20 min. Tissue extracts from kidney, liver, and serum were chromatographed to separate intact PTH from its metabolites. In the kidney, the initial rate of degradation of E-PTH was greater than that of CT-PTH. The difference in initial rates of metabolism may be due, in part, to receptor-specific hydrolysis on peritubular cell membranes which selectively act on biologically active PTH molecules. PTH-responsive adenyl cyclase activity in isolated kidney cortex plasma membranes was measured and PTH metabolism was monitored simultaneously. When degradation was completely blocked by histone f/sub 3/ (1 mg/ml), adenyl cyclase activity was significantly increased over control. In addition, when adenyl cyclase activity was negligible, the rate of PTH degradation by the membranes was not significantly diminished. Consistent with the in vivo data was the observation that E-PTH is metabolized by these membranes at a greater rate than CT-PTH. The data demonstrate the existence of a receptor-specific metabolism at sites which are independent of PTH receptor mediated adenyl cyclase activity.

  7. Preparation of mono-radioiodinated tracers for study of the in vivo metabolism of atrial natriuretic peptide in humans

    Energy Technology Data Exchange (ETDEWEB)

    Clerico, A. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Iervasi, G. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Manfredi, C. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Salvadori, S. [Dept. of Pharmaceutical Sciences, Univ. of Ferrara (Italy); Marastoni, M. [Dept. of Pharmaceutical Sciences, Univ. of Ferrara (Italy); Del Chicca, M.G. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Giannessi, D. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Del Ry, S. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Andreassi, M.G. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Sabatino, L. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Iascone, M.R. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Biagini, A. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica; Donato, L. [Consiglio Nazionale delle Ricerche, Pisa (Italy). Lab. di Fisiologia Clinica

    1995-09-01

    The authors evaluate the optimum chemical conditions for labelling atrial natriuretic peptide (ANP) and its metabolites and for preparing highly purified radiotracers which can be used for in vivo kinetic studies of ANP in humans. Synthetic {alpha} h{sub 1-28}ANP and some hormone metabolites were iodinated with Na{sup 125}I or Na{sup 131}I by means of the lactoperoxidase (ANP) or the chloramine-T (ANP metabolites) technique. The biological activity of labelled ANP was tested by means of a binding study using mouse cardiac membranes. A high-performance liquid chromatography (HPLC) procedure was used to purify the labelled hormone and the principal labelled metabolites in venous plasma samples collected up to 50 min after the injection of {sup 125}I-labelled ANP from nine healthy men. The main ANP kinetic parameters were derived from the disappearance curves of the [{sup 125}I]ANP, which were satisfactorily fitted by a bi-exponential function in all subjects. The main advantages of this tracer technique are high accuracy, allowing the identification of the metabolites produced in vivo under steady-state conditions after injection of the precursor (labelled hormone) high sensitivity, allowing the detection of minimal quantities of metabolites high specificity, allowing the detection of possible in vitro artefactual generation of cleavage products of ANP using an internal labelled standard. Utilizing this tracer method, it was possible to estimate the principal parameters of ANP kinetics and also to plot the appearance curves of the labelled metabolites produced in vivo after the injection of the labelled precursor. (orig.). With 5 figs.

  8. Blood-brain barrier transport of cationized immunoglobulin G: Enhanced delivery compared to native protein

    Energy Technology Data Exchange (ETDEWEB)

    Triguero, D.; Buciak, J.B.; Yang, J.; Pardridge, W.M.

    1989-06-01

    IgG molecules are potential neuropharmaceuticals that may be used for therapeutic or diagnostic purposes. However, IgG molecules are excluded from entering brain, owing to a lack of transport of these plasma proteins through the brain capillary wall, or blood-brain barrier (BBB). The possibility of enhanced IgG delivery through the BBB by cationization of the proteins was explored in the present studies. Native bovine IgG molecules were cationized by covalent coupling of hexamethylenediamine and the isoelectric point was raised to greater than 10.7 based on isoelectric focusing studies. Native and cationized IgG molecules were radiolabeled with /sup 125/I and chloramine T. Cationized IgG, but not native IgG, was rapidly taken up by isolated bovine brain microvessels, which were used as an in vitro model system of the BBB. Cationized IgG binding was time and temperature dependent and was saturated by increasing concentrations of unlabeled cationized IgG (dissociation constant of the high-affinity binding site, 0.90 +/- 0.37 microM; Bmax, 1.4 +/- 0.4 nmol per mg of protein). In vivo studies documented enhanced brain uptake of 125I-labeled cationized IgG relative to (3H)albumin, and complete transcytosis of the 125I-labeled cationized IgG molecule through the BBB and into brain parenchyma was demonstrated by thaw-mount autoradiography of frozen sections of rat brain obtained after carotid arterial infusions of 125I-labeled cationized IgG. These studies demonstrate that cationization of IgG molecules greatly facilitates the transport of these plasma proteins through the BBB in vivo, and this process may provide a new strategy for IgG delivery through the BBB.

  9. Preparation of mono-radioiodinated tracers for study of the in vivo metabolism of atrial natriuretic peptide in humans

    International Nuclear Information System (INIS)

    The authors evaluate the optimum chemical conditions for labelling atrial natriuretic peptide (ANP) and its metabolites and for preparing highly purified radiotracers which can be used for in vivo kinetic studies of ANP in humans. Synthetic α h1-28ANP and some hormone metabolites were iodinated with Na125I or Na131I by means of the lactoperoxidase (ANP) or the chloramine-T (ANP metabolites) technique. The biological activity of labelled ANP was tested by means of a binding study using mouse cardiac membranes. A high-performance liquid chromatography (HPLC) procedure was used to purify the labelled hormone and the principal labelled metabolites in venous plasma samples collected up to 50 min after the injection of 125I-labelled ANP from nine healthy men. The main ANP kinetic parameters were derived from the disappearance curves of the [125I]ANP, which were satisfactorily fitted by a bi-exponential function in all subjects. The main advantages of this tracer technique are high accuracy, allowing the identification of the metabolites produced in vivo under steady-state conditions after injection of the precursor (labelled hormone) high sensitivity, allowing the detection of minimal quantities of metabolites high specificity, allowing the detection of possible in vitro artefactual generation of cleavage products of ANP using an internal labelled standard. Utilizing this tracer method, it was possible to estimate the principal parameters of ANP kinetics and also to plot the appearance curves of the labelled metabolites produced in vivo after the injection of the labelled precursor. (orig.). With 5 figs

  10. {sup 123}I-BPA and {sup 123}I-BPA-fructose complex as a new radiopharmaceutical for the imaging of amino acid transport in tumor

    Energy Technology Data Exchange (ETDEWEB)

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

    1999-07-01

    Boronophenylalanine (BPA) as derivative of phenylalanine, was used to treatment for glioma and melanoma in BNCT. We labeled BPA with radioiodides for tumor imaging of amino acid transport with gamma camera. Because of limited solubility of BAP, I-BPA-Fructose(I-BPA was complexed with fructose) to increase solubility. I-BPA was labeled by chloramine T coated bead method. Serum stability of I-BPA analyzed by HPLC at 37 .deg. C. Cellular uptake of I-BPA and I-BPA-Fructose was compared in 9L glioma and B16 melanoma. To see biodistribution, I-BPA 9x10{sup 5}Bq(20 {mu}g/100 {mu}l) or I-BPA-Fructose 9x10{sup 5}Bq(20 {mu}g/fructose 55 {mu}g/100 {mu}l) was injected to B16 melanoma bearing C57 mice. In tumor bearing mice at 30 min, 1, 2, 24 hr after injection of tracers (n-4 per group). In 24hr, radiochemical purity of I-BPA in serum was retained above 90%. In cultured cells the maximum uptake was observed at 60min. In 9L glioma cells, %uptake of I-BPA and I-BPA-Fructose was 2.05, 2.6 at 60min. But in B16 melanoma, %uptake of I-BPA and I-BPA and-Fructose was 2.57, 6.62 at 60 min. In melanoma bearing mice, tumor/muscle ratio of I-BPA in 30 min, 1hr, 2hr, 24hr after injection was 1.48, 2.19, 2.28, 0.29 and %ID/g of tumor was 6.25, 5.17, 3.52, 0.29. Tumor/muscle ratio of I-BPA-Fructose was 1.51, 2.05, 2.1, 2.84 in 30 min, 1hr, 2hr, 24hr post-injection and %ID/g of tumor was 4.61, 3.65, 2.93, 0.71. The radioactivity was excreted mainly via hepatobiliary tract to the intestine. I-BPA was stable in serum upto 24hr. Uptake of I-BPA-Fructose was higher than I-BPA in melanoma cells. I-BPA-Fructose, is a promising tumor imaging radiopharmaceutical in some tumors.

