18F-FLT and 125I-IdUrd uptake increase in human tumour cell lines induced by the thymidylate synthase inhibitor FdUrd.

AIM: 5-fluoro-2'-deoxyuridine (FdUrd) depletes the endogenous 5'-deoxythymidine triphosphate (dTTP) pool. We hypothesized whether uptake of exogenous dThd analogues could be favoured through a feedback enhanced salvage pathway and studied the FdUrd effect on cellular uptake of 3'-deoxy-3'-18F-fluorothymidine (18F-FLT) and 5-125I-iodo-2'-deoxyuridine (125I-IdUrd) in different cancer cell lines in parallel. METHODS: Cell uptake of 18F-FLT and 125I-IdUrd was studied in 2 human breast, 2 colon cancer and 2 glioblastoma lines. Cells were incubated with/without 1 µmol/l FdUrd for 1 h and, after washing, with 1.2 MBq 18F-FLT or 125I-IdUrd for 0.3 to 2 h. Cell bound 18F-FLT and 125I-IdUrd was counted and expressed in % incubated activity (%IA). Kinetics of 18F-FLT cell uptake and release were studied with/without FdUrd modulation. 2'-3H-methyl-fluorothymidine (2'-3H-FLT) uptake with/without FdUrd pretreatment was tested on U87 spheroids and monolayer cells. RESULTS: Basal uptake at 2 h of 18F-FLT and 125I-IdUrd was in the range of 0.8-1.0 and 0.4-0.6 Bq/cell, respectively. FdUrd pretreatment enhanced 18F-FLT and 125I-IdUrd uptake 1.2-2.1 and 1.7-4.4 fold, respectively, while co-incubation with excess thymidine abrogated all 18F-FLT uptake. FdUrd enhanced 18F-FLT cellular inflow in 2 breast cancer lines by factors of 1.8 and 1.6, respectively, while outflow persisted at a slightly lower rate. 2'-3H-FLT basal uptake was very low while uptake increase after FdUrd was similar in U87 monolayer cells and spheroids. CONCLUSIONS: Basal uptake of 18F-FLT was frequently higher than that of 125I-IdUrd but FdUrd induced uptake enhancement was stronger for 125I-IdUrd in five of six cell lines. 18F-FLT outflow from cells might be an explanation for the observed difference with 125I-IdUrd.

Fluorodeoxyuridine mediated cell cycle synchronization in S-phase increases the Auger radiation cell killing with 125I-iododeoxyuridine.

AIM: 125I-iododeoxyuridine is a potential Auger radiation therapy agent. Its incorporation in DNA of proliferating cells is enhanced by fluorodeoxyuridine. Here, we evaluated therapeutic activities of 125I-iododeoxyuridine in an optimized fluorodeoxyuridine pre-treatment inducing S-phase synchronization. METHODS: After S-phase synchronization by fluorodeoxyuridine, cells were treated with 125I-iododeoxyuridine. Apoptosis analysis and S-phase synchronization were studied by flow cytometry. Cell survival was determined by colony-forming assay. Based on measured growth parameters, the number of decays per cell that induced killing was extrapolated. RESULTS: Treatment experiments showed that 72 to 91% of synchronized cells were killed after 0.8 and 8 kBq/ml 125I-iododeoxyuridine incubation, respectively. In controls, only 8 to 38% of cells were killed by corresponding 125I-iododeoxyuridine activities alone and even increasing the activity to 80 kBq/ml gave only 42 % killing. Duplicated treatment cycles or repeated fluorodeoxyuridine pre-treatment allowed enhancing cell killing to >95 % at 8 kBq/ml 125I-iododeoxyuridine. About 50 and 160 decays per S-phase cells in controls and S-phase synchronization, respectively, were responsible for the observed cell killing at 0.8 kBq/ml radio-iododeoxyuridine. CONCLUSION: These data show the successful application of fluorodeoxyuridine that provided increased 125I-iododeoxyuridine Auger radiation cell killing efficacy through S-phase synchronization and high DNA incorporation of radio-iododeoxyuridine.

A balanced deoxyribonucleoside mixture increased the rate of DNA incorporation of 5-[125I]Iodo-2'-deoxyuridine in glioblastoma cells.

