Search

Carbonic anhydrase activity in mouse kidney, liver and blood samples prepared from several methods were determined by both manometric (Altschule and Levis' method) and colorimetric (Maren's and Wilbur and Anderson's methods) methods with the following results.1. Between carbonic anhydrase activities of kidney, liver and blood and logarithmic concentrations of samples (μg protein or μM HbO2), a straight line relationship was observed. The ranges of kidney protein concentration in above relation are 62.52000μg in Altschule and Levis' method, 25400μg in Maren's method and 6.25100μg in Wilbur and Anderson's method. The ranges of liver protein concentration in above dose-response relation is slightly higher than that in the kidney. The ranges of blood hemoglobin (μM) in straight line relationship are 0.258μM in manometric method, 0.01951.25μM in Maren's method and 0.00780.125μM in Wilbur and Anderson's method.2. In above three methods, Wilbur and Anderson's method was most sensitive for measuring the enzyme activity.3. For the preparation of kidney and liver samples, the well-washing of the tissue pieses may be sufficient to omitt the blood contamination, because the enzyme activity obtained from well-washed organ is similar to the enzyme activity obtained from perfused organ.4. Inhibition grade of carbonic anhydrase activity by acetazolamide administration (20 mg/kg, i.p., 2 hr value) was examined. Kidney and blood carbonic anhydrase activities were inhibited by acetazolamide. Its inhibition grade is different by assay method and the grade is similar in both Altschule and Levis' and Maren's methods, however, the inhibition was appeared more strongly by Wilbur and Anderson's method.5. Liver carbonic anhydrase activity was not inhibited by acetazolamide in above three methods.

Effects of aldosterone and puromycin, alone or in combination, on kidney carbonic anhydrase and ATPase activities and/or urinary excretion of sodium were investigated using adrenalectomized and normal mice to clarify the significance of above enzymes in induction hypothesis of aldosterone.1. In adrenalectomized mice, puromycin had no effect on the activities of kidney carbonic anhydrase and Na+-K+-ATPase and on urinary excretion of 22Na.2. Combined administration of puromycin with aldosterone inhibited both the elevation of kidney carbonic anhydrase activity and decrease of urinary excretion of 22Na induced by aldosterone alone, whereas Na+-K+-ATPase activity had no relation with puromycin.3. Time-course of effect of puromycin was examined in normal mice. Puromycin (1-125 mg/kg, i.p.) inhibited kidney carbonic anhydrase activity and its maximum effect was observed at 4 hr after injection without any alterations in Na+-K+-ATPase activity.In conclusion, the elevation of kidney carbonic anhydrase activity by aldosterone in adrenalectomized mice may be due to the increased synthesis of this enzyme, which may have a correlation with aldosterone-induced decrease of urinary excretion of sodium.

Time-course of effect of aldosterone on the incorporation of 14C-orotic acid into kidney RNA from adrenalectomized and normal mice was examined with the following results.1. Stimulatory effect of aldosterone (0.55 μg/kg, s.c.) on 14C-orotic acid incorporation into RNA from kidney homogenate and its nuclear fraction was observed in early stages (1.53 hr) after aldosterone administration in adrenalectomized mice, however, the incorporation was not changed at 5 hr after aldosterone. RNA content in homogenate and subcellular fractions did not change in any cases.2. After the administration of larger dose of aldosterone (50μg/kg) to adrenalectomized mice, 14C-orotic acid incorporation into kidney RNA was not changed until 3 hr.3. Cortisol had no effect on RNA specific radioactivity and RNA amount in adrenalectomized mouse kidney. 4. 14C-orotic acid incorporation into kidney RNA was not changed until 5 hr between normal and adrenalectomized mice.5. 14C-orotic acid incorporation into kidney nuclear RNA from normal mice was increased until 3 hr after aldosterone (2 and 20μg/kg) administration and the maximum effect was observed at 1.5 hr later.6. The early effects of aldosterone on RNA synthesis in adrenalectomized mouse kidney are agreed with the general concept in “induction hypothesis” which has been offered by the study using adrenalectomized rat kidney and/or urinary bladder of toad.

