Autoradiographic and histopathological studies of boric acid-mediated BNCT in hepatic VX2 tumor-bearing rabbits: Specific boron retention and damage in tumor and tumor vessels.

Hepatoma is a malignant tumor that responds poorly to conventional therapies. Boron neutron capture therapy (BNCT) may provide a better way for hepatoma therapy. In this research, (10)B-enriched boric acid (BA, 99% (10)B) was used as the boron drug. A multifocal hepatic VX2 tumor-bearing rabbit model was used to study the mechanisms of BA-mediated BNCT. Autoradiography demonstrated that BA was selectively targeted to tumors and tumor vessels. Histopathological examination revealed the radiation damage to tumor-bearing liver was concentrated in the tumor regions during BNCT treatment. The selective killing of tumor cells and the destruction of the blood vessels in tumor masses may be responsible for the success of BA-mediated BNCT for liver tumors.

BNCT for locally recurrent head and neck cancer: preliminary clinical experience from a phase I/II trial at Tsing Hua Open-Pool Reactor.

To introduce our preliminary experience of treating locally and regionally recurrent Head and Neck cancer patients at Tsing Hua Open-Pool Reactor in Taiwan, four patients (M/F=3/1, median age 68 Y/O) were enrolled. BNCT with BPA (400 mg/kg) injected in 2 phases and prescription dose of 12-35 Gy (Eq.)/fraction for 2 fractions at 30 day interval can be given with sustained blood boron concentration and tolerable early toxicities for recurrent H & N cancer.

Low dose of gamma irradiation enhanced boronophenylalanine uptake in head and neck carcinoma cells for boron neutron capture therapy.

This study attempted to increase the boron uptake of human head and neck carcinoma SAS cells for BNCT by using a gamma dose of 0.1 Gy for combined treatment. Intracellular boron concentrations in 25 µgB/mL medium of BPA treated and BPA combined gamma-irradiation treated SAS cells were 73.8±1.73 and 95.15±1.36 ppm, respectively. After neutron irradiation, the G2/M-phase cell populations of untreated, BPA treated and BPA combined gamma-irradiation treated SAS cells were 19.31±1.71%, 52.47±2.25% and 59.19±2.63%, respectively. Experimental results indicate that the low dose gamma radiation with combination BPA treatment has the highest killing rate after neutron irradiation. Capable of significantly increasing the G2/M arrest after neutron irradiation, the combined treatment of a low dose of gamma irradiation with 25 µg B/mL medium of BPA also provided a higher killing effect for BNCT.

Potential of using boric acid as a boron drug for boron neutron capture therapy for osteosarcoma.

Osteosarcoma is a malignant tumor commonly found in human and animals. The ability of boric acid (BA) to accumulate in osteosarcoma due to the mechanism of the bone formation of cancer cells would make boron neutron capture therapy (BNCT) an alternative therapy for osteosarcoma. This study evaluated the feasibility of using BA as the boron drug for BNCT of bone cancer. The cytotoxicity of BA to L929 cells exceeded that of UMR-106 cells. With 25 µg (10)B/mL medium of BA treatment, the boron concentration in UMR-106 cells was higher than that in L929 cells. The biodistribution and pharmacokinetics of BA in Sprague-Dawley (SD) rats were studied by administrating 25 mg (10)B/kg body weight to SD rats. Blood boron level decreased rapidly within one hour after BA injection. Boron concentration in the long bone was 4-6 time higher than that of blood. Results of this study suggest that BA may be a potential drug for BNCT for osteosarcoma.

Biodistribution of phenylboric acid derivative entrapped lipiodol and 4-borono-2-18F-fluoro-L-phenylalanine-fructose in GP7TB liver tumor bearing rats for BNCT.

A new phenylboric acid derivative entrapped lipiodol (PBAD-lipiodol) was developed as a boron carrier for the boron neutron capture therapy (BNCT) of hepatoma in Taiwan. The biodistribution of both PBAD-lipiodol and BPA-fructose was assayed in GP7TB hepatoma-bearing rat model. The highest uptake of PBAD-lipiodol was found at 2h post injection. The application of BNCT for the hepatoma treatment in tumor-bearing rats is suggested to be 2-4h post PBAD-lipiodol injection.

