High current DC negative ion source for cyclotron.

A filament driven multi-cusp negative ion source has been developed for proton cyclotrons in medical applications. In Cs-free operation, continuous H(-) beam of 10 mA and D(-) beam of 3.3 mA were obtained stably at an arc-discharge power of 3 kW and 2.4 kW, respectively. In Cs-seeded operation, H(-) beam current reached 22 mA at a lower arc power of 2.6 kW with less co-extracted electron current. The optimum gas flow rate, which gives the highest H(-) current, was 15 sccm in the Cs-free operation, while it decreased to 4 sccm in the Cs-seeded operation. The relationship between H(-) production and the design/operating parameters has been also investigated by a numerical study with KEIO-MARC code, which gives a reasonable explanation to the experimental results of the H(-) current dependence on the arc power.

Development of a high current H(-) ion source for cyclotrons.

A multi-cusp DC H(-) ion source has been designed and fabricated for medical applications of cyclotrons. Optimization of the ion source is in progress, such as the improvement of the filament configuration, magnetic filter strength, extraction electrode's shape, configuration of electron suppression magnets, and plasma electrode material. A small quantity of Cs has been introduced into the ion source to enhance the negative ion beam current. The ion source produced 16 mA of DC H(-) ion beam with the Cs-seeded operation at a low arc discharge power of 2.8 kW.

Evaluation of thermal neutron irradiation field using a cyclotron-based neutron source for alpha autoradiography.

It is important to measure the microdistribution of (10)B in a cell to predict the cell-killing effect of new boron compounds in the field of boron neutron capture therapy. Alpha autoradiography has generally been used to detect the microdistribution of (10)B in a cell. Although it has been performed using a reactor-based neutron source, the realization of an accelerator-based thermal neutron irradiation field is anticipated because of its easy installation at any location and stable operation. Therefore, we propose a method using a cyclotron-based epithermal neutron source in combination with a water phantom to produce a thermal neutron irradiation field for alpha autoradiography. This system can supply a uniform thermal neutron field with an intensity of 1.7×10(9) (cm(-2)s(-1)) and an area of 40mm in diameter. In this paper, we give an overview of our proposed system and describe a demonstration test using a mouse liver sample injected with 500mg/kg of boronophenyl-alanine.

A phantom experiment for the evaluation of whole body exposure during BNCT using cyclotron-based epithermal neutron source (C-BENS).

At Kyoto University Research Reactor Institute (KURRI), cyclotron-based epithermal neutron source was installed in December 2008, and the supplementary construction works have been performed. As of December 2010, the various irradiation characteristics important for BNCT were mostly evaluated. The whole body exposure during BNCT medical irradiation is one of the important characteristics. In this article, measurements of absorbed dose for thermal and fast neutrons and gamma-ray at ten positions corresponding to important organs are reported.

Experimental verification of beam characteristics for cyclotron-based epithermal neutron source (C-BENS).

A cyclotron-based epithermal neutron source has been developed for boron neutron capture therapy. This system consists of a cyclotron accelerator producing 1.1-mA proton beams with an energy of 30 MeV, a beam transport system coupled with a beryllium neutron production target, and a beam-shaping assembly (BSA) with a neutron collimator. In our previous work, the BSA was optimized to obtain sufficient epithermal neutron fluxes of ~10(9) cm(-2) s(-1) using a Monte Carlo simulation code. In order to validate the simulation results, irradiation tests using multi-foil activation at the surface of a gamma-ray shield located behind the collimator and water phantom experiments using a collimated epithermal neutron beam were performed. It was confirmed experimentally that the intensity of the epithermal neutrons was 1.2×10(9) cm(-2) s(-1).

Evaluation for activities of component of Cyclotron-Based Epithermal Neutron Source (C-BENS) and the surface of concrete wall in irradiation room.

The workers employed in BNCT must enter the irradiation room just after an irradiation under the condition of remaining activities. To reduce the radiation exposure for the workers, it is important to identify the origins of the activities. In this research, the activities induced on the concrete wall surface were evaluated using MCNP-5 and the measurement results of thermal neutron distribution. Furthermore, the radioisotopes produced in the moderator were identified with a High Purity Germanium detector. It was found that the activities of the wall were mainly caused by (46)Sc, (60)Co and (152)Eu, and that (24)Na and (56)Mn were mainly produced in the moderator.

Importance of gated CT acquisition for the quantitative improvement of the gated PET/CT in moving phantom.

