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PubMed1975

The acute effect of sodium cellulose phosphate on intestinal absorption and urinary excretion of calcium in man.

Intestinal 47Ca absorption has been determined from blood and stool radioactivity after oral administration of the isotope in nine patients before and during cellulose phosphate treatment. Oral administration of 5 g cellulose phosphate concomitant with 47Ca in 100 mg "carrier" calcium as CaCl2 decreased 47Ca absorption by 80 per cent. Cellulose phosphate, 5 g three times daily, decreased urinary excretion of non-radioactive calcium by 47 percent. Urinary magnesium excretion decreased by 47 percent whereas urinary phosphorus excretion increased by 67%. Calcium and magnesium excretion in urine decrease because cellulose phosphate binds divalent cations within the GI tract. The increased phosphorus excretion is probably due to partial hydrolysis of the substance in the gut.

Calcium↗

PubMed1985

Active-site-directed inhibition of 3-hydroxy-3-methylglutaryl coenzyme A synthase by 3-chloropropionyl coenzyme A.

3-Chloropropionyl coenzyme A (3-chloropropionyl-CoA) irreversibly inhibits avian liver 3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase). Enzyme inactivation follows pseudo-first-order kinetics and is retarded in the presence of substrates, suggesting that covalent labeling occurs at the active site. A typical rate saturation effect is observed when inactivation kinetics are measured as a function of 3-chloropropionyl-CoA concentration. These data indicate a Ki = 15 microM for the inhibitor and a limiting kinact = 0.31 min-1. [1-14C]-3-Chloropropionyl-CoA binds covalently to enzyme with a stoichiometry (0.7 per site) similar to that measured for acetylation of enzyme by acetyl-CoA. While the acetylated enzyme formed upon incubation of HMG-CoA synthase with acetyl-CoA is labile to performic acid oxidation, the adduct formed upon 3-chloropropionyl-CoA inactivation is stable to such treatment. Therefore, such an adduct cannot solely involve a thio ester linkage. Exhaustive Pronase digestion of [14C]-3-chloropropionyl-CoA-labeled enzyme produces a radioactive compound which cochromatographs with authentic carboxyethylcysteine using reverse-phase/ion-pairing high-pressure liquid chromatography and both silica and cellulose thin-layer chromatography systems. This suggests that enzyme inactivation is due to alkylation of an active-site cysteine residue.

Acyl Coenzyme A↗

PubMed1991

Improvement in the diagnostic potential of 32P-postlabeling analysis demonstrated by the selective formation and comparative analysis of nitrated-PAH-derived adducts arising from diesel particle extracts.

Two versions of the 32P-postlabeling assay (nuclease P1 and butanol extraction) enhance the detection limit of polycyclic aromatic hydrocarbon (PAH)-modified DNA. Previously published studies suggest that DNA adducts derived from N-substituted aryl compounds are poorly recovered in the nuclease P1 version. In this study, both versions were employed to ascertain whether the apparent differences in sensitivity could be used to select diagnostically for nitroaromatic-DNA adducts derived by treating calf thymus DNA with organic extracts isolated from four diesel and one gasoline vehicle emission particles. We enhanced the formation of nitrated-PAH-derived adducts through xanthine oxidase (XO)-catalyzed nitroreduction of nitrated-PAHs, constituents previously detected in the diesel emissions. Chromatographic mobilities of the XO-derived DNA adducts were compared to adducts detected in calf thymus DNA resulting from rat liver S9-mediated metabolism. All four diesel organic extracts treated with XO resulted in the formation of one major DNA adduct, chromatographically distinct from the multiple DNA adducts detected in the rat liver S9-treated incubations. This adduct was detectable with the butanol extraction but not the nuclease P1 version of the 32P-postlabeling assay and was chromatographically similar to DNA adducts formed following XO nitroreduction of 1-nitropyrene or ascorbic acid treatment of 1-nitro-8-nitroso-pyrene and 1-nitro-6-nitrosopyrene. In contrast, when S9 activation was used, multiple DNA adducts were detected along a diagonal zone of radioactivity and were radioactively labeled with equivalent efficiency irrespective of the assay version employed. The in vitro calf thymus DNA model described in this study enhances the diagnostic potential of the 32P-postlabeling assay through the selective formation of nuclease P1-sensitive N-substituted aryl-derived DNA adducts.

