To protect renal parenchyma from ischemic damage during pedicle clamping for renal surgery, renal cooling by using in situ renal perfusion was performed. Fifty-six mongrel dogs which were unilaterally nephrectomised three or four weeks before were anesthesised. A KIFA red catheter was placed in the left renal artery and a KIFA black catheter was placed in the left renal vein, via the femoral vessles by Seldinger's method. The animals were divided into the following six groups and the kidneys were perfused.
Group I, control. Group II, normothermic ischemia. Group III, intermittent perfusion with 4°C cooled hyperosmoral lactated-Ringer's solution. Group IV, intermittent perfusion with 4°C cooled Sack's solution. Group V, continuous perfusion with Fluosol-43 at room temperature. Group VI, continuous perfusion with cooled Fluosol-43.
Two series of experiments were performed. In the first experiment renal function immediately after 90 or 180 min, of ischemic period with or without perfusion was determined. In the second experiment renal function was followed during recovery period from ischemia. RBF, Ccr, PSP excretion test, U/P osmolar ratio and GOT, GPT levels were evaluated in each dog. The left kidney was removed for histological studies.
The following results were obtained.
1) No urine production was found in Group II, but in Groups from III to VI, urine was obtained. However, renal function immediately after perfusion was reduced extremely.
2) The elevation of serum creatinine in the perfusion group was minimum. Serum creatinine level of one animal which belongs to Group III was elevated to 5.4 mg/dl on the third day after perfusion.
3) Renal function recovered to the normal range on the seventh day or the eighth day after perfusion except in group III.
4) Intracellluar hyperosmoral “Sack's” solution was the most suitable perfusate for in situ renal perfusion.
5) Renal function on the seventh or eighth day after perfusion was good in animals perfused with Fluosol-43 at room temperature.
6) The in situ renal perfusion would have definite advantages in certain renal procedures such as partial nephrectomy, renal artery replacement and certain cases of nephrolithotomy.

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A quantitative study was carried out on the lactate dehydrogenase (LDH) and its isozymes in ejaculates of 170 men, testicular tissues of 12 men and accessory sexual organs of 10 men. LDH activities were assayed on the basis of the rate change of NADH concentration during the convertion of pyruvate to lactate at 37°C by measuring light absorbance at 340nm spectrophotometrically. LDH isozymes were separated by agar-gel electrophoresis. The isozymes were then visualized by incubating the gel in a buffer solution containing blue tetrazolium salts, phenazine methosulfate, NAD and lactate as substrate. The relative proportions of the isozymes were estimated by scanning the stained gel at 570nm in Densitron 1M (Joko & Co.).
The LDH activity of the normospermia (n=16) was 5280±3217mU/ml (mean±S. D.) and the mean % distributions of the enzyme activity among isozymes were 3.9±1.6% for LDH₁, 14.7±3.3% for LDH₂, 24.0±4.3% for LDH₃, 21.6±11.1% for LDHx, 20.9±5.6% for LDH₄ and 14.6±7.3% for LDH₅.
The LDH activity and sperm count revealed significant correlations (n=1; 0, r=0.50, p<0.001). But there was no significant correlation between the enzyme activity and sperm motility. The LDHx was recognized in 96.6% of ejaculates with sperm counts above 50 million/ml, in 100% of those with sperm counts from 20 to 50 million/ml, in 93.5% of those with sperm counts between 20 and 10 million/ml, in 52% of those with sperm counts under 10 million/ml and in 0% of azoospermia.
There appeared to be a tendency that the higher the sperm motility in the ejaculates, the higher the incidence of the presence of LDHx in ejaculate.
Both LDH₁ and LDH₂ showed no relations to sperm counts. While activities of LDH₃ (r=0.38, p<0.01), LDH₄ (r=0.42, p<0.01) and LDH₅ (r=0.27, p<0.01) respectively revealed significant correlations to sperm counts. Especially LDHx activity revealed a high correlation to sperm counts (r=0.60, p<0.01).
The activity of each isozyme including LDHx showed no significant correlation to sperm motility.
After vasectomy, LDHx fraction disappeared from seminal plasma and total LDH activity was significantly reduced to 2901mU/ml from 3838mU/ml (p<0.05).
Six patients with varicocele who had demonstrated oligospermia with lacked LDHx in the ejaculates underwent high ligation of spermatic vein. LDHx was recovered in seminal plasmas of 5 of them with improved sperm counts.
Fresh spermatozoa were washed with physiologic saline 4 times until almost no LDH activity was detected in the solution used. And then the sperm suspension was kept standing at room temperature for 144 hours. The LDH activity was again recognized in the fluid of the suspension and consisted of only LDHx.
LDHx was detected in the cytoplasma of testicular tissues (9 of 10 testes) and epididymal head (1 of 10 epididymides examined). No LDHx was found in any other tissues of accessory sexual organs examined.
In conclusion, LDH activity and LDHx fraction in human ejaculate would be a good index for clinical evalution of the ejaculate.

