The challenges and possibilities of public access defibrillation.

Out-of-hospital cardiac arrest (OHCA) is a major health problem that affects approximately four hundred and thousand patients annually in the United States alone. It is a major challenge for the emergency medical system as decreased survival rates are directly proportional to the time delay from collapse to defibrillation. Historically, defibrillation has only been performed by physicians and in-hospital. With the development of automated external defibrillators (AEDs), rapid defibrillation by nonmedical professionals and subsequently by trained or untrained lay bystanders has become possible. Much hope has been put to the concept of Public Access Defibrillation with a massive dissemination of public available AEDs throughout most Western countries. Accordingly, current guidelines recommend that AEDs should be deployed in places with a high likelihood of OHCA. Despite these efforts, AED use is in most settings anecdotal with little effect on overall OHCA survival. The major reasons for low use of public AEDs are that most OHCAs take place outside high incidence sites of cardiac arrest and that most OHCAs take place in residential settings, currently defined as not suitable for Public Access Defibrillation. However, the use of new technology for identification and recruitment of lay bystanders and nearby AEDs to the scene of the cardiac arrest as well as new methods for strategic AED placement redefines and challenges the current concept and definitions of Public Access Defibrillation. Existing evidence of Public Access Defibrillation and knowledge gaps and future directions to improve outcomes for OHCA are discussed. In addition, a new definition of the different levels of Public Access Defibrillation is offered as well as new strategies for increasing AED use in the society.

[Skating fun and fractures of the maxillofacial bones]. / IJspret en fracturen van het aangezichtskelet.

Skating is a favourite sport in the Netherlands. Injury data on skating were collected from the emergency departments of 6 hospitals in the province of Friesland in the Netherlands during the Eleven Cities ice skating marathon over 200 kilometers in January 1997, with 16,688 participants. Among a total of 55 fractures, the maxillofacial skeleton had a relative high fracture frequency (n = 7; 13%), especially the zygomatic complex (n = 6); one patient had a mandibular fracture. An innocent-looking black eye, unilateral numbness of the upper lip and malocclusion following a skating accident merit full medical attention.

[Medical consequences of the 15th Eleven-Cities skating marathon]. / De medische gevolgen van de 15e Elfstedentocht.

The 15th Dutch Eleven Cities ice skating marathon was run on Saturday 4th January 1997. Numerous measures were taken to reduce the risk of accidents: active or standby were 7 hospitals, 3 helicopters for transport of casualties, 60 first-aid stations along the route, 600 first-aiders, 120 Red Cross workers, 50 GPs, 10 diving teams and a large police presence. A total of 150 ambulance rides were made and 151 skaters with 179 injuries were treated in the hospitals. The number of treatments administered in the first-aid stations was 2393 (14.3% of the total number of participants), including 829 cases of frostbite/hypothermia. Of the 16,387 participants of 1997, 11,523 (70.3%) finished.

[Eleven-Cities skating marathon and the windchill factor]. / Elfstedentocht en gevoelstemperatuur; 'windchill'-factor.

The weather conditions during the 15th Eleven Cities ice skating marathon (200 kilometers) in Friesland (the Netherlands) on 4 January, 1997, were better than predicted. For measurement of the windchill factor the Steadman scale is preferred to the Siple and Passel scale. For ice skaters the windchill factor is lower than for the spectators, due to the drag effect.

Genetic method for pre-classification of genotoxins into monofunctional or cross-linking agents.