  11. Biochemical and pharmacological studies of native and irradiated crotamine with gamma radiation of {sup 60}Co; Estudo bioquimico e farmacologico das crotaminas nativa e irradiada com radiacao gama de {sup 60}Co

    Energy Technology Data Exchange (ETDEWEB)

    Mitake, Malvina Boni

    2000-07-01

    Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide from the South American rattlesnake venom, composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of this thesis was carry out biochemical and pharmacological studies of native and irradiated crotamine with {sup 60} Co. Crotamine was purified from Crotalus durissus terrificus venom by Sephadex G-100 gel filtration followed by ion exchange chromatography, using a Fast Performance Liquid Chromatography (FPLC) system. It was irradiated at 2 mg/ml in 0.15 M Na Cl with 2.0 kGy gamma radiation emitted by a {sup 60} Co source. The native and irradiated crotamine were evaluated by biochemical characterization, toxic activity (LD{sub 50} and biodistribution. The native and irradiated crotamine were labelled with 29.6 MBq of {sup 125} I using chloramine T method, and separated in a Sephadex G-50 column. Male Swiss mice (35{+-} 5 g), were injected i.p. with o.1 mL (2.4 x 10{sup 6} cpm/mouse) of {sup 125} I native crotamine or with 0.4 mL (1.3 x 10{sup 6} cpm/mouse) of {sup 125} I irradiated crotamine. At 0.08; 0.25; 0.5; 1; 2; 4; 8; 12 and 24 hours the animal were killed by ether inhalation. Blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine radioactivity content. The results showed that gamma radiation did not change the protein concentration, the electroforetic profile or the primary structure of the protein, although differences were shown by spectroscopic techniques. The gamma radiation diminished the toxicity of crotamine, but it did not abolish bioactivity. Biodistribution studies showed that native and irradiated crotamine have hepatic metabolism and renal elimination. The native and irradiated crotamine have affinity by skeletal

  12. Development of anti-CD30 radioimmunoconstructs (RICs) for treatment of Hodgkin's lymphoma. Studies with cell lines and animal studies

    Energy Technology Data Exchange (ETDEWEB)

    Dietlein, M.; Boerner, S.M.; Fischer, T.; Zimmermanns, B.; Kobe, C.; Schicha, H.; Schomaecker, K. [Dept. of Nuclear Medicine, Univ. of Cologne (Germany); Hansen, H.; Schnell, R.; Tawadros, S.; Engert, A.; Staak, O.; Pogge von Strandmann, E. [Dept. of Internal Medicine I, Univ. of Cologne (Germany)

    2010-07-01

    Objectives: comparison of the binding affinity to a CD30-positive Hodgkin lymphoma (HL) cell line and biodistribution in HL bearing mice of new anti-CD30 radioimmunoconjugates (RICs) of varying structure and labelling nuclides. Methods: The antibodies Ki-4 and 5F11 were radioiodinated by the chloramine T method or labelled with {sup 111}In via p-NCS-Benzyl-DOTA. In addition, the Ki-4-dimer was investigated in the iodinated form. The RICs were analyzed for retained immunoreactivity by immunochromatography. In-vitro binding studies were performed on CD30-positive L540 cell lines. For in-vivo biodistribution studies, SCID mice bearing human HL xenografts were injected with the various radio-immunoconjugates. After 24 h, activities in the organs and tumour were measured for all 5 RICs. Tumour-free animals were studied in the same way with {sup 131}I-Ki-4 24 h p. i. The three RICs with the highest tumour/background ratios 24 h p.i. ({sup 131}I-Ki-4, {sup 131}I-5F11, {sup 111}In-bz-DOTA-Ki-4) were analysed further at 48 h and 72 h. Results: all the RICs were successfully labelled with high specific activities (28-47 TBq/mmol) and sufficient radiochemical yields (> 80%). Scatchard plot analysis proved high tumour affinity (K{sub D} = 20-220 nmol/l). In-vivo tumour accumulation in % of injected dose per g tissue (% ID/g) lay between 2.6 ({sup 131}I-5F11) and 12.3 % ID/g ({sup 131}I-Ki-4) with permanently high background in blood. Tumour/blood-ratios of all RICs were below one at all time points. Conclusions: in-vitro tumour cell affinities of all RICs were promising. However, in-vivo biokinetics tested in the mouse model did not meet expectations. This highlights the importance of developing and testing further new anti-CD30 conjugates. (orig.)

  13. Development of a new radioimmunoassay for erythropoietin using recombinant erythropoietin

    Energy Technology Data Exchange (ETDEWEB)

    Mason-Garcia, M.; Beckman, B.S.; Brookins, J.W.; Powell, J.S.; Lanham, W.; Blaisdell, S.; Keay, L.; Li, S.C.; Fisher, J.W. (Tulane Univ. School of Medicine, New Orleans, LA (USA))

    1990-11-01

    The development of a 24 hour radioimmunoassay for erythropoietin (EPO) using EPO derived from recombinant DNA as both immunogen and ligand is described in the present paper. Mixed breed rabbits immunized with 10 micrograms/kg of EPO derived from a stably transfected cell line (MD) produced antibodies to EPO with high titer (up to 1:896,000 final dilution in the tube), high affinity (8.4 x 10(11) liter/M), and good specificity. Purified EPO from the above source or from AmGen Biologicals (AG) were successfully radioiodinated with the chloramine-T method and used as ligand in the radioimmunoassay. Standard dose-response curves prepared with EPO from both commercial sources were not significantly different and showed a sensitivity of 0.75 to 0.96 mU/tube. The dose-response curves in both systems also showed parallelism with serially diluted serum from a patient with aplastic anemia. Within-assay and between-assay precision were determined by assaying multiple replicates of a serum pool. Recovery of exogenous EPO added to a serum pool averaged 97% for both systems. The range of normal human serum EPO was determined by assaying the sera of 153 hematologically-normal adult subjects and was found to be 1.1 to 27.3 mU/ml for MD EPO and 0.5 to 16.7 mU/ml for AG EPO. Sera from several patients with hematologic abnormalities were also assayed, including those of 36 patients with anemia of end-stage renal disease (mean +/- SEM, 29.5 +/- 4.0 mU/ml; P less than 0.01). In conclusion, this new, more rapid and sensitive radioimmunoassay system can be used to measure EPO levels in sera from normal human subjects and patients with several types of anemia, and should also be very useful in therapeutic drug monitoring of patients receiving EPO from various commercial sources.

  14. Immunoassay of taxol using anti-taxol antibody and radiolabeled taxol

    Energy Technology Data Exchange (ETDEWEB)

    Lee, T. S.; Hong, J. P.; Kim, H. S.; Awh, O. D.; Choi, C. W.; Lim, S. M. [College of Medicine, Yonsei Univ., Wonju (Korea, Republic of)

    2000-07-01

    Taxol (Paclitaxel), an antineoplastic agent, has been used for treatment of ovarian and breast cancer. Prompt analysis of taxol concentration in the human serum is beneficial to maintain an appropriate taxol concentration in blood, which will enhance therapeutic effect of the drug. This study was aimed to establish a radioimmunoassay system for monitoring the taxol level in blood during cancer treatment. Hemisuccinyltaxol (HT) was synthesized by esterification of taxol with succinic anhydride. Tyraminehemisuccinyltaxol (THT) was synthesized by amide bond formation of the hemisuccinyltaxol with tyramine in the presence of isobutylchloroformate. The synthesized THT was purified by HPLC, Tyraminehemisuccinyltaxol was dissolved in 80% acetonitrile solution, which was stored at 4 .deg. C or 37 .deg. C for 7 days. The stability of tyraminehemisuccinyltaxol was monitored during the storage period. Tyramine-hemisuccinyltaxol was labeled with {sup 125}I (3.7 x 10{sup 7} Bq) by Chloramine-T. The radiolabeled product, ({sup 125}I)iodo-tyraminehemiscuccinyltasol, was allowed to stand at 4 .deg. C or 37 .deg. C for up to 7 days to estimate its stability. The radiochemical purity of ({sup 125}I) iodotyramine-hemisuccinyltaxol was determined by HPLC. In other hand, the titer of taxol monoclonal antibody (3G5A7) was determined by radioimmunoassay using ({sup 125}I) iodotyramine-hemisuccinyltaxol as a radiolabeled antigen. A standard dose response curve was plotted by taxolcompetitive radioimmunoassay. Hemisuccinyltaxol was synthesized with 79.9% yield. Tyraminehemisuccinyltaxol was synthesized with 19.5% yield and purified by HPLC. the chemical purity of Tyraminehemisuccinyltaxol was maintained at 96.5% and 97.5% after 7 days storage at 4 .deg. C and 37.deg. C respectively. The radiochemical purity of ({sup 125}I) iodotyramine-hemisuccinyltaxol was diminished to 88.1% and 86.2% after 7 days storage at 4 .deg. C and 37. deg.C, respectively. The antibody titer against taxol was 1

