Sample records for chloramine-t

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

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


    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).

  2. Survival of cool and warm freshwater fish following chloramine-T exposure (United States)

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


    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.

  3. 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)


    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.)

  4. Thermodynamics of Micellization of Nonionic Surfactant Tween-40 in Presence of Additive Chloramine-T Using Clouding Phenomenon

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    A. A. Patil


    Full Text Available The phenomenon of solubilization of nonionic surfactant Tween-40 has been studied through the influence of additive chloramine-T in aqueous medium by measuring the cloud points (CP of the pure surfactant and with chloramine-T. The CP of pure surfactant was found to be increased with increasing concentration of Tween-40. The CP of mixed system shows increasing trends with increased chloramine-T. This is mainly due to increased micelle concentrations. The influence of chloramine-T on the cloud point of Tween-40 is a clear indication that the phenomenon of clouding is associated with the different micelles coalescing. Considering cloud point as threshold temperature of the solubility, the thermodynamic parameters of clouding process (ΔG0cl, ΔH0cl and ΔS0cl have been evaluated using “Phase Separation Model”. The phase separation results from micelle-micelle interaction. It was found that the overall clouding process was exothermic and ΔH0cl > TΔS0cl indicating that the process of clouding was guided by both enthalpy and entropy. This work supports the conjecture that the clouding is critical phenomenon rather than the growth of micelles. Findings of the present work supports to made the probable evidence of electrolyte-surfactant interactions in aqueous medium.

  5. Catalyst-Free Imidation of Allyl Sulfides with Chloramine-T and Subsequent [2,3]-Sigmatropic Rearrangement

    Institute of Scientific and Technical Information of China (English)

    江玉波; 莫凡洋; 邱迪; 匡春香; 张艳; 王剑波


    A facile synthesis of various allyl sulfonamides based on imidation of allyl sulfides with chloramine-T and sub- sequent [2,3]-sigmatropic rearrangement has been achieved without metal catalysts. The reaction completes smoothly within 10 min, providing excellent yields in environment friendly solvent of alcohol. Functional groups such as bromine, hydroxyl, protected amido and aldehyde are tolerant under this condition.

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


    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.

  7. Catalytic Effect of Cetyltrimethylammonium Bromide on the Oxidation of Triethylene glycol by Chloramine-T in Acidic Medium

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    Vandana Sharma


    Full Text Available The kinetics and mechanism of cetyltrimethylammonium bromide catalyzed oxidation of triethylene glycol [2,2'-ethylene diqxybis(ethanol] by chloramine-T in acidic acid medium have been investigated. The reaction is first order dependence on chloramine-T and fractional order for triethylene glycol with excess concentration of other reactants. The catalytic effect due to cetyletrimethylammonium bromide has been studied. The small salt effect and increase in the reaction rate with increasing dielectric constant suggest the involvement of neutral molecule in the rate-determining step. The addition of p-toluene sulfonamide retards the reaction rate. The effect of chloride ion on the reaction also studied. The effect of temperature on the reaction has been investigated in the temperature range 313-333K and thermodynamic parameters were calculated from the Arrhenious plot. A tentative mechanism consistent with the experimental results has been proposed.

  8. 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


    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.

  9. Depletion of the chloramine-T marker residue, para-toluenesulfonamide, from skin-on fillet tissue of hybrid striped bass, rainbow trout, and yellow perch (United States)

    Meinertz, J.R.; Stehly, G.R.; Greseth, Shari L.; Gaikowski, M.P.; Gingerich, W.H.


    Waterborne exposure to n-sodium-n-chloro-p-toluenesulfonamide (chloramine-T) is an effective treatment for controlling fish mortalities caused by bacterial gill disease (BGD). Currently, data are being generated to gain United States Food and Drug Administration (FDA) approval for the use of chloramine-T in aquaculture. As part of the data required for an approval, depletion of the chloramine-T marker residue (para-toluenesulfonamide [p-TSA]) from the edible fillet tissue of exposed fish must be determined. Hybrid striped bass (Morone saxatilis??Morone chrysops; mean weight 357 g), rainbow trout (Oncorhynchus mykiss; mean weight 457 g), and yellow perch (Perca flavescens; mean weight 144 g) were exposed to 20 mg/l of chloramine-T for 60 min on 4 consecutive days (the most aggressive treatment expected for approved use in the United States). Groups of fish (n=15 or 19) were sampled immediately after the last treatment and periodically through 48 or 168 h after the treatment phase. Duplicate subsamples of skin-on fillet tissue from each fish were analyzed for p-TSA. Mean p-TSA concentrations in fillet tissue from fish sampled immediately after the last treatment were 142 ng/g (hybrid striped bass), 97 ng/g (rainbow trout), and 150 ng/g (yellow perch). Mean p-TSA concentrations at terminal sample times were 94 (168 h; hybrid striped bass), 74 (48 h; rainbow trout), and 35 ng/g (168 h; yellow perch). The half-lives of p-TSA in fillet tissue from fish near or at market size were 11.4 (hybrid striped bass), 4.3 (rainbow trout), and 3.2 days (yellow perch).

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

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


    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.

  11. Effects of two water disinfectants (chloramine T and peracetic acid) on the epidermis and gills of Garra rufa used in human ichthyotherapy. (United States)

    Sirri, R; Zaccaroni, A; Di Biase, A; Mordenti, O; Stancampiano, L; Sarli, G; Mandrioli, L


    Doctor fish (Garra rufa) have recently been used for aesthetic purposes and as a medical treatment in patients with psoriasis (ichthyotherapy). For this particular kind of human therapy it is essential to guarantee adequate hygienic conditions for both people and fish. The aim of this study was to test two concentrations of water disinfectants, chloramine T and peracetic acid, on Garra rufa to ascertain possible exposure damage to the epidermis and gills. Fish were exposed to 2 mg/l and 10 mg/l of chloramine T and to 15 microl/l and 45 microl/l of peracetic acid in a 40-minute static bath up to six times a day for one week. The epidermis and gills were checked for histological changes and the number of epidermal mucous cells, club cells and taste buds were quantified; mucous cells were also characterized histochemically to detect alterations in mucin production. No mortality or severe histological changes were found in treated or control fish. Cell count showed a significant increase (p peracetic acid independently of the dose. Club cell number showed a significant (p peracetic acid (mean 78.17 +/- 10.5) compared to controls (mean 107.0 +/- 19.2). Histochemical evaluation of mucous cells did not reveal changes in mucin type in fish exposed to the two disinfectants. The results suggest a good tolerability of Garra rufa to the two disinfectants at the concentrations tested.

  12. 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

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    Kanakapura Basavaiah


    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

  13. Kinetics and Mechanism of Oxidation of Diethyl Ether by Chloramine-T in Acidic Medium

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    Y. I. Hassan


    Full Text Available The kinetics of oxidation of diethyl ether (DE with sodium N-chloro-p-toluenesulphonamide (CAT in hydrochloric acid solution has been studied at (313°K.The reaction rate show a first order dependence on [CAT] and fractional order dependence on each [DE] and [H+] .The variation of ionic strength of the medium has no significant effect on the reaction rate , addition of p-toluenesulphonamide (p-TSA affects the reaction rate marginally the rate increased with decreasing dielectric constant of the medium , the stochiometry of the reaction was found to be 1:2 and oxidation products were identified , A Michaelis – Menten type mechanism has been suggested to explain the results.The equilibrium and the decomposition constants of CAT – diethyl ether complex have been evaluated. Thermodynamic parameters were computed by studying reaction at temperatures range ( 308 – 323°K for the rate limiting step and for the observed first order constants by the linear Arrhenius plot. The mechanism proposed and the derived rate law are consistent with observed kinetics.

  14. Study of Effect of Variation of Ionic Strength of the Medium on Velocity Constant of Ru(Iii Catalyzed Oxidation of Hydroxy Benzoic Acids by Chloramine-T in Acidic Medium

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    Dr. Parmod Kumar


    Full Text Available In the present study the effect of variation of ionic strength of the medium on rate of oxidation of hydroxy benzoic acids as its effect provides some clues regarding the nature of reactive species involved in the rate determining step. In order to realize the above aim the experiments has been carried out which ultimately concluded that ionic strength variation does not change the reaction velocity constant significantly.

  15. 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


    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.

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

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    D’Alò Simona


    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.

  17. Is nothing sacred? (United States)

    Hoffman, G.L.


    N-sodium-N-chloro-rho-toluenesulfonamide (chloramine-T) effectively controls bacterial gill disease (BGD) in cultured fishes, BGD, a common disease of hatchery-reared salmonids, causes more fish losses than any other disease among these species, This study describes a liquid chromatographic (LC) method that is capable of direct, simultaneous analysis of chloramine-T and its primary degradation product, rho-toluenesulfonamide (rho-TSA), in water. The procedure involves reversed-phase (C-18) LC analysis with ion suppression, using 0.01 M phosphate buffer at pH 3. The mobile phase is phosphate buffer-acetonitrile (60 + 40) at 1 mL/min. Both chemicals can be detected with a UV spectrophotometer at 229 nm; the method is linear up to 40 mg, chloramine-T or rho-TSA/L. Mean recoveries were 96.4 +/- 6.1% for water samples fortified with 0.03 mg chloramine-T/L and 95.3 +/- 4.6% for water samples fortified with 0.005 mg rho-TSA/L. Limits of detection without sample enrichment for chloramine-T and rho-TSA are 0.01 mg/L and 0.001 mg/L, respectively.

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

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    K S Ashwitha Rai


    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Press, O.W.


    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.

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

    Indian Academy of Sciences (India)



    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.

  1. Changes in the Expression of Biofilm-Associated Surface Proteins in Staphylococcus aureus Food-Environmental Isolates Subjected to Sublethal Concentrations of Disinfectants. (United States)

    Cincarova, Lenka; Polansky, Ondrej; Babak, Vladimir; Kulich, Pavel; Kralik, Petr


    Sublethal concentrations (sub-MICs) of certain disinfectants are no longer effective in removing biofilms from abiotic surfaces and can even promote the formation of biofilms. Bacterial cells can probably adapt to these low concentrations of disinfectants and defend themselves by way of biofilm formation. In this paper, we report on three Staphylococcus aureus biofilm formers (strong B+++, moderate B++, and weak B+) that were cultivated with sub-MICs of commonly used disinfectants, ethanol or chloramine T, and quantified using Syto9 green fluorogenic nucleic acid stain. We demonstrate that 1.25-2.5% ethanol and 2500 μg/mL chloramine T significantly enhanced S. aureus biofilm formation. To visualize differences in biofilm compactness between S. aureus biofilms in control medium, 1.25% ethanol, or 2500 μg/mL chloramine T, scanning electron microscopy was used. To describe changes in abundance of surface-exposed proteins in ethanol- or chloramine T-treated biofilms, surface proteins were prepared using a novel trypsin shaving approach and quantified after dimethyl labeling by LC-LTQ/Orbitrap MS. Our data show that some proteins with adhesive functions and others with cell maintenance functions and virulence factor EsxA were significantly upregulated by both treatments. In contrast, immunoglobulin-binding protein A was significantly downregulated for both disinfectants. Significant differences were observed in the effect of the two disinfectants on the expression of surface proteins including some adhesins, foldase protein PrsA, and two virulence factors.

  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


    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. Hydrogen peroxide as pre-treatment stressor in experimental immer-sion challenge of rainbow trout fry with Flavobacterium psychrophilum

    DEFF Research Database (Denmark)

    Henriksen, Maya Maria Mihályi; Madsen, Lone; Dalsgaard, Inger


    . Non-medical therapeutic substances are routinely used against pathogens in aquacultures, including copper sulphate, chloramine-T, sodium carbonates, sodium chloride, formalin and hydrogen peroxide (H2O2). One of the more successful immersion models used formalin as a stressor, but a less harmful...

  4. The dinoflagellates Pfiesteria shumwayae and Luciella masanensis cause fish kills in recirculation fish farms in Denmark

    DEFF Research Database (Denmark)

    Moestrup, Øjvind; Hansen, Gert; Daugbjerg, Niels;


    are apparently the first RAS farms in which serious fish kills have been reported. In the marine farm (Luciella) fish mortality increased dramatically despite treatment of the water with peracetic acid and chloramine-T. The plant was temporarily closed down pending investigation into the cause of mortality...

  5. Linking water treatment practices and fish welfare

    DEFF Research Database (Denmark)

    Zubiaurre, Claire; Pedersen, Lars-Flemming


    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...

  6. Treatment of trichodiniasis in eel ( Anguilla anguilla ) reared in recirculation systems in Denmark : alternatives to formaldehyde

    DEFF Research Database (Denmark)

    Madsen, H.C.K.; Buchmann, Kurt; Mellergaard, Stig


    parasiticidal effect: acriflavin (25 ppm), bithionol (0.1 ppm), chloramine T (50 ppm), Detarox AP(R) (45 ppm), malachite green (1 ppm), raw garlic (200 ppm), potassium permanganate (20 ppm) and Virkon PF(R) vet. (20 ppm). Preliminary screening revealed that the anthelmintic, bithionol, and the decomposable...