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Using the experimental apparatus for microexplosion, the peak pressure of blast wave which indicated dynamic effect of lead azide explosives was measured in the four directions (0°, 45°, 90°, 135°) to the central axis of charging chamber and at various distances from its front.
The results were as follows:
1) The scaling law already established in general explosion was confirmed also in microexplosion.
2) The isobarlic profile of the peak pressure showed that the blast wave propagated to the anterior direction of explosive charging chamber, mostly to the central axis of it.
3) The following experimental formulas relating to peak pressure and scaled distance in the four directions (0°, 45°, 90°, 135°) were obtained:
(i) P_0°=2.5×10⁴(³√W/D)^2.8
(ii) P_45°=4.2×10(³√W/D)^1.2
(iii) P_90°=1.2×10(³√W/D)^1.1
(iv) P_135°=2.0(³√W/D)^0.6
P: Pressure (kg/cm²)
W: Weight (g)
D: Distance (cm)

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For the purpose of seeking an optimal administration schedule of antimicrobial agents, therapeutic effects on experimental infection in mice were studied. Groups of ten mice were infected intraperitoneally with E. coli and treated with intramuscular injections of ampicillin or kanamycin.
In order to compare the therapeutic effects with two different dosage schedules, one group (Group A) was injected a definite dose of those antibiotics only once 1 hour after infection and another group (Group B) was injected twice every one half of the definite dose 1 and 5 hours after infection. The therapeutic effects on the group B was superior than that on group A with ampicillin, but kanamycin showed the almost same effects on groups A and B. Thus, concerning the patterns of blood level of drug obtained by two different schedules of injection, with ampicillin, it seemed to be more important to keep the effective blood level of drug long than to increase the drug concentration transiently. On the other hand, as for kanamycin, high concentration of the drug was also important in order to maintain the effective blood concentration of drug.
The therapeutic efficacy when a definite dose of ampicillin was divided equally into five and each of them was injected to mice 5 times hourly was less superior than that when the dose of the initial injection of the 5 injections was increased, decreasing the doses of the subsequent 4 injections. The results suggested that in order to get the therapeutic effect effectively, the initial drug concentration was especially important.
The results obtained by these animal experiments were consistent with those of the already reported in-vitro experiments.

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The effect of chemotherapy on experimental pyelonephritis in rats was investigated by initiating treatment at various intervals after induction of the disease. An experimental model of pyelonephritis was produced by transurethral inoculation of Proteus mirabilis 24 hours after producing chemical cystitis by instilling 0.5ml of 0.75% acetic acid into bladder transurethrally. The bacterial inoculum was 0.5ml of heart infusion broth containing 10⁶ cells/ml of P, mirabilis isolated from a 33-year-old woman with bilateral chronic pyelonephritis and renal stones. The animals were administered 100mg of ampicillin per kilogram of body weight intramuscularly twice daily for 7 days from 1, 3, 5, 7 and 10 days after inoculation. At 4 weeks after inoculation, urinary concentrating ability, renal histology, serum antibody titer and urinary tract stone formation were examined.
The results obtained were as follows.
1) The incidence of pyelonephritis with this experimental model was 94.0%, including rats that died in the observation period. In controls not pretreated with acetic acid prior to P, mirabilis inoculation, the incidence of pyelonephritis was 71.7%. This difference was significant (p<0.01). Serum antibody titer measured by bacterial agglutination displayed a dramatic rise from 1 to 2 weeks after inoculation and increased gradually thereafter.
2) Disturbances of urinary concentrating ability were mildest in rats in which treatment was initiated 1 day after inoculation. The longer the interval between inoculation and the commencement of treatment, the more severe were disturbances of urinary concentrating ability.
3) Histological changes of the kidney were mildest in rats in which treatment was initiated 1 day after inoculation. The longer the interval between inoculation and the commencement of treatment, the more pronounced were histological changes.
4) Serum antibody titer was low in each group in which treatment was initiated within 7 days of inoculation compared to that of nontreated group.
5) A significant correlation between impaired urinary concentrating ability (urinary osmolarity after withdrawing all water for 24 hours) and serum antibody titer was seen in each of group with the exception of that in which treatment was initiated 1 day after inoculation.
6) There was no stone formation of urinary tract in rats in which treatment was initiated 1 and 3 days after inoculation.
The results of this experiment indicate that the prompt treatment of pyelonephritis by suitable chemotherapeutic means is of vital importance as suggested by renal function, renal histology, and serum antibody titer. Furthermore, urinary concentrating ability and serum antibody titer were considered to be important parameter in the diagnosis and treatment of pyelonephritis.

