The effects of microgravity on induced mutation in Escherichia coli and Saccharomyces cerevisiae.

We examined whether microgravity influences the induced-mutation frequencies through in vivo experiments during space flight aboard the space shuttle Discovery (STS-91). We prepared dried samples of repair-deficient strains and parental strains of Escherichia (E.) coli and Saccharomyces (S.) cerevisiae given DNA damage treatment. After culture in space, we measured the induced-mutation frequencies and SOS-responses under microgravity. The experimental findings indicate that almost the same induced-mutation frequencies and SOS-responses of space samples were observed in both strains compared with the ground control samples. It is suggested that microgravity might not influence induced-mutation frequencies and SOS-responses at the stages of DNA replication and/or DNA repair. In addition, we developed a new experimental apparatus for space experiments to culture and freeze stocks of E. coli and S. cerevisiae cells.     

Biodosimetry of heavyions by interphase chromosome painting

We report measurements of chromosomal aberrations in peripheral blood lymphocytes from cancer patients undergoingradiotherapy treatment. Patients with cervix or esophageal cancer were treated with 10 MV X-rays produced at a LINAC accelerator, or high-energy carbon ions produced at the HIMAC accelerator at the National Institute for Radiological Sciences (NIRS) in Chiba. Blood samples were obtained before, during, and after the radiation treatment. Chromosomes were prematurely condensed by incubation in calyculin A. Aberrations in chromosomes 2 and 4 were scored after fluorescence in situ hybridization with whole-chromosome probes. Pre-treatment samples were exposed in vitro to X-rays, individual dose-response curves for the induction of chromosomal aberrations were determined, and used as calibration curves to calculate the effective whole-body dose absorbed during the treatment. This calculated dose, based on the calibration curve relative to the induction of reciprocal exchanges, has a sharp increase after the first few fractions of the treatment, then saturates at high doses. Although carbon ions are 2–3 times more effective than X-rays in tumor sterilization, the effective dose was similar to that of X-ray treatment. However, the frequency of complex-type chromosomal exchanges was much higher for patients treated with carbon ions than X-ray.     

Countermeasure Against Regenerative and Forced Chatter of Flexible Workpieces in Milling Processs Using Bi-directional Excitation

A novel extended methodology for chatter suppression in milling process by applying external forced vibrations to the workpiece in two orthogonal directions which are the feed and cross-feed directions.Both the regenerative and forced chatter suppression during the milling process of flexible workpieces are investigated.Here,the workpiece is subject to a sinusoidal periodic force in the feed direction to disrupt the regenerative effect.Additionally,to minimize the forced chatter,the workpiece is subject to the periodic excitation force in cross-feed direction.This force is proportional to the magnitude of the estimated cutting force in cross-feed direction and has a phase opposite to the cutting force to minimize the vibration amplitudes.The effectiveness of the proposed method is evaluated numerically and experimentally,for the spindle speed located in both the local minima and local maxima of the stability lobe diagram.The numerical simulations indicate significant suppression effect in terms of vibration amplitudes,resulting in suppression of both the regenerative chatter and the forced chatter.Experiments were conducted by using a workpiece-mounted active stage composed of flexure hinges and driven by piezoelectric actuators.The experimental results agree qualitatively with the numerical simulations.The proposed method indicates a remarkable vibration reduction effect for both regenerative and forced chatters.     

Residual chromatin breaks as biodosimetry for cell killing by carbon ions

We have studied the relationship between cell killing and the induction of residual chromatin breaks on various human cell lines and primary cultured cells obtained by biopsy from patients irradiated with either X-rays or heavy-ion beams to identify potential bio-marker of radiosensitivity for radiation-induced cell killing. The carbon-ion beams were accelerated with the Heavy Ion Medical Accelerator in Chiba (HIMAC). Six primary cultures obtained by biopsy from 6 patients with carcinoma of the cervix were irradiated with two different mono-LET beams (LET= 13 keV/μm, 76 keV/μm) and 200kV X rays. Residual chromatin breaks were measured by counting the number of non-rejoining chromatin fragments detected by the premature chromosome condensation (PCC) technique after a 24 hour postirradiation incubation period. The induction rate of residual chromatin breaks per cell per Gy was the highest for 76 keV/μm beams on all of the cells. Our results indicated that cell which was more sensitive to the cell killing was similarly more susceptible to induction of residual chromatin breaks. Furthermore there is a good correlation between these two end points in various cell lines and primary cultured cells. This suggests that the detection of residual chromatin breaks by the PCC technique may be useful as a predictive assay of tumor response to cancer radiotherapy.     

DNA-lesion and cell death by alpha-particles and nitrogen ions.