With increased application of radioactive pharmaceuticals, exposure of patients to radiation has attracted grave attention. Especially when mercuric Preparations are used, the kidney is exposed to a large dosis. To eliminate the radioactive substance as early as possible after receiving diagnostic informations is one of the measures to relieve the patient from radiation injury.Male dd N mice were used as experimental animals. Tracer doses of various radioactive mercuric preparations were intraperitoneally injected to them, and radioactivity was daily measured for 3 days with animal bodies and their excrements, using the whole-body animal counter, and at 3 days radioactivity of 203Hg in various organs was determined with the well-type scintillation counter. From immediately after the radioisotope injection, various drugs (in doses varying by toxicity) were given every 24 hours, that is, once daily for 3 days and effects on the retention, excretion and organ distribution of 203Hg were examined to evaluate the eliminating activities.Drugs that decreased the retention of 203Hg-MHP and accelerated its urinary excretion were BAL, mercaptoacetic acid (MAA), 2-mercaptopropionyl glycine (MPG), DL-penicillamine (Pen.) and GSH. Furosemide conversely increased the retention. L-CySH, Rongalite and EDTA elicited no change.Drugs that were effective for elimination of 203Hg chlormerodrin were MPG, MAA, and Pen. BAL lowered 203Hg concentration in the kidney, but elevated it in the liver and brain. GSH decreased 203Hg in the brain and pancreas, but did not change its amount in the other organs. Furosemide increased 203Hg in the liver and pancreas. EDTA, DTPA and Rongalite exerted no effect.Drugs that were effective for eliminatin of 203Hg-Salyrgan were MPG, GSH, Pen and Rongalite. Furosemide and BAL increased the retention.All the mercuric compounds were increased in body retention under fasting condition. Effect of eliminating them differed in different compounds.The distribution and excretion of these mercuric compounds administered s.c. or i.p. into Ehrlich ascites cancer-bearing mice were also examined.

203HgCl2 was given to mice, and its retention, excretion and distribution were investigated, Furthermore various drugs were examined for 203 Hg-expelling effect.1. Retention : The retention after the s.c., i.p. and p.o. administration were daily explored for 3 days. The retention after s.c. administration was highest, and next in order were i.p. and p.o. administration.2. Distribution : The distribution of 203 HgCl2 was determined times daily for 3 days after i.p. administration. The radioactivity was highest in the kidney already at 1 hour, and attained the peak at 4 hours.The subcellular distribution of 203 HgCl2 in the liver and kidney at 24 hours after i.p. administration was greater in the supernatant fraction from both organs than in the other fractions.3. Effects of administration of various drugs : 3-day experiments on the whole body : The retention of 203 HgCl2 after its i.p. administration was decreased by BAL and D-Penicillamine. 203 Hg Activity was lowered in the kidney, liver, pancreas, spleen, brain and blood by BAL, in the kidney, pancreas and spleen by D-Penicillamine, in the brain by L-CySH·Gly, in the brain and blood by L-CySH, in the kidney by 2-Mercatopropiony glycine and also by Pyridonine-4SH, in the spleen, brain and blood by Mercaptoacetic acid (though it increased 203 Hg concentration in the kidney), and in the kidney be EDTA and DTPA (though increased in the pancreas and brain).4. Effect of starvation : The fecal excretion of 203 HgCl2was decrease and its retention was increased by the starvation. The absorption of 203 HgCl2 was increased by the starvation.

Various drugs (anti-cancer agents, chelating agents, amino acids and salts) were injected i.p. with 65Zn into Ehrlich ascites cancer-bearing mice, and after 1 hr its effect on the organ distribution of 65Zn was investigated.1. Mitomycin C (200 μgg/mouse), Toyomycin (50 μg/mouse) and Nitromin (500 μg/mouse) had no effects on the distribution of 65Zn. Merphyrin (500 μg/mouse) caused a decrease in 65Zn uptake by cancer cells without any effects in the other organs.2. EDTA and BAL (each 1 mg/mouse) inhibited 65Zn> uptake by cancer cells and conversely elevated its concentration in ascites supernatant. BAL elevated also 65Zn concentration in other various organs. Tetracycline, dihydrothioctic acid and Na-thioglycolic acid (each 1 mg/mouse) did not affect the 65Zn uptake by cancer cells.3. Penicillamine (D-and DL-type, each 1 mg/mouse) caused a decrease of 65Zn concentration in cancer cells with an increase of its concentration in the other organs. Cysteine, histidine and glycine (each 1 mg/mouse) had no effects on the 65Zn distribution in every organs.4. Various salts (each 0.1 ml of 3.5 ×10-2M solution was injected per mouse) also affected organ distribution of 65Zn. ZnSO4, CoCl2 and NiSO4 inhibited the uptake of 65Zn by cancer cells, however, MgCl2, MnSO4, FeCl

