Search

Bilinguals are often determined by how early they are exposed to L2 and the length of their immersion in the L2 environment, both of which affect factors that are important for acquiring language. To analyze the relationship between these, the language function of English-Japanese bilinguals was assessed using fMRI. In order to aim to observe differences in the brain, language function areas were activated by tasks specific to L2 usage. There were 24 English-Japanese bilingual speakers, average age of 25, categorized into four groups according to the age of exposure (before or after age 8), and the length of exposure (longer or shorter than 5years):Early Long (EL), Early Short (ES), Late Long (LL) and Late Short (LS). A PHILIPS Achieva 3.0T TX MRI machine was used to scan the subjects. Tasks consisted of hearing a short story in Japanese (Issunboshi) and English (Peter Rabbit) and their reverse plays by comparison of BOLD oxygenation levels. Focus was given to Broca’s area, Wernicke’s area and the premoto...

Holmes’ tremor is a rare symptom characterized by a complex combination of resting, postural and intention tremor. A 66-year-old right-handed man post-traumatically developed severe action tremor in his right hand, which had a component of the re-emergent type and predominantly involved proximal muscles. He also presented mild resting tremor in his right hand. As the response of levodopa/carbidopa and arotinolol was limited, he started zonisamide, which resulted in dramatic improvement. The Fahn–Tolosa–Marin Tremor Rating Scale motor score improved from 52 points to 24 points. In conclusion, zonisamide could be a therapeutic option for post-traumatic Holmes’ tremor. Further prospective studies are required.

The object of this study is to investigate the relationship between a typical product of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8OHdG), and mutagenesis in V79 cells through a molecular analysis of hypoxanthine-guanine phosphoribosyltransferase (hprt) gene mutants. We performed a direct sequencing analysis of the cDNA of mutants obtained after treatment with N,N'-bis(2-hydroxyperoxy-2-methoxyethyl)-1,4,5,8-naphthalenetetracarboxylic-diimide (NP-III) or riboflavin, each of which induces the formation of 8OHdG in cellular DNA upon UVA irradiation. The frequency of mutation after both treatments was no more than 2 to 5 times the control value. A considerable number of the mutants could not be amplified by RT-PCR, and this was also the case for the control mutants. Among the mutants analyzed, deletions and a TA-->AT transversion occurred predominantly. The reasons for the weak association of induction of 8OHdG with frequency of mutation and the possible mechanism of oxidative-stress-derived mutagenesis are discussed.

Immunohistochemical artifacts for nitrotyrosine were investigated in eosinophils with regard to fixatives. Immunoreactivity for nitrotyrosine was revealed in separated eosinophils and in gastric mucosa fixed with periodate, lysine-paraformaldehyde (PLP). The increase in immunoreactivity by PLP was due to periodate itself, a component of PLP. Nitrotyrosine formed by peroxidase using NO2- and H2O2 or by peroxynitrite was not completely inhibited by 100 mM dithionite but the immunoreactivity for nitrotyrosine antibodies by PLP was completely inhibited by 5.7 mM dithionite. Although untreated eosinophils or ovalbumin (OVA) did not show protein tyrosine nitration in a standard Western blot, the treatment of the blotted membrane with PLP increased the reactivities of proteins from eosinophils with anti-nitrotyrosine antibodies. The increase in immunoreactivity of OVA with anti-nitrotyrosine antibodies by PLP did not change with pre-treatment with dithionite but was abolished by treatment with dithionite after PLP fixation. In HPLC assays, periodate did not generate nitrotyrosine from L-tyrosine and aminotyrosine. These results suggest that the treatment of eosinophils or eosinophil-containing tissues with PLP fixative augments the immunoreactivity of nitrotyrosine antibodies with eosinophils due to the formation of epitopes similar to nitrotyrosine by an oxidation reaction of periodate, which evokes an artifact in nitrotyrosine immunohistochemistry.

To examine changes in the Body Mass Index (BMI) and in the prevalence of obesity among male Japanese workers after five years.Follow-up study, based on data from annual health checkups performed at workplaces.We had access to the data for a total of 63,951 male workers aged 20 through 54 years in 1992 who were examined in both 1992 and 1997. In quinquennial cohorts, we compared the findings for mean BMI and the prevalence of obesity in 1992 with the findings in 1997.Between 1992 and 1997, in all cohorts the mean BMI increased significantly. For each adjacent cohort pair the mean BMI value in 1997 was significantly higher than in 1992. In 1992, 21.3% of all subjects were obese, increasing by 3.4% after five years to 24.7% in 1997. In the younger 1968-1972 and 1963-1967 cohorts the prevalence of obesity increased by 6.3% during five years. The significant increase in the obesity prevalence was observed from 1992 to 1997 for each age-adjusted cohort.These results suggest a substantial increase in obesity among male Japanese workers. The notable increase in the prevalence of obesity among young male adults and the general trend toward widespread obesity during five years signal an increasing risk of lifestyle-related diseases.

Objectives Pain is one of the common symptoms in patients with Parkinson's disease (PD), with a prevalence of approximately 40–85%. These symptoms affect the quality of life of PD patients. We evaluated the effect of spinal cord stimulation (SCS) to chronic pain and motor symptoms of PD. Materials and Methods Three PD patients were treated with SCS to relieve their persistent and intractable pain. One patient had failed back surgery syndrome and the other two had lumbar canal stenosis. All patients had a stooped posture and pain that was resistant to analgesics. We evaluated motor symptoms using Hoehn and Yahr scale and the Unified Parkinson's Disease Rating Scale (UPDRS), and evaluated pain using visual analog scale and widespread pain index, before and after SCS. Results After SCS insertion, chronic pain in the patients decreased in both the lower back and limbs. Moreover, SCS ameliorated the symptoms of PD. One-year follow-up after SCS showed that UPDRS part III scores, rigidity, and tremor were improved without large alterations in levodopa dosage. Dementia and activities of daily living did not improve after SCS. Discussion and Conclusion Our results indicate that SCS may be a treatment option for both motor symptoms and chronic pain in PD, especially in cases complicated with lumbar canal stenosis or disc herniation. Further studies are needed to evaluate the efficacy of SCS in PD patients.