Microdosimetry study of THOR BNCT beam using tissue equivalent proportional counter.

Boron neutron capture therapy (BNCT) is a cancer treatment modality using a nuclear reactor and a boron compound drug. In Taiwan, Tsing Hua open-pool reactor (THOR) has been modulated for the basic research of BNCT for years. A new BNCT beam port was built in 2004 and used to prepare the first clinical trial in the near future. This work reports the microdosimetry study of the THOR BNCT beam by means of the tissue equivalent proportional counter (TEPC). Two self-fabricated TEPCs (the boron-doped versus the boron-free counter wall) were introduced. These dual TEPCs were applied to measure the lineal energy distributions in air and water phantom irradiated by the THOR BNCT mixed radiation field. Dose contributions from component radiations of different linear energy transfers (LETs) were analyzed. Applying a lineal energy dependent biological weighting function, r(y), to the total and individual lineal energy distributions, the effective relative biological effectiveness (RBE), neutron RBE, photon RBE, and boron capture RBE (BNC RBE) were all determined at various depths of the water phantom. Minimum and maximum values of the effective RBE were 1.68 and 2.93, respectively. The maximum effective RBE occurred at 2cm depth in the phantom. The average neutron RBE, photon RBE, and BNC RBE values were 3.160+/-0.020, 1.018+/-0.001, and 1.570+/-0.270, respectively, for the THOR BNCT beam.

Suitability of boron carriers for BNCT: accumulation of boron in malignant and normal liver cells after treatment with BPA, BSH and BA.

Hepatocellular carcinoma remains widely prevalent in tropical Africa and south-east Asia. At present, there are no effective treatments for hepatoma and its prognosis is extremely poor unless the tumor was diagnosed in an early stage and resected before metastasis. Therefore, boron neutron capture therapy (BNCT) may provide an alternative therapy for treatment of hepatocellular carcinoma. In this study, the intracellular concentrations of L-boronophenylalanine (BPA), sodium borocaptate (BSH) and boric acid (BA) were examined in human hepatoma HepG2 and liver Clone 9 cell cultures. With the use of 25 microgB/mL media of BPA, BSH and BA, the intracellular uptake of boron in HepG2 and Clone 9 cells was compared. The suitability of BPA, BSH and BA were further evaluated on the basis of organ-specific boron distribution in normal rat tissues. BPA, BSH and BA were administered via intraperitoneal injection into rats with corresponding boron concentrations of 7, 25, and 25mg/kg body weight, respectively. The accumulation rates of BPA, BSH and BA in HepG2 cells were higher than that of Clone 9 cells. Boron concentration in BPA, BSH and BA treated HepG2 cells were 1.8, 1.5, and 1.6-fold of Clone 9 cells at 4h, respectively. In both HepG2 and Clone 9 cells, although the concentration of boron in BPA-treated cells exceeded that in BA-treated ones, however, cells treated with BPA had similar surviving fraction as those treated with BA after neutron irradiation. The accumulation ratios of boron in liver, pancreas and kidney to boron in blood were 0.83, 4.16 and 2.47, respectively, in BPA treated rats, and 0.75, 0.35 and 2.89, respectively, in BSH treated rats at 3h after treatment. However, boron does not appear to accumulate specifically in soft tissues in BA treated rats. For in situ BNCT of hepatoma, normal organs with high boron concentration and adjacent to liver may be damaged in neutron irradiation. BPA showed high retention in pancreas and may not be a good drug for BNCT of hepatoma. BSH had higher retention in liver but low level in pancreas and spleen appears to be a better candidate BNCT drug for hepatoma. These preliminary results provide useful information on future application of BNCT for hepatoma.

Assessment of dose rate scaling factors used in NCTPlan treatment planning code for the BNCT beam of THOR.