To characterize environmental carcinogens, there is a need to distinguish monofunctional genotoxic agents from those having cross-linking potential, because chemicals which can cross-link DNA are among the most potent carcinogens in rodents [Barbin and Bartsch, 1989] and humans [Allen et al., 1988; Kaldor et al., 1988]. Here we provide a genetic method for a pre-classification of genotoxins with respect to their functionality--monofunctional versus cross-linking. The procedure is based on the determination of relative clastogenic efficiency by a two-endpoint comparison in Drosophila: (i) induction of chromosome loss (CL), (ii) incidence of recessive lethal mutations (RL). Analysis of CL/RL ratios of 53 genotoxins, all mutagens in Drosophila, permitted distinction of 45 into two major categories: (i) 21 monofunctional agents with CL/RL indices generally < or = 1; (ii) 24 agents with ratios > 2 exhibiting DNA cross-linking properties. Within the group of monofunctional agents, CL/RL ratios tend to be low for SN1 agents, i.e., for N-ethyl-N-nitrosourea, N-ethyl-N'-nitro-N-nitrosoguanidine, and for N-nitrosodiethylamine. With cross-linking agents, the number of reactive groups appeared of minor importance as bi-, tri-, and tetrafunctional agents showed no significant differences in their CL/RL indices. Among 8 chemicals which could not be grouped into one of the two categories are two (adriamycin, daunomycin) regarded as intercalating agents. It is concluded that this two-endpoint analysis in Drosophila has prognostic value and can assist in the characterization of genotoxic agents with unknown mode of action.

Validation of whole chick embryo cultures, whole rat embryo cultures and aggregating embryonic brain cell cultures using six pairs of coded compounds.

A comparative study was performed to assess the effects of six pairs of coded compounds using cultures of whole chick and rat embryos as well as aggregating brain cell cultures. Developed originally for basic studies in developmental biology, these three culture systems have been adapted for the screening of chemicals in the field of prenatal toxicology. Chick and rat embryos were cultured for 2 days during the early stages of organogenesis. Aggregating cell cultures were prepared from early foetal rat telecephalon and grown for 14 days in a chemically defined medium. Concentration-response relationships were established by treating whole embryos in vitro for 2 days, and aggregating brain cell cultures for 9 days. After decoding the compounds, the results showed that, in the three test systems, specific effects were induced at comparable concentration levels. Similar compound-related malformations could be observed in both chick and rat whole embryo cultures. In aggregating brain cell cultures, neuron- and glia-specific effects could be distinguished. Based on the results obtained in the three in vitro systems, the following concentration ranges were determined for the teratogenic/toxic potencies of the test compounds (in mol/litre): <10(-6): retinoids (Ro 13-6307, Ro 1-5488), 6-aminonicotinamide, ketoconazole; 10(-6)-10(-3): 4-hydroxypyridine, sulfadiazine, sulfanilamide, caffeine, theophylline, metronidazole, methoxyacetic acid; >10(-3): methoxyethanol. In general, the three in vitro test systems were found to provide concordant and complementary data on the toxicity and teratogenicity of a given compound. These data were also comparable with those available from in vivo studies. It is therefore concluded that such a test battery could contribute significantly to risk assessment and to the reduction of in vivo experimentation in reproductive toxicology.

Variation of spontaneous and induced mitotic recombination in different Drosophila populations: a pilot study on the effects of polyaromatic hydrocarbons in six newly constructed tester strains.

A set of six Drosophila strains was developed, by inducing by chemical treatment with N-ethyl-N-nitrosourea (ENU) new white and, in some strains, yellow mutations in 3 wild-type (WT) and 3 insecticide-resistant (IR) populations. These strains were previously shown to vary with regard to contents and inducibility of microsomal oxidative enzymes (Zijlstra et al., 1984). In this pilot study results from a first evaluation of these strains in somatic mutation experiments are reported, using as genotoxins an aromatic amine (2-naphthylamine, 2-NA), one substituted (9,10-dimethylanthracene, DA) and one non-substituted (benzo[a]pyrene, BP) polycyclic aromatic hydrocarbon. Developing larvae heterozygous for white were chronically exposed to three different exposure doses of each carcinogen. Adult females were inspected for the occurrence of mosaic light clones in their eyes, using the somatic mutation and recombination test (SMART). Evidence is presented indicating strong genotype-dependent variation in both spontaneous and chemically induced mutational and recombinational events in somatic cells of Drosophila. The spontaneous frequencies varied from 3.5% (Hikone-R), 4.3% (Berlin-K), 6.3% (Oregon-K), 9.1% (91-C), 20.5% (Haag-79) to 49.1% (91-R), corresponding to a 14-fold difference in spot frequencies between the two extremes. BP, DA and 2-NA were readily detectable in both Hikone-R (IR) and Oregon-K (WT), less so in 91-C (WT) and Haag-79 (IR), whereas the performance of strain Berlin-K (WT) was rather poor. The special problem with strain 91-R was the high frequency with which mosaic light spots occur not only in female genotypes heterozygous for white, but also in homozygous condition in the original stock. The up to 20-fold variation in induced spot frequencies between different genotypes poses questions for further investigations with respect to the genetic constitution of the various strains and the role of enzyme induction on somatic cell mutagenicity, which in this system is predominantly the result of mitotic recombination.

