Flumequine enhances the in vivo mutagenicity of MeIQx in the mouse liver.

The combined effects of various carcinogens found in food products are a concern for human health. In the present study, the effects of flumequine (FL) on the in vivo mutagenicity of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in the liver were investigated. Additionally, we attempted to clarify the underlying mechanisms through comprehensive gene analysis using a cDNA microarray. Male gpt delta mice were fed a diet of 0.03 % MeIQx, 0.4 % FL, or 0.03 % MeIQx + 0.4 % FL for 13 weeks. The effects of cotreatment with phenobarbital (PB) were also examined. Treatment with MeIQx alone increased gpt and Spi(-) mutant frequencies, and cotreatment with FL, but not with PB, further exacerbated these effects, despite the lack of in vivo genotoxicity in mice treated with FL alone. FL caused an increase in Cyp1a2 mRNA levels and a decrease in Ugt1b1 mRNA levels, suggesting that the enhancing effects of FL may be due in part to modification of MeIQx metabolism by FL. Moreover, FL induced an increase in hepatocyte proliferation accompanied by hepatocellular injury. Increases in the mRNA levels of genes encoding cytokines derived from Kupffer cells, such as Il1b and Tnf, and cell cycle-related genes, such as Ccnd1 and Ccne1, suggested that FL treatment increases compensatory cell proliferation. Thus, the present study clearly demonstrated the combined effects of 2 different types of carcinogens known as contaminants in foods.

The effects of long-term exposure to ozokerite mainly consisting of an aliphatic series of hydrocarbons using F344 rats.

Combined chronic toxicity and carcinogenicity studies of ozokerite (OZK), a natural wax substance used as a food additive for a gum base, were performed in male and female F344 rats. Dietary concentrations of 0%, 0.05%, 0.1% and 0.2% OZK were applied in a 52-week chronic toxicity study and 0%, 0.1% and 0.2% in a 104-week carcinogenicity study. In the chronic toxicity study, treatment with OZK caused a xenobiotic reaction against absorbed OZK, including formation of histiocytosis and granulomas with crystalline material in many organs in all of the treated males and females. Particularly in the liver, granulomatous inflammation was accompanied by hepatocellular vacuolation and changes in the serum biochemical parameters indicative of hepatic disorder. The number and area of glutathione S-transferase placental form (GST-P) positive foci were increased in all of the treated groups of both sexes, suggesting the proliferative effect of OZK. In the carcinogenicity study, the incidence of hepatocellular adenoma and the total tumor incidence in the liver of all of the treated males were significantly increased compared with the controls. In conclusion, long-term exposure to OZK caused systemic chronic inflammation due to a foreign body response. OZK was weakly carcinogenic in the liver of male F344 rats.

Comprehensive toxicity study of safrole using a medium-term animal model with gpt delta rats.

In order to investigate a medium-term animal model using reporter gene transgenic rodents in which general toxicity, genotoxicity and carcinogenicity are evaluated, F344 gpt delta rats were given a diet containing 0.1% and 0.5% (a carcinogenic dose) safrole for 13 weeks. Serum biochemistry and histopathological examinations revealed overt hepatotoxicity of safrole, in line with previous reports. In the current study, safrole treatment possibly resulted in renal toxicity in male rats. In the in vivo mutation assays, an increase or a tendency to increase of the gpt mutant frequencies (MFs) was observed in both sexes at the carcinogenic dose. The number and area of foci of glutathione S-transferase placental form (GST-P) positive hepatocytes, ratio of proliferating cell nuclear antigen (PCNA)-positive hepatocytes and 8-hydroxydeoxyguanosine (8-OHdG) levels in liver DNA were significantly increased in both sexes of the 0.5% group. The overall data suggested that the present model might be a promising candidate for investigating comprehensive toxicities of the agents. In addition, data demonstrating the base modification and cell proliferation due to exposure to safrole could contribute to understanding safrole-induced hepatocarcinogenesis, which imply expanding in application of this model.

Investigation of carcinogenicity for levamisole administered in the diet to F344 rats.

A two year carcinogenicity study of anthelmintic drug levamisole (LV) was performed using 50 male and 50 female F344 rats at dietary drug concentrations of 0, 60, or 300 ppm. The daily intakes of LV were calculated to be 2.6, 12.9 mg/kg b.w./day for males and 2.9, 14.1mg/kg b.w./day for females, respectively. No significant differences in general condition and survival rate (82%, 74%, 80% in males and 84%, 84%, 84% in females, respectively) were observed. In the 300 ppm group, suppression of body weight gain was observed from the onset of treatment and reduction in final body weights was 6% in males and 11% in females. Significant increases in the absolute and/or relative weights of the lungs, heart, spleen, liver, kidneys, and adrenals were observed in males and/or females treated with 300 ppm. Some of high incidences neoplasms were observed, and there were also tendencies to increase for mammary gland fibroma and thoracic/abdominal cavity mesothelioma in males. However, there were no significant inter-group differences in incidences, histopathological types or differences compared with historical control data. Thus, it was concluded that LV was not carcinogenic to male and female F344 rats under the experimental conditions.

[Antimicrobial activity of clarithromycin and its effect on bacterial adherence to medical material].