Purpose Previous studies suggest that compression and stretching of the corticospinal tract (CST) potentially cause treatable gait disturbance in patients with idiopathic normal pressure hydrocephalus (iNPH). Measurement of axon diameter with diffusion MRI has recently been used to investigate microstructural alterations in neurological diseases. In this study, we investigated alterations in the axon diameter and intra-axonal fraction of the CST in iNPH by q-space imaging (QSI) analysis. Methods Nineteen patients with iNPH and 10 age-matched controls were recruited. QSI data were obtained with a 3-T system by using a single-shot echo planar imaging sequence with the diffusion gradient applied parallel to the antero-posterior axis. By using a two-component low-q fit model, the root mean square displacements of intra-axonal space ( = axon diameter) and intra-axonal volume fraction of the CST were calculated at the levels of the internal capsule and body of the lateral ventricle, respectively. Results Wilcoxon's rank-sum test revealed a significant increase in CST intra-axonal volume fraction at the paraventricular level in patients (p

Natural Killer Cell Activity and its Related Psychological Factor, Sense of Coherence in Male Smokers: Hiroyuki NAKAMURA, et al. Department of Environmental and Preventive Medicine, Graduate School of Medical Science, Kanazawa University—Sense of Coherence (SOC), a candidate for a mediating stress factor, is studied in the framework of Antonovsky''s salutogenesis paradigm, which emphasizes the origin of health. Emotional stress is associated with a decrease in cellular immune function including natural killer cell activity (NKCA). The purpose of this study was to clarify the involvement of SOC and smoking behavior in cellular immunity including NKCA. The author performed a psychological questionnaire and examined NKCA and NK cell subsets in 125 men (45.1 ± 7.88 yr, mean ± SD) engaged in office work. The psychosocial factors included SOC and health locus of control (HLC) as well as perceived life stress and life style. SOC and smoking status was recognized to affect NKCA as well as CD57+CD16+. A higher SOC and never smoking significantly contributed to a higher level of NKCA. In subjects with a higher SOC, NKCA in never and ex-smokers were significantly higher than those in present smokers, whereas in subjects with a lower SOC, NKCA only in never smokers was significantly higher than that in present smokers. There were no significant differences in the stress index, alcohol obesity, or HLC among smoking status. SOC showed a moderately negative relationship with life stress. Multiple regression analysis demonstrated that SOC and smoking were significantly correlated to NKCA and CD57+CD16+, independently of other psychosocial variables, suggesting salutogenic contributions of SOC. SOC may be an important psychological modifier in determining the relationship between cellular immunity and smoking cessation. The reduced NKCA with lower SOC in present smokers, whose association seems to be produced not through daily life stress, must be dealt with as an occupational mental health problem. (J Occup Health 2001; 43: 191-198)

A numerical analysis of two-and three-dimensional separated and reattached flow and heat transfer in an enlarged channel is presented in this paper. Numerical calculations of Navier-Stokes equations and energy one are carried out using the finite difference method. The results of three-dimensional calculations are compared with the two-dimensional ones and show the effects of aspect ratio. The transition from symmetric to asymmetric flow is caused at lower Reynolds number with increasing the aspect ratio. The details of local heat transfer characteristics accompanying two different separated flow regions on two downstream walls are clarified. Two-dimensionality of the flow and heat transfer is disappeared at the aspect ratio considered.

The present paper shows a numerical analysis of two-dimensional unsteady separated and reattached flow and heat transfer around a rectangular cylinder located on a wall in a channel with two parallel walls. The Reynolds number ranges from 100 to 1000. For Rel300, the flow is found to be steady. For Re=400∼1000, the separated shear layer becomes unstable accompanying the formation of spanwise vortices, and in a range of Reg500, the reattachment length steeply decreases as Re increases. These vortices grow behind a cylinder and are shed almost periodically. The details of the local heat transfer characteristics along with the flow behaviors are clarified.

A numerical analysis based on the finite difference method was made on a three-dimensional separated and reattached flow around a surface-mounted hexahedron in a channel with two parallel walls, and the heat transfer characteristics therein were also analyzed. Reynolds numbers treated are 100∼1000 and Prandtl number 0.7, respectively. It is clarified from the numerical results that the flow around the hexahedron is very complicated, forming a horseshoe vortex around it, and also producing the separation and reattachment of flow on and downstream of it. The heat transfer characteristics over the hexahedron greatly depend on the flow behavior. These flow and heat transfer features vary with the Reynolds number.

A girl aged 2 years 10 months suddenly went into a deep coma and demonstrated left hemiplegia. At birth, she had exhibited a left-sided facial port-wine stain typical of Sturge-Weber syndrome (SWS) and involving the V1 and V2 distributions of the trigeminal nerve. Computed tomography showed a right thalamic hemorrhage with acute hydrocephalus. Magnetic resonance imaging with Gd enhancement 8 months before the hemorrhage had shown a patent superior sagittal sinus (SSS) and deep venous system. Magnetic resonance imaging and MR angiography studies 2 months before the hemorrhage had revealed obstruction of the SSS and right internal cerebral vein (ICV). Given that a digital subtraction angiography study obtained after the hemorrhage did not show the SSS or right ICV, the authors assumed that impaired drainage was present in the deep venous system at that stage. The authors speculated that the patient's venous drainage pattern underwent compensatory changes because of the occluded SSS and deep venous collectors, shifting outflow through other cortical venous channels to nonoccluded dural sinuses. Sudden congestion (nearly total to total obstruction) of the ICV may have caused the thalamic hemorrhage in this case, which is the first reported instance of pediatric SWS with intracerebral hemorrhage and no other vascular lesion. Findings suggested that the appearance of major venous sinus occlusion in a child with SWS could be a warning sign of hemorrhage.

We developed a high-throughput Enzyme-linked immunosorbent assay (ELISA) for measuring α2,6-sialylated transferrin (Tf), based on inhibition of anti-Tf antibody binding to α2,6-sialylated Tf by a lectin, Sambucus sieboldiana Agglutinin (SSA). The inhibition was not observed with other glycoforms, such as periodate-treated, sialidase-treated and sialidase/galactosidase-treated Tf, suggesting that the assay was glycoform specific. This finding was applied to an automated latex-agglutination immunoassay, using SSA lectin as an inhibitor (SSA-ALI). The concentration of α2,6-sialylated Tf measured by SSA-ALI in human cerebrospinal fluid was correlated with that of ELISA (r2 = 0.8554), previously developed for measuring α2,6-sialylated Tf.