  15. Preparation of {sup 125}I-iodotyraminehemisuccinyltaxol ({sup 125}ITHT) for competitive taxol radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Tae Sup; Awh, Ok Doo; Choi, Tae Hyun; Kim, Hyun Suk; Hong, Jun Pyo; Lee, Eun Sook [College of Medicine, Yonsei Univ., Wonju (Korea, Republic of); Choi, Chang Woon; Lim, Sang Moo [Korea Cancer Center Hospital, Seoul (Korea, Republic of)

    2002-04-01

    Taxol(Paclitaxel), an antineoplastic agent, has been used in the treatment of ovarian and breast cancers. The determination of optimal Taxol concentrations in human serum was required for enhancing therapeutic effect and maintaining the appropriate Taxol level in blood. This study was aimed to synthesize radiolabeled Taxol derivatives as radiotracer in competitive radioimmuoassay for monitoring Taxol concentrations in blood and to determine the usefulness of its derivatives. Hemisuccinyltaxol(HT) was synthesized by esterification of Taxol with succinic anhydride. Tyraminehemisuccinyltaxol (THT) was synthesized by coupling of HT with tyramine using isobutylchloroformate as coupling agent and purified by HPLC. By using chloramine-T(5.25 mg/ml, 10 {mu}l) as oxidant agent, THT (4 mg/ml, 30 {mu}l) was labeled with {sup 125}I (37 MBq, 1 mCi). To estimate the stability of purified THT, {sup 125}I-iodotyraminehemisuccinyltaxol ({sup 125}ITHT) was dissolved in 80% acetonitrile aqueous solution, and the solution was incubated at 4 .deg. C and 37 .deg. C for 7 days. At various time intervals, the stability of THT and {sup 125}ITHT was monitored. The titer of Taxol monoclonal antibody, 3G5A7, was determined by competitive radioimmunoassay using {sup 125}ITHT as a labeled antigen. A standard dose-response curve was demonstrated by Taxol competitive radioimmunoassay. HT and THT were synthesized with 79.9% and 19.5% yield, respectively. The labeling yield of {sup 125}ITHT was 93%. After 7 days, the chemical purity of THT was 96.5% at 4 .deg. C, and 97.5% at 37 .deg. C. After 3 days, {sup 125}ITHT was stable with 94.7% at 4 .deg. C and 93.4% at 37 .deg. C. After 7 days radiochemical purity was diminished to 88.1% at 4 .deg. C and 86.1% at 37 .deg. C. The titer of Taxol monoclonal antibody, 3G5A7, was determined to 1:256. A standard dose-response cure demonstrated good collinearity (R{sup 2}=0.971) as Taxol concentration-dependent manner. Competitive radioimmunoassay using {sup 125

  16. The in vivo pharmacokinetics study of anti-ProGRP(31-98) monoclonal antibody E-B5

    International Nuclear Information System (INIS)

    Objective: Progastrin-releasing peptide(31-98) (ProGRP(31-98) is a specific tumor marker in patients with small cell lung cancer (SCLC). E-B5 antibody against ProGRP(31-98) was produced by China Institute for Radiation Protection. The aim of this study was to explore the 131I labeling methods, stability, immunological activity and biological distribution pattern of E-B5 antibody against ProGRP(31-98). Methods: Chloramine-T method was used for 131I labeling E-B5 antibody. Labeling efficiency, radiochemical purity and stability were estimated by using paper chromatography method. Immunological activity of 131I-E-B5 was detected with cell conjugation assay. After healthy Kunming mice were injected with 131I-E-B5 antibody through tail veil, the biodistribution and pharmacokinetics of 131l-E-B5 antibody in healthy Kunming mice were studied. Continuous images of the nude mice beating SCLC were carried out at different time points after injection of 131I-E-B5 antibody. Continuous variables were expressed as x-bar ± s and compared by t-test with SPSS 13.0 software. Results: The labeling efficiency and radiochemical purity of 131I-E-B5were 90.8%, 99.28%, respectively. The radiochemical purity still maintained above 70% after incubation in water bath at 37 degree C for 24 h. After incubation with healthy serum for 24 h, the radiochemical purity still reached 68.1%. The immunobinding ratios were 71.6% and 33.2% for NCI-H446 cells and A549 cells respectively. The in vivo distribution and elimination of 131I-E-B5 antibody were consistent with a first-order and two-compartment model, t1/2α =0.2 h, t1/2β = 8.35 h. The metabolism of 131I-E-B5 antibody mainly depended on liver and kidney and with rapid elimination in blood. Conclusions: 131I-E-B5 antibody not only has high labeling efficiency and radiochemical purity, but also has good stability and keeps good immunological activity. 131I-E-B5 is a promising agent of radioimmunoimaging and radioimmunotherapy of SCLC. (authors

  17. Vasoactive intestinal peptide receptor scintigraphy in patients with pancreatic adenocarcinomas or neuroendocrine tumours

    International Nuclear Information System (INIS)

    Human adenocarcinomas of the gastroenteropancreatic system overexpress vasoactive intestinal peptide (VIP) receptors and therefore represent logical diagnostic targets for receptor scintigraphy. Using iodine-123 labelled VIP, the newly employed diagnostic procedure termed VIP receptor scintigraphy (VIP-RS) appears to detect tumour tissue, especially pancreatic metastatic tumours, in almost all cases. So far, however, only a single centre has demonstrated convincing positive results. The aim of this study was to compare the sensitivity and specificity of VIP-RS with those of computer tomography (CT) and transabdominal ultrasound in patients with extensive pancreatic metastatic adenocarcinomas and neuroendocrine tumours. VIP was radiolabelled with carrier-free 123I using the chloramine T-method and preparative high-performance liquid chromatography for purification. Patients with metastatic pancreatic (n=12) and colorectal (n=3) carcinomas (adenocarcinoma: n=13, neuroendocrine tumour: n=2) were studied by VIP-RS, CT, ultrasound and, in one case, also by radioligand receptor autoradiography. Carrier-free radioiodinated VIP of maximum specific radioactivity maintained a high biological activity as determined by cAMP formation in receptor-expressing tumour cell lines. Intravenous injection of 123I-VIP did not cause any side-effects. Biodistribution, determined over 24 h, was high in the lungs and low in abdominal organs. Although all patients had extensive metastatic disease as evidenced by CT and ultrasound, VIP-RS was unable to detect either primaries or metastases in these patients. Only in two patients could a significant uptake of radiolabel be detected in organs directly infiltrated by the primary. To exclude false-negative findings, tumour tissue in one patient with a large primary, undetectable by VIP-RS, was analysed by radioligand receptor autoradiography and shown to be receptor positive. Moreover, in vitro receptor determinations showed that pancreatic

  18. Distribution of 131I-labeled Bothrops erythromelas venom in mice

    Directory of Open Access Journals (Sweden)

    C.M.L. Vasconcelos

    1998-03-01

    Full Text Available Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131I by the chloramine T method, and separated in a Sephacryl® S-200 column. The efficiency of labeling was 68%. Male Swiss mice (40-45 g, which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc with 0.3 ml (2.35 x 105 cpm/mouse of 131I-venom (N = 42, 131I-albumin or 131I (controls, N = 28 each. Thirty minutes and 1, 3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% NaCl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption in the skin (51% within the first 30 min compared to albumin (20.1% and free iodine (8.2%. Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach (3rd h, small intestine (3rd h and large intestine (6th h. Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively, and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3rd h postinjection (1.98%. There was a low binding of labeled venom in the liver (<0.54%, thyroid (<0.11% and lungs (<0.08%, and no iodinated venom was detected in brain, heart, diaphragm, spleen or bladder. The low venom binding observed in most internal organs, comparable to that of albumin, suggests that B. erythromelas venom does not specifically target most internal organs

  19. Distribution of 131 I- labeled Bothrops erythromelas venom in mice

    International Nuclear Information System (INIS)

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with 131 I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 105 cpm/mouse) of 131 I-venom (N = 42), 131 -albumin or 131 I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs. That is, the systemic effects of envenomation ar mainly due to an indirect action. (author)

  20. Comparative Studies on the Radiolabeling and Chromatographic Purification of Some Medically Important Compounds

    International Nuclear Information System (INIS)

    The present thesis comprises five basic chapters: The first chapter includes the main idea of our study and its problems, also includes the aim of the work of our study. The second chapter includes the theoretical consideration of the subject. It deals with the general methods of labeling, factors that influence the integrity of labeled compounds, radionuclides used for diagnostic nuclear medicine, production methods and radioactive properties of 123I, 125I and 131I. It includes also the techniques used for the preparation of the radioiodinated compounds especially the electrophilic radioiodination technique. This chapter deals also with the medical imaging, techniques of diagnostic nuclear medicine and the purification of radioiodinated compounds using different chromatographic techniques. Since these radioiodinated compounds are used for diagnosis and therapeutic treatment of human diseases, quality control tests such as determination of chemical purity, radionuclidic purity, radiochemical purity, sterility, pyrogenicity and biodistribution are performed to ensure the purity, the safety and efficiency of these products for the intended nuclear medicine application. The third chapter describes the experimental section; comprising chemicals, reagents, the radionuclides, the equipments and the counting systems used in the study. It describes the electrophilic radioiodination using chloramine-T (CAT), iodogen and lactoperoxidase oxidizing agents and the factors affecting the radiochemical yield of the radioiodination of histamine and L-tyrosine methyl ester such as substrate concentration, ph of the medium, reaction time, temperature and stability of the labeled product. This chapter also includes the techniques used in the Purification of radioiodinated compounds, including paper electrophoresis, thin layer chromatography (TLC), Poly acrylamide-acrylic acid resin [P(AAm-AA) resin] and high performance liquid chromatography (HPLC), in addition, the quality control

  1. A radioimmunoassay for erythropoietin: serum levels in normal human subjects and patients with hemopoietic disorders

    Energy Technology Data Exchange (ETDEWEB)

    Rege, A.B.; Brookins, J.; Fisher, J.W.

