Genistein and menaquinone-4 treatment-induced alterations in the expression of mRNAs and their products are beneficial to osteoblastic MC3T3-E1 cell functions.

The aim of the present study was to determine the molecular basis of the beneficial effects of genistein and/or menaquinone­4 (MK­4) on bone quality. Initially, 1 µM genistein was applied to MC3T3­E1 cells for 24 h and the upregulated mRNAs that were detected by microarray were selected for further examination by reverse transcription­quantitative­polymerase chain reaction. Among them, alterations were observed in the level of GATA­binding protein 6 (GATA6), Notch gene homolog 2 (NOTCH2), Wnt family member 5A (WNT5A), bone γ­carboxyglutamate protein (BGLAP), chondroadherin (CHAD), dipeptidyl peptidase 4 (DPP4), ectonucleotide pyrophosphatase/phosphodiesterase 2 (ENPP2), alkaline phosphatase (ALP) 3 and ATPase phospholipid­transporting 11A (ATP11A) in response to treatment with 0.1 µM 17­ß­estradiol, 1 µM genistein, and/or 1 µM MK­4. GATA6, NOTCH2 and WNT5A are considered to be associated with osteoclast, but not osteoblast, function; however, increases in osteoblastic mRNAs, including BGLAP and CHAD, were observed in each of the treatment groups at 48 h. Immunocytochemical analysis confirmed an increase in CHAD and DPP4 proteins following the administration of genistein + MK­4. Furthermore, genistein + MK­4 led to alterations in cell morphology to spindle or oval shapes, and increased the intensity of ALP staining. Although the level of ALP mRNA was not consistently altered in response to the treatments, a marked increase in ALP activity was observed following 96 h treatment with genistein + MK­4. Therefore, the simultaneous intake of genistein and MK­4 appears to be beneficial for the maintenance of bone quality.

Allyl nitrile: Toxicity and health effects.

OBJECTIVES: Allyl nitrile (3-butenenitrile) occurs naturally in the environment, in particular, in cruciferous vegetables, indicating a possible daily intake of the compound. There is no report on actual health effects of allyl nitrile in humans, although it is possible that individuals in the environment are at a risk of exposure to allyl nitrile. However, little is known about its quantitative assessment for the environment and bioactivity in the body. This study provides a review of previous accumulated studies on allyl nitrile. METHODS: Published literature on allyl nitrile was examined for findings on toxicity, metabolism, risk of various cancers, generation, intake estimates, and low-dose effects in the body. RESULTS: High doses of allyl nitrile produce toxicity characterized by behavioral abnormalities, which are considered to be produced by an active metabolite, 3,4-epoxybutyronitrile. Cruciferous vegetables have been shown to have a potential role in reducing various cancers. Hydrolysis of the glucosinolate sinigrin, rich in cruciferous vegetables, results in the generation of allyl nitrile. An intake of allyl nitrile is estimated at 0.12 µmol/kg body weight in Japan. Repeated exposure to low doses of allyl nitrile upregulates antioxidant/phase II enzymes in various tissues; this may contribute to a reduction in neurotoxicity and skin inflammation. These high and low doses are far more than the intake estimate. CONCLUSION: Allyl nitrile in the environment is a compound with diverse bioactivities in the body, characterized by inducing behavioral abnormalities at high doses and some antioxidant/phase II enzymes at low doses.

Anti-Inflammatory and Antioxidant Effects of Repeated Exposure to Cruciferous Allyl Nitrile in Sensitizer-Induced Ear Edema in Mice.