Leucine-rich α2-glycoprotein (LRG) is a protein induced by inflammation. It contains a leucine-rich repeat (LRR) structure and easily binds with other molecules. However, the function of LRG in the brain during aging and neurodegenerative diseases has not been investigated. Here, we measured human LRG (hLRG) concentration in the cerebrospinal fluid (CSF) and observed hLRG expression in post-mortem human cerebral cortex. We then generated transgenic (Tg) mice that over-expressed mouse LRG (mLRG) in the brain to examine the effects of mLRG accumulation. Finally, we examined protein-protein interactions using a protein microarray method to screen proteins with a high affinity for hLRG. The CSF concentration of hLRG increases with age and is significantly higher in patients with Parkinson's disease with dementia (PDD) and progressive supranuclear palsy (PSP) than in healthy elderly people, idiopathic normal pressure hydrocephalus (iNPH) patients, and individuals with Alzheimer's disease (AD). Tg mice exhibited neuronal degeneration and neuronal decline. Accumulation of LRG in the brains of PDD and PSP patients is not a primary etiological factor, but it is thought to be one of the causes of neurodegeneration. It is anticipated that hLRG CSF levels will be a useful biomarker for the early diagnosis of PDD and PSP.

To clarify the mechanisms of intracellular induction of oxidative DNA damage, we have investigated the concentrations of intracellular reactive oxygen species and the amounts of 8-hydroxydeoxyguanosine (8OHdG), a mutagenic oxidative DNA damage, in human neutrophil-like cells, dimethylsulfoxide-differentiated HL60 (DMSO-HL60). We determined intracellular concentrations of hydrogen peroxide and superoxide by flow cytometry with dichlorofluorescein diacetate and hydroethidine, respectively. We determined the 8OHdG amounts with an electrochemical detector connected to HPLC after anaerobic sample processing. DMSO-HL60 releases superoxide upon stimulation with phorbol myristate acetate, and the released superoxide dismutates to hydrogen peroxide. Stimulation of DMSO-HL60 with 100 nM phorbol myristate acetate increased intracellular hydrogen peroxide, superoxide and 8OHdG (control). Addition of 1000 U/ml catalase decreased hydrogen peroxide (31.3% of control) and 8OHdG (20.3%). Addition of 100 U/ml SOD decreased superoxide (18.7%) and 8OHdG (41.6%). Addition of 1 mM deferoxamine decreased 8OHdG (30.4%), but increased hydrogen peroxide (129.6%). Addition of 200 microM 4-acetamido-4'- isothiocyanostilbene-2,2'-disulfonic acid decreased superoxide (59.9%) and 8OHdG (42.0%). Addition of 0.4% ethanol had no effect on superoxide concentration (102.2%), but tended to decrease hydrogen peroxide (83.5%) and 8OHdG (84.3%). Pretreatment of DMSO-HL60 with 0.1 mM FeSO4 increased 8OHdG (117.3%), but decreased hydrogen peroxide (75.8%). These findings indicate that the extracellularly released superoxide and hydrogen peroxide diffuse into the cell, but that such reactive oxygen species are not the direct molecules to induce 8OHdG. Our results suggest that 8OHdG is induced by the hydroxyl radical which is generated from intracellular hydrogen peroxide and superoxide-reduced Fe.

To establish an accurate 8-hydroxydeoxyguanosine (8OHdG) determination system, we examined two potential factors causing experimental error in 8OHdG determination. First, we examined the efficiency of the enzymatic digestion of DNA, that could cause misestimation of 8OHdG. Second, since we considered that the oxygen molecules in atmosphere and in reagents were the main factor contributing to the experimental errors, we carried out the 8OHdG determination under oxygen-free conditions and compared the 8OHdG value with that determined by the methods under ambient atmosphere. The calf thymus DNA was sufficiently digested in the condition we used and the yields of dG were constant, even when the DNA was damaged with H 2 O 2 (80 mM) and UV irradiation. By carrying out the DNA extraction manually, instead of using the DNA extractor, we could reduce the additional 8OHdG formation during sample processing. No trend was found in the difference between the 8OHdG values determined under oxygen-free conditions and under ambient atmosphere. However, when the 8OHdG values were compared in samples with asbestos, the value determined under oxygen-free conditions was significantly lower than that determined under ambient atmosphere. These findings suggest that the removal of oxygen molecules was effective in reducing accidental ROS generation by impurities in the sample, which could cause the additional 8OHdG formation, and that the oxygen-free system made the determination of 8OHdG reproducible and more accurate than before. When the oxygen-free system was applied to human leukocytes, the system showed good reproducibility (r = 0.535, P < 0.001), even though the 8OHdG level was low. With the system, we could detect a significant difference between 8OHdG in polymorphonuclear leukocytes (0.241 ± 0.129) and mononuclear leukocytes (0.188 ± 0.126, P < 0.01).

Background Accidental opening of the frontal sinuses during craniotomy can lead to various postoperative complications. We report a simple and reliable reconstruction method using no exogenous or autogenous material obtained from another site. Methods This method involves packing a small wedge-shaped piece of bone obtained from the bone flap of the craniotomy into the nasal recess of the exposed sinus. The large opened frontal sinus is reconstructed as a new minimized frontal sinus with mucous membrane and bony roof in the nasal recess. Results Eleven patients with large frontal sinus opening during craniotomy (7 cases of bifrontal craniotomy for the basal interhemispheric approach) underwent frontal sinus reconstruction by packing of a small bone piece into the nasal recess. No patients suffered postoperative complications related to the opened frontal sinus such as pneumocephalus or cerebrospinal fluid rhinorrhea. Conclusion Packing of a small bone piece from the bone flap is a quick and reliable method to reconstruct the frontal sinus opened during craniotomy.

Among the various disorders manifesting dementia, gait disturbance, and urinary incontinence in the elderly population, idiopathic normal pressure hydrocephalus (iNPH) is becoming of great importance. After the publication of the first edition of the Guidelines for Management of Idiopathic Normal Pressure Hydrocephalus in 2004 (the English version was published in 2008), clinical awareness of iNPH has risen dramatically, and the number of shunt surgeries has increased rapidly across Japan. Clinical and basic research on iNPH has increased significantly, and more high-level evidence has since been generated. The second edition of the Japanese Guidelines was thus published in July 2011, to provide a series of timely evidence-based recommendations related to iNPH. The revision of the Guidelines has been undertaken by a multidisciplinary expert working group of the Japanese Society of Normal Pressure Hydrocephalus in conjunction with the Japanese Ministry of Health, Labour and Welfare research project on "Studies on the epidemiology, pathophysiology, and treatment of normal pressure hydrocephalus." This English version of the second edition of the Guidelines was made to share these ideas with the international community and to promote international research on iNPH.

