The acute and chronic toxicity studies of aqueous extract (SPE) and seed powder (SPP) of Strychnos potatorum (S. potatorum) Linn were carried out in Wistar albino mice and rats, respectively. The animals did not show any toxic effects upto the dose of 2000 mg/kg, p.o. According to OECD guidelines - 423 for acute oral toxicity, the LD50 dose of 2000 mg/kg and above is categorized as ''unclassified.'' For chronic toxicity study, 100 and 200 mg/kg, p.o. of SPE and SPP were administered to Wistar rats for 90 days and various parameters like food and water intake, body weight changes, haematological parameters like red blood corpuscles (RBC), white blood corpuscles (WBC), haemoglobin (Hb) and erythrocyte sedimentation rate (ESR), biochemical parameters like blood glucose and urea, serum creatinine, enzyme parameters like alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and acid phosphatase (ACP) were studied. There were no significant changes in any of the above parameters of drug treated groups with respect to control group, which explain their nontoxic nature. Further the nontoxic effect of the drugs SPP and SPE were confirmed by histopathological examination of various organs like liver, kidney, spleen and heart. Phytochemical studies of the drugs showed the presence of carbohydrates, alkaloids, steroids/triterpenes, polyphenolics, saponins and polysaccharides in SPP and carbohydrates, steroids, triterpenes, saponins, polyphenolics and polysaccharides in SPE.
This study investigated the effects of mixtures of Vitis vinifera (V), Schizandra chinensis (S), Taraxacum officinale (T), Gardenia jasminoides (G), Angelica acutiloba (A), and Paeonia japonica (P) on fatty liver and hepatotoxicity induced by an ethanol liquid diet. Male Sprague-Dawley rats received a normal diet (ND), ethanol diet (ED), ED + VST (ethanol diet + V 100 + S 150 + T 150 mg/kg/day), ED + VGS (ethanol diet + V 100 + G 150 + S 150 mg/kg/day), ED + VGT (ethanol diet + V 100 + G 150 + T 150 mg/kg/day), and ED + VAP (ethanol diet + V 100 + A 150 + P 150 mg/kg/day). Rats fed liquid diets containing alcohol for 6 weeks showed remarkable increases in serum and hepatic lipid levels, indicating the onset of alcoholic fatty liver. Greater increases in alanine transaminase (ALT) and alkaline phosphatase (ALP) activities in serum were observed in the groups fed alcohol-containing diets compared with those in the ND group. Treatment with ED + VST, ED + VGS, ED + VGT, and ED + VAP decreased the levels of triglycerides, free fatty acids, and total cholesterol in the serum and liver, with a concomitant reduction in the activity of serum ALT and ALP. These data suggest that the plant extracts examined in this study can be utilized as a health functional food or new drug candidates for the treatment of fatty liver and hepatotoxicity induced by chronic alcohol consumption.
The mutagenic activity of size-fractionated atmospheric particulate matter (PM) was investigated at both a roadside site and a suburban site in Saitama, Japan. Sampling was carried out using a low-pressure cascade impactor between January and February 2005. The mutagenic activity of the size-fractionated PM was determined by the Ames test using Salmonella typhimurium strain TA98 without metabolic activation (S9). Most of the mutagenic activity was found in the fine particle fraction (< 2.1 μm) at both sites. There was almost no difference in the contribution of fine particles to the mutagenic activity of total PM between the two sites. On the other hand, there was a clear difference in the contribution of ultrafine particles (< 0.12 μm) to the mutagenic activity between the two sites. At the suburban site, ultrafine particles accounted for 5.7% of the mutagenic activity of total PM. In contrast, at the roadside site, ultrafine particles contributed as much as 12% to the mutagenic activity of total PM, although their contribution to the total PM mass was only 2.3%. Moreover, the mutagenic activity per unit air volume in the ultrafine particle fraction at the roadside site (1.2 revertants/m3) was 3.1-fold higher than that at the suburban site (0.38 revertants/m3), although the activity in the fine particle fraction at the roadside site was only 1.4-fold higher than that at the suburban site. These results indicated that, with regard to mutagenicity, the health risk due to ultrafine particles is relatively high at roadside areas.
