It has been generally recognized that balneotherapy is effective for anemia, especially drinking of ferruginous spring is quite effective for iron deficiency anemia. Although considerable morphological researches have been done in the field, the reports dealing with the biochemical analysis are few in number. Accordingly, the mechanism of the action of hot spring on hematopoietic function still remains obscured. For this reason, the author has made an attempt to observe the iron metabolism in order to clarify the mechanism.
Part I is concerned with critical studies in methods for iron determination.
1) Hemolysis easily occurs in the process of separation of rabbit serum, but usage of a centrifuge tube coated with “solid paraffin” helps to prevent this. Another advantage of this simple method is to increase the amount of serum obtained.
2) Compared with sensitivities of various kinds of chromogenic agents employed in iron determination, nitroso-R salt (α-Nitroso-β-naphthol-3, 6-disulfonic Acid Sodium Salt) was greater in sensitivity than any other, as already reported by other workers.
As for the factors which have influence upon the time required for the complete color development in nitroso-R salt, the wave-length and the addition of hydroxylamin have been already described, and the author pointed out temperature as the most important factor. From the author's experiments, it was confirmed that when temperature was kept at 37.0°C, complete color development occurred and kept its stability in 10 to 15 hours.
3) The serum iron value, obtained by Ramsay's method, was easily influenced by hemolysis. Accordingly, the author tried to find the relation between the measurement error of serum iron and the degree of hemolysis.
This attempt may be convenient in estimating the iron value in a hemolysed specimen.
4) Considering the above, some alterations were made on the routine optical method for the determination of unsaturated iron binding capacity (UIBC).
The mean value for UIBC of healthy mature rabbits was 117.3γ/dl.
5) The author devised a new micromethod for determination of ferrous and ferric ion in iron solution.
This method is also applicable to ferruginous spring in which permanganate titration is not employed owing to coexisting reducing materials.

Reports are scant concerning the influence of hot-spring bathing upon hematopoietic function.
The author attempted to observe the variation in serum iron by single bathing. Since rabbit serum iron value is changeable by the way of venesection itself and this could be connected with iron metabolism in a case of acute bleeding, the author studies this problem in the preliminary experiment. The results obtained are summarized as follows:
1) In animal experiments (rabbit), it is desirable to limit the number of blood collection to two, and the amount of initial sample to less than 4ml for the observation of serum iron changes to minimize influence of the removal of blood. And it is necessary to give careful consideration to the difference in serum iron values if determination was made after the long interval.
2) In acute bleeding, the decrease of serum iron at the beginning of bleeding was due simply to hydremia, associated with the loss of blood. In the subsequent course, however, serum iron value tended to be considerably complicated by the flow of storage iron into blood stream or the stasis in hematopoietic organs, but, on the whole, serum iron tended to decrease gradually.
Iron binding protein, at the beginning of bleeding, decreased in proportion to serum protein and the rise in the saturation per cent was temporarily resulted from the tendency to maintain constancy in serum iron. Subsequently, the saturation per cent gradually decreased.
And in the early stage such as 2 to 3 days after bleeding, iron binding protein remarkably increased before the recovery of serum protein, and the rate of increase was 25.8per cent.
From the viewpoint of iron absorption, this may be considered to be a vital reaction which serves the purpose.
3) Reports are few concerning the serum iron value of the rabbit, and the author measured the serum iron values in 118 healthy mature rabbits. The results are as follows:
_??_251.4±13.2γ/dl _??_211.5±14.9γ/dl (by Yoshikawa's method),
_??_205.9±14.8γ/dl _??_ 194.1±15.2γ/dl (by Matsubara's method),
_??_198.5±11.4γ/dl _??_199.4±10.9γ/dl (by Noda's method).
4) Single bathing in “Genzc-yu” (acid vitriol spring), “Unagi-yu” (alkaline sulphur spring) and simple carbondioxated spring at Narugo Spring showed no significant influence on the serum iron in healthy persons and rabbits.

