RESUMO
PURPOSE: Neoplastic meningitis is often the final outcome of disseminated cancer and is rapidly lethal. Its limited treatment relies on systemic or intrathecal chemotherapy with methotrexate (MTX) or thiotepa. When 5-iodo-2'-deoxyuridine labeled with (125)I ((125)IUdR) is incorporated into the DNA of mitotic tumor cells, the Auger electrons emitted during iodine decay are highly cytotoxic. The radiotherapeutic efficacy of (125)IUdR administered intrathecally has also been established in animals bearing spinal cord tumors, and MTX is known to potentiate the response. This approach has not been tested in the clinic. METHODS: A 44-year-old woman, with locally advanced pancreatic cancer, was treated for three years with complete systemic remission, but then relapsed with cytologically proven neoplastic meningitis. The patient was given four successive intrathecal injections of MTX (10 mg) every 12 h and, with the fourth dose, 1850 MBq (125)IUdR, followed by four additional MTX doses. The response was monitored by cytology and CA19.9 (carbohydrate antigen 19.9) levels in the cerebrospinal fluid (CSF) as well as by clinical status of the patient. RESULTS: The follow-up of cytology and CA19.9 levels in the CSF showed dramatic improvement within 26 days followed by a biological relapse on Day +36. There was no evidence of local central nervous system toxicity. Three months later, neoplastic meningitis recurred and meningeal tumor infiltration was observed on magnetic resonance imaging. Six months after MTX-(125)IUdR treatment, the patient died. CONCLUSION: (125)IUdR treatment proved to be feasible without acute neurological toxicity and seemed to have produced a biological response. This attempt provides the basis for designing prospective clinical trials.
Assuntos
Antibióticos Antineoplásicos/uso terapêutico , Idoxuridina/uso terapêutico , Neoplasias Meníngeas/tratamento farmacológico , Neoplasias Meníngeas/radioterapia , Metotrexato/uso terapêutico , Compostos Radiofarmacêuticos/uso terapêutico , Adulto , Antibióticos Antineoplásicos/administração & dosagem , Antígenos Glicosídicos Associados a Tumores/líquido cefalorraquidiano , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/radioterapia , Neoplasias Encefálicas/secundário , Terapia Combinada , Resistencia a Medicamentos Antineoplásicos , Evolução Fatal , Feminino , Humanos , Idoxuridina/administração & dosagem , Injeções Espinhais , Radioisótopos do Iodo , Neoplasias Meníngeas/secundário , Metotrexato/administração & dosagem , Recidiva Local de Neoplasia , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/patologia , Compostos Radiofarmacêuticos/administração & dosagemRESUMO
A total of 6,500 mosquitoes were identified during a two-year survey (1999-2001) in Lebanon, and these belonged to twelve species: Culex pipiens, Cx. laticinctus, Cx. mimeticus, Cx. hortensis, Cx. judaicus, Aedes aegypti, Ae. cretinus, Ochlerotatus caspius, Oc. geniculatus, Oc. pulchritarsis, Culiseta longiareolata and Anopheles claviger. Culex pipiens was the most predominant species in Lebanon, collected indoors and outdoors. It was continuously abundant and active throughout the year. Culex judaicus was a small and rare mosquito and it is reported to occur for the first time in Lebanon. On the coastal areas, Ochlerotatus caspius was very common, and proved to be a complex of species as two forms were detected. One of the vectors of malaria, Anopheles claviger, was collected from May to September, from eight sites in Lebanon. Its breeding sites were restricted to fresh, cool, and clean water in pools and wells. Most of these breeding sites were in the populated Metn County where a few indigenous cases of malaria were reported from 1997-2000. This shows that the reported malaria cases were not imported, but caused by the bites of locally infected Anopheles females.