Enterococcus faecalis (E. faecalis) ATCC51299 used as a control in screening for high-level aminoglycoside resistance in enterococci was observed to transfer plasmids to the clinically isolated E. faecalis strain 4437 in a broth medium. From a mixed culture of strain ATCC51299 and strain 4437, transconjugants were obtained not only from the planktonic phase but also from biofilm. Plasmid transfer frequency was approximately one order higher among cells in the biofilm than among those in the planktonic phase. When the biofilm formed by the recipient strain E. faecalis 4437 was inoculated with the donor strain E. faecalis ATCC51299, correlations were observed between the number of donor cells and transconjugants, and the number of donor cells and transfer frequency in the biofilm (r = 0.99, 0.98, respectively). In addition, the transfer frequency in biofilm remained constant after 9-hr incubation, while transfer frequency in the planktonic phase decreased with incubation time. These results suggest that the biofilm formed by enterococci on surfaces gives rise to efficient gene transfer.
The mouse counterpart of the human inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP), inter-alpha-trypsin inhibitor family heavy chain-related protein, was partially purified from mouse serum through Western blot analysis using anti-human IHRP antibody. The amino-terminal amino acid sequence was then determined, and the cDNA clone encoding mouse IHRP was isolated and characterized. The mouse IHRP amino-terminal amino acid sequence matched the predicted sequence from the cDNA, and has high homology compared with human and pig IHRP. Furthermore, the whole mouse IHRP amino acid sequence predicted from the cDNA nucleotide sequence showed reasonable homology to that of both human and pig IHRP. Mouse IHRP expression was induced by treatment with typical inflammation-inducing compounds, such as turpentine and lipopolysaccharide.
2-Alkylcyclobutanones in irradiated and cooked meats, poultry and egg were analyzed to detect the irradiation history of those samples prior to cooking. They were extracted using an accelerated solvent extraction system (ASE), and determined by gas chromatography with mass spectrometry (GC/MS). Irradiated beef, chicken and egg were browned and the irradiated beef and egg were boiled. The irradiated chicken was fried and a pancake was made with the irradiated egg. Radiolytic compounds of 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutane (2-TCB) were detected in all the cooked samples when the raw samples were irradiated with 3-6.5 kGy. Both 2-DCB and 2-TCB were similarly stable in the samples with conventional cooking, which resulted in a temperature less than 100°C inside the samples, whereas they were destroyed when heated to over 200°C. The food samples did not show serious interferences on GC/MS chromatograms due to cooking. The results conclude that the analysis of 2-DCB and 2-TCB in the cooked samples would be a reliable indicator to detect the irradiation history of raw meats, poultry and egg.
The immediate-type allergic reaction is involved in many allergic diseases such as asthma, allergic rhinitis, and sinusitis. The discovery of drugs for the treatment of immediate-type allergic disease is a very important subject in human health. The formulated ethanol extract of Artemisia asiatica Nakai (DA-9601) has been reported to have anti-oxidative and anti-inflammatory activities. In this report, we investigated the effect of DA-9601 on the immediate-type allergic reaction and studied its possible mechanisms of action, focusing on the mast cell-mediated allergic reaction. DA-9601 inhibited compound 48/80-induced systemic anaphylactic reactions and serum histamine release in mice. DA-9601 decreased the IgE-mediated passive cutaneous anaphylaxis, the model of local allergic reaction in vivo. DA-9601 dose-dependently reduced histamine release from mast cells activated by compound 48/80 or IgE. Furthermore, DA-9601 decreased the gene expression and production of tumor necrosis factor (TNF)-α in phorbol 12-myristate 13-acetate plus A23187-stimulated mast cells. These findings provide evidence that DA-9601 could be a candidate as an anti-allergic agent.
[Objective] To investigate the effect of cordyceps sinensis (CS) on expression of uncoupling protein-2 (UCP2) and so to elucidate the role of UCP2 in development of nonalcoholic fatty liver diseases (NAFLD). [Methods] Rats were administrated with high-fat diet to produce NAFLD animal model and intervened by cordyceps sinensis. Triglyceride (TG), total cholesterol (TC) in liver were measured by biochemistry, adenosine triphosphate (ATP) by luciferin-luciferinase, and UCP2 expression by Northern blotting and immunohistochemistry. Liver histopathology was evaluated. [Results] High-fat diet fed rats developed obesity and showed a gradual increase in body weight, liver index, and a decrease in ATP level. More advanced liver disease was found histopathologically for longer high-fat diet. Up-regulation of liver UCP2 by high-fat diet stopped after week 12. However codyceps sinensis induced UCP2 up-regulation continuously, and kept liver ATP a relatively high level. [Conclusion] NAFLD rat models were produced successfully. Liver UCP2 up-regulation in NAFLD rats may be a definite beneficial adaptation to lipid exposure. Cordyceps sinensis may serve a protective role to prevent NAFLD from progression. One of the possible mechanisms involves in modulating UCP2 expression and thereby, regulating fat metabolism, energy homeostasis.