  7. 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)


    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.

  8. 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)


    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.

  9. New aquaculture drugs under FDA review (United States)

    Bowker, James D.; Gaikowski, Mark P.


    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.

  10. Changes in the Expression of Biofilm-Associated Surface Proteins in Staphylococcus aureus Food-Environmental Isolates Subjected to Sublethal Concentrations of Disinfectants

    Directory of Open Access Journals (Sweden)

    Lenka Cincarova


    Full Text Available Sublethal concentrations (sub-MICs of certain disinfectants are no longer effective in removing biofilms from abiotic surfaces and can even promote the formation of biofilms. Bacterial cells can probably adapt to these low concentrations of disinfectants and defend themselves by way of biofilm formation. In this paper, we report on three Staphylococcus aureus biofilm formers (strong B+++, moderate B++, and weak B+ that were cultivated with sub-MICs of commonly used disinfectants, ethanol or chloramine T, and quantified using Syto9 green fluorogenic nucleic acid stain. We demonstrate that 1.25–2.5% ethanol and 2500 μg/mL chloramine T significantly enhanced S. aureus biofilm formation. To visualize differences in biofilm compactness between S. aureus biofilms in control medium, 1.25% ethanol, or 2500 μg/mL chloramine T, scanning electron microscopy was used. To describe changes in abundance of surface-exposed proteins in ethanol- or chloramine T-treated biofilms, surface proteins were prepared using a novel trypsin shaving approach and quantified after dimethyl labeling by LC-LTQ/Orbitrap MS. Our data show that some proteins with adhesive functions and others with cell maintenance functions and virulence factor EsxA were significantly upregulated by both treatments. In contrast, immunoglobulin-binding protein A was significantly downregulated for both disinfectants. Significant differences were observed in the effect of the two disinfectants on the expression of surface proteins including some adhesins, foldase protein PrsA, and two virulence factors.

  11. Synthesis of iodobuprenorphine for use in radioimmunoassay. [Analgesic

    Energy Technology Data Exchange (ETDEWEB)

    Debrabandere, L.; Boven, M. van; Daenens, P. (Leuven Univ. (Belgium). Lab. of Toxicology)


    The synthesis of [[sup 125]I]iodobuprenorphine is described. This compound has been used as a tracer molecule in the development of a new radioimmunoassay for buprenorphine. The parameters that effect the direct iodination of buprenorphine using sodium [[sup 127]I]iodide, in the presence of either chloramine-T or Iodo-Beads, were studied. The structure of iodobuprenorphine was confirmed by mass spectrometry. (author).

  12. Peracetic acid products expand sanitizing, organic water treatment options


    Pedersen, Lars-Flemming; Jokumsen, Alfred; Larsen, Villy J.; Henriksen, Niels Henrik


    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 optimizing treatment protocols a challenge. Continuous low-dose applications seem to be a promising solution. PAA is among the few disinfectants approved for organic aquaculture

  13. Assays for calcitonin receptors

    Energy Technology Data Exchange (ETDEWEB)

    Teitelbaum, A.P.; Nissenson, R.A.; Arnaud, C.D.


    The assays for calcitonin receptors described focus on their use in the study of the well-established target organs for calcitonin, bone and kidney. The radioligand used in virtually all calcitonin binding studies is /sup 125/I-labelled salmon calcitonin. The lack of methionine residues in this peptide permits the use of chloramine-T for the iodination reaction. Binding assays are described for intact bone, skeletal plasma membranes, renal plasma membranes, and primary kidney cell cultures of rats. Studies on calcitonin metabolism in laboratory animals and regulation of calcitonin receptors are reviewed.

  14. 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)


    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).

  15. Synthesis of new series of 5,6-dihydro-4H-1,2-oxazines via hetero Diels-Alder reaction and evaluation of antimicrobial activity. (United States)

    Manjula, M K; Rai, K M L; Gaonkar, S L; Raveesha, K A; Satish, S


    A new series of 5,6-dihydro-4H-1,2-oxazines were synthesized via hetero Diels-Alder reaction of alpha-nitrosoolefins with alkenes. alpha-Nitrosoolefins were generated from ketoximes by the action of chloramine-T. The newly synthesized compounds were characterized with IR, NMR, elemental analysis and screened for their antimicrobial activity; they exhibited excellent antimicrobial activity. The minimal inhibitory concentration of the compounds was in the range of 10-35 microg ml(-1) for bacteria and 10-40 microg ml(-1) for fungi.

  16. Synthesis and in vitro assessment of antifungal activity of oximes, oxime ethers and isoxazoles.


    Diaz-Velandia, John; Laboratorio de síntesis orgánica, Departamento de Química. Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia.; Durán-Díaz, Natalia; Laboratorio de síntesis orgánica, Departamento de Química. Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia.; Robles-Camargo, Jorge; Laboratorio de síntesis orgánica, Departamento de Química. Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia.; Loaiza, Alix Elena; Laboratorio de síntesis orgánica, Departamento de Química. Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia


    Objective. To synthesize and carry out a preliminary evaluation of the in vitro antifungal activity of oximes, oxime ethers and isoxazoles. Materials and methods. Oximes were synthesized from aldehydes or ketones with NH2OH.HCl and K2CO3. Oxime ethers were prepared by alkylation of oximes with propargyl bromide or 2-bromobenzyl bromide, using NaOH as base and acetone as solvent. The isoxazoles were obtained by 1,3-dipolar cycloadditions using ceric ammonium nitrate (CAN), chloramine T (CA...

  17. 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


    .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......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...

  18. Methods to eradicate soft tunic syndrome (STS)-causing protozoa Azumiobodo hoyamushi, the highly infectious parasite from the edible ascidian (Halocynthia roretzi)


    Ji-Hoon Lee; Jae-Geun Lee; Seung-Ryul Zeon; Kyung-Il Park; Kwan Ha Park


    Abstract Although soft tunic syndrome (STS) in the ascidian is a serious disease, helpful measures have yet not been established. It was examined in this study by applying aniti-parasitic drugs to eradicate the causative protozoa Azumiobodo hoyamushi from infected ascidians. Formalin was synergistic in killing parasites in vitro when co-treated with hydrogen peroxide (H2O2) or bronopol, but not with chloramine-T or povidone-iodine (PVP-I), when tested with in vitro parasite culture. The syner...

  19. 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))


    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.

  20. 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:; 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)


    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.

  1. Two in-vivo protocols for testing virucidal efficacy of handwashing and hand disinfection. (United States)

    Steinmann, J; Nehrkorn, R; Meyer, A; Becker, K


    Whole-hands and fingerpads of seven volunteers were contaminated with poliovirus type 1 Sabin strain in order to evaluate virucidal efficacy of different forms of handwashing and handrub with alcohols and alcohol-based disinfectants. In the whole-hand protocol, handwashing with unmedicated soap for 5 min and handrubbing with 80% ethanol yielded a log reduction factor (RF) of > 2, whereas the log RF by 96.8% ethanol exceeded 3.2. With the fingerpad model ethanol produced a greater log RF than iso- or n-propanol. Comparing five commercial hand disinfectants and a chlorine solution (1.0% chloramine T-solution) for handrub, Desderman and Promanum, both composed of ethanol, yielded log RFs of 2.47 and 2.26 respectively after an application time of 60 s, similar to 1.0% chloramine T-solution (log RF of 2.28). Autosept, Mucasept, and Sterillium, based on n-propanol and/or isopropanol, were found to be significantly less effective (log RFs of 1.16, 1.06 and 1.52 respectively). A comparison of a modified whole-hand and the fingerpad protocol with Promanum showed similar results with the two systems suggesting both models are suitable for testing the in-vivo efficacy of handwashing agents and hand disinfectants which are used without any water.

  2. 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.


    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.

  3. Optimisation of an Advanced Oxidation Protein Products Assay: Its Application to Studies of Oxidative Stress in Diabetes Mellitus

    Directory of Open Access Journals (Sweden)

    Emma L. Taylor


    Full Text Available Advanced oxidation protein products (AOPP are reportedly elevated in the plasma of patients with a number of diseases, including diabetes mellitus, that involve oxidative stress. However, the accurate measurement of AOPP in human plasma is hampered by the formation of a precipitate following the addition of potassium iodide and glacial acetic acid according to the published assay procedure. Here we describe a modification of the AOPP assay which eliminates interference by precipitation and provides a robust, reliable, and reproducible protocol for the measurement of iodide oxidising capacity in plasma samples (intra-assay CV 1.7–5.3%, interassay CV 5.3–10.5%. The improved method revealed a significant association of AOPP levels with age (p<0.05 and hypertension (p=0.01 in EDTA-anticoagulated plasma samples from 52 patients with diabetes and 38 nondiabetic control subjects, suggesting a possible link between plasma oxidising capacity and endothelial and/or vascular dysfunction. There was no significant difference between AOPP concentrations in diabetic (74.8 ± 7.2 μM chloramine T equivalents and nondiabetic (75.5 ± 7.0 μM chloramine T equivalents individuals.

  4. 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))


    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.

  5. Radioimmunoassay of secretin in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Bonora, G.; Vezzadini, P.; Toni, R.; Labo, G. (Bologna Univ. (Italy))


    A sensitive radioimmunoassay for secretin has been developed. Antisera were raised against synthetic porcine secretin coupled to bovine serum albumin. N-..cap alpha..-desaminotyrosyl-..beta..-alanyl secretin was radioiodinated by a slight modification of the chloramine-T method. Pure synthetic porcine secretin was used as a standard. Free and bound hormone were separated by dextran-coated charcoal. No cross-reactivity was found with structurally and physiologically related peptides. The sensitivity of the assay was high enough to measure fasting secretin levels in human serum. Patients with acute or chronic pancreatitis had mean serum secretin concentration not significantly different from healthy subjects. In patients with pancreatic carcinoma the mean serum secretin concentration was significantly lower than in healthy subjects, although a wide overlap of the two groups was evident.

  6. Linking water treatment practices and fish welfare

    DEFF Research Database (Denmark)

    Zubiaurre, Claire; Pedersen, Lars-Flemming


    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...... 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....... 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...


    Institute of Scientific and Technical Information of China (English)

    刘宗柱; 王金宝; 徐永立; 王勇; 张培军


    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.


    Institute of Scientific and Technical Information of China (English)


    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.

  9. Synthesis and preliminary biological evaluation of [{sup 123}I]Me{sub 2}Pyr, a new potential ligand for imaging of central cannabinoid CB{sub 1} receptors

    Energy Technology Data Exchange (ETDEWEB)

    Gielow, P. [Department of Nuclear Medicine, University School of Medicine, Carl-Neuberg-Str. 1, D-30625 Hannover (Germany)]. E-mail:; Klinge, P. [International Neuroscience Institute, Alexis Carrel Str. 4, D-30625 Hannover (Germany); Knapp, W.H. [Department of Nuclear Medicine, University School of Medicine, Carl-Neuberg-Str. 1, D-30625 Hannover (Germany); Berding, G. [Department of Nuclear Medicine, University School of Medicine, Carl-Neuberg-Str. 1, D-30625 Hannover (Germany)


    A synthesis of 1-(2,4-dichlorophenyl)-5-(4-[{sup 123}I]iodophenyl)-4-methyl-1H-pyrazole -3-carboxylic acid N',N'-dimethyl-hydrazide ([{sup 123}I]Me{sub 2}Pyr), a new radioiodinated analogue of the high-affinity cannabinoid CB{sub 1} receptor antagonist SR141716A, is described. Labelling was achieved by radioiododestannylation of the tributylstannyl precursor with [{sup 123}I]iodide in the presence of chloramine T. HPLC purification afforded the labelled product in 48% radiochemical yield. Preliminary rat brain biodistribution studies with the {sup 125}I labelled compound revealed high uptake in the substantia nigra, the globus pallidus externus and the cerebellum, which is consistent with the known distribution of CB{sub 1} receptors.

  10. Microwave-Assisted Solution Phase Synthesis of Novel 2-{4-[2-(N-Methyl-2-pyridylaminoethoxy]phenyl}-5-Substituted 1,3,4-Oxadiazole  Library

    Directory of Open Access Journals (Sweden)

    Santosh L. Gaonkar


    Full Text Available A new series of 2-{4-[2-(N-methyl-2-pyridylaminoethoxy]phenyl}-5-substituted 1,3,4-oxadiazoles were synthesized by the oxidative cyclization of hydrazones derived from 4-[2-(methylpyridin-2-ylaminoethoxy]benzaldehyde and aryl hydrazines using chloramine-T as an efficient catalyst. All steps were assisted by microwave irradiation. Microwave enables all these reactions to be simple, rapid, high yielding, and avoid chromatograph purification and led environmentally benign total synthesis of focused oxadiazole library. All the synthesized compounds were isolated in good yield and characterized by 1H NMR, 13C NMR, and elemental analyses. The title compounds represent a novel class of biologically active heterocycles.

