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Biomedical subjects

A T Vaughan

Publications and source records attributed to A T Vaughan.

At least 55 records · Page 3Linked to original sources
PubMed1989

Yttrium-90-EDTMP: a radiotherapeutic agent in the treatment of leukaemias.

Yttrium-90 chelated by the tetraphosphonate EDTMP achieved a high uptake in bone and a rapid clearance from all soft tissues compared with 90Y nitrilotriacetate, citrate and acetate. The biological half-life of 90Y in the bone was greater than 72 h, but the quantity, and therefore dose, could be reduced by 50% using repeated, non-toxic chelation therapy with the calcium salt of DTPA. This treatment should be able to supplement current treatments for leukaemia where the dose of external beam radiation is associated with considerable morbidity.

Animals↗
PubMed1989

Conformational changes in chromatin structure induced by the radioprotective aminothiol, WR 1065.

WR 1065, 2-[(minopropyl) amino] ethanethiol is an effective scavenger of free radicals. When present during irradiation it reduces cellular DNA damage as analysed by alkaline elution from filters. The same technique indicates that without irradiation, WR 1065 has no effect of DNA integrity. Using nucleoid analysis, where DNA damage is detected at the level of replicon clusters, WR 1065 distorts replicon supercoiling without breaking the DNA molecule. This confirmational change in nucleoid structure occurs with no detectable change in nucleoid protein content. It is proposed that perturbation of replicon supercoiling affects the process of normal DNA synthesis and strand break rejoining, allowing a longer time for the accurate repair of DNA damage.

Animals↗
PubMed1989

The use of flow cytometry in the detection of radiation- and chemically induced nuclear damage in human mononuclear cells.

Viable human mononuclear cells were treated with three known genotoxic agents, gamma radiation, bleomycin sulphate (BLM) and acetylaminofluorene (AAF). Following treatment, a high salt concentration buffer was used to disrupt the cells, releasing supercoiled DNA structures known as nucleoids. The nucleoids were treated with the fluorescent dye ethidium bromide, and exposed to a beam of laser light within a fluorescence activated cell sorter (FACS). Treatment of human mononuclear cells with the genotoxic agent BLM or with radiation prior to cell disruption and staining produced a dose-dependent increase in light scatter. However, treatment of human mononuclear cells with AAF, which requires metabolic activation, produced a dose response in light scatter that proved reversible and was not dependent on the formation of reactive metabolites. In the case of BLM and radiation, these changes in light scatter are interpreted as structural alterations in the nucleus partially induced by free radical attack, and thus provide an indication of the DNA damaging ability of the agent used. In contrast the reversible changes in light scatter seen with AAF may only indicate the ability of this agent to penetrate the genome. The significance of this reversible interaction with respect to genotoxicity is not known.

2-Acetylaminofluorene↗
PubMed1988

Antibodies labeled with 199Au: potential of 199Au for radioimmunotherapy.

We have investigated the direct labeling of antibodies with gold isotopes as an alternative to iodine and chelated metals for antibody guided therapy. Both 199Au and 195Au complex anions in citrate-buffered solutions (pH 3.8-7.2) are rapidly bound to sheep polyclonal anti-CEA with high efficiency. After purification by gel filtration, the antibody retained 80% of the bound gold for upto 6 days at 4 degrees C and was reactive with its target antigen in a solid phase assay. Incubation with the gold binding substance D-penicillamine reduced labeling efficiency and stability.

Animals↗
PubMed1988

Structural damage to lymphocyte nuclei by H2O2 or gamma irradiation is dependent on the mechanism of OH. radical production.

Normal human lymphocytes were exposed to OH. radicals produced indirectly by exposure to H2O2 or directly by gamma irradiation. Using a flow cytometry technique to measure changes in nucleoid size, it was found that generation of OH. in each system produced a characteristic relaxation in nuclear supercoiling. Exposure of cells to H2O2 produced a metal-dependent step-wise relaxation in extracted nucleoids, while gamma irradiation induced a gradual dose-dependent increase in nucleoid size. The site-specific metal-dependent changes produced in lymphocytes incubated in H2O2 should also occur in gamma irradiated cells, but the characteristic effects on nuclear supercoiling would not be detected within the background of random DNA damage. The importance of metals in maintaining the supercoiled loop configuration of DNA within the protein matrix suggests that free radical damage at metal locations may be particularly toxic for the cell.

Adult↗
PubMed1987

Limitations to the killing of tumours using radiolabelled antibodies.