Two plastic items were investigated for toxicity, due to chemical migrants, on reproduction and subsequent pregnancy outcomes. Extraction of high-density polyethylene (HDPE) food oil jerrycans, and polyvinyl chloride (PVC) blood bags was carried out. HDPE and PVC were extracted with sesame oil and normal saline, respectively. The extracts were prepared daily and administered (50 ml/kg b.w.) into pregnant Swiss albino mice from gestation day 0 until delivery. Control groups received the pure vehicles that were subjected to the same conditions of extraction and extracts. Pregnancy weight gain, gestation period, litter size, stillbirths and offspring sex ratio were recorded. Blood sex hormones (progesterone, estradiol and prolactin) were assayed for each pregnancy trimester. Birth weight, growth rate and sex hormone levels [females: follicle stimulating hormone (FSH), leutenizing hormone (LH) and estradiol (E2); males: testosterone] were monitored in offspring. ELISA was applied to assay hormones. HDPE caused significant (p ≤ 0.01) stillbirth. Blood hormone levels in dams and offspring for both treatments indicated no significance. PVC treatment exhibited negative effects on all parameters. In conclusion, HDPE is leachable and could affect reproduction, as indicated by the stillbirth incidence. PVC sample might not be toxic at the conditions of the experiment. Oil-plastic extract could exhibit a pronounced effect on pregnancy outcomes in contrast with the aqueous one.
We previously showed that several small-dose 0.5 Gy whole-body γ-ray irradiation inhibits tumor growth in mice via elevation of the interferon (IFN)-γ/interleukin 4 (IL-4) ratio concomitantly with a decrease in the percentage of B cells. Here, we examined whether repeated small-dose (0.5 Gy, 10 times) γ-ray irradiation influences atopic dermatitis in an NC/Nga mouse model. It was found that repeated γ-ray irradiation increased total IgE in comparison with the disease-control group. Levels of IL-4 and IL-5 were increased versus the disease-control group, while IFN-γ was slightly decreased, resulting in a further decrease of the IFN-γ/IL-4 ratio compared with the disease-control group. These results indicate that repeated small-dose γ-ray irradiation may exacerbate atopic dermatitis. This may be because the irradiation induces not helper T lymphocyte 1 (Th1), but Th2 polarization in this atopic mouse model, i.e., the effects of small-dose irradiation may be different in conditions involving immune hypersensitivity and impaired immunity.
The effects of Saldi tierra containing various trace elements on bone components in the femoral-diaphyseal and -metaphyseal tissues of rats was investigated. Saldi tierra (sodium salt or potassium salt) was obtained from natural zeolite using the method of ion exchange with sodium chloride or potassium chloride solutions. Saldi tierra contained more than 20 elements. Rats were orally administered a solution of Saldi tierra once daily for 7 days. Calcium content, alkaline phosphatase activity and DNA content in the femoral-diaphyseal (cortical bone) and -metaphyseal (trabecular bone) tissues of rats were significantly increased with the administration of Saldi tierra sodium salt (100 mg/100 g body weight). Calcium content and alkaline phosphatase activity in the femoral-diaphyseal tissues of rats were significantly increased with the administration of Saldi tierra sodium salt (50 mg/100 g). Saldi tierra (50 mg) contained 3.155 mg of calcium. Femoral components were not significantly altered by the administration of calcium chloride (Ca 3.155 mg/100 g body weight). Moreover, the administration of Saldi tierra potassium salt (50 or 100 mg/100 g) caused a significant increase in bone components in the femoral-diaphyseal or -metaphyseal tissues of rats. Potassium salt of Saldi tierra had a potent effect on bone components as compared with its sodium salt. Bone components were not significantly increased with the administration of an elemental mixture containing sodium (26.8 mg), potassium (1.660 mg), magnesium (0.431 mg), chloride (43.86 mg), silicon (11.1 μg), calcium (6.310 mg), zinc (0.42 μg), and manganese (1.32 mg/100 g), which are contained in Saldi tierra (100 mg). This study demonstrates that the oral administration of Saldi tierra containing various elements has anabolic effects on bone component in rats. The intake of Saldi tierra may have a role in the prevention of bone loss with aging.