BACKGROUND: Skin sensitizers induce allergic reactions through the induction of reactive oxygen species. Allyl nitrile from cruciferous vegetables has been reported to induce antioxidants and phase II detoxification enzymes in various tissues. We assessed the effects of repeated exposure to allyl nitrile on sensitizer-induced allergic reactions. MATERIAL AND METHODS: Mice were dosed with allyl nitrile (0-200 µmol/kg), and then received a dermal application of 1 of 3 sensitizers on the left ear or 1 of 2 vehicles on the right ear. Quantitative assessment of edema was carried out by measuring the difference in weight between the portions taken from the right and left ears. We tested enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and thiobarbituric acid reactive substances (TBARS) in ears. RESULTS: Repeated exposure to allyl nitrile reduced edemas induced by glutaraldehyde and by 2, 4-dinitrochlorobenzene (DNCB), but not by formaldehyde. The repeated exposure decreased levels of TBARS, a marker of oxidative stress, induced by glutaraldehyde and by DNCB, but not by formaldehyde. Allyl nitrile elevated SOD levels for the 3 sensitizers, and CAT levels for formaldehyde and DNCB. Allyl nitrile also increased GPx levels for formaldehyde and DNCB, but not for glutaraldehyde. The reduced edemas were associated with changes in oxidative stress levels and antioxidant enzymes. CONCLUSIONS: Repeated exposure to allyl nitrile reduced allergic reactions induced by glutaraldehyde and by DNCB, but not by formaldehyde. This reduction was associated with changes in ROS levels and antioxidant enzyme activities.

Factors associated with tuberculosis cases in Semarang District, Indonesia: case-control study performed in the area where case detection rate was extremely low.

OBJECTIVES: Indonesia is ranked as the 4th highest contributor to tuberculosis (TB) in the world. Semarang District in Central Java displays extremely low case detection rate (CDR), possibly contributing to the local prevalence of TB. METHODS: A case-control study was performed to explore the factors that cause such low CDR. We recruited 129 TB cases and 83 controls that visited the same centers and were not diagnosed with TB. RESULTS: The cases had 7.5 ± 2.3 symptoms/person on average, indicating the delay in diagnosis because the controls only displayed 1.0 ± 1.7. The multiple logistic regression analysis comparing the cases/controls extracted following factors as a risk to have TB: farmer, close contact with TB patients, ignorance of whether Bacillus Calmette-Guérin (BCG) was accepted or no, smoking, low income, a lot of people living in the same room, irregular hand wash before meals, not wash hands after blow, soil floor, and no sunlight and no ventilation in the house. CONCLUSIONS: Neither the cases nor the controls knew the symptoms and how to avoid TB infection, which probably caused the delay in diagnosis. It is difficult to change the current living conditions. Thus, the amendment of the community-based education program of TB seems to be required.

Regulation of serine protease inhibitor Kazal type-5 (SPINK5) gene expression in the keratinocytes.

OBJECTIVES: Serine protease inhibitor Kazal type-5 (SPINK5) plays a crucial role in deciding the timing of desquamation of the skin. Its gene expression is limited at the very surface of the stratum granulosum (SG), whereas expression of kallikreins (KLKs) encoding proteases is usually found throughout the stratum spinosum and SG. METHODS: To explore the difference in expression regulation of these proteases/inhibitors, the function of SPINK5 promoter was examined using luciferase assay. RESULTS: Luciferase assay targeting the SPINK5 promoters (nucleotide -676/-532 and -318/-146 from the major transcription start site) showed high intensity in NHEK human keratinocyte. These two sites had neither common cis-elements nor GATA3 element but electrophoretic mobility shift assay showed similar retardation bands. Moreover, DNA footprinting did not display specific protected bands. Thus, we could not identify cis-element(s) that controlled these elements. Differentiation induced by high Ca(2+) medium failed to alter their luciferase activities. Transfection of GATA3 expressing vector significantly but slightly increased them and that of vector expressing its dominant negative form decreased. CONCLUSIONS: Although GATA3 is reportedly important for inhibition of proliferation and induction of differentiation of keratinocytes, its effect on SPINK5 expression was indirect and GATA3 alone was insufficient for final differentiation of keratinocytes where full SPINK5 expression was observed.

Posttraumatic stress disorders in the Nanai after pollution of the Amur River: ethnocultural analysis.

