S. gallolyticus Aortic Valve Endocarditis with Mitral Valve Leaflet Aneurysm.

S. gallolyticus is one of the pathogenic agents of endocarditis, and mitral valve aneurysm is a rare but potentially devastating complication. We present a case of S. gallolyticus aortic valve endocarditis with concomitant anterior mitral valve leaflet aneurysm. Patient underwent surgery before aneurysm perforation, and postoperative course was uneventful. Time of surgery is crucial to avoid severe complications due to aneurysm rupture.

Cytogenetic and immunofluorescence analysis of benzo[a]pyrene-DNA adduct formation and chromosome damage in larval brain neuroblasts of Drosophila melanogaster.

Recently we have evaluated the relationship between benzo[a]-pyrene(BaP)-DNA adducts, determined by 32P-postlabelling, and clone frequencies in the somatic mutation and recombination test (SMART) in Drosophila melanogaster. Following that study we proceeded to characterise further the mechanism of induction of genetic damage in vivo by BaP in Drosophila by cytogenetic analysis of larval brain neuroblasts. Third stage larvae were treated with 4 and 10 mM BaP for 24, 48 or 72 h. In all cases, the larvae were killed 72 h after the beginning of treatment, entailing 48, 24 or 0 h post-treatment recovery in BaP-free medium, respectively. At the end of the treatment the following data were collected: (i) the types and levels of chromosome aberrations in neuroblast metaphase and anaphase nuclei; (ii) the distribution and level of BaP-DNA adducts, revealed by indirect immunofluorescence in neuroblast nuclei using an anti-(BaP-DNA) antibody. The results indicate that BaP induces chromosome breaks, deletions and exchanges in this system. In particular, chromosome exchanges decrease as the post-treatment recovery time increases, and the dynamics of breaks and deletions appear to be inversely related to those of the exchanges. This suggests that exchanges may require few preconditions to occur and are thus expressed soon after treatment. Chromosome breaks and deletions could require multiple single events before the actual damage is expressed (even some cell divisions away from the end of treatment). The immunofluorescence analysis suggests that BaP-DNA adducts are more abundant in the heterochromatin of the neuroblast nuclei.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between benzo(a)pyrene-DNA adducts and somatic mutation and recombination in Drosophila melanogaster.

The evaluation of the relationship between the dose to DNA of a mutagen/carcinogen and in vivo somatic cell mutagenesis may provide information on the mechanisms leading to induced mutational events. This can be achieved, for example, by coupling test systems that permit the detection of somatic mutation and recombination on the basis of phenotypic changes in cuticular structures of Drosophila melanogaster, with methods for the quantitation of carcinogen-DNA adducts such as the 32P-postlabeling technique. In this article, we evaluate the quantitative relationship between BaP-DNA adduct formation, determined by 32P-postlabeling, and the induction of mutant cells in the wing marker version of the somatic mutation and recombination test (SMART) in Drosophila melanogaster. The total single clones in the trans-heterozygous mwh/flr3 flies show a linear relationship with the BaP-DNA adduct levels, suggesting a single hit mechanism for the genetic damage giving rise to this type of clones. In contrast, the twin clones (which are of recombinational origin) display a linear-quadratic relationship with the adduct levels, suggesting that multiple hits may be involved in generating these clones. The total single clones in the mwh/TM3, Ser flies (in which mitotic recombination is suppressed) show a logarithmic relationship with the adduct levels. The discussion of these data in terms of the pathways that may be involved in the repair of the BaP-DNA adducts leads to the suggestion that in Drosophila melanogaster the repair of Bap metabolite-DNA adducts in somatic cells may proceed, in large part, via post-replicative recombinational repair.

[The urinary mutagenicity test in monitoring exposure to aromatic polycyclic hydrocarbons in workers in the aluminum industry]. / Test di mutangenicità urinaria nel monitoraggio dell'esposizione ad idrocarburi policiclici aromatici di lavoratori dell'industria di alluminio.