Prostate cancer has become the most common male cancer in Western countries and the incidence of this disease is increasing steadily in developing countries including China. The aim of the present study was to determine whether milk consumption promotes the development of prostate carcinogenesis in rats since milk consumption is considered to be a risk factor in some epidemiological studies. In the present study, we compared the prostate carcinogenesis in rats fed on commercial whole milk and artificial milk with or without induction by amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). The atrophic changes became more serious in the ventral prostate in the milk groups with or without PhIP administration than in the corresponding artificial milk groups. In the ventral prostate, the incidence of prostatic intraepithelial neoplasia (PIN) also was significantly higher in the milk group than in the artificial milk group irrespective of PhIP administration. The rats given PhIP, either fed on milk or artificial milk, showed significantly higher incidences of PIN and adenocarcinoma in the ventral prostate, and dysplasia in the seminal vesicle. The present study suggests that commercial whole milk is likely to promote the development of prostate, especially ventral prostate, carcinogenesis in rats.
The in vivo metabolism of 5-methoxy-N,N-diisopropyltryptamine (5-MeO-DIPT), a psychoactive tryptamine analog, was studied in rat. Male Wistar rats were administered 10 mg/kg 5-MeO-DIPT hydrochloride orally, and urinary fractions were collected. After enzymatic hydrolysis, the metabolites were extracted by liquid-liquid extraction and analyzed by gas chromatography/mass spectrometry. 5-Methoxy-N-isopropyltryptamine, 5-hydroxy-N,N-diisopropyltryptamine (5-OH-DIPT), 5-hydroxy-N-isopropyltryptamine, and 5-methoxyindole-3-acetic acid were identified as 5-MeO-DIPT metabolites. By quantitative analysis using high-performance liquid chromatography, it was revealed that 5-OH-DIPT was the main metabolite of 5-MeO-DIPT in rat, comprising 20.5% of the dose administered. On the other hand, only 0.8% of 5-MeO-DIPT administered was excreted into the urine in its original form.
This paper focuses on the stochastic analysis of the daily variations in formulations dispensed at pharmacies. A recent study showed that the cross-correlation functions between the time variations can be used for estimating the route and speed of influenza propagation process in society. In reality, however, the peak-shaped signals of the time variations and cross-correlation function are so complicated and noisy that it is almost impossible to know the original shape of the signals. The aim of this paper is to give an answer to this problem by considering the mathematical relationship between the time variations and cross-correlation function. Influenza anti-viral agents for adults and children are taken as an example.
The cross-correlation functions between the time series of drug sales at community pharmacies have been used for the epidemiological understanding of influenza propagation process. The aim of this paper is to examine the methods of smoothing for determining the maximum position (time lag) of the cross-correlation function. The moving average method with a window of seven days is taken to eliminate the hebdomadal frequencies originating from the life style of people around the pharmacies, or rather, purchasers. Based on the smoothed data of influenza anti-viral agents for adults and children at pharmacies in Tokyo and its vicinity, this paper tries to give an answer to the question of which are earlier infected with influenza, adults or children, in the limited areas.
In Japan, five new drug applications (NDAs) of triptans have been approved through a bridging strategy. They constituted the largest target disease field among 26 NDAs which had been approved through the strategy between 1999 and 2003. The bridging strategies of the drugs were classified into two major categories to be described below. One was to conduct a placebo-controlled dose-response study as a bridging study in an attempt to extrapolate the data from the pivotal foreign Phase III studies including a repeated dose study (i.e., zolmitriptan, sumatriptan succinate, and eletriptan hydrobromide). Another was to conduct a placebo-controlled Phase III study in Japan in an attempt to extrapolate the data for efficacy from a repeated dose study (i.e., sumatriptan and rizatriptan benzoate). The extrinsic ethnic factors relating to triptans did not interfere with the extrapolation of foreign clinical data in the five applications. A bridging strategy reduced the number of clinical trials and/or sample size. The accumulation of these bridging experiences indicated that foreign clinical data could be used to support the approval of triptans in Japan.