The genotoxicity of multi-walled carbon nanotubes (MWCNTs) was evaluated in vivo with comet assays using the lung cells of rats given MWCNTs. The MWCNTs were intratracheally instilled as a single dose at 0.2 or 1.0 mg kg(-1) or a repeated dose at 0.04 or 0.2 mg kg(-1) , once a week for 5 weeks, to male rats. The rats were sacrificed 3 or 24 h after the single instillation and were sacrificed 3 h after the last instillation in the repeated instillation groups. Histopathological examinations of the lungs revealed that MWCNTs caused inflammatory changes including the infiltration of macrophages and neutrophils after a single instillation and repeated instillation at both doses. In comet assays using rat lung cells, no changes in % Tail DNA were found in any group given MWCNTs. These findings indicate that MWCNTs do not have the potential to cause DNA damage in comet assays using the lung cells of rats given MWCNTs at doses causing inflammatory responses.

Mammals, including human beings, have a circadian clock system to regulate behavioral and physiological processes. In this study, we investigated the effect of dosing time on micronucleus induction in the bone marrow by evaluating the frequencies of micronucleated peripheral reticulocytes (MNRETs) in mice exposed to N-ethyl-N-nitrosourea (ENU) to assess any difference in genotoxic sensitivity to chemicals between light and dark periods (inactive phase for rodents and active phase for rodents). Male C3H/He mice were treated intraperitoneally with ENU (12.5 or 25 mg/kg body weight) at zeitgeber time (ZT) 3 in the light period or ZT15 in the dark period, and then the time courses of the frequencies of the MNRETs were determined. The frequencies of the MNRETs induced by ENU increased time-dependently and peaked at 48 hr after treatment for ZT3 and ZT15, and were obviously higher in the ZT15 treatment group than the ZT3 treatment group. The MNRETs were measured at 48 hr after treatment with ENU (25 mg/kg body weight) at various dosing times (ZT0, 3, 6, 12, 15 and 18). The frequencies of the MNRETs in mice treated at ZT0, 15 and 18 were significantly higher than those in mice treated at ZT3, 6 and 12. These results suggest that genotoxic sensitivity to chemicals in mouse bone marrow is different between light and dark periods maybe due to different biological responses (detoxification, cell cycle, DNA repair, etc.) related to circadian rhythms.

The genotoxicity of single-wall carbon nanotubes (SWCNTs) was evaluated in vivo using the comet assay after intratracheal instillation in rats. The SWCNTs were instilled at a dosage of 0.2 or 1.0mg/kg body weight (single instillation group) and 0.04 or 0.2mg/kg body weight once a week for 5weeks (repeated instillation group). As a negative control, 1% Tween 80 was instilled in a similar manner. As a positive control, ethyl methanesulfonate (EMS) at 500mg/kg was administered once orally 3h prior to dissection. Histopathologically, inflammation in the lung was observed for all the SWCNTs in both single and repeated groups. In the comet assay, there was no increase in% tail DNA in any of the SWCNT-treated groups. In the EMS-treated groups, there was a significant increase in% tail DNA compared with the negative control group. The present study indicated that a single intratracheal instillation of SWCNTs (1.0mg/kg) or repeated intratracheal instillation (0.2mg/kg) once a week for five weeks induced a clear inflammatory response (hemorrhage in the alveolus, infiltration of alveolar macrophages and neutrophiles), but no DNA damage, in the lungs in rats. Under the conditions of the test, SWCNTs were not genotoxic in the comet assay following intratracheal instillation in rats.

8-Hydroxydeoxyguanosine (8-OHdG) is a typical form of oxidative DNA damage which causes mutation in vitro and in vivo. We investigated potential factors confounding 8-OHdG determination and, based on the results, then determined the 8-OHdG levels in human peripheral blood leukocytes. 8-OHdG was detected electrochemically after extraction of DNA from the cells without the use of phenol by a DNA extractor under helium. In the preliminary experiments, the mononuclear leukocytes (MN) in blood samples obtained from 19 laboratory workers and students were separated from the polymorphonuclear leukocytes (PMN) with Mono-Poly resolving medium. The 8-OHdG in the MN (1.157 +/- 0.414 molecules per 10(5) deoxyguanosine) did not differ significantly from that in PMN (1.131 +/- 0.418). The effect of red blood cells (RBC) on 8-OHdG formation during DNA extraction was then examined by adding RBC to the human lymphoblastoid cell line FA72. Addition of RBC at ratios of up to 4 RBC per FA72 cell did not increase 8-OHdG levels, while addition at a RBC/FA72 cell ratio of 20 increased the 8-OHdG level 1.43-fold over that without RBC. The potential effect of histidine, a scavenger of both hydroxyl radicals and singlet oxygen, on reduction of artificial 8-OHdG formation during DNA extraction was examined during DNA extraction in the human promyelocytic leukemia cell line HL60. Addition of His decreased the 8-OHdG level dose-dependently (30% reduction at 30 mM His concentration). Based on these results, we determined the 8-OHdG levels in human leukocyte samples obtained from 79 healthy male factory workers aged 24-59 years. The leukocyte fraction containing both MN and PMN was separated from RBC with Mono-Poly resolving medium and DNA was extracted from the leukocytes in the presence of 30 mM His. The mean 8-OHdG level in these samples was 1.072 +/- 0.230. To evaluate the reliability of the assay, FA72 was used as a standard sample in all assay determinations and the 8-OHdG levels of both the leukocyte samples and the FA72 sample(s) were measured in each determination. The inter- and intra-assay coefficients of variation (CV) were calculated to be 14.4% (n = 14) and 3.9-13.5% (n = 3-5 per assay) respectively. The 8-OHdG level was measured twice in 19 leukocyte samples; the value at the first determination was not correlated with that at the second determination. The range of 8-OHdG levels in the samples was relatively small compared with the CV of the assay.(ABSTRACT TRUNCATED AT 400 WORDS)

Polyethylene glycol 400 (PEG 400) is widely used with a variety of pharmaceutical formulations, and is often added to dosing formulations in preclinical toxicity studies. The aim of the present study was to characterize the effects of PEG 400 on the rat gastrointestinal tract. Three dosage levels (5, 50 or 100 v/v%) of PEG 400 were administered at a volume of 5 ml/kg/day by gavage for 15 days to the rats (5 males and 5 females in each group). At the end of the treatment, the whole lengths of gastrointestinal tracts were examined pathologically. Although there were no gross abnormalities at necropsy, the histopathological examination revealed several changes localized to the stomach mucosa, but not in the intestine. The changes consisted of infiltration of eosinophils and globule leukocytes, increased in the height of the entire mucosal layer, elongation of the surface mucous epithelial and mucous neck cell layers with increased intracellular mucous in the glandular stomach, and the spongiosis (intercellular edema) of the squamous epithelium in the forestomach. These changes near the limiting ridge tended to increase in severity and extent in a dose-dependent manner. These results suggest that repeated oral administration of concentrated PEG 400 can easily induce the mucosal changes in the stomach of the rats.