Air Pollutants↗

PubMed1973

Radiation and pregnancy.

Irradiation during pregnancy may occur either as the result of radioactive pollution of the environment, or during a medical procedure using x-rays or radionuclides. While the former is usually unforeseeable, the latter is known and accepted by both physician and patient.Recent statistics estimate that about one quarter of pregnant women have had a radiographic experience during the pregnancy, either for obstetrical reasons or in the course of medical and dental examinations. The amount of radiation delivered to the fetus is in the range of one rad or less. Radionuclidic procedures may result in fetal radiocontamination, chiefly after placental transfer and fetal uptake. Radioiodine, radioactive calcium and selenomethionine are dangerous for the fetus, since they cross the placenta freely and are taken up by fetal tissues. The labelled proteins, radiocolloids and some mercury compounds remain in the maternal compartment and therefore can affect the fetus only through their gamma radiation at some distance from the fetus.The teratogenic effect, the leukemogenic threshold and the lowered resistance to neonatal infections have been demonstrated after irradiation with doses far higher than those encountered during diagnostic applications of ionizing radiation. Statistical data suggest an increase of susceptibility to leukemia in infancy after intra-uterine irradiation at a diagnostic level. Cytogenic analysis may.... offer valuable data for the establishment of the extent of radiation damage.

Chromosome Aberrations↗

PubMed1969

Effect of endotoxin and cortisone on synthesis of ribonucleic acid and protein in livers of mice.

The effect of cortisone and endotoxin, singly and in combination, on ribonucleic acid (RNA) synthesis in livers of adrenalectomized mice was determined. This was accomplished by measuring the incorporation either of inorganic (32)P or of (14)C-orotic acid into the RNA. Under similar conditions, the effect of these agents on the rate of protein synthesis was examined with the use of (14)C-leucine. Bacterial endotoxin was found to augment the uptake of isotope in the RNA and in the protein of the liver. These reactions did not appear to be mediated via the pancreatic hormone insulin, which was found to depress the incorporation of the radioactive compounds into RNA. Cortisone increased the uptake of isotope in liver RNA but depressed the incorporation of leucine into hepatic protein. These results indicate that the previously observed ability of endotoxin to prevent the hormone induction of hepatic enzymes, such as tryptophan oxygenase, is not associated with impaired synthesis of liver RNA or protein.

Adrenalectomy↗

PubMed1984

Radioactivity in blood and urine following intraperitoneal instillation of chromic phosphate in patients with and without ascites.

Systemic distribution of radioactive colloidal chromic phosphate P 32 after intraperitoneal instillation was studied in 10 patients with ovarian or endometrial malignancies. Seven patients without ascites received chromic phosphate P 32 for positive peritoneal washings, rupture of the capsule of the cyst during operation, or minimal Stage III disease. Three patients received chromic phosphate P 32 for recurrent ascites after multiple abdominal paracenteses. Blood and urine radioactivity measurements were performed at selected intervals. There was a clear statistically significant difference (p less than 0.01) between chromic phosphate P 32 activity levels in whole blood, red blood cells, and plasma in patients with and without ascites.

Adult↗

PubMed1977

Metal isotopes used as radioactive indicators of ocular melanoma.

We studied radioactive metal compounds to determine their localization in malignant Greene melanoma in the eye of the Syrian golden hamsters. Scintigraphy of an ocular melanoma was achieved with several radioactive metals: radioactive indium(111In)-bleomycin, radioactive gallium (67Ga)-citrate, and radioactive lead (203Pb)-tris. The 111In-bleomycin had the highest tumor uptake (5.45% dose/g), but not the highest tumor-to-background ratios. The 67Ga-citrate had a maximum tumor uptake of 4.87% dose/g at 48 hours. The 111In-chloride had a melanoma uptake of 2.26% dose/g, while with 203Pbtris the uptake was 1.61% dose/g. These uptake ratios compare favorably with that of radioactive phosphorus (32P)-2.21% dose/g. Noninvasive localization of malignant melanoma in the eye was accomplished with metal isotopes. Detailed analysis of in vitro uptake date by tumor and background tissues revealed the optimum time for scanning and precluded unnecessary trials.

Animals↗

PubMed1986

[Chemical carcinogens and DNA, covalent binding and chemical modifications].