Assuntos
Aedes/classificação , Culex/classificação , Ochlerotatus/classificação , Animais , Feminino , Geografia , Líbano , Masculino , Densidade Demográfica , Estações do AnoRESUMO
An iodinated (125I/127I) ethidium derivative (3,8-diamino-5-[6'-(p-iodobenzoylamino)-4'-azahexyl]-6-phenylphenanthridinium chloride hydrochloride) was synthesized and characterized. The labeling yield of the 125I-labeled derivative was 75% for carrier-free 125I, with a radiochemical purity of 95%. The incubation of iodoethidium with calf thymus DNA resulted in a substantial enhancement of fluorescence yield, indicating the intercalation of this compound into DNA. In the presence of iodoethidium, the nuclei of methanol-treated mammalian cells fluoresced, while those of viable cells did not (since the plasma membrane is impermeable to iodoethidium). When viable cells were incubated with the reduced form of the derivative, 125I/127I-dihydroethidium traversed the plasma membrane, was oxidized in the cytoplasm, and intercalated into nuclear DNA. Finally, we tested the hypothesis that larger malignant solid tumors, containing a relatively greater percentage of degenerating permeable cells, can be targeted with 125I-ethidium. In-vivo studies demonstrated a small but positive correlation (R = 0.72) between tumor volume and the uptake of the derivative. Because of the ubiquitous presence of abnormal permeable cells and necrosis in tumors, our results support the belief that radiolabeled DNA-intercalating or DNA-binding molecules may be of diagnostic and therapeutic value for a variety of solid tumors in humans.
Assuntos
Etídio/análogos & derivados , Radioisótopos do Iodo , Compostos Radiofarmacêuticos/síntese química , Células Tumorais Cultivadas/metabolismo , Animais , DNA/metabolismo , Sistemas de Liberação de Medicamentos , Etídio/metabolismo , Camundongos , Oxirredução , Compostos Radiofarmacêuticos/metabolismoRESUMO
PURPOSE: To address the cytotoxic effects of DNA-incorporated (125)I in Chinese hamster V79 lung fibroblasts under various scavenging conditions. METHODS: The toxic effects of DNA-incorporated 5-[(125)I]iodo-2'-deoxyuridine ((125)IdUrd) were assessed by the colony-forming assay with cells incubated in medium containing serum and/or dimethyl sulphoxide (DMSO). Experiments were carried out at 0.3 or -135 degrees C. RESULTS: When (125)I decays were accumulated at 0.3 degrees C in 10% serum 0, 5 or 10% DMSO, no radioprotection was afforded by 5% DMSO, while the dose modification factor (DMF) for 10% DMSO was 2.0. For cells accumulating decays at 135 degrees C in the presence of 5 or 10% serum, DMSO was radioprotective (DMF= 1.8-1.9). D(0) obtained at each serum concentration correlated strongly (R=0.999) with the scavenging capacity of DMSO. Under these experimental conditions, 10% serum is approximately 3.6 times more protective than 5% serum. CONCLUSIONS: The contribution of indirect mechanisms to the toxicity of (125)I decaying within mammalian cell nuclear DNA can be demonstrated not only with DMSO, but also with the hydroxy radical scavengers present in serum.
Assuntos
DNA/toxicidade , Idoxuridina/toxicidade , Radioisótopos do Iodo/toxicidade , Compostos Radiofarmacêuticos/toxicidade , Animais , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Temperatura Baixa , Cricetinae , Cricetulus , Crioprotetores/farmacologia , Meios de Cultura , DNA/metabolismo , Dimetil Sulfóxido/farmacologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibroblastos/efeitos da radiação , Congelamento , Idoxuridina/metabolismo , Compostos Radiofarmacêuticos/metabolismoRESUMO
The decay of iodine-125 (125I) is accompanied by the emission of low-energy electrons that dissipate most of their energy in approximately 10 nm from the decay site. In mammalian cells, the .OH generated by these electrons are also confined to a small volume. Iodine-125 is thus an excellent probe for assessing the radiobiologic effects produced by .OH in close proximity to the site of a decaying atom. We have compared in pUC19 plasmids (naked DNA) and in Chinese hamster V79 lung fibroblasts (chromatin) the modulation by the .OH scavenger dimethyl sulfoxide (DMSO) of 125I-induced DNA double-strand breaks (DSB). The data indicate that DMSO cannot protect plasmid DNA against DSB damage from 125I decaying within a few angstroms from DNA. However, DMSO attenuated DSB production in V79 cells following the decay of DNA-incorporated 125I, thus suggesting that chromatin structure fosters some DSB formation by indirect mechanism(s). DSB production depends on the environment and/or conformation of DNA. Consequently, current biophysical modeling of DNA damage that is based on naked and non-compacted DNA is inadequate for explaining radiobiologic effects at the cellular level.
