Status of the toxicity reports of pesticides for their registration in Finland.

Recently, attention has centered on the validity of toxicity reports for the registration of pesticides. In Finland, a study was conducted to evaluate the validity of the toxicity reports of pesticides done in U. S. laboratories. Twenty-seven pesticide formulations of the 230 on the market in Finland were selected for the study. The material contained 61 toxicity reports of 16 pesticide formulations done in U. S. laboratories considered questionable, 64 reports from U. S. laboratories considered reliable, and 142 reports originating from European laboratories, totally 267 reports. These reports were divided into valid according to the Organization for Economic Cooperation and Development (OECD) guidelines, and included those with minor discrepancies but in compliance with the good laboratory practice at the time of the study (group I), and and invalid according to the OECD guidelines (group II). After a closer evaluation of the 125 U. S. reports, 85 reports were considered valid and 40 invalid. As a function of time, the proportion of valid studies of all studies on pesticides for the registration purposes in Finland increased during 1961-1981 from 47 to 93%, being 100% at the present time. The lack of information of pesticide formulations registered earlier creates a problem to be solved in the future in Finland, rather than the lack of the validity of toxicity reports.

Influence of environmental temperature on the induction of xenobiotic metabolism by beta-naphthoflavone in rainbow trout, Salmo gairdneri.

The response of hepatic microsomal cytochrome P-450 monooxygenase and UDPglucuronosyltransferase to a common inducer, beta-naphthoflavone, was investigated in rainbow trout acclimated to 5 or 17 degrees C. The hepatic microsomal 7-ethoxycoumarin-O-deethylase, 7-ethoxyresorufin-O-deethylase, and benzo[a]pyrene hydroxylase activities, measured at 5 or 17 degrees C, were induced at both acclimation temperatures. Maximum response of 7-ethoxyresorufin-O-deethylase and 7-ethoxycoumarin-O-deethylase activities to beta-naphthoflavone was obtained with doses of 5 mg/kg and above, both in warm- and cold-acclimated trout. In warm-acclimated trout the maximum degree of induction of cytochrome P-450-dependent activities was already evident on the day following treatment with beta-naphthoflavone (100 mg/kg), whereas these activities reached maximum values after 3 days in cold-acclimated fish. Furthermore, the maximum induced values of 7-ethoxyresorufin-O-deethylase and benzo[a]pyrene hydroxylase activities were higher in cold-acclimated than in warm-acclimated fish. beta-Naphthoflavone treatment caused a slight but significant increase (1.6-fold) in UDPglucuronosyltransferase activity in warm-acclimated fish when measured at 5 or 17 degrees C, whereas in cold-acclimated fish a significant elevation was seen only when the activity was determined at environmental temperature (5 degrees C).

Catalytic and immunological comparison of coumarin 7-hydroxylation in different species.

Coumarin 7-hydroxylation activities were determined in the liver microsomes of eight different species: pig, mouse (three strains), rat, hamster, guinea-pig, rabbit, rainbow trout and crayfish. A high coumarin 7-hydroxylase activity was found in the microsomes of D2 mouse, rabbit, hamster and pig, an intermediate activity in case of B6 and AKR mice, rat and guinea-pig and a very low activity in rainbow trout and crayfish. In Ouchterlony immunodiffusion analysis our antibody against coumarin 7-hydroxylase specific cytochrome P-450 from D2 mice was able to form a weak precipitin line only with rat and hamster microsomes in addition to the three strains of mice (D2, B6, AKR) where the band was very sharp. Strongest inhibition by the antibody (50% or more) of microsomal coumarin 7-hydroxylase was found in the case of the three strains of mice and rabbit. In the case of the other species the inhibition was very weak or did not exist.

Xenobiotic biotransformation in the rainbow trout liver and kidney during starvation.

Microsomal cytochrome P-450-dependent activities in the kidney of fish starved for 6 weeks were significantly lower than in fed fish whereas these activities in the liver were only depressed after 12 weeks of starvation. Hepatic cytochrome P-450-dependent activities were depressed to varying extents after 12 weeks of starvation when different substrates were used. The content of hepatic cytochrome P-450 was not affected by starvation. Hepatic UDP-glucuronosyl transferase activities were not affected by starvation. Induction of several hepatic cytochrome P-450-dependent activities by treatment of fish with beta-naphthoflavone was not influenced by starvation. In the kidneys of fish starved for 12 weeks induced levels of cytochrome P-450-dependent benzo(a)pyrene hydroxylase activities were significantly lower than in the kidneys of fed induced fish.

