In Vitro metallic mercury uptake by rat liver dissociated cells, homogenate, cell components (microbody-rich fraction, mitochondrial fraction and microsomal fraction), heme protein (catalase, cytochrome c, methemoglobin and hematine) and ferric ion were investigated. The effects of ethyl and methyl alcohol on the uptake were also investigated.
The following results were obtained.
1) Mercury uptake by rat liver dissociated celld was similar to the uptake by homogenate.
2) Mercury uptake by rat liver homogenate in the presence of hydrogen peroxide was 1.22 times the uptake in the absence of hydrogen peroxide. The uptake by rat liver dissociated cells in the presence of hydrogen peroxide was similar to the uptake in the absence of hydrogen peroxide.
3) Mercury uptake by crystalline catalase, rat liver dissociated cells and homogenate was inhibited by ethtl alcohol and methyl alcohol, though inhibition of catalase activities was not observed.
4) Mercury uptake by rat liver cell components was observed in the order of microbody-rich fraction>microsomal fraction>mitochondrial fraction, and catalase activities were also observed in the order of microbody-rich fraction>microsomal fraction>mitochondrial fraction. Mercury uptake by rat liver cell components was inhibited by potassium cyanide and azide.
5) Cytochrome c, methemoglobin, catalase and ferric ion oxidized metallic mercury in the presence of hydrogen peroxide. In the absence of hydrogen peroxide, methemoglobin and ferric ion oxidized, but catalase and cytochrome c did not oxidize metallic mercury.
6) Hematine did not oxidize metallic mercury with or without hydrogen peroxide.
7) Uptake by ferric ion was inhibited by equimolar addition of EDTA.
8) In the presence of hydrogen peroxide, mercury uptake by catalase was 27, 000 times, cytochrome c was 140 times and methemoglobin was 35 times the uptake by ferric ion. Therefore, of heme protein catalase showed the highest mercury uptake.

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The quantity of Phenylsulfate(PhS) and Phenylglucuronide(PhG) excreted by rats after an oral dose of 1m mol/Kg phenol was estimated and the following results were obtained.
1. Both PhS and PhG were almost completely excreted within 1 day.
2. The excretory quantity of PhS was greater than that of PhG.
3. High performance liquid chromatography, simplified the determination of PhS and PhG.

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B-cell and T-cell human leukemic cell lines, BALL-1 and TALL-1, were transplanted into newborn hamsters immunosuppressed with rabbit anti-hamster lymphocyte serum. The growth inhibitory effect of interferon (IF) was tested by intraperitoneal administration in these tumor-bearing animals. The following results were obtained: (1) TALL-1 transplanted intraperitoneally was inhibited much more than BALL-1 transplanted subcutaneously. The life span of hamsters transplanted with TALL-1 was also prolonged. These findings coincided with the results of in vitro sensitivity of these two cell lines to IF. (2) The antiproliferative effect of a combination of cyclophosphamide and IF to BALL-1 was greatly enhanced compared with when either agent was used alone. (3) The combined use of L-asparaginase and IF was similarly more inhibitory to TALL-1 than either agent alone. It seems that the growth of residual tumor cells after chemotherapy can be effectively inhibited by IF. In using IF for cancer chemotherapy, therefore, it would be advisable to combine chemotherapeutic agents with IF.

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Physical loading can be monitored by measurement of the amounts of urinary protein. The Bio-Rad method in combination with gelfiltration gave the following results.
1. The presence of albumin and γ-globulin in proteinuria was recognized by immunodiffusion in antigen-containing gel. The presence of α₁-Acid-Glycoprotein, Transferrin and Zn-α₂-Glyco-protein was also demonstrated. However, other proteins need to be confirmed.
2. With the direct Bio-Rad method, some low molecular substances other than protein showed coloration. Urinary protein over 4000 molecular weight could be measured by removing lower molecular weight substances by gelfiltration. (Fil-Col method).
3. Small amounts of urinary protein could be determined by the modified turbidimetric method of Meuleman. Good correlation was obtained between the Fil-Col method and the turbidimetric method, and between the Fil-Col method and the TONEIN-TP method.
4. From the above results, it was concluded that the Fil-Col method is useful for the exact quantitative determination of minute amounts of urinary protein, and that the TONEIN-TP method is useful for screening of minute amounts of urinary protein.