Assuntos
Cromatina/genética , Cromatina/efeitos da radiação , Dano ao DNA/efeitos da radiação , DNA/química , DNA/efeitos da radiação , Dimetil Sulfóxido/farmacologia , Elétrons/efeitos adversos , Radioisótopos do Iodo/efeitos adversos , Conformação de Ácido Nucleico , Protetores contra Radiação/farmacologia , Animais , Fenômenos Biofísicos , Biofísica , Cricetinae , Fibroblastos/efeitos da radiação , Radioisótopos do Iodo/farmacocinética , Pulmão/citologia , Modelos Teóricos , PlasmídeosRESUMO
We examined whether the administration of methotrexate (MTX) prior to the injection of 5-[125I]iodo-2'-deoxyuridine (125IUdR) in rats with intrathecal (i.t.) TE-671 human rhabdomyosarcoma would enhance 125IUdR uptake by tumor cells and augment the therapeutic efficacy of this Auger-electron-emitting radiopharmaceutical. TE-671 cells were exposed in vitro to medium +/- MTX, and the percentage of cells in various phases of the cell cycle and the uptake of 125IUdR assessed. In addition, nude rats were injected i.t. with TE-671 cells and later infused i.t. with saline or MTX for 24 h prior to 125IUdR injection, and the radioactivity associated with their spinal cords was determined. Exposure of tumor cells in vitro to MTX leads to an increase in the uptake of 125IUdR as a consequence of both a rise in the absolute uptake per cell and an increase in the percentage of S-phase cells. A corresponding increase of radioactivity within the spinal cords of tumor-bearing rats also occurs in the presence of MTX. Tumor-bearing animals were infused/injected with MTX and/or 125IUdR, and the onset of paralysis was determined as a function of time. We find that: (i) MTX infusion leads to a slight increase in time to onset of paralysis (median [M] = 24 vs. 22 days, p = 0.79); (ii) 125IUdR injection results in a statistically significant delay (p < 0.01) in the onset of paralysis (M= 39 days); (iii) MTX administration prior to 122IUdR injection further increases the therapeutic efficacy (M = 45 days).
Assuntos
Elétrons/uso terapêutico , Idoxuridina/uso terapêutico , Radioisótopos do Iodo/uso terapêutico , Meningite/tratamento farmacológico , Meningite/radioterapia , Metotrexato/farmacologia , Inibidores da Síntese de Ácido Nucleico/uso terapêutico , Compostos Radiofarmacêuticos/uso terapêutico , Rabdomiossarcoma/complicações , Animais , Humanos , Idoxuridina/administração & dosagem , Idoxuridina/farmacocinética , Injeções Espinhais , Radioisótopos do Iodo/administração & dosagem , Radioisótopos do Iodo/farmacocinética , Metotrexato/administração & dosagem , Inibidores da Síntese de Ácido Nucleico/administração & dosagem , Inibidores da Síntese de Ácido Nucleico/farmacocinética , Paralisia/induzido quimicamente , Compostos Radiofarmacêuticos/administração & dosagem , Compostos Radiofarmacêuticos/farmacocinética , Ratos , Ratos Nus , Rabdomiossarcoma/veterinária , Fase S/efeitos da radiação , Medula Espinal/química , Distribuição TecidualRESUMO
OBJECTIVES: To define patterns of treatment among contemporary patients undergoing radical prostatectomy, interstitial radiation, and external beam radiation for prostate cancer. METHODS: We analyzed 291 consecutive patients (Stage T1-T3NXM0) who underwent definitive local treatment for prostate cancer with radical prostatectomy, interstitial seed implantation, or external beam radiation. Patients were stratified into three risk groups based on clinical T stage, serum prostate-specific antigen level at diagnosis, and biopsy Gleason score. The frequency of additional treatments, including androgen deprivation and external beam radiation, given within 3 months of initial local therapy was assessed. Patterns of care were compared and adjusted for risk. RESULTS: Of the 291 patients, 107 (36. 8%) underwent radical prostatectomy, 94 (32.3%) underwent interstitial seed implantation, and 90 (30.9%) underwent external beam radiation. Use of combination therapy differed significantly according to the type of initial local treatment and risk category. No patient in the low-risk group received combination therapy. For patients in the intermediate and high-risk groups, the frequency of combination therapy was significantly lower in the radical prostatectomy group when compared with either the interstitial seed implantation (P <0.001 and P <0.02, respectively) or external beam radiation group (P <0.001 and P <0.001, respectively). CONCLUSIONS: There are significant differences in resource utilization for contemporary patients undergoing definitive local therapy for prostate cancer. These differences may have a significant effect on treatment cost and morbidity, and they will likely make short-term comparisons between different treatment modalities difficult because of the high use of androgen deprivation in men treated with radiation therapy.