When the natural logarithm of the surviving fraction is plotted against the dose of radiation, curves with shoulders at relatively high survival levels are obtained after gamma-rays. The curves were practically linear in case of HMV-I and HA-1 cells irradiated by charged particle beams. These cells were derived from human malignant melanoma and Chinese hamster cells, respectively. The amount of DNA single strand breaks (ssb) by gamma-rays or nitrogen-ions (LET=530KeV/micrometers) in HMV-I cells increases linearly with increment in dose, when the ssb is detected using the alkaline elution technique. There is no close relationship between the dose-response curve of the ssb and the dose-survival curves after gamma-rays or N-ions. The amount of DNA double strand breaks (dsb) by gamma-rays increases quadratically with increment of dose, in both HMV-I cells and HA-1 cells, when the dsb is detected using the neutral elution technique. The survival fraction for HA-1 cells is slightly higher than that for HMV-I cells, at the same dose, and the amount of dsb for HA-1 cells is considerably greater than that for HMV-I cells. These results suggest that the radiosensitivities to gamma-rays in different cell lines do not correspond to the number of DNA strand breaks. The amount of both non-repairable ssb and dsb also increases quadratically with increment of dose for gamma-rays and almost linearly with increment of dose for N-ions and alpha-particles (LET=36keV/micrometers for HA-1 cells and LET=77keV/micrometers for HMV-I cells). The dose-response curves for non-repairable dsb in case of these radiations seemed to mirror image the dose-survival curves for these radiations, in both cell lines. The number of non-repairable DNA strand breaks in the two cell lines, at the same level of survival was much the same. These results show the close relationship between the induction of non-repairable DNA strand breaks and cell killing.     

A balloon-borne very long baseline interferometry experiment in the stratosphere: Systems design and developments

The balloon-borne very long baseline interferometry (VLBI) experiment is a technical feasibility study for performing radio interferometry in the stratosphere. The flight model has been developed. A balloon-borne VLBI station will be launched to establish interferometric fringes with ground-based VLBI stations distributed over the Japanese islands at an observing frequency of approximately 20?GHz as the first step. This paper describes the system design and development of a series of observing instruments and bus systems. In addition to the advantages of avoiding the atmospheric effects of absorption and fluctuation in high frequency radio observation, the mobility of a station can improve the sampling coverage (“uv-coverage”) by increasing the number of baselines by the number of ground-based counterparts for each observation day. This benefit cannot be obtained with conventional arrays that solely comprise ground-based stations. The balloon-borne VLBI can contribute to a future progress of research fields such as black holes by direct imaging.     

Differentiation of Dictyostelium discoideum vegetative cells into spores during Earth orbit in space.

We reported previously that emerged amoebae of Dictyostelium (D.) discoideum grew, aggregated and differentiated to fruiting bodies with normal morphology in space. Here, we investigated the effects of space radiation and/or microgravity on the number, viability, kinetics of germination, growth rate and mutation frequency of spores formed in space in a radiation-sensitive strain, gamma s13, and the parental strain, NC4. In gamma s13, there were hardly spores in the fruiting bodies formed in space. In NC4, we found a decrease in the number of spores, a delay in germination of the spores and delayed start of cell growth of the spores formed in space when compared to the ground control. However, the mutation frequency of the NC4 spores formed in space was similar to that of the ground control. We conclude that the depression of spore formation might be induced by microgravity and/or space radiation through the depression of some stage(s) of DNA repair during cell differentiation in the slime mold.     

Nonlinear Inertial Guidance Platform Control Systems Analysis

This paper investigates the stability analysis of nonlinear inertial guidance platform control systems. The principle technique used for this investigation is the direct method of Lyapunov. First, stability criteria are derived for third- and fourth-order systems with saturation type nonlinearity by using the technique developed by R. E. Kalman and Z. V. Rekasius. A method of finding the maximum loop gain g/h is then shown, and the results are compared with those obtained by linear analyses. Finally, the stability criteria are extended to the cases when a coulomb friction type nonlinearity is included.     

Alkylating agent (MNU)-induced mutation in space environment.

In recent years, some contradictory data about the effects of microgravity on radiation-induced biological responses in space experiments have been reported. We prepared a damaged template DNA produced with an alkylating agent (N-methyl-N-nitroso urea; MNU) to measure incorrect base-incorporation during DNA replication in microgravity. We examined whether mutation frequency is affected by microgravity during DNA replication for a DNA template damaged by an alkylating agent. Using an in vitro enzymatic reaction system, DNA synthesis by Taq polymerase or polymerase III was done during a US space shuttle mission (Discovery, STS-91). After the flight, DNA replication and mutation frequencies were measured. We found that there was almost no effect of microgravity on DNA replication and mutation frequency. It is suggested that microgravity might not affect at the stage of substrate incorporation in induced-mutation frequency.     

Estimation of orbital parameters of broken-up objects from in-situ debris measurements

Even sub-millimeter-size debris could cause a fatal damage on a spacecraft. Such tiny debris cannot be followed up or tracked from the ground. Therefore, Kyushu University has initiated IDEA the project for In-situ Debris Environmental Awareness, which conducts in-situ measurements of sub-millimeter-size debris. One of the objectives is to estimate the location of on-orbit satellite fragmentations from in-situ measurements. The previous studies revealed that it is important to find out the right nodal precession rate to estimate the orbital parameters of a broken-up object properly. Therefore, this study derives a constraint equation that applies to the nodal precession rate of the broken-up object. This study also establishes an effective procedure to estimate properly the orbital parameters of a broken-up object with the constraint equation.