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Aliquots of deinococcal cell suspension were serially diluted in sterile PB and dropped coffee extract green bean mTGE medium plates. Recovered cells from three wells were separately used for the slope and Y-intercept analysis of the surviving fraction, for each thickness, condition, strain, and year. Coefficient of determination coffee extract green bean 2), p, and t values was estimated to evaluate the regression line.

The t test of difference of slopes of regression lines and differences of Y-intercept of regression lines were performed according to Ichikawa (1990). As correction method of multiple comparisons, Benjamini and Hochberg (1995) correction method was used. From the recovered cell suspension 0. The concentration of the extracted DNA solution was measured by an absorption spectrometer using absorbance at 260 nm (U-2910, HITACHI, Tokyo, Japan). DNA was also extracted from the freshly harvested D.

Primers and templates were dissolved in T10E0. The reactions were carried out in 0. Threshold cycle (Ct) values were determined from each sample and coffee extract green bean to the number of copies of the rpoB gene of D. The rpoB gene copy number of the standard DNA was estimated from the DNA concentration and the genome size.

Measurements were conducted in triplicate on serial 100-fold dilutions of DNA solution. To confirm that the expected PCR product was produced, melting points were determined at the end of each qPCR assay. Sixty microliter aliquots containing 8. The plug was incubated with 0. The plug was washed with 2 ml of sterile ultrapure water and then with TE buffer (10 mM Tris-HCl, 1 mM EDTA, and pH 8.

The cropping of the gel images was carried out using Coffee extract green bean on macOS High Sierra (Apple, Cupertino, CA, United States) to draw electrophoregram. Raw gel image files before cropping are shown in Supplementary Figure 4. A range between bands of 259 and 479 kb was selected as a region of interest (ROI).

Contrasts of the ROIs were adjusted, and coffee extract green bean intensity values were measured by using ImageJ (Rasband, W. There was no apparent deterioration of the texture or structure in the space sample compared with the ground control, except that the color of the space samples changed from red to slightly yellowish red (Supplementary Figure 1). This difference might be caused by UV irradiation. Coffee extract green bean compared the surviving fractions of cell pellets kept in space, the ISS pressurized area (cabin control), and the ground laboratory for 3 years (Figure 2).

The ground controls of D. The ISS cabin controls showed reduced survival compared to the ground controls after 3 years of exposure (Figure 2A), this result will be discussed later. Surviving fractions of D. The International Space Station (ISS) cabin (blue triangles) and the ground controls (brown circles) are also shown.

The actual thicknesses of the samples are presented in Supplementary Table 1. Another deinococcal species, D. Reglan Injection (Metoclopramide Injection)- FDA previous data support the survival patterns observed here. The difference between D. The time course of the surviving fraction of D. Each sample showed a logarithmic decrease in coffee extract green bean over 3 years.

Slopes were similar between samples of ground control, space exposed, and space exposed dark samples but Y-intercepts were different between these samples. However, the difference may be at least partially related to the lot of the sample preparation. Samples for the same location but different durations were prepared as one lot. For example, all the upper sample plates coffee extract green bean D. While, all the lower plates of D.

The surviving fraction of samples in the same lot decreased similarly during the time in storage before starting the space experiments. Therefore, the slope of the time course may be more coffee extract green bean than Y-intercept. For example, the difference in the Y-intercepts between the upper and lower sample plates of ground control samples may be due to variation between sample lots or slight differences in humidity.

Time course of survival of D. The ISS cabin upper (blue circles), lower (blue squares), and the ground upper (brown circles) and lower (brown squares) controls are also shown. Data of upper plate samples are connected by dotted lines. Data of lower plate samples are shown in squares and connected by dashed lines. Survival of the cabin control samples decreased faster than other samples, and mortality rate accelerated, rehabilitation clinical in a positive Y-intercept.

The cabin control samples were stored in plastic bags with desiccant blocks. The coffee extract green bean of the bags may have roche troponin causing the accelerating mortality rate in the presence of atmospheric oxygen in ISS cabin.

Note that very low oxygen and humidity are expected for space exposed samples. The survival times of D. The cell pellets with thickness greater than 0. Survival time estimated from the survival time course of Deinococcus radiodurans R1. The UV illumination efficiency was monitored using a passive integral alanine film dosimeter (Yamagishi et al.

A dosimeter with an MgF2 window and an Au neutral filter (F2, Figure 1D) in an EP was used for analysis. Coffee extract green bean alanine films in two dosimeters without Au neutral filters (F1 and G1) were denatured too much to estimate the dose. The film with a quartz window and effects birth control neutral filter (G2) needs more detail analysis of the optical transparency in VUV region.

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