We investigated time-dependent changes in the fate of methylmercury (MeHg) in both sexes of hairy and hairless mice during 6 days after its single administration to clarify whether the presence of hair could affect tissue distribution and excretion of MeHg. Despite the excretion of mercury (Hg) into hair of hairy mice, Hg concentrations in several but not all tissues were higher in hairy than in hairless mice, especially in the brain, liver, kidney and testes in males and in the brain, liver and blood in females. The cumulative amounts of Hg excreted in feces in males and in both urine and feces in females were lower in hairy than in hairless mice, whereas there was no significant difference in the urinary level in males. However, significant differences in the tissue Hg levels were observed earlier than those in the excretory Hg levels. Accordingly, the higher Hg levels in various tissues of hairy mice as compared to those in hairless mice would not have resulted from the lower Hg excretion levels. These results suggest that the presence or absence of hair markedly affects the fate of MeHg, but the differences in its fate between hairy and hairless mice would not be directly due to the fact that hair provides an excretion route.
Rice bran was found to adsorb Fe3+ effectively over the range of pH 1-12. Therefore rice bran is applicable for uses in the food industry over a wide pH range. Fe3+ was successfully removed from water samples with an average removal efficiency of 88.5% after 90 min when rice bran was added to water samples containing 5 mg/l Fe3+. The removal of Fe3+ by rice bran was attributed to the chelating properties of phytic acid. Here, we report a fundamental study on the efficiency of rice bran for the removal of Fe3+ using a batch system on a laboratory scale and describe the elucidation of the mechanism of Fe3+ removal by rice bran.
A wound dressing material, SG-01 was developed to treat various skin ulcers. In this study, to clarify the features of SG-01, we investigated the therapeutic effects of this dressing material in rat burn and decubitus ulcer models. In the rat burn model, SG-01 significantly reduced the percent wound area compared to gauze (control group), and decreased the sum of the percent area, suggesting that SG-01 promotes the healing of burns. Subsequently, we investigated the curative effects of SG-01 in the rat decubitus ulcer model using commercially available dressing materials, NU-GEL®1,2) and DuoACTIVE CGF®.3,4) SG-01 significantly reduced the percent wound area compared to gauze (control group), decreased the sum of the percent area, and shortened the interval until healing. There were no significant differences between SG-01 and the above commercially available materials. These results suggest that SG-01 is useful for treating burn and decubitus ulcers.
Pollution in the Watarase River caused by mineral wastewater containing high levels of copper discharged during the development of the Ashio Copper Mine is one of the most well-known environmental pollution problems in Japan, and has been called the ''Starting line of environmental pollution problems in Japan.'' In this study, we conducted follow-up investigations on the conditions of pollution in the Watarase River since 1991, and measured the heavy-metal levels in water in other rivers near the Watarase River and in soils around them. In addition, we compared the results in this study with previous results, i.e. pollution conditions in the Watarase River, other rivers and soils. Six points in upstream sites of the Watarase were chosen for sampling. Six samples from river water, five samples from soils around the respective points, and three samples from tap water distributed there were collected for the analysis. Heavy metal levels in rivers and soils around the Ashio Copper Mine were significantly lower than the environmental standards for them. When compared with our previous investigations, the levels of polluting heavy metals around the closed refinery at the Ashio Mine were gradually but clearly reduced. In conclusion, the environment around the Watarase River has been steadily improved so that safety in the living environment is assured.
Species differences in the induction of hepatic cytochrome P4501A (CYP1A) subfamily enzymes by nitroanisidines, such as 2-methoxy-4-nitroaniline (2-MeO-4-NA), 2-methoxy-5-nitroaniline (2-MeO-5-NA), and 4-methoxy-2-nitroaniline (4-MeO-2-NA), were investigated among male F344 rats, C57BL/6 Cr mice, and Hartley guinea pigs. All species of animals were treated with a single intraperitoneal injection of each chemical (0.44 mmol/kg body weight), and changes in hepatic microsomal activities for methoxyresorufin O-demethylation (MROD) and ethoxyresorufin O-deethylation (EROD), which were mainly catalyzed by CYP1A2 and CYP1A1, respectively, were examined. Gene expression levels of hepatic CYP1A subfamily enzymes were also examined by a reverse transcription-polymerase chain reaction (RT-PCR) method. These results indicated that all the chemicals examined showed abilities to induce hepatic CYP1A subfamily enzymes in rats but not in the other examined animal species. In the rat liver, the abilities of the chemicals to induce microsomal MROD activity and CYP1A2 gene expression were in the order 2-MeO-4-NA > 2-MeO-5-NA > 4-MeO-2-NA, whereas their abilities to induce microsomal EROD activity and CYP1A1 gene expression were in the order 4-MeO-2-NA > 2-MeO-5-NA > 2-MeO-4-NA. These findings demonstrate for the first time that there are species differences in the induction of hepatic CYP1A subfamily enzymes by the nitroanisidine isomers among rats and other rodents, mice, and guinea pigs.