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The present assay for oxalate in urine is a modification of the radioenzymatic isotope dilution technique developed by D. J. Bennett. This assay has become sensitive and reproducible by the use of a new CO₂ trapping apparatus. The oxalate in 2ml of urine is precipitated with Ca⁺⁺, dissolved in a citrate buffer. The oxalate is then decarboxylated with oxalate decarboxylase in the presence of approximately 405ng of [¹⁴C] oxalic acid (18.0nCi) to evolve labeled and unlabeled CO₂. An equation derived by Newsholme and Taylor describes quantitatively the simultaneous effects of the variation in the non-radioactive substrate on the isotope dilution and the enzyme velocity, resulting in a linear standard curve. With the extraction procedure 70% to 90% of the [¹⁴C] oxalic acid internal standard added to urine was recovered and the recoveries of added unlabeled oxalate averaged 103.5±2.1% (SE). The Km for this enzyme was found to be 35.6μM. The KI for phosphate and sulfate, which are competitive inhibitors of the enzyme, was found to be 0.9mM and 1.3mM respectively. The sensitivity of this method was 0.5μg, which is close to the theoretical sensitivity of 0.1Km, The coefficient of variation (CV) for triplicate determinations of triplicate precipitations of urine was 9.3%. The 24-hr urine excretion of oxalate from 20 patients who had normal kidney function without urinary stones or metabolic deseases, was 24.4±6.9mg (Mean±SD).

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There is ample evidence that the compensatory secretion of adrenal steroids occurs after estrogen administration and castration as the treatment for prostatic carcinoma. Thus, the adrenal androgens dehydroepiandrosterone (RHEA), dehydroepiandrosterone sulfate (DHEAS) and 4-androstenedione have been postulated to facilitate the growth of prostatic carcinoma, despite the loss of testicular function by estrogen therapy.
Based on this, pituitary ablation, corticosteroid therapy and adrenalectomy have been established as therapeutic programs for relapsing prostatic carcinoma. However, it is still controversial whether the synthesis of adrenal androgens is stimulated concomitantly with hypersecretion of corticosteroid resulted from hyperplasia of the adrenal cortex following estrogen therapy and castration. Furthermore, it is also unknown whether the adrenal androgens trigger the reactivation of prostatic carcinoma.
For the purpose to elucidate the problems mentioned above the evaluation of the basic androgen milieu in patients with prostatic carcinoma treated with estrogen and gestagen, and the determination of 4-androstenedione concentrations in peripheral plasma by a specific radioimmunoassay in several clinical conditions of prostatic carcinoma were employed as the experimental design for the studies.
The results in the present paper were summarized as follows:
1) Plasma 4-androstenedione levels in normal men, normal women and normal aged men were 81±38 (M1±SD, n=10), 105±22 (M±SD, n=10) and 73±28 (MSD, n=9), respectively. Similarly, the androgen levels in benign prostatic hypertrophy and prostatic carcinoma were 74±22 (M±SD, n=21) and 52±25 (M±SD, n=9), respectively. In the plasma 4-androstenedione level, there is no statistically significant discrepancy between benign prostatic hypertrophy and prostatic carcinoma in the comparison with normal aged men. Furthermore the concentration of 4-androstenedione in the morning was 20 per cent higher than that in the afternoon.
2) The testosterone levels in 15 patients treated with 30mg daily of hexestrol for 1 to 8 years were 34.5±19.7 (M±SD). The low level of plasma testosterone was maintained with estrogen treatment at least up to 8 years.
3) The formation of testosterone from progesterone³H- in the testicular tissue in vitro was definitely inhibited by oral administration with more than 30mg daily of hexestrol, 3mg daily of diethylstilbestrol or more than 200mg daily of diethylstilbestrol diphosphate.
4) The plasma 4-androstenedione in the patients treated with 30mg daily of hexestrol for 1 to 9 years was 52.9±20.0 (M±SD, n=29). No patients who have the plasma 4-androstenedione over the normal level were recognized during the estrogen therapy, while plasma cortisol level following estrogen therapy was three times higher than the normal level. It is also observed that the plasma androgen in the patients treated with 100 mg daily of chlormadinone acetate for 6 years remained within normal level.
5) The plasma 4-androstenedione concentrations in 8 relapsing prostatic carcinoma were within the range of normal aged men.
It is reasonably concluded that there is no tendency to stimulate the synthesis of adrenal androgen in the long term treatment with estrogen or chlormadione acetate, and that the adrenal androgens particularly 4-androstenedione can not become a trigger for reactivation of prostatic carcinoma.

