Viral load and humoral immune response in association with disease severity in Puumala hantavirus-infected patients--implications for treatment.

Hantaviruses are the causative agents of haemorrhagic fever with renal syndrome (HFRS) in Eurasia and of hantavirus cardiopulmonary syndrome (HCPS) in the Americas. The case fatality rate varies between different hantaviruses and can be up to 40%. At present, there is no specific treatment available. The hantavirus pathogenesis is not well understood, but most likely, both virus-mediated and host-mediated mechanisms are involved. The aim of the present study was to investigate the association among Puumala hantavirus (PUUV) viral RNA load, humoral immune response and disease severity in patients with HFRS. We performed a study of 105 PUUV-infected patients that were followed during the acute phase of disease and for up to 1-3 months later. Fifteen of the 105 patients (14%) were classified as having moderate/severe disease. A low PUUV-specific IgG response (p <0.05) and also a higher white blood cell count (p <0.001) were significantly associated with more severe disease. The PUUV RNA was detected in a majority of patient plasma samples up to 9 days after disease onset; however, PUUV RNA load or longevity of viraemia were not significantly associated with disease severity. We conclude that a low specific IgG response was associated with disease severity in patients with HFRS, whereas PUUV RNA load did not seem to affect the severity of HFRS. Our results raise the possibility of passive immunotherapy as a useful treatment for hantavirus-infected patients.

Vitamin A (retinol) status in the Gunn rat. The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Vitamin A (retinol) status and the effect of a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 10 micrograms X kg-1, on vitamin A in the liver and serum, and on the hepatic UDP-glucuronosyltransferase (UDPGT) (EC 2.4.1.17) activity, were studied in heterozygous (GW) and homozygous (GG) Gunn rats. 1) Data from vitamin A analyses demonstrate that the amount of vitamin A stored in the liver of untreated Gunn rats is of the same magnitude as that of Sprague-Dawley rats. 2) The retinol content in the liver of both GG and GW rats was reduced to about 50% by TCDD-treatment. 3) Retinol levels in serum were found to be variable and no significant effect due to TCDD could be observed. 4) No correlation between the TCDD-induced reduction of vitamin A and the induction of UDPGT activity by TCDD could be demonstrated in this study. The vitamin A reduction caused by TCDD was considerably less in the Gunn rat than in the Sprague-Dawley rat, and the results indicate that the Gunn rat is more resistant to TCDD than other strains of rat. TCDD-induced reduction of liver vitamin A seems to some extent to correlate with TCDD-toxicity in different strains of rat. The specific properties of the Gunn rat and its relatively high resistance to TCDD make it a valuable tool in studies about the mechanism of TCDD-toxicity.

Chlorinated phenols: occurrence, toxicity, metabolism, and environmental impact.

Pentachlorophenol and the lower chlorinated phenols, tetra- and trichlorophenols, have gained an increasing use as fungicides, herbicides, insecticides, and precursors in the synthesis of other pesticides since the early 1930s. World-wide production totals about 200,000 tons. Production and use of chlorinated phenols have caused industrial hygiene problems but, otherwise, have not been recognized to create more than limited environmental problems. The introduction of modern analytical techniques, however, has revealed the ubiquitous occurrence of chlorophenols in the environment, and the discovery of chlorinated dimers, such as dibenzo-p-dioxins and dibenzofurans, as impurities in commercial chlorophenol formulations, has made a reevaluation of the chlorinated phenols necessary. The present article reviews recent studies on the toxicity and metabolism in mammals and aquatic organisms and the degradation of the chlorophenols under various conditions in the environment. Finally, the hazards of burning of chlorophenol wastes are discussed, as well as health considerations with regard to humans and the environment.

Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the in vivo and in vitro dechlorination of pentachlorophenol.

The metabolism of pentachlorophenol has been studied in the rat after pretreatments with phenobarbital, 3-methyl cholanthrene or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In addition to the previously identified metabolite, tetrachloro-p-hydroquinone, trichloro-p-hydroquinone has been identified in urine as a metabolite. The formation of the latter represents a type dechlorination different from that of the formation of tetrachlorohydroquinone. The inducing agents, 3-methylcholanthrene and TCDD have similar effects on the dechlorination and increase the formation of tetrachloro-p-hydroquinone more pronounced than does phenobarbital. In contrast to phenobarbital they also increase the formation of trichloro-p-hydroquinone and the total elimination of pentachlorophenol and its metabolites. The in vivo findings are supported by in vitro studies with microsomes from rats pretreated with phenobarbital or TCDD. Use of the inhibitor beta-diethylaminoethyl-diphenyl propylacetate (SKF 525-A) in vitro showed a more pronounced inhibition on microsomes from phenobarbital-treated rats than on microsomes from untreated or TCDD-treated rats. Gas chromatography-mass spectrometry have been used for the identification and quantification of pentachlorophenol and its metabolites.

Metabolism of pentachlorophenol in vivo and in vitro.