Morphological studies have revealed the fact that drinking of ferruginous Water is as remarkably effective as large doses of reduced iron in iron deficiency anemia, in spite of extremely slight quantity of iron contained.
The author attempted biochemically to observe the intestinal iron absorption, which is the first step in the effect of drinking of ferruginous spring. The results obtained are summarized as follows:
1) It was found that the routine experiments with rabbits on intestinal iron absorption in which the rise of serum iron or tissue iron was measured as an index of absorption were not sufficiently dependable for observing the immediate absorption from the intestinal tract.
2) Serum iron has been described by other workers, as a factor involved in iron absorption.
From the author's experiments on anemic rabbit phlebotomized, it was suggested that iron absorption was influenced by the level of unsaturated iron-binding capacity, and there is a definite relationship between them.
3) The author devised what is called “supra-surviving double intestinal loop method”, an improved experimental wethal in intestinal iron absorption.
(1) Based on the withdrawal-test, it was presumed that iron entering into blood stream and iron accumulated in the jejunal mucosa were included in the quantity of absorption obtained by this method.
(2) There was considerable variations in the absorptive power of individual animals. On the other hand, the absorptive capacity of the upper jejunum in the same individual showed no significant difference, and the neighbouring two portions showed still less difference.
Strictly speaking, in the upper jejunum, the absorptive capacity of the lower area was slightly inferior to that of the upper.
It may be caused by the difference in the distribution of mesenteric blood vessel.
(3) The quantity of absorption was approximately proportional to the concentration of iron solution, injected into the intestinal loop (under 1500γ/10ml).
(4) When original “Genzo-yu” (pH 1.65) or H₂SO₄ solution (pH 1.60) was injected into the intestinal loop, pathologically serious changes developed in the jejunal mucosa. But these solutions, when diluted 4 or 5 times, showed no change.
(5) Absorption was influenced by pH of injected iron solution, and it was most remarkably accelerated in iron solution showing a pH of 1.15, which caused serious changes in the mucosa.
On the basis of further studies into the conflicting phenomena, it was evident that iron absorption in these experiments was considerably limited by secretion of intestinal juice.
4) By comparing the absorption of ferrugino s spring (“Genzo-yu”) water with that of FeSO₄ solution of the same concentration by this method, no significant difference was observed.
5) In this experimental method, some problems to be criticized was elicited and the author feels that it is probably unreasonable to make an inference from these experiments on iron absorption under physiological conditions.

The author studied the significance of the hot-spring bathing treatment upon rheumatic disease. Excreted urinary 17KS was measured for this purpose and Nomin-no-yu (sulfur spring) and thermal water of the University Hospital (sulfated spring), were employed. The results were as follows:
1) Compared with healthy persons, the value of excreted urinary 17KS of rheumatoid arthritis patients was low. Dividing rheumatoid arthritis patients into two groups, namely, those suffering from less than a year and those more than a year, it was found that no difference was observable in the values of urinary 17KS secretion between the former group and healthy persons and that the value was low in the latter group. Therefore it was postulated that the pituitary adrenocortical function might be impaired due to the prolonged process of rheumatic disease.
2) Hot-spring bathing treatment increased excreted urinary 17KS in rheumatoid arthritis patients. The increase in urinary 17KS excretion reached a maximum 7 days after the initiation of the treatment. After this it gradually decreased, though the increase was still observable 14 days after the initiation. It might be due to the difference in properties between Nomin-no-yu and thermal water of the University Hospital that the increase in excreted urinary 17KS was higher and the duration of the increase was longer in the former than the latter. It seems that the incerase was mainly due to vitalization of the pituitary adrenocortical function by nonspecific stimulation, because the difference was observed in individual reactions against synthetic stimulating action of hot-spring bathing.
3) The increase in excreted urinary 17KS caused by hot-spring bathing was less in rheumatoid arthritis patients than in healthy persons, In a very few cases of rheumatoid arthritis, hardly any increase could be observed; there were some cases in which a decrease in excreted urinary 17KS was observed. An inference can be made from the findings mentioned above that the pituitary adrenocortical function is generally deteriorated in rheumatoid arthritis patients.
4) Hot-spring bathing will increase excreted urinary 17KS in rheumatoid arthritis patients, if it is used in combination with aspirin therapy in which a large quantity of aspirin is used. Where this combined therapy was successful, a pronounced increase in excreted urinary 17KS was observed in rheumatoid arthritis patients. However, where the combined therapy was unsuccessful, the number of cases in which the increase was observed was almost equal to that in which the decrease was observed. These findings suggest that balneotherapy and aspirin therapy work mutually in the therapeutic treatment of rheumatic diseases.
5) In those cases of rheumatoid arthritis where hot-spring bathing was used in combination with prednisolone, excreted urinary 17KS decreased temporarily, but it tended to increase again a few days after the discontinuance of administration of prednisolone. There are many cases where a reduction in the amount of prednisolone to be administered will cause the appearance of reactivation symptom of rheumatoid arthritis, but it is anticipated that hot-spring bathing can prevent it to some extent, judging from the changes in excreted urinary 17KS caused by balneotherapy. Further examination is needed for any definite conclusion.

