Induction of chromosomal aberrations and spindle disturbances in Chinese hamster epithelial liver cells in culture by pyrene and benzo[a]pyrene quinones

Regioisomers of pyrene and benzo[a]pyrene quinones were testedfor their ability to induce structural and numerical aberrationsand spindle disturbance in Chinese hamster epithelial liver(CHEL) cells in culture. All quinones tested were clastogenicPyrene-1,8-quinone (P-1,8-Q) and benzo[a]pyrene–3,6–quinone(BP-3,6-Q) induced strikingly high levels of triradials. Inaddition, dicentrics and ring chromosomes were very common inBP-3,6-Q-treated cultures. Isomers of these compounds, pyrene-1,6-quinone(P-1,6-Q) and benzo[a]pyrene-1,6-quinone (BP-1,6-Q), inducedunobtrusive patterns of chromosomal aberrations. We suspectthat the P-1,8-Q and BP-3,6-Q moieties bound to the DNA werestill reactive, and formed crosslinks and/or underwent redoxcycling leading to high local concentrations of reactive oxygenspecies. In addition, P-1,8-Q and BP-3,6-Q induced c-mitoses,hyperdiploidies and polyploidies, in particular endoreduplications.These effects were not seen with the other two test compounds,or they were only detected at the highest concentrations used,which were strongly cytotoxic (c-mitoses with P-1,6-Q, polyploidieswith BP-1,6-Q). ⁶To whom correspondence should be addressed at: European Centre for the Validation of Alternative Methods (ECVAM), Joint Research Centre (JRC), TP58O, 1–21020 Ispra, Italy     

Comparison of three immunoassays for diagnosing sensitization to latex in children with spina bifida

As natural rubber latex (latex) has become more widespread in our environment, physicians have become increasingly aware of the problem of possible allergic reactions. Many fatal and near-fatal incidents have been reported (mainly during surgery) (1—3) and data has been published on groups frequently exposed to latex, such as patients with spina bifida (4—9), healthcare professionals (10—12) and occupationally exposed persons (13). The incidence of latex allergy in children seems to be increasing (14). Tests are therefore needed which can reliably detect sensitization to latex. Our aim was to compare the diagnostic accuracy of three commercial immunoassays for measuring specific IgE in serum to latex.     

An anti-CD44 antibody does not enhance engraftment of DLA-identical marrow after low-dose total body irradiation

920 cGy total body irradiation (TBI) is adequate for consistently successful engraftment of marrow from dog leukocyte antigen (DLA)-identical littermates; however, the dose is inadequate to ensure a marrow graft from DLA-nonidentical unrelated donors. Such mismatched grafts are successful only after 1800 cGy, given in three fractions. While anti-T-cell reagents enhance engraftment of DLA-identical littermate marrow after 920 cGy, they fail to be effective in the DLA-nonidentical setting. However, a monoclonal antibody (mAb) to CD44, S5, was found to be very effective in enhancing engraftment of DLA-nonidentical marrow. The current study asked whether mAb S5 was also effective in the setting of DLA-identical littermate transplants. To this purpose, the TBI dose was lowered to 450 cGy, a dose after which 70% of such grafts failed. Four dogs were treated with antibody S5, 0.2 mg/kg on days −7 though −2 (per previously published protocol), given 450 cGy TBI followed by marrow grafts from their DLA-identical littermates. All four dogs rejected their grafts; two of these died from marrow aplasia, and two survived with endogenous marrow recovery. This result was not statistically significantly different from that in 17, historical (n = 5) and concurrent (n = 12), control dogs where 11 of 17 animals rejected. Even if ten experimental animals were transplanted and all six remaining dogs engrafted, the results still would not have been significantly different from control. This result is in contrast to the successful engraftment promoted by pretreatment with antibody S5 of DLA-nonidentical unrelated dogs, consistent with the notion that different host cells are involved in graft rejection in the two disparate histocompatibility settings.     

Isolated and combined effects of prolonged exposures to noise and whole-body vibration on hearing,vision and strain

Summary This study was carried out in order: (1) to examine the effects of isolated and combined prolonged exposures to noise and whole-body vibration on hearing, vision and subjectively experienced strain, and (2) to check the combined effects with repeated exposures. Six male subjects were exposed twice to noise (N) at 92 dBA, whole-body vibration (V) in the Z-axis at 4 Hz and 1.0 ms^–2 rms, and noise and vibration (NV) for 90 min with each condition. Temporary threshold shifts of hearing (TTS) and their integrals (ITTS) were measured at 4, 6, 10, and 12 kHz. Visual acuity was examined by means of a very sensitive test. Cross-modality matching (CMM) of the handgrip force was used to judge the subjectively experienced strain. NV induced a clear tendency of higher TTS and ITTS than N, with several significant differences most pronounced at 10 kHz. With repeated exposures, the effect of NV decreased, while the reactions to N and V remained unchanged. The individual reactions to NV differed. The influence of the duration of exposures on vision depended on the condition; N caused time-dependent changes, whereas V did not. CMM-data increased with the duration of the exposure during V and NV. N was generally judged to be more straining than V; NV caused higher strain than V during the first 30 min of exposure only. Correlations between different effects suggest certain links between them. Additionally, less motivation — daily obtained by a questionnaire — often correlated with higher ITTS during N and NV. The results also illustrate the combined effects on the individual susceptibility, repetition of exposure, the kind of response, and, possibly, the actual psychic state.Abbreviations CMM cross-modality matching - MVC maximum voluntary contraction force - N exposure condition: noise level 92dBA, no whole-body vibration - NV exposure condition: combined exposure to noise with a level of 92 dBA and wholebody vibration with 4 Hz, 1 ms^–2 rms - V exposure condition: whole-body vibration with 4 Hz, 1 ms^–2 rms - TTS temporary threshold shift - ITTS integral of temporary threshold shift - WBV whole-body vibration in the common sense This work was done in the Temporary International Research Team on Combined Effects of Noise and Vibration of the Council of Mutual Economic Assistance of the Socialist Countries. The authors gratefully acknowledge the help and assistance of L.-M. Brumm, Y. Bening, M. Godau, G. Weber, and R. Vizcaino.     