Importance of multiple hydroxylated metabolites in hexamethylphosphoramide (HMPA)-mediated mutagenesis in Drosophila melanogaster.

The mutagenic profiles in Drosophila and the influence of inhibition of metabolism on genotoxic activity were determined for hexamethylphosphoric triamide (HMPA), some synthetically prepared presumed metabolites and ethylated analogs. Demethylated HMPA metabolites are considerably less mutagenic than HMPA, dependent on the degree of demethylation. The mutagenicity of the presumptive primary metabolite, hydroxymethyl pentamethylphosphoramide (HM-Me5-PA), is comparable to HMPA and can be decreased considerably by inhibition of the metabolism by 1-phenylimidazole or iproniazid. This suggests that further oxidative metabolism is required for mutagenic activity. The mutagenicity of the doubly hydroxylated HMPA metabolite, N,N'-bis(hydroxymethyl)-tetramethylphosphoramide (N,N'-(HM)2-Me4-PA) can also be decreased by inhibition of metabolism, whereas the 3-fold hydroxylated N,N',-N"-(HM)3-Me3-PA is not affected by pretreatment with enzyme inhibitors, indicating that no further oxidative metabolism is required for its activation. A second hydroxylation on 1 dimethylamino group, forming N,N-(HM)2-Me4-PA, results in a drastic loss of mutagenic activity. Further oxidation of HM-Me5-PA to formyl pentamethylphosphoramide (formyl-Me5-PA) also leads to a strong reduction of the genotoxic activity. The rearrangement product of N-oxidation, N-[bis(dimethylamino)phosphinyl)-oxy)dimethylamine (HMPOA) is not mutagenic in Drosophila. The very low mutagenicity of hexaethylphosphoramide (Et6-PA) allowed us to study the mutagenicity of some ethyl-hydroxymethyl hybrid compounds. For the ethylated phosphoramides also the presence of only 1 hydroxymethyl group is insufficient for mutagenic activity, whereas the introduction of 2 or 3 hydroxymethyl groups resulted in considerable genotoxicity in the sex-linked recessive lethal (SLRL) test as well as in the ring-X loss test. It is concluded that the bioactivation of HMPA in Drosophila proceeds via multiple metabolic hydroxylations to form multifunctional, cross-linking agents. The presence of an oxygen atom on the phosphorus appears to be a prerequisite for the genotoxic activity of HMPA as hexamethylphosphorus triamide (HMPT), a derivative lacking this oxygen, is only weakly mutagenic in Drosophila. The results presented in this paper do not support the theory that formaldehyde is the active principle of activated HMPA.

Endometriosis: treatment with gonadotropin-releasing hormone agonist Buserelin.