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Human prostate has been known to contain a high concentration of zinc by histochemical and biochemical studies. Though zinc seems to play an important role in the prostate, little was known on the site and mode of action of the prostatic zinc. The object of the present study is to demonstrate the ultrastructural localization of zinc in the hyperplastic, as well as neoplastic prostate using sulfide silver method, and to determine the content of the reaction products in the ultrathin section by X-ray microanalyzer.
Specimens were fixed in 2% glutaraldehyde saturated with hydrogen sulfide at 4°C for 2 hours. After ultra-thin sections were mounted on an Au grids, they were developed in the Timm's developing solution for 5 to 30min. Control studies were carried out with non-developed and ungassed sections. Developed and undeveloped ultra-thin sections were observed with JEM 100CX with energy dispersive type X-ray microanalyzer.
Abundant reaction products were observed in the secretory vacuoles of the hyperplastic prostate. Aggregations of the silver precipitation were clearly seen on the vesicles of the secretory vacuoles. None of the deposits was detected on the nucleus, lysosomes, Golgi apparatus and other cytoplasmic organelles. These findings were also observed in the neoplastic cells. Undeveloped but fixed with sulfide sections showed moderate changes ultrastructurally compared with conventional section. Small amounts of electron dense materials were noted in the secretory vacuoles, while the nucleus and other cytoplasmic organelles remained unchanged. By an X-ray microanalyzer, reaction products of the developed section were detected to contain a high amount of silver and sulfate. X-ray microanalyzer could reveal K_α and L_α of zinc at the electron dense material in the secretory vacuoles of the undeveloped section.
From the present investigation, prostatic zinc was clarified to localize in the secretory vacuoles of the prostate.

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In order to detect the effect of hyperthermia on urinary bladder cancer, in vitro studies were done using two human bladder cancer cell lines (T-24, MGH-Ul: Kindly supplied by Prof. G. R. Jr. Prout, Harvard Medical School, Boston) and HeLa cells. Hyperthermic treatment at 43°C for one hour resulted in the same amount of cytotoxicity and inhibition of cell growth in all three cell lines. A combined treatment of hyperthermia and bleomycin caused more synergistic cell killing than that seen with hyperthermia alone.
Those results indicate that hyperthermic treatment with or without bleomycin is effective in killing cultured bladder cancer cells and suggest that hyperthermia may be a useful approach in clinical management of cancer of the bladder.

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The kinetics of cell killing by hyperthermia alone and the mechanism of the synergistic cytotoxicity of hyperthermia plus bleomycin were studied using HeLa cells.
Investigation of the kinetics of cell division after hyperthermic treatment (43°C, 45min) indicates that there exist two cell subpopulations, one which is no longer capable of further growth and another which regains growth capacity after a lag period.
Combined use of hyperthermia and bleomycin inhibited the recovery of the growth capacity in this second subpopulation of cells. Hyperthermia alone did not cause the breakage of DNA-single strands as determined by the alkaline sucrose gradient technique. Combined use of hyperthermia and bleomycin did, however, produce more DNA single strand breaks than bleomycin alone. This combined treatment was also found to inhibit the repair of single strand DNA breaks by bleomycin.
The combined use of bleomycin and hyperthermia did not produce a synergisticinhibition of the HeLa cells' DNA, RNA, or protein synthesis.

