Recently, national guidelines have advocated greater use of atypical rather than typical antipsychotics in the treatment of schizophrenia. In addition, there have been safety concerns regarding the potential cardiotoxicity of certain antipsychotics taken in overdose. This has led regulatory authorities in the United Kingdom to restrict the use of thioridazine. The overall impact of these legislative changes on patterns of antipsychotic prescribing has received comparatively little attention. Therefore, we sought to examine the effects on community prescribing practices, and to determine whether this was accompanied by changes in patterns of antipsychotic poisoning. Between 2000-03, there was a rapid decline in the use of typical antipsychotics, whereas the use of atypical antipsychotics increased. The prevalence of atypical and typical antipsychotic prescribing has been approximately equal between 2003-06. During the same study period, hospital admissions due to typical antipsychotic poisoning also declined, however, the effects lagged behind changes in prescribing practice by 2-3 years. These data indicate that legislative changes that restrict the use of thioridazine and other typical antipsychotics are associated with a measurable reduction in the number of hospital admissions due to overdose with these agents.
It has been noted that chemical-induced initial insult is sometimes no longer detected in examinations after additional consecutive treatments, suggesting that the target organs acquire resistance to the chemical toxicity. In this study, whether acquired resistance to the skeletal muscle toxicity is observed during repeated treatment of a toxic dose of Compound A that has a 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitory activity was examined. F344 male rats (7-weeks old) were given a mixed diet with 0.12% Compound A (corresponding to approximately 100 mg/kg/day) for up to 56 days. Blood samples were obtained from the tail vein periodically during the dosing period, and utilized for the measurement of creatine kinase (CK) as a marker of skeletal muscle injury. In the necropsies on Days 4, 8, 11, 28, 42 and 56, the skeletal muscles from the rectus femoris were removed for histopathology or gene expression analysis. A satellite group was provided to measure the plasma concentrations of Compound A and M1, the active metabolite of Compound A. CK levels increased from Day 9 and reached approximately 30 times those of the controls on Day 12. Histopathology of the skeletal muscle on Day 11 revealed severe necrosis of the muscle fibers. However, in spite of continuous treatments to the damaged rats, the CK levels decreased after that and returned to normal levels on Day 18. No skeletal muscle injury was observed on Days 42 and 56. There were no marked differences in the exposure levels of Compound A and M1 between Days 8 (prior to CK elevation) and 28 (post CK elevation). As for the most significant changes in the gene expression analysis for the skeletal muscle on Days 42 and 56, the probe for IκBa, which is known as an inhibitor for nuclear factor-κB (NF-κB), increased 2-fold compared to the control. Furthermore, an increased probe for CCAAT/enhancer-binding protein (C/EBP) delta, a transcriptional factor, and a decreased probe for cAMP-response element-binding protein (CBP)/p300, a transcriptional coactivator, were also noted significantly on Day 56. These changes in the gene expression analysis suggested suppressed NF-κB-mediated transactivation, which was responsible for the protective effects on the muscle injury. Based on the present findings, the resistance to skeletal muscle injury observed in this study may be attributable to the suppressed NF-κB-mediated transactivation, but not to the decreased exposure to toxicants.
Toxicogenomics is a promising new tool for prediction of chemical toxicities including carcinogenicity in a relatively short period. However, it is important to develop a reliable animal test protocol for toxicogenomics studies. The preparation of RNA and tissues is also crucial, since it greatly influences outcomes of gene expression analysis. In the present study, we examined an animal test protocol by comparing gene expression data from different conditions and proposed a reliable animal test protocol for toxicogenomic studies. With regard to the preparation of tissues and RNA, here we present evidence that quality of RNA and tissues is well-preserved even after freezer storage for up to 2.5 years. Gene expression levels were compared using a GeneChip System (RGU34A, Affymetrix, Santa Clara, CA, USA) between RNA samples that were freshly prepared, stored at −80°C or re-prepared from tissue kept at −20°C. None showed degradation and no significant differences in expression were evident among the three sets of samples. The data demonstrate that gene expression analysis by DNA microarray is suitable for RNA or tissues that have been stored at an appropriate temperature.