Fifty-one women with pelvic endometriosis were treated with the gonadotropin-releasing hormone agonist (GnRHa) Buserelin (Hoechst Holland N.V., Amsterdam, The Netherlands) 300 micrograms three times a day intranasally for 6 months. Forty-nine women completed treatment; 42 were available for 6 months of follow-up following treatment. Symptoms showed prompt and significant improvement. Follow-up after treatment revealed persistent relief from dysmenorrhea and dyspareunia in, respectively, 58.6% and 88.2% of the women, whereas pelvic pain returned to pretreatment scores. Serum estradiol (E2) was suppressed to predominantly early follicular phase concentrations. Laparoscopy at the end of therapy showed significant reduction of scores for implants only. There was no relation between the degree of E2 suppression during therapy and the improvement of symptoms or the reduction of endometriosis. Statistical analysis in 22 infertile patients, of whom 7 conceived during follow-up, revealed no differences in E2 levels during therapy, improvement of symptoms, or reduction of endometriosis. Buserelin appears to be safe, well tolerated, and effective in the management of endometriosis and associated complaints.

Liquid holding increases mutation induction by formaldehyde and some other cross-linking agents in Escherichia coli K12.

The induction by some cross-linking agents of forward mutations leading to nalidixic acid resistance in Escherichia coli K12/343/113 was considerably enhanced when a 24-h period of liquid holding was interpolated between treatment and growth phase. Liquid holding increased the mutagenic effectiveness of nor-nitrogen mustard (NNM) 28-fold, of phosphoramide mustard (PAM) 10-fold, and of tris-ethyleneimino)-phosphineoxide (TEPA), tris(chloroethyl)amine (TCEA) and chloroacetaldehyde (CAA) 3-fold, over the complete concentration range. By contrast, the activities of cisplatin (CDDP), transplatin (TDDP) and chloracetamide-N-metholol (CAM) were slightly decreased after liquid holding. Liquid holding did not measurably influence the mutagenicity of formaldehyde at low concentrations, whereas at higher concentrations an 8-fold increase was observed. As opposed to the considerable activity in the Uvr+ strain, formaldehyde was found not to be mutagenically active in an E. coli strain carrying a deletion of the uvrB gene.

Influence of metabolic factors on the mutagenic effectiveness of cyclophosphamide in Drosophila melanogaster.

This paper describes the influence of changes in metabolic activity on the in-vivo mutagenic effectiveness of cyclophosphamide in Drosophila melanogaster. A dose-dependent increase in mutagenicity was observed until a plateau value is reached which was increased only slightly after enzyme induction with Aroclor 1254, whereas induction with phenobarbital resulted in a decrease, especially when cyclophosphamide was applied by injection. Treatment of the adult males with inhibitors of the monoamine oxidase (MAO, EC 1.4.3.4), such as iproniazid (Ipr), benzimidazole or tryptamine, led to a marked increase of the mutagenic effectiveness of cyclophosphamide especially in spermatocytes. This indicates the importance of metabolic de-activation processes for the limited mutagenicity of cyclophosphamide in Drosophila. The principal active metabolite of cyclophosphamide, phosphoramide mustard, is extensively de-activated by enzymes that can be inhibited by 1-phenylimidazole (PhI), presumably cytochrome P-450 (EC 1.14.14.1), but not by those blocked by MAO inhibitors. Inhibition of the FAD-containing dimethylaniline monooxygenase (FDMAM, EC 1.14.13.8) by N,N-dimethylbenzylamine (N,N-DMB) resulted in some increase in cyclophosphamide mutagenicity only in spermatids. The marginal mutagenicity of cyclophosphamide in Drosophila larvae could not be increased either by cytochrome P-450 induction with phenobarbital or by MAO inhibition with Ipr. In contrast to the failure of cyclophosphamide to induce rod-chromosome loss, a considerable activity was found when a ring-shaped chromosome was used. Similar to the sex-linked recessive lethal (SLRL) test, ring-X loss frequency could be enhanced by simultaneous treatment with MAO inhibitors. The observed ring-X loss frequency declined when males treated with cyclophosphamide were mated to DNA-repair deficient mei-9L1 females. Cyclophosphamide produces chromosome breaks, detected as 2-3 translocations, in Drosophila spermatocytes, the stage in spermatogenesis that is also the most sensitive to the induction of SLRL mutations.

