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We measured intestinal permeability of mice by PIXE method. A mixture of a rubidium chloride solution and a manganese chloride solution was administered orally into mice. The blood was sampled successively after the administration, and the element contents in the blood were measured by PIXE method. The Rb content in the plasma of Rb + Mn administered group reaches maximum 5 min after the administration and decreases subsequently. The value in the whole blood of Rb + Mn administered group increases slightly for first 30 min after the administration and remarkably from 3 hrs. This increase continues for 48 hrs. That in control group is very little. Rb is probably transferred into plasma from intestinal mucosa at first, and subsequently moves into blood cells. The Mn content in control group is little. The values in the whole blood and the plasma of Rb + Mn administered group reach maximum 5 min after the administration and then decrease, and turn to decline to control level at 24 hrs. Mn is probably transferred into blood from intestinal mucosa throughout, and washed out more rapidly than Rb . The element contents of Rb and Mn in the plasma of Rb + Mn administered mice are found to increase to the peak level 5 min after the administration. Rb and Mn are useful as a tracer to study of intestinal disease with measurement by PIXE method. With this manner by PIXE method, intestinal permeability can be measured more briefly than existing methods. In order to assess intestinal permeability, we propose an indicator which is obtained by comparing the element amounts of Rb and/or Mn in blood sampled just after the administration.

The quantitative changes of elements in blood plasma were observed with the passage of time after X-ray whole body irradiation with 12 Gy on C57BL/6J mice by PIXE method. From 4 days after irradiation, dead mouse was found and all mice died by 8 days. Hematocrit (Ht) values indicated a decrease from the 1st day, but on days 3 and 4 there was a small rise. Finally the values became 64 % of that of non-irradiated control on day 7, it was just before death. By analysis with PIXE method, 15 elements were observable in blood plasma of control mice. The elements such as P, S, Cl, K, Ca and Cr were abundant and Fe and Br followed. As trace elements, the peaks of Zn, Cu and Ni were clearly observed. After irradiation, K and Ca decreased on day 1st, afterwards increased gradually. On the contrary, the elements, S, Cl, were rather stable. While Fe decreased from 1st day, Cu increased from the day 2. Zn and Ni showed intensely down and rise in amount, and decreased on day 7. The results of possible measurement of the changes in amount of these elements of blood plasma suggest PIXE method is an easy and useful way for diagnosis.

The quantitative changes in the elements, amounts of Cl, K, Ca, in blood plasma were measured by PIXE method. The samples were obtained at appropriate intervals after transplantation of EL-4 tumor cells in three strains of mice, C57BL/6J (H-2b), C57BL/10J (abbreviation: B10; H-2b) and A/J (H-2a). Transplanted EL-4 tumor cells proliferated in both strains of C57BL/6J and B10. In A/J mice, transplanted EL-4 cells proliferated about 10 days and then were rejected completely by the immunological reaction according to the difference of major histocompatibility antigens. The amounts of Cl in plasma remained at similar level in the time course in any strains, but K fluctuated in C57BL/6J and B10, and less in A/J. On the other hand, Ca showed always higher values in C57BL/6J than other two strains of mice. In B10 mice, Ca increased just before death, but in A/J it decreased at the time of healing by rejection. These changes of Ca in the three strains of mice were related quantitatively 10 the hematocrit values of these strains of mice after transplantation of EL-4 cells.

Constitutional elements of head hair of mouse (Mus musculus) were investigated by Particle-Induced X-ray Emission (PIXE) analysis. The high peaks of S, K and Ca, and the low peaks of several other elements were observed in PIXE spectra of all the specimens from the used strains. The result clearly indicates that the ratios of these elemental contents are different among the coat colors of mice. Further, we observed strain difference even in the same color.

We investigated the elemental composition of mouse blood plasma with PIXE method. We measured quantitative X-ray yields of S, Cl, K and Ca by changing sample volume from 1.25 to 15.0 µl to find the most profitable volume of plasma for elemental analysis. Linear relationships were confirmed between the applied volume (1.25~5.0 µl) and the counts of X-ray from each element. Some elements showed, however, saturated curves over 5.0 µl. Five μl was enough to analyze some important elements of blood plasma and used in the present analysis. We propose 5.0 µl as a standard amount of sample in plasma analysis. We found differences in the elemental ratio of X-ray counts in plasma among five mouse strains used in the present experiment. This result suggests that the strain differences in the elemental constitution of mouse blood plasma are related to those of physiological states such as the number of blood cells and the radio sensitivity.

PIXE application to the measurement of cellular elements is outlined on characteristic variations of the contents of yeast, CHO, V79 and MM46 cultured mammalian cells according to environmental changes. Cellular elements from P to Br were successfully analyzed at these cells after cautious preparation of samples in filtration steps with as possible as less deterioration. We confirmed linear relationships between the X-ray yields and these element contents. The experiment was extended to an analysis of cellular substances at molecular level by scanning of specimens. Preliminary results were included.

PIXE analysis was successfully applied to the measurement of cellular elements of mammalian cells in relation to the study of hyperthermia. V79 cells were heated at 44.0°C for 30 min or 1 h and the following element releases or uptakes were determined with PIXE measurement of samples obtained after appropriate periods of incubation at 37°C. Linear relationships were confirmed between the produced X-ray yields of P, S, Cl, K and Ca and the cell numbers. The cellular contents P, S, and K decreased promptly after the heat treatment and recovered gradually in the following incubation. Changes in the contents of Cl and Ca did not show abrupt decreases. Curiously, the content of Ca increased after the treatment at 44.0°C for 30 min. The results suggest that various structural modifications were induced in the membrane by the heat treatment.