Utility of a skin marker-less setup procedure using surface-guided imaging: a comparison with the traditional laser-based setup in extremity irradiation.

This study aimed to assess the feasibility of a skin marker-less patient setup using a surface-guided radiotherapy (SGRT) system for extremity radiotherapy. Twenty-five patients who underwent radiotherapy to the extremities were included in this retrospective study. The first group consisted of 10 patients and underwent a traditional setup procedure using skin marks and lasers. The second group comprised 15 patients and had a skin marker-less setup procedure that used an SGRT system only. To compare the two setup procedures for setup accuracy, the mean 3D vector shift magnitude was 0.9 mm for the traditional setup procedure and 0.5 mm for the skin marker-less setup procedure (p < 0.01). In addition, SGRT systems have been suggested to improve the accuracy and reproducibility of patient setups and consistently reduce interfractional setup errors. These results indicate that a skin marker-less patient setup procedure using an SGRT system is useful for extremity irradiation.

Tomography of the 2016 Kumamoto earthquake area and the Beppu-Shimabara graben.

Detailed three-dimensional images of P and S wave velocity and Poisson's ratio (σ) of the crust and upper mantle beneath Kyushu in SW Japan are determined, with a focus on the source area of the 2016 Kumamoto earthquake (M 7.3) that occurred in the Beppu-Shimabara graben (BSG) where four active volcanoes and many active faults exist. The 2016 Kumamoto earthquake took place in a high-velocity and low-σ zone in the upper crust, which is surrounded and underlain by low-velocity and high-σ anomalies in the upper mantle. This result suggests that, in and around the source zone of the 2016 Kumamoto earthquake, strong structural heterogeneities relating to active volcanoes and magmatic fluids exist, which may affect the seismogenesis. Along the BSG, low-velocity and high-σ anomalies do not exist everywhere in the upper mantle but mainly beneath the active volcanoes, suggesting that hot mantle upwelling is not the only cause of the graben. The BSG was most likely formed by joint effects of northward extension of the Okinawa Trough, westward extension of the Median Tectonic Line, and hot upwelling flow in the mantle wedge beneath the active volcanoes.

Development of a real-time imaging system for hypoxic cell apoptosis.

Hypoxic regions within the tumor form due to imbalances between cell proliferation and angiogenesis; specifically, temporary closure or a reduced flow due to abnormal vasculature. They create environments where cancer cells acquire resistance to therapies. Therefore, the development of therapeutic approaches targeting the hypoxic cells is one of the most crucial challenges for cancer regression. Screening potential candidates for effective diagnostic modalities even under a hypoxic environment would be an important first step. In this study, we describe the development of a real-time imaging system to monitor hypoxic cell apoptosis for such screening. The imaging system is composed of a cyclic luciferase (luc) gene under the control of an improved hypoxic-responsive promoter. The cyclic luc gene product works as a caspase-3 (cas-3) monitor as it gains luc activity in response to cas-3 activation. The promoter composed of six hypoxic responsible elements and the CMV IE1 core promoter drives the effective expression of the cyclic luc gene in hypoxic conditions, enhancing hypoxic cell apoptosis visualization. We also confirmed real-time imaging of hypoxic cell apoptosis in the spheroid, which shares properties with the tumor. Thus, this constructed system could be a powerful tool for the development of effective anticancer diagnostic modalities.

Determination of the relative biological effectiveness and oxygen enhancement ratio for micronuclei formation using high-LET radiation in solid tumor cells: An in vitro and in vivo study.

We determined the relative biological effectiveness (RBE) and oxygen enhancement ratio (OER) of micronuclei (MN) formation in clamped (hypoxic) and non-clamped (normoxic) solid tumors in mice legs following exposure to X-rays and heavy ions. Single-cell suspensions (aerobic) of non-irradiated tumors were prepared in parallel and used directly to determine the radiation response for aerobic cells. Squamous cell carcinoma (SCCVII) cells were transplanted into the right hind legs of syngeneic C3H/He male mice. Irradiation doses with either X-rays or heavy ions at a dose-averaged LET (linear energy transfer) of 14-192keV/µm were delivered to 5-mm diameter tumors and aerobic single-cells in sample-tubes. After irradiation, the tumors were excised and trypsinized to observe MN in single-cells. The single-cell suspensions were used for MN formation assays. The RBE values increased with increasing LET. The maximum RBE values for the three different oxygen conditions; hypoxic tumor, normoxic tumor, and aerobic cells, were 8.18, 5.30, and 3.76 at an LET of 192keV/µm, respectively. After X-irradiation, the OERh/n values (hypoxic tumor/normoxic tumor) were lower than the OERh/a (hypoxic tumor/aerobic cells), and were 1.73 and 2.58, respectively. We found that the OER for the in vivo studies were smaller in comparison to that for the in vitro studies. Both of the OER values at 192keV/µm were small in comparison to those of the X-ray irradiated samples. The OERh/n and OERh/a values at 192keV/µm were 1.12 and 1.19, respectively. Our results suggest that high LET radiation has a large biological effect even if a solid tumor includes substantial numbers of hypoxic cells. To conclude, we found that the RBE values under each oxygen state for non-MN fraction increased with increasing LET and that the OER values for both tumors in vivo and cells in vitro decreased with increasing LET.

Selective enhancement of hypoxic cell killing by tempol-regulated suicide gene expression.