  11. Synthesis and antimicrobial studies of a new series of 2-[4-[2-(5-ethylpyridin-2-yl)ethoxy]phenyl]-5-substituted-1,3,4-oxadiazoles. (United States)

    Gaonkar, S L; Rai, K M L; Prabhuswamy, B


    A series of novel 2-[4-[2-(5-ethylpyridin-2-yl)ethoxy]phenyl]-5-substituted-1,3,4-oxadiazoles were synthesized by the oxidative cyclisation of hydrazones derived from 4-[2-(5-ethylpyridin-2-yl)ethoxy]benzaldehyde and aroylhydrazines using chloramine-T as oxidant. IR, NMR and elemental analysis characterized the newly synthesized compounds. The synthesized compounds were evaluated for their antimicrobial activity and were compared with standard drugs. The compounds demonstrated potent to weak antimicrobial activity. Out of the compounds studied, compounds 8c and 8d showed significant inhibition. Compounds 8b, 8f, 8k and 8e showed moderate activity. The minimum inhibitory concentration of the compounds was in the range of 8-26 microg ml(-1) against bacteria and 8-24 microg ml(-1) against fungi. The title compounds represent a novel class of potent antimicrobial agents.

  12. Mild radioiodination of insulin with dichloramine-T

    Energy Technology Data Exchange (ETDEWEB)

    Jyotsna, T.; Pillai, M.R.A.; Gupte, J.H.; Mani, R.S.


    Iodination of insulin using N-dichloro-p-toluene sulphonamide(dichloramine-T) has been standardised. Dichloramine-T, a water insoluble derivative of chloramine-T showed excellent properties as an iodination reagent, for the preparation of radiolabeled insulin for use in radioimmunoassay. Iodination using dichloramine-T could be done at as low as 0.5-2 of the reagent, and at this concentration the molar ratio worked out to 1:3 to 1:12 (protein: dichloramine-T). Iodination yields of greater than 90% were obtained at pH 5-7. Evaluation of the iodinated tracer for suitability in radioimmunoassay was carried out by estimating radiochemical purity, immunological purity, non specific binding and stability on storage.

  13. Protein aggregation and degradation during iodine labeling and its consequences for protein adsorption to biomaterials

    DEFF Research Database (Denmark)

    Holmberg, Maria; Jensen, Karin Bagger Stibius; Ndoni, Sokol;


    were analyzed through chromatography and spectroscopy techniques. Results show that the iodine labeling method using chloramine-T (CAT) as an oxidizing agent can cause both enhanced aggregation and fragmentation of proteins. Albumin shows an enhanced tendency to aggregate after iodine labeling using......Protein adsorption on modified and unmodified polymer surfaces investigated through radiolabeling experiments showed a tendency for higher than expected albumin and immunoglobulin G (IgG) adsorption. Possible enhanced protein aggregation and degradation caused by the iodine labeling method used...... of protein adsorption should be used with caution and preferably should be validated by a label-free methodology for each combination of radio-label and protein. The results obtained in this study can be used to optimize investigation of protein adsorption on surfaces of materials for biomedical devices. (c...

  14. Sensitivity of juvenile striped bass to chemicals used in aquaculture

    Energy Technology Data Exchange (ETDEWEB)

    Bills, T.D.; Marking, L.L.; Howe, G.E.


    Efforts to restore anadromous striped bass (Morone saxatilis) populations by the U.S. Fish and Wildlife Service and other agencies over the past 20 years have concentrated on hatchery culture to supplement dwindling natural reproduction. Adult fish captured for artificial spawning are stressed by handling and crowding in rearing ponds and are often exposed to therapeutants, anesthetics, disinfectants, and herbicides used in fish culture. The authors determined the toxicity of 17 fishery chemicals (chloramine-T, erythromycin, formalin, Hyamine 3500, Roccal, malachite green, sulfamerazine, benzocaine, etomidate, Finquel (MS-222), metomidate, quinaldine sulfate, chlorine, potassium permanganate, Aquazine, copper sulfate, and Rodeo) to striped bass fry (average weight = 1 g) in reconstituted water (total hardness 40 mg/L) at 12 degrees C. The 96-h LC50's (concentration calculated to produce 50% mortality in a population) ranged from 0.129 mg/L for malachite green to 340 mg/L for erythromycin.

  15. Chemical modification of amino acid residues in glycerinated Vorticella stalk and Ca(2+)-induced contractility. (United States)

    Kono, R; Ochiai, T; Asai, H


    The glycerinated stalk of the peritrich ciliate Vorticella, was treated with various reagents to chemically modify the amino acid residues. The influences of these modifcations on spasmoneme contractility were investigated. First, it was confirmed that the spasmoneme contraction is not inhibited by alteration of SH groups. It was also demonstrated that chemical modification of methionine and tryptophan residues abolishes spasmoneme contractility. The reagents used for chemical modification were N-bromosuccinimide (NBS), chloramine T, and 2-hydroxy-5-nitrobenzyl bromide (HNBB), which abolished spasmoneme contractility at concentrations of 40-50 microM, 200-300 microM, and 4 mM, respectively. These results suggest that, along with Ca2+ binding proteins, there are other as yet to be identified proteins involved in contractility.

  16. 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)


    Purified human liver ..cap alpha..-L-fucosidase (EC has been radioiodinated by a chloramine-T procedure to a specific activity of 3.7 x 10/sup 6/ dpm/ 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.

  17. Preparation of 125I-protein A usable for up to 10 months in immunoassays

    DEFF Research Database (Denmark)

    Dyrberg, T; Billestrup, Nils


    Chloramine-T iodination of protein A from Staphylococcus aureus and gel electrophoretic purification of the iodination mixture results in a stable tracer of high specific and functional activity. Following repeated gel electrophoresis of the tracer only a single component was observed. The specific...... activity of the 125I-protein A was between 30 and 55 muCi/micrograms. The binding of 125I-protein A to rabbit immunoglobulin exceeded 90% and the tracer competed effectively with unlabelled protein A in binding to cells incubated with sera containing surface antibodies. Storage of the tracer for up to 46...... weeks resulted in a moderate decrease in maximal binding to immunoglobulin (from 91% to 64%), in TCA precipitable radioactivity (from 97% to 80%) and an approx. 30% decrease in the ability to detect cell bound immunoglobulin. It is concluded that gel electrophoretic purification of 125I-protein...

  18. Reactive species modify NaV1.8 channels and affect action potentials in murine dorsal root ganglion neurons. (United States)

    Schink, Martin; Leipold, Enrico; Schirmeyer, Jana; Schönherr, Roland; Hoshi, Toshinori; Heinemann, Stefan H


    Dorsal root ganglion (DRG) neurons are important relay stations between the periphery and the central nervous system and are essential for somatosensory signaling. Reactive species are produced in a variety of physiological and pathophysiological conditions and are known to alter electric signaling. Here we studied the influence of reactive species on the electrical properties of DRG neurons from mice with the whole-cell patch-clamp method. Even mild stress induced by either low concentrations of chloramine-T (10 μM) or low-intensity blue light irradiation profoundly diminished action potential frequency but prolonged single action potentials in wild-type neurons. The impact on evoked action potentials was much smaller in neurons deficient of the tetrodotoxin (TTX)-resistant voltage-gated sodium channel NaV1.8 (NaV1.8(-/-)), the channel most important for the action potential upstroke in DRG neurons. Low concentrations of chloramine-T caused a significant reduction of NaV1.8 peak current and, at higher concentrations, progressively slowed down inactivation. Blue light had a smaller effect on amplitude but slowed down NaV1.8 channel inactivation. The observed effects were less apparent for TTX-sensitive NaV channels. NaV1.8 is an important reactive-species-sensitive component in the electrical signaling of DRG neurons, potentially giving rise to loss-of-function and gain-of-function phenomena depending on the type of reactive species and their effective concentration and time of exposure.

  19. Labeling of the Recombinant Streptokinase Using Iodine-131 as a New Thrombolytic Agent

    Directory of Open Access Journals (Sweden)

    I. Daruwati


    Full Text Available Stroke and acute myocardial infarction is a disease with the highest mortality in the world. WHO has estimated in 2008, 30% of deaths from heart disease and more than 80% of this cases have been occurred in developing countries. Streptokinase (SK as an effective thrombolytic agent has been used as a drug of choice for about forty years ago. SK is plasminogen (PG activator that converts plasminogen to active protease, called plasmin (PN which degrades fibrin to soluble products. Recombinant streptokinase (SKA from genetic engineering has been developed at School of Pharmacy ITB to reduce or eliminate immunogenicity of SK. However, scientific disclosures relating to dynamic and its kinetic studies in the body have still to be proven. One method that can explain this phenomenon is the pharmacological studies using radionuclide labeled compounds. Radioiodine labeled compound is used extensively and most suitable for biological studies. This paper describes the preparation of 131I-SKA and its characterization. The labeling conditions of SKA, such as chloramine-T as an oxidizing agent, amount of SKA, incubation time, and size of resin to purify the labeling yield have been observed. The result showed that the optimum condition of labeling (35.11% was obtained using 10 µg of chloramine-T and 60 seconds of incubation time. The highest radiochemical purity (97.46 ± 1.14% has also been obtained by passing through the resin chromatography column using 100 mg Dowex 1x8, size 50-100 mesh. The characterization of 131I-SKA with SDS PAGE method and autoradiography showed the similar performance with unlabeled-SKA

  20. An experimental study on (131I-CHIBA-1001: a radioligand for α7 nicotinic acetylcholine receptors.

    Directory of Open Access Journals (Sweden)

    Lei Yin

    Full Text Available OBJECTIVE: The α7 nicotinic acetylcholine receptors (nAChRs play a vital role in the pathophysiology of neuropsychiatric diseases such as Alzheimer's disease and depression. However, there is currently no suitable positron emission tomography (PET or Single-Photon Emission Computed Tomography (SPECT radioligands for imaging α7 nAChRs in brain. Here our aim is to radiosynthesize a novel SPECT radioligand (131I-CHIBA-1001 for whole body biodistribution study and in vivo imaging of α7 nAChRs in brain. METHOD: (131I-CHIBA-1001 was radiosynthesized by chloramine-T method. Different conditions of reaction time and temperature were tested to get a better radiolabeling yield. Radiolabeling yield and radiochemical purities of (131I-CHIBA-1001 were analyzed by thin layer chromatography (TLC and high-performance liquid chromatography (HPLC system. Whole body biodistribution study was performed at different time points post injection of (131I-CHIBA-1001 in KM mice. Monkey subject was used for in vivo SPECT imaging in brain. RESULT: The radiolabeling yield of (131I-CHIBA-1001 reached 96% within 1.5∼2.0 h at 90∼95°C. The radiochemical purity reached more than 99% after HPLC purification. (131I-CHIBA-1001 was highly stable in saline and fresh human serum in room temperature and 37°C separately. The biodistribution data of brain at 15, 30, and 60 min were 11.05±1.04%ID/g, 8.8±0.04%ID/g and 6.28±1.13%ID/g, respectively. In experimental SPECT imaging, the distribution of radioactivity in the brain regions was paralleled with the distribution of α7 nAChRs in the monkey brain. Moreover, in the blocking SPECT imaging study, the selective α7 nAChR agonist SSR180711 blocked the radioactive uptake in the brain successfully. CONCLUSION: The CHIBA-1001 can be successfully radiolabeled with (131I using the chloramine-T method. (131I-CHIBA-1001 can successfully accumulate in the monkey brain and image the α7 acetylcholine receptors. (131I-CHIBA-1001 can be a

  1. Synthesis of [{sup 123}I]iodine labelled imidazo[1,2-b] pyridazines as potential probes for the study of peripheral benzodiazepine receptors using SPECT

    Energy Technology Data Exchange (ETDEWEB)

    Katsifis, A.; Mattner, F.; Dikic, B. [Radiopharmaceuticals Div. ANSTO, Menai, NSW (Australia); Barlin, G. [Div. of Neurosciences, John Curtin School of Medical Research, Australian National Univ., Canberra (Australia)


    The pyridazines 3-acetamidomethyl-6-chloro-2-(4'-iodophenyl)imidazo[1,2-b]pyridazine 1 (IC{sub 50} = 1.6 nM) and 3-benzamidomethyl-6-iodo-2-(4'-t-butylphenyl)imidazo[1,2-b] pyridazine 2 (IC{sub 50} = 4.2 nM), are high affinity and selective ligands for the peripheral benzodiazepine receptors (PBR) compared to the central benzodiazepine counterparts. The [{sup 123}I]1 and [{sup 123}I]2 labelled analogues of these compounds were subsequently synthesised for the potential study of the PBR in vivo using SPECT. Radioiodination of [{sup 123}I]1 was achieved by iododestannylation of the corresponding tributyl tin precursor with Na[{sup 123}I] in the presence of peracetic acid or chloramine-T and the product isolated by C-18 RP HPLC. Radioiodination of [{sup 123}I]2 was achieved by copper assisted bromine [{sup 123}I]iodine exchange of the corresponding bromo precursor in the presence of acetic acid and sodium bisulfate as reducing agent at 200 C. Purification of the crude products were achieved by semi-preparative C-18 RP HPLC to give the products in radiochemical yields > 90%. The products were obtained in > 97% chemical and radiochemical purity and with specific activities > 180 GBq/{mu}mol. (orig.)