OBJECTIVES: Chemical pollution of the Amur River has seriously damaged traditions and caused posttraumatic stress disorder (PTSD) among the Nanai, the indigenous people living along this river. This study was performed to clarify the ethnographic characteristics of PTSD in this unique population. METHODS: The study group consisted of 75 male and 112 female randomly selected volunteers. PTSD severity measured using scores of the Impact of Event Scale--Revised (Total-I) and Clinical-Administered PTSD Scale (Total-C) was estimated according to demographic and ethnocultural backgrounds, clinical status, and ethnopsychological attitudes toward the Amur River. RESULTS: The differences in averages of Total-I and Total-C were not always the same in the groups divided by ethnographic information. Logistic regression analysis with a dependent variable, possibly without PTSD (Total-I <34 and Total-C <40)/possibly with PTSD (either Total-I ≥34 or Total-C ≥40), and categorical independent variables using ethnographic information extracted a low score when 'priority values' and 'the Amur River for me is' was "profession" and a high score when 'dominant role in spousal relationship' was "self," when 'predominant forms of response in stressful situations' was "try to organize," when 'preferred method of medical treatment' was specific for the Nanai, when "rely on something mystical" was manifested, and when the Amur River was believed to be "sacred". CONCLUSION: Those with a pragmatic attitude were less likely to have PTSD. However, those who were required to make decisions within close relationships and were intimate with the Nanai tradition and the Amur River had increased likelihood of PTSD.

A distinct effect of transient and sustained upregulation of cellular factor XIII in the goldfish retina and optic nerve on optic nerve regeneration.

Unlike in mammals, fish retinal ganglion cells (RGCs) have a capacity to repair their axons even after optic nerve transection. In our previous study, we isolated a tissue type transglutaminase (TG) from axotomized goldfish retina. The levels of retinal TG (TG(R)) mRNA increased in RGCs 1-6weeks after nerve injury to promote optic nerve regeneration both in vitro and in vivo. In the present study, we screened other types of TG using specific FITC-labeled substrate peptides to elucidate the implications for optic nerve regeneration. This screening showed that the activity of only cellular coagulation factor XIII (cFXIII) was increased in goldfish optic nerves just after nerve injury. We therefore cloned a full-length cDNA clone of FXIII A subunit (FXIII-A) and studied temporal changes of FXIII-A expression in goldfish optic nerve and retina during regeneration. FXIII-A mRNA was initially detected at the crush site of the optic nerve 1h after injury; it was further observed in the optic nerve and achieved sustained long-term expression (1-40days after nerve injury). The cells producing FXIII-A were astrocytes/microglial cells in the optic nerve. By contrast, the expression of FXIII-A mRNA and protein was upregulated in RGCs for a shorter time (3-10days after nerve injury). Overexpression of FXIII-A in RGCs achieved by lipofection induced significant neurite outgrowth from unprimed retina, but not from primed retina with pretreatment of nerve injury. Addition of extracts of optic nerves with injury induced significant neurite outgrowth from primed retina, but not from unprimed retina without pretreatment of nerve injury. The transient increase of cFXIII in RGCs promotes neurite sprouting from injured RGCs, whereas the sustained increase of cFXIII in optic nerves facilitates neurite elongation from regrowing axons.

Inpatient satisfaction and job satisfaction/stress of medical workers in a hospital with the 7:1 nursing care system (in which 1 nurse cares for 7 patients at a time).

OBJECTIVES: Inpatient satisfaction, job satisfaction/stress of medical workers, and hospital profitability under the 7:1 nursing care system (in which 1 nurse cares for 7 patients at a time) were compared with those under the 10:1 system at a hospital with the diagnosis procedure combination (DPC) payment system. METHODS: A total of 202 inpatients discharged from the Departments of Cardiology and Metabolism completed an inpatient satisfaction questionnaire. A total of 108 medical workers were recruited to survey their job satisfaction/stress and to estimate the effects of the DPC. The profits for 10 cardiac and metabolic diseases in 2008 were compared with those in 2007. RESULTS: Mean inpatient satisfaction scores were around 4 ("somewhat satisfied") under both the 10:1 and 7:1 systems, and increased significantly to 4.14-4.38 under the 7:1 system. Excluding workload of physicians, the other stresses of physicians/nurses remained unaltered, as did their low job satisfaction. They estimated their understanding of the DPC as insufficient but felt that introducing the DPC neither shortened length of stay nor improved "the quality of medical/nursing care," regardless of the system. Total percentage profit in 2008 was almost the same as that in 2007, whereas diseases with deficits increased from 3 to 4. [corrected] CONCLUSIONS: The 7:1 system was somewhat beneficial for inpatients but not always for medical worker quality of life (QOL) or for hospital income, which are important to maintain high quality of medical/nursing care. It is important to further explore factors increasing QOL of medical workers and hospital income.

