In order to establish sensitive methods of detecting minor renal damage, changes of enzymes, tubular cell counts, and creatinine in the urine were investigated in rats that had been given nephrotoxic chemicals. Daily administration of mercuric chroride (HgCl2) dose-dependently increased urinary excretions of lactate dehydrogenase (LDH), aspartate aminotransferase (GOT), alkaline phosphtase (ALP), leucine aminopeptidase (LAP), lysozyme (LZM), N-acetyl-β-D-glucosaminidase (NAG), and .acid protease together with increased counts of tubular cells in the urine. The increase in tubular cell counts and the change in urinary LDH isoenzyme profile preceded the changes in the other enzymes. Daily administration of gentamicin (GM) increased urinary excretions of LDH, GOT, LZM, NAG, acid protease and tubular cell counts in a dose-dependent manner, but did not increase γ-glutamyl transpeptidase (γ-GTP) and ALP excretions. The urinary isoenzyme profiles of LDH in rats treated with GM were different from those with HgCl2. The increase in acid protease excretion outlasted those in LDH and GOT in the high dose group. It was concluded that the severity of renal damage can be readily detected by periodic determinations of the following urinary parameters : tubular cell counts, LDH isoenzyme, acid protease, LZM and NAG, in addition to either LDH or GOT and one of the enzymes ALP, LAP or γ-GTP. Furthermore, the site of renal damage can be presumed from these results.
In order to establish sensitive methods of detecting minor renal damage, changes of enzymes, protein, tubular cell counts, and creatinine in the urine were investigated in rats to which nephrotoxic chemicals had been administered. Daily administration of p-aminophenol (PAP) dose-dependently increased urinary excretions of lactate dehydrogenase (LDH) and its isoenzymes (LDH5=LDH4>LDH3>LDH2=LDH1), aspartate aminotrans-ferase (GOT), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GTP), leucine aminopeptidase (LAP), lysozyme (LZM), N-acetyl-β-D-glucosaminidase (NAG) and acid protease together with increased counts of tubular cells in the urine. Tubular cell counts, LDH and GOT were more sensitive indicators in the PAP tubulonephritis. Single i.v. injection of puromycin aminonucleoside (PM) dose-dependently increased urinary excretions of LDH and its isoenzymes (LDH1=LDH5>LDH2=LDH4>LDH3), GOT, NAG, acid protease and protein but degree of the increases in these enzymes was lower than those in the rats treated with PAP. PM increased excretions of high molecular weight proteins but did not increase ALP, γ-GTP, LAP, LZM and tubular cells excretions. Single i.v. injection of hexadimethrine increased urinary excretion of LDH and its isoenzymes (LDH1=LDH5>LDH2>LDH3=LDH4), GOT, LZM, NAG and acid protease together with increased counts of tubular cells in the urine but did not increase ALP, γ-GTP and LAP excretions. It is concluded that tubular cell counts, LDH isoenzymes and battery of these enzymes in urine are useful markers for detecting the severity and the site of renal damage in addition that urinary protein is a useful marker for detecting glomerular damage.
A personal computer was introduced in conditioned lever-press avoidance experiment in Sidman situation. Experimental data were analyzed with both classic indices such as response rate and shock rate and new indices introduced in this experiment. New indices include sum of squares of shock-shock intervals, mean of shock-response time and number of shock-response time less then 5 sec. Results obtained in diazepamtreated mice were consistent with those reported previously by the classic method. In mannitol-treated mice, significant changes of the avoidance behavior could be detected by both the classic index and our new indices. By the new indices other than classic indices, the effect of administration with sodium chloride on the avoidance behavior could be detected, whereas the classic indices could not detect the effect. Though the meanings of these indices in physiological and toxicological aspects should be appreciated in future study, the experimental system reported in this paper might be applicable in the toxicological research to detect the effect of administration with various substances on the avoidance behavior.