Metabolism of the bay-region diol-epoxide of chrysene to a triol-epoxide and the enzyme-catalysed conjugation of these epoxides with glutathione

Metabolic activation of chrysene in mouse skin appears to involver-1,t-2-dihydroxy-t-3,4-oxy-1,2,3,4-tetrahydrochrysene (anti-chrysene-1,2-diol 3, 4-oxide) and 9-hydroxy-r-1,t-2-dihydroxy-t-3, 4-oxy-1,2, 3, 4-tetrahydrochrysene (anti-9-OH-chrysene-1, 2-diol 3,4-oxide). The enzyme-catalysed conjugation of these epoxideswith [³⁵S]glutathione has been studied in experiments in whichthe glutathione conjugates were separated by h.p.l.c. and examinedby fluorescence spectrophotometry. Both anti-chrysene-1, 2-diol3, 4-oxide and anti-9-OH-chrysene-1, 2-diol 3, 4-oxide formedconjugates nonenzymically and both were shown to be substratesfor rat liver glutathione transferases. When anti-chrysene-1,2-diol 3, 4-oxide was incubated with [³⁵S]glutathione and arat liver microsomal metabolizing system, glutathione conjugateswith h.p.l.c. and fluorescence spectral characteristics identicalto those of conjugates formed from both anti-chrysene-1, 2-diol3,4-oxide and anti-9-OH-chrysene-1, 2-diol 3, 4-oxide were detected.This finding provides evidence that anti-chrysene-1, 2-diol3, 4-oxide can be further metabolized to the triolepoxide, anti-9-OH-chrysene-1,2-diol 3, 4-oxide by rat liver microsomal systems.     

Pediatric prehospital care in urban and rural areas

There are limited data concerning pediatric prehospital care, although pediatric prehospital calls constitute 10% of emergency medical services activity. Data from 10,493 prehospital care reports in 11 counties of California (four emergency medical services systems in rural and urban areas) were collected and analyzed. Comparison of urban and rural data found few significant differences in parameters analyzed. Use of the emergency medical services system by pediatric patients increased with age, but 12.5% of all calls were for children younger than 2 years. Calls for medical problems were most common for patients younger than 5 years of age; trauma was a more common complaint in rural areas (64%, P = .0001). Frequency of vital sign assessment differed by region, as did hospital contact (P less than .0001). Complete assessment of young pediatric patients, with a full set of vital signs and neurologic assessment, was rarely performed. Advanced life support providers were often on the scene, but advanced life support treatments and procedures were infrequently used. This study suggests the need for additional data on which to base emergency medical services system design and some directions for education of prehospital care providers.     

Metabolic activation to a mutagen of 3-hydroxy-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene, a secondary metabolite of benzo[a]pyrene

3-Hydroxy-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (3-OH-BP-7,8-diol)wag isolated from arylsulfatase/ß-glucuronidase-treatedbile of rats to which 3-hydroxybenzo[a]pyrene (3-OH-BP) hasbeen administered. This triol was investigated for mutagenicityin Salmonella typhimurium (reversion to histidine prototrophyof strains TA 97, TA 98, TA 100 and TA 1537) and in V79 Chinesehamster cells (acquisition of resistance to 6-thioguanine).When no exogenous metabolizing system was added the triol wasinactive, while 3-OH-BP showed weak mutagenic effects with allfour bacterial strains. In the presence of NADPH-fortified postmitochondrialsupernatant fraction (S9 mix) of liver homogenate from Aroclor1254-treated rats, the mutagenicity of 3-OH-BP was potentiated,and the triol was activated to a mutagen(s). In the presenceof S9 mix, the triol was 5—18 times more mutagenic than3-OH-BP in strains TA 97, TA 100 and TA 1537, but both compoundsshowed similar mutagenic potencies with strain TA 98. Thesestrain differences strongly suggest that the mutagenicity of3-OH-BP in the S9 mix-mediated test was not exclusively dueto metabolites of 3-OH-BP-7, 8-diol. Trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene(BP-7,8-diol), like the triol, showed mutagenic effects onlyin the presence of S9 mix. Strain TA 1537 was reverted by thetriol but not by the diol. In the other bacterial strains thediol was more mutagenic than the triol, the difference in potencybeing largest in strain TA 100 (2.5-to 10-fold, depending onthe experimental conditions). In V79 cells, the diol was a potentmutagen, while the triol showed only very weak mutagenic effects.However the triol was more cytotoxic than the diol. High cytotoxicityof the triol was observed even in the absence of S9 mix. Theresults of the present study demonstrate that metabolites of3-OH-BP-7, 8-diol) are biologically-active derivatives of benzo[a]pyrene.Comparison of the mutagenic effectiveness in different bacterialstrains also reveals that metabolites of 3-OH-BP-7, 8-diol andof BP-7, 8-diol substantially differ in the kind of geneticalterations they evoke.