Abundant expression of Kallikrein 1 gene in human keratinocytes was mediated by GATA3.

Among Tissue kallikrein genes (KLKs), KLK1 is abundantly expressed in human skin. Although its putative promoter is known to have various cis-elements, they have not been functionally tested. In the present study, the regulation mechanism of KLK1 promoter supporting such abundant expression was examined. Luciferase assay targeting the KLK1 promoter (nucleotide -1153/+40 from the major transcriptional start site) was performed on NHEK human keratinocyte. -954/-855, -428/-236, and -100/+40 had the induction activity. The motif search program failed to find unique binding motifs in -428/-236, whereas both -954/-855 and -100/+40 had a unique GATAs binding motif. Electrophoretic mobility shift assay (EMSA) and DNA footprinting confirmed the binding of NHEK nuclear protein to these motifs that were supershifted by anti-GATA3 antibody. Among GATA isoforms, GATA3 alone could be amplified in RNA obtained from NHEK. Moreover, introduction of GATA3 into fibroblastic NIH3T3 cells enhanced the activity of KLK1 promoter containing -954/+40, while that of GATA3 dominant negative mutant to NHEK cells impaired the same promoter's activity. Thus, GATA3 was found to bind the site located at -954/-855 and to be a key regulator of abundant KLK1 expression in human keratinocyte.

Polymorphisms in the promoter region of the human class II alcohol dehydrogenase (ADH4) gene affect both transcriptional activity and ethanol metabolism in Japanese subjects.

Class II alcohol dehydrogenase (pi-ADH), encoded by alcohol dehydrogenase (ADH4), is considered to contribute to ethanol (EtOH) oxidation in the liver at high concentration. Four single nucleotide polymorphisms (SNPs) were found in the promoter region of this gene. Analysis of genotype distribution in 102 unrelated Japanese subjects revealed that four loci were in strong linkage disequilibrium and could be classified into three haplotypes. The effects of these polymorphisms on transcriptional activity were investigated in HepG2 cells. Transcriptional activity was significantly higher in cells with the -136A allele than in those with the -136C allele. To investigate whether this difference in transcriptional activity caused a difference in EtOH elimination, previous data on blood EtOH changes after 0.4 g/kg body weight alcohol ingestion were analyzed. When analyzed based on aldehyde dehydrogenase-2 gene (ALDH2) (487)Glu/Lys genotype, the significantly lower level of EtOH at peak in subjects with -136C/A and -136A/A genotype compared with subjects with -136C/C genotype indicated that -136 bp was a suggestive locus for differences in EtOH oxidation. This effect was observed only in subjects with ALDH2 (487)Glu/Glu. These results suggested that the SNP at -136bp in the ADH4 promoter had an effect on transcriptional regulation, and that the higher activity of the -136A allele compared with the -136C allele caused a lower level of blood EtOH after alcohol ingestion; that is, individuals with the -136A allele may consume more EtOH and might have a higher risk for development of alcohol dependence than those without the -136A allele.

A promoter polymorphism in the ALDH2 gene affects its basal and acetaldehyde/ethanol-induced gene expression in human peripheral blood leukocytes and HepG2 cells.

AIMS: To assess the effect of the -360G/A polymorphism in the promoter region of the human aldehyde dehydrogenase-2 (ALDH2) gene on its transcription, basal and acetaldehyde/ethanol-induced gene expression was examined by in vivo and in vitro experiments. METHODS: Human peripheral blood leukocytes were collected before and after alcohol ingestion (0.4 g/kg body weight) in 21 healthy young Japanese volunteers with a deficient phenotype of ALDH2 ((487)Glu/Lys), and the levels of ALDH2 mRNA were quantified by real-time RT-PCR. The transcriptional activity of the ALDH2 promoter was investigated by a reporter assay using HepG2 cells in the presence or absence of acetaldehyde/ethanol. RESULTS: The basal level of ALDH2 mRNA was significantly higher in -360A heterozygous subjects than in -360G homozygous subjects. In all subjects, regardless of the genotype, ALDH2 mRNA increased following ethanol ingestion. The promoter activity of a reporter plasmid for -360G was significantly lower than that of a reporter plasmid for -360A. Exposure to acetaldehyde induced a significant increase in the transcriptional activity of the -360G reporter, but not the -360A reporter. CONCLUSIONS: In vivo and in vitro experiments showed that the -360G allele has lower basal transcriptional activity than the -360A allele, whereas acetaldehyde/ethanol-induced gene expression, in general, seems to be more enhanced in individuals homozygous for the -360G allele than in those with the -360A allele. Thus, the promoter polymorphism may be involved in individual differences in acetaldehyde elimination.