Assuntos
Braquiterapia/economia , Braquiterapia/estatística & dados numéricos , Reembolso de Seguro de Saúde , Prostatectomia/economia , Prostatectomia/estatística & dados numéricos , Neoplasias da Próstata/radioterapia , Neoplasias da Próstata/cirurgia , Terapia Combinada/economia , Planos de Pagamento por Serviço Prestado , Alocação de Recursos para a Atenção à Saúde , Sistemas Pré-Pagos de Saúde , Hospitais Universitários/economia , Humanos , Masculino , Medicare , Estadiamento de Neoplasias , Neoplasias da Próstata/classificação , Medição de Risco , São FranciscoAssuntos
Tumores Neuroendócrinos/radioterapia , Octreotida/uso terapêutico , Compostos Radiofarmacêuticos/uso terapêutico , Núcleo Celular/metabolismo , Elétrons , Humanos , Radioisótopos de Índio/farmacocinética , Radioisótopos de Índio/uso terapêutico , Tumores Neuroendócrinos/diagnóstico por imagem , Octreotida/farmacocinética , Cintilografia , Compostos Radiofarmacêuticos/farmacocinéticaRESUMO
UNLABELLED: The induction of in vitro morphological transformation in C3H 10T1/2 cells by 99mTc-Cardiolite (contents of Cardiolite kit [hexakis(2-methoxyisobutylisonitrile) and other components] plus (99m)Tc generator eluate) was examined. METHODS: Cells were grown for 48 h in the presence of 99mTc-Cardiolite or decayed 99mTc-Cardiolite (99mTc-Cardiolite after 1 wk of storage), and cell survival and transformation were assessed by the colony-forming and focus assays, respectively. X-ray was used as a reference for radiation effects, and 20-methylcholanthrene was used as a positive control for focus formation. RESULTS: Exposure of cells to 99mTc-Cardiolite results in a transformation frequency that is not significantly different from that induced by the volume equivalent of decayed 99mTc-Cardiolite. The number of foci per viable cell increases linearly from approximately 0.17 x 10(-4) in the untreated control to 1.7 x 10(-4) at 37 kBq/mL and 30 x 10(-4) at 1100 kBq/mL 99mTc-Cardiolite or its decayed 99mTc-Cardiolite volume equivalent. Furthermore, exposure of cells to low extracellular concentrations of 99mTc-Cardiolite or decayed 99mTc-Cardiolite (cell survival, > or =88%) induces an approximately 20-fold greater number of transformants per viable cell than that observed after 0.5 Gy x-irradiation, a dose that causes the same level of toxicity. CONCLUSION: Radioactive and decayed 99mTc-Cardiolite induce morphological transformation of C3H 10T1/2 cells in vitro. The underlying mechanism does not seem to be related to the radiation effects of decaying 99mTc but to chemical(s) present in the 99mTc-Cardiolite kit.
Assuntos
Sobrevivência Celular/efeitos da radiação , Transformação Celular Neoplásica , Compostos Radiofarmacêuticos/toxicidade , Tecnécio Tc 99m Sestamibi/toxicidade , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta à Radiação , Metilcolantreno/toxicidade , Camundongos , Raios XRESUMO
OBJECTIVE: To determine if benign testicular tumours can be identified clinically and ultrasonographically before surgery. PATIENTS AND METHODS: The clinical and ultrasonographic findings of six patients with testicular adenomatoid tumours and 16 with testicular germ cell tumours were assessed retrospectively and compared. RESULTS: All adenomatoid tumours were characterized on physical examination by a well-defined, painful and unfixed nodule, contrasting with the painless and ill-defined malignant lesions. On ultrasonography, of the six adenomatoid tumours, two appeared hypoechoic, one hyperechoic, two isoechoic and one was normal, whereas none of the 16 tumours appeared normal or isoechoic. CONCLUSIONS: While isoechogenicity was not apparent in the group of testicular malignancies, the two groups had hypo- and hyperechoic patterns. Small, superficial, painful and unfixed testicular tumours that appear isoechoic on ultrasonography should be biopsied through an inguinal approach, with frozen sections assessed, instead of the patients undergoing immediate radical orchidectomy.