The sensitivity of 3 urinary mutagenicity tests was assayed: the plate test, the fluctuation test and the micropreincubation test, in order to assess their possible use in monitoring human exposure to polycyclic aromatic hydrocarbons (PAH). Urine samples from workers of an anode production plant exposed to coal tar and from psoriatic patients undergoing treatment with coal-tar ointments were tested for mutagenic activity on strain TA98 Salmonella typhimurium, in the presence of the microsome fraction and deconjugating enzymes. Parallelly, the urinary concentration of PAH metabolites or one of their trace metabolites, 1-hydroxypyrene, was determined. Increased levels of PAH metabolites were observed in the urine of anode production workers after a work shift compared with controls. Results of the plate test and the fluctuation test performed on urine of exposed subjects, both smokers and nonsmokers, showed mutagenicity values similar to the controls. Much higher 1-hydroxypyrene concentrations were found in the urine of psoriatic patients treated with coal tar than in post-shift urine of anode production workers. The urine of the former was also mutagenic in the 3 mutagenicity tests used. The minimum mean dose of PAH metabolites was calculated, expressed as quantity of 1-hydroxypyrene, that would give a mutagenic response in the 3 tests: the micropreincubation test was found to be about 100 times more sensitive than the plate test and about 30 times more sensitive than the fluctuation test. The theoretical minimum urinary concentration of 1-hydroxypyrene detectable by each test was determined: the micropreincubation test was 15 times more sensitive than the plate test and 7 times more sensitive than the fluctuation test.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of benzo[a]pyrene-diol-epoxide-DNA adducts in white blood cells of psoriatic patients treated with coal tar.

An enzyme-linked immunosorbent assay (ELISA) was used to detect BPDE-DNA adducts in white blood cells of 23 psoriatic patients undergoing clinical coal tar therapy. Ten of these patients were reanalyzed 2-5 months after the end of the coal tar treatments. The results show that the mean adduct level during the treatment period was 0.26 +/- 0.16 fmole BPDE/micrograms DNA (7.7 +/- 4.9 adducts/10(8) nucleotides), while 2-5 months later the mean adduct level had decreased significantly (P less than 0.005) to 0.11 +/- 0.08 fmole BPDE/micrograms DNA (3.3 +/- 2.4 adducts/10(8) nucleotides). No relationship could be ascertained between the level of exposure and the amount of BPDE-DNA adducts. In addition, no difference in the level of DNA adducts was found between smoking and non-smoking patients.

Direct involvement of human gastric mucosa in the activation of alimentary pro-carcinogens.

Exposure to N-nitroso-compounds and aromatic amines, xenobiotics which require an activation in order to exert their genotoxic potential, is causally associated with gastric cancer. We evaluated the capacity of microsome-containing fractions from human gastric mucosa to activate two model carcinogenic compounds. A 9,000 x g supernatant (S9) was obtained from gastric mucosal specimens and, for comparison, from human liver and aroclor-induced and non induced rat liver. The capacity of the S9 to activate N-nitrosopyrrolidine (NPY) and 2-aminofluorene (2AF) to mutagenic metabolites was tested in the Ames/Salmonella reversion assay, while dimethylnitrosamine (DMN) and aminopyrine (AP) demethylation activities were spectrophotometrically evaluated by using an enzymatic assay of the amount of formaldehyde released following the enzymatic demethylation of the corresponding substrates. Results indicate that human gastric S9 fractions may activate 2AF to a genotoxic derivative and are characterized by DMN and AP demethylase activities higher (p < 0.05) than those of human liver, when expressed in mg/protein (p < 0.05). Although the parameters evaluated can only be considered as a partial measure of the general activating capacity toward dietary and environmental procarcinogens, these results suggest that human gastric mucosa may be directly involved in the metabolic activation of these compounds to mutagenic/carcinogenic species.

Biological monitoring of human exposure to coal tar. Urinary mutagenicity assays and analytical determination of polycyclic aromatic hydrocarbon metabolites in urine.