A novel neuroprotective role of a small peptide from flesh fly, 5-S-GAD in the rat retina in vivo.

N-beta-Alanyl-5-S-glutathionyl-3,4-dihydroxyphenylalanine (5-S-GAD), an antibacterial substance isolated from flesh fly, has been described as having multipotential biological activities toward various tissues. However, there has been no report testing its action on neural cells. In the present study, we investigate whether 5-S-GAD is neurotoxic or neuroprotective to the rat retina. 5-S-GAD at high doses (more than 200 pmol) induced apoptosis of retinal neurons 7 days after intraocular injection. 5-S-GAD at low doses (2-20 pmol) significantly attenuated the loss of retinal ganglion cells (RGCs) and the thinning of inner retina induced by NMDA in a dose-dependent manner. To understand the protective mechanism of 5-S-GAD, we investigated the influence of 5-S-GAD on the cell survival molecules, phospho-Akt and Bcl-2. 5-S-GAD (2-20 pmol) rapidly increased phospho-Akt expression 1-7 days and Bcl-2 expression 3-7 days after injection. The cellular localization of this increase was both in bipolar cells and RGCs. This neurosurvival effect of 5-S-GAD was further tested using another model of optic nerve injury. 5-S-GAD significantly blocked the apoptosis of RGCs 7 days after optic nerve crush. These results show that 5-S-GAD (2-20 pmol) protects against the NMDA- and optic nerve crush-induced apoptosis of RGCs. The neuroprotective action of 5-S-GAD in the retina might be mediated by the cell survival phospho-Akt/Bcl-2 system and offers a therapeutic option to rescue RGCs from various types of excitotoxic disease, such as glaucoma.

Effects of cruciferous allyl nitrile on phase 2 antioxidant and detoxification enzymes.

BACKGROUND: High intake of cruciferous vegetables has been associated with lower risk of various cancers, and the cancer preventive effect of the vegetables has been associated with their high levels of glucosinolates. The hydrolysis of glucosinolates results in the generation of bioactive compounds, including allyl nitrile which we previously found to be an active inducer of some phase 2 enzymes. In this study, we further explored the inductive ability of this nitrile in light of increasing evidence that antioxidants delay or prevent the development of cancer. MATERIAL/METHODS: Groups of 6 mice were administered subtoxic doses of allyl nitrile (5, 50, 100 or 200 micromol/kg) or vehicle-distilled water daily for 5 days by gastric intubation. On the sixth day, the animals were sacrificed for examination of enzyme activities in their tissues. Enzymes tested were thioredoxin reductase (TR), glutathione peroxidase (GPx), glutathione reductase (GR) and catalase. RESULTS: Allyl nitrile increased the activity of TR in the liver, kidneys and rectum at doses of 100-200 micromol/kg/day, and GPx in the kidneys and small intestine at 50 to 200 micromol/kg/day, while in the colon, alone, it decreased the activities of GR and catalase at doses of 200 and 100-200 micromol/kg/day, respectively not. CONCLUSIONS: The present results suggest an involvement of allyl nitrile in antioxidant defense in the body, except for the colon.

Effects of functional polymorphisms related to catecholaminergic systems on changes in blood catecholamine and cardiovascular measures after alcohol ingestion in the Japanese population.