Toxicogenomics is a promising new tool for prediction of chemical toxicities including carcinogenicity in a relatively short period. However, it is important to develop a reliable animal test protocol for toxicogenomics studies. The preparation of RNA and tissues is also crucial, since it greatly influences outcomes of gene expression analysis. We proposed an animal test protocol for toxicogenomics studies. In the present study, we examined an animal test protocol by comparing biological and gene expression data from different laboratories running identical in vivo studies on the same microarray platform. The results gave good correspondence in all three laboratories at the level of biological responses and gene expression, especially for genes whose expression changes were quite large. As the fold change or the signal values become smaller, however, discrepancies occur in gene expression data. For example, one laboratory shows an opposite directional change to the other two or no change. The results of hierarchical clustering and principal component analysis (PCA) demonstrated all samples from the three laboratories being clearly divided between control and treatment. Examination of the reproducibility of gene expression data across laboratories using the proposed animal test protocol thus confirmed only minor differences, which was expected to present no problems for gene expression analysis.
Dose- and time-dependent effects of 2,3,7,8-tetrabromodibenzo-p-dioxin (TBDD) on the liver were examined by single administration of TBDD by gavage to male and female rats. Fifteen Wistar rats of each sex per group received 0, 10, 30, 100 or 300 μg TBDD/kg body weight. Rats surviving to scheduled necropsy on Day 2, 7 or 36 after the TBDD administration were examined for hepatic histopathology, activities of hepatic microsomal enzymes and serum levels of lipids, total cholesterol and transaminases and hepatic concentrations of TBDD. Tigroid basophilic cytoplasm and hepatocellular hypertrophy were observed at 10 μg/kg on Day 2 or 7 through 36, whereas degenerative and aggressive lesions such as necrosis, fibrosis, multinucleated hepatocytes and disarrangement of hepatocytes occurred later at higher dose levels. Persistently increased activities of hepatic aryl hydrocarbon hydroxylase (AHH), ethoxycoumarin O-deethylase (ECOD) and ethoxyresorufin O-deethylase (EROD), increased serum levels of total cholesterol and phospholipid and increased relative liver weight were observed in all groups dosed 10 μg/kg and above, suggesting that hepatic microsomal monooxygenases and basophilic cytoplasm of hepatocytes were early and sensitive indicators among those TBDD-induced effects. A dose-dependent increase in liver concentrations of TBDD on Day 2 was followed by logarithmic decreases in TBDD concentrations against the days elapsed after the TBDD administration. An elimination half-life (t1/2) of TBDD from the liver was estimated to range from 12 to 16 days. It was suggested that females were more susceptible to TBDD than males, and that acute hepatotoxicity of TBDD was as potent as that of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).
Inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase occasionally cause myopathy characterized by weakness, pain, and elevated serum creatine phosphokinase (CK). In this study, we investigated the effects of simvastatin, an HMG-CoA reductase inhibitor, on the viability and insulin-like growth factor-1 (IGF-1) signaling in differentiating C2C12 mouse myoblast cells. Simvastatin decreased cell viability and CK activity, a marker of myogenesis, in differentiating cells in a dose-dependent manner. Although the simvastatin-induced decrease in viability in proliferating and differentiated cells was completely abolished by mevalonate or geranylgeranyl-pyrophosphate, the inhibitory effects of simvastatin in differentiating cells were not abolished by mevalonate or isoprenoid derivatives of mevalonate. Moreover, the sensitivity of differentiating cells to simvastatin regarding cell viability was about 7 times higher than that of proliferating cells. After induction of differentiation in the presence of 1 μM simvastatin for 2 days, IGF-1-induced activation of ERK1/2 and Akt was significantly decreased. Although mRNA expression of the IGF-1 receptor β-chain (IGF-1R β) did not change, protein level of the 200 kDa IGF-1Rβ precursor was significantly increased by simvastatin in a dose-dependent manner. Mevalonate did not abolish the effect of simvastatin on IGF-1Rβ expression. These results suggest that simvastatin decreases IGF-1 signaling via a regulation of the post-translational modification of IGF-1Rβ in an HMG-CoA reductase inhibition-independent manner.
The safety of an oil-degrading bacterium, C2 strain, was evaluated for utilization in an open system for bioremediation of oil-contaminated environments. The C2 strain was identified as Rhodococcus erythropolis by performing an alignment analysis of the whole 16S rRNA sequence. R. erythropolis was classified as a nonpathogenic (category 1) bacterium. Biological and biochemical properties of the C2 strain also confirmed its nonpathogenicity. The pathogenicity and basic ecotoxicity were studied in laboratory animals and in a variety of test species, respectively. General and inhalation toxicities were not detected; additionally, there was no evidence of skin irritation, mutagenic potential, eye irritation, skin sensitization, ecotoxicity or notable pathogenicity. The comparison of these results with human exposure levels and previously published data indicates that the C2 strain appears to be safe for utilization in bioremediation of polluted environments, requires no special occupational health precautions during the application process, and has a low environmental impact. This study suggests that the C2 strain could be suitable for bioremediation of oil-contaminated environments.