Influence of inhibition of the metabolic activation on the mutagenicity of some nitrosamines, triazenes, hydrazines and seniciphylline in Drosophila melanogaster.

It is determined to what extent certain inhibitors of the xenobiotic metabolizing enzyme systems have an influence on the mutagenicity of various pro-mutagens in Drosophila. 1-Phenylimidazole (PhI) is used as an inhibitor of the cytochrome P-450 (P-450) mediated monooxygenase activities. Iproniazid (Ipr) is a typical monoamine oxidase (MAO) inhibitor which as well seems capable of inhibiting to a certain extent P-450 mediated metabolism. N, N-Dimethyl benzylamine (N, N-DMB) is used as a competitive substrate for the N-oxidizing flavin-containing dimethylaniline monooxygenase (FDMAM). The enzyme-inhibiting activities of PhI and Ipr were determined in vitro using microsomes obtained from Drosophila larvae and adults. Both compounds were capable of inhibiting benzo[a]pyrene (BP) hydroxylation and p-nitroanisole (p-NA) demethylation, although for Ipr 100-fold higher concentrations were required compared to PhI. As model-mutagens were used: the nitrosamines dimethylnitrosamine (DMN) and diethylnitrosamine (DEN), the triazenes 1-(2,4,6-trichlorophenyl)-3,3-dimethyltriazene (Cl3PDMT), 1-(3-pyridyl)-3,3-dimethyltriazene (PyDMT) and dacarbazine (DTIC), the hydrazines procarbazine (PCZ), 1,1-dimethylhydrazine (1,1-DMH) and 1,2-dimethylhydrazine (1,2-DMH) as well as the pyrrolizidine alkaloid seniciphylline (SPh). Simultaneous or pretreatment with Ipr results in a clear decrease of the mutagenicity of Cl3PDMT, while PhI pretreatment leads to an increased mutagenicity. This indicates that these two inhibitors do not inhibit the same enzyme or isozyme. For SPh too, Ipr pretreatment results in some decrease of the mutagenicity. This is in contrast to DEN, where the activation is clearly inhibited by PhI while Ipr has only a minor effect. For DMN, DTIC and PCZ both Ipr and PhI pretreatment caused considerable decreases of the mutagenicity. Inhibition of the FDMAM catalyzed activity by N,N-DMB resulted in an increase of mutagenicity with Cl3PDMT, in a moderate decrease of mutagenicity with DTIC, and a marked decrease with DMN, which was strongly inhibited. In contrast to the clear-cut mutagenicity of PCZ, 1,1-DMH and 1,2-DMH are not mutagenic in Drosophila. No change was observed upon inhibition of the various metabolizing activities. Apart from using strain differences in metabolizing activities and enzyme induction, enzyme inhibition can also be used to determine the influence of metabolism on the in vivo mutagenicity of promutagens in Drosophila.

The ratio of induced recessive lethals to ring-X loss has prognostic value in terms of functionality of chemical mutagens in Drosophila melanogaster.