    1982-12-01

    An RIA for Ep has been developed that is highly sensitive and specific. A homogeneous Ep preparation was labeled with /sup 125/I by the chloramine-T method to a specific activity of 90 to 136 ..mu..Ci/..mu..g and immunoreactivity of 80%. Ep antiserum, which was produced to a human urinary Ep preparation (80 U/mg of protein), was adsorbed with normal human urinary and serum proteins without any loss in sensitivity of the RIA to increase the specificity of the assay. A good correlation was seen between the RIA and the exhypoxic polycythemic mouse assay (corr. coef. 0.967; slope 1.05 and ''y'' intercept 0.75). Ep titers in sera from 175 hematologically normal human subjects exhibited a normal frequency distribution and ranged between 5.8 and 36.6 mU/ml with a mean of 14.9 +/- 4.7 (S.D.) and median of 14.3. Serum Ep titers were markedly elevated in seven patients with aplastic anemia and one patient with pure red cell aplasia (1350 to 20,640 mU/ml) and were lower than normal in two patients with polycythemia vera (8.1 and 9.4 mU/ml). The serum Ep titers in a prenephrectomy patient with chronic glomerulonephritis (31.1 mU/ml) decreased to below normal levels (9.04 mU/ml) after nephrectomy. The cord serum erythropoietin titers in 10 IDM (90.82 +/- 134.1 (S.D.) mu/ml) returned to values within the normal range (13.86 +/- 5.55) on day 3 after birth, suggesting the utility of the RIA in elucidating the role of hypoxia and/or insulin in increased erythropoiesis in IDM. The serum Ep titers in patients with anemias and polycythemias were compared to those of normal human subjects and agreed well with pathophysiologic mechanisms of these hemopoietic disorders, confirming the validity of the RIA.

  2. A radioimmunoassay for erythropoietin: serum levels in normal human subjects and patients with hemopoietic disorders

    Energy Technology Data Exchange (ETDEWEB)

    Rege, A.B.; Brookins, J.; Fisher, J.W.

    1982-12-01

    An RIA for Ep has been developed that is highly sensitive and specific. A homogeneous Ep preparation was labeled with /sup 125/I by the chloramine-T method to a specific activity of 90 to 136 micro Ci/microgram and immunoreactivity of 80%. Ep antiserum, which was produced to a human urinary Ep preparation (80 U/mg of protein), was adsorbed with normal human urinary and serum proteins without any loss in sensitivity of the RIA to increase the specificity of the assay. A good correlation was seen between the RIA and the exhypoxic polycythemic mouse assay (corr. coef. 0.967; slope 1.05 and y intercept 0.75). Ep titers in sera from 175 hematologically normal human subjects exhibited a normal frequency distribution and ranged between 5.8 and 36.6 mU/ml with a mean of 14.9 +/- 4.7 (S.D.) and median of 14.3 Serum Ep titers were markedly elevated in seven patients with aplastic anemia and one patient with pure red cell aplasia (1350 to 20,640 mU/ml) and were lower than normal in two patients with polycythemia vera (8.1 and 9.4 mU/ml). The serum Ep titers in a prenephrectomy patient with chronic glomerulonephritis (32.1 mU/ml) decreased to below normal levels (9.04 mU/ml) after nephrectomy. The cord serum erythropoietin titers in 10 IDM (90.82 +/- 134.1 (S.D.) mu/ml) returned to values within the normal range (13.86 +/- 5.55) on day 3 after birth, suggesting the utility of the RIA in elucidating the role of hypoxia and/or insulin in increased erythropoiesis in IDM. The serum Ep titers in patients with anemias and polycythemias were compared to those of normal human subjects and agreed well with pathophysiologic mechanisms of these hemopoietic disorders, confirming the validity of the RIA.

  3. Functional expression of a single-chain antibody to ErbB-2 in plants and cell-free systems

    Directory of Open Access Journals (Sweden)

    Benevolo Maria

    2006-09-01

    Full Text Available Abstract Background Aberrant signaling by ErbB-2 (HER 2, Neu, a member of the human Epidermal Growth Factor (EGF receptor family, is associated with an aggressive clinical behaviour of carcinomas, particularly breast tumors. Antibodies targeting the ErbB-2 pathway are a preferred therapeutic option for patients with advanced breast cancer, but a worldwide deficit in the manufacturing capacities of mammalian cell bioreactors is foreseen. Methods Herein, we describe a multi-platform approach for the production of recombinant Single chain Fragments of antibody variable regions (ScFvs to ErbB-2 that involves their functional expression in (a bacteria, (b transient as well as stable transgenic tobacco plants, and (c a newly developed cell-free transcription-translation system. Results An ScFv (ScFv800E6 was selected by cloning immunoglobulin sequences from murine hybridomas, and was expressed and fully functional in all the expression platforms, thereby representing the first ScFv to ErbB-2 produced in hosts other than bacteria and yeast. ScFv800E6 was optimized with respect to redox synthesis conditions. Different tags were introduced flanking the ScFv800E6 backbone, with and without spacer arms, including a novel Strep II tag that outperforms conventional streptavidin-based detection systems. ScFv800E6 was resistant to standard chemical radiolabeling procedures (i.e. Chloramine T, displayed a binding ability extremely similar to that of the parental monovalent Fab' fragment, as well as a flow cytometry performance and an equilibrium binding affinity (Ka approximately 2 × 108 M-1 only slightly lower than those of the parental bivalent antibody, suggesting that its binding site is conserved as compared to that of the parental antibody molecule. ScFv800E6 was found to be compatible with routine reagents for immunohistochemical staining. Conclusion ScFv800E6 is a useful reagent for in vitro biochemical and immunodiagnostic applications in oncology

  4. Improved iodine radiolabels for monoclonal antibody therapy.

    Science.gov (United States)

    Stein, Rhona; Govindan, Serengulam V; Mattes, M Jules; Chen, Susan; Reed, Linda; Newsome, Guy; McBride, Bill J; Griffiths, Gary L; Hansen, Hans J; Goldenberg, David M

    2003-01-01

    A major disadvantage of (131)iodine (I)-labeled monoclonal antibodies (MAbs) for radioimmunotherapy has been the rapid diffusion of iodotyrosine from target cells after internalization and catabolism of the radioiodinated MAbs. We recently reported that a radioiodinated, diethylenetriaminepentaacetic acid-appended peptide, designated immunomedics' residualizing peptide 1 (IMP-R1), was a residualizing iodine label that overcame many of the limitations that had impeded the development of residualizing iodine for clinical use. To determine the factors governing the therapeutic index of the labeled MAb, as well as the factors required for production of radioiodinated MAb in high yield and with high specific activity, variations in the peptide structure of IMP-R1 were evaluated. A series of radioiodinated, diethylenetriaminepentaacetic acid-appended peptide moieties (IMP-R1 through IMP-R8) that differed in overall hydrophilicity and charge were compared. Radioiodinations of the peptides followed by conjugations to disulfide-reduced RS7 (an anti-epithelial glycoprotein-1 MAb) furnished radioimmunoconjugates in good overall incorporations, with immunoreactivities comparable to that of directly radioiodinated RS7. Specific activities of up to 8 mCi/mg and yields > 80% have been achieved. In vitro processing experiments showed marked increases in radioiodine retention with all of the adducts; radioiodine retention at 45 h was up to 86% greater in cells than with directly iodinated RS7. Each of the (125)I-peptide-RS7 conjugates was compared with (131)I-RS7 (labeled by the chloramine-T method) in paired-label biodistribution studies in nude mice bearing human lung tumor xenografts. All of the residualizing substrates exhibited significantly enhanced retention in tumor in comparison to directly radioiodinated RS7, but the nontarget uptakes differed significantly among the residualizing labels. The best labels were IMP-R4 and IMP-R8, showing superior tumor-to-non-tumor ratios