BACKGROUND: The polymorphism of human aldehyde dehyrogenase-2 (ALDH2) Glu(487)Lys is well known to be a crucial factor underlying the genetic background for alcohol sensitivity in Asian populations. Subjects with the inactive Lys(487) allele show a marked increase in blood acetaldehyde level after alcohol intake, which results in facial flushing and various cardiovascular-related symptoms. However, other polymorphisms related to catecholaminergic systems that tightly regulate the activity of the sympathetic nervous system may also influence the physiological changes after acute alcohol intake. METHODS: We investigated whether, together with the ALDH2 Gly(487)Lys and ADH1B Arg(47)His genotype, putative functionally important polymorphisms, including 9 loci in 7 human genes, were associated with changes in blood catecholamine levels and cardiovascular measures after alcohol ingestion. Forty-nine young Japanese males were subjected to blood catecholamine analysis after alcohol ingestion. Among them, 28 were also subjected to heart rate variability and blood pressure analysis. The contribution of polymorphisms to the alcohol-induced response was analyzed by multiple regression analysis. RESULTS: Among the polymorphisms examined in this study, haplotypes of the phenylethanolamine N-methyltransferase (PNMT) promoter [(-182bpG/A)_(-387bpG/A)] and catechol-O-methyltransferase (COMT) exon 4 [(Ex4 + 119bpC/G)_(Ex4 + 138bpG/A), Leu(136)Leu_Val(158)Met] are suggested to have functionally important effects on alcohol-induced cardiovascular symptoms by affecting blood catecholamine levels. The neuropeptide Y (NPY) promoter C-1450T genotype is also suggested to be involved in the individual differences in regulation of catecholamine secretion. CONCLUSIONS: This study suggested that these common polymorphisms of genes related to catecholaminergic systems, as well as those of the alcohol metabolizing system, are significant for understanding the basis of individual differences in alcohol sensitivity.

Reassessment of the cutoff values of waist circumference and visceral fat area for identifying Japanese subjects at risk for the metabolic syndrome.

In the new world-wide criteria for metabolic syndrome (MetS) by the International Diabetes Federation (IDF) in 2006, the Japanese is the only ethnicity in which the recommended waist circumference (WC) cutoff value is higher in women (>or=90cm) than in men (>or=85cm), and its validity appears to be controversial. We investigated the optimal cutoff points for the diagnosis of central obesity in Japanese men and women, using the receiver operating characteristic (ROC) curve analysis for both of WC and visceral fat area (VFA) in 1870 middle-aged Japanese. VFA was superior to WC and Body mass index (BMI) for discriminating the subjects with two or more nonadipose components of MetS. The optimal cutoff points of VFA and WC were 132.6cm(2) and 89.8cm for men and 91.5cm(2) and 82.3cm for women. The stratifications of MetS components more than 1.0 in average occurred more steeply by the accumulation of VFA in women than in men. In conclusion, setting the cutoff points of WC and VFA lower values in women than in men for the definition of central obesity is needed to identify the subjects with MetS in Japanese, as in other Asian populations.

Abundant expression of nucleosome assembly protein 1 (NAP1) gene in goldfish scale with lateral line.

To explore the possible utilization of goldfish scale in monitoring environmental toxicants as well as that for calcified tissue research, characteristics of cells in the scale were examined by identifying genes differentially expressed in those cells. Subtraction cloning was subjected to RNAs between scale-gill (SG), gill-scale (GS) and scale with lateral line-scale without lateral line (LS). Total numbers of 4, 6, and 9 clones were isolated respectively from SG, GS and LS pools. Blast search of their sequence showed only low homology to other fish sequences so that 5' rapid amplification of cDNA (5'RACE) was applied to the obtained 5' terminal side of each sequence. Among 19 clones, LS3 alone showed the high homology to zebrafish nucleosome assembly protein 1 (NAP1) and the rest of 18 was not yet identified. In situ hybridization confirmed that faint expression of NAP1 mRNA was observed in cells along the ridge of the scale without lateral line. On the other hand, in small cells not along the ridge of the scale with lateral line, very intense hybridization was found as expected. They were abundant at the surrounding area of the lateral line tube. NAP1 plays an enhancing role on gene expression by promoting nucleosome assembly. Thus, cell viability in the scale with lateral line seemed to be higher than that in the scale without lateral line. The scale can be a good model to monitor the effects of environmental pollutants because simultaneous observation against cells with different cell viability is available through comparison between cells in the scale with or without lateral line.