The presence of hypoxic regions within solid tumors is caused by an imbalance between cell proliferation and angiogenesis. Such regions may facilitate the onset of recurrence after radiation therapy and chemotherapy, as hypoxic cells show resistance to these treatments. We found that tempol, a nitroxide, strongly induces the accumulation of hypoxia-inducible factor (HIF)-1α, particularly under conditions of hypoxia. We, therefore, evaluated whether tempol enhances the gene expression via HIF-1α, potentially leading to various applications for cancer gene therapy targeting hypoxic cells. Consequently, following treatment with tempol under hypoxia, the luciferase (Luc) activity in the cells transfected with the plasmid containing the luc gene with the oxygen-dependent degradation domain and a promoter composed of hypoxia-responsive elements increased up to approximately 10-fold compared to that observed in cells treated identically with the exception of tempol. The plasmid constructed by replacing the luc gene with the fcy::fur fusion gene as a suicide gene, strongly induced the accumulation of the Fcy::Fur fusion protein, only when incubated in the presence of the hypoxic mimic CoCl2 and tempol. The transfected cells were successfully killed with the addition of 5-fluorocytosine to the cell culture according to the fcy::fur fusion gene expression. As similar but lesser enhancement of the Luc activity was also observed in solid tumor tissues in nude mice, this strategy may be applied for hypoxic cancer eradication.

Evaluation of SCCVII tumor cell survival in clamped and non-clamped solid tumors exposed to carbon-ion beams in comparison to X-rays.

The aim of this study was to measure the RBE (relative biological effectiveness) and OER (oxygen enhancement ratio) for survival of cells within implanted solid tumors following exposure to 290MeV/nucleon carbon-ion beams or X-rays. Squamous cell carcinoma cells (SCCVII) were transplanted into the right hind legs of syngeneic C3H male mice. Irradiation with either carbon-ion beams with a 6-cm spread-out Bragg peak (SOBP, at 46 and 80keV/µm) or X-rays was delivered to 5-mm or less diameter tumors. We defined three different oxygen statuses of the irradiated cells. Hypoxic and normoxic conditions in tumors were produced by clamping or not clamping the leg to avoid blood flow. Furthermore, single-cell suspensions were prepared from non-irradiated tumors and directly used to determine the radiation response of aerobic cells. Single-cell suspensions (aerobic condition) were fully air-saturated. Single-cell suspensions were prepared from excised and trypsinized tumors, and were used for in vivo-in vitro colony formation assays to obtain cell survival curves. The RBE values increased with increasing LET in SOBP beams. The maximum RBE values in three different oxygen conditions; hypoxic tumor, normoxic tumor and aerobic cells, were 2.16, 1.76 and 1.66 at an LET of 80keV/µm, respectively. After X-ray irradiation the OERh/n values (hypoxic tumor/normoxic tumor) were lower than the OERh/a (hypoxic tumor/aerobic cells), and were 1.87±0.13 and 2.52±0.11, respectively. The OER values of carbon-ion irradiated samples were small in comparison to those of X-ray irradiated samples. However, no significant changes of the OER at proximal and distal positions within the SOBP carbon-ion beams were observed. To conclude, we found that the RBE values for cell survival increased with increasing LET and that the OER values changed little with increasing LET within the SOBP carbon-ion beams.

A luminescent single-molecule magnet: observation of magnetic anisotropy using emission as a probe.

An assemblage of a Tb(III) ion and two mononuclear [Zn(II)(L)] complexes as antenna chromophores (H(2)L: Schiff base ligand) afforded a luminescent single-molecule magnet. The f-f emission spectrum had partially resolved fine structure which provides detailed information on the magnetic anisotropy of the (7)F(6) ground multiplet of the Tb(III) ion.

Linear trinuclear Zn(II)-Ce(III)-Zn(II) complex which behaves as a single-molecule magnet.

Linear Zn(II)-Ce(III)-Zn(II) complex, which involves only one 4f electron as a spin source, behaves as an SMM. Easy-axis magnetic anisotropy for the ground (2)F(5/2) state of Ce(III) was achieved by a uni-axial crystal field, which is formed with four phenoxo oxygens as axial donors with the other five oxygens as equatorial donors.

Correlation between slow magnetic relaxation and the coordination structures of a family of linear trinuclear Zn(II)-Ln(III)-Zn(II) complexes (Ln = Tb, Dy, Ho, Er, Tm and Yb).

Six linear trinuclear [Ln{Zn(L)(AcO)}(2)]BPh(4) complexes (H(2)L denotes the Schiff-base ligand formed by a condensation reaction between ethylenediamine and two equivalents of o-vanillin), including Ln = Tb (1), Dy (2), Ho (3), Er (4), Tm (5) and Yb (6) were synthesized and were confirmed to be isostructural via X-ray crystallographic analyses. The Ln(III) ion in each complex is deca-coordinated by four equatorial oxygen donors from the methoxo groups of the Schiff-base ligands, two oxygen donors from the acetate anions and four axial oxygen donors from the phenoxo groups of the Schiff-base ligands. AC susceptibility measurements, with an oscillating frequency of 10 to 10,000 Hz, revealed that 1, 2, 4 and 6 show slow magnetic relaxation under a 1000 Oe DC bias field, which occurs via a single process, as confirmed by the semi-circular Cole-Cole plots. These complexes are considered to be field-induced single-molecule magnets under these conditions. The presence or absence of the slow magnetic relaxation process is discussed by correlating the characteristic magnetic anisotropy of each Ln(III) ion with the ligand field anisotropy.