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Patients with acute leukemia are prone to get infections. Pathological conditions such as leukocytopenia and disturbances of humoral and cellular immunity are thought to contribute to this high susceptibility to infection.
Superoxide, one of the active oxygens, is thought to be related to the intracellular bactericidal activities of neutrophils, particularly in oxygen dependent systems. These findings led the author to speculate that primary and secondary disturbances in the superoxide production of neutrophils are of importance in the high susceptibility of patients with acute leukemia. In this experiment, the superoxide production of normal human neutrophils stimulated by Concanavalin A and Cytocharacin D was assayed by spectrophotometer in order to evaluate its usefulness in clinical applications. The effect of age and sex on supreoxide production was also examined. The following results were obtained:
1. The activity of superoxide production was affected by both time and temperature during cell preservation. It was also dependent on the cell concentration, temperature and pH during stimulation of the neutrophils. The optimal system for assay of the superoxide production of human neutrophils was as follow: Cells should be preserved at 0°C within 60 minutes. During stimulation, cell concentration, temperature and pH should be ajusted to 0.2-0.5×10⁶/ml, 37°C and 7.4, respectively.
2. There were no significant changes in superoxide production between males and females. Superoxide production in older subjects(70-89 y/o) was significantly lower than that in younger subjects(20-69 y/o)(p<0.001). The values were 4.72±1.17 nmol/min/10⁶-PMNs in the former and 6.45±1.33 nmol/min/10⁶PMNs in the latter.
Assays of superoxide production of human neutrophils can be clinically used because of its simplicity and reproducibility. It was apparent that the assessment of superoxide production of neutrophils by age was necessary in clinical applications.

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It is well known that patients with acute leukemia have a predisposition for life threatning infections. Among several factors which disturb host defense mechanisms, quantitative and qualitative disorders of neutrophils are thought to be important. Up to date, there have been some papers concerning the bactericidal activities of neutrophils obtained from patients with acute leukemia; however, little information about the production of active oxygens, especially the superoxide anion, is available. Superoxide of neutrophils is one of the active oxygen related to intracellular bactericidal activities; therefore, in this study, superoxide production of neutrophils in patients with acute leukemia was examined in order to clarify the cause of the high susceptivility to infection. The results were as follows:
1. Before antileukemic chemotherapy, superoxide production was 4.09±1.43 nmol/min/10⁶PMNs in patients with acute leukemia. It was 6.45±1.33 nmol/min/10⁶PMNs in normal subjects. During induction chemotherapy, superoxide production was suppressed to a level of 3.56±1.18 nmol/min/10⁶PMNs; however, it recovered to normal range and somewhat overshot in the complete remission stage.
2. An inverse correlation between superoxide production of neutrophils and the total number of leukemic clls in the bone marrow was recognized.
3. The changes in superoxide production throughout the entire course were not correlated with the type of leukemia.
These results clearly indicate disturbance of superoxide production by neutrophils in patients with acute leukemia. They also suggest that alterlations in the intracellular bactericidal activities of neutrophils may be one of the reasons for the high susceptivility to infections of patients with acute leukemia.

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Previous work, described in part 2 of this study, showed that the superoxide production of neutrophils was suppressed by induction chemotherapy. These findings suggest direct effects of antileukemic agents on intracellular bactericidal activity; however, there is little information available concerning this aspect. The superoxide of neutrophils is one of the active oxygens and it is thought to be one of the major substances related to the intracellular bactericidal activities of neutrophils. This experiment was conducted in order to evaluate the direct effects of antileukemic agents on superoxide production of human neutrophils.
The following results were obtained;
1. The effects of antileukemic agents on superoxide production were examined before and after intensification chemotherapy in 14 patients with complete remission. Significant suppression of superoxide production was found 60 minutes after initiation chemotherapy; however, it recovered 24 hours after chemotherapy.
2. In the in vitro system, neutrophils were incubated with antileukemic agents for 3 minutes prior to stimulation by Concanavalin A and Cytochasin D. Among the antileukemic agents examined, Methotrexate, Vincristine and Prednisolone inhibited superoxide production with a concentration dependency. The 50% inhibition concentrations of these agents were 3.6×10^-4mol for Methotrexate, 3.2×10^-4mol for Vincristine and 4.2×10^-4mol for Prednisolone. Cytosine arabinoside, Neocartinostatine and Cyclophosphamide showed no inhibitory effects on superoxide production.
3. The inhibition of superoxide production by Methotrexate could not be reversed by Folinic acid in the medium.
4. In the cell free superoxide production system consisting of Xanthine plus Xanthine oxidase, Methotrexate and Prednisolone inhibited superoxide production with a concentration dependency. The 50% inhibition concentrations were 1.3×10^-3mol for Methotrexate and 2.4×10^-3mol for Prednisolone.
5. In the in vitro system, Methotrexate and Prednisolone were not only inhibitors of superoxide production but also scavengers of superoxide.
These results suggest that some antileukemic agents directly inhibit intracellular bactericidal activities, resulting in the susceptibility to infection of patients with acute leukemia.