The author studied the effect of hot-spring bathing on the pituitary adrenocortical function of rheumatoid arthritis patients by means of Thorn's test, employing Nomin-no-yu and thermal water of the University Hospital (sulfated spring), and obtained the following conclusion.
1) Unusual count of eosinocytes can be found among persons who are apparently healthy, and who cannot be the good subject of Thorn's test. Such unusual cases were excluded. Thorn's test revealed that the reduction of eosinocytes were more than -50.0% (an average of -55.2%) in most of healthy persons (11 out of 15 cases).
2) The number of eosinocytes in blood of almost all healthy persons decreased through hot-spring bathing, showing a decrease of 15.8% on an average, which shows that hot-spring bathing has an action similar to that of ACTH. Such a phenomenon will be more pronounced with the initiation of balneotherapy, showing the rate of decrease as high as 35.3% on an average. Namely, the action of hot-spring bathing similar to that of ACTH tends to be intensified with the initiation of balneotherapy.
3) Thorn's test in rheumatoid arthritis patients revealed that the number of those in whom the rate of decrease was more than 50.0% was about equal to that of those in whom the rate of decrease was less than 50.0%, showing an average rate of 50.8%, which is almost similar to that of the healthy subject. In the case of those who had been suffering from rheumatoid arthritis more than a year, both the stage of progress of symptoms (stage) and the class of dysfunction (class) were high, and in such cases, the rate of eosinocyte decrease in Thorn's test was clearly less and the lowering of pituitary adrenocortical function was also observable.
4) It was clearly observed that Thorn's test on rheumatoid patients, when performed together with balneotherapy, showed an increase of the rate of eosinocyte decrease in the most of cases. At the same time it also increased generally the excreted urinary 17KS in those patients. Balneotherapy performed in combination with medicamenttherapy increased the rate of eosinocyte decrease in most cases. The increase was especially clear in those in whom the rate of decrease had been less than 50.0% before the initiation of balneotherapy. However, improvements of Thorn's test did not necessarily go in parallel with therapeutic effect and improvement in symptoms.
5) These findings show that the changes observable in Thorn's test provide us with a guide in balneotherapy of rheumatoid arthritis and are useful in explaining the balnectherapeutic effect in rheumatic disease.

The result of our general and medical survey are summarized as follows:
1) More than half the number of visitors to this spa are those who live in Yamagata Prefecture, and the rest are inhabitants of neighboring prefectures. Compared with the reports of our previous surveys, the number of inhabitants of neighboring prefectures is larger. Farmers are the largest in number, and their ages range from 20 to 60.
2) Considering the fact that more than half the number of spa visitors come here with therapeutic or convalescing purpose, this spa be said to be a spa where the sole object of visitors should be the hot spring curative treatment.
3) Form the viewpoint of classification of diseases, it may be said that patients with rheumatism and neuralgia are more than half the number, followed by those with the diseases of the stomach. This clearly shows the reason why this spa is called “the hot spring for rheumatism and neuralgia”.
4) More than half the number of spa visitors stay for about 2 weeks, and most of them take bath 5 to 6 times a day for curative treatment. Generally the spa visitors tend to bathe in excess.
5) 37.4% of the spa visitors drink hot spring water for curative treatment and this rate of drinking cure is higher than that in our previous reports.
6) Only 5.7% come to the spa under doctors' direction. 31.6% of them are conscious of the bathing reaction, symptoms of which are headache, feeling of weakness, anorexia, constipation, etc.
7) Observation of the effect of hot spring bathing on Thorn's test shows that in the larger number of patients with rheumatism and neuralgia the rate of decrease in eosinophilic leucocyte count becomes clearly higher after single bathing, but in healthy persons it is not uniform. And observation of changes in these rate of variations after succesive curative bathing reveals that in the group in whom the rate of decrease is low at the beginning of the bathing the vate tends to become higher after a week of curative bathing, while in the group in whom the rate of decrease is high it tends to become lower.

