A comparison of metal levels and antioxidant enzymes in freshwater snails, Lymnaea natalensis, exposed to sediment and water collected from Wright Dam and Lower Mguza Dam, Bulawayo, Zimbabwe.

We compared the bioaccumulation of lead (Pb), cadmium (Cd), zinc (Zn), copper (Cu), nickel (Ni) and iron (Fe) with antioxidant enzyme activity in tissues of the snails, Lymnaea natalensis, exposed to elements of two differently polluted dams. 45 snails were exposed to sediment and water collected from Wight Dam (reference) whilst another 45 snails were also exposed to sediment and water collected from Lower Mguza Dam (polluted dam). Except for Fe in sediment and Pb in water, metal concentrations were statistically higher in sediment and water collected from Lower Mguza Dam. Lead, Cd and Zn were two times higher in tissues of snails exposed to Lower Mguza Dam elements. On one hand, superoxide dismutase (SOD), diphosphotriphosphodiaphorase (DTD) and catalase (CAT) activities were significantly lower whilst malondialdehyde (MDA) levels were significantly higher in tissues of snails exposed to Lower Mguza Dam sediment and water. On the other hand, selenium-dependent glutathione peroxidase (Se-GPX) activity was significantly elevated in tissues of snails exposed to Lower Mguza Dam sediment and water. Snails exposed to Lower Mguza Dam elements seem to have responded to pollution by increasing CAT and Se-GPX specific activity in an effort to detoxify peroxides produced as a result of metal induced oxidative stress.

Metal accumulation and antioxidant enzyme activity in C. gariepinus, catfish, and O. mossambicus, tilapia, collected from Lower Mguza and Wright Dams, Zimbabwe.

The aim of this study was to measure antioxidant enzyme activities as biological indicators of pollution in tissues of two species of fish. Five Clarius gariepinus and three Oreochromis mossambicus were collected from Umguza Dam (polluted dam) whilst seven C. gariepinus and eight O. mossambicus were collected from Wright Dam (relatively pristine dam). Diphosphotriphoshodiaphorase and catalase activities were consistently lower (42 +/- 2% and 78 +/- 20%, respectively) in liver whilst malondialdehyde levels were two times higher in muscles of both species of fish collected from Umguza Dam. However, seleniumdependent glutathione peroxidase (Se-GPX) activity was elevated four-fold in liver and gills of O. mossambicus collected from Umguza Dam. Metal levels were two to five times higher in muscles of both species of fish collected from Umguza Dam. Fish from Umguza Dam seem to have responded to pollution by increasing Se-GPX specificactivity in an effort to detoxify peroxides produced as a result of metal induced oxidative stress.

The influence of dietary fat on hepatic bioactivation of aflatoxin B1 in rats.

Fischer 344 rats were fed a low-fat high carbohydrate (HC) diet, an isocaloric fat-containing (IC) diet, a hypercaloric fat-containing (HF) diet or a commercial rodent chow. The effects of these diets were studied on the binding of aflatoxin B (AFB1) to exogenous DNA, and on the activities of hepatic glutathione transferases (GSTs), cytochromes 2B1 and 1A1. Microsome-mediated binding of [3H]AFB1 to exogenous DNA was significantly lower in the HC-rats than in the chow and IC-fed rats. No significant differences were noted between HF and either HC or IC rats. There was no significant difference in hepatic GST activity of rats fed the different diets. Our results suggest that high-carbohydrate low-fat diets reduce microsome mediated epoxidation of AFB1 to a larger extent than high-fat diets. In general, high fat diets increased cytochrome 1A1 and 2B1 activities relative to chow and high carbohydrate diet. This suggests greater detoxification of AFB1, thus reducing the amount of AFB1 available for hepatic macromolecular binding.

The effect of diet on aflatoxin B1 binding to hepatic macromolecules in rats.

Fischer 344 rats were fed a low-fat high carbohydrate diet (HC), an isocaloric fat-containing diet (IC), a hypercaloric fat-containing diet (HF) or rat chow. Covalent binding of AFB1 to liver DNA, RNA and total proteins was investigated in a 24 hour period following administration of a single intraperitoneal dose of AFB1 (1 mg/kg body weight). AFB1 binding to nucleic acids was greatest in the HC and was generally significantly lower (p < 0.05) in the HF, IC and rats fed chow. The results suggest that fat decreases hepatic macromolecular adduct formation by inhibiting activation of AFB1 to the epoxide or by enhancing the activity of detoxification pathways.

An immunoblotting diagnostic assay for heartwater based on the immunodominant 32-kilodalton protein of Cowdria ruminantium detects false positives in field sera.