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The antitumor effects and influence on cell-mediated immunity of Corynebacterium parvum (C. parvum) were studied using C3H/He mice and syngenic mouse ascites hepatoma MH 134 tumor.
With the administration of C. parvum alone, a significant antitumor effect was obtained in the group given 0.4mg per day intraperitoneally for 5 consecutive days from the fifth day after transplanting 5×10⁵cells/0.1ml of MH 134 tumor subcutaneously on the back. A significant antitumor effect was also obtained when C. parvum was combined with MF chemotherapy (Mitomycin C 1mg/kg, 5-Fluorouracil 25mg/kg), in the group given MF i.p. on the fifth day and 0.4mg/day of C. parvum i.p. for 5 consecutive days from the seventh day after tumor transplantation.
The cell-mediated immunity in these two groups was investigated by using the delayed type hypersensitivity (DTH) and macrophage migration inhibition factor (MIF activity). The results were: no decrease in DTH was shown compared with the tumor-bearing untreated group. MIF activity was marked compared with the tumor-bearing untreated group and tended to persist for a long period. From these findings, it was concluded that C. parvum was an effective immunopotentiator which yeilded an antitumor effect by restoring the cell-mediated immunity of the tumor bearing host. The effect seemed to be enhanced when used in combination with chemotherapy.

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A fecal examination for each of 471 residents of Manabe island was performed to clarify the status of parasitic infection by direct smear in August, 1977. The results were as follows:
1. Helminths found includedi Ascaris lumbricoides (2 cases, 0.4%), Trichuris trichiura (18 cases, 3.8%), Ancylostoma duodenale (5 cases, 1.1%) and Heterophyes heterophyes (4 cases, 0.9%).
2. The average infection rate was 6.2% for the whole area.

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Rabbit xenoantiserum was produced against a human null cell type leukemia cell line (NALL-1), and immune IgG was purified. NCS, a unique antitumor antibiotic that works on the tumor cell surface, was reacted with anti-NALL-1 rabbit IgG, in the presence of water soluble carbodiimide (WSCD). The resulting mixture was chromatographed on a Sephadex G-200 column. The first and second fractions were shown to contain NCS-immune IgG and no free NCS by the Ouchterlony double diffusion method and by immunoelectrophoresis. The conjugates inhibited the growth of Sarcina lutea and were more stable at conditions of 37°C or 56°C than free NCS. The conjugates did not loose inhibitory activities against the growth of S. lutea after frequent freezings and thawings. NCS-immune IgG might be useful in treating patients with cancer.

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The effect of microwave radiation (2450 MHz) on fungi, bacteria and viruses was examined to determine its applicability as a handy method of sterilization. Fungi (C. albicans) and bacteria (E. coli, P. aeruginosa, S. aureus and B. subtilis) stuck on the inside of a glass or plastic milk bottle were sterilized within 5 minutes of radiation regardless of whether the bacteria were dry or wet. Spores of B. subtilis and virus (ECHO virus and Herpes virus) were inactivated within 10 min without affecting the plastic or glass bottle. Wet condition rendered the micro-organisms more susceptible than dry conditions. The results indicate that the microwave oven may be suitable for sterilization of things such as feeding bottles or trays in infant rooms of hospitals.

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Blood vascular beds of the rhesus monkey kidney glomerulus were replicated with low viscosity methacrylate resin and observed in a scanning electron microscope.
The glomerular blood vascular beds consisted of freely and multiply anastomosing capillary networks, which showed lobulations. The lobules intercommunicated with each other by a few bridging capillaries. The lobules, however, could be recognized as fairly independent units in the glomerular microcirculation as the bridging vessels were small in size.
Very faint capillaries were sometimes observed in and near the vascular pole of the glomerulus. These faint vessels represented new formations of the glomerular capillaries.
Neither short-cut communication nor main pathways were noted between the glomerular afferent and efferent vessels nor within the glomerular capillary networks.

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Variations in the amount of Japanese Encephalitis (JE) virus in the field and outbreaks of JE were examined. The causes of changes in outbreaks were examined. The following results were obtained.
1. The level of JE virus estimated by positive rates of HI reaction and the 2-ME reaction of JE of swine tended to decrease from 1972 to 1977, and tended to increase from 1978 to 1980. On the other hand, positive rates of 2-ME reaction in 1978-1980 were higher than those in 1965-1971.
2. The number of JE-patients was more than 50 from 1965 to 1967, thereafter it decreased and no JE-patient was detected from 1971 to 1977. Thereafter an outbreak occurred again. Two definite cases and one suspected case were recognized in 1978, one suspeted case in 1979, and one definite case in 1980.
3. One cause of the decrease in JE-patients is that the size of a pigsty has been increased and sanitary conditions have been improved.
4. Discussion was made that the course of the outbreak of JE-patients from 1965 to 1980 was attributed to the amount of JE virus in the field estimated by antibody reaction in swine, the number of Ct, the arrangement of pigsties, the spread of window setting nets, improvement of nutrition, decrease of overwork and ruralization.

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