Assuntos
Tumor Adenomatoide/diagnóstico por imagem , Neoplasias Testiculares/diagnóstico por imagem , Tumor Adenomatoide/cirurgia , Adulto , Humanos , Masculino , Pessoa de Meia-Idade , Orquiectomia/métodos , Estudos Retrospectivos , Neoplasias Testiculares/cirurgia , UltrassonografiaRESUMO
To elucidate the nature and kinetics of DNA strand breaks caused by low-energy Auger electron emitters, we compared the yields of DNA breaks in supercoiled pUC19 DNA in the presence of the (.)OH scavenger dimethyl sulfoxide (DMSO) after the decay of (125)I (1) in proximity to DNA after minor-groove binding ((125)I-iodoHoechst 33342, (125)IH) and (2) at a distance from DNA ((125)I-iodoantipyrine, (125)IAP). DMSO is efficient at protecting supercoiled plasmid DNA from the decay of (125)I free in solution (dose modification factor, DMF = 59 +/- 4) and less effective when the (125)I decays occur close to DNA (DMF = 3.8 +/- 0.3). This difference is due mainly to the inability of DMSO to protect DNA from the double-strand breaks produced by groove-bound (125)I (DMF = 1.0 +/- 0.2). Additionally, the fragmentation of plasmid DNA beyond the production of single-strand and double-strand breaks that is seen after the decay of (125)IH and not (125)IAP (Kassis et al., Radiat. Res. 151, 167-176, 1999) cannot be modified by DMSO. These results demonstrate that the mechanisms underlying double-strand breaks caused by the decay of (125)IH differ in nature from those caused by the decay of (125)IAP.
Assuntos
Dano ao DNA , DNA de Cadeia Simples/efeitos da radiação , Radioisótopos do Iodo , Plasmídeos , Antipirina/análogos & derivados , Antipirina/farmacologia , Benzimidazóis/farmacologia , Dimetil Sulfóxido/farmacologia , Raios gama , Radiossensibilizantes/farmacologiaRESUMO
To elucidate the kinetics of the induction of DNA strand breaks by low-energy Auger electron emitters, we compared the yields of DNA breaks in supercoiled pUC19 DNA after the decay of 125I (1) in proximity to DNA after minor-groove binding (125I-iodoHoechst 33342, 125IH) and (2) at a distance from DNA (125I-iodoantipyrine, 125IAP). Iodine-125 bound to the minor groove in DNA or free in solution is equally effective per decay in producing single-strand breaks (SSBs), while 125I bound to the minor groove is 6.7-fold more efficient than 125I free in solution in producing double-strand breaks (DSBs) (1.08 +/- 0.13 compared to 0.16 +/- 0.01 DSB/decay). Consequently, SSB to DSB ratios for 125IAP and gamma radiation (20.7 +/- 2.9 and 43.8 +/- 1.5, respectively) are greater than that for 125IH (2.9 +/- 0.4). Finally, the decay of 125IH leads to fragmentation of plasmid DNA beyond SSBs and DSBs.
Assuntos
Dano ao DNA , DNA Super-Helicoidal/efeitos da radiação , Radioisótopos do Iodo/química , Antipirina/análogos & derivados , Antipirina/síntese química , Antipirina/química , Benzimidazóis/síntese química , Benzimidazóis/química , DNA/química , DNA/efeitos da radiação , DNA Super-Helicoidal/química , Elétrons , Raios gama , Cinética , Plasmídeos/genética , Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/químicaRESUMO
PURPOSE: The incubation of the DNA minor-groove binder [125I]iodoHoechst 33342 (125IH) with plasmid DNA leads to the production of one double-strand break (dsb) per decay, both in the presence and absence of dimethylsulfoxide (DMSO). In contrast, when 125I is incorporated into mammalian cell DNA as an iodinated pyrimidine base, DMSO decreases the dsb yield and enhances survival. Because these variations in radioprotective effects may be due either to the location of 125I vis-à-vis the DNA helix or to differences in DNA architecture, the toxicity of 125IH and its modification by DMSO were examined in mammalian cells. METHODS: Uptake and retention of 125IH in V79 cells were measured, and survival was determined after accumulation of 125I decays at 0.3 degrees C +/-10% DMSO. RESULTS: A linear-quadratic survival curve was obtained both in the absence [D37 = 114+/-36 decays/cell, alpha = (5.39 1.17) x10(-3) cell/decay] and presence [D37 = 211+/-65 decays/cell, alpha = (1.27+/-0.52) x10(-3) cell/decay] of DMSO. The dose modification factor for the linear component of the survival curve was 4.25+/-1.97, indicating the predominance of indirect mechanisms. This value is similar to that obtained with DNA-incorporated 125I (4.05+/-1.72) and for the initial slope (alpha) of 137Cs gamma-rays (4.43+/- 1.41). CONCLUSIONS: Cytotoxicity resulting from the decay of the Auger electron emitter 125I in the mammalian cell nucleus is caused mainly by indirect mechanisms.