Influence of fly ash and charcoal on the inducer activity of benzo[a]pyrene and smoke condensate in the gut mucosa of the rat.

Fly ash from either peat- or coal-fired power plants is unable to block the inducer activity of benzo[a]pyrene in the gastrointestinal tract. Activated charcoal retains adsorbed polycyclic aromatic hydrocarbons during passage through the gut so that no induction of polysubstrate monooxygenase occurs in the mucosa.

Thermal acclimatization of hepatic polysubstrate monooxygenase and UDP-glucuronosyltransferase of mature rainbow trout (Salmo gairdneri).

The thermal acclimatization of hepatic cytochrome P-450-dependent polysubstrate monooxygenase (PSMO) and UDP-glucuronosyltransferase of mature rainbow trout (Salmo gairdneri) was studied. The results indicate that the PSMO system, 7-ethoxycoumarin and benzo(a)pyrene as substrates, shows almost ideal acclimatization pattern in autumn during water cooling. The enzyme activities were identical if measurements were carried out at acclimatization (=environmental) temperatures which were 20 degrees C in August and 5 degrees C in November. If a constant incubation temperature (18 degrees C) was used, the PSMO activities were significantly higher in cold (5 degrees C)-acclimatized fish. The acclimatization process could be seen both in specific and total activities. The content of cytochrome P-450 remains at constant level from August to November. In early summer during water warming the PSMO activities increased considerably in both sexes in all incubation conditions. The specific and total UDP-glucuronosyltransferase activities were significantly higher in warm-acclimatized fish both in the autumn and in the spring if the activities were measured at environmental temperature. No differences could be detected if the measurements were carried out at constant experimental temperature (18 degrees C).

Increased fluidity of a model membrane caused by tetrahydro-beta-carbolines.

Alterations in membrane fluidity caused by alcohols and tetrahydro-beta-carbolines (THBCs) have been studied. Dipalmitoylphosphatidylcholine vesicles were used as a membrane preparation, and changes in the fluidity were revealed by two fluorescent probes: 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS) and N-phenylnaphthylamine (NPN). It was found that THBCs, which are condensation products of tryptamine and formaldehyde or acetaldehyde, were at least 2 orders of magnitude more potent in causing fluidity changes than the comparable alcohols (methanol and ethanol). Both 1,8-ANS (binding close to the polar end of the phospholipid molecules) and NPN (binding to the hydrophobic region of the membrane) were able to reveal changes in membrane fluidity, although there were differences between the behavior of the two probes. The condensation product of acetaldehyde--the primary metabolite of ethanol--and tryptamine were found to be 200-300 times more potent in causing fluidity changes than ethanol itself (as determined with both 1,8-ANS and NPN).

Tissue concentrations and interaction of zinc with lead toxicity in rabbits.

Adult male rabbits were exposed to lead (0.2%), zinc (0.5%) or to both lead and zinc (0.2% and 0.5%) which were given in the drinking water as acetates for 2 weeks or 4 weeks. Blood lead levels in lead exposed rabbits were about 15-fold in comparison with the controls after 2 weeks exposure and remained at this level after further exposure, but when zinc was given with lead the blood lead level doubled from 2 weeks to 4 weeks. In rabbits having both lead and zinc, the cerebral lead concentration was much lower than in rabbits exposed only to the respective amount of lead. No such effect was found in the cerebellum or sciatic nerve. Zinc did not prevent lead accumulation in the parenchymal tissues. Only a rather low induction of drug metabolizing enzyme activities was found in the liver of lead-exposed rabbits and zinc did not modify this induction. The results suggest that, although zinc might delay the lead accumulation in the cerebrum, it has little value in preventing peripheral neuropathy or metabolic alterations caused by lead.

Structural and biotransformational membrane changes in the liver and intestine during chronic ethanol administration.