The mutagenicity of urine extracts from anode plant workers exposed to coal tar pitch volatiles and non-smoking psoriatic patients treated with coal tar applications and UV light (Goeckermann regimen), was determined by the plate incorporation assay and the fluctuation test employing Salmonella typhimurium strain TA98 in the presence of rat liver post-mitochondrial fractions and deconjugating enzymes. The levels of total polycyclic aromatic hydrocarbons (PAHs) and of a marker metabolite of pyrene (1-hydroxypyrene) were determined in the urine of the same subjects. Both the occupational and in particular the therapeutic exposure to coal tar resulted in clear increases in urinary levels of PAH metabolites as compared to unexposed subjects. The level of 1-hydroxypyrene in the urine samples was comparable to or even greater than the corresponding level of total PAHs, indicating a poor recovery of PAH metabolites for this method. Following treatment with coal tar, most of the psoriatic patients excreted clearly increased levels of mutagens in their urine, while non-smoking anode plant workers showed no increase in urinary mutagenicity. The minimum levels of PAH metabolites corresponding to a significant increase in urinary mutagenicity varied from sample to sample, presumably depending on interfering factors present in different amounts in the extracts. Nonetheless the urine samples which were clearly mutagenic presented elevated levels of PAH metabolites, suggesting that the mutagenicity assays lack sufficient sensitivity to allow their application in the biological monitoring of most occupational exposures to coal tar.

Chromosomal alterations in peripheral blood lymphocytes, urinary mutagenicity and excretion of polycyclic aromatic hydrocarbons in six psoriatic patients undergoing coal tar therapy.

Six male non-smoking subjects treated for psoriasis with topical applications of pure coal tar or 4% coal tar-containing ointment were examined in order to assess the genotoxic risk associated with this type of therapy. Mutagenicity in urine samples collected before and during the coal tar therapy was evaluated in the plate incorporation assay on Salmonella typhimurium strain TA98 in the presence of S9 mix and beta-glucuronidase. Total urinary polycyclic aromatic hydrocarbon (PAH) levels were evaluated in parallel by high resolution gas chromatography/mass spectrometry. In addition, sister chromatid exchanges and chromosomal aberrations were also analysed in peripheral blood lymphocytes collected before, during and after the end of the coal tar applications. The results suggest that urinary mutagenicity levels as well as the frequencies of chromosome aberrations and sister chromatid exchanges in lymphocytes are related to the levels of exposure to coal tar. Moreover the kinetics of repair of chromosome damage in relation to different exposure levels and the capacity of the urinary mutagenicity assay to correctly identify the exposure to significant levels of PAH are discussed.

Induction of SOS response in Escherichia coli strain PQ37 by 16 chemical compounds and human urine extracts.

The SOS Chromotest on Escherichia coli strain PQ37 was used to detect DNA damage induced by 16 chemical compounds and urine samples from smokers and a non-smoking psoriatic patient treated with mineral coal tar. The results confirmed the strong SOS inducing activity of 2-aminoanthracene and benzo[a]pyrene with metabolic activation and N-methyl-N'-nitro-N-nitrosoguanidine, mitomycin C and 4-nitroquinoline-N-oxide without metabolic activation. A weaker response in the absence of microsomal enzymes was observed with hydroxyurea (only at high doses) and the soluble Cr(VI) compounds potassium chromate and potassium dichromate. No effect was observed with ampicillin, cadmium chloride, cyclophosphamide, griseofulvin, the insoluble Cr(VI) compound lead chromate, the soluble Cr(III) compounds chromium nitrate, chromium chloride, chromium potassium sulphate, and the chelating agent sodium nitrilotriacetate. Among the Cr(III) compounds only chromium acetate produced a low but significant increase of SOS inducing activity. Solubilization by nitrilotriacetate of genotoxic Cr(VI) from insoluble lead chromate was observed, whereas no interaction occurred between nitrilotriacetate and the soluble Cr(VI) and Cr(III) compounds. Using urinary XAD-2 extracts, we found the SOS Chromotest poorly sensitive to the mutagens present in urine from tobacco smokers which, on the other hand, were detected by the gene mutation assay in Salmonella typhimurium (Ames test). A urine sample obtained from a psoriatic patient, therapeutically treated with mineral coal tar, had a significant SOS inducing activity with and even without metabolic activation, whereas in the Ames test it was active only in the presence of metabolic activation.(ABSTRACT TRUNCATED AT 250 WORDS)

Biological monitoring of human exposure to coal tar. Urinary excretion of total polycyclic aromatic hydrocarbons, 1-hydroxypyrene and mutagens in psoriatic patients.