This paper deals with general survey of hot spring curative treatment at Higashine Spa, Yamagata Prefecture, and the effect of this spa on the capillary resistance and the variation in the blood pressure.
The results of the medical survey are summarized as follows:
1) Higashine Spa consists of saline weak common salt springs, in each of which chemical components are almost the same.
2) The visitors to this spa are inhabitants of Yamagata Prefecture, farmers being the largest in number.
3) Most of them (78%) are old men and women over 50 years of age, and the number of women are four times that of men.
4) The examination of spa visitors reveals that hypertension stands at the top. However, in most cases, the chief complaints are of rheumatic disorders in natwe and hypertension is found at the time of medical examination.
5) Those who bathe five times a day are the largest in number.
6) Those who noticed the bathing reaction are small in number. The bathing reaction may be considered as a serious thermal crisis.
7) Bathing in this hot spring increases the capillary resistance and lowers the blood pressure.
Accordingly, bathing in this spa, under a proper guidance, may be effective in the treatment of hypertension. For balneal treatment of hypertension, physicians' directions as to how and when patiens should bathe, will be necessary: unplanned bathing cannot be approved of.

Experimental and clinical studies upon the drinking effect of several hot springs were carried out.
Hot spring waters used were as follows;
(1) “MANJUYU”, being of H₂S-containing acid with pH 3.2, in Noboribetu.
(2) Simple thermal spring of KARURUS Spa, in Noboribetu.
(3) Muriated water of YUNOKAWA Spa, in Hakodate.
(4) Simple thermal spring of TOKATIGAWA Spa, in Obihiro.
Infant mice to which “MANJUYU” or simple thermal water of KARURUS were given, could not grow as well as control mice. Especially “MANJUYU” gave worse infulence than KARURUS to their growth.
The weight of the adrenals of adult mice increased evidently after drinking of “MANJUYU” and KARURUS spring for few weeks. However, as the time of death resulting from drowning was not prolonged, the resistance against stress did not seem to be significantly raised. Therefore it was thought that the hot spring drinking is not always profitable to animals.
Being given to the patients with gastric disorders, muriated water of YUNOKAWA improved the changes of HCl-secretion, juice-secretion and corrected disturbance of stomach movement.
To some patients it had poor infulence however. Beside, one must be careful not to forget that muriated water has a tendency to inhibit diuresis.
Drinking of simple thermal water of TOKATIGAWA has also similar effect to muriated water on gastric disorders. 1/4 of the patients with gastric disorders complained of nausea and vomitting.

The effects of thermal bath upon the sexual functions such as sexual circle and upon sexual centers such as diencephalon, hypophysis, etc. which control sexual circle were investigated.
1. It was observed by vaginal smear that the elongation of sexual circle by daily thermal bathing was chiefly due to the duplication of diestrus phase, and that the continuous estrus phase was temporarily interrupted by daily bathing on the 1st and 2nd days of bathing and between 10th-13th days. Further it was also ascertained by Hooker-Forbes test that the sensitivity of the uterus to progesterone was enhanced by daily bathing.
2. Castrated rats, which vaginal introitus were closed with calodium, were bathed in 38°C simple water bath containing estrogen for 10 minutes and praestrus or estrus phase were observed in the vaginal smears of these rats after bathing. This effect was more remarkable in accordance with higher estrogen content and in rats which back hair was cut off. The action of estrogen in the solution became almost unrecognizable when mixed in mineral mud. We consider it has a significant value as a new peculiar way of estrogen application to mix estrogen in simple water and let it absorb through total skin area which was induced to state easy to resorb by bath.
3. Simple water bath containing progesterone and androgen induced no definite changes in sexual circle. Continuous estrus phase showed on the 3rd-14th days of androgen containing and on the 3rd-12th days of progesterone containing baths in case of 1st phase deviation.
4. By simple water bath containing copper salt of minor quantity, vaginal smear circle showed marked decrease of diestrus phase and the increase of estrus or metestrus phase.
5. The urinary gonadotropin was decreased on the 3rd day of daily bathings in the most of cases and thereafter came back to the level before bathing or temporarily increased to higher level and then got back to the previons level.
6. Oxygen consumption of diencephalon became rather lower by simple water bath but almost unchanged by copper-sulfate-bath, and was lower after estrogen-bath.
0₂-consumption of the brain cortex showed slightly lower value after bathings of any kinds.
7. The changes of E. E. G. (electro-encephalogram) by daily bathings in several kind of thermal waters, in sexnal hormone containing water, and in copper sulfate containing water were investigated. But no physiological meaning was obtained from E. E. G.
8. The changes of urinary 17-KS and its fraction by bathing were investigated taking the value before bathing as standard, By daily bathing in simple water, 17-KS values fluctuated-significant rise during ca. 10 days and slight fall before and after the rise.
The fraction ratio of adrenocortical substances to substances of sexual glands slightly increased by bath. By daily bathing of simple thermal water, 17-KS showed marked increase on the 3rd and on the 10th day of bathing, and rapidly decreased to the level lower than the level before bathing after cessation of bathing. The fraction ratio described increased remarkably this time. By daily bathing in simple water containing estrogen, the increase of 17-KS following the initial decrease was obscured. The fraction ratio by this time showed no marked changes. By daily bathing in simple water containing copper sulfate, 17-KS increased lately. The fraction ratio increased remarkably.
9. We took oligomenorrhoe cases as a sample in gynecological application of thermal bath, and applied simple water bath containing estrogen, so-called estrogen bath, together with hormone therapy and found better improvement of smear index, fern test and cervical mucus, and the induced genital bleeding. Although estrogen bath itself had only slight effects, but it promoted the therapeutic effects of sexual hormone applied.
Next, as an application of thermal bath in tocology we chose partus inducement in terminal stage of pregnancy. We appli