High mobility group box-1 protein (HMGB-1), a protein expressed highly in developing neurons, is involved in the development and differentiation of neurons. At the same time, it functions as a transcriptional regulator of particular genes and as a cytokine: HMGB-1 released from a defective cell has been reported to induce damage to the adjacent cells.With a view to examine the relationship between neuronal damage caused by hydrocephalus and HMGB-1, we analyzed the expression of HMGB-1 in the cerebellum, cerebrum, and hippocampus of 1-day-old congenitally hydrocephalic H-Tx rats.As opposed to nonhydrocephalic H-Tx rats, the hydrocephalic H-Tx rats were observed to show stronger expression of HMGB-1 in the cerebellum, cerebrum, and hippocampus. Consequently, the protein was presumed to influence the development of neurons from an early postnatal stage not only in the cerebral cortex and hippocampus but also in the cerebellum, which is less susceptible to the direct effects of hydrocephalus. We expect that, in the future, regulating the expression or functions of HMGB-1 will lead to the possibility of impeding the progress of neuronal damage caused by hydrocephalus.

We have previously reported that the level of leucine-rich alpha-2-glycoprotein (LRG) expression is specifically increased in cerebrospinal fluid (CSF) of idiopathic normal pressure hydrocephalus (INPH). The objective of this study is to examine the localization of LRG - the cerebral areas where it is expressed.The histological sections of autopsied brain specimens from ten subjects, five adult cases (mean age 43.6 years; range 34-50 years) and five senile cases (mean age 76.0 years; range 67-88 years) were prepared, multistained with antibodies against human LRG, glial fibrillary acidic protein (GFAP), CD31, and aquaporin-4 (AQP4), and reviewed for the expression sites of LRG.Immunostains of GFAP and LRG were compared in standard brain specimens from elderly patients. The results indicated that LRG is distributed throughout the entire brain, with especially high expression in the deep cerebral cortex. In addition, the cells that express LRG showed similar morphology to astrocytes. Double staining of CD31 and LRG revealed a significant expression of LRG in the pericapillary regions. The expression was observed in resident astrocytes, as well as in the capillary vessel to which astrocytic processes grow and adhere. When age-related comparisons were made between senile and adult specimens, LRG expression increased with age.LRG expression in resident astrocytes increased with age.

Neuroendoscopic surgery is distinct from usual craniotomy as it is performed in water. We have previously reported that the use of artificial cerebrospinal fluid (CSF) as perfusate in third ventriculostomy is more efficacious in minimizing severe host reaction than normal saline or lactated Ringer's solution. In this study, we investigated the effects of different perfusion solutions in human cultured astrocytes. We cultured human astrocytes in growth medium. Then each of them was further cultured for 6 h in artificial CSF, lactated Ringer's solution, or normal saline. Using DNA microarray, RNAs were extracted from each of the cells and were comprehensively analyzed to identify differences in patterns of gene manifestation. Compared to the use of artificial CSF, in cases where lactated Ringer's solution or normal saline was used, there was little difference in the pattern of gene manifestation, but there was an increase in gene manifestation related to apoptosis and inflammatory reaction. For neuroendoscopic surgery, the use of artificial CSF as a perfusate is considered effective in maintaining brain homeostasis compared to the use of normal saline or lactated Ringer's solution.

Neuroendoscopic surgery is distinct from usual craniotomy as it is performed in water. We have previously reported that the use of artificial cerebrospinal fluid (CSF) as perfusate in third ventriculostomy is more efficacious in minimizing severe host reaction than normal saline or lactated Ringer's solution. In this study, we investigated the effects of different perfusion solutions in human cultured astrocytes. We cultured human astrocytes in growth medium. Then each of them was further cultured for 6 h in artificial CSF, lactated Ringer's solution, or normal saline. Using DNA microarray, RNAs were extracted from each of the cells and were comprehensively analyzed to identify differences in patterns of gene manifestation. Compared to the use of artificial CSF, in cases where lactated Ringer's solution or normal saline was used, there was little difference in the pattern of gene manifestation, but there was an increase in gene manifestation related to apoptosis and inflammatory reaction. For neuroendoscopic surgery, the use of artificial CSF as a perfusate is considered effective in maintaining brain homeostasis compared to the use of normal saline or lactated Ringer's solution.

Objectives: We have previously reported that the level of leucine-rich alpha-2-glycoprotein (LRG) expression is specifically increased in cerebrospinal fluid (CSF) of idiopathic normal pressure hydrocephalus (INPH). The objective of this study is to examine the localization of LRG – the cerebral areas where it is expressed.

High mobility group box-1 protein (HMGB-1), a protein expressed highly in developing neurons, is involved in the development and differentiation of neurons. At the same time, it functions as a transcriptional regulator of particular genes and as a cytokine: HMGB-1 released from a defective cell has been reported to induce damage to the adjacent cells.

The genotoxicity of fullerene C(60) nanoparticles was evaluated in vivo with comet assays using the lung cells of rats given C(60) nanoparticles. The C(60) nanoparticles were intratracheally instilled as a single dose at 0.5 or 2.5mg/kg or repeated dose at 0.1 or 0.5mg/kg, once a week for 5 weeks, to male rats. The lungs were obtained 3 or 24h after a single instillation and 3h after repeated instillation. Inflammatory responses were observed in the lungs obtained 24h after a single instillation at 2.5mg/kg and repeated instillation at 0.5mg/kg. Histopathological examinations revealed that C(60) nanoparticles caused slight changes including hemorrhages in alveoli and the cellular infiltration of macrophages and neutrophils in alveoli. In comet assays using rat lung cells, no increase in % Tail DNA was found in any group given C(60) nanoparticles. These findings indicate that C(60) nanoparticles had no potential for DNA damage in comet assays using the lungs cells of rats given C(60) even at doses causing inflammation.