Antimicrobial activity of clarithromycin (CAM) in comparison with other 6 macrolides was determined against 419 recent clinical isolates including Streptococci, Enterococci, Staphylococci, Moraxella, Haemophilus and Bacteroides strains. MICs80's of CAM against Streptococcus pneumoniae, Streptococcus pyogenes and Streptococcus agalactiae were 0.78, 0.10 and 0.10 microgram/ml, respectively. MICs80's against Moraxella catarrhalis and Haemophilus influenzae were 0.05 and 1.56 micrograms/ml, respectively. Laboratory-induced resistance to CAM in strains of S. pneumoniae, Enterococcus faecalis or methicillin-sensitive Staphylococcus epidermidis occurred in stepwise fashion and at a very low rate. Adherence to silicon filter of strains of Pseudomonas aeruginosa or S. epidermidis was strongly repressed by the addition of sub-MIC of CAM or other macrolides. Although tosufloxacin (TFLX) alone had no bactericidal activity against the sessile cells of P. aeruginosa strains tested, TFLX showed synergistic bactericidal activity when combined with sub-MICs of CAM or erythromycin.

Genetic analysis of population structure in marine teleosts around Japan.

Genetic differentiation and genetic variability were estimated from data on electrophoretically detectable isozymic loci in 12 species of marine teleosts belonging to 10 families of six orders, this study permitted conclusions concerning population structures within the species of marine teleosts. As a result the 12 species could be grouped into two classes, one of which showed higher genetic diversity and included the species inhabiting coastal areas; the other showed lower genetic diversity and included the species inhabiting offshore areas and spawning free pelagic eggs. Class I and Class II could be further divided into two groups (I-1 and I-2, and II-1 and II-2) on the basis of the level of average heterozygosity. Based on the above classification and ecological characteristics, the degree of genetic differentiation may be defined mainly by the level of mixing of eggs and by fry migration, and only to a small extent by adult migration between localities. The degree of average heterozygosity should be a measure of effective population size of the breeding units within the species but not of the whole species. Putting together the above facts, six types of population structure have been proposed for marine teleosts as follows: Species in type I-1 have formed some large populations with large breeding units in offshore areas; Type I-2-1, species have formed a large number of breeding populations with relatively large effective sizes of breeding units in the coastal areas or rivers where they were hatched, even though the species may have migrated over a wide pelagic area during their juvenile to adult stages. The species in type I-2-2 would have formed a number of local subpopulations with large to intermediate breeding units in the coastal areas they inhabit. Type I-2-3 species have formed two or more completely isolated local races (or subspecies) with relatively large breeding units, but with a low level of genetic differentiation within each local race. The species in Type II-1 would have formed a single large population consisting of a few breeding units with large effective sizes in offshore areas around Japan. Type II-2 species would have formed one large population consisting of a large number of breeding units with small effective sizes in wide offshore areas around Japan.

Electromechanical changes during electrically induced and maximal voluntary contractions: surface and intramuscular EMG responses during sustained maximal voluntary contraction.

Changes in the electrical activity of the human gastrocnemius and soleus muscles during fatiguing maximal plantar flexions were studied with computer-aided EMG frequency power spectral analysis and intramuscular spike amplitude-frequency histogram analysis. In some experiments, brief supramaximal nerve stimulations of 80 Hz were given at 15-s intervals during sustained maximal voluntary contractions (MVCs). Multiple muscle biopsy samples were also obtained from the gastrocnemius muscle for fiber type determination. The surface EMG frequency spectral analysis showed a highly significant reduction in mean power frequency and root mean square EMG amplitude during sustained MVCs. The intramuscular spike amplitude-frequency histograms showed that the gastrocnemius muscle had a progressive reduction in the motor unit discharge frequency, particularly those with a relatively high amplitude, whereas the soleus muscle hardly showed a reduction in motor unit activity. Reduction in motor unit activity was also found to be more pronounced in gastrocnemius muscles with higher proportions of type II fibers. Brief maximal tetanic stimulations initially matching the MVC failed to increase the contraction force. Similarly, the evoked compound mass action potentials showed little change in the amplitude in subjects with different muscle fiber compositions. Results of this study suggest that during sustained MVCs, force fatigue could not be attributed to a failure of muscle membrane electrical propagation; a progressive reduction in motor unit activation does not result in a functional disadvantage, but may optimize excitation-contraction coupling by avoiding a muscle electrical conduction failure; and the extent of the reduction in motor unit activation seems to be muscle-fiber-type-dependent which may account for the reduction in amplitude and frequency of the surface EMG.

Electromechanical changes during electrically induced and maximal voluntary contractions: electrophysiologic responses of different muscle fiber types during stimulated contractions.

The electrophysiologic and mechanical responses of a synergistic muscle group composed of different muscle fiber types were studied with respect to excitation frequency and muscle fatigue in five men. The force and evoked action potentials obtained from surface and intramuscular fine wire electrodes were recorded continuously during stimulated contractions of the gastrocnemius and soleus at 20, 50, and 80 Hz. The stimulus voltage was adjusted so that the force generated by high-frequency tetani (50 and 80 Hz) could initially match the force of maximal voluntary contractions (MVCs). The surface and intramuscular EMG signals were digitized at a sampling rate of 10 kHz with 16-bit fast A/D converters and stored on a floppy disk. The digitized data were then processed for every 5 s to calculate evoked potential amplitude and conduction time using an HP 9836 computer. Results indicated that after 30 s of high-frequency stimulation, significantly less force was generated than after a similar period of MVC. During this period of high-frequency force fatigue, considerably greater force was developed at 20-Hz stimulation. The excessive force loss during high-frequency tetanic contractions was accompanied by a marked reduction in the evoked potential amplitude and conduction time (prolongation of the M wave). The recording of intramuscular evoked potentials showed the gastrocnemius muscle to have greater reductions in these parameters. Our results support the hypothesis that force fatigue during high-frequency stimulation results from failure of electrical propagation due to reduced muscle membrane excitability. The observed muscle-fiber-dependent electrophysiologic responses may suggest that the metabolic profile of muscle fibers plays an important role in regulating the muscle membrane excitability during high-frequency stimulation.