  2. Bromine-76 and carbon-11 labelled NNC 13-8199, metabolically stable benzodiazepine receptor agonists as radioligands for positron emission tomography (PET)

    Energy Technology Data Exchange (ETDEWEB)

    Foged, C. [Karolinska Institutet, Department of Clinical Neuroscience, Psychiatry Section, Karolinska Hospital, Stockholm (Sweden)]|[Novo Nordisk A/S, Health Care Discovery and Development, Maaloev (Denmark); Halldin, C.; Pauli, S.; Suhara, T.; Swahn, C.G.; Karlsson, P.; Farde, L. [Karolinska Institutet, Department of Clinical Neuroscience, Psychiatry Section, Karolinska Hospital, Stockholm (Sweden); Loc`h, C.; Maziere, B.; Maziere, M. [Service Hospitalier Frederic Joliot, CEA, Orsay (France); Hansen, H.C. [Novo Nordisk A/S, Health Care Discovery and Development, Maaloev (Denmark)


    NNC 13-8241 has recently been labelled with iodine-123 and developed as a metabolically stable benzodiazepine receptor ligand for single-photon emission computed tomography (SPECT) in monkeys and man. NNC 13-8199 is a bromo-analogue of NNC 13-8241. This partial agonist binds selectively and with subnanomolar affinity to the benzodiazepine receptors. We prepared {sup 76}Br labelled NNC 13-8199 from the trimethyltin precursor by the chloramine-T method. Carbon-11 labelled NNC 13-8199 was synthesised by N-alkylation of the nitrogen of the amide group with [{sup 11}C]methyl iodide. Positron emission tomography (PET) examination with the two radioligands in monkeys demonstrated a high uptake of radioactivity in the occipital, temporal and frontal cortex. In the study with [{sup 76}Br]NNC 13-8199, the monkey brain uptake continued to increase until the time of displacement with flumazenil at 215 min after injection. For both radioligands the radioactivity in the cortical brain regions was markedly reduced after displacement with flumazenil. More than 98% of the radioactivity in monkey plasma represented unchanged radioligand 40 min after injection. The low degree of metabolism indicates that NNC 13-8199 is metabolically much more stable than hitherto developed PET radioligands for imaging of benzodiazepine receptors in the primate brain. [{sup 76}Br]NNC 13-8199 has potential as a radioligand in human PET studies using models where a slow metabolism is an advantage. (orig.) With 8 figs., 28 refs.

  3. Development of a radioiodinated ligand for characterising. cap alpha. /sub 1/-adrenoceptors. [Pentolamine and 2 BETA-(4-hydroxyphenyl)-ethylaminomethyl)-tetralone

    Energy Technology Data Exchange (ETDEWEB)

    Adams, A.; Jarrott, B.


    Two ..cap alpha..-adrenoceptor antagonists, phentolamine and 2-(..beta..-(4-hydroxyphenyl)-ethylaminomethyl)-tetralone (BE 2254) which are phenolic derivatives were radioiodinated after chloramine-T oxidation of Na/sup 125/I and the labelled material isolated by chromatography. /sup 125/I-Phentolamine does not bind selectively to ..cap alpha..-adrenoceptors in guinea pig brain whereas the /sup 125/I-BE 2254 derivative binds rapidly, reversibly and with high affinity to these receptors with a K/sub d/ of 230 pM. At low concentrations of /sup 125/I-BE 2254 (< 100 pM) approx. 90% of the bound radioligand is specifically bound and under these conditions drug displacement studies show that the ligand binds predominantly to the ..cap alpha../sub 1/ subclass of adrenoceptors. Binding measurements to kidney and smooth muscle membrane preparations indicate that /sup 125/I-BE 2254 may also be a useful tool in the study of ..cap alpha..-adrenoceptors in peripheral tissues. The high specific activity of /sup 125/I-BE 2254 permits the use of minimal quantities of membrane material for receptor assay and ligand displacement measurements, e.g. 250 per assay tube, and this provides a significant advantage over the use of existing radioligands such as /sup 3/H-prazosin which requires approx. 40 times as much tissue.

  4. Pathological and electron microscopic studies on cold water disease among cultured rainbow trout (Oncorhynchus Mykiss Walbaum

    Directory of Open Access Journals (Sweden)

    Salah Mesalhy Aly


    Full Text Available The present study aims to investigate the etiology and pathogenesis of cold water disease among cultured rainbow trout (Oncorhynchus Mykiss Walbaum reared at low water temperature on a private farm, and propose preventive and control measures to prevent the recurrence of the disease and its spread to the neighboring farms or to other countries. The disease is caused by Flavobacterium psychrophilum and is characterized clinically by high mortality rates (25% with necrotizing skin and fin lesions. Septicemia with muscular and gill involvement is observed in severely affected cases. The histopathological and ultrastructure alterations in the infected trout explain the pathogencity of this microorganism in many organs other than skin and fins either directly by these bacteria or by its extracellular products, or indirectly via the hypoxia and ion imbalance that results from skin and gill damage. Although some antibiotics, such as trimethoprime, chloramin T and oxytetracycline, are effective on the causative agent, improvement in water quality, vaccination, increasing fish resistance and strict hygiene as well as quarantine measures would all help.

  5. Short communication: rapid preparation of preventive and therapeutic whole-killed retroviral vaccines using the microbicide taurine chloramine. (United States)

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


    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.

  6. 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


    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.

  7. Serum prostate-specific acid phosphatase: development and validation of a specific radioimmunoassay. [/sup 125/I tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Vihko, P.; Sajanti, E.; Jaenne, O.; Peltonen, L.; Vihko, R.


    We describe radioimmunoassay for human prostatic acid phosphatase (orthophosphoric-monoester phosphohydrolase (acid optimum), EC in serum, with use of monospecific antisera raised in rabbits against highly purified acid phosphatase from human prostates. The antiserum did not cross react with partly purified acid phosphatases from human spleen, erythrocytes, or synovial tissues. /sup 125/I-labeled acid phosphatase was prepared by a Chloramine T method, and the bound and free antigen was separated in the assay by use of anti-rabbit gamma-globulin raised in sheep. Uniform low nonspecific binding of the (/sup 125/I)acid phosphatase was achieved by using acid-phosphatase-free serum to prepare standard curves and diluted samples of serum with high acid phosphatase activities. Concentrations of immunoreactive acid phosphatase in the serum of healthy men ranged from <1 to 10 and for 12 patients with advanced prostatic carcinoma between 100 and 500 The concentrations of the enzyme in sera of patients with benign prostatic hyperplasia were very similar to those in sera of the reference group.

  8. 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;


    Fetal antigen (FA2) was purified from second trimester human amniotic fluid by immunospecific chromatography, gel filtration and reversed-phase chromatography. Gel filtration revealed two molecular forms of FA2 eluting at volumes corresponding to an M(r) of approximately 100 kDa and 30 kDa. SDS-P...... to that of FA2 in human skin. FA2 is a circulating form of the aminopropeptide of the alpha 1 chain of procollagen type I, and this is the first description of its isolation and structural characterization in humans. Udgivelsesdato: 1992-Dec...... 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...

  9. 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.


    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.

  10. 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.


    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.)

  11. 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


    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.

  12. {sup 99m}Tc-labeled chimeric anti-NCA 95 antigranulocyte monoclonal antibody for bone marrow imaging

    Energy Technology Data Exchange (ETDEWEB)

    Sarwar, M.; Higuchi, Tetsuya; Tomiyoshi, Katsumi [Gunma Univ., Maebashi (Japan). School of Medicine] [and others


    Chimeric mouse-human antigranulocyte monoclonal antibody (ch MAb) against non-specific cross-reacting antigen (NCA-95) was labeled with {sup 99m}Tc (using a direct method) and {sup 125}I (using the chloramine T method), and its binding to human granulocytes and LS-180 colorectal carcinoma cells expressing carcinoembryonic antigen on their surfaces, cross-reactive with anti-NCA-95 chimeric monoclonal antibody, increased in proportion to the number of cells added and reached more than 80% and 90%, respectively. In biodistribution studies, {sup 99m}Tc and {sup 125}I-labeled ch anti-NCA-95 MAb revealed high tumor uptake, and the tumor-to-blood ratio was 2.9 after 24 hours. The tumor-to-normal-organ ratio was also more than 3.0 in all organs except for the tumor-to-kidney ratio. Scintigrams of athymic nude mice confirmed the results of biodistribution studies that showed higher radioactivity in tumor and kidney of the mice administered with {sup 99m}Tc-labeled ch MAb. A normal volunteer injected with {sup 99m}Tc-labeled ch anti-NCA-95 antigranulocyte MAb showed clear bone marrow images, and a patient with aplastic anemia revealed irregular uptake in his lumbar spine, suggesting its utility for bone marrow scintigraphy and for the detection of hematological disorders, infections, and bone metastasis. (author)

  13. 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)


    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.

  14. Radiolabelling of cholera toxin

    Energy Technology Data Exchange (ETDEWEB)

    Santos, R.G.; Neves, Nicoli M.J. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil); Abdalla, L.F.; Brandao, R.L.; Etchehebehere, L. [Ouro Preto Univ., MG (Brazil). Escola de Farmacia. Lab. de Fisiologia e Bioquimica de Microorganismos; Lima, M.E. de [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Bioquimica e Imunologia; Nicoli, J.R. [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Inst. de Ciencias Biologicas. Dept. de Microbiologia


    Binding of cholera toxin to ganglioside receptors of enterocyte microvilli catalyzes the activation of adenylate cyclase causing a rise in cAMP which final result is a copious diarrhea. Saccharomyces boulardii, a nonpathogenic yeast has been used to prevent diarrhea. Although the antidiarrheic properties of S. boulardii are widely recognized, this yeast has been used on empirical basis, and the mechanism of this protective effect is unknown. The addition of cholera toxin to S. boulardii induces the raising of cAMP that triggers the activation of neutral trehalase. This suggests that toxin specifically binding to cells, is internalized and active the protein phosphorylation cascade. Our objective is labeling the cholera toxin to verify the presence of binding sites on yeast cell surfaces for the cholera toxin. Cholera toxin was radiolabelled with Na {sup 125} I by a chloramine-T method modified from Cuatrecasas and Griffiths et alii. The {sup 125} I-Cholera toxin showed a specific radioactivity at about 1000 cpm/fmol toxin. Biological activity of labeled cholera toxin measured by trehalase activation was similar to the native toxin. (author) 5 refs., 3 figs.; e-mail: nevesmj at

  15. 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)


    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.

  16. 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)


    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).

  17. 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


    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)

  18. 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


    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.

  19. 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


    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.

  20. Synthesis of substituted [{sup 123}I]imidazo[1,2-a]pyridines as potential probes for the study of the peripheral benzodiazepine receptors using SPECT

    Energy Technology Data Exchange (ETDEWEB)

    Katsifis, A.; Mattner, F.; Dikic, B.; Papazian, V. [Radiopharmaceuticals Div. R and D, ANSTO, Menai, NSW (Australia)


    The imidazo[1,2-a]pyridines N,N'-dimethyl-6-chloro-(4'-iodophenyl)imidazo[1,2-a]pyridine-3-acetamide 1. N,N'-diethyl-6-chloro-(4'-iodophenyl)imidazo[1,2-a]pyridine-3-acetamide 2, and N-methyl-6-chloro-(4'-iodophenyl)imidazo[1,2-a]pyridine-3-acetamide 3, are high affinity and selective ligands for the peripheral benzodiazepineodiazepine receptors (PBR). The [{sup 123}I]1-3 labelled analogues of these compounds were subsequently synthesised for the potential study of the PBR in vivo using SPECT. Radioiodination was achieved by iododestannylation reactions of the corresponding tributyl tin precursors with Na[{sup 123}I] in the presence of peracetic acid, chloramine-T or Iodogen. Purification of the crude product was achieved by semipreparative C-18 RP HPLC to give the products in radiochemical yields of 40-85%. The products were obtained in >97% chemical and radiochemical purity and with specific activities >80 GBq/{mu}mol. (orig.)

  1. Methods to eradicate soft tunic syndrome (STS-causing protozoa Azumiobodo hoyamushi, the highly infectious parasite from the edible ascidian (Halocynthia roretzi

    Directory of Open Access Journals (Sweden)

    Ji-Hoon Lee


    Full Text Available Abstract Although soft tunic syndrome (STS in the ascidian is a serious disease, helpful measures have yet not been established. It was examined in this study by applying aniti-parasitic drugs to eradicate the causative protozoa Azumiobodo hoyamushi from infected ascidians. Formalin was synergistic in killing parasites in vitro when co-treated with hydrogen peroxide (H2O2 or bronopol, but not with chloramine-T or povidone-iodine (PVP-I, when tested with in vitro parasite culture. The synergistic effects did not change when formalin-H2O2 (or bronopol ratios were changed. It was found that treatment periods less than 60 min achieved a sub-maximal efficacy. Increasing drug concentration while keeping 30 min period improved anti-parasitic effects. Anti-parasitic effects of formalin(F + H2O2(H were also assessed in an in vivo STS model infected with cultured parasites. It was observed that combined 50 (40F + 10H and 100 (80F +20H ppm were effective in partially preventing STS-caused mortality. In horizontally transmitted artificial STS model, significant prevention of ascidian mortality was also observed after 50 ppm. Marked reduction of living parasites were noted after drug treatments in vivo. The results provide a highly useful basis to develop a preventive or treatment measure against the currently uncontrollable STS in the ascidian.