Scenario Graph is a structured mind mapping methodology that aids design teams to generate potential scenarios for new products and services while visually organizing contextual information. Since its introduction in industry and academia, the Scenario Graph has helped design teams to capture new values and behaviors of potential customers during the problem formulation stage. At the same time, the Scenario Graph, sharing a common challenge with various design methods, has faced difficulty regarding validation of its effectiveness as a design method. This paper describes a validation framework for a method in problem formulation stages and an experiment, which compare ideation results of 87 people — 43 people with the Scenario Graph method (as a test group) and 44 people with the Brainstorming (as a control group) — to solve an identical problem. While the results show no statistically significant difference in the number of ideas generated, the data reveals statistically significant differences in the quality of ideas. The test group, which used the Scenario Graph, yielded ideas that were more novel, feasible, and abstract than the control group, which used the Brainstorming, did. These metrics represent a way to measure the quality of ideas in the domain of engineering design. Our experiment confirms the hypotheses that the Scenario Graph is effective in improving the performance of idea generation sessions, which is consistent with qualitative evaluations. The lessons, gained from this experiment, provide an insight on how this method can be effectively used during the early stages of concept generation of a company’s process for product and/or service development.

A working group convened at the 2009 5th IWGT to discuss possibilities for improving in vivo genotoxicity assessment by investigating possible links to standard toxicity testing. The working group considered: (1) combination of acute micronucleus (MN) and Comet assays into a single study, (2) integration of MN assays into repeated-dose toxicity (RDT) studies, (3) integration of Comet assays into RDT studies, and (4) requirements for the top dose when integrating genotoxicity measurements into RDT studies. The working group reviewed current requirements for in vivo genotoxicity testing of different chemical product classes and identified opportunities for combination and integration of genotoxicity endpoints for each class. The combination of the acute in vivo MN and Comet assays was considered by the working group to represent a technically feasible and scientifically acceptable alternative to conducting independent assays. Two combination protocols, consisting of either a 3- or a 4-treament protocol, were considered equally acceptable. As the integration of MN assays into RDT studies had already been discussed in detail in previous IWGT meetings, the working group focussed on factors that could affect the results of the integrated MN assay, such as the possible effects of repeated bleeding and the need for early harvests. The working group reached the consensus that repeated bleeding at reasonable volumes is not a critical confounding factor for the MN assay in rats older than 9 weeks of age and that rats bled for toxicokinetic investigations or for other routine toxicological purposes can be used for MN analysis. The working group considered the available data as insufficient to conclude that there is a need for an early sampling point for MN analysis in RDT studies, in addition to the routine determination at terminal sacrifice. Specific scenarios were identified where an additional early sampling can have advantages, e.g., for compounds that exert toxic effects on hematopoiesis, including some aneugens. For the integration of Comet assays into RDT studies, the working group reached the consensus that, based upon the limited amount of data available, integration is scientifically acceptable and that the liver Comet assay can complement the MN assay in blood or bone marrow in detecting in vivo genotoxins. Practical issues need to be considered when conducting an integrated Comet assay study. Freezing of tissue samples for later Comet assay analysis could alleviate logistical problems. However, the working group concluded that freezing of tissue samples can presently not be recommended for routine use, although it was noted that results from some laboratories look promising. Another discussion topic centred around the question as to whether tissue toxicity, which is more likely observed in RDT than in acute toxicity studies, would affect the results of the Comet assay. Based on the available data from in vivo studies, the working group concluded that there are no clear examples where cytotoxicity, by itself, generates increases or decreases in DNA migration. The working group identified the need for a refined guidance on the use and interpretation of cytotoxicity methods used in the Comet assay, as the different methods used generally lead to inconsistent conclusions. Since top doses in RDT studies often are limited by toxicity that occurs only after several doses, the working group discussed whether the sensitivity of integrated genotoxicity studies is reduced under these circumstances. For compounds for which in vitro genotoxicity studies yielded negative results, the working group reached the consensus that integration of in vivo genotoxicity endpoints (typically the MN assay) into RDT studies is generally acceptable. If in vitro genotoxicity results are unavailable or positive, consensus was reached that the maximum tolerated dose (MTD) is acceptable as the top dose in RDT studies in many cases, such as when the RDT study MTD or exposure is close (50% or greater) to an acute study MTD or exposure. Finally, the group agreed that exceptions to this general rule might be acceptable, for example when human exposure is lower than the preclinical exposure by a large margin.

Objective Cerebrospinal fluid (CSF) shunting can improve symptoms of elderly patients' idiopathic normal pressure hydrocephalus (iNPH). However, adjunctive means for confirming the diagnosis remain unavailable. We have previously reported the specific increase of leucine-rich alpha-2-glycoprotein (LRG) in iNPH CSF, and the present study investigates its potential clinical applications. Methods We performed CSF tap test (TT) on 90 patients (mean age 73.4 years) and shunting in 52 patients (mean age 73.5 years), evaluating symptom improvement and higher cerebral functions—mini-mental state examination (MMSE) and Frontal Assessment Battery (FAB) before and 12 months after shunting. LRG and tau protein concentrations in TT CSF were simultaneously measured using enzyme-linked immunosorbent assay. We then compared the predictive value of these concentrations with TT results regarding successful shunting outcomes. Results Positive combinations of TT and LRG concentrations of 67 ng/ml or higher, gave 81.6% sensitivity and 78.6% specificity. Therefore we used LRG (67 ng/ml) and tau (200 pg/ml) cut-off values, dividing patients into four groups. In group A (LRG ≥ 67 ng/ml and tau

Sturge-Weber syndrome (SWS) is a rare neurocutaneous syndrome characterized by intracranial leptomeningeal angioma, facial port-wine nevi, and glaucoma. Diagnosis is relatively easy because of the facial angioma and MRI findings, but evaluating severity is difficult. Predictors of SWS's prognosis are epilepsy and brain dysfunction under the leptomeningeal angioma. Therefore, active research has been intensely conducted with electrophysiological, neuroimaging, and neuropsychological methods. Final goals of this research are to define the therapeutic strategy. In this review, we focus on recent advances in neuroimaging and EEG analysis to discover the epileptogenesis, the most adequate therapy, and prospective topics of investigation in SWS.