Pentachlorophenol has earlier been shown to be metabolized in mammals to tetrachloro-p-hydroquinone. The metabolite possesses pronounced inhibitory activity on bacterial beta-glucuronidase but not on beta-glucuronidase from liver. Indirect evidence for the occurrence of both pentachlorophenol and tetrachloro-p-hydroquinone as conjugates with glucuronic acid in the urine from pentachlorophenol-treated rats is now presented. Bovine liver beta-glucuronidase has been utlizied to split the conjugates present. The in vivo metabolism of pentachlorophenol has also been studied in rats treated with phenobarbital and beta-diethylaminoethylidiphenyl propylacetate (SKF 525-A). In vitro metabolism has been studied using liver microsomes from rats pretreated with pehnobarbital. Quantitative analysis of the compounds occurring in extracts of urine or extracts from the microsomal incubates was performed by means of mass fragmentography. Pretreatment with phenobarbital increased the metabolism of pentachlorophenol to tetrachloro-p-hydroquinone both in vivo and in vitro. SKF 525-A, however, inhibited the metabolism in vitro but enhanced the metabolism in vivo when given less frequently than every 6th h. Dechlorination of pentachlorophenol is mediated by microsomal enzymes that can be induced by phenobarbital. SKF 525-A does not inhibit the dechlorination in vivo but does so in vitro.

Vitamin A (retinol) status in the rat after a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The effect of a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 10 microgram . kg-1, on the status of vitamin A (retinol) was studied in rats during an eight week period after administration. Retinol was determined spectrophotometrically after separation by means of high pressure liquid chromatography. In the liver of control animals the total storage and the concentration of retinol were found to increase linearly with time whilst in the TCDD-exposed animals both the concentration and total storage remained essentially unchanged. Differences in the storage levels were evident after 4 days, and after 8 weeks the treated animals had a total liver storage corresponding to about 30% of the controls. Retinol levels in serum were significantly higher in the treated animals after week 1 and 2.

Comparison between the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin and six other compounds on the vitamin A storage, the UDP-glucuronosyltransferase and the aryl hydrocarbon hydroxylase activity in the rat liver.

The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8- tetrabromodibenzo -p-dioxin ( TBrDD ), 5-chloro-2-(2,4-dichlorophenoxy)phenol (3-Cl- predioxin ) 4,5,6-trichloro-2-(2,4-dichlorophenoxy)phenol (5-Cl- predioxin ), toxaphene, 3-methylcholanthrene (3-MC) and phenobarbital (PB) on the vitamin A storage, UDP-glucuronosyltransferase (UDPGT) and aryl hydrocarbon hydroxylase (AHH) activities in the liver of Sprague-Dawley rats was investigated. Vitamin A was determined as retinol by high pressure liquid chromatography. UDPGT was measured with p-nitrophenol as an aglycone and AHH with 3,4-benzopyrene as a substrate. Both in TCDD- and toxaphene-treated animals a reduced body weight gain was recorded, but no other overt signs of toxicity were seen in this study. Both the concentration and the total amount of hepatic retinol was significantly reduced in TCDD-, 3-MC-, PB- and TBrDD -treated animals. These compounds were also those which gave the most significant enzyme induction as regards the UDPGT and AHH activities. However, the reduction of hepatic retinol caused by these compounds did not correlate with the enzyme activities studied. When compared on a molecular basis, TCDD and TBrDD were in the order of several magnitudes more potent as reducers of hepatic retinol and likewise as enzyme inducers.

Inhibition of beta-glucuronidase by chlorinated hydroquinones and benzoquinones.

An earlier study of the metabolism of pentachlorophenol has shown that a metabolite, tetrachloro-p-hydroquinone, possessed pronounced inhibitory action on the activity of beta-glucuronidase from bacterial origin. Several other chlorinated hydroquinones and benzoquinones have now been studied with regard to their ability to inhibit beta-glucuronidase of various origin in vitro and in vivo. All the studied chlorinated hydroquinones and benzoquinones were found to be potent inhibitors of beta-glucuronidase of bacterial origin. D-glucaric acid-1.4-lactone was included for comparison and was found to be less active than the other studied compounds. The inhibition was found to be competitive in nature. No inhibitory effect of the benzo- and hydroquinones studied in vitro or in vivo could be demonstrated on beta-glucuronidase from livers. The result calls for precaution when using bacterial beta-glucuronidase to split urinary conjugates of glucuronic acid.

Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the hepatic storage of retinol in rats with different dietary supplies of vitamin A (retinol).

The effect of various dietary sources of vitamin A on liver storage of retinol has been investigated in Sprague-Dawley rats treated with single oral doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD): 0,0.1,1.0, or 10 microgram.kg-1. Each dose group consisted of 3 subgroups, each comprising 10 rats which received a diet with normal, low or high retinol content. The animals were killed 4 weeks after TCDD administration. Analyses of retinol were performed by high pressure liquid chromatography and glucuronosyltransferase activities were determined spectrophotometrically. A dose-dependent decrease in hepatic storage of retinol was evident. The high retinol diet did not fully compensate for the reduction caused by the highest TCDD-dose. Glucuronosyltransferase activity increased directly in relation to the TCDD-dose but in inverse proportion to the retinol content of the diet.