  2. Immunoradiometric assay for quantification of prostate-specific antigen (PSA)

    Energy Technology Data Exchange (ETDEWEB)

    Ruiz, Miriam [Institute of Nuclear Sciences and Technology, Havana (Cuba). Dept.of Radiochemistry]. E-mail:


    To develop an immunoradiometric assay (IRMA) for quantification of total PSA in serum, we carried out the evaluation of a panel of 12 monoclonal antibodies (MAbs). The mAbs (CB-PSA 1 to CB-PSA 10) were developed in the Center of Genetic Engineering and Biotechnology (CIGB) Havana, Cuba and mAbs (CMC-H9, CMC-A5) in the Center of Immunology and Biologics (CENIB), Camaguey, Cuba. The inhibition assays were carried out using as reference a commercial equimolar assay for total PSA. For the evaluation, the mAbs were labeled with {sup 125} I by the method of Chloramine T and the capture mAbs were used conjugated with biotin. For separation, it was used the avidin coupled to magnetic particles. To validate the assay we evaluated the sensibility, inter and intraassay precision and the correlation with reference assay in 65 samples of patient under suspicion of prostate cancer. The partner CB-PSA 4 / CB-PSA 9 (capture / tracer) showed the best results in the IRMA. The developed assay presented a detection limit of 0.025 mg/L, a good intra and interassay precision and a high correlation with the reference assay. (author)

  3. Radioiodination of chicken erythrocyte histones H4 and H5 in chromatin. (United States)

    Griffiths, G R; Huang, P C


    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.

  4. 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.


    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.

  5. Oxidative radioiodination damage to human lactoferrin

    Energy Technology Data Exchange (ETDEWEB)

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


    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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Grant, K.I.


    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.

  7. 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


    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)

  8. Antiseptics and microcosm biofilm formation on titanium surfaces

    Directory of Open Access Journals (Sweden)

    Georgia VERARDI


    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.

  9. Novel pyrazole integrated 1,3,4-oxadiazoles: synthesis, characterization and antimicrobial evaluation. (United States)

    Ningaiah, Srikantamurthy; Bhadraiah, Umesha K; Doddaramappa, Shridevi D; Keshavamurthy, Shubakara; Javarasetty, Chethan


    A novel series of 2-(5-methyl-1,3-diphenyl-1H-pyrazol-4-yl)-5-phenyl-1,3,4-oxadiazoles 7(a-m) were synthesized either by cyclization of N'-benzoyl-5-methyl-1,3-diphenyl-1H-pyrazole-4-carbohydrazide 4a using POCl3 at 120°C or by oxidative cyclization of hydrazones derived from various arylaldehyde and (E)-N'-benzylidene-5-methyl-1,3-diphenyl-1H-pyrazole-4-carbohydrazide 5(a-d) using chloramine-T as oxidant. Newly synthesized compounds were characterized by analytical and spectral (IR, (1)H NMR, (13)C NMR and LC-MS) methods. The synthesized compounds were evaluated for their antimicrobial activity and were compared with standard drugs. The compounds demonstrated potent to weak antimicrobial activity. Among the synthesized compounds, compound 7m emerged as an effective antimicrobial agent, while compounds 7d, 7f, 7i and 7l showed good to moderate activity. The minimum inhibitory concentration of the compounds was in the range of 20-50μgmL(-1) against bacteria and 25-55μgmL(-1) against fungi. The title compounds represent a novel class of potent antimicrobial agents.

  10. 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


    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

  11. Synthesis,Crystal Structure and Biological Activity of 2-(Anthracen-9-yl)-5-p-tolyl-1,3,4-oxadiazole

    Institute of Scientific and Technical Information of China (English)

    LI Xin-Wei; HE Dao-Hang


    A novel compound,2-(anthracen-9-yl)-5-p-tolyl-1,3,4-oxadiazole(C23H16N2O),has been synthesized by the condensation of 4-methylbenzohydrazide and anthracene-9-carbaldehyde in an ethanol solution with chloramine-T.The compound was characterized by 1H-NMR,13C-NMR,MS and single-crystal X-ray diffraction.The crystal belongs to the triclinic system,space group P with a = 7.7817(4),b = 8.8544(5),c = 12.4726(8) ,β = 92.8520(10)°,Z = 2,V = 826.58(8) 3,Dc = 1.352 g/cm3,Mr = 336.38,λ(MoKα) = 0.71073 ,μ = 0.084 mm-1,F(000) = 352,R = 0.0381 and wR = 0.1099.The dihedral angle between anthracene skeleton and phenyl ring is 64.19°.A total of 6354 unique reflections were collected,of which 3172 with I 〉 2σ(I) were observed.X-ray analysis indicated an offset face-to-face π-π stacking interaction between anthracene skeletons and an offset face-to-face π-π stacking interaction between phenyl ring planes.The novel compound molecules are connected through the offset face-to-face π-π stacking interactions to generate a three-dimensional network.The preliminary bioassay results showed that the novel compound exhibited significant insect growth inhibitory activity against Spodoptera litura Fabricius larvae.

  12. Mechanistic Investigations of Oxidation of Some Dipeptides by Sodium N-chloro-p-toluenesulfonamide in Alkaline Medium: A Kinetic Study

    Institute of Scientific and Technical Information of China (English)



    The kinetics of oxidation of five dipeptides (DPP) viz., glycylglycine (Gly-Gly), L-alanyl-L-alanine (Ala-Ala),L-valyl-L-valine (Val-Val), L-leucyl-L-leucine (Leu-Leu) and phenylglycyl-phenylglycine (Phg-Phg) by sodium N-chloro-p-toluenesuifonamide or chloramine-T (CAT) in NaOH medium was studied at 308 K. The reactions follow identical kinetics for all the dipeptides, being first-order dependence each on [CAT]o, [DPP]o and fractional-order on [OH-]. Addition of p-toluenesulfonamide or halide ions (CI- or Br-) has no significant effect on the rate of reaction. The reaction rate was found to increase with increase in ionic strength of the medium. The solvent isotope effect was studied using D2O. The activation parameters for the reaction were computed from Arrhenius plots. Equilibrium and decomposition constants were evaluated. The oxidation products of the dipeptides were identiffed as their corresponding aldehydes. An isokinetic relationship was observed with β=352 K, indicating that enthalpy factors control the reaction rate. CH3C6H4SO2NCl- of the oxidant has been postulated as the reactive oxidizing species. Under comparable experimental conditions, the rate of oxidation of the dipeptides increases in the order: Phg-Phg>Ala-Ala>Val-Val>Leu-Leu>Gly-Gly. The kinetics of oxidation of the dipeptides have also been compared with those of their corresponding monomer amino acids. The observed results have been explained by a plausible mechanism and the related rate law has been deduced.

  13. Iodine-123 salmon calcitonin, an imaging agent for calcitonin receptors: synthesis, biodistribution, metabolism and dosimetry in humans

    Energy Technology Data Exchange (ETDEWEB)

    Blower, P.J.; Kettle, A.G.; Leak, A.; O`Doherty, M.J. [Department of Nuclear Medicine, Kent and Canterbury Hospital, Canterbury (United Kingdom); Puncher, M.R.B.; George, S.; Dorsch, S.; Naylor, L.H. [Department of Biosciences, University of Kent, Canterbury (United Kingdom)


    A modified chloramine-T method was used to label a pharmaceutical form of salmon calcitonin (SCT) with iodine-123. Labelling can be performed within 5 min including purification, resulting in >95% radiochemical purity and 70% yield. Digestion analysis shows labelling with two iodine atoms on the tyrosine residue. A Chinese hamster ovary cell-based assay showed that the receptor binding and activation were not impaired by the labelling. Biodistribution in mice was similar to that of commercially available mono-iodinated {sup 125}I-labelled SCT, kidney being the principal target organ. Evaluation in three patients previously diagnosed as having Paget`s disease (injected with 37 MBq [{sup 123}I]diiodotyrosyl{sup 22}-SCT, containing less than 4 IU hormone, imaged dynamically up to 0.5 h and at intervals up to 24 h) shows early uptake in liver, kidney and sites of known Paget`s disease but not in normal bone, and later uptake in thyroid and stomach. Blood clearance was fitted to a biexponential with half-lives of 3.4-7.4 min and 3-34 h. Radiation dosimetry was estimated using MIRDOSE 3. The highest doses (mean mGy/MBq) were to thyroid (6.8 x 10{sup -1}) and kidney (6.0 x 10{sup -2}), with a whole-body dose 3.0 x 10{sup -2}. High performance liquid chromatography analysis revealed that urinary radioactivity was mostly in the form of iodide and diiodotyrosine within minutes of injection, indicating rapid in vivo breakdown.(orig. /MG) (orig.) With 8 figs., 1 tab., 28 refs.

  14. Synthesis of 125I Labeled Estradiol-17-Hemisuccinate and Its Binding Study to Estrogen Receptors Using Scintillation Proximity Assay Method

    Directory of Open Access Journals (Sweden)

    Y. Susilo


    Full Text Available Research was carried out to obtain a selective ligand which strongly bind to estrogen receptors through determination of binding affinity of estradiol-17β-hemisuccinate. Selectivity of these compounds for estrogen receptor was studied using Scintillation Proximity Assay (SPA method. Primary reagents required in the SPA method including radioligand and receptor, the former was obtained by labeling of estradiol-17β-hemisuccinate with 125I, while MCF7 was used as the receptor. The labeling process was performed by indirect method via two-stage reaction. In this procedure, first step was activation of estradiol-17β-hemisuccinate using isobutylchloroformate and tributylamine as a catalist, while labeling of histamine with 125I was carried out using chloramin-T method to produce 125I-histamine. The second stage was conjugation of activated estradiol-17β-hemisuccinate with 125I-histamine. The product of estradiol-17β-hemisuccinate labeled 125I was extracted using toluene. Furtherly, the organic layer was purified by TLC system. Characterization of estradiol-17β-hemisuccinate labeled 125I from this solvent extraction was carried out by determining its radiochemical purity and the result was obtained using paper electrophoresis and TLC were 79.8% and 84.4% respectively. Radiochemical purity could be increased when purification step was repeated using TLC system, the result showed up to 97.8%. Determination of binding affinity by the SPA method was carried out using MCF7 cell lines which express estrogen receptors showed the value of Kd at 7.192 x 10-3 nM and maximum binding at 336.1 nM. This low value of Kd indicated that binding affinity of estradiol-17β-hemisuccinate was high or strongly binds to estrogen recepto

  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


    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. Comparative studies of endotoxin uptake by isolated rat Kupffer and peritoneal cells. (United States)

    Fox, E S; Thomas, P; Broitman, S A


    The process of uptake of endotoxin by cells of the reticuloendothelial system is of current interest. Rabbit peritoneal macrophages have been used to study macrophage-endotoxin interactions and have suggested a receptor-mediated process. It is generally believed that the site of in vivo endotoxin clearance is the liver and that this clearance involves the Kupffer cell population. In the current report, the uptake characteristics of iodine-125-labeled Salmonella minnesota lipopolysaccharide (LPS) were compared in both isolated rat Kupffer cells and elicited rat peritoneal cells. Both types of cells were isolated from male Sprague-Dawley rats fed a semisynthetic AIN-76 5% saturated-fat diet either by peritoneal lavage for peritoneal cells or by collagenase perfusion followed by purification on a 17.5% metrizamide gradient for Kupffer cells. Hot phenol water-extracted S. minnesota LPS was labeled with iodine by the chloramine-T method following a reaction with methyl-p-hydroxybenzimidate. The in vitro uptake of [125I]LPS by Kupffer cells was unsaturable up to concentrations of 33.33 micrograms/ml, while peritoneal cells became saturated at between 16.67 and 25 micrograms of LPS per ml. Uptake by both types of cells could be inhibited by a 10-fold excess of unlabeled LPS. Kinetic experiments demonstrated that Kupffer cells were unsaturable after 60 min of incubation, while peritoneal cells were saturable after 40 min of incubation. Pretreatment with 75 mM colchicine inhibited uptake by peritoneal cells but not Kupffer cells, while pretreatment with 12 mM 2-deoxyglucose inhibited uptake by Kupffer cells but not peritoneal cells. These results are consistent with a process of receptor-mediated endocytosis for peritoneal cells, while Kupffer cells may internalize endotoxins by absorptive pinocytosis. These results suggest that studies of peritoneal cell-endotoxin interactions do not accurately describe the physiologic process within the liver, the major site for the

  17. 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.


    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.