Administration of nucleotide mixtures has been shown to restore and sustain the proliferation of leukocytes and enterocytes. Since it has been suggested that cancer cells use exogenous nucleotides more efficiently than normal cells, we hypothesized that administration of nucleotide mixtures would also stimulate the proliferation of cancer cells, thereby increasing the number of cells targeted by the thymidine analog 5-[(125)I]iodo-2'-deoxyuridine ([(125)I]IUdR). We first evaluated the influence of different deoxyribonucleoside mixtures on the DNA incorporation of [(125)I]IUdR in 3 human glioblastoma cell lines. Results showed that a 4-h coincubation with a mixture of identical concentration (10 microM) of deoxyadenosine, deoxyuridine, deoxyguanosine and deoxycytidine (AUGC) increased by 8.5-, 6.2-, and 2.0-fold the rate of DNA incorporation of [(125)I]IUdR in exponentially growing LN229, U87 and U251 cells, respectively. Replacing deoxyuridine by thymidine (ATGC) reversed the effect of the mixture, whereas removing deoxyuridine allowed a mixture of 10 microM AGC to increase by 2.2-fold the rate of DNA incorporation of [(125)I]IUdR in LN229 cells. Furthermore, the rate of DNA incorporation of [(125)I]IUdR in LN229 and U87 cells was increased up to 19.9- and 9.4-fold, respectively, by extending the coincubation time with 10 microM AUGC to 9 h, and up to 40.9- and 26.8-fold by incubating confluent cells for 4 h with 10 microM AUGC. Flow cytometry analysis showed that exposure of confluent cells to AUGC increased the percentage of cells in S phase of the cell cycle. Thus, co-administration of a balanced deoxyribonucleoside mixture may improve the use of radiolabeled nucleotide analogs, such as [(125)I]IUdR, for the targeting of cancer cells.

Food intake in 1707 hospitalised patients: a prospective comprehensive hospital survey.

BACKGROUND & AIMS: This study aimed to assess the ability of the hospital meal service to meet patients' nutritional needs. METHODS: All hospitalised patients who received 3 meals/day without artificial nutritional support were included. The nutritional values of food served, consumed and wasted during a 24 h period were compared to patients' needs estimated as energy: 110% Harris-Benedict formula; protein: 1.2 or 1.0 g/kg bodyweight/day for patients < or = or > 65 years old, respectively. A structured interview recorded patients' evaluation of the meal quality, their reasons for non-consumption of food and the relationship between food intake and disease. RESULTS: Out of 1707 patients included, 1416 were fully assessable (59% women; 68+/-21 years; body mass index: 24.3+/-5.1 kg/m(2)). Daily meals provided 2007+/-479 kcal and 78+/-21 g of protein and exceeded patients' needs by 41% and 15%, respectively. However, 975 patients did not eat enough. Plate waste was 471+/-372 kcal and 21+/-17 g of protein/day/patient. Moreover, the food intake of 572 (59%) of these underfed patients was not predominantly affected by disease. Logistic regression analyses identified as other risk factors: elevated BMI, male gender, modified diet prescription, length of stay <8 or > or = 90 days and inadequate supper. CONCLUSION: Despite sufficient food provision, most of the hospitalised patients did not cover their estimated needs. Since insufficient food intake was often attributed to causes other than disease, there should be potential to improve the hospital meal service.

[Early detection of protein-calorie malnutrition: a therapeutic asset]. / Détection précoce de la malnutrition protéino-calorique: un atout thérapeutique.

In the industrialised countries, the sedentary life and the ageing process of the population, as well as the more frequent chronic diseases and heavy treatments, increase the incidence of protein-energy malnutrition (PEM). The insidious nature and harmful outcome of the PEM on the recovery process requires careful attention of the practitioner to the clinical signs of PEM. Their detection includes an anamnesis, anthropometric examinations, and assessments of the nutritional intakes and the impact of disease and medico-surgical treatments. However, the loss of muscle mass, which is the main indicator of the PEM, is often only assessed by the measurement of the body composition using bioelectrical impedance analysis. The advantage of this method is to distinguish fat-free mass, including muscle mass, from fat mass, when the loss of muscle is hidden by an increase of fat mass and/or body water. Using these different tools allows the practitioner to early detect PEM, to identify its causes, and to establish an appropriate nutritional schedule, in order to prevent from PEM or correct it.

Fluorodeoxyuridine improves imaging of human glioblastoma xenografts with radiolabeled iododeoxyuridine.