  5. Distribution of {sup 131} I- labeled Bothrops erythromelas venom in mice

    Energy Technology Data Exchange (ETDEWEB)

    Vasconcelos, C.M.L.; Valenca, R.C.; Araujo, E.A. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia; Modesto, J.C.A.; Pontes, M.M.; Guarnieri, M.C. [Pernambuco Univ., Recife, PE (Brazil). Dept. de Zoologia; Brazil, T.K. [Bahia Univ., Salvador, BA (Brazil). Inst. de Biologia

    1998-03-01

    Bothrops erythromelas is responsible for many snake bites in northeastern Brazil. In the present study we determined the in vivo distribution of the venom following its subcutaneous injection into mice. B. erythromelas venom and albumin were labeled individually with {sup 131} I by the chloramine T method, and separated in a Sephacryl S-200 column. The efficiency of labeling was 68%.Male Swiss mice (40-45 g), which had been provided with drinking water containing 0.05% KI over a period of 10 days prior to the experiment, were inoculated dorsally (sc) with 0.3 ml (2.35 x 10{sup 5} cpm/mouse) of {sup 131} I-venom (N = 42), {sup 131} -albumin or {sup 131} I (controls, N = 28 each). Thirty minutes and 1,3, 6, 12, 18 and 24 h after inoculation, the animals were perfused with 0.85% Na Cl and skin and various organs were collected in order to determine radioactivity content. There was a high rate of venom absorption int he skin (51%) within the first 30 min compared to albumin (20.1%) and free iodine (8.2%). Up to the third hour after injection there was a tendency for venom and albumin to concentrate in the stomach ( 3 rd h),small intestine (3 rd h) and large intestine (6th h). Both control groups had more radioactivity in the digestive tract, especially in the stomach, but these levels decreased essentially to baseline by 12-18 h postinjection. In the kidneys, the distribution profiles of venom, albumin and iodine were similar. Counts at 30 min postinjection were low in all three groups (1.37, 1.86 and 0.77, respectively), and diminished to essentially 0% by 12-18 h. Albumin tended to concentrate in muscle until the 3 rd h postinjection (1.98%).There was a low binding of labeled venom in the liver (B. erythromelas venom does not specifically target most internal organs). That is, the systemic effects of envenomation ar mainly due to an indirect action. (author) 14 refs., 2 figs.

  6. Monoclonal antibody RM2 as a potential ligand for a new immunotracer for prostate cancer imaging

    International Nuclear Information System (INIS)

    Objectives: To investigate the potential of monoclonal antibody (mAb) RM2 as a ligand for a radioimmunotracer for prostate cancer imaging. Methods: Labeling was conducted with mAb RM2 and 125I using the chloramine-T method. The cell study was conducted with PC-3 and LNCaP, which are prostate cancer cell lines, and MCF-7, which is a breast cancer cell line. The cells were treated or untreated with unlabeled mAb RM2 to block the haptoglobin-β chains expressed on the surface of the prostate cancer cells. 125I-mAb RM2 was added into the cell culture media and cellular uptake of 125I-mAb RM2 was evaluated at 1, 3 and 6 hours of incubation. For the in vivo biodistribution study, PC-3 cells were implanted in athymic male mice. The animals were injected intravenously with 125I-mAb RM2. At 24, 48 and 72 hours after tracer injection, the animals were sacrificed and the activity levels of blood and tissue samples were determined. Results: The uptake of 125I-mAb RM2 in the PC-3 and LNCaP cells increased according to the incubation time, while the uptake of 125I-mAb RM2 in MCF-7 cells did not show any increase up to 6 hours. The increase of 125I-RM2 uptake was not observed when the PC-3 and LNCaP cells were pre-treated with unlabeled RM2. In the biodistribution studies, 125I-mAb RM2 showed marked uptake into the implanted PC-3 cells. In PC-3 tumor-bearing mice, the tumor muscle ratio of 125I-RM2 was increased for up to 72 hours in a time-dependent manner. Conclusions: 125I-mAb RM2 showed excellent prostate cancer cell targeting in vitro and in vivo. Therefore, mAb RM2 seems to be a potential candidate for an immunoligand for prostate cancer imaging.

  7. Synthesis and evaluation of iodine-123 labelled tricyclic tropanes as radioligands for the serotonin transporter

    Energy Technology Data Exchange (ETDEWEB)

    Quinlivan, Mitchell; Mattner, Filomena; Papazian, Vahan; Zhou, Jia; Katsifis, Andrew; Emond, Patrick; Chalon, Sylvie; Kozikowski, Alan; Guilloteau, Denis; Kassiou, Michael E-mail: mkassiou@med.usyd.edu.au

    2003-10-01

    The tricyclic tropane analogues (1S,3S,6R,10S)-(Z)-10-(benzoyloxymethyl)-9-(3-chloro-4-iodobenzylidene)-7 -azatricyclo[4.3.1.0{sup 3,7}]decane, 1, and (1S,3S,6R,10S)-(Z)-9-(3-chloro-4-iodobenzylidene)-7-azatricyclo[4.3.1.0{sup 3,7}] = decane-10-carboxylic acid methyl ester, 2, have been shown to be potent and selective serotonin transporter (SERT) ligands. They possess nanomolar affinity for the SERT (Ki = 0.06 nM and 1.8 nM respectively) and are suitable for radiolabelling using iodine-123. In the present study we prepared [{sup 123}I]1 and [{sup 123}I]2 from the appropriate tributylstannane precursors using acidic media with chloramine-T as the oxidising agent. The radiochemical yield obtained for [{sup 123}I]1 varied between 50-60% while for [{sup 123}I]2 the range was 65-80%. Both radioligands were obtained with radiochemical purity > 97% and specific activity estimated to be > 185 GBq/{mu}mol. The biodistribution of [{sup 123}I]1 demonstrated low degree of brain penetration at 5 min (0.14%ID/g) with a homogenous distribution. The radioactivity cleared quickly from all brain regions with no preferential localization. In comparison, [{sup 123}I]2 demonstrated on average a higher brain uptake at 5 min (0.5%ID/g). However the distribution of radioactivity was homogenous and cleared to levels similar to [{sup 123}I]1 at 1 hr post-injection. Pre-administration of citalopram failed to show any significant inhibition of [{sup 123}I]2 uptake in the rat brain. The high lipophilicity of 1 and 2 (HPLC-derived log P{sub 7.4} values of 6.41 and 4.25 respectively) and in vivo metabolism, seen by high thyroid uptake would explain the absence of any specific binding observed in the rat brain. In view of these results [{sup 123}I]1 and [{sup 123}I]2 do not appear to be suitable radioligands for in vivo studies of the SERT.

  8. Experimental study of radiotargeting-therapy with small molecular polypeptide in nude mice bearing lung adenocarcinoma

    International Nuclear Information System (INIS)

    Background: Integrin signal transduction pathways provide an important basis for molecular targeting therapy of cancer in tumor growth, infiltration and transfer. Existing research data have shown that small molecular peptide labeled with radionuclide has good clinical application prospects, but the successful researches on lung cancer have not been reported so far. It is considered that the main reason is the lack of small molecule peptide for specific targeting lung cancer. Purpose: Based on the small molecular peptide cNGQGEQc for specifically identifying integrin α3 and β1 found previously, polypeptide cNGQGEQc is selected and radiolabelled with 131I. And the inhibitory effect of 131I-cNGQGEQc in nude mice bearing lung adenocarcinoma is observed. Methods: The coupling of cNGQGEQc and tyrosine was done in the processing of solid phase synthesis of small molecular peptide. Chloramine-T method was used for radiolabelling of cNGQGEQc with 131I. Twenty nude mice bearing NCI-H1975 were built and randomly divided into four groups with five mice in each group, including the group of 131I-cNGQGEQc, the group of 131I-cNAQAEQc, the group of 131I and the saline control group. The general condition was observed in nude mice bearing tumor after tail vein injection of corresponding drugs. And the tumor sizes after grafting were measured per 3 days in 30 days. The inhibitory rate of tumor in each group was calculated. Results: The labeling efficiencies of 131I-cNGQGEQc and 131I-cNAQAEQc were greater than 90% with the radiochemical purity of more than 95%, and 131I-cNGQGEQc had obvious inhibitory effect for transplantation tumor in nude mice bearing NCI-H1975 adenocarcinoma of lung. After a treatment for 30 days the tumor inhibitory rates were 60.93% for the group of 131I-cNGQGEQc, 11.63% for the group of 131I-cNAQAEQ and 10.70% for the group of 131I. Conclusion: 131I-cNGQGEQC has a good affinity and effective inhibit effect for the NCI-H1975 lung adenocarcinoma. Integrin is

  9. Preparation of the radiopharmaceutical {sup 131}I-Anti-CD20 for the treatment of lymphomas; Preparacion del radiofarmaco {sup 131}I-Anti-CD20 para el tratamiento de linfomas