  18. Excretory-secretory and somatic antigens in the diagnosis of human filariasis. (United States)

    Kaushal, N A; Hussain, R; Ottesen, E A


    In order to compare the immunodiagnostic value of excretory-secretory (E-S) antigens derived from adult Brugia malayi worms with somatic antigens derived from adults, microfilariae (Mf) and infective larvae (L3) of these parasites, well defined serum pools from patients with filarial (brugia, bancrofti, loa and perstans) and non-filarial (ascaris, stronglyoides, toxocara, echinococcus, cysticercus and schistosoma) helminth infections were tested against antigens derived from these different life cycle stages of B. malayi in a Staphylococcus aureus radioimmunoprecipitation assay (S. aureus RIA). The adult brugia antigens proved significantly more discriminatory than those of the other parasite stages, with the homologous brugia serum pool also showing greater reactivity to adult than to L3 and Mf antigens. Similar results were obtained when individual sera from patients (rather than serum pools) were tested in the same assay. The most surprising finding was the minimal reactivity seen between the adult filarial antigens and the non-filarial serum pools despite the presence in these pools of strong antibody reactivity with their homologous antigens. The reasons underlying the unexpected specificity of this S. aureus RIA for discriminating among sera from filarial and non-filarial infections were analysed qualitatively by immunoprecipitation techniques. It was found that use of the chloramine-T method for radioiodination resulted in preferential labelling of the low molecular weight (mol. wt) proteins (10-70,000 daltons) in the B. malayi adult somatic antigen and that these antigens were bound primarily by the filarial and not the non-filarial serum pools. These findings suggest that lower mol. wt helminth antigens may show greater species specificity than those with higher mol. wt, and those with higher mol. wt, greater cross-reactivity. If substantiated by further analysis, such results would have important implications for the subsequent isolation of diagnostically

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

    Energy Technology Data Exchange (ETDEWEB)

    Teitelbaum, Anne P.


    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 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.

  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)


    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. Iodine-125 metaraminol: A new platelet specific labeling agent

    Energy Technology Data Exchange (ETDEWEB)

    Ohmomo, Y.; Yokoyama, A.; Kawaii, K.; Horiuchi, K.; Saji, H.; Torizuka, K.


    In the search for a platelet specific labeling agent, Metaraminol (MA), which is a sympatomimetic amine used for the treatment of hypotension, cardiogenic shock and well recognized as a drug actively incorporated and accumulated in platelet, attracted the authors' attention. Using the classical chloramine-T iodination method, a high labeling efficiency near 98%, reaching a specific activity up to about 1000 Ci/mmole was obtained. Upon the harvest of platelet, only as platelet rich plasma (PRP), the labeling with this radiopharmaceutical was easily performed by incubation at 37/sup 0/C for 10 min. Labeling efficiency as high as 63.0 +- 3.1% at 24 x 10/sup 8/ cells/ml was obtained. In in-vitro studies, the unaltered state of I-125 MA labeled platelet, with their cellular functions fully retained was demonstrated. Pharmacological study indicated a specific incorporation of I-125 MA by active transport system similar to that of 5-HT, along with passive diffusion. Then the in-vivo study carried out in rabbits with induced thrombi on the femoral artery, showed rather rapid disappearance of the I-125 MA labeled autologous platelet radioactivity, from circulating blood reaching as high thrombus-to-blood activity ratio as 19.8+-4.3 within 30 min post-administration. This new platelet labeling agent, I-125 MA, has many advantages over the use of IN-111 oxine and holds considerable promise for thrombus imaging with single photon emission CT upon the availability of I-123 MA.

  2. (/sup 125/I) radioiodinated metaraminol: A new platelet-specific labeling agent

    Energy Technology Data Exchange (ETDEWEB)

    Ohmomo, Y.; Yokoyama, A.; Kawai, K.; Arano, Y.; Horiuchi, K.; Tanaka, C.; Saji, H.; Torizuka, K.


    In our search for a platelet-specific labeling agent, metaraminol (MA), a low-toxic pharmaceutical for the treatment of hypotension and cardiogenic shock, attracted our attention. Its active incorporation and accumulation by platelets have been recognized. At first, the preparation of /sup 125/I radioiodinated metaraminol (/sup 125/I-MA) was carried out using the chloramine-T method. Then, upon the harvest of platelets as platelet-rich plasma (PRP), their labeling with this new radiopharmaceutical was easily performed by incubation for 10 min at 37/sup 0/C. The cell-labeling efficiency was dependent on cell density, reaching 63.0%+-3.1% at 2.4x10/sup 9/ cells/ml. The specific incorporation of /sup 125/I-MA by an active transport system similar to that of 5-hydroxytryptamine (5-HT) as well as by passive diffusion was demonstrated. In vitro studies, the unaltered state of /sup 125/I-MA-labeled platelets with their cellular functions fully retained was estimated. In vivo studies carried out in rabbits with induced thrombi in the femoral artery showed a rather rapid disappearance of the radioactivity from circulating blood, reaching a high thrombus-to-blood activity ratio of 19.8+-4.3 within 30 min of the administration of /sup 125/I-MA-labeled autologous platelets. Thus, with the potential availability of /sup 123/I, /sup 123/I-MA-labeled platelets appear to be a promising agent for thrombus imaging using single-emission computed tomography (CT) studies.

  3. N-Chlorotaurine and ammonium chloride: an antiseptic preparation with strong bactericidal activity. (United States)

    Gottardi, Waldemar; Arnitz, Roland; Nagl, Markus


    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.

  4. Radioimmunoimaging of osteogenic sarcoma xenografts in nude mice using monoclonal antibodies to osteogenic sarcoma

    Energy Technology Data Exchange (ETDEWEB)

    Sakahara, H.; Endo, K.; Nakashima, T.; Koizumi, M.; Ohta, H.; Kunimatsu, M.; Torizuka, K.; Nakamura, T.; Tanaka, H.; Kotoura, Y.


    The authors have developed several monoclonal antibodies against human osteogenic sarcoma, one of which; OST7 (IgGl) selectively localized in osteogenic sarcoma xenografts in nude mice. In the present study, F(ab')/sub 2/ fragment was compared with whole IgG and those labeled with In-111 as well as I-131 were used as a radiotracer for the scintigraphic imaging of tumors. IgC and F(ab')/sub 2/ were labeled with I-131 using chloramine-T method and injected into nude mice bearing human osteogenic sarcoma. Scintigrams at day 2 clearly delineated the site of tumors with almost no radioactivity in other organs with F(ab')/sub 2/, which yielded much better images than whole IgG. Tumor-to-blood ratio of 6.09-27.87 was obtained at day 2 using F(ab')/sub 2/, whereas it was 0.76-1.12 at day 2 and 2.05-3.27 at day 7 with IgG. I-131 labeled nonspecific F(ab')/sub 2/ or IgG resulted in no or very low tumor uptake with tumor-to-blood ratio of 0.94-1.18 at day 2 for F(ab')/sub 2/ and 0.67-0.76 at day 7 for IgG, respectively. In-111 labeled F(ab')/sub 2/ fragment of OST7, which was prepared using DTPA as a bifunctional chelate, also showed a high tumor accumulation with tumor-to-blood ratio of 11.67-17.54 at day 2, but higher background activity in the liver and kidney was observed than I-131 labeled one. These results indicate that F(ab')/sub 2/ fragment of OST7 labeled with either I-131 or In-111, has a great potential for the radioimmunoimaging of osteogenic sarcoma.

  5. Simple spectrophotometric and titrimetric methods for the determination of sulfur dioxide. (United States)

    Yogendra Kumar, M S; Gowtham, M D; Mahadevaiah; Agendrappa, G


    The proposed work describes a simple spectrophotmetric as well as a titrimetric method to determine sulfur dioxide. The spectrophotometric method is based on a redox reaction between sulfur dioxide and iodine monochloride obtained from iodine with chloramine-T in acetic acid. The reagent iodine monochloride oxidizes sulfur dioxide to sulfate, thereby reducing itself to iodine. Thus liberated iodine will also oxidize sulfur dioxide and reduce itself to iodide. The obtained iodide is expected to combine with iodine to form a brown-colored homoatomictriiodide anion (460 nm), which forms an ion-pair with the sulfonamide cation, providing exceptional color stability to the system under an acidic condition, and is quantitatively relatd to sulfur dioxide. The system obeys Beer's law in the range 5 - 100 microg of sulfur dioxide in a final volume of 10 ml. The molar absorptivity is 5.03 x 10(3) l mol(-1)cm(-1), with a relative standard deviation of 3.2% for 50 microg of sulfur dioxide (n = 10). In the titrimetric method, the reagent iodine monochloride was reduced with potassium iodide (10%) to iodine, which oxidized sulfur dioxide to sulfate, and excess iodine was determined with a thiosulfate solution. The volume difference of thiosulfate with the reagent and with the sulfur dioxide determined the sulfur dioxide. Reproducible and accurate results were obtained in the range of 0.1 - 1.5 mg of sulfur dioxide with a relative standard deviation of 1.2% for 0.8 mg of sulfur dioxide (n = 10).

  6. 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


    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.

  7. Biodistribution and tumor uptake of C60(OH) x in mice (United States)

    Ji, Zhi Qiang; Sun, Hongfang; Wang, Haifang; Xie, Qunying; Liu, Yuangfang; Wang, Zheng


    Radiolabeling of fullerol, 125I-C60(OH) x , was performed by the traditional chloramine-T method. The C-I covalent bond in I-C60(OH) x was characterized by X-ray photoelectron spectroscopy (XPS) that was sufficiently stable for in vivo study. Laser light scattering spectroscopy clearly showed that C60(OH) x aggregated to large nanoparticle clumps with a wide range of distribution. The clumps formed were also visualized by transmission electron microscope (TEM). We examined the biodistribution and tumor uptake of C60(OH) x in five mouse bearing tumor models, including mouse H22 hepatocarcinoma, human lung giantcellcarcinoma PD, human colon cancer HCT-8, human gastric cancer MGC803, and human OS732 osteosarcoma. The accumulation ratios of 125I-C60(OH) x in mouse H22 hepatocarcinoma to that in normal muscle tissue (T/N) and blood (T/B) at 1, 6, 24 and 72 h, reveal that 125I-C60(OH) x gradually accumulates in H22 tumor, and retains for a quite long period (e.g., T/N 3.41, T/B 3.94 at 24 h). For the other four tumor models, the T/N ratio at 24 h ranges within 1.21-6.26, while the T/B ratio ranges between 1.23 and 4.73. The accumulation of C60(OH) x in tumor is mostly due to the enhanced permeability and retention effect (EPR) and the phagocytosis of mononuclear phagocytes. Hence, C60(OH) x might serve as a photosensitizer in the photodynamic therapy of some kinds of tumor.

  8. Biodistribution and tumor uptake of C{sub 60}(OH){sub x} in mice

    Energy Technology Data Exchange (ETDEWEB)

    Ji Zhiqiang; Sun Hongfang, E-mail:; Wang Haifang; Xie Qunying; Liu Yuangfang [Peking University, Department of Chemical Biology, College of Chemistry and Molecular Engineering (China); Wang Zheng [Chinese Academy of Medical Sciences, Cancer Institute (China)


    Radiolabeling of fullerol, {sup 125}I-C{sub 60}(OH){sub x}, was performed by the traditional chloramine-T method. The C-I covalent bond in I-C{sub 60}(OH){sub x} was characterized by X-ray photoelectron spectroscopy (XPS) that was sufficiently stable for in vivo study. Laser light scattering spectroscopy clearly showed that C{sub 60}(OH){sub x} aggregated to large nanoparticle clumps with a wide range of distribution. The clumps formed were also visualized by transmission electron microscope (TEM). We examined the biodistribution and tumor uptake of C{sub 60}(OH){sub x} in five mouse bearing tumor models, including mouse H22 hepatocarcinoma, human lung giantcellcarcinoma PD, human colon cancer HCT-8, human gastric cancer MGC803, and human OS732 osteosarcoma. The accumulation ratios of {sup 125}I-C{sub 60}(OH){sub x} in mouse H22 hepatocarcinoma to that in normal muscle tissue (T/N) and blood (T/B) at 1, 6, 24 and 72 h, reveal that {sup 125}I-C{sub 60}(OH){sub x} gradually accumulates in H22 tumor, and retains for a quite long period (e.g., T/N 3.41, T/B 3.94 at 24 h). For the other four tumor models, the T/N ratio at 24 h ranges within 1.21-6.26, while the T/B ratio ranges between 1.23 and 4.73. The accumulation of C{sub 60}(OH){sub x} in tumor is mostly due to the enhanced permeability and retention effect (EPR) and the phagocytosis of mononuclear phagocytes. Hence, C{sub 60}(OH){sub x} might serve as a photosensitizer in the photodynamic therapy of some kinds of tumor.