Chemical carcinogenic agents are usually metabolized into final metabolites susceptible to be DNA bound to form adducts. They may also modify DNA bases. Binding may be studied in vitro in the presence of an enzyme system causing the formation of final metabolites or in vivo. Because of the small amount of adducts formed in vivo, they can only be visualized with radioactive compounds or with a 32P marking technique after DNA hydrolysis. A limit of detection of adduct for 10(8) nucleotides has thus been reached for polycyclic compounds. The covalent binding index enables to quantify this binding. The consequences of DNA alterations vary according to the substitutes. In the case of acetyl-aminofluorene, they lead to mutations by shifting of the reading frame (frameshift). Alkylations lead to inaccurate pairing causing pin point mutations or depurinations.

Carcinogens↗

PubMed2001

Differential elution of sodium or potassium dihydrogen- and hydrogenphosphate ions from a sephadex G-15 column with sodium or potassium chloride solution.

When a mixed solution of sodium or potassium dihydrogenphosphate and disodium or dipotassium hydrogenphosphate was eluted from a Sephadex G-15 column with either a sodium or potassium chloride solution, the elution profiles of ions showed that the hydrogenphosphate ion was eluted more rapidly than the dihydrogenphosphate ion. When the sample solutions containing potassium dihydrogenphosphate and/or dipotassium hydrogenphosphate, all of which were supplemented with phosphorus-32-labelled potassium dihydrogenphosphate, were eluted with sodium chloride solution, the elution profiles of radioactivity showed that the dihydrogenphosphate ion changed to hydrogenphosphate ion and vice versa, depending on the pH values of the sample solution and the availability of the cation of the eluent during elution for the phosphate ion to pair with.

Chromatography, Ion Exchange↗

PubMed1980

Absence of phosphates from the myosin heavy chains of striated muscles.

This work aimed to determine whether the heavy chains of myosin from different striated muscle were phosphorylated. Myosin and its heavy chains were prepared from cardiac and skeletal muscles of rats injected in vivo with radioactive phosphates. The results for radioactive phosphate localization indicate the absence of phosphate from pure heavy chains and from any of their purified fragments, whatever the striated muscle used. In addition, phosphates are present in the myosin phosphorylated light chain and in a contaminating protein closely associated to the myosin heavy chain.

Animals↗

PubMed2003

Dose model for stent-based delivery of a radioactive compound for the treatment of restenosis in coronary arteries.

Radiolabeled drug-eluting stents have been proposed recently as a novel method to potentially reduce restenosis in coronary arteries. A P-32 labeled oligonucleotide (ODN) loaded on a polymer coated stent is slowly released in the arterial wall to deliver a therapeutic dose to the target tissue. However, the relatively low proportion of drugs transferred to the arterial wall (<2%-5% typically) raises questions about the degree to which radiolabeled drugs eluted from the stent can contribute to the total radiation dose delivered to tissues. A three-dimensional diffusion-convection transport model is used to model the transport of a hydrophilic drug released from the surface of a stent to the arterial media. Large drug concentration gradients are observed near the stent struts giving rise to a nonuniform radiation activity distribution for the drug in the tissues as a function of time. A voxel-based kernel convolution method is used to calculate the radiation dose rate resulting from this activity build-up in the arterial wall based on the medical internal radiation dose formalism. Measured residence time for the P-32 ODN in the arterial wall and at the stent surface obtained from animal studies are used to normalize the results in terms of absolute dose to tissue. The results indicate that radiation due to drug eluted from the stent contributes only a small fraction of the total radiation delivered to the arterial wall, the main contribution coming from the activity that remains embedded in the stent coating. For hydrophilic compounds with rapid transit times in arterial tissue and minimal binding interactions, the activity build-up in the arterial wall contributes only a small fraction to the total dose delivered by the P-32 ODN stent. For these compounds, it is concluded that radiolabeled drug-eluting stent will not likely improve the performance of radioactive stents for the treatment of restenosis. Also, variability in the delivery efficacy of drug delivery devices makes accurate dosimetry difficult and the drug washout in the systemic circulatory system may yield an unnecessary activity build-up and dose to healthy organs.

Arteries↗

PubMed1988

Rabbit synoviocyte inositol phospholipid metabolism is stimulated by hydroxyapatite crystals.