Organ distribution of 65Zn injected i.p. or s.c. into Ehrlich ascites cancer-bearing mice was examined with the following results.1) Distribution of 65Zn injected i.p. was examined as a function of time (1-24 hr). 65Zn was found mainly in ascites cancer cells reaching a maximum (26%) 6 hr after the injection and its concentration was gradually decreased thereafter. Radioactivity of 65Zn in liver, kidney, pancreas and spleen were less than that in cancer cells. Disappearance of the radioactivity contained in cancer cells was very little after washing.2) Subcellular distribution pattern of 65Zn in ascites cancer cells was examined extending from 1 to 24 hr after i.p. injection. The amount of 65Zn in supernatant fraction was highest followed by nuclear, microsomal and mitochondrial fractions in the descending order and the above distribution pattern was same at any times.3) Radioactivity of 65Zn administered s.c. into cancer-bearing mice was significantly increased in the liver and decreased in pancreas and blood with no changes in the kidney and spleen compared with that in normal mice. After the i.p. injection of 65Zn into cancer-bearing mice, 66% of total radioactivity was found in ascites and 65Zn concentration in the liver, kidney, pancreas, spleen and blood were less than that in normal mice.4) When the radioactivity of 65Zn injected i.p. or s.c. in the subcellular fractions of liver and cancer cells are expressed per mg protein, the highest concentration of 65Zn in the liver was found in the supernatant fraction followed by microsomal, nuclear and mitochondrial fractions in the decending order. These distribution pattern of 65Zn was equal with that of cancer cells.5) 65Zn in the liver and cancer cells binds loosely with protein. After the treatment of each subcellular fractions obtained from liver and cancer cells with (NH4) 2SO4 (70% saturation) and trichloroacetic acid (10% in final), most of 65Zn was distributed in supernatant excepting the liver nuclear fraction in which most of radioactivity exists in precipitate by (NH4) 2SO4.6) Subcellular distribution of 65Zn in mammary carcinoma was examined 24 hr after the s. c. injection. 65Zn concentration is equal in nuclear and supernatnat fractions followed by microsomal and mitochondrial fractions in the descending order. The distribution pattern of 65Zn in nuclear fraction was different from that in the liver.