We have calculated the minimum requirements for effective therapy using intravenously administered, tumour-directed antibodies labelled with either iodine 131 or yttrium 90. A lethally large amount of either radionuclide would be required to achieve tumour destruction. At least a 10-fold increase in tumour uptake is necessary to combine tumour destruction with a survivable whole-body dose. The required improvement in specific uptake can be substantially reduced by accelerating the excretion of radioactivity outside the tumour. For all situations studied, yttrium 90 is superior to iodine 131 as a cytotoxic label.

Antibodies, Monoclonal↗
PubMed1987

Antibodies labeled with metallic radionuclides: influence of nuclide chemistry on dose distribution.

An antibody with human CEA specificity has been labeled with either yttrium-90, scandium-47, or indium-111, via a diethylenetriamine pentaacetic acid (DTPA) link covalently bound to the protein. The clearance of these proteins from the blood of mice can be described by a single exponential; the half-life decreases in the order indium-111 greater than yttrium-90 greater than scandium-47. Associated with the blood clearance is an uptake of radioactivity into the liver; scandium-47 has the highest concentration, indium-111 has the least, and yttrium-90 is intermediate. There is no correlation between these results and the equilibrium stability constants of the metals with DTPA-like ligands. The results obtained show that, in vivo, scandium-47 and yttrium-90 are more easily displaced from DTPA by other ions than is indium-111. They also show that free DTPA is able to extract yttrium-90 and scandium-47, but not indium-111, from the liver of treated animals, indicating that indium-111 is resistant to ligand exchange reactions in vivo. These data indicate that 1) the equilibrium stability constant is not a good indicator of the in vivo stability of metal-labeled proteins and 2) it is possible to manipulate the ion distribution and therefore the dose from scandium-47 and yttrium-90 after injection of the labeled proteins.

Animals↗
PubMed1987

Measurement of DNA damage in mammalian cells using flow cytometry.

A technique for the detection of DNA damage induced by radiation insult has been developed. Cells were lysed with a buffer containing 2 M sodium chloride to release the DNA in a supercoiled form, the nucleoid. These were stained with the DNA intercalating dye, ethidium bromide, and exposed to laser light within a flow cytometer. Scattered and fluorescent light was analyzed from the laser/nucleoid interaction following irradiation of viable cells with gamma rays. The addition of ethidium bromide to prepared nucleoids caused a reduction in scattered light due to condensation of the nucleoid. Irradiation of cells prior to nucleoid production and ethidium bromide treatment restricted this condensation and produced a dose-dependent increase in laser scatter. Nucleoids derived from human lymphocytes showed enhanced light scatter from 5 Gy, compared to Chinese hamster ovary (CHO) fibroblasts where doses above 10 Gy were required. Up to 30 Gy CHO nucleoids showed a dose-dependent reduction in the ethidium bromide fluorescence. This technique allows detection of altered light scattering and fluorescent behavior of nucleoids after cellular irradiation; these may be related to structural changes within the nucleus induced by the radiation. The use of flow cytometry compared to other methods allows a rapid analysis of nuclear damage within individual cells.

Animals↗
PubMed1986

Limitations in localising and killing tumours using radiolabelled antibodies.

Radiolabelled antibodies against tumours are being studied intensely for their use in tumour detection and tumour destruction. Although a host of different tumour types can be localised, results at present are no better than other scanning techniques. In addition no tumour has been eradicated using labelled antibodies. Dramatic improvements will depend upon a fuller understanding of tumour cell biology and optimising all the parameters involved.

Antibodies, Neoplasm↗
PubMed1985

The production and biological distribution of yttrium-90 labelled antibodies.

An antibody has been labelled with generator produced yttrium-90 (90Y) via a chelating group (DTPA) covalently attached to the protein. An average yield for protein labelling of 76% was obtained. The blood clearance of iodinated and 90Y labelled antibody is similar in rats and mice. However, in contrast to iodine, the yttrium label accumulates in the liver after release from the protein.

Animals↗
PubMed1983

Improved tumour localisation using indium-111 labelled antibodies.

Immunoglobulin G (IgG) antibodies to carcinoembryonic antigen (CEA) were labelled with radioactive indium (111In) or iodine (131I) and a comparison made of their value in locating CEA producing tumours. Eleven patients given 111In-anti-CEA had 31 tumours as judged by a combination of all techniques. Of these, 28 were detected by 111In-anti-CEA and 26 by conventional clinical techniques. Five of the patients also received 131I-anti-CEA. These patients had 15 tumour areas. Thirteen were detected by 111In and eight by 131I. 111In also produced a better signal to noise ratio in the scans and thereby showed lesions with greater certainty. In addition, the 111In isotope continued to accumulate in the tumour areas for considerably longer than 131I. Absorbed doses (whole body) were similar for both isotopes. The results show that antibody scanning is greatly improved by using 111In as the radiolabel in place of 131I and should allow the detection of smaller or deeper lesions.