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In order to develop a new therapy for the management of bladder cancer, hyperthermic therapy of bladder cancer using bladder irrigation with warmed saline, was investigated. Thirty-four patients with multiple transitional cell carcinoma of the bladder were treated with 4 kinds of hyperthermic therapy; hyperthermia alone, combined therapy of hyperthermia and bleomycin, combined therapy of hyperthermia and radiation, and combined therapy of hyperthermia, bleomycin and radiation. Hyperthermic treatment was accomplished as described below: warmed saline was irrigated into the bladder cavity through a 3-way catherter with the temperature in the bladder being maintained at 42°-43°C. A course of treatment consisted of a series of 10 to 12 applications of hyperthermia for 1 hour each. For those patients receiving hyperthermia and bleomycin, bleomycin was added to saline solution to a final concentration of 30mg/l. In those treatments involving radiation, 150 to 200rad/day were administrated to the bladder externally using Linac of ⁶⁰Co. Immediately after irradiation, patients were irrigated with warmed saline (with or without bleomycin) for 1 hour. The total dose of the radiation was limited to 3, 500 to 4, 000rad. Obvious tumor regression was observed in two out of six patients treated with hyperthermic therapy alone. Similar regressions were observed in one out of four, five out of nine and twelve out of fifteen patients who were treated with combined therapy of hyperthermia and bleomycin, combined therapy of hyperthermia and radiation, and combined therapy of hyperthermia, bleomycin, and radiation, respectively.
Among the twenty-four patients who received the combined therapy of hyperthermia and radiation (with or without bleomycin), low staged tumors (T1) responded most effectively to the therapy. Significant results were, however, also obtained with T2-T3 staged tumors. High grade tumor (G2-G3) showed an increased response over low grade tumor (G1).
The most frequent side effects of those therapies were bladder irritation and urethral pain. Bladder irritation was induced most intensively by the combined therapies of hyperthermia, radiation and bleomycin. This side effect was reversed except in one patient. There were no serious complications.
Immunological competency of the patients treated with combined therapy of hyperthermia and radiation (with or without bleomycin) was examined using parameters such as lymphocyte counts and PHA, ConA blastogenesis of peripheral lymphocytes. This therapy induced suppression of blastogenesis of lymphocyte. But this effect was quantitatively the same as that of radiation therapy alone on the bladder cancer.
Those results suggest that hyperthermic therapy is clinically applicable for the treatment of bladder cancer. Those treatments may be particularly useful in the case of multiple superficial tumors, as an adjuvant treatment of preoperative radiation with cystectomy, and in those patients with severe complications of the lung, heart and others.

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We performed preoperative transcatheter parasitic arterial embolization using Gelfoam in one patient with left renal adenocarcinoma of clinical stage T3. The parasitic arteries consisted of the left renal artery, left phrenic artery and left colon branch of the inferior mesenteric artery. Subsequently, tumor regression was noted on CT and abdominal palpation 7 days after the embolization. The side effects were gastrointestinal symptoms, weakness of intestinal peristalsis, left frank pain, pyrexia, leucocytosis and elevated serum GOT. GPT. LDH, but all these clinical findings were transient.

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Case 1: A 57-year-old female, complaining of fever and pain in the left flank, was admitted. A diagnosis of left pyelonephritis was made, but the patient gradually developed a bulging in the left flank. A KUB film revealed a soft-tissue mass inferior to the left kidney. An excretory pyelogram showed that the middle and lower cayces of the left kidney were not sharply outlined, and that the urinary bladder was displaced to the right. X-rays of the chest revealed a collection of fluid at the left base. A diagnosis of left retroperitoneal space abscess was made, and the abscess was drained. Cultures of the pus showed do presence of bacteria.
Case 2: A 48-year-old female, complaining of pain in the left flank and a high fever, was admitted. The patient had previously had an appendectomy fifteen years before. A diagnosis of left pyelonephritis and left ureteral stone was made, but she suddenly developed severe dyspnea. X-rays of the chest showed effusion. Later, physical examination revealed a tender mass in the right lower abdomen. An excretory pyelogram revealed a large homogeneous density, which corresponded with the palpable mass, and the upward displacement of the right kidney. First the right and then the left retroperitoneal abscesses were drained. Cultures of the pus revealed no presence of bacteria. In both cases 1 and 2, left pyelonephritis developed into left retroperitoneal abscess, and the left abscess in case 2 was thought to have spread further into the right retroperitoneal space through the pelvic space. The hydrothorax was thought to be secondary to retroperitoneal space abscess, because it disappeared simply by drainage of the abscess.
The records of 62 cases of retroperitoneal abscess during the last 20 years reported in Japan were reviewed, and the causes, spread of infection, and development of hydrothorax in retroperitoneal abscess were discussed.

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