For 25 mutagens in Drosophila the ratio was determined between the induction of sex-linked recessive lethals (SLRL) and the induction of ring-X loss in male adults. For small monofunctional alkylating agents this ratio increases with decreasing s-value from 1.8 for methyl methanesulfonate (MMS) to 27 for ethylnitrosourea (ENU). For multifunctional cross-linking agents, however, the ratio varies within relatively narrow limits, ranging from 0.15 for cisplatin to 0.07 for tris-(1-aziridinyl)phosphineoxide (TEPA), while for most agents the ratio is around 0.12. The number of reactive groups seems to be of minor importance for compounds with more than one functionality as bi- and tri-functional agents show similar ratios. The systemic difference in the ratios between mono- and multi-functional agents suggests that different mechanisms are involved in the induction of SLRLs and ring-X loss. For ethyleneimine (EI) and ethyleneoxide (EO) low ratios of 0.32 and 0.60 respectively were observed which do not correlate with their s-values. An alternative chromosome-breaking mechanism may be responsible for this deviation, possibly alkylation of the phosphate backbone of DNA, followed by an intramolecular displacement of one of the deoxyribose groups by the beta-amino or the beta-hydroxy group. It is felt that the considerable difference between the ratios for monofunctional and multifunctional agents may be of prognostic value and can be used to obtain information on the mechanisms of mutagens with 'unknown' action, provided that structural features are taken into account. Hexamethylphosphoramide (HMPA), hexamethylmelamine (HEMEL), tetramethylurea (TMU) and dimethylpropyleneurea (DMPU) all show SLRL: ring-X loss ratios that match those of multifunctional agents, 0.08, 0.12, 0.08, and 0.16, respectively. The ratios for the pyrrolizidine alkaloids monocrotalin and seniciphilline, 0.053 and 0.24 respectively, also correspond with this group of mutagens. The low ratios for formaldehyde, 2-chloro-acetaldehyde and 2-chloroethyl methanesulfonate, 0.30, 0.052 and 0.36 respectively, are indicative that cross-linking may attribute considerably to their mutagenic action in Drosophila. On the other hand, not all mutagens containing 2 reactive groups act as cross-linking agents. The ratio for 1,2-dibromoethane, 2.7, indicates that it may act as a monofunctional agent. This is in accordance with the proposed activation mechanism by glutathione S-transferase, producing a monofunctional half-mustard derivative (Rannug, 1980; van Bladeren et al., 1981).

Metabolic inactivation of mutagens in Drosophila melanogaster.

The route of administration of a drug is a pharmacological factor to be reckoned with. In Drosophila, a whole-animal object for mutagenicity studies, the way in which a mutagen is applied can also be of crucial importance. In this study the mutagenicity of a number of directly acting agents was determined after feeding or injection of the mutagen. Methyl-p-toluenesulphonate (Me-Tos), ethyl-p-toluenesulphonate (Et-Tos) and nor-nitrogen mustard (NNM) were not mutagenic in a sex-linked recessive lethal test when fed to the adult flies. Injection, however, did produce significant mutagenicity. The absence of mutagenicity after oral application is not caused by chemical instability but is the result of metabolic de-activation, presumably in the gut and the fat body. Feeding of these compounds in combination with the inhibition of cytochrome P-450 by 1-phenylimidazole (PhI) allowed sufficient quantities of the mutagen to reach the gonads and to produce significant genetic damage. This resembles what is known in pharmacology as a 'first-pass effect'. Formaldehyde (FA) mutagenicity, which also is only observed after injection and not in feeding experiments, was not affected by either iproniazid (Ipr) or PhI pretreatment. Aspecific enhancement of mutagenicity is excluded as this effect was not observed with mutagens that are structurally related to the tosylates, such as methyl methanesulphonate (MMS), ethyl methanesulphonate (EMS) or hycanthone methanesulphonate (HyMS). A number of other inhibitors of metabolism did not influence metabolic de-activation in Drosophila.

Somatic cell mutagenicity in Drosophila melanogaster in comparison with genetic damage in early germ-cell stages.