OBJECTIVE: The aim of this study was to investigate the utility of gated PET/CT and CT attenuation correction (AC) for the quantitation of radioactivity. METHODS: An ellipse phantom containing six spheres, ranging from 10 to 37 mm in diameter, was filled with 36.7 kBq/mL of F-18. The respiratory motion was simulated by a motor-driven plastic platform to move the phantom with a displacement of 2 cm in the craniocaudal direction at a frequency of 15/min. With the phantom at rest, PET/CT data were acquired and used as a standard (nonmotion). With the phantom in motion, PET data were acquired in both the static and gated modes (sPET and gPET, respectively). Helical CT (HCT), slow CT (SCT), average CT (ACT), and four-dimensional CT (4DCT) were acquired and used to correct attenuation. On both PET and CT images, the maximum radioactivity, dimensions, and CT numbers were measured on the central slices. RESULTS: In nonmotion, recovery coefficients whose spheres were 22 mm or smaller gradually decreased. Regarding motion, the PET counts of the spheres in the static acquisition were lower than those acquired in nonmotion with either type of CTAC (sPET-HCT: -43.8%, sPET-SCT: -51.4%, sPET-ACT: -49.5%). Gated acquisition of PET significantly improved the PET counts (gPET-HCT: -30.1%) (p < 0.05), while additional gated acquisition of CT significantly improved them further (gPET-4DCT: -15.2%) (p < 0.01). The dimensions of sPET were overestimated, but those of gPET were close to the standard values. The SCT significantly overestimated the dimensions, and the water density area decreased (p < 0.01). The 4DCT images were similar to the HCT images. CONCLUSIONS: In respiratory motion, PET acquisition in the static mode underestimated the radioactivity and overestimated the dimensions. Neither SCT nor ACT improved these errors. Although PET acquisition in the gated mode improved the quantification of PET/CT images, the additional gated CT acquisition using 4DCT is required for further improvement.

Improvement of dose distribution in phantom by using epithermal neutron source based on the Be(p,n) reaction using a 30 MeV proton cyclotron accelerator.

In order to generate epithermal neutrons for boron neutron capture therapy (BNCT), we proposed the method of filtering and moderating fast neutrons, which are emitted from the reaction between a beryllium target and 30 MeV protons accelerated by a cyclotron, using an optimum moderator system composed of iron, lead, aluminum, calcium fluoride, and enriched (6)LiF ceramic filter. At present, the epithermal-neutron source is under construction since June 2008 at Kyoto University Research Reactor Institute. This system consists of a cyclotron to supply a proton beam of about 1 mA at 30 MeV, a beam transport system, a beam scanner system for heat reduction on the beryllium target, a target cooling system, a beam shaping assembly, and an irradiation bed for patients. In this article, an overview of the cyclotron-based neutron source (CBNS) and the properties of the treatment neutron beam optimized by using the MCNPX Monte Carlo code are presented. The distribution of the RBE (relative biological effectiveness) dose in a phantom shows that, assuming a (10)B concentration of 13 ppm for normal tissue, this beam could be employed to treat a patient with an irradiation time less than 30 min and a dose less than 12.5 Gy-eq to normal tissue. The CBNS might be an alternative to the reactor-based neutron sources for BNCT treatments.

Combined effect of extracellular matrices and growth factors on bovine corneal endothelial cells in culture.

PURPOSE: First, to confirm that corneal endothelial cells in the confluent state have the capability to form cellular covering. Second, to establish a method to study the combined effect of extracellular matrices (ECMs) and growth factors on the biological response in corneal endothelial cells in culture. METHODS: Bovine corneal endothelial cells were cultured inside a cylinder set on a plastic dish. They formed a confluent cell nest on the dish coated with type I or type IV collagen, laminin, or fibronectin. After the removal of the cylinder, hepatocyte growth factor (HGF), epidermal growth factor, transforming growth factor-alpha or transforming growth factor-beta(1) was added to the cultures. Each confluent cell nest enlarged outward, and its increased area size was measured. Cellular response in the nest, including cellular proliferation, was analyzed. RESULTS: The size of the increased area of the culture on type IV collagen plus HGF was the largest of all the combinations of ECMs and growth factors. The responses of component cells in the increased area consisted of cellular hypertrophy, proliferation, migration and giant cell formation. The treatment with type IV collagen plus HGF clearly promoted all the above responses. CONCLUSIONS: The biological response of corneal endothelial cells was regulated by ECMs and growth factors.

Production and characterization of a bacterial single-chain Fv fragment specific to human truncated midkine.