Use of radiolabeled nucleotides for tumor imaging is hampered by rapid in vivo degradation and low DNA-incorporation rates. We evaluated whether blocking of thymidine (dThd) synthesis by 5-fluoro-2'-deoxyuridine (FdUrd) could improve scintigraphy with radio-dThd analogues, such as 5-iodo-2'-deoxyuridine (IdUrd). We first show in vitro that coincubation with FdUrd substantially increased incorporation of [125I]IdUrd and [3H]dThd in the three tested human glioblastoma lines. Flow cytometry analysis showed that a short coincubation with FdUrd (1 h) produces a signal increase per labeled cell. We then measured biodistribution 24 h after i.v. injection of [125I]IdUrd in nude mice s.c. xenografted with the three glioblastoma lines. Compared with animals given [125I]IdUrd alone, i.v. preadministration for 1 h of 10 mg/kg FdUrd increased the uptake of [125I]IdUrd in the three tumors 4.8-6.8-fold. Compatible with previous reports, there were no side effects in mice observed for 2 months after receiving such a treatment. The tumor uptake of [125I]IdUrd was increased < or =13.6-fold when FdUrd preadministration was stepwise reduced to 1.1 mg/kg. Uptake increases remained lower (between 1.7- and 5.8-fold) in normal proliferating tissues (i.e., bone marrow, spleen, and intestine) and negligible in quiescent tissues. DNA extraction showed that 72-80% of radioactivity in tumor and intestine was bound to DNA. Scintigraphy of xenografted mice was performed at different times after i.v. injection of 3.7 MBq [125I]IdUrd. Tumor detection was significantly improved after FdUrd preadministration while still equivocal after 24 h in mice given [125I]IdUrd alone. Furthermore, background activity could be greatly reduced by p.o. administration of KClO4 in addition to potassium iodide. We conclude that FdUrd preadministration may improve positron or single photon emission tomography with cell division tracers, such as radio-IdUrd and possibly other dThd analogues.

Unlabelled iododeoxyuridine increases the cytotoxicity and incorporation of [1251]-iododeoxyuridine in two human glioblastoma cell lines.

Iododeoxyuridine (IUdR), labelled with radioiodines emitting Auger, alpha or beta- radiation, has been proposed as a therapeutic tool in the treatment of cancer. However, the low per cent incorporation in tumour cells and limited cytotoxicity are major obstacles for such an application. Using unlabelled IUdR as a modulator, we have studied the in vitro cytotoxicity of [125I]-IUdR in two human glioblastoma cell lines. Surprisingly, an enhanced cytotoxicity of [125I]-IUdR was observed in the presence of 0.3-10 microM concentrations of unlabelled IUdR in U251 glioblastoma cells and to a lesser extent in LN229 cells. The presence of unlabelled IUdR unexpectedly increased the incorporation of [125I]-IUdR in both cell lines. Thymidine competitively blocked the cytotoxic effects of combined unlabelled and [125I]-labelled IUdR in these cells and DNA-incorporation of radiolabelled IUdR.

Combined radioimmunotherapy and radiotherapy of liver metastases from colorectal cancer: a feasibility study.

BACKGROUND: A combination of radioimmunotherapy (RIT) and radiotherapy (RT) should allow one to increase the dose of radiation targeting a particular tumour without the concomitant increase of toxic side effects. This might be obtained if the dose limiting side effect of each individual radiation therapy concerned different organs. METHODS: Six patients with limited liver metastatic disease from colorectal cancer were treated with 6.9 GBq (range 4.7 to 8.4 GBq) 131I-labelled anti-CEA MAb F(ab')2 fragments combined with 20 Gy RT to the liver. Both treatments were given in close association, according to timing schedules evaluated in animals that gave the best results. RESULTS: Reversible bone marrow and liver toxicity was observed in 6 and 5 patients, respectively. Three patients who first received 20 Gy RT to the liver, showed a significant platelet drop upon completion of RT. Repeat computerized tomography (CT) after 2 months showed a minor response in 1 patient and stable disease in 3 patients. CONCLUSION: The study shows potential ways of combining RIT and RT, suggesting that this combination is feasible for the treatment of liver metastases.

Radioimmunotherapy of colorectal cancer liver metastases: combination with radiotherapy.

The expected therapeutic gain of a combined radioimmunotherapy (RIT) with conventional radiotherapy (RT) would be a synergy of tumor irradiation, provided that toxic, dose-limiting side effects concern different organs. We have shown in a model of subcutaneous human colon cancer transplants in nude mice that RIT with 131I-labeled anti-CEA antibody fragments combined with fractionated RT give an additive therapeutic effect without increase of side effects. A second study of different timing schedules of RIT and RT has shown that close association of both therapies without delay is more efficient than a therapy with a treatment-free interval of two weeks. In a new model of human colon cancer liver metastases in nude mice, early treatment with RIT and with RT has been curative, whereas therapies initiated later were less efficient, suggesting that the combined therapy is likely to be more efficient in an adjuvant situation after surgery. At the clinical level, six patients with limited liver metastatic disease from colorectal cancer were treated with RIT using 200 mCi 131I-labeled anti-CEA MAb F(ab')2 fragments combined with fractionated external beam RT of 20 Gy to the entire liver. As expected, spontaneously reversible bone marrow toxicity grade 3 to 4 and reversible liver toxicity grade 1 to 3 have been observed. By computerized tomography, three patients showed stable disease and one patient partial remission, whereas two patients had progressive disease. In conclusion, animal experiments have shown a clear advantage of combined RT and RIT, and the clinical study shows the feasibility of such a therapy in patients with colorectal cancer liver metastases.