Inhibition of prostaglandin E2 synthesis partially ameliorates some aspects of synovitis, but joint destruction still progresses. Other aspects of phospholipid metabolism may play a role in synovial tissue pathophysiology. Products of phosphatidylinositol metabolism can activate intracellular processes in response to extracellular stimuli. We asked whether this pathway is activated in synoviocytes in monolayer tissue culture by the addition of hydroxyapatite (HA) crystals in medium. These crystals are found in pathological human synovial fluid. These crystals are associated with the secretion of degradative enzymes and with a destructive arthritis in humans. Rabbit synoviocyte cultures, previously incubated with [3H]inositol to label inositol phospholipids, were stimulated with the addition of hydroxyapatite (180 micrograms/ml) to the cultures. There was enhanced intracellular accumulation of [3H]inositol monophosphate (30-100%) after 4 h. This indicated an increased phospholipase C activity. The radioactivity in [3H]inositol bis- and trisphosphates was too low to reliably measure. The use of [32P]Pi allowed detection of these compounds. In the presence of HA, incorporation of [32P]Pi into phosphatidylinositol, phosphatidylinositol monophosphate, and phosphatidylinositol bisphosphate was increased. In addition, cultures exposed to [32P]Pi during stimulation with HA had an increased content of [32P]inositol monophosphate, bisphosphate, and trisphosphate.

Animals↗

PubMed1984

Evaluation of neutrophil labeling techniques using the chemotaxis radioassay.

Neutrophils isolated from human blood were labeled by various methods and exposed to a chemotactic gradient. The chemotactically functional cells that migrated into the gradient were isolated. The portion of radioactivity of the original cell suspension carried with the chemotactically responsive cells was related to the relative number of migrating cells as determined microscopically. Of the radionuclides used, P-32 diisopropylfluorophosphate (DFP), In-111 oxine, and Tc-99m sulfur colloid provided cell preparations with the highest relative portion of radioactivity confined to functionally intact (chemotactic) neutrophils. Results with Na2(51)CrO4 and with SnCl2-reduced 99mTcO4- were less than optimal. Neutrophils exposed to Ga-67 citrate apparently took up the label and retained chemotactic responsiveness. However, little or no radioactivity was detected in the neutrophils that migrated from the suspensions of Ga-67-labeled cells. The results indicate that the chemotaxis radioassay can yield unique information pertaining to the extent to which a radiotracer is specifically associated with viable neutrophils in a suspension of labeled cells.

Chemotaxis, Leukocyte↗

PubMed1988

[Chemical transformation of radioactive 4-(N-2-chloroethyl-N-methylamino)benzyl-5'-[32P]phosphamides of oligodeoxyribonucleotides during in vivo experiments].

The practical use of reactive oligonucleotide derivatives for complementarily addressed modification of nucleic acids in vivo includes several steps, at which side chemical reactions resulting in a decrease of the modification efficiency may take place. Chemical reactions of 4-(N-2-chloroethyl-N-methylamino)benzyl-5'-[32P]phosphamides of oligodeoxyribonucleotides were studied in vivo. The intermolecular self-alkylation at the reactive residue of the alkylating derivative was found in the precipitate of its lithium salt under acetone at-20 degrees C. The effects of pH, buffer solutions, salts, temperature, phenol, cell culture suspensions, tissue homogenates, etc., on the stability of the derivatives were studied. A sufficient cleavage of the phosphamide bond was observed at pH less than 3. In fresh liver homogenates the nucleolytic degradation of the oligonucleotide part of the reagent was shown to occur. After intraperitoneal injection of mice with radioactive alkylating derivatives up to 50% of the reagent was included into the blood biopolymers within one hour. The covalently linked to the biopolymers oligonucleotide appeared to be highly degraded thereby.

Alkylating Agents↗

PubMed1975

Nonenzymatic acetylation of histones with acetyl phosphate and acetyl adenylate.