    Energy Technology Data Exchange (ETDEWEB)

    Pantoja H, I.E

    2004-07-01

    At the present time they are considered to the lymphomas like a problem of first magnitude since has happened it is necessary to be the fifth cancer cause in the world. Different treatments focused to the lymphoma like the chemotherapy and the radiotherapy, have been employees to counteract the No-Hodgkin lymphoma, without these they don't exclude the healthy tissue of the toxicity. It is for it that is taking a new direction with the employment of the directed radioimmunotherapy since this it allows to kill wicked cells selectively with radiation dose joined to the apoptosis and cytotoxicity induced by the own one bio molecule. The radioimmunotherapy with radiolabelled antibodies directed to the surface antigen CD20 represents a new modality for the treatment of No-Hodgkin lymphoma and potentially other illnesses. In this work the parameters of optimization are presented for the preparation, control of quality and evaluation of the stability in vitro and in vivo of the monoclonal antibody anti-CD20 labelled with {sup 131} I for the treatment of No-Hodgkin lymphoma. The anti-CD20 labelled by the chloramine-T method with high radiochemical purity (>98%), it is stable in solution for but of a half life of the radionuclide (8.04 days) The {sup 131} I-anti-CD20 doesn't present dehalogenation in vitro (human serum) during 24 h of incubation at 37 C. According to the tests carried out to establish the immunoreactivity, a percentage of union to cells was obtained (B lymphocytes) bigger to 30%. The biodistribution in mice balb/c one hour after their administration, it shows that there is not high reception in mucous neither kidneys, what indicates that the complex is stable in vivo. In conclusion, the radiopharmaceutical {sup 131} I-anti-CD20 was obtained in sterile injectable solution and free of pyrogens with a radiochemical purity bigger to 98% and a specific activity of 296 MBq. The radiolabelled molecule maintains its biological recognition for the receiving

  10. Preparation of the radiopharmaceutical 131I-Anti-CD20 for the treatment of lymphomas

    International Nuclear Information System (INIS)

    At the present time they are considered to the lymphomas like a problem of first magnitude since has happened it is necessary to be the fifth cancer cause in the world. Different treatments focused to the lymphoma like the chemotherapy and the radiotherapy, have been employees to counteract the No-Hodgkin lymphoma, without these they don't exclude the healthy tissue of the toxicity. It is for it that is taking a new direction with the employment of the directed radioimmunotherapy since this it allows to kill wicked cells selectively with radiation dose joined to the apoptosis and cytotoxicity induced by the own one bio molecule. The radioimmunotherapy with radiolabelled antibodies directed to the surface antigen CD20 represents a new modality for the treatment of No-Hodgkin lymphoma and potentially other illnesses. In this work the parameters of optimization are presented for the preparation, control of quality and evaluation of the stability in vitro and in vivo of the monoclonal antibody anti-CD20 labelled with 131 I for the treatment of No-Hodgkin lymphoma. The anti-CD20 labelled by the chloramine-T method with high radiochemical purity (>98%), it is stable in solution for but of a half life of the radionuclide (8.04 days) The 131 I-anti-CD20 doesn't present dehalogenation in vitro (human serum) during 24 h of incubation at 37 C. According to the tests carried out to establish the immunoreactivity, a percentage of union to cells was obtained (B lymphocytes) bigger to 30%. The biodistribution in mice balb/c one hour after their administration, it shows that there is not high reception in mucous neither kidneys, what indicates that the complex is stable in vivo. In conclusion, the radiopharmaceutical 131 I-anti-CD20 was obtained in sterile injectable solution and free of pyrogens with a radiochemical purity bigger to 98% and a specific activity of 296 MBq. The radiolabelled molecule maintains its biological recognition for the receiving CD20 highly expressed in

  11. Iodine labelled diethylstilbestrol (DES) of high specific activity: A potential radiopharmaceutical for therapy of estrogen receptorpositive tumours and their metastases?

    International Nuclear Information System (INIS)

    After binding to the estrogen receptor (ER) estrogens are localized very close to the DNA. Therefore, radioactively labelled estrogens, especially in connection with Auger-electron emitters which have only a very short range (1-10 nm) of radiation, are excellent candidates to achieve high specific cytotoxicity, in combination with a low degree of side effects. Diethylstilbestrol (3,4-Di-(4'-hydroxyphenyl)-hexene-3) is a well-known, synthetic, non-steroidal estrogen, which has a higher affinity to the estrogen receptor than the natural hormone estradiol itself. The idea to use iodine labelled DES for imaging of ER positive tumours is not new. Several working groups successfully tried to label DES with different methods and investigated the diagnostic usefulness of the product. Most attempts for labelling with radioiodine have been made with a water soluble derivate of DES, the tetra-sodium salt of diethylstilbestrol-diphosphate (DES-2P). Former labelling methods had some disadvantages: low yield (20-30%), low specific activity (0.7 - 2.9 GBq/mmol), bad reproducibility, and time consuming procedures. Presumably, the low specific activity was the reason for the unsatisfying biodistribution data observed with this labelled compounds. In the present work a fast and simple labelling and purification method for *I-DES was used and its binding affinity for ER was determined. For testing its cytotoxic effects on MCF-7 mamma carcinoma cells different iodine isotopes bound to DES or in form of iodide were compared with regard to apoptosis, necrosis, and viability. Last but not least first animal experiments with tumour bearing mice were carried out. DES was iodinated by Chloramine T in methanolic solution. Purification and quality control was carried out with reversed phase HPLC (column: Column: Hypersil ODS, 250 x 4 mm, 10 Ym, Eluent A: Methanol G, Eluent B: Water G, Gradient: 20% A to 70% A within 5 min, Flow: 1 ml/min, UV-Detection: 254 nm). The dissociation constants for

  12. Comparison of [131I]-Tyr3-octreotate and [131I]DOTA-Tyr3-octreotate: the effect of DOTA on the pharmacokinetics and stability

    International Nuclear Information System (INIS)

    The introduction of radiolabeled somatostatin analogs for peptide receptor imaging and therapy of neuroendocrine cancer have become a primary focus of interest in nuclear medicine. In this work we studied the possibility of preparing radioiodinated octreotate derivatives, with high stability and favourable kinetic in vivo, because 131I-iodide is most frequently used in therapeutic nuclear medicine and aviable. We studied molar peptide to radionuclide ratio in order to obtain the mono-iodinated peptide (di-iodinated no longer binds to the somatostatin receptor). Like other radioiodinated proteins labeled on constituent tyrosine residues, it was important to study the possibility of dehalogenation in vivo. Despite DOTA chelating group was not necessary to the radioiodination procedure, we intended to evaluate the influence of the chelating group on the pharmacokinetic and the in vivo stability of the labeled peptide. 131I radiolabeling of Tyr3-octreotate and DOTA-Tyr3-octreotate was performed using Chloramine T method. A solution of 10 μg of peptide/40 μL of PBS (0.05M phosphate-buffered saline pH 7.5) was transferred to an Eppendorf cap. After addition of 10 μL (74 MBq) radioiodine and 5 μL of chloramine T solution (1mg/mL PBS), the cap was carefully stirred and the labeling reaction was allowed to proceed for 3 minutes at room temperature; 5 μL of sodium methabisulfite solution (2mg/mL PBS) was introduced as reducing agent. Radiochemical purity was determined by HPLC (Waters) using a RP C18 column (4.2 x 50 mm, 5 μm, Waters) with UV (230 nm) and radioactivity (Packard Canberra) detection, flow rate of 0.5 mL/minute with a linear gradient of 40-80% (v/v) methanol in 50 mM sodium acetate buffer (pH 5.5) for 20 minutes, maintained for another 25 minutes. Free radioiodine was also determined by horizontal zone electrophoresis (Amershan Pharmacia) on Whatman 1 paper, 0.05M barbital buffer, pH 8.6, 300V, 40 minutes. The stability of the compounds were evaluated

  13. Utilization of N-X bonds in the synthesis of N-heterocycles.

    Science.gov (United States)