Chinese hamster B241 cells were treated with 5 nM allylisothiocyanate (AI) or 10 nM trans-cinnamaldehyde (CA) and surviving cells were cultivated for generations until the cells acquired the characteristics of transformed cells based on in vitro criteria : increase in (a) saturation density in monolayer culture, (b) plating efficiency at low serum level and (c) colony forming efficiency (CFE) in soft agar medium. When the values of CFE of the treated cells had become significantly high, anchorage-independent colonies were isolated, propagated and then subjected to an assay for neoplastic transformation. The anchorage-independent clonal cells (CH-AI-AG+-1 and CH-CA-AG+-2) were subcutaneously injected into a suprascapular site in nude mice. The mice were maintained in an SPF animal care facility and observed for tumor formation. Growth of neoplasm at the injection sites was observed in 6 out of 7 mice and in all of the 6 mice injected with CA- and AI-transformed cells, respectively, during 3 to 8 months after the injection, as compared with 1 out of 6 mice injected with untreated control cells. Subsequent transplantation of the tumor cells into new mice induced tumor production at the injection site in all the animals within a considerably shorter period of time than that following the initial inoculation. Malignancy of the neoplastic cells was ascertained by histological examination, and the cells were found to have karyotypes of the hamster cells after in vitro cultivation of the tumor cells. These experimental results suggest the transforming potency of the flavoring agents in Chinese hamster cells.
d·d-T80-prallethrin, a pyrethroid insecticide for sanitary use, was administered to Crj : CD (Sprague Dawley) rats at concentrations of 120, 600 or 3, 000 ppm in diet for one year to assess the chronic toxicity potential and the reversibility. The summarized results obtained are as follows : 1. Chronic toxicity study 3, 000 ppm : Decreases in body weight gain, food consumption, and water intake were observed. Slight alopecia in the neck and/or back was noticed during the first and second weeks, but the animals were recovered thereafter. Slight anemic changes such as decreases in hemoglobin concentration, hematocrit value, MCV and MCH were observed in the females3 at 52 week. Blood biochemistry revealed increases in total cholesterol (in the males and females at 13, 26 and 52 weeks), phospholipid (in the males and females at 13, 26 and 52 weeks), albumin (in the males at 13 and 26 weeks, in the females at 52 week), total protein (in the males at 26 week, in the females at 52 week), A/G ratio (in the males at 13 week, in the females at 26 week), creatinine (in the males at 52 week), urea nitrogen (in the females at 52 week), GOT (in the males and females at 52 week) and GPT (in the males and females at 52 week), and decreases in triglyceride (in the females at 26 and 52 weeks) and alkaline phosphatase (in the males at 13 and 52 weeks). In urinalysis, an increase in bilirubin was observed in the males at 52 week. Gross-pathology revealed a lower incidence of accentuated lobular pattern of liver (in the males at 26 week) and a higher incidence of enlarged liver (in the males at 52 week). In organ weight, increases in liver (in the males and females at 26 and 52 weeks), kidney (in the males at 26 and 52 weeks) and thyroid weights (in the males at 26 and 52 weeks, in the females at 26 week), and decreases in spleen (in the females at 26 and 52 weeks) and adrenal weights (in the females at 52 week) were observed. Histopathological examination revealed a lower incidence of fatty metamorphosis in the liver of females at 52 week. 600 ppm : An increase in liver weight was observed in the males at 26 week. 120 ppm : No effect was observed. 2. Reversibility study Almost all the above chronic toxicities were reversible. However, blood biochemistry revealed increases in albumin (in the females), A/G ratio (in the females) and creatinine (in the males) and a decrease in alkaline phosphatase (in the males) at 3, 000 ppm. Based on the above results, one year d·d-T80-prallethrin administration revealed slight toxic effects mainly in the liver, kidney and thyroid at the dose level of showing body weight gain suppresion. However, most of the toxic effects were reversible after 13-week withdrawal period. It is concluded that no-observed effect level is 600 ppm (43.3 mg/kg/day) for female and 120 ppm (7.16 mg/kg/day) for male.