Effect of toluene inhalation on astrocytes and neurotrophic factor in rat brain

Toluene, an abused substance in Japan, is a neurotoxic chemical that has been shown to have neurobehavioral and electrophysiological effects. In previous work, both acute and chronic effects of toluene on cells have been studied extensively. However, although glial cells are thought to play an important role in the survival of neurons in the brain, the effect of toluene on glial cell function has not yet been characterized. To elucidate this, the effect of toluene inhalation on astrocytes in rat brain was examined. Toluene exposure (1500 ppm for 4 h on 4-10 days) augmented glial fibrillary acidic protein (GFAP) immunoreactivity, particularly in the hippocampus and cerebellum. Quantitative analysis showed that toluene inhalation markedly enhanced GFAP expression in the hippocampus and cerebellum. In both regions, proliferating cell nuclear antigen (PCNA) showed no obvious changes, but glutamine synthetase (GS)-immunoreactive cells were markedly increased by toluene exposure. Thus, the elevation of GFAP expression was induced by astrocyte activation rather than by cell proliferation. If toluene exposure activates astrocytes, astrocytes may play a role in the neurophysiological changes observed in toluene intoxication. A neurotrophic factor, basic fibroblast growth factor (b-FGF) was observed immunohistochemically in the capillary vessel walls in the hippocampus and the cerebellum of toluene-intoxicated rats. Basic-FGF may have induced GFAP expression both in the hippocampus and the cerebellum. So, other neurotrophic factors may affect the difference of GFAP elevation between the hippocampus and the cerebellum. These differences may relate to neurobehavioral function of each brain part after toluene exposure.     

Anger feelings and anger expression as a mediator of the effects of witnessing family violence on anxiety and depression in Japanese adolescents

The effects of anger feelings (rated by the State-Trait Anger Expression Inventory) and witnessing family violence on anxiety and depression (rated by the Hospital Anxiety and Depression Scale) were examined in 457 junior high school students. Anxiety and depression scores were correlated with frequencies of witnessing family violence. In a regression analysis, however, after controlling for the demographic variables and depression score, the anxiety score was predicted by State Anger, Anger-Out, and Anger-Control; the depression score was predicted, after controlling for the demographic and anxiety score, by State Anger, Anger-In positively, and by Anger-Out and Anger-Control negatively. Witnessing family violence failed to add significant contribution in predicting anxiety or depression. These results were generally supported by structural equation modeling. The effects of witnessing family violence on dysphoric mood may be mediated by anger feelings. The style of dysphoric mood may be predicted by the style of anger expression.     

Institutional Reactions to the Flag State that has Failed to Discharge Flag State Responsibilities

Netherlands International Law Review - The international regulatory system for the oceans is to a considerable extent dependent upon the effective exercise of flag state responsibilities but...     

Authors Index Volume 10

Authors Index

Authors Index Volume 10     

Fast PCR amplification using AmpFlSTR Identifiler: Second report

At ISFG2007, an earlier conference, we presented reports on two fast PCR cycling methods using AmpFlSTR Identifiler. In our current study, which involved PCR amplification using AmpliTaq Gold Fast PCR Master Mix, UP, and a standard PCR thermal cycler rather than the Fast PCR thermal cycler, we succeeded in allele typing while reducing PCR running times by half (to approximately 90 min).     

Case study of dissolving allele dropout

Allele typing performed using the original components of the AmpFlSTR Identifiler kit led to unbalanced peak heights and allele dropouts. However, allele typing was completely successful when substituting the QIAGEN Multiplex PCR Master Mix for the AmpFlSTR PCR Reaction Mix of the Identifiler kit. The successful range for allele typing was 250 pg to 1.5 ng of DNA. The running times for PCR amplification were reduced by about 30 min from the original protocol.     

Detection of diatom in formalin-fixed tissue by proteinase K digestion

The diatoms detection has been proposed to be useful in the diagnosis of drowning. Enzymatic digestion of unfixed lung tissues and other organs with proteinase K is widely employed to detect diatoms. Handling unfixed organs or blood from the bodies with some infectious diseases could prove to be dangerous. In this study, we examined the application of enzymatic digestion for diatom detection to formalin-fixed lung obtained at autopsy. Furthermore, we assessed the effect of hydrogen peroxide on the contamination of the lung specimen with foreign bodies inhaled in the course of drowning, smoking, or air pollution. Formalin-fixed lung was heated in 0.01 M Tris–HCl buffer (pH 7.5) containing sodium dodecyl sulfate (SDS) (tissue lysis-buffer), with or without glycine. Thereafter, the lung was subjected to enzymatic digestion with proteinase K. A part of formalin-fixed or unfixed samples digested with proteinase K were incubated with hydrogen peroxide at 80 °C for 6 h or 12 h, while the residues were processed without incubation. Formalin-fixed samples heated in tissue lysis-buffer with glycine could be digested with proteinase K; further, the number and proportion of diatoms detected in both formalin-fixed and unfixed samples were observed to be similar. The results suggest that enzymatic detection of diatoms can be applied to formalin-fixed organs by heating the samples in glycine-containing tissue lysis-buffer. As the use of formalin-fixed tissue for diatom detection can decrease risk of contamination by pathogenic organisms during the course of enzymatic digestion, the method presented in this study would be beneficial, to some extent, to individuals performing diatom analysis. Moreover, our results suggest that archival organs stored in formalin solution could be available in diatom detection over a long time-period following autopsies. Clearer image of diatoms was observed in the specimen incubated with hydrogen peroxide for 6 h, in which inhaled foreign bodies were discolored, than those not subjected to incubation. Therefore, incubation of sample digested with hydrogen peroxide in the limited time would be helpful for quantitative and qualitative diatom analysis.     

Providing credibility around the world: effective devices of the Cape Town Convention

This paper analyzes the characteristics and effects of the Cape Town Convention, which provides legal protection for investors in relation to asset-based aircraft financing. Some countries lack credibility for investors, who believe that the protection level of creditor rights in the country differs before and after investment. This time inconsistency problem results in credit rationing. Airlines in such countries cannot access the credit market, particularly the international capital market. This paper first examines why developing countries have been suffering from a lack of credibility and from credit rationing, and then explores how the Convention resolves this problem. We focus on two devices: the white list system and the system of declarations. The former gives the contracting states incentive not to break the Convention. The latter is considered to promote the ratification of the Convention by potential contracting states. These devices enable the Convention to attract a number of states.