Assuntos
Benzimidazóis/toxicidade , Dimetil Sulfóxido/farmacologia , Sequestradores de Radicais Livres/farmacologia , Compostos Radiofarmacêuticos/toxicidade , Animais , Benzimidazóis/metabolismo , Benzimidazóis/farmacocinética , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Cricetinae , Cricetulus , DNA/metabolismo , DNA/efeitos da radiação , Dano ao DNA , Radioisótopos do Iodo/metabolismo , Radioisótopos do Iodo/toxicidade , Pulmão/citologia , Conformação de Ácido Nucleico , Compostos Radiofarmacêuticos/metabolismo , Compostos Radiofarmacêuticos/farmacocinéticaRESUMO
A new lipophilic fluorescein probe (fluor-DHPE) has been identified that can assay lipid peroxidation in mammalian cells on a cell-by-cell or selected-cell-subpopulation basis by flow cytometry. Application of this approach requires that the fluorescent probe be nonexchangeable among cells. Fluorescein is an appropriate fluorophore, since its fluorescence matches the specifications of common flow cytometers and the compound loses its fluorescence upon reaction with peroxyl radicals. Upon examination of four lipophilic derivatives of fluorescein, fluor-DHPE was found to be the only probe that was nonexchangeable among labeled and unlabeled rat RBC for at least 24 h. The exposure of fluor-DHPE-labeled RBC to benzoyl peroxide followed by mixing the sample with RBC unexposed to peroxide led to a decrease in fluorescence. Furthermore, the flow cytometer could clearly select the subpopulation of cells undergoing lipid peroxidation from those cells that were not. Fluor-DHPE-labeled-RBC obtained from rats and exposed to cumene hydroperoxide also displayed a gradual decrease in fluorescence. This decrease was preventable by either regulation of the vitamin E content in the animal diet or in vitro supplementation of cells with vitamin E. We conclude that fluor-DHPE is a stable and nonexchangeable probe for monitoring lipid peroxidation in cell subpopulations by flow cytometry.
Assuntos
Citometria de Fluxo/métodos , Fluoresceínas/metabolismo , Fosfatidiletanolaminas/metabolismo , Animais , Derivados de Benzeno/farmacologia , Peróxido de Benzoíla/metabolismo , Eritrócitos/metabolismo , Fluorescência , Corantes Fluorescentes/metabolismo , Peroxidação de Lipídeos/fisiologia , Masculino , Ratos , Ratos Endogâmicos F344RESUMO
Asynchronous Chinese hamster V79 lung fibroblasts were incubated at 37 degrees C for 30 min with the thymidine analog 5-[211At]astato-2'-deoxyuridine (211AtdU, exposure from DNA-incorporated activity) or with [211At]astatide (211At-, exposure from extracellular activity), and DNA-incorporated activity was determined. The 211AtdU content in cellular DNA increased as a function of extracellular concentration. Incorporation of 211At- was less than 1% of that of 211AtdU. After exposure, cells were frozen in the presence of 10% DMSO. One month later, survival was determined by the colony-forming assay, and DNA double-strand breaks (DSBs) were measured by the neutral elution method (pH 9.6). The survival curve for 211AtdU was biphasic (D37 = 2.8 decays per cell), reflecting killing of 211At-DNA-labeled cells and of unlabeled cells irradiated by 211At in neighboring labeled cells. The toxicity of 211At- decaying outside the cell (30-min exposure) was negligible. Analysis of the survival curve produced a D0 of 1.3 decays/cell for 211At-labeled cells. The yield of DSBs from the decay of DNA-incorporated 211At was compared with that from DNA-incorporated 125I. Each decay of 211At produced at least 10 times the number of DSBs as that obtained per 125I decay. The extreme radiotoxicity of DNA-incorporated 211AtdU seems to be associated with considerable damage to the mammalian cell genome.