The Non-genotoxic Carcinogen Study Group in the Environmental Mutagen Society of Japan organised the second step of the inter-laboratory collaborative study on one-stage and two-stage cell transformation assays employing BALB/c 3T3 cells, with the objective of confirming whether the respective laboratories could independently produce results relevant to initiation or promotion. The method was modified to use a medium consisting of DMEM/F12 supplemented with 2% fetal bovine serum and a mixture of insulin, transferrin, ethanolamine and sodium selenite, at the stationary phase of cell growth. Seventeen laboratories collaborated in this study, and each chemical was tested by three to five laboratories. Comparison between the one-stage and two-stage assays revealed that the latter method would be beneficial in the screening of chemicals. In the test for initiating activity with the two-stage assay (post-treated with 0.1μg/ml 12- O-tetradecanoylphorbol-13-acetate), the relevant test laboratories all obtained positive results for benzo[ a]pyrene and methylmethane sulphonate, and negative results for phenanthrene. Of those laboratories assigned phenacetin for the initiation phase, two returned positive results and two returned negative results, where the latter laboratories tested up to one dose lower than the maximum dose used by the former laboratories. In the exploration of promoting activity with the two-stage assay (pretreated with 0.2μg/ml 3-methylcholanthrene), the relevant test laboratories obtained positive results for mezerein, sodium orthovanadate and TGF-β1, and negative results for anthralin, phenacetin and phorbol. Two results returned for phorbol 12,13-didecanoate were positive, but one result was negative — again, the maximum dose to achieve the latter result was lower than that which produced the former results. These results suggest that this modified assay method is relevant, reproducible and transferable, provided that dosing issues, such as the determination of the maximum dose, are adequately considered. The application of this two-stage assay for screening the initiating and promoting potential of chemicals is recommended for consideration by other research groups and regulatory authorities.

The authors have developed a minimally invasive lumboperitoneal shunt placement procedure conducted after administration of a local anesthetic. The procedure involves placing a guide wire and a peel-away sheath under fluoroscopic and CT guidance. Between June 2004 and August 2006, 40 patients (21 men and 19 women; mean age 72.5 years [range 33-86 years]) underwent surgery. A Codman Hakim programmable valve system (82-3844, CodmanShurtleff, Inc.) was used for the procedure. The mean operating time was 53 minutes, and 7 patients (17.5%) developed shunt dysfunction complications. These complications comprised an infected shunt valve in 2 patients, postoperative lower-limb pain in 1 patient, and shunt obstruction (caused by debris and hemorrhage) at the ventral and lumbar ends in 2 patients each. This procedure is less invasive than conventional lumboperitoneal shunt insertion and could be performed as an outpatient surgery for treatment of idiopathic normal-pressure hydrocephalus.

A single-dose oral toxicity lethal-dose study was conducted to examine the toxicity of capsinoids contained in CH-19 Sweet extract. CH-19 Sweet extract was administered once by gavage to SPF (Crl:CD(SD)) Sprague-Dawley male and female rats at dose levels of 0 (vehicle), 5, 10, or 20 ml/kg of body weight (BW). The concentration of capsinoids in the CH-19 Sweet extract was 71.25 mg/ml; this resulted in administered dose levels of capsinoids of 356.25, 712.5, and 1425 mg/kg BW, respectively. The toxicity of CH-19 Sweet extract by single oral administration was low; only transient salivation or decreased spontaneous movement was observed on the day of administration at > or =10 ml/kg BW. It was concluded that the lethal dose of CH-19 Sweet extract was estimated to be higher than 20 ml/kg (1425 mg/kg as capsinoids) for both males and females since no deaths were observed at any dose in this study. A bacterial reverse mutation test of CH-19 Sweet extract was performed employing Salmonella typhimurium and Escherichia coli and using the preincubation method. Treatment with CH-19 Sweet extract did not increase the number of revertant colonies compared with negative controls either in the presence (+S9) or absence (-S9) of metabolic activation. An in vitro chromosome aberration test was conducted using Chinese hamster lung cultured cells (CHL/IU). Treatment with CH-19 Sweet extract failed to induce chromosome aberrations in either short-term or continuous treatment scenarios, with or without metabolic activation (-S9, +S9). In an in vivo micronucleus test using BDF(1) male mice, CH-19 Sweet extract failed to increase the incidence of micronucleated polychromatic erythrocytes (MNPCEs) or decrease the ratio of polychromatic erythrocytes (PCEs) in any of the treatment groups. These results suggest the absence of mutagenicity as well as in vitro and in vivo clastogenicity of capsinoids contained in CH-19 Sweet extract.

We examined the dose-dependency of gene expression changes for 51 genes in mouse liver treated with two N-nitroso genotoxic hepatocarcinogens, diethylnitrosamine (DEN) and ethylnitrosourea (ENU) by quantitative real-time PCR (qPCR). DEN (3, 9, 27 and 80mg/kg bw) or ENU (6, 17, 50 and 150mg/kg bw) was injected intraperitoneally into groups of five male 9-week-old B6C3F(1) mice and the livers were dissected after 4h and 28 days. Total RNA from pooled livers was reverse-transcribed to cDNA and the amount of each gene was quantified by qPCR. Results were analyzed by hierarchical and k-means clustering and ingenuity pathway analysis (IPA). The most characteristic result was a similar dose-dependency of gene expression changes with DEN and ENU. Twenty-one genes exhibited a distinct dose-dependent increase in expression at 4h for both carcinogens [Bax, Btg2, Ccng1, Cdkn1a, Cyp4a10, Cyp21a1, Fos, Gadd45b, Gdf15, Hmox1, Hspb1, Isg20l1, Jun, Mbd1, Mdm2, Myc, Net1, Plk2, Ppp1r3c, Rcan1 and Tubb2c], although the increase in gene expression due to ENU was generally weaker than that due to DEN. Only Gdf15 showed a dose-dependent increase in expression at 28 days for both carcinogens. The differences between DEN and ENU were in the expression of additional genes (7 for DEN and 8 for ENU). IPA extracted five gene networks: Network-1 included genes related to cancer and cell cycle arrest and associated with Bax, Btg2, Ccng1, Cdkn1a, Gadd45b, Gdf15, Hspb1, Mdm2 and Plk2 and Network-2 was related to DNA replication, recombination, repair and cell death and associated with Cyp21a1, Gdf15, Ppp1r3c, Rcan1 and Tubb2c. The present results show a distinct dose-dependency of gene expression changes induced by DEN and ENU. These changes were associated with cancer, cell cycle arrest, DNA replication, recombination, repair and cell death and were seen not only at 4h but also, for some, at 28 days after administration.