    Minakata, Satoshi

    2009-08-18

    Nitrogen-containing heterocycles--such as aziridines, pyrrolidines, piperidines, and oxazolines--frequently show up as substructures in natural products. In addition, some of these species show potent biological activities. Therefore, researchers would like to develop practical and convenient methods for constructing these heterocycles. Among the available methods, the transfer of N(1) units to organic molecules, especially olefins, is a versatile method for the synthesis of N-heterocycles. This Account reviews some of our recent work on the synthesis of N-heterocycles using the N-X bond. A nitrogen-halogen bond bearing an electron-withdrawing group on the nitrogen can be converted to a halonium ion. In the presence of C-C double bonds, these species produce three-membered cyclic halonium intermediates, which can be strong electrophiles and can produce stereocontrolled products. N-Halosuccinimides are representative sources of halonium ions, and the nitrogen of succinimide is rarely used in organic synthesis. If the nitrogen could act as a nucleophile, after releasing halonium ions to C-C double bonds, we expect great advances would be possible in the stereoselective functionalization of olefins. We chose N-chloro-N-sodio-p-toluenesulfonamide (chloramine-T, CT), an inexpensive and commercially available reagent, as our desired reactant. In the presence of a catalytic amount of CuCl or I(2) and AgNO(3), we achieved the direct aziridination of olefins with CT. The reaction catalyzed by I(2) could be carried out in water or silica-water as a green process. The reaction of iodoolefins with CT gave pyrrolidine derivatives under extremely mild conditions with complete stereoselectivity. We also extended the utility of the N-chloro-N-metallo reagent, which is often unstable and difficult to work with. Although CT does not react with electron-deficient olefins without a metal catalyst or an additive, we found that N-chloro-N-sodiocarbamates react with electron

  14. Comparative biodistribution profile of [131I]VIP and [131I]VIP10-28

    Directory of Open Access Journals (Sweden)

    Maria Tereza Colturato

    2005-10-01

    Full Text Available Various tumor cells express significantly higher amounts of VIP receptors (VIPR that provided the basis for the clinical use of radiolabeled VIP for the in vivo localization of tumors. This work studied the labeling of VIP and VIP10-28 with iodine-131 to compare the biological distribution of the labeled compounds in Nuce mice and the affinity for tumor cells. Both VIP and VIP10-28 peptides contain two tyrosine residues, in positions 10 and 22, that are theoretically equally susceptible to radioiodination employing oxidative electrophilic substitution using oxidizing agents like Chloramine T. Radiochemical purity of the reaction mixture was determined by electrophoresis and HPLC. The VIP peptide and the fragment were labeled with radioiodine with good radiochemical yield (above 96%. Suitable, but important differences can be observed in biological distribution studies. Comparatively, blood clearance was faster for labeled VIP and perhaps because of this, the uptake in tumor was lower, especially during the first hour. These differences observed in the biological distribution of the compounds can be related to the lipophilicity of the labeled compounds.Várias células tumorais expressam significantemente uma alta quantidade de receptores VIP (VIPR que determinam a base para o uso clínico de VIP radiomarcado para localização de tumores in vivo. Foi estudado neste trabalho a marcação do VIP e do fragmento VIP10-28 com iodo-131 comparando a distribuição biológica dos compostos marcados em camundongos Nude e sua afinidade pelas células tumorais. Ambos os peptídeos, VIP e VIP10-28. contém dois resíduos de tirosina nas posições 10 e 22, que teoricamente são igualmente susceptíveis pela substituição eletrofílica oxidativa do radioiodo utilizando Cloramina T como agente oxidante. A pureza radioquímica da mistura de reação foi determinada por eletroforese e cromatografia líquida de alta eficiência (CLAE. O VIP e fragmento foram

  15. SYSTEMATIC EVALUATION ON DISINFECTION EFFECICACY OF DIFFERENT CHLORINE DISINFECTANTS%不同种类含氯消毒剂消毒效果研究的系统评价

    Institute of Scientific and Technical Information of China (English)

    刘真; 张林; 熊鸿燕

    2011-01-01

    Disinfection; 37 English literatures are distributed in 17 journals, and 37.84% are published in Water Research, 13.51% are published in Journal of Hospital Infection.Chlorine, chlorine dioxide, sodium hypochlorite, sodium dichloroisocyanurate, trichloroisocyanuric acid ( TCCA), calcium hypochlorite and chloramine - T are involved in the 102 literatures.The order of comprehensive sterilization effect is as follows (from strong to weak): chlorine dioxide, TCCA, sodium dichloroisocyanurate, sodium hypochlorite.Coclusion Numbers of studies on bactericidal effect of chlorine disinfectants still leap into the front ranks among the studies of chemical disinfectant in the first decade of this century.Chlorine dioxide, sodium hypochlorite, sodium dichloroisocyanurate, and trichloroisocyanuric acid are the most widely used and studied.

  16. Labelling and biological valuation of anti-CD-20 for treatment of non-Hodgkin's lymphoma

    International Nuclear Information System (INIS)

    Full text: Anti-CD20 monoclonal chimeric humanized murine antibodies (Rituximab), have been successfully applied for the treatment of Non Hodgkin's Lymphoma. However, upon labelling of the mab-CD20 with β-emitters as 90Y, the therapeutic efficacy has significantly increased due to radiological effects of ionizing radiation. Our objective was to develop reliable and efficient methods for labelling anti-CD20 with β-emitters of therapeutic interest and simple and rugged quality control methods to evaluate radiochemical purity, biological performance and immunoreactivity assessment. 131I and 188Re have been used for the labelling of anti-CD20 as two attractive alternatives due to decay properties and availability (188Re: Eβmax: 2.2MeV, Eγ 0,155MeV, T=17h, generator produced; 131I: Eβmax: 0,63MeV, Eγ 0,364MeV, T=8d). Labelling of anti-CD20 was optimized following the oxidation procedure of chloramine-T in the case of 131I and the synthesis of 188Re(IV) complex with the previously reduced monoclonal antibody. Quality control of the species obtained were done by physicochemical methods, including ITLC-SG and HPLC, non specific protein precipitation, biological distribution in normal mice and immunoreactivity studies with membrane antigens extracted from isolated leucocytes. 131I- (more than 3.7 GBq/mL) was introduced on tyrosyl residues of the protein chain by adding 28 MBq to 20μg of anti CD20 (Mab Thera, 10mg/mL) at pH 7.4 and 1.3μg of Choramine-T. Purification was done by gel-permeation with sephadex G-25 (PD-10, Pharmacia). For labelling with 188Re, anti CD20 was first reduced by incubation with 2- mercaptoethanol and purified over a PD10 column. Fractions of reduced antibody were pooled and formulated as kit for instant labelling. Each kit contained 1mg anti-CD20; 82.8mg of sodium tartrate; 1.67 mg of stannous fluoride and 0.25 mg gentisic acid. For the labelling, sodium perrhenate (1.5-1.9 GBq) was acidified, added to the kit and then incubated for 1 hour at

  17. Study on biodistribution of 131 iodine labeled monoclonal antibody D-D3 against pro-gastrin-releasing peptide(31-98) in healthy Kunming mice%抗ProGRP(31-98)单克隆抗体D-D3的131I标记及其体内生物学分布研究

    Institute of Scientific and Technical Information of China (English)

    陈传新; 石怡珍; 杨仪; 唐军; 刘增礼; 徐巧玲

    2011-01-01

    Objective To study the 131I labeling methods, stability, and biological distribution pattern of D-D3 antibody against pro-gastrin-releasing peptide 31-98(ProGRP(31-98) ). Methods The radioiodination of D-D3 antibody was performed using the chloramine-T method. The radiochemical purity was determined through thin-layer chromotography. 131I-D-D3 was injected into the healthy Kunming mice via a tail vein, and the % ID/g for various organs was obtained, and then, the biodistribution and pharmacokinetics of 131 I-D-D3 antibody in healthy Kunming mice were studied. Results The 131 I-D-D3 labeling rate was (86.56 ± 3.8) %. The radiochemical purity of 131 I-D-D3 was (99.27 ± 0. 6)%. After 48 h incubating in 37 ℃ water bath, the radiochemical purity was (88.38 ± 0.4)%. While being mixed 24 h with healthy human serum, the radiochemical purity was still more than (64.43 ± 0.7)%. The metabolism of 131I-D-D3 in healthy Kunming mice was consistent with a two-compartment model with first-order absorption, T1/2α and T1/2β was 0.25,37.89 h, respectively. Conclusion The labeling efficiency and radiochemical purity of 131 I-D-D-D3 are high and stable. 131I-D-D3 is a promising radioimmunoimaging reagent for small cell lung cancer(SCLC).%目的 探讨抗胃泌素释放肽前体(ProGRP)(31-98)单克隆抗体D-D3的131I标记方法 及其在健康昆明小鼠体内的生物学分布规律与特点.方法 采用氯胺-T法用131I标记单克隆抗体D-D3,利用纸层析法测定其标记率、放化纯度和稳定性.取健康昆明小鼠50只,随机分为10组,每组5只,自昆明小鼠尾静脉注射131I-D-D3 1.48 kBq/100 μL(4 μCi/100 μL),各组小鼠分别于注射后5、15、30 min及1、2、4、8、12、24、48 h处死,取血液、心脏、肝脏、脾脏、肺脏、肾脏、胃、小肠、骨骼(右下肢)、肌肉(右下肢)和脑组织,称质量(g)后测放射性计数(cpm),计算各脏器组织的每克组织百分注射剂量率(%ID

  18. Effect of cardiac muscle collagen and actin on myocardiopathological change of diabetes mellitus%心肌胶原蛋白和肌动蛋白对糖尿病心肌病变的研究