  9. [A sensitive and specific radioimmunoassay for arginine vasopressin and its validation]. (United States)

    Oki, Y; Ohgo, S; Yoshimi, T


    A sensitive and specific radioimmunoassay (RIA) for arginine vasopressin (AVP) has been developed and validated. Synthetic AVP was coupled to bovine serum albumin (BSA) with glutaraldehyde. Antisera against AVP were raised in three rabbits immunized with AVP-BSA complex. After 6 months, at the 16th injection, one of the antisera had a titer high enough to be utilizable for RIA at a final dilution of 1:400,000. The labeling of AVP with 125I Na was performed with the modified chloramine T method, and the purification of iodinated AVP was done with gel filtration chromatography on a Sephadex G-25 fine column (1 X 20 cm) with an elution buffer of 0.01 M acetic acid containing 0.1% BSA. Radioactivities from the Sephadex G-25 were eluted in three peaks. 125I-AVP, which was reactive to the antiserum, was contained in the third peak, and 125I-AVP in the fractions on the down slope of the peak was used for the radioligand in the amount of 1000 cpm. The specific activity of purified 125I-AVP was about 400 muCi/microgram. Diluted antiserum and samples, unlabeled AVP or related peptides were preincubated at 4 degrees C for 24 hr, and then 125I-AVP was added to the mixture and incubated for a further 72 hr. Separation of B and F was done with polyethyleneglycol. The minimal detection limit of AVP, which was 95% of the confidence limit of the mean value of B0, was 0.4 pg/tube. The cross-reactivities with lysine vasopressin, arginine vasotocin, DDAVP and oxytocin were 0.1%, 30%, 1% and 0%, respectively. AVP in plasma was extracted with cold acetone and petroleum ether. The recoveries of synthetic AVP from plasma which was added (2-16 pg) were more than 94%. The intra and inter-assay coefficients of variation determined by plasma of AVP concentration of about 4.8 pg/ml were 8.7% and 11.3%, respectively. The RIA detected AVP of concentration as low as 1 pg/ml following the extraction procedure. AVP immunoreactivity was detected without extraction in urine, and the lyophilized

  10. Spectroscopic analysis of the impact of oxidative stress on the structure of human serum albumin (HSA) in terms of its binding properties (United States)

    Maciążek-Jurczyk, M.; Sułkowska, A.


    Oxygen metabolism has an important role in the pathogenesis of rheumatoid arthritis (RA). Reactive oxygen species (ROS) are produced in the course of cellular oxidative phosphorylation and by activated phagocytic cells during oxidative bursts, exceed the physiological buffering capacity and result in oxidative stress. ROS result in oxidation of serum albumin, which causes a number of structural changes in the spatial structure, may influence the binding and cause significant drug interactions, particularly in polytherapy. During the oxidation modification of amino acid residues, particularly cysteine and methionine may occur. The aim of the study was to investigate the influence of oxidative stress on human serum albumin (HSA) structure and evaluate of possible alterations in the binding of the drug to oxidized human serum albumin (oHSA). HSA was oxidized by a chloramine-T (CT). CT reacts rapidly with sulfhydryl groups and at pH 7.4 the reaction was monitored by spectroscopic techniques. Modification of free thiol group in the Cys residue in HSA was quantitatively determined by the use of Ellman's reagent. Changes of albumin conformation were examined by comparison of modified (oHSA) and nonmodified human serum albumin (HSA) absorption spectra, emission spectra, red-edge shift (REES) and synchronous spectroscopy. Studies of absorption spectra indicated that changes in the value of absorbance associated with spectral changes in the region of 200-250 nm involve structural alterations in peptide backbone conformation. Synchronous fluorescence spectroscopy technique confirmed changes of position of tryptophanyl and tyrosyl residues fluorescent band caused by CT. Moreover analysis of REES effect allowed to observe structural changes caused by CT in the region of the hydrophobic pocket containing the tryptophanyl residue. Effect of oxidative stress on binding of anti-rheumatic drugs, sulfasalazine (SSZ) and sulindac (SLD) in the high and low affinity binding sites was

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

    Directory of Open Access Journals (Sweden)

    Benevolo Maria


    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

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

    Directory of Open Access Journals (Sweden)

    C.M.L. Vasconcelos


    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

  13. 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)


    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

  14. 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)


    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

  15. 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


    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

  16. 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))


    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.

  17. 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


    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

  18. 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


    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.

  19. Assembling of ({beta}LPH) beta-lypothrophine radioimmunoassay. Plasma levels standardization in normal individuals and patients with hypophysis and adrenals diseases; Montagem do radioimunoensaio da beta-lipotrofina ({beta}LPH). Padronizacao dos niveis plasmaticos em individuos normais e em pacientes com doencas hipofisarias e adrenais

    Energy Technology Data Exchange (ETDEWEB)

    Castro, Margaret de


    The present study investigates the extraction and radioimmunoassay (RIA) conditions of plasma {beta}LPH. It was extracted by the activated silicic acid method, with a mean extraction efficiency of 31.6% and a mean intra-extraction variation coefficient of 8.1%. Radioiodination was performed by the chloramine-T method and {beta}LPH {sup 125} I was purified by gel chromatography on Sephadex G100. Estimated specific activity ranged from 100 to 192.8 {mu}Ci/{mu}g, with a mean incorporation percentage of 66.6%. The titer of the first antibody was 1:50.000/100 {mu}l. The assay was performed under non-equilibrium conditions, with a pre-incubation period of 24 hours and incubation of 4 hours. Mean immunoreactivity (Bo/Total) was 21.1%, with a mean Blank/Total ratio of 2.3%. Sensitivity, expressed as the mean minimum detectable dose, was 40 pg/tube, equivalent to 56 pg/ml plasma. Intra-assay variation coefficients were 6.5%, 3.8% and 6.8%, respectively, at B/Bo levels of 0.8, 0.6 and 0.4 of the standard curve. At B/Bo equal to 0.5, the intra-assay variation coefficient was 20.9%. Replicates of 14 plasma samples showed a correlation coefficient of r 0.99, (p< 0.05). Parallelism between the curve obtained with different volumes of an extract with a high {beta}LPH value and the standard curve was found. The method was controlled biologically by the presence of correlation between the plasma {beta}LPH levels and determined pathological states and with clinical functional studies. Twenty seven normal individuals, 10 patients with Cushing`s disease to a tumor of the hypophysis, 4 patients with Cushing syndrome due to an adrenal tumor, 10 patients Addison disease, and 8 patients with hypopituitarism were studied. (author). 119 refs., 28 figs., 2 tabs.

  20. Advanced oxidation protein products in plasma: stability during storage and correlation with other clinical characteristics. (United States)

    Matteucci, E; Biasci, E; Giampietro, O


    Proteins are susceptible to free radical damage. We measured advanced oxidation protein products (AOPP) in the plasma of 56 hospitalised patients. Concentrations of AOPP were expressed as chloramine-T equivalents by measuring absorbance in acidic conditions at 340 nm in the presence of potassium iodide. We also determined erythrocyte sedimentation rate (ESR), circulating urea, creatinine, glucose, uric acid, electrolytes, lipids, total proteins and fractions and fibrinogen. Twenty-four samples were processed both immediately and after 7, 15, 30, 90, 180 and 438 days of storage at both at -20 degrees C and -80 degrees C (aliquots were frozen and thawed only once) to evaluate AOPP stability. The remaining 32 samples were also processed for thiobarbituric-acid-reactive substances (TBARS). Mean AOPP concentration in all 56 patients was 48.3+/-37.2 microM. Mean basal concentration of AOPP in the 24 plasma samples (55.0+/-47.1 microM) showed no significant change at each intermediate determination, yet significantly increased after 438 days of storage both at -80 degrees C (96.6+/-83.2, p<0.01) and, markedly, at -20 degrees C (171.3+/-94.6, p<0.001). TBARS concentration was 1.59+/-0.65 micromol/l. Multiple regression analysis evidenced that AOPP concentration was positively correlated (multiple r=0.62, p<0.001) with serum urea and triglycerides, but negatively correlated with patient age (indeed, serum albumin and total proteins decreased with increasing age, r=0.3, p<0.05). TBARS concentration was associated with ESR and serum glucose (multiple r=0.73, p<0.001), yet positively with AOPP (r=0.39, simple p<0.05). We conclude that AOPP remain stable during sample storage both at -20 degrees C and -80 degrees C for 6 months. Renal failure and hypertriglyceridemia probably enhance the in vivo process of AOPP formation. Oxidative damage as measured by TBARS may be increased because of exposure to hyperglycemia causing nonenzymatic glycation of plasma proteins.

  1. Research on the Technology of Exterminating the Defacing Organisms Limnoperna Fortunei by Oxidant in Water Delivery Systems%输水系统中污损生物沼蛤的氧化剂灭杀技术研究

    Institute of Scientific and Technical Information of China (English)

    魏小熙; 杨正健; 刘德富; 谭纤茹


    近年来,我国南方输水工程常受沼蛤(limnoperna fortunei)生物污损的危害。沼蛤是一种污损性入侵生物,入侵输水工程后聚团附着于输水系统表面可腐蚀附着面、降低管道过流能力、污染水质等。以沼蛤防治为目的研究二氧化氯(ClO2)、氯胺T(C7H7ClNNaO2S·3(H2O))、高锰酸钾(KMnO4)、次氯酸钠(NaClO)和过氧化氢(H2O2)5种水处理常用氧化剂对沼蛤的杀灭效果及足丝溶解特性,以得出基于化学氧化剂的沼蛤防治措施。结果表明:次氯酸钠为5种氧化剂中灭杀沼蛤的最佳试剂,死亡率随次氯酸钠投加浓度增加而增大,但次氯酸钠投加浓度大于20 mg/L时水体中pH等环境条件受到较大影响,依据水体中有效氯衰减规律及灭杀效果,次氯酸钠作为沼蛤杀灭剂初始投加的有效氯浓度宜为10.0~20.0 m g/L。%In recent years ,water project in South China is often affected by the defacing hazards of limnoperna fortunei ,which is a kind of defacement of invasive species .Limnoperna fortunei attaches together to the surface of the water delivery system when they invade the water conveyance project ,which can corrode the adhesion surface ,reduce the capacity of pipeline flow ,pollute the water etc .For the purpose of prevention of limnoperna fortunei ,we study the chlorine dioxide (ClO2 ) ,chloramine-T(C7 H7ClNNaO2S · 3(H2O),potassiumpermanganate(KMnO4),sodiumhypochlorite(NaClO)andhydrogenperoxide(H2O2),thefivekindsofcom‐monly used water treatment oxidant on the elimination effect and byssus dissolution characteristics of limnoperna fortunei ,in order to get its prevention and control measures based on the chemical oxidants .From the experiment we can get the following conclusions . Sodium hypochlorite is the best reagent of the five oxidants to kill limnoperna fortunei ,and with the increase in the initial active chlo‐rine concentration ,the mortality rate of


    Institute of Scientific and Technical Information of China (English)

    刘真; 张林; 熊鸿燕


    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.

  3. Utilization of N-X bonds in the synthesis of N-heterocycles. (United States)

    Minakata, Satoshi


    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

  4. 门静脉灌注胶原酶逆转四氯化碳致实验兔肝硬化%Portal administration of collagenase reverse liver cirrhosis in rabbit CCl4 model

    Institute of Scientific and Technical Information of China (English)

    邱慧彬; 金博; 孙涛; 于晓红; 杨英祥; 付山峰; 路平


    目的 观察门静脉灌注胶原酶对逆转兔实验性肝硬化的作用。方法 实验动物分4组,1组:四氯化碳注射12周后行门静脉置管,1周后开始门静脉灌注胶原酶4周;2组:四氯化碳注射12周后行门静脉置管,1周后开始门静脉灌注生理盐水4周;3组:橄榄油注射12周后行门静脉置管,1周后开始门静脉灌注胶原酶4周;4组:橄榄油注射12周后行门静脉置管,1周后开始门静脉灌注生理盐水4周。以上各组取肝组织,用氯胺T法检测门静脉灌注实验前后肝组织的羟脯氨酸含量及肝组织的病理检查。结果 经过的四氯化碳(CCL4)注射和胶原酶门静脉灌注后,应用胶原酶灌注的动物肝羟脯氨酸含量显著低于用生理盐水灌注的对照组,肝组织学检查也显示胶原酶治疗后肝硬化消退,肝肾功能检查未发现胶原酶的毒性反应。结论 门静脉灌注胶原酶可以促进已形成的肝硬化向正常肝组织结构逆转,并未发现对肝、肾组织毒性反应。%Objective To investigate whether portal collagenase administration can reverse liver cirrhosis. Methods Rabbits divided into 4 groups; group 1: Rabbits received carbon tetrachloride (CC14) subcutaneously (sc) for 12 weeks,then have an operation to put an cannulation in portal vein, collagenase portally for 4 weeks, group 2: Rabbits received CC14 sc for 12 weeks, then have an operation to put an cannulation in portal vein, normal saline portally for 4 weeks, group 3: Rabbits received olive oil sc for 12 weeks,then have an operation to put an cannulation in portal vein collagenase portally for 4 weeks, group 4: Rabbits received olive oil sc for 12 weeks,then have an operation to put an cannulation in portal vein normal saline portally for 4 weeks. Then take tissues of liver, portal vein, brain,kidney, lung and heart in 4 groups. After the zoopery,use chloramine T to test the hydroxypro-line content in liver tissues after