Tsing Hua open-pool reactor (THOR) at Tsing Hua University in Taiwan has been used to investigate the feasibility and to enhance the technology of boron neutron capture therapy (BNCT) for years. A rebuilt epithermal beam port for BNCT at THOR was finished in the summer of 2004, and then researches and experiments were performed to hasten the first clinical treatment case of BNCT in Taiwan in the near future. NCTPlan, a Monte Carlo-based clinical treatment planning code, was used to calculate the dose-rate distributions of BNCT in this work. A self-made Snyder head phantom with a servo-motor control system was irradiated in front of the THOR BNCT beam exit. The phantom was made from a 3mm shell of quartz wool impregnated with acrylic casting resin mounted on an acrylic base, and was filled with water. Gold foils (bare and cadmium-covered) and paired ion chambers (one with graphite wall and filled with CO(2) gas, another with A-150 plastic tissue equivalent wall and filled with tissue equivalent gas) were placed inside the Snyder phantom to measure and estimate the depth-dose distributions in the central axis of the beam. Dose components include the contribution of thermal neutrons, fast neutrons, photons and emitted alpha particles from (10)B(n,alpha)(7)Li reaction. Comparison and analysis between computed and measured results of depth-dose distributions were made in this work. Dose rate scaling factors (DRSFs) were defined as normalization factors derived individually for each dose component in the BNCT in-phantom radiation field that provide the best agreement between measured and computed data. This paper reports the in-phantom calculated and experimental dosimetry and the determined DRSFs used in NCTPlan code for the BNCT beam of THOR.

Morphology and function of gill mitochondria-rich cells in fish acclimated to different environments.

The objective of this study is to test the hypothesis that morphologically different mitochondria-rich (MR) cells may be responsible for the uptake of different ions in freshwater-adapted fish. Tilapia (Oreochromis mossambicus) were acclimated to high-Ca, mid-Ca, low-Ca, and low-NaCl artificial freshwater, respectively, for 2 wk. Cell densities of wavy-convex, shallow-basin, and deep-hole types of gill MR cells as well as whole-body Ca(2+), Na(+), and Cl(-) influxes were measured. Low-Ca fish developed more shallow-basin MR cells in the gills and a higher Ca(2+) influx than those acclimated to other media. However, fish acclimated to low-NaCl artificial freshwater predominantly developed wavy-convex cells, and this was accompanied by the highest Na(+) and Cl(-) influxes. Relative abundance of shallow-basin and wavy-convex MR cells appear to be associated with changes in Ca(2+) and Na(+)/Cl(-) influxes, suggesting that shallow-basin and wavy-convex MR cells are mainly responsible for the uptake of Ca(2+) and Na(+)/Cl(-), respectively.

Preparation and in vitro evaluation of B-lipiodol as a boron delivery agent for neutron capture therapy of hepatoma.

BACKGROUND: We prepared boron containing lipiodol (B-lipiodol), elucidated the retention of B-lipiodol in hepatoma cells and evaluated the in vitro cellular toxicity of B-lipiodol for neutron capture therapy. MATERIALS AND METHODS: Human hepatoma HepG2 cells were used to examine the uptake and retention of B-lipiodol. Light microscopes were used to examine the interaction and retention of B-lipiodol globules in individual hepatoma cells. Boron and lipiodol concentrations were determined by inductively coupled plasma-atomic emission spectroscopy and neutron activation analysis, respectively. RESULTS: The boron concentration in B-lipiodol drug could reach 2500 ppm. B-lipiodol could be stably retained in serum and culture medium. HepG2 cells appeared proficiently at internalization and persistent retention of B-lipiodol. The boron concentration reached 3.5 micrograms/10(6) cells without approaching saturation at 48 h treatment. CONCLUSION: Hepatoma cells could actively uptake B-lipiodol and a sufficient amount of boron was retained inside the HepG2 cells which could be used for neutron capture therapy.

Lipiodol uptake and retention by human hepatoma cells.

Lipodol has important diagnostic and therapeutic uses in hepatoma. However, the mechanisms of its selective, prolonged retention in hepatoma cells is not well understood. Therefore, using oil-red O, light and electron microscopy and neutron activation analysis we have determined that HepG2 cells are characterized by lipiodol deposition and emulsification on the cell surface, action uptake of lipodol by endocytosis, and prolonged intracellular retention. These findings may have major clinical significance in the development of a new treatment for hepatoma patients.

I-131-lipiodol cytotoxicity in hepatoma cells.