    Institute of Scientific and Technical Information of China (English)

    吴伟; 张锦; 邱阳

    2005-01-01

    目的研究糖尿病大鼠不同病程心肌胶原蛋白和骨架蛋白含量的变化,阐明两者对糖尿病心肌病变发生的作用.方法制造糖尿病大鼠心肌模型随机分组.氯胺T法测定羟脯氨酸含量,代表心肌胶原总含量.心肌免疫组织化学染色测定心肌胶原蛋白(Collagen Ⅰ、Collagen Ⅲ)和心肌型α肌动蛋白(α-actin)及转化生长因子β1(TGF-β1)平均积分光密度(IOD).心肌病理改变的光镜和透射电镜观察.结果糖尿病痛程6个月组心肌胶原总含量明显高于病程3个月以内组(P<0.01).病程3个月之后Ⅰ型胶原蛋白表达伴随TGF-β1的表达开始较健康鼠明显增加(P<0.01).α-actin蛋白表达较健康鼠明显减少(P<O.01).糖尿病鼠心肌横切面可见粗大胶原纤维相互连接成网状,排列紊乱,分布不匀.心肌细胞心肌型α-actin蛋白分布不均匀,着色呈浅黄色,心肌纵切面可见α-actin蛋白表达主要分布于心肌肌膜处.病程3个月后大鼠心肌细胞核皱缩,线粒体肿胀、模糊,闰盘不连续,α-actin蛋白表达明显减少,有糖原沉积现象.结论Ⅰ型心肌胶原蛋白呈现持续性增加是糖尿病鼠心肌纤维化的主要原因.心肌细胞核皱缩,线粒体肿胀、模糊,闰盘不连续,糖原沉积和心肌型actin表达减少是糖尿病心肌病病理基础.%[Objective] To investigate the content changes of myocardial collagenprotein and actin at different stages of diabetes mellitus(DM) rats. So as to illustrate that they play a role in myocardial changes of DM. [Methods] Cardiac muscle model of DM rats was grouped randomly. The hydroxy proline contents were measured with the chloramine T method. The type Ⅰ collagen, type Ⅲ collagen,transforming factor beta1 (TGF-beta1) and cardiactype α-actin were determined by myocardial immunehistochemical stain. Myocardiopathological change were observed with light microscope and transmission electron microscope. [Results] The collagen contents

  19. Preclinical study of radioiodinated glucose-Tyr3-octreotate: Comparison with 111In-DOTA-Tyr3-octreotate

    International Nuclear Information System (INIS)

    Full text: Targeted radionuclide imaging and treatment are based upon the interaction of radioligands with receptors in the target tissue (namely high density receptor specific tumours). As receptors for somatostatin (mainly somatostatin receptor subtype-2 /sstr2/) are over-expressed in several human tumours of endocrine origin, a number of radiolabeled somatostatin analogs have been recently introduced as the vectors for targeted imaging and therapy; commercially available 111In-DTPA-octreotide being a gold standard in this field. Several publications demonstrate that a substitution of Tyr instead of Phe in the peptide position 3 and oxidation of carboxyl end of octreotide significantly increased the affinity of the peptide to sstr2. More recently it has been shown that NH2-terminal carbohydration leads to a further improvement of the peptide receptor affinity and its retention in the tumour. In this study we present preclinical analysis of distribution profile and elimination pathways of radioiodinated glucose-Tyr3-octreotate (125I-gluc-TOCA) in comparison with that of another promising targeted radiopharmaceutical, namely 111In-DOTA-Tyr3-octreotate (111In- DOTA-TOCA). Gluc-TOCA was radioiodinated using chloramine-T method. For radiolabeling of DOTA-TOCA with radiometal, to 200 μl of 0.4 M acetate buffer with 0.24 M gentisic acid pH 5, 10μl of peptide solution in 0.1% TFA (1mg/ml) were added together with 0.5-1 mCi of 111InCl3 in 0.04 M HCl. Reaction mixture was heated 25 minutes to 90-95 deg. C. Pharmacokinetic studies were performed on male Wistar rats. Results confirmed that specific internalization of 125I-gluc-TOCA by sstr2 - expressing AR423 rat pancreatic carcinoma cells in vitro was about twice of that for 111In-DOTA-TOCA. In rats, 125I-gluc-TOCA exhibited prolonged plasma clearance in comparison with the other peptide. Slower decrease of plasma radioactivity after 125I-gluc-TOCA was due to its higher lipophilicity and thus higher plasma protein binding

  20. Effect of intestinal ischemia-reperfusion injury on the leptin levels in serum and adipose tissue%肠缺血再灌注损伤对血清及脂肪组织Leptin水平的影响

    Institute of Scientific and Technical Information of China (English)

    林季; 颜光涛; 王录焕; 郝秀华; 张凯; 薛辉

    2004-01-01

    BACKGROUND: Recent studies suggest that adipose tissue is not only the reservation of fat, but also a multi-potential endocrine organ secreting many functional cytokines. Leptin is a protein specifically secreted by adipose tissue, and primarily reduces food-intake and promotes energy expenditure.OBJECTIVE: To explore the effect of intestinal ischemia-reperfusion injury on leptin levels in serum and adipose tissue, and find out the role of leptin playing in acute inflammatory responses.DESIGN: A completely random self-and mutual-control experiment.SETTING and MATERIALS: The experiment was collected and completed in Research Laboratory of Biochemistry, General Hospital of Chinese PLA.Three male New Zealand white rabbits and fifty-four male Sprague-Dawley rats were used for experimental use.INTERVENTIONS: New Zealand white rabbits were immunized to obtain anti-leptin, leptin antigen was iodinated by chloramines-T method, thus we established a concise radioimmunoassay for murine leptin. An intestinal ischemia-reperfusion injury model of rats was established, and rats were divided randomly into six groups: sham-operation group(sham), 60-minute ischemia and 30-minute reperfusion group(I60-R30), I60-R90, I60-R150,I60-R240 and I60-R360; each group contained nine rats.MAIN OUTCOME MEASURES: The changes of leptin concentrations in serum and adipose tissue before and after ischemia-reperfusion injury were checked by the radioimmunoassay.RESULTS: Compared with self-control(before injury), serum leptin level of I60-R30 decreased significantly(t=2.3891, P<0.05), that of I60-R150 expressed a trend to increase and that of I60-R360 increased significantly ( t = - 2. 3437, P<0.05 ). Compared with sham after injury [ (9.88±1.87)μg/L], serum leptin level of I60-R240 expressed a trend to increase, that of I60-R360[ (19.43±2.84) μg/L] increased significantly( t = - 2. 8085, P <0.05) . Compared with sham after injury [ ( 11.12±1.27 ) ng], adipose leptin levels of I60-R30[ (4

  1. 益气养阴、活血化瘀中药对糖尿病大鼠脂代谢紊乱及蛋白质非酶糖基化的影响%Effect of Chinese herb for benefiting qi, nourishing yin, activating blood circulation and resolving stasis on lipid metabolic disturbance and protein nonenzyme glycosylation in diabetic rats

    Institute of Scientific and Technical Information of China (English)

    戴红

    2005-01-01

    administered once every day in above 6 groups,and the treating course was 8 weeks. ③ Before treatment, on the 3rd, 6th and 8th weeks after treatment, fasting blood sugar was tested with One Touch Ⅱ tester. 8 weeks later, automatic biochemistry analyzer was used to assay triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDLC) and high-density lipoprotein cholesterol (HDLC). Thoracic aorta was collected swiftly and chloramine-T method was used to assay hydroxyproline content in the extract from aorta. Hydroxyproline content ×7.14=collagen content. Fluorescence spectrometer was used to assay the fluorescence degree of AGEs in collagen extract. ④ t test was applied for difference comparison of measurement data.MAIN OUTCOME MEASURES: ① Comparison of fasting blood sugar before and after treatment in each group. ② Comparison of blood lipid,collagen content and fluorescence degree of collagen-protein AGEs in thoracic aorta in 8 weeks after treatment in each group.RESULTS: At the end of experiment, 8, 4, 2, 2 and 3 rats were died in model group, insulin group, HXY low, middle and high-dose groups respectively, due to which, 12, 8, 11, 13, 13 and 12 rats entered result analysis in various groups respectively. ① Fasting blood sugar: Before treatment, it was higher remarkably in model and various treating groups compared with normal control (P < 0.01) and 8 weeks after treatment, it was lower remarkably in insulin group and various HXY groups compared with model group and before treatment (P < 0.01). ② Levels of TC, TG, LDLC in serum: Those in insulin and model groups were higher remarkably than normal control (P < 0.05-0.01). Those in insulin group and various HXY groups were lower remarkably than normal group (P < 0.05-0.01). ③ Level of HDLC: It was lower remarkably in insulin group and model group compared with normal control (P < 0.05-0.01), it was higher remarkably in HXY low and middle-dose groups compared with model group (P