Three methods for the biological monitoring of human exposure to coal tar were compared. Levels of 1-hydroxypyrene(1-OH PYR), polycyclic aromatic hydrocarbons (PAH) and mutagens (Ames plate incorporation assay using Salmonella typhimurium strain TA98 in the presence of S9 and beta-glucuronidase) were determined in urinary samples from psoriatic patients undergoing topical treatment with mineral coal tar. A single sample of urine with a high content of PAH was diluted with urine of nonexposed, non-smoking subjects in order to obtain nine samples with a decreasing content of PAh metabolites. Mutagenicity of the extracts was detectable down to the dilution corresponding to a content in 1-OH PYR of about 50 micrograms/g creatinine and total PAH of 7 micrograms/g creatinine. In a second phase the three indicators of exposure to PAH were compared in 16 urinary samples from four psoriatic patients. The total PAH levels determined by the acidic deconjugation/reduction method were confirmed to be nearly always lower than the corresponding levels of 1-OH PYR alone. Most of the extracts were mutagenic, however, some of the samples with a high content in PAh metabolites were not mutagenic. In all the urinary samples analyzed the excretion of 1-OH PYR was markedly greater than in control subjects. 1-OH PYR and urinary mutagenicity levels were well correlated. The present data suggest that both the determination of mutagenicity and 1-OH PYR in urine may be used to monitor occupational exposure to PAH, the latter method being cheaper and of greater specificity and sensitivity.

Mutagenic activity and polycyclic aromatic hydrocarbon levels in urine of humans exposed to therapeutical coal tar.

The urinary mutagenicity and the excretion of polycyclic aromatic hydrocarbons (PAH) in three non-smoking male patients, treated for psoriasis with cutaneous applications of crude coal tar, were analysed. Mutagenicity of the urinary extracts was measured by the plate incorporation assay using Salmonella typhimurium strains TA 98 and TA 100 in the presence of liver S9 fraction from Aroclor-induced rats with or without beta-glucuronidase. After concentration, hydrolysis and reduction of the urine sample, PAH levels were measured by high resolution gas chromatography/mass spectrometry. Following cutaneous treatment with coal tar, the urine of all three subjects showed noticeable levels of PAH and/or metabolites and marked mutagenicity both on strain TA 98 and TA 100 in the presence of S9 fraction. The addition of beta-glucuronidase increased the mutagenicity of the urinary extracts, the maximum values being attained on strain TA 100 in the presence of both microsomal fraction and deconjugating enzymes. The mutagenicity of urinary extracts from subjects treated therapeutically with crude coal tar was correlated (r = 0.788, P less than 0.01) with the total PAH levels in their urine. The PAH excreted in urine were mainly low molecular weight compounds, while benzo[a]anthracene was present in scarce amounts and the excretion of benzo[a]pyrene did not increase following the cutaneous exposure to the crude coal tar.

Mutagenic activity and polycyclic aromatic hydrocarbon levels in urine of workers exposed to coal tar pitch volatiles in an anode plant.

The mutagenicity of urinary extracts and the excretion of PAH from workers occupationally exposed to coal tar pitch volatiles in an anode plant were analyzed. Mutagenicity of the urinary extracts was measured by means of the plate test using S. typhimurium strain TA 98. After concentration, hydrolysis and reduction of the urine samples, PAH levels were measured by high resolution gas chromatography/mass spectrometry. No significant difference was found in the mutagenicity of the urinary extracts of non-smokers occupationally exposed to PAH as compared with the controls. Low PAH concentrations were found in the urine of the exposed subjects, which lends further support to the negative results obtained with the Ames' test. The increase of urinary PAH excretion, in relation to occupational exposure, was mainly due to the less mutagenic, low molecular weight compounds.