Hepatoma is a common cancer in Taiwan. New effective treatment for hepatoma patients is urgently needed. Encouraging results of I-131-lipiodol treatment for hepatoma with minimal toxicity have been recently reported. The mechanism of lipiodol targeting and retention by hepatoma are not well understood. The cellular interaction of lipiodol and the cytotoxic effects of I-131-lipiodol on hepatoma cells were investigated in this study. HepG2 cells were cultured with lipiodol, and untreated HepG2 cells were used as the control. Changes of cellular morphology were accessed by light and electron microscopy. The uptake and retention of lipiodol by HepG2 cells were studied by phase contrast microscopy and neutron activation analysis. HepG2 cells were cultured with I-131-lipiodol varying from 0.12 microCi to 120 microCi. The cytotoxic effect of I-131-lipiodol was evaluated by the surviving fraction of HepG2 cells. Changes in cellular morphology was examined by light microscopy. Results indicated that HepG2 cells were capable of active uptake of large amounts of lipiodol by endocytosis and prolonged intra-cellular retention associated with the formation of many bulging cytoplasmic extensions. I-131-lipiodol was highly cytotoxic to HepG2 cells. There was a steep dose response relationship, and the effective dose (LD50) was 1.2 microCi (480 rads). The cytotoxic effects of I-131-lipiodol were associated with pleomorphism of HepG2 cells, an increase in cell size and nuclear-cytoplasmic ratio, an increase in the size and number of nuclei, and vacuolation of the cytoplasm around the nuclear regions. Multiple nucleoli, fragmentation and segregation and ring shaped changes of nucleoli were also observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Simplified measurement of protein-bound iodine with epithermal neutron activation analysis.

A refined method for the measurement of protein-bound iodine (PBI) in blood serum was demonstrated by the use of epithermal neutron activation analysis (ENAA). PBI in ammonium sulfate-precipitated serum protein, after epithermal neutron activation, was determined by high resolution gamma-ray spectrometry. From our results, the PBI concentration was 67.8 +/- 2.2 ng/mL. Good agreement was obtained with published data, ranging from 40 to 80 ng/mL, which had been obtained using different analytical techniques. The validity of these techniques for PBI has been born out by a very good accuracy and simplicity without temperature dependence.

Salt-mediated multicell formation in Deinococcus radiodurans.

The highly radiation-resistant tetracoccal bacterium Deinococcus radiodurans exhibited a reversible multi-cell-form transition which depended on the NaCl concentration in the medium. In response to 0.8% NaCl addition into the medium, the pair/tetrad (designated 2/4) cells in a young culture grew and divided but did not separate and became 8-, 16-, and 32-cell units successively. In exponential growth phase, the cells divided in a 16/32 pattern. Potassium ions were equally effective as Na+ in mediating this multicell-formation effect; Mg2+, Li+, and Ca2+ also worked but produced less multiplicity. This effect appears to be species specific. This-section micrographs revealed that in a 16/32-cell unit, eight 2/4 cells were encased in an orderly manner within a large peripheral wall, showing five cycles of septation. Our results suggest the presence of a salt-sensitive mechanism for controlling cell separation in D. radiodurans.

Manganese(II) induces cell division and increases in superoxide dismutase and catalase activities in an aging deinococcal culture.

Addition of Mn(II) at 2.5 microM or higher to stationary-phase cultures of Deinococcus radiodurans IR was found to trigger at least three rounds of cell division. This Mn(II)-induced cell division (Mn-CD) did not occur when the culture was in the exponential or death phase. The Mn-CD effect produced daughter cells proportionally reduced in size, pigmentation, and radioresistance but proportionally increased in activity and amount of the oxygen toxicity defense enzymes superoxide dismutase and catalase. In addition, the concentration of an Mn-CD-induced protein was found to remain high throughout the entire Mn-CD phase. It was also found that an untreated culture exhibited a growth curve characterized by a very rapid exponential-stationary transition and that cells which had just reached the early stationary phase were synchronous. Our results suggest the presence of an Mn(II)-sensitive mechanism for controlling cell division. The Mn-CD effect appears to be specific to the cation Mn(II) and the radioresistant bacteria, deinococci.