Investigation with Bromsulphalein test and determination of urobilinogen in the urine was carried out in men, who are living in Spa Kusatsu in good condition, and take thermal bath once or twice everyday.
Bromsulphalein excretion was enhanced by the stay in the Spa within 4 months and the amount of urobilinogen increased after 5 months, but there was no significant correlation between them.

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Serum iron was determined by Dipyridil method. Serum Copper was measured by Gubler's Diethyl-dithiocarbamate method.
(1) A significant diurnal variation was recognized in serum iron and copper of six adult subjects. The serum iron and copper showed the highest level at noon and the lowest at midnight. Namely average serum iron content was 122γ/100ml at 12. a. m. and 84γ/100ml at 12. p. m.
Average serum copper content was 102γ/100ml at 12. a. m. and 89γ/100ml at 12. p. m.
(2) Iron and copper in the blood serum were measured in seven healthy adults before and after a thermal bath in Ikaho Hot Spring and in plain water respectively at 42°C for 10 minutes. But no significant change was proved.
(3) Serum iron and copper in 9 healthy subjects were increased by acethylcholin injection. Serum iron rose 20 minutes after the injection (149γ/100ml), and returned to the former level after 60 minutes (131γ/100ml). Serum copper reached the highest value 30 minutes after the injection (111γ/100ml), and returned to the initial level after 60 minutes (100γ/ml).
(4) Serum iron and copper in 6 healthy subjects were decreased by adrenalin injection. Serum iron fell to 95γ/ml 30 minutes after the injection, and returned to the former level after 60 minutes (05γ/100ml). Serum copper fell to the lowest level 30 minutes after the injection (80γ/100ml) and returned to the former level after 60 minutes (101γ/100ml).
(5) Chlorpromazine injection decreased the serum iron in 5 subjects after 30 minutes, but it recovered to the initial level in 60 minutes.
No significant change in serum copper was caused by chlorpromazine injection. Chlorpromazine seems to suppress the diurnal variation in serum iron and copper.

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Using radioactive iron(Fe⁵⁹), the author studied the effects of the changes in bath conditions on the percutaneous absorption of iron in the bath water.
Radioactive iron was dissolved in a solution of hydrochloric acid, as ferric chloride. One tenth ml of this solution contained about 10 microcuries of Fe⁵⁹. From 0.5ml to 1.0ml of this solution was added to 150ml of bath water.
Mice were bathed in the bath water thus labeled with Fe⁵⁹.
The animal was fixed by tying to a wire-netting during the bath so as not to drink bath water. As soon as the bath was over, the test-animals were killed by strangulation. Then in order to remove contaminated Fe⁵⁹ on the skin, mice were washed three times with dilute hydrochloride solution and then three times by water. Skinned mice were ignited to ashes completely in porcelain crucible in 5 to 7 hours at 500°C. The ash was dissolved in 3 normal hydrochloric acid solution, transfered into a stainless plate, neutralized by ammoniac solution, methylred as an indicator. Then the precipitate was dried gradually.
Radioactivity of the precipitate was measured with Lauritsen's electroscope and following results were obtained.
1) Percutaneous absorption of ferric ions was stronger at a water temperature of 42°C than at 37°C.
2) Percutaneous absorption of iron into the body proved strongest with alkaline reaction (PH 10.0), weakest with acid reaction (PH 2.9) and moderate with weakly acid reaction (PH 6.5).
3) A linear correlation was roughly established between the duration of bath and the amount of the percutaneously absorbed iron.
4) No definite influence on the rate of percutaneous absorption of iron was proved by the repetition of thermal bath in Ikaho Hot Spring (a calcium sulphate water containing minute amount of iron).
5) Accumulation of iron in the body was proved by the serial bathings for 2-10 days in the ferric ion solution labeled with Fe⁵⁹.
6) The percutaneous absorption of iron was not increased by the addition of reducing substances such as ascorbic acid or hydroxylamines into the bath water containing ferric ions.

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