The production (and characterization) of a monoclonal antibody against human truncated midkine (tMK), and the detection of tMK in G401 cells, a Wilms' tumor cell line, as well as in Wilms' tumor patient specimens, have been reported (Paul et al., Cancer Lett. 163 (2001) 245-251). Here we report the molecular cloning and expression of this monoclonal antibody as a single-chain Fv fragment (scFv) in Escherichia coli. The scFv protein, purified by immobilized metal affinity chromatography, showed a specific affinity to recombinant tMK and native tMK in G401 cells as detected by enzyme-linked immunosorbent assay and immunofluorescence microscopy, respectively. The binding of this protein to recombinant tMK was competitive with the parental monoclonal antibody. These results suggest that this scFv can also be used for Wilms' tumor detection.

Molecular cloning, expression and purification of truncated midkine and its growth stimulatory activity on Wilms' tumor (G401) cells.

Midkine (MK) is a heparin binding growth factor identified as a product of a retinoic acid-responsive gene; it is frequently expressed at high levels in many human carcinomas. Although the expression of the mRNA encoding truncated MK (tMK) in unique human cancer cells has been reported, the tMK polypeptide itself has not yet been identified. In order to clarify the biological role of tMK, recombinant tMK was expressed in Escherichia coli and purified. Recombinant tMK was purified as a single band in SDS-PAGE under reducing conditions showing an apparent molecular mass of 10 kDa. Purified recombinant tMK showed the same extent of proliferative activity towards Wilms' tumor (G401) cells as full length human MK. These results suggest that the structure of this recombinant tMK is same as the native polypeptide.

Detection of truncated midkine in Wilms' tumor by a monoclonal antibody against human recombinant truncated midkine.

Although the expression of a truncated midkine (tMK) mRNA has been detected in many cancer cells, the tMK protein itself has not yet been identified. The expression, purification and characterization of human recombinant tMK were described in the former report. A mouse hybridoma cell line producing an IgG2b monoclonal antibody (mab) against purified recombinant tMK was established. This anti-tMK mab did not cross react with synthetic full length (or c-half) human midkine. A putative native tMK was identified in G401 cells using this mab, and showed the same apparent Mw as the recombinant tMK in SDS-PAGE. This mab was also used in an immunohistochemical study to evaluate the expression of tMK in Wilms' tumor cell line, G401 cells, as well as in Wilms' tumor patient specimens. G401 cells and all Wilms' tumor patient specimens immunoreacted with this anti-tMK mab. We conclude that Wilms' tumor cells express tMK and that this mab is useful for the detection of tMK in the Wilms' tumor.

A factor that prevents EDTA-induced cell-growth inhibition: purification of transthyretin from chick embryo brain.

We previously reported the inhibition of cell-growth in Neuro-2A cells, mouse neuroblastoma, by Zn2+ chelation with EDTA. This paper describes the purification of a factor that prevents EDTA-induced cell-growth inhibition from chick embryo brain. The purified factor has a molecular mass of 16 kDa on SDS-polyacrylamide gel electrophoresis under reducing conditions. This factor prevents the cell-growth inhibition in a dose-dependent manner and also binds thyroxine. Analysis of the N-terminal amino acid sequence revealed that 40 residues coincide with the sequence of chicken liver transthyretin.

Effects of hepatocyte growth factor, transforming growth factor-beta1 and epidermal growth factor on bovine corneal epithelial cells under epithelial-keratocyte interaction in reconstruction culture.

In the cornea, corneal epithelial cells are in close contact with keratocytes: the epithelial cells organize thickened lamellar structure on a layer of keratocytes embedded in extracellular matrix (ECM). Thus, growth factors are expected to critically regulate corneal component cells under epithelial-keratocyte interaction. The purpose of this study is to clarify effects of hepatocyte growth factor (HGF), transforming growth factor-beta1 (TGF-beta1) or epidermal growth factor (EGF) on corneal epithelial cells under epithelial-keratocyte interaction. We examined proliferation and differentiation of the epithelial cells in a simple corneal reconstruction culture composed of an epithelial cell layer on the keratocyte-containing stromal layer, using three-dimensional collagen gel matrix culture. We observed the morphological change by phase contrast microscopy, and conducted histological and immunohistochemical examinations. The epithelial proliferation was examined by nuclear bromodeoxy-uridine (BrdU) uptake. In the reconstructed cornea under epithelial-keratocyte interaction, EGF-, TGF-beta1- and HGF-treated cells formed a thickened epithelial layer consisting of 5-6, 5-6 and 3-4 cells, respectively. In fact, both EGF and TGF-beta1 induced significantly higher intakes of nuclear BrdU of the epithelial cells than HGF. In lamellar differentiation of the epithelial cells, TGF-beta1- or HGF-treated cells formed a triple lamellar structure specific for the in vivo corneal epithelium: basal, middle and superficial layers are composed of cuboidal basal-like cells, spindle wing-like cells and flat superficial-like cells, respectively. TGF-beta1-treated cells formed a more markedly thickened epithelial layer than HGF-treated cells. In contrast, EGF formed a single lamellar structure consisting of cuboidal cells. These results suggest that those growth factors regulate proliferation and/or lamellar differentiation of corneal epithelial cells under epithelial-keratocyte interaction. The most interesting result was that TGF-beta1 promotes proliferation and lamellar differentiation of corneal epithelial cells through keratocyte-mediated stimulation.