Carcinoembryonic Antigen↗
PubMed1983

Synthesis of cathepsin B by cells derived from the HL60 promyelocytic leukaemia cell line.

Cells of the human promyelocytic HL60 line were induced to differentiate into granulocyte-like cells with dimethylsulphoxide (DMSO) or macrophage-like cells with 12-0-tetradecanoylphorbol-13-acetate (TPA). The synthesis of Cathepsin B by these cells was studied by immunoperoxidase staining and assay of cell lysates using the fluorimetric substrate benzoyloxycarbonyl-phenylanalyl-arginine-4-methyl-7-coumarylamide. Only 2-5% of the uninduced HL60 cells and DMSO-induced cells were immunohistochemically positive for Cathepsin B, compared with over 80% of the TPA-induced cells. Cathepsin B activity was lowest in the lysates of uninduced HL60s. DMSO-induced cells contained 1.5-2-fold the enzyme activity of HL60s and TPA-induced cell lysates demonstrated 5-14-fold the activity of uninduced HL60s. Induction of Cathepsin B synthesis was therefore associated with differentiation of the promyelocytes into cells of the monocyte/macrophage type, but not granulocyte-like cells. Cathepsin B was located immunohistochemically in human palatine tonsils. The enzyme was only demonstrated within macrophages in these tissues. Cathepsin B may therefore be a useful immunohistochemical marker for malignant and nonmalignant cells of the monocyte/macrophage lineage.

Cathepsin B↗
PubMed1982

The specific inhibition of cellular clonogenic proliferation using 211At labelled lectins and antibodies--I.

A monoclonal antibody with a broad anti-human leucocyte specificity, designated BK 19.45 and the plant lectin Concanavalin A have been labelled with the alpha-emitting cyclotron produced radiohalogen astatine-211. Both human and murine tumour cell lines and human leukemic bone marrow samples have been specifically labelled with these radioactive proteins. In all cases the amount of 211At bound to the cells is directly correlated with a decrease in cellular reproductive potential as shown by the ability of the cells to proliferate in a clonogenic assay. For the labelled monoclonal antibody experiments, the radiation dose needed to yield 37% cell survival, the D37 dose, may be achieved with an average of 12 211At atoms/cell.

Animals↗
PubMed1982

The in vivo fate of a 211At labelled monoclonal antibody with known specificity in a murine system.

A monoclonal antibody reactive against the human transferrin receptor has been labelled with the alpha and X ray emitting isotope Astatine 211. The labelling procedure does not affect the ability of the product to bind to the transferrin receptor on the human leukemic cell line HL60. Using a direct binding assay, 211At labelled antibody can be specifically inhibited from binding to its target cells by excess unlabelled antibody. Furthermore, the binding inhibition demonstrated in this system correlates to enhanced clonogenic survival of these cells, indicating that very few atoms of 211At/cell are required for cell death. Data obtained from labelled antibody injected into mice show that the labelled product in serum retains the ability to bind to HL60 cells in vitro, although tissue distributions of the injected activity implies that some of the radiolabel is lost from the protein. Despite this loss of label, preliminary experiments on the localization of labelled antibody to HL60 cells growing s/c in nude mice show that tumor tissue has a higher specific activity than all other tissues, other than blood, after 12 hours. This suggests that further work on the nature of label degradation in vivo is warranted in the context of potential therapeutic and diagnostic studies.

Animals↗
PubMed1981

Mice treated with strontium 90: an animal model deficient in NK cells.

Treatment of BALB/c mice with radioactive isotopes of the bone-seeking element strontium reduces the percentage of specific NK-cell cytotoxicity to only 2.6%, compared with 13.6% for normal BALB/c and 36.3% for athymic (nude) BALB/c. The syngeneic plasmacytoma NS-1 was used as target in a 4th in vitro NK-cell microassay. Marrow cellularity in treated mice is reduced to 12.5% of controls, but haemopoietic and stem-cell functions are taken over by the spleen and the peripheral blood picture remains relatively normal. Allogenic (H-2k) tumour transplants are rejected normally with good anti-H-2k alloantibody response. Haemopoietic and T- and B-cell functions are therefore substantially intact, and the defect seems confined to NK cells. In vivo, after s.c. inoculation of 10(6) NS-1 cells, 8/12 controls grew a solid tumour after a mean delay of 30.5 +/- 1.25 (s.e.) days, whereas 5/6 90Sr-treated mice grew the tumours after a delay of only 10.5 +/- 1.8 days. This markedly reduced delay in the 90Sr-treated mice lends support to suggestions that NK cells play an important role in resisting the establishment of tumour foci (i.e. in antitumour surveillance). Mice treated with 90Sr could be useful in evaluating the in vitro role of NK cells.

Animals↗