Prevalence of cdtABC genes encoding cytolethal distending toxin among Haemophilus ducreyi and Actinobacillus actinomycetemcomitans strains.

The aim of this study was to investigate the presence of the three cdtABC genes responsible for production of cytolethal distending toxin (CDT) in Haemophilus ducreyi and Actinobacillus actinomycetemcomitans strains. Of 100 H. ducreyi strains from the culture collection of the University of Göteborg (CCUG), 27 strains with low or intermediate cytotoxic titre (< 1 in 10(4)) and 23 of the remaining isolates with a high cytotoxic titre (> or = 1 in 10(4)) were selected. Twenty-nine strains of H. ducreyi were isolated recently from patients with chancroid and 50 A. actinomycetemcomitans strains from patients with periodontitis. The cytotoxic activity on HEp-2 cells and the presence of cdtABC genes were studied by cytotoxicity assay of bacterial sonicates and PCR with primers specific for individual cdtA, B, and C genes of H. ducreyi in bacterial DNA preparations, respectively. All strains that manifested a cytotoxic titre in sonicate > or = 1 in 100 possessed all the three cdt genes. Eighteen of the 50 strains selected from the culture collection were negative and 32 positive for cdt genes. As all strains with a high cytotoxic titre gave positive PCR results, it can be assumed that the remaining 50 strains, which have high cytotoxic titre, would have been positive as well. Thus, it can be estimated that 82% of the culture collection strains had cdtABC genes. Similarly, 24 (83%) of 29 recent H. ducreyi isolates expressed the CDT activity and displayed all cdtABC genes. Forty-three (86%) of 50 strains of the closely related A. actinomycetemcomitans, expressing a cytotoxic activity > or = 1 in 100, also possessed all three genes. Furthermore, the nucleotide sequence of the cdtABC genes was highly conserved among H. ducreyi strains from different geographic areas. These results indicate that the majority of pathogenic H. ducreyi and A. actinomycetemcomitans strains express a CDT activity encoded by all three cdtABC.

The role of different protein components from the Haemophilus ducreyi cytolethal distending toxin in the generation of cell toxicity.

Cytolethal distending toxin of Haemophilus ducreyi (HdCDT) is a multicomponent toxin, encoded by an operon consisting of three genes, cdtABC. To investigate the role of the individual products in generation of toxicity, recombinant plasmids were constructed allowing expression of each of the genes individually or in different combinations in Escherichia coli and Vibrio cholerae. Expression of all three genes (cdtABC) was necessary to generate toxicity on cells, and no activity was obtained using combinations in which only one or two of the genes were expressed. Of the individual gene products, the CdtA was shown to exist in two forms with an MW of 23 and 17 kDa, respectively. The CdtB protein alone resulted in DNase activity. CdtC purified from both toxic and non-toxic extracts (from strains expressing cdtCAB and cdtC, respectively) had a molecular weight of about 20 kDa and reacted with a CdtC-specific monoclonal antibody. However, the protein isoelectric point (pI) of CdtC from toxic preparations was about 1.5 pH units more basic than from non-toxic ones. Both forms were immunogenic giving rise to toxin-neutralizing antibodies. Toxicity was reconstructed by combining non-toxic cell sonicates from E. coli, expressing CdtA, CdtB and CdtC proteins individually. Only combinations including all three products gave toxicity, indicating that all are actively involved in the generation of toxic activity on cells. The reconstruction resulted in a 1.5 pH unit shift in the PI of CdtC, making it identical to that of the protein isolated from bacteria expressing cdtABC. The results showed that the CdtB component produces DNase activity, but cell toxicity depends on the involvement of the other two components of CDT and is associated with absorption of all three proteins by HEp-2 cells.

Localized pleural plaques and lung cancer.

In a mass chest radiography survey conducted in 1971 for 7,986 residents of three Finnish communities, 604 subjects (7.6%) with pleural plaques but not other asbestos-related radiographic signs were identified. The same number of referents, each individually matched to each plaque carrier on sex, birth year, and community, was selected from among persons in the same source population with no pleural plaques. The two groups were followed for investigation of incidence of lung cancer during 1972-1989. Twenty-eight of those with plaques and 25 referents contracted lung cancer (crude conditional RR = 1.1; CL95 = 0.7, 1.9). The application of the proportional hazards model, with adjustment for sex, age, and residence, resulted in a hazard ratio of 1.1 (CL = 0.6, 1.8). The risk ratio estimate may be biased; hence, the result is inconclusive in regard to the predictive assessment of lung cancer risk among carriers of pleural plaques.

Mortality among sulfide ore miners.

Lung cancer mortality was studied during 1965-1985 in Outokumpu township in North Karelia, where an old copper mine was located. Age-specific lung cancer death rates (1968-1985) were higher among the male population of Outokumpu than among the North Karelian male population of the same age excluding the Outokumpu district (p less than .01). Of all 106 persons who died from lung cancer during 1965-1985 in Outokumpu township, 47 were miners of the old mine, 39 of whom had worked there for at least three years and been heavily exposed to radon daughters and silica dust. The study cohort consisted of 597 miners first employed between 1954 and 1973 by a new copper mine and a zinc mine, and employed there for at least 3 years. The period of follow-up was 1954-1986. The number of person-years was 14,782. The total number of deaths was 102; the expected number was 72.8 based on the general male population and 97.8 based on the mortality of the male population of North Karelia. The excess mortality among miners was due mainly to ischemic heart disease (IHD); 44 were observed, the expected number was 22.1, based on the general male population, and the North Karelian expected number was 31.2 (p less than .05). Of the 44 miners who died from IHD, 20 were drillers or chargers exposed to nitroglycerin in dynamite charges, but also to several simultaneous stress factors including PAHs, noise, vibration, heavy work, accident risk, and working alone. Altogether 16 tumors were observed in the cohort. Ten of these were lung cancers, the expected number being 4.3. Miners who had died from lung cancer were 35-64 years old, and had entered mining work between 1954 and 1960. Five of the ten lung cancer cases came from the zinc mine (1.7 expected). Three of them were conductors of diesel-powered ore trains. The slight excess mortality from lung cancer could be explained by exposure to radon daughters and by the combined effect of silica dust and diesel exhaust gases in the zinc mine.

Asbestos disease in Finland.

Finland was the world's main producer of anthophyllite asbestos between 1918 and 1975. The total amount of mined anthophyllite asbestos was 350,000 tons of which 230,000 tons were exported. Anthophyllite quarry workers who had worked at least five years between 1936 and 1972 showed the highest risk of asbestosis, in comparison with other groups of asbestos workers.

A health survey of granite workers in Finland: radiographic findings, respiratory function, hearing, electric sensory thresholds of the fingers and subjective symptoms.

Results from a health survey of 777 granite workers, exposed to quartz dust, noise and/or machine vibration, and 122 unexposed "controls" are reported. The survey included chest X-rays, pulmonary function tests, a hearing examination, the determination of the electric sensory thresholds of the fingers, and questionnaires on the subjective symptoms of the respiratory system and upper limbs. The exposed workers' radiographic findings (except for a 2.6 % rate of silicosis), respiratory functions, and sensory thresholds did not differ significantly from those of the controls. Granite drillers had a high prevalence of hearing defects. Respiratory symptoms were common among drillers and sandblasters, and subjective symptoms of the upper limbs due to vibration occurred among the granite dressing workers who used pneumatic hammers. Both primary and secondary measures of prevention are strongly recommended for the granite industry, primarlily for quartz dust exposure and its health effects.