The distribution and excretion of 60CoCl2 administered subcutaneously or intraperitoneally into Ehrlich ascites cancer-bearing mice were examined with the following results.1. The concentration of 60Co in various organs of cancer-bearing mice was generally a compared with the control, especially in kidney and blood. The urinary and fecal excretion of 60Co in 24 hours had no differnce in control and experimental groups. The uptake of 60Co by ascites cancer cells was more compared with the other organs.2. In ascites cancer intraperitoneally implanted mice, the time course of the changes of 60Co distribution were examined. At 6 hours after the intraperitoneal injection, 60Co concentration in ascites cancer cells was highest, but later its concentration decreased gradually.3. At 1, 6, 12 and 24 hours after the intraperitoneal administration of 60Co, its distributionin subcellular fractions of Ehrlich ascites cancer cells was examined. Nuclear fraction had the highest concentration, followed by supernatant, microsomal and mitochondrial fractions in the descending order, and these concentrations did not relate to the lapse of hours after the 60Co administration.4. Concerning the effects of continual administration of anticancer agents to cancer bearing-mice, Mitomycin C and hydrocortisone acetate inhibited the ascites retention and corrected the disturbances of 60Co distribution observed in kidney and blood. Toyomycin inhibited the ascites retention without any effects on 60Co distribution. Nitromin, Tespamin and Merphyrin had no effects on grade of ascites retention and concentration of 60Co in several organs compared with the control.5. Soon after the intraperitoneal administration of 60Co to cancer intraperitoneally implanted mice, anticancer agents, amino acids, chelating agents and various salts were administered intraperitoneally and after 1 hour their effects on the distribution of 60Co were examined, On the effects of anticancer agents, Toyomycin inhibited 60Co uptake by cancer cells, but Nitromin increased it conversely, Mitomycin, Tespamin and Merphyrin did not affect 60Co uptake by cancer cells, In any cases the distribution of 60Co in other organs did not change compared with the control, On the effects of amino acids, Penicillamine, Cysteine and and Histidine inhibited 60Co uptake by cancer cells, but Glycine had no effect on it. In many cases 60Co concentration in the kidney was decreased, On the effects of chelating agents, EDTA, BAL and Na mercaptoacetate inhibited 60Co uptake by cancer cells, but dihydrothioctic acid increased it conversely, 60Co distributions in other organs were varied in case and case, On the effects of salts, Co2+, Mn2+, and Zn2+ inhibited 60Co uptake by cancer cells and increased 60Co retention in ascites supernatant, Mg2+, Fe2+, Ni2+ and Cu2+ did not affect both 60Co uptake in cancer cells and 60Co retention in ascites supernatant. 60Co distributions in other organs were varied in case and case and clear definite tendency did not observe in the 60Co distribution among the organs.

In the first, Na22Cl, Rb86Cl and Cs137Cl were subcutaneously given in a dose of 1μc/ 0. 1cc to male mice of dd strain weighing 2025g following right adrenalectomy and left adrenal enucleation (UAE) or bilateral adrenalectomy, and the excretions and distribution of each in the body was investigated and regenerating adrenal glands were histologically studied. Next, using rats weighing about 200g serum Na and K concentration following UEA and effects of various hormones upon it were investigated with the following results.1) Within 1 week after UAE, Na22 excretion rate was in inverse relation with those of Rb86 and Cs137, but all returned to the, normal after 1 week. Within 1 week the concentrations of Rb86 and Cs137 in various organs were significantly increased but returned to normal after 2 weeks.2) Histologically, regeneration of mouse adrenal gland observed at 4th day after UAE and regenerating cell was generally presented fascicular arrangement. Adrenal glands were restored to normal almost at 18th day after UAE, but zonal formation is not clear. At 35th day, its state is like to adrenal medullectomy.3) Serum Na and K concentration were returned to normal within 12 weeks after UAE, and it was not related to zonal formation.4) As to the influences of hormane administration to serum Na and K concentration, definit relation was not gained on adrenal corticoids and ACTH administration, administration of adrenalin and growth hormone produced no remarkable change, DOCA administration increased serum Na concentration.From the results of present experiments, histological finding is not keep with the restoriction of mineral metabolism. There is a scope to criticized to so-called zonal theory, that is, early regenerating adrenal gland cells after UAE have mineral and gluco metabolism functions.

203Hg-ethylmercury chloride was orally given to mice and rats, and its absorption, retention, excretion and distribution were investigated.1. Absorption : The absorption was considerably fast after the oral administration as in the case of methylmercury.2. Retention in the body : The daily decrement curve of the whole-body retention represents the sum of two exponential functions in rats, but a single expornential function in mice.3. Distribution in the body : The distribution of 203Hg was determined for 3 days with mice and 7 days with rats after the oral administration. In rats, the highest radioactivity was found in the kidney followed by in blood, liver, spleen, pancreas and brain in the descending order. In mice, the highest radioactivity was also found in the kidney, however, next order was the liver, pancreas, spleen, blood and brain successively.4. Excretion : The daily excretion of 203Hg was high in feces and low in urine.5. Effects of administration of various drugs. In 3 day experiments with mice, the effective drugs on eliminating of 203Hg are DL-Penicillamine, 2-mercaptopropionyl glycine and GSH. BAL, Mercaptoacetic acid and Thioethanol were found to increase 203Hg concentration in the brain. In 7 day experiments with rats, the effective drugs in expelling 203Hg were BAL and 2-Mercaptopropionyl glycine.