  5. New Methods for Labeling RGD Peptides with Bromine-76

    Directory of Open Access Journals (Sweden)

    Lixin Lang, Weihua Li, Hong-Mei Jia, De-Cai Fang, Shushu Zhang, Xilin Sun, Lei Zhu, Ying Ma, Baozhong Shen, Dale O. Kiesewetter, Gang Niu, Xiaoyuan Chen


    Full Text Available Direct bromination of the tyrosine residues of peptides and antibodies with bromine-76, to create probes for PET imaging, has been reported. For peptides that do not contain tyrosine residues, however, a prosthetic group is required to achieve labeling via conjugation to other functional groups such as terminal α-amines or lysine ε-amines. The goal of this study was to develop new strategies for labeling small peptides with Br-76 using either a direct labeling method or a prosthetic group, depending on the available functional group on the peptides. A new labeling agent, N-succinimidyl-3-[76Br]bromo-2,6-dimethoxybenzoate ([76Br]SBDMB was prepared for cyclic RGD peptide labeling. N-succinimidyl-2, 6-dimethoxybenzoate was also used to pre-attach a 2, 6-dimethoxybenzoyl (DMB moiety to the peptide, which could then be labeled with Br-76. A competitive cell binding assay was performed to determine the binding affinity of the brominated peptides. PET imaging of U87MG human glioblastoma xenografted mice was performed using [76Br]-BrE[c(RGDyK]2 and [76Br]-BrDMB-E[c(RGDyK]2. An ex vivo biodistribution assay was performed to confirm PET quantification. The mechanisms of bromination reaction between DMB-c(RGDyK and the brominating agent CH3COOBr were investigated with the SCRF-B3LYP/6-31G* method with the Gaussian 09 program package. The yield for direct labeling of c(RGDyK and E[c(RGDyK]2 using chloramine-T and peracetic acid at ambient temperature was greater than 50%. The yield for [76Br]SBDMB was over 60% using peracetic acid. The conjugation yields for labeling c(RGDfK and c(RGDyK were over 70% using the prosthetic group at room temperature. Labeling yield for pre-conjugated peptides was over 60%. SDMB conjugation and bromination did not affect the binding affinity of the peptides with integrin receptors. Both [76Br]Br-E[c(RGDyK]2 and [76Br]BrDMB-E[c(RGDyK]2 showed high tumor uptake in U87MG tumor bearing mice. The specificity of the imaging tracers

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

    Directory of Open Access Journals (Sweden)

    Maria Tereza Colturato


    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

  7. 平阳霉素-活性炭纳米微粒行口腔癌淋巴化疗的靶向性评价%Lymphatic targeting study of pingyangmycin-activated carbon nanoparticles treating oral cancer lymph node metastasis

    Institute of Scientific and Technical Information of China (English)

    孙明磊; 谢卫红; 王昌美; 温玉明


    目的 观察平阳霉素-活性炭纳米微粒(PYM-CH-NP)经鼠癌周黏膜下给药后,活性药物在体内各组织器官中的分布情况,探讨PYM-CH-NP对淋巴结转移灶的靶向性.方法 采用改良氯胺T法将125I标记平阳霉素(PYM),应用淋巴结高转移癌株U14建立昆明小鼠颈淋巴结转移模型.将360只颈淋巴结转移模型小鼠随机分为PYM-CH-NP组、PYM组和阴性对照组,分别于癌周黏膜下注射PYM-CH-NP、PYM水溶液和空白活性炭纳米微粒,给药后0.5、1、4、8、12、24、48、72、96、120、144、168 h处死动物,取血、心、肝、脾、肺、肾和颈淋巴结,检测各时间点各组织器官内平阳霉素的放射活性,计算各样本单位重量的药物放射活性(比放射活性),并计算药物的选择性指数(SI)和靶向指数(TI).结果 PYM-CH-NP组各时间点颈淋巴结内药物浓度显著高于PYM组(P<0.001),4 h后血、心、肝、脾、肺、肾等器官中药物浓度均低于PYM组(P<0.001).PYM组各时间点的SI均小于1,而PYM-CH-NP组SI和TI最低值为1.793和1.562,最高为72 h时的68.126和14.623.结论 PYM-CH-NP可显著增加淋巴结转移灶内的药物浓度,同时非靶器官药物分布减少,降低了药物的全身毒副反应.%Objective To investigate the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin-activated carbon nanoparticles (PYM-CH-NP) and evaluate the lymph targeting effect of PYM-CH-NP. Methods Pingyangmycin (PYM) was radiolabeled with 125I by modified the chloramine T method. Cervical lymph node metastasis mice model was established by buccal submucosa inoculation of a high lymph metastasis cell line U14 cancer cell. 360 mice models burdened with cervical lymph metastasis were randomly divided into 3 groups. PYM group was treated with PYM water solution, PYM-CH-NP group was treated with PYM-CH-NP. Negative control group was injected with activated carbon

  8. 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


    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

  9. Effects of “two fairy decoction”(二仙汤) and its decomposed recipe on gonadotropin-releasing hormone secreted by GT1-7 cell line%二仙汤及其拆方对GT1-7细胞株GnRH分泌的影响

    Institute of Scientific and Technical Information of China (English)

    杨颖; 陈名道; 李凤英; 唐金凤; 高国锋; 陈家伦


    目的:GT1-7细胞株是转入了猴病毒-40的T抗原(SV40T)癌基因的促性腺激素释放激素(GnRH)神经内分泌细胞株。本实验观察补肾方二仙汤及其“温肾”、“滋阴”两个拆方对GT1-7细胞株释放GnRH的影响。方法:(1)3个月龄的SD雄性大鼠予成人每公斤体重的10倍剂量灌服中药4日,于末次给药后1或2小时腹主动脉取血,制备成药物血清。(2)氯胺T法标记GnRH,建立稳定可靠的放免标准曲线。(3)GT1-7细胞用含10%、30%或50%药物血清的培养液孵育24或48小时,收集上清液做放射免疫测定。结果:(1)末次给药后1、2小时取血的二仙汤药物血清均能刺激GnRH释放,以1小时给药血清效果最好。(2)10%浓度的药物血清为最有效剂量。(3)二仙汤全方及其两个拆方均能促进GnRH释放,以全方效果最显著。结论:中医“肾主生殖”的功能似涉及下丘脑GnRH神经元及其调控性腺轴的功能,二仙汤及其全方能够直接调节GnRH的分泌。%To investigate the effects of kidney tonifying formula “two fairy decoction”(TFD,二仙汤) and its two decomposed recipes “kidney warming”(温肾) and “yin nourishing”(滋阴) of ingredients on GT1-7 cell line which was developed by targeting the SV40T antigen to gonadotropin-releasing hormone(GnRH) neuron in transgenic mouse.Methods:(1)Three months-aged male rats were orally administered herbal extract for four days,sera were obtained from rats one or two hours after the last administration.(2)GnRH was measured by radioimmunoassay(RIA) with chloramine T iodination,and the detectable limit was 0.25 pg.(3)Cells were maintained for 24 or 48 hours with DMEM which contained 10%,30% and 50% rat serum,and samples were collected for RIA measurements.Results:(1)Both sera obtained from one and two hours after the last oral herb administration of TFD stimulated GnRH secretion,the former seemed more potent.(2

  10. 玷污层及不同根尖倒充填材料对根尖倒充填后微渗漏影响的体外研究%In vitro studies of smear layer and retrograde fil ing material impact on apical microleakage after down -fil ed

    Institute of Scientific and Technical Information of China (English)

    李永立; 张丽萍; 武燃; 刘鹏


    Objective:Af ect apical microleakage after fil ing material with the smear layer on the application of three root canal.Method:Select 64 single root canal teeth after root canal preparation,cold lateral condensation gut a -percha root canal fil ing,after removal of the apical root canal preparation down.Randomly divided into experimental group and control group (n =30 each).Experimental group:17%EDTA +1% chloramine -T remove the smear layer,In the control group:5ml saline.Randomly selected from two groups do SEMobservation.Each group was randomly divided into 3 groups were MTA,GIC,HG were down fil ing,taking root canal fil ing the upper section.Placing micro -leakage model.First detected in concentrations ranging from 0 days 1,2,4,7,10,15,20,30 crown direction leaked square root of glucose.Results:MTA's closure at each time point were superior GIC,HG.Dif erent back molars same treatment (re-moval or retention)under the fil ing material compared pairwise dif erences were statistical y significant P 0.05).Conclusion:MTA apical closure than GIC,HG,application MTA,GIC as a retrograde fil ing material after removal of the smear lay-er apical sealing bet er,but no dif erence in HG.%目的:探讨对应用3种根管倒充填材料玷污层去留对根尖微渗漏的影响,以期为临床应用提供依据。方法:选取64颗单根管牙,常规根管预备后,冷牙胶侧向加压充填根管,根尖切除后进行根管倒预备,将其随机分为实验组与对照组(每组各32颗)。实验组:17%EDTA +2%氯亚明去除玷污层,对照组:用5mL 生理盐水冲洗。从2组中随机抽取 2颗牙做扫面电镜检测。各组分别用 MTA、GIC 和 HG 进行倒充填。去除根管上段充填物,将离体牙置于微渗漏模型,于第1、2、4、7、10、15、20和30天检测从冠方向根方漏出的葡萄糖的浓度。结果:MTA 的封闭性在各个时间点均优于 GIC、HG。离体牙同一种处理方法(去除或保留

  11. 抗ProGRP(31-98)单克隆抗体E-B5的131I标记及体内生物分布研究%The in vivo pharmacokinetics study of anti-ProGRP(31-98) monoclonal antibody E-B5

    Institute of Scientific and Technical Information of China (English)

    徐巧玲; 周小林; 石怡珍; 杨君; 刘增礼


    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 af-ter injection of 131I-E-B5 antibody. Continuous variables were expressed as x±s and compared by t-test with SPSS 13.0 software. Results The labeling efficiency and radiochemical purity of131I-E-B5were 90.8%, 99.28%, respectively. The radiochemical purity still maintained above 70% after incubation in water bath at 37℃ 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 respective-ly. 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, t1/2α= 8.35 h. The metabolism of 131I-E-B5 antibody mainly depended on liver and kidney and with rapid elimination in blood. Condusions 131I-E-B5 antibody not only has high la-beling efficiency and radioehemical purity, but also has good stability and keeps good immunological activi-ty. 131I-E-B5 is a promising agent of radioimmunoimaging and radioimmunotherapy of SCLC.%目的 研究抗胃泌素释

  12. 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


    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

  13. Preparation and lymphatic targeting study of pingyangmycin-activated charcoal nanoparticles%平阳霉素纳米活性炭的研制及其淋巴靶向性研究

    Institute of Scientific and Technical Information of China (English)

    孙明磊; 尚君兰; 王海斌; 王昌美; 温玉明


    Objective To explore the possibility of anticancer drug targeting to lymph metastasis using activated charcoal nanoparticles as a drug delivery carrier, the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin (PYM ) absorbed in activated charcoal nano-particles ( ACH-NP) and the lymph targeting effect of PYM-ACH-NP.Methods PYM-ACH-NP was prepared by mixed the ACH-NP + PYM + saline and shaken for 20 min. The absorbency of PYM on ACH-NP was evaluated. Cervical lymph node metastasis mice model was established by buccal submucosa implantation of a high lymph metastasis cell line U14 cancer cells (5 × 10 /L). PYM was radiolabeled with 125I by the modified chloramine T method. Thirty Kunming mice models burdened with cervical lymph metastasis were randomly divided into control group, PYM-treated group and PYM-ACH-NP-treated group. The animal in each group was injected with 0. 2 mL saline or corresponding drugs (equal to 10 mg/kg PYM) respectively. The radioactivity of PYM in blood, heart, liver, spleen, lung,kidney and cervical lymph node was detected 72 h after administration. Specific radioactivity of each sample was calculated. Results The average diameter of PYM-ACH-NP was 176 nm. The absorbency of PYM on ACH-NP was increased with the increased ratio of ACH-NP to PYM. In PYM-CH-NP group, specific radioactivity of PYM was significantly higher in cervical lymph node ( 148. 72 ± 29. 35 ) cpm/mg than in PYM group ( 10. 17 ±2. 11) cpm/mg ( P <0. 01 ) , meanwhile the specific radioactivity of drug in the blood, heart, liver, spleen, lung and kidney of PYM-CH-NP group was (2. 18 ±0. 39), (1. 19 ±0.21) ,(2.41 ±0.50) , (1.09 ±0.24) , (1.95 ±0.47) and (2.21 ±0.44) cpm/mg respectively, which was significantly lower than in PYM group (17. 22 ± 3. 04) , (2. 48 ± 0. 47 ), (6. 94 ± 1. 38 ), (4. 12 ±0.79), (8. 25 ±2.04), (18. 83 ±3. 89) cpm/mg. The uptake of PYM in the blood, heart

  14. 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)



    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