To investigate the role of aromatic amino acids of MerT protein in phenylmercury transport, two merT variants (pMRTm1P and pMRTm2P) with specific site-directed mutations were constructed and examined their effects on C6H5Hg+-transport. Substitution of Phe-36 and Trp-40 residues located on the periplasmic loop of MerT in turn with Ala and Val, respectively, did not affect the Hg2+-uptake, but caused a significant reduction in the C6H5Hg+-uptake by bacterial cells with intact merT gene. Introduction of specific mutations changing Phe-108, 114, 115 to Ala, and Tyr-110, 116 to Ser in the C-terminal region of the third transmembrane of MerT also caused large reduction in the uptake of C6H5Hg+, but had no effect on the Hg2+-uptake. In addition, both mutations caused a significant reduction in the hypersensitivity to C6H5Hg+, but without affecting the Hg2+-hypersensitive phenotype. Together these results suggest that the aromatic amino acids of MerT protein may play an important role in the transport of C6H5Hg+ across the cell membrane.
Aim: Approximately 10-20% of general population has irritable bowel syndrome (IBS) and/or urolithiasis (U) and it seems that there is a significant association between them. Methods: We randomly took consecutive patients with and without IBS, among the cases applying to internal medicine polyclinic. U was diagnosed either by medical history or current findings. Results: Female/male ratio of IBS cases was 1.62. On the other hand, U was detected in 111 cases, totally, and female/male ratio was 0.94. So the rates of U were 13.7 and 15.2% in female and males, respectively. As a significant finding, U was detected in 61 (17.9%) of cases with IBS, whereas this ratio was 11.6% (50 in number) among the cases without (p < 0.01). On the other hand, there was no significant difference between groups of IBS and U according to obesity and hyperuricemia (p > 0.05 for both). Conclusions: Although IBS predominantly keeps females, relationship between IBS and U is still significant. IBS is probably a cascade of many physiological events, being initiated with infection, inflammation, psychological disturbances like many stresses and eventually terminating with gut dysfunction. So U may be one of the terminating points of the physiological events' cascade, IBS. By this way, the giant gap about the underlying etiologies of most of U cases may be explained by the high incidance of IBS in society. Keeping in mind this association will be helpful during prevention, treatment, and follow up of these pathologies.
A liquid coffee drink containing mannooligosaccharides from coffee mannan (MOS) was administered to a group of healthy adults. Subsequently, the amount of fat excreted from the body was examined. The subjects were divided into two groups: One was administered a liquid coffee drink containing MOS 3.0 g/day whereas the other was administered a placebo drink for seven days. Both groups were fed the standardized meals during the experiment. In the amount of average excreted fat, the drink containing MOS intake showed a significant increase in comparison with the before intake group and placebo intake group (p < 0.05 and p < 0.001, respectively). In addition, fat utilization in MOS intake group was significantly lower than the before intake group and the placebo group (p < 0.05 and p < 0.001, respectively). These results suggested that the intake of MOS 3.0 g/day increased in the amount of excreted fat and decreased in fat utilization.
We have already demonstrated many diesel exhaust particles (DEPs) and some damages in brain tissues (cerebral cortex and hippocampus) of newborn mice whose mothers inhaled DE during pregnancy, and these damages have the possibilities to lead to some disorders of nervous system. In this study, we examined pathological effects on newborn brain by DE-exposure to pregnant mice, especially focused on autism. ICR pregnant mice were exposed to DE and some were exposed to clean air as a comparative control. Brain tissues (cerebellum) were obtained from the mice (housed in a clean air) born from DE-exposure and control pregnant mice, and examined with light and electron microscope. To detect apoptosis, the immunohistochemical staining for caspase-3 was performed, especially; the numbers of positive Purkinje cell in cerebellum were compared between DE-exposure and control. In DE-exposure group, numerous caspase-3 positive cells were diffusely observed and the number of positive Purkinje cells was significant large than in control. Electron microscopically, specific features of apoptosis were found in Purkinje cells in the DE-exposure group. These findings indicate that DE-exposure to pregnant mice has a severe impact on fetal brain development and, especially, numerous apoptotic Purkinje cells cause the innate deficiency of them and would involve the pathogenic backing of autism. Our results would give a grave warning that the maternal inhalation of DE is hazardous to fetuses' health and it is possible that these fetal damages carries a great risk of various disorders of nervous system afterward, such as autism.
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