Heartwater, a major constraint to improved livestock production in Zimbabwe, threatens to invade areas which have been previously unaffected. To monitor its spread in Zimbabwe, an immunoblotting diagnostic assay based on the responses of animals to the immunodominant, conserved 32-kDa protein of Cowdria ruminantium was evaluated. In this assay, no false reactions were detected with sera known to be positive and negative, but sera from some cattle, sheep, and goats from heartwater-free areas of Zimbabwe reacted strongly with the 32-kDa protein, suggesting that either these animals had previous exposure to heartwater or they were false positives. To investigate the possibility of previous exposure to heartwater, 11 immunoblot-positive and 6 immunoblot-negative sheep from heartwater-free areas of Zimbabwe were compared regarding their susceptibilities to challenge with C. ruminantium. Prior to challenge, C. ruminantium could not be detected in any sheep by transmission to Amblyomma hebraeum ticks or by the polymerase chain reaction (PCR) conducted with plasma samples. All sheep were equally susceptible to the challenge, and infection was confirmed by brain biopsy, necropsy, PCR, and transmission of C. ruminantium to ticks. Our data suggest that the immunoblot-positive reactions of sera from heartwater-free areas were due not to previous C. ruminantium infection but rather to antigenic cross-reactivity between C. ruminantium and another agent(s) such as Ehrlichia species. In conclusion, the immunodominant 32-kDa protein is not antigenically specific to C. ruminantium and its use in serological diagnosis of heartwater requires reevaluation.

Phosphatidylcholine metabolism in rat liver after partial hepatectomy. Evidence for increased activity and amount of CTP:phosphocholine cytidylyltransferase.

The effect of partial (70%) hepatectomy on phosphatidylcholine (PC) synthesis in rat liver was investigated during the first 4 post-operative days. Between 4 and 96 h after partial hepatectomy, the mass of PC increased from 30% to 80% of sham-operation values, being comparable with the restoration of total liver mass after partial hepatectomy. Relative to control (sham-operation), the incorporation in vivo of [3H]choline into PC was stimulated 2.6-fold at 22 h after partial hepatectomy. Moreover, CTP:phosphocholine cytidylyltransferase (EC 2.7.7.15) activity was significantly enhanced, and the pool size of phosphocholine decreased at 22 and 48 h after partial hepatectomy, whereas the activity of choline kinase (EC 2.7.1.32) was augmented at a later stage of liver regeneration (48 and 96 h). Stimulation of CTP:phosphocholine cytidylyltransferase activity by partial hepatectomy occurred in both the microsomal and cytosolic fractions. The stimulatory effect in the cytosolic fraction was mainly due to an increase in the number of enzyme molecules, as demonstrated by immunotitration of the amount of cytosolic cytidylyltransferase protein.

Biosynthesis and secretion of triacylglycerol in rat liver after partial hepatectomy.

Partially hepatectomized rats were used to investigate the mechanism of fatty-liver development in the regenerating rat liver. After partial hepatectomy the amount of hepatic triacylglycerol increased by almost 4-fold compared with sham-operated rats. The activities of both cytosolic and microsomal phosphatidate phosphohydrolase were enhanced at 12 h after surgery. The activity of diacylglycerol acyltransferase was increased at a later stage of regeneration. Analysis of plasma lipoproteins showed a significant decrease of lipids associated with very-low-density lipoproteins (VLDL). Relative to control, the rate of hepatic triacylglycerol synthesis from [3H]glycerol in vivo was stimulated at 22 h after partial liver resection. However, secretion of glycerol-labelled triacylglycerol in VLDL was the same in control and hepatectomized rats. In cultures of hepatocytes from hepatectomized donor rats, the concentration of triacylglycerol and the biosynthesis of this lipid from [3H]glycerol or from [3H]oleate were enhanced. The secretion of total triacylglycerol into the medium was not affected, resulting in a net accumulation of intracellular triacylglycerol. The rate of secretion of leucine-labelled apolipoproteins B and E associated with VLDL was similar in cell cultures from hepatectomized and sham-operated rats. The results of this study show that the enhancement of the biosynthesis of triacylglycerol in hepatectomized livers is not accompanied by an increase of the secretion of VLDL.

The effect of schistosomiasis on the covalent binding of 2-acetylaminofluorene to mouse liver macromolecules in vivo and in vitro.

The covalent binding of [14C]acetylaminofluorene (AAF) to macromolecules in vivo and in vitro was measured in Schistosoma mansoni-infected and in non-infected mice. Liver microsomes from infected mice demonstrated a 42% decreased capacity to mediate covalent binding of AAF to DNA. In addition, the extent of binding of AAF to liver macromolecules in vivo was generally less in infected than non-infected mice.