Nonenzymatic acetylation of calf-thymus lysine- and arginine-rich histones was demonstrated to occur when these proteins were incubated with [14C]acetyl phosphate and [14C]acetyl adenylate. The levels of acetylation depend on both pH and on reagent concentration. When acetyl [33P]phosphate and acetyl [3H]adenylate were used as reagents, we found neither histone phosphorylation nor adenylylation. Most of the radioactivity of 14C-labeled acetylated histones was recovered as Ne-acetyllysine. Furthermore, only a small amount of O-bound radioactivity was released by the 14C-labeled acetylated arginine-rich histone during treatment with hydroxylamine. Experiments on the acetylation of histones, in the presence of increasing salt concentration, gave different results for the two acetylating agents.

Acetates↗

PubMed1993

Immunoaffinity concentration of human lung DNA adducts using an anti-benzo[a]pyrene-diol-epoxide-DNA antibody. Analysis by 32P-postlabelling or ELISA.

DNA, isolated from 15 human lung autopsy samples, was examined for the presence of polycyclic aromatic hydrocarbon (PAH) DNA adducts. Using the nuclease P1 modification of the 32P-postlabelling technique, between 1 and 12 adducts/10(8) nucleotides were detected prior to immunoconcentration. Autoradiograms from most of the samples revealed a diagonal smear of radioactivity consistent with complex mixture (cigarette smoking) DNA damage. The DNA samples were digested to oligonucleotides, made single-stranded and subsequently applied to immunoaffinity columns containing immobilised anti-benzo[a]pyrene (B(a)P)-7,8-diol-9,10-epoxide (BPDE) DNA polyclonal rabbit antibody. The material remaining bound to the column, in addition to that passing through, was analysed using both ELISA and 32P-postlabelling techniques. Column-bound adducts comprised between 0% and 78% of any particular sample. Immunoconcentration, followed by 32P-postlabelling of the material which had been bound to the column, revealed the presence of a number of discrete adduct spots in autoradiograms of the more heavily adducted samples. Sample DNA not retained by the columns was also analysed; the chromatographic pattern obtained was a dense zone of radioactive material migrating from the origin. This evidence suggests that the composition of PAH-DNA adducts found in human lung samples exhibits wide inter-individual variation.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

PubMed2005

Partitioning of radionuclides and trace elements in phosphogypsum and its source materials based on sequential extraction methods.

Phosphogypsum is a waste produced by the phosphate fertilizer industry. Although phosphogypsum is mainly calcium sulphate dihydrate, it contains elevated levels of impurities, which originate from the source phosphate rock used in the phosphoric acid production. Among these impurities, radionuclides from 238U and 232Th decay series are of most concern due to their radiotoxicity. Other elements, such as rare earth elements (REE) and Ba are also enriched in the phosphogypsum. The bioavailability of radionuclides (226Ra, 210Pb and 232Th), rare earth elements and Ba to the surrounding aquatic system was evaluated by the application of sequential leaching of the phosphogypsum samples from the Brazilian phosphoric acid producers. The sequential extraction results show that most of the radium and lead are located in the "iron oxide" (non-CaSO4) fraction, and that only 13-18% of these radionuclides are distributed in the most labile fraction. Th, REE and Ba were found predominantly in the residual phase, which corresponds to a small fraction of the phosphate rock or monazite that did not react and to insoluble compounds such as sulphates, phosphates and silicates. It can be concluded that although all these elements are enriched in the phosphogypsum samples they are not associated with CaSO4 itself and therefore do not represent a threat to the surrounding aquatic environment.

Barium↗

PubMed1994

Synoviorthesis with colloidal 32P chromic phosphate for the treatment of hemophilic arthropathy.

Between 1977 and 1992, we performed ninety-two synoviortheses (destruction of synovial tissue by intra-articular injection of a radioactive agent) on forty-eight patients who had a severe congenital disorder of hemostasis and chronic hemophilic synovitis that was resistant to conventional treatment. Colloidal 32P chromic phosphate was injected intra-articularly: 1.0 millicurie for knees and 0.5 millicurie for other joints. The duration of follow-up ranged from one to fifteen years. The frequency and importance of bleeding decreased in most of the patients. The range of motion of half of the joints remained stable or improved and that of the other half continued to decrease. Radiographic scores worsened progressively despite the decreased frequency of hemarthrosis. In most patients, the extra-articular leakage of the radioactive agent was slight. Chromosome breakages were observed almost exclusively in patients who were seropositive for human immunodeficiency virus and in whom the CD4-lymphocyte count was decreased from normal. The patients' level of satisfaction with the results was high.

Adolescent↗