Assuntos
Astato/farmacocinética , Dano ao DNA , Fibroblastos/efeitos da radiação , Idoxuridina/análogos & derivados , Animais , Sobrevivência Celular/efeitos da radiação , Células Cultivadas/metabolismo , Células Cultivadas/efeitos da radiação , Ensaio de Unidades Formadoras de Colônias , Cricetinae , Cricetulus , DNA/metabolismo , DNA/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/metabolismo , Idoxuridina/farmacocinéticaRESUMO
UNLABELLED: Glial neoplasms of the human central nervous system have defied treatment, in part because of the limited selectivity of available cytotoxic agents. The thymidine analog 5-iodo-2'-deoxyuridine radiolabeled with the Auger electron emitter 125I (125IUdR) is highly toxic to dividing cells when it is deoxyribonucleic acid incorporated, but it is relatively innocuous when located outside the nucleus. Previous studies have shown that 125IUdR has significant antineoplastic potential against mammalian cells in vitro and direct administration of 125IUdR is effective therapy for ovarian ascites tumors in mice and neoplastic meningitis in rats. Studies using external gamma imaging and autoradiography have also shown that direct intratumoral administration of 123IUdR/125IUdR into intracerebral 9L gliosarcomas in rats results in selective uptake of the radionuclide into tumor cells. Based on these encouraging results, we have evaluated the therapeutic potential of 125IUdR in rats bearing intracerebral 9L gliosarcomas. METHODS: Iodine-125-IUdR was infused intracerebrally over a 2-day period into rats bearing 1-day-old 9L tumors and over a 6-day period into animals with 9-day-old 9L tumors; equimolar concentrations of 127IUdR were infused into control animals. Tumor growth was monitored by contrast-enhanced 1H MRI and animal survival was followed over time. RESULTS: Intracerebral tumors (3-7 mm) were readily detected by MRI. Tumor-bearing rats treated with 127IUdR succumbed within 17-24 days, whereas tumor-bearing animals treated with 125IUdR survived significantly longer, and 10%-20% of the animals were cured of tumors. CONCLUSION: These data substantiate the antineoplastic potential of 5-[125I]iodo-2'-deoxyuridine and indicate that it may be a useful agent for the therapy of solid tumors that are accessible to direct radiopharmaceutical administration.
Assuntos
Neoplasias Encefálicas/radioterapia , DNA/biossíntese , Gliossarcoma/radioterapia , Idoxuridina/uso terapêutico , Radioisótopos do Iodo/uso terapêutico , Inibidores da Síntese de Ácido Nucleico/uso terapêutico , Animais , Antineoplásicos/administração & dosagem , Antineoplásicos/uso terapêutico , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Gliossarcoma/patologia , Idoxuridina/administração & dosagem , Injeções Intralesionais , Radioisótopos do Iodo/administração & dosagem , Imageamento por Ressonância Magnética , Transplante de Neoplasias , Inibidores da Síntese de Ácido Nucleico/administração & dosagem , Compostos Radiofarmacêuticos/administração & dosagem , Compostos Radiofarmacêuticos/uso terapêutico , Ratos , Ratos Endogâmicos F344 , Técnicas Estereotáxicas , Fatores de TempoRESUMO
Ethidium homodimer (EHD) was conjugated to B72.3 monoclonal antibody using a method whereby 85-90% of the conjugated EHD remains available for DNA intercalation. Antibody was thiopropionylated by reaction with N-succinimidyl 3-(2-pyridyldithio)propionate and reduction of pyridyldithio groups with dithiothreitol. EHD was maleimido-functionalized with succinimidyl-4-(N-maleimidoethyl)cyclohexane-1-carboxylate and treated with thiopropionylated antibody to obtain a conjugate containing approximately 3.4 EHD per antibody molecule. For biologic studies, 14C-labeled EHD was synthesized by reductive amination and conjugated as above. In vitro the conjugate maintained chemical integrity and immunoreactivity, while in vivo its targeting of LS174T tumors was reduced compared with that of iodinated antibody. A decrease in isoelectric point of the immunoconjugate was also observed.
Assuntos
Adenocarcinoma/metabolismo , Anticorpos Monoclonais/metabolismo , Anticorpos Antineoplásicos/metabolismo , Radioisótopos de Carbono/farmacocinética , Neoplasias do Colo/metabolismo , Etídio/análogos & derivados , Radioisótopos do Iodo/farmacocinética , Animais , Dimerização , Humanos , Indicadores e Reagentes , Lisina , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Distribuição TecidualRESUMO
The synthesis of halogenated rhodamine (Rh) derivatives was carried out by controlling the stoichiometry of the halogenating agents, bromine and iodine monochloride. In the no-carrier-added synthesis of radioiodinated rhodamine 123, direct labeling of rhodamine 123 (Rh 123) with Na125I/Na131I required the presence of the oxidant peracetic acid. 125I/131I-Rh 123 was synthesized in modest yields (40-45%). HPLC purification separated Rh 123 from its mono- and diiodo derivatives. Monohalogenation of Rh 123 did not alter the compound's ability to permeate viable cells and localize in mitochondria. 125I/131I-Rh 123 was stable in serum in vitro but rapidly metabolized after intravenous injection into mice. Consequently, scintigraphy and biodistribution data reveal poor targeting of subcutaneously growing human tumor xenografts. The results are compared to those obtained following the administration of [99mTc]hexakis(2-methoxyisobutylisonitrile) which also did not image human tumor xenografts in nude mice.