The mechanism of epilepsy in Sturge–Weber syndrome (SWS) has not been elucidated, since it is difficult to detect epileptic discharges on scalp EEG records. To determine the electrophysiological characteristics of SWS, we evaluated epileptic discharges using intracranial electrocorticography (ECoG). We report a 1-year and 3-month-old boy diagnosed with SWS whose psychomotor development was impaired despite good control of his seizures with several antiepileptic drugs (AEDs). Prolonged video ECoG monitoring was performed to determine whether subclinical seizures had induced his developmental delay. Frequent seizures were detected as motionless staring and respiratory distress. His seizures originated from multiple areas along the Sylvian fissure, propagated very slowly, and lasted for 10 to 30 min. Left hemispherotomy was performed, and he was confirmed to be completely seizure-free after surgery. Our findings indicate that long-lasting and barely discernible seizures may continue in patients with SWS who appear to have good control of epilepsy with AEDs.

The roadable aircraft which has an inflatable wing has been investigated by the authors. In this research, guillotine test was conducted first in order to substantiate a factor of safety 2 for an inflatable membrane. Then, structural analyses of a modified inflatable wing, for three cases, internal pressure only, with 3.8G maneuver load, and with lateral gust load, were performed to predict deformation and stress on an inflatable wing. Finally, the aerodynamic characteristics of a wing and the influence on its characteristics by the deformation of a wing are investigated by CFD simulations and wind tunnel test.

A film-type multipoint pressure sensor was developed by using an optical fiber with the Fiber Bragg Grating (FBG), which enable to measure the pressure only by sticking the sensor on a body surface. In this paper, the developed pressure sensor and the pressure measurement system are described. In order to clarify the performance of the FBG pressure sensor, pressure distributions on NACA4412 airfoil model were measured in wind tunnel test. The measured pressure was compared with CFD simulation results and other experiment data. It was found that the pressure variation on the wing surface was well measured by the developed FBG pressure sensor both in positive and negative pressure sides for various attack angles.

Some cases of compensatory hydrocephalus have been reported in which cognitive deficiency progresses despite the absence of progressive ventricular dilation. In this study, the differentially expressed genes in compensated hydrocephalic H-Tx rat cortices were determined. A molecular mechanism that induces neuronal death in the cerebral cortex of compensated hydrocephalus is proposed.The cerebral cortices of 8-week-old H-Tx rats with spontaneously arrested hydrocephalus (hH-Tx) and nonhydrocephalic H-Tx (nH-Tx) control rats were subjected to cDNA microarray analysis followed by canonical pathway analysis.In the hH-Tx rats, many genes in the amyloidal processing pathway showed altered expression, including Akt3 and p38 MAPK. These latter genes are involved in tau protein phosphorylation, and their increased expression in hydrocephalus was confirmed by real-time polymerase chain reaction analysis. Immunohistological and immunoblot analysis revealed elevated phosphorylated tau expression in the cerebral cortex neurons of the hH-Tx rats.The accumulation of phosphorylated tau protein in the cerebral cortex may be one of the mechanisms by which later cognitive dysfunction develops in patients with compensated hydrocephalus. More work needs to be done to determine if the accumulation of phosphorylated tau in the cortex can help predict which patients may decompensate thus requiring more aggressive treatment for compensated hydrocephalus.

The present authors have been investigated a roadable aircraft with an inflatable ring wing as for the future aircraft. In this research, the displacement of ring wing and stress on wing pressured internally and exerted ultimate load are simulated by using non-linear FEM cord, MARC. Furthermore, wind tunnel tests were conducted to predict the influence of a deformation of wing on aerodynamic characteristics. It is clarified that a ring wing can keep its shape if ribs which can stand compressive stress set inside, and the deformation of airfoil at internal pressure 1atm does not affect on aerodynamic characteristics.

Accumulating evidence suggests that reticulocytes (RETs) in the peripheral blood of rats may represent a suitable cell population for use in the micronucleus assay, despite the ability of the rat spleen to selectively remove micronucleated erythrocytes from the peripheral circulation. To evaluate the analytical performance of a previously described flow cytometric method (Torous et al., 2003, Toxicol. Sci. 74, 309-314) that may allow this assay to be conducted using peripheral blood in lieu of bone marrow sampling, we compared the sensitivity and performance characteristics of the flow cytometric technique with two established microscopy-based scoring methods. Peripheral blood samples from single Sprague-Dawley rats treated for 6 days with either vehicle or cyclophosphamide were prepared in replicate for scoring by the three methods at different laboratories. These blood-based measurements were compared to those derived from bone marrow specimens from the same animals, stained with acridine orange, and scored by microscopy. Through the analysis of replicate specimens, inter- and intralaboratory variability were evaluated for each method. Scoring reproducibility over time was also evaluated. These data support the premise that rat RETs harvested from peripheral blood are a suitable cell population to assess genotoxicant-induced micronucleus formation. The interlaboratory comparison provides evidence of the general robustness of the micronucleus endpoint using different analytical approaches. Furthermore, data presented herein demonstrate a clear advantage of flow cytometry-based scoring over microscopy-significantly lower inter- and intralaboratory variation and higher statistical sensitivity.

As for a possible future vehicle, several types of roadable aircrafts have been studied recently. In this paper, a compact type roadable aircraft with a fixed ring wing is investigated. When a roadable aircraft is driven on road, its dimension has to be restricted by the Road Traffic Act, which indicates that the wing cannot be large. In flight, an enough wing size is needed to keep the weight and takeoff. One possible solution to solve this antinomy is an inflatable wing. In order to prove a possibility of an inflatable ring wing, the deformation of a ring wing and the aerodynamics characteristics are investigated. The FEM simulations are performed to examine the deformation of an inflatable ring wing by using ANSYS and MARC. The influences of deformations of a ring wing upon lift and drag are clarified by numerical simulation with the use of a commercial CFD code, SCRYU/Tetra.