With the intention of assessing the general performance, sensitivity and the underlying mechanisms of somatic cell mutagenicity assays in Drosophila, a study was undertaken to compare the effectiveness of 5 procarcinogens and 4 direct-acting agents in the white/white-coral eye mosaic assay (SMART) with their activity in early (premeiotic) male and female germ-cell stages, after exposure of Drosophila larvae. The outcome indicated a lack of agreement in the results from recessive lethal assays (SLRL) in comparison with the somatic mutation and recombination test (SMART). The procarcinogens 2-naphthylamine (NA), 3-methylcholanthrene (MC), 9,10-dimethylanthracene (DA) and 7,12-dimethylbenz[a]anthracene (DMBA), and the direct-acting mutagens bleomycin (BM), methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS), were quite efficient in producing somatic recombination and mutations in white/white-coral larvae, as opposed to only weak effects in early germ-cell stages. 2-Acetylaminofluorene (2AAF) showed marginal effects in both germ cells and somatic tissue after exposure of female larvae, but was inactive in testis. The discrepancy in mutational response between somatic cells and premeiotic germ cells is most impressive for MMS and BM. There is sufficient evidence for attributing a good sized proportion of the encountered variation to efficient error-free DNA repair of premutational damage and to segregational elimination during meiosis of deleterious mutations: (1) The efficient point mutagen ENU was the but one agent producing high levels of viable genetic alterations in early germ cells and in somatic cells. A similar behaviour was previously described for diethylnitrosamine, which ethylates DNA in the same fashion as ENU. (2) In early germ-cell stages of mei-9L1 male larvae, MMS induced multiple mutations (putative clusters) at a low dose differing by a factor 20-40 from those needed to produce an equivalent response in repair-competent strains. This is consistent with the concept of an active excision repair in premeiotic cells. (3) In the case of EMS, next to DNA repair, germinal selection seems to restrict the realization of EMS-induced genetic damage in premeiotic cells. (4) Bleomycin-induced chromosome aberrations caused high mortality rates in males (hemizygous for an X-chromosome) but not in females. MMS and BM, agents known to show preference for chromosome aberration induction, produced 3-6-fold higher rates of somatic mutational events (SME) in female genotypes as compared with the other sex.(ABSTRACT TRUNCATED AT 400 WORDS)

Mechanistic and methodological aspects of chemically-induced somatic mutation and recombination in Drosophila melanogaster.

This study presents the analysis of chemically-induced somatic mutations and chromosomal damage in the eye imaginal discs of Drosophila larvae, assayed later as twin (TS) and single light (LS) mosaic spots in the adult eyes. Regarding the question as to what kind of DNA alterations contribute to somatic cell mutagenicity, the approach followed here has been to investigate the possible differences in response between male (hemizygous for an X) and female (homozygous) larvae, rod-X/rod-X versus ring-X/rod-X genotypes and inversion-heterozygotes versus genotypes not carrying an inversion. The systems chosen for this analysis were the white-coral/white (wco/w) and the white+/white (w+/w) eye mosaic system. The principle findings with 12 mutagens of different modes of action are as follows: (1) At least 98% of all TS and LS induced by cisplatin (DDP) in wco/w female larvae and about 95% of those by formaldehyde (FA) appear as the result of recombinogenic activity between the two homologous X-chromosomes. The corresponding estimates for MMS, EMS and ENU are 81%, 73% and 61%, respectively. (2) The long scS1L sc8R inversion, which also contains In(1)dl-49, suppresses induction of TS to 83-93%. There was also a sharp decline in the frequency of LS in inversion heterozygotes for DDP (91%), FA (86%), MMS (52%) and EMS (47%). (3) Ethylnitrosourea (ENU) was the mutagen for which introduction of the inverted chromosome reduced only slightly (23%) the frequency of LS, indicating that the majority of them were somatic mutations (and deletions) at the white locus. (4) In w/RX females heterozygous for a ring-X chromosome, the frequency of LS was only approximately one tenth of that of the control (w+/w) group, after exposure to MMS or DDP. The explanation is that exchange processes involving the ring frequently lead to genetic imbalance with subsequent cell killing.

The effect of mixed-function oxidase and amine oxidase inhibitors on the activation of dialkylnitrosamines and 1,2-dimethylhydrazine to bacterial mutagens in mice.