Human exposure to aflatoxins in Zimbabwe.

In this communication, we report the detection of aflatoxins in human urine and breast milk. The 2553 urine samples were collected from donors of different ages and sexes at centres throughout Zimbabwe, while 54 breast milk samples were collected from breast feeding mothers. The most predominant aflatoxins found were AFM and AFG. The national average of urine samples contaminated was 6.0 percent. There were, however, some areas in which the extent of contamination was 34 percent. Of the 54 breast milk samples collected, 11 percent were contaminated mainly with AFM.

A survey of urinary aflatoxin in Zimbabwe.

In this study, 1228 urine samples were collected from different centres in Zimbabwe and were analysed for aflatoxin contamination. The urine samples were extracted with chloroform and analysed by thin layer chromatography and high-performance liquid chromatography. The most commonly observed contaminant was aflatoxin M1, at an average concentration of 4.2 ng/ml of urine. Although the national average of urine samples contaminated was 4.3%, there were areas in which up to 10% of the urine samples were contaminated.

The effect of schistosomiasis on the activation of aflatoxin B1.

This study examined activation of aflatoxin B1 (AFB1) in livers of Schistosoma mansoni-infected and noninfected mice by measuring covalent binding of [3H]AFB1 to cellular macromolecules in vivo and in vitro. During a one week time period after AFB1 treatment of animals, maximal binding of [3H]AFB1 to DNA, RNA and protein in liver occurred during the 1-6 hour period after treatment, with less binding throughout of AFB1 to macromolecules of infected mice. Experiments performed in vitro to determine the capacity of liver microsomes to mediate the binding of AFB1 to calf thymus DNA showed that microsomes from infected mice mediated the binding of less [3H]AFB1 to DNA than those from noninfected animals.

Evaluation of 1-(3,4-dichlorophenyl)-4-dimethylaminomethyl-1-nonen-3-one hydrochloride effect on nucleic acid and protein syntheses using murine leukemia L-1210 cells.

Several Mannich bases derived from conjugated styryl ketones were shown to have potent cytotoxicity toward murine leukemia L-1210 cells and Walker 256 carcinosarcoma cells in culture. The most cytotoxic derivative, (E)-1-(3,4-dichlorophenyl)-4-dimethylaminomethyl-1-nonen-3-one hydrochloride, profoundly inhibited the incorporation of tritiated leucine into protein(s) and tritiated deoxythymidine into DNA at concentrations of 0.79-1.32 muM in L-1210 cells. At higher concentrations, incorporation of triated uridine into RNA and tritiated deoxyuridine into DNA was inhibited to a lesser degree. This compound failed to inhibit the enzymes thymidylate synthetase or dihydrofolate reductase up to a concentration of 10-4 M and was ineffective in retarding the growth of the Walker 256 carcinosarcoma in rats.

Syntheses and bioactivities of 1-(hydroxyphenyl)-1-nonen-3-ones and related ethers and esters.

A number of nuclear hydroxy styryl ketones and related compounds were prepared and evaluated for antineoplastic and antimicrobial activities as well as for analgesic, anti-inflammatory, and antianaphylactic properties. The second-order rate constants for the reaction of several esters with hydroxide ion in aqueous dioxane (50% v/v) at 36.9 degrees were determined. The screening results showed that activity against P-388 lymphocytic leukemia was found solely with the ethers and that antimicrobial properties were obtained virtually exclusively with the phenolic derivatives. All compounds showed analgesic properties, except for four that were algesic. While little anti-inflammatory activity was found, several compounds showed some antianaphylaxis.

Synthesis and evaluation of 1-(hydroxyphenyl)-1-nonen-3-ones and related compounds for antineoplastic and antimicrobial activities.

Some 1-(hydroxyphenyl)-1-nonen-3-ones, the corresponding Mannich bases, and O-benzoyl esters were synthesized. Evaluation of these derivatives against murine P-388 lymphocytic leukemia indicated that, while the hydroxyphenyl styryl ketones and related esters were devoid of significant anticancer activities, etherification of the nuclear hydroxyl group gave compounds with a discernible increase in mean survival time. The hydroxyphenyl styryl ketones showed marked potencies against two pathogenic fungi and one, yeast, while the corresponding ethers had diminished activities and the related esters were virtually devoid of antimicrobial activities. Two Mannich bases showed similar spectra of antimicrobial activities as the phenols and, in particular, were active against Trichophyton mentagrophytes and Saccharomyces uvarum.