Since 1933, effects of the internal use of mineral waters in this district were studied by Prof. Dr. T. Misawa and his coworkers concerning gastric and bile secretion, gastrointestinal motility, diuresis, glucose metabolism, anemia and others.
In this report, present status of the internal use of mineral waters in this district, the results of studies on the therapeutic and toxic effects, maximum permissible concentrations of trace elements in the spring waters, several experimental and clinical studies on atherosclerosis etc. will be reported.
I. The present status of internal use of hot spring waters in Kanto district and its environs.
The research was carried out by enquête method. Answers were obtained from 40spa admimistrations. Following results were concluded:
1) In the majority of the spas the internal use of spring waters are uncommon, only at 3 spas (Spas Shima, Kuronagi, Shibu) the internal use is more frequently carried on than the bathing cure.
2) The majority of the visitors take hot spring waters 2 times a day (morning and evening), ca. 100ml at a time, in total usually 5 go (900ml-1000ml), maximum 1.5 sho (2700ml), a day.
3) The majority of patients utilizing the drinking cure are suffering from gastro-intestinal diseases, and rarely from other diseases.
4) For the gastrointestinal disease, sodium chloride waters, sulfur waters and alkaline waters are used mostly and for the constipation hydrogen sulfide waters, sodium bicarbonated waters and sulfated waters were used.
II. Statistical observation on the spa treatment in Shiohara Hot Springs, Tochigi Pref. 1) The patients had predominantly gastrointestinal disorders in Motoyu (pH 6.3, hydrogen sulfide containing sodium bicarbonate and chloride spring).
In Motoyu, the patients with drinking cure suffered from stomach ulcer (15/41), chronic gastritis, duodenal ulcer and gastroptosis with atonia. It seems that they select a spring according to their empirical evaluations.
2) About 87% of the visitors put in practice of drinking of hot spring water in Motoyu. Most visitors take thermal bath 5 to 6 times a day on an average, and drink spring water at the time of bathing, in total 4-6 go (720-1080ml) a day in 18/41 cases, 9-10 go (1620-1800ml) in 12/41 cases.
3) Liver function of patients (B. S. P. test and Gros test) was investigated before and after the cure, but no significant change was observed.
III. Effects of trace elements.
1) Effects of the addition of Cu and/or vitamine C on the absorption of iron in mineral waters from the digestive tract were studied using the radioactive iron experimentally.
Iron absorption from the digestive tract was promoted by the addition of copper, and vitamin C in the test solution. No promoting effect was proved by the addition of copper in the ratio of Fe:Cu=1:1, but it proved most effective with the ratio of
Fe:Cu=2.6:1
2) Maximum permissible amount of Pb, Zn and Cu contained in mineral waters were studied in rabbits experimentally. It was concluded that the maximum permissible dose of orally given Pb, Zn and Cu were between 7.5mg, 20mg or 1.9mg per kg body weight daily by administration for a month, respectively.
This concentration of lead is 100 times higher than that of mineral waters with highest lead content in Japan. And as mineral waters with high zinc or copper content in Japan used to be strongly acid and mostly belong to acid vitriol or acid alum vitriol springs, only daily 10-50ml can be taken after 5-10 times diluted with common water for drinking cure.
The highest record of zinc content in Japan is 965mg/kg ZnSO₄ (Sasakura Hot Spring), therefore there would be no probability of Zn poisoning from drinking cure of mineral waters in Japan, and when taken 25ml of copper containing mineral water daily the maximuan permissible concentration of copper containing waters will be 75mg/kg. Only few of the Japanese mineral waters contain more than