The effect of the mixed-function oxidase inhibitor phenylimidazole (PI) and the amine oxidase inhibitors iproniazid (IPRO) and aminoacetonitrile (AAN) on the mutagenic activity of various carcinogens was determined in intrasanguineous host-mediated assays, using mice as hosts and E. coli 343/113 as an indicator of mutagenic activity. The carcinogenic compounds dimethyl-, diethyl-, methylethyl-, and diethanolnitrosamine (DMNA, DENA, MENA, and DELNA respectively) and 1,2-dimethylhydrazine (SDMH) were administered i.p. to mice pretreated or not with one of the inhibitors. After 4 h exposure to each of the carcinogens, E. coli cells recovered from the liver of non-pretreated mice showed considerable induction of VALr mutations; after pretreatment of the hosts with the three inhibitors, significant reduction of the amounts of induced mutants in vivo was observed. Particularly, PI proved a very efficient inhibitor of DENA, MENA, DELNA, and SDMH mutagenicity (93%-97% reduction), suggesting that these carcinogens are mainly activated by cytochrome P-450-dependent enzymes. However, since PI might also inhibit the NAD-mediated activation of DELNA by alcohol dehydrogenase (ADH), the present experiments do not rule out an additional role of ADH in the in vivo mutagenic activation of DELNA. AAN and IPRO were less and much less effective, respectively, in reducing the mutagenic activity of all compounds. Surprisingly, PI showed less inhibition of the mutagenic activity of DMNA (60% reduction), as compared to the other carcinogens; this indicates that metabolic routes other than the cytochrome P-450-dependent enzyme system may be important for the activation of DMNA.

Analysis of hexamethylphosphoramide (HMPA)-induced genetic alterations in relation to DNA damage and DNA repair in Drosophila melanogaster.

This paper reports the results of a study on the mutagenic profile of HMPA in Drosophila melanogaster. HMPA produced all types of genetic damage tested for in post-meiotic cells of treated males; at the concentrations used, recessive lethals and ring-X losses were induced at significant rates while 2-3 translocations, entire and partial Y-chromosome losses only occurred at low rates. From a comparison with alkylation-induced mutational spectra, we note a number of peculiarities of HMPA mutagenesis: there is no storage effect on HMPA-induced translocations; the ratio of F2-lethals: F3-lethals varies from 6:1 to 9:1, indicating a low capacity of HMPA for delayed mutations; the use of the DNA-repair-deficient mei-9L1 females instead of an excision-proficient control strain has no influence on the recovery of mutations (recessive lethals) induced in males; the high frequencies of chromosome loss (CL) induced by HMPA, which are mostly due to ring-X loss, leads us to speculate that one (or more) of its metabolites acts as a DNA-crosslinking agent. In experiments on maternal effects with mei-9LI females, there is a 20-40% reduction in the rates of induced CL. Conversely, with mei-41D5 females, there is a weak increase in CL frequencies. HPLC analysis of DNA reacted with [14C]HMPA exhibits no methylation at the O6 or the N-7 of guanine. This finding, together with the observed inactivity of hexaethylphosphoramide (HEPA) in the recessive lethal assay, suggests that the formation of DNA-bound forms from HMPA may not be the result of simple methylation reactions. This conclusion is supported by the genetic data, i.e., the lack of a storage effect on HMPA-induced chromosome rearrangements. Consistent with a hypothesis by Brodberg et al. (1983) to explain the action of cisplatin in Drosophila, comparisons of the spectra of genetic alterations produced by HMPA, A 139 (bifunctional) and Thio-TEPA (trifunctional) in the assay for chromosome loss suggest the involvement of two distinct mechanisms in the formation of ring-X loss by crosslinking agents. One pathway concerns induction of chromosome loss as a consequence of sister-chromatid exchanges (SCEs). The second mechanism may be due to DNA adducts or a single adduct responsible for both a fraction of CL and for induced partial Y-loss (PL). Inactivation of the mei-9+ function has two consequences: SCE-mediated ring-X loss frequency is lowered in mei-9 females in comparison to the repair-proficient control strain, while the opposite effect is indicated for that fraction of ring-X loss generated by the second mutational pathway.(ABSTRACT TRUNCATED AT 400 WORDS)