The binding of a fluorescent probe 1-anilinonaphthalene-8-sulphonic acid (1,8-ANS) to liver microsomal membranes was markedly increased after chronic ethanol administration while the binding of a non-ionised probe phenylnaphthylamine (PNA) was not altered. The increase in 1,8-ANS binding is in accordance with the simultaneous increase of the ethoxycoumarin O-de-ethylase activity and cytochrome P-450 concentration. Also the intestinal ethyoxycoumarin O-de-ethylase activity and cytochrome P-450 concentration were increased. No changes in the aryl hydrocarbon hydroxylase or UDP-glucuronosyltransferase activities were found. The chronic ehtanol administration increased the phospholipid amount in the liver microsomes and altered the fatty acid composition of microsomal phospholipids by decreasing the amount of oleic acid and increasing linoleic acid proportion. The data suggest that chronic ethanol administration may effect the biotransformation enzyme activities by changing the structural properties of the membranes as well as increasing the cytochrome P-450 concentration.

Differences in the response of hepatic and intestinal drug metabolizing enzymes in rats following carbon tetrachloride and/or phenobarbital treatment.

The activities of the microsomal drug metabolizing enzymes in the liver and intestinal mucosa of rats were studied after the intraperitoneal administration of carbon tetrachloride and/or subcutaneous phenobarbital administration. The membrane phospholipid content was decreased after carbon tetrachloride treatment indicating destruction in the membrane structure. Aryl hydrocarbon hydroxylase activity was decreased in the liver and intestinal mucosa after treatment with CCl4 alone and in combination with phenobarbital. The CCl4 treatment increased the intestinal epoxide hydratase activity but decreased the activity in the liver. The hepatic UDPglucuronosyltransferase was slightly induced by phenobarbital and the activity was elevated by the CCl4 treatment. In the intestinal mucosa the enhanced UDPglucuronosyltransferase activity was observed only after phenobarbital pretreatment and the activity was decreased by CCl4. These results support the view that epoxide hydratase and UDPglucuronosyltransferase enzymes occupy different locations in the endosplasmic reticulum of intestinal mucosa than of liver.

Effect of administration route of 3-methylcholanthrene on the inducibility of intestinal drug-metabolizing enzymes.

The inducibility of the mucosal drug-metabolizing enzymes of rat small intestine was studied by administering 3-methylcholanthrene either intragastrically or intraperitoneally. The aryl hydrocarbon hydroxylase activity was 4.1 times higher after intragastric than after intraperitoneal administration of methylcholanthrene. The ethoxycoumarin-O-diethylase activity was 11 times and UDP-glucuronosyltransferase (with p-nitrophenol as substrate) activity was 3 times higher after intragastric administration than after intraperitoneal administration. The epoxide hydratase activity was, on the other hand, 38% lower after intragastric administration of 3-methylcholanthrene than after intraperitoneal administration. The results suggest that compounds entering the body intragastrically, e.e. in the diet, might have profound enzyme-specific effects on the intestinal metabolic rates of drugs.

Hepatic drug metabolism during ethanol ingestion in riboflavin deficient rats.

Riboflavin deficiency was induced by feeding rats a riboflavin-deficient diet for 1 month. In order to find out if there are any combined effects of ethanol and riboflavin deficiency on drug metabolism, a group of riboflavin-deficient rats were also given ethanol in their drinking water. At the end of the feeding period, hepatic drug-metabolizing enzyme activities were determined. The hepatic phospholipid and protein contents were the same in rats receiving a standard diet and in those on a riboflavin-deficient diet. However, ethanol ingestion in both groups enhanced significantly the phospholipid content. Ethanol ingestion also markedly enhanced the hepatic cytochrome P-450 concentration in rats fed either a standard or riboflavin-deficient diet. Ethoxycoumarin O-deethylase activity was significantly lower in riboflavin-deficient rat livers than in those of the controls. In both groups ethanol ingestion nearly doubled the activities. Aryl hydrocarbon hydroxylase activity was also significantly decreased during riboflavin deficiency. However, ethanol administration did not change the activities of this enzyme. UDP glucuronosyltransferase activity was slightly lower in riboflavin-deficient rat livers than in those fed a standard diet. No significant decrease was found in the epoxide hydrase activity in the riboflavin-deficient rats. However, the riboflavin-deficient rats had enhanced activity after the ethanol ingestion.