Assuntos
Compostos Radiofarmacêuticos/síntese química , Compostos Radiofarmacêuticos/farmacocinética , Tecnécio Tc 99m Sestamibi/farmacocinética , Animais , Carcinoma de Células Escamosas/diagnóstico por imagem , Cromatografia Líquida de Alta Pressão , Humanos , Marcação por Isótopo , Células KB , Camundongos , Camundongos Nus , Microscopia de Fluorescência , Neoplasias Nasofaríngeas/diagnóstico por imagem , Transplante de Neoplasias , Cintilografia , Rodaminas/síntese química , Rodaminas/farmacocinética , Distribuição Tecidual , Transplante Heterólogo , Células Tumorais CultivadasRESUMO
We have examined whether nuclear DNA can be protected from double-strand breaks (DSBs) induced by decay of the Auger-electron-emitting radionuclide 125I. Decays were accumulated at 0.3 degrees C in Chinese hamster V79 cells suspended in isotonic buffer containing 0.1 M EDTA in the presence or absence of 10% dimethyl sulfoxide (DMSO). DSBs were measured by the neutral elution method (pH 9.6) and quantified as strand scission factors. DMSO was shown to protect DNA from DSBs caused by the decay of DNA-incorporated 125I. The dose modification factor (DMF) for this radionuclide decreases as a function of 125I decays (389 to 4,100 decays, DMF = 2.5 to 1.3). Extrapolation of the curve for the DMF indicates that at approximately 15,000 decays/cell, a DMF of 1 would be obtained. Experiments using large numbers of 125I decays confirmed these extrapolations. For induction of DSBs by 137Cs gamma rays, the DMF also decreases with dose (50 to 290 Gy, DMF = 2.7 to 1.5). However, extrapolation of the curve for the DMF indicates that protection does not cease at higher doses. The data show that, at the same level of damage, DMSO can protect against gamma-ray-induced DSBs 1.35-fold more efficiently than against DSBs caused by the decay of DNA-incorporated 125I. It appears that when 125I is incorporated into DNA, chromatin structure fosters some DSB formation by an indirect mechanism(s) and that more than one DSB is generated per decaying atom.
Assuntos
Dano ao DNA/efeitos da radiação , DNA/efeitos da radiação , Radioisótopos do Iodo , Animais , Células Cultivadas , Cricetinae , Cricetulus , Fragmentação do DNA , Dimetil Sulfóxido/farmacologia , Sequestradores de Radicais Livres , Raios gama , Hidróxidos , Nucleossomos/efeitos da radiaçãoRESUMO
We have examined whether mammalian cells in vitro can be protected against the lethal effects of irradiation by Auger electrons emitted from DNA-incorporated 125I. Chinese hamster V79 lung fibroblasts were cultivated in the presence of 5-[125I]iodo-2'-deoxyuridine (125IdU) for 18 h and resuspended in ice-cold medium in the presence or absence of 10% dimethyl sulfoxide (DMSO). DNA-incorporated 125I activity was measured and the cells were plated for survival. A portion of the cell suspensions were also stored on ice to accumulate 125I decays for 6 to 48 h, after which the cells were plated to determine survival. Storage on ice up to 48 h without radioactivity reduced plating efficiency from 67 +/- 4% (SEM) to 20 +/- 1%. DMSO had a protective effect on colony formation, as the respective cloning efficiencies were 83 +/- 3% and 72 +/- 12% at 0 and 48 h. The survival curves for 125IdU-labeled cells are exponential with D0 = 36 +/- 2 decays per cell in the absence of DMSO and 195 +/- 20 decays per cell in the presence of DMSO. Thus the dose modification factor (DMF) at 37% survival for 10% DMSO is 5.4 +/- 0.6 for DNA-incorporated 125I. In reference experiments, a DMF of 2.5 +/- 0.8 was measured for cells irradiated with 137Cs gamma rays. These results indicate that the radiotoxicity of Auger electrons from 125I decay in mammalian cells is caused mainly by an indirect mechanism(s).