Nicardipine hydrochloride (Nic), a calcium channel antagonist, is used for the treatment of hypertension. In the present study, we estimated its effects on the levels and activities of hepatic cytochrome P450 isoforms in spontaneously hypertensive rats given p.o. with Nic at a dose of 0.5, 2.5, 5, or 12.5 mg/kg at 24-hr intervals for 14 days. Therapeutic effects on the development of hypertension were observed at doses of 5 and 12.5 mg/kg/day. Significant increases in the levels of mRNAs and enzyme activities of hepatic P450 isoforms, CYP1A1 and/or CYP1A2, by 14-day repetitive treatment with Nic were observed at lower therapeutic doses, whereas the increase in protein levels for CYP1A2 was observed at a higher therapeutic dose of 12.5 mg/kg/day. Likewise, the activities of hepatic CYP2B and CYP3A subfamily enzymes were increased by the 14-day-treatment of Nic only at a therapeutic dose (12.5 mg/kg/day), whereas their mRNA and protein levels were increased at lower therapeutic doses. To date, the dihydropyridine family, including Nic, has been believed to have inhibitory effects on the activity of various cytochrome P450 enzymes, especially human CYP3A4. However, the present findings demonstrate for the first time that Nic-repetitive treatments at a therapeutic dose result in significant increases in the expressions and activities of hepatic CYP1A, CYP2B, and CYP3A subfamily enzymes. Therefore, the effects of dihydropyridine family on cytochrome P450 enzymes have to be further validated to provide information on its safe and beneficial therapeutic application.
The anal fin in Japanese medaka, Oryzias latipes, is a typical sexual secondary character. In the present study, we focused on this organ and examined the effects of low doses of a natural estrogen, 17β-estradiol (E2), and an environmental xenoestrogen, bisphenol A (BPA), in vivo by monitoring estrogen receptor (ER) α gene expression. Groups of adult male and female medaka were immersed in 10 −9 M E2 or 10 −10 to 10−8 M BPA and the levels of ERα gene transcripts in the anal fins were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). One day of treatment with each concentration of BPA examined and 10 −9 M E2 increased the levels of ERα mRNA in female anal fins by 3-fold as compared with controls. In the male specimens, neither 10 −9 M E2 nor 10 −10 M BPA showed remarkable effects on the anal fins as compared with the results in females, but 10−9 and 10 −8 M BPA increased the levels of ERα mRNA by 2.3- and 3.3-fold with 1 day of exposure, respectively. The present results showed that medaka anal fins may be a sensitive bio-indicator for screening of environmental estrogenic chemicals.
Rhabdomyolysis has been reported after venlafaxine ingestion. We wished to characterize the prevalence of this adverse effect in a realistic clinical setting. Therefore, a retrospective casenote review was performed, including 235 patients admitted to the Royal Infirmary of Edinburgh due to venlafaxine overdose between January 2000 and June 2006. Seizures occurred in 8.9% of the study population. Patients who suffered seizures had ingested larger quantities of venlafaxine than those who did not develop seizures; median (interquartile range) 2800 mg (2006-4350 mg) versus 1500 mg (900-2700 mg, p = 0.001). Raised CK values were more prevalent in those with seizures than those without seizures (61.1% versus 25.7% respectively, p = 0.004). Nonetheless, a positive correlation was found between the quantity of venlafaxine ingested and CK across the whole group (rho = 0.201, 95% confidence interval 0.045-0.347), and in patients who had not developed seizures (rho = 0.174, 95% confidence interval 0.009-0.331). Venlafaxine overdose is associated with a high prevalence of acute muscle injury, both in patients who develop seizures and in those who do not. The clinical significance of this association merits further consideration.
To examine the transcriptional responses of rat kidney cells continuously exposed to cadmium, we performed DNA microarray analysis. Cadmium increased levels of expression of 27 genes, including genes for Mt1, Mt2, GSTa3 and B2m and reduced those of 4 genes.
To examine the transcriptional responses of rat bone cells continuously exposed to cadmium, we performed DNA microarray analysis. Cadmium increased levels of expression of 13 genes, including genes for Spp1, and reduced those of 10 genes.