Calcium-dependent potentiation of M-current in bullfrog sympathetic neurons.

Whole-cell voltage-clamp recordings were made from cultured bullfrog sympathetic neurons to measure the steady-state activation curve of M-type potassium current. When measured with a calcium-deficient (10 nM) pipette solution M-conductance was 4.8 nS at -35 mV having the 50%-activation voltage at-20 mV. Respective values were 17.2 nS at -35 mV with the 50%-activation voltage at -42 mV when measured with a calcium-rich (1 microM) solution, indicating the hyperpolarizing displacement of the activation curve with high internal calcium. It is suggested that intracellular calcium ions can modulate kinetics of M-current which thereby regulate the number of M-channels being open at given membrane potentials.

[Ovarian metastasis in patients with cervical cancer].

The study included a total of 634 patients with cervical cancer; 311 were in stage I b and 323 were in stage II. All patients received radical hysterectomy at the Department of Obstetrics and Gynecology, Kinki University, between May 1975 and December 1986. One out of 311 patients in stage I b (0.3%) and eight out of 323 patients in stage II (2.5%) had ovarian metastases confirmed histologically. Nine patients who had ovarian metastases were investigated about stages, histological types of the cervical cancer (WHO and CPL classification), incidence of metastases in pelvic lymph nodes, corpus infiltration, parametrial infiltration, and peritoneal cytology. Results obtained were as follows: 1. As to histological types, patients with adenocarcinoma and/or PL type in CPL classification had high incidence of ovarian metastases. 2. Patients with corpus infiltration had high incidence of ovarian metastases. 3. Metastases to both ovary and pelvic lymph nodes tended to occur on the same side, but ovarian metastases and parametrial infiltration did not necessarily occur on the same side. We suggest removal of ovaries, even before menopause, in patients with cervical cancer who have cervical adenocarcinoma, or corpus infiltration, or are suspicious for metastases in more than two pelvic lymph nodes.

Importance of albumen content in whole-body protein synthesis of the chicken embryo during incubation.

The importance of egg albumen content in whole-body protein synthesis was investigated in developing chicken embryos by using lines genetically selected for high and low albumen contents and by removing albumen from eggs before incubation. 2. Whole-body protein synthesis was estimated by injecting L-[15N]-phenylalanine intravenously on day 12 of incubation. 3. Embryos from high albumen eggs had higher whole-body protein synthesis rates than those from low albumen eggs. 4. Whole-body protein synthesis was reduced by the removal of albumen from eggs before incubation. 5. It was concluded that albumen content per se was of crucial importance in regulating whole-body protein synthesis in chicken embryos during incubation.

Significance of para-aortic node irradiation in the treatment of cervical cancer.

In 1980, we introduced para-aortic node irradiation as a part of our treatment of cervical cancer. Eighty-six patients with cervical cancer were selected for such treatment, based on histologic or radiologic criteria for lymph node involvement. We delivered 45 Gy to the para-aortic nodes in 5 weeks. Comparison studies on the prognosis and on the incidence of side effects with and without para-aortic node irradiation were carried out on a total of 783 patients who had invasive cervical cancer and were treated from 1975 to 1984. For patients in stages Ib and II who met, or before 1980 would have met, our criteria there was no significant difference in survival for those in stage Ib, but in stage II the para-aortic node irradiation group did significantly better (P less than 0.01). For the entire series of patients, for all stages from Ib to IV, the prognosis for the group treated after the introduction of para-aortic node irradiation was superior, with a statistically significant difference from the prognosis for patients treated before the introduction of this mode (Ib, P less than 0.05; II, P less than 0.01; III, P less than 0.01; IV, P less than 0.05). None of the patients had severe complications after para-aortic node irradiation. We conclude that, by irradiating the para-aortic nodes, we can improve the prognosis of cervical cancer patients who may have para-aortic node involvement, a situation which formerly was considered hopeless.