The relationship between post-prandial plasma glucose and post-challenge plasma glucose in Japanese population.

The relationship between post-prandial plasma glucose (PPG) and post-challenge plasma glucose (PCG) within individuals was investigated in Japanese population. The oral glucose tolerance test (OGTT) and measurements of PPG 2h after ingestion of a standardized rice-based meal (PPG2h), were performed in 4471 middle-aged Japanese subjects (2774 men and 1697 women, 50.7+/-8.5 years). There was a loose correlation between PPG2h and PCG2h (r=0.327, p<0.001). The diabetes group (n=170) showed the highest PPG2h, followed by the IGT group (n=786) and the NGT group (n=3414) (p<0.05). At the cutoff point of 140 mg/dl (7.8 mmol/l) for PPG2h, specificities were 94.9% for IGT plus diabetes and 92.9% for diabetes, but sensitivities were as low as 23.2% for IGT plus diabetes and 44.7% for diabetes. The correlation of PPG2h with PCG2h was stronger in the obese group (BMI>or=25 kg/m2) than in the lean group (BMI<20 kg/m2). We conclude that the correlation between PPG2h and PCG2h was significant but not very tight. In evaluating PPG2h, if the cutoff point of 140 mg/dl (7.8 mmol/l) for PCG2h is extrapolated, the majority of subjects with dysglycemia could be overlooked.

Arachidonic acid promotes glutamate-induced cell death associated with necrosis by 12- lipoxygenase activation in glioma cells.

Glutamate induced glutathione (GSH) depletion in C6 rat glioma cells, which resulted in cell death. This cell death seemed to be apoptosis through accumulation of reactive oxygen species (ROS) or hydroperoxides representing cytochrome c release from mitochondria and internucleosomal DNA fragmentation. A significant increase of 12-lipoxygenase enzyme activity was observed in the presence of arachidonic acid (AA) under GSH depletion induced by glutamate. AA promoted the glutamate-induced cell death, which reduced caspase-3 activity and diminished internucleosomal DNA fragmentation. Furthermore, AA reduced intracellular NAD, ATP and membrane potentials, which indicated dysfunction of the mitochondrial membrane. Protease inhibitors such as N-alpha-tosyl-L-phenylalanine chloromethyl ketone (TPCK) and 3, 4-dichloroisocumarin (DCI) but no Ac-DEVD, a caspase inhibitor, suppressed the glutamate-induced cell death. AA reduced the inhibitory effect of TPCK and DCI on the glutamate-induced cell death. These results suggest that AA promotes cell death by inducing necrosis from caspase-3-independent apoptosis. This might occur through lipid peroxidation initiated by ROS or lipid hydroperoxides generated during GSH depletion in C6 cells.

Effects of chlorpromazine on plasma membrane permeability and fluidity in the rat brain: a dynamic positron autoradiography and fluorescence polarization study.

Antipsychotic drugs have been widely used in psychiatry for the treatment of various mental disorders, but the underlying biochemical mechanisms of their actions still remain unclear. Although phenothiazine antipsychotic drugs have been reported to directly interact with the peripheral plasma membrane, it is not known whether these drugs actually affect plasma membrane integrity in the central nervous system. To clarify these issues, we investigated the effect of chlorpromazine (CPZ), a typical phenothiazine antipsychotic drug, on plasma membrane permeability in fresh rat brain slices using a dynamic positron autoradiography technique and [(18)F]2-fluoro-2-deoxy-D-glucose ([(18)F]FDG) as a tracer. Treatment with CPZ (> or =100 microM) resulted in the leakage of [(18)F]FDG-6-phosphate, but not [(18)F]FDG, suggesting that the [(18)F]FDG-6-phosphate efflux was not mediated by glucose transporters, but rather by plasma membrane permeabilization. The leakage of [(18)F]FDG-6-phosphate was followed by slower leakage of cytoplasmic lactate dehydrogenase, suggesting that CPZ could initially induce small membrane holes that enlarged with time. Furthermore, the addition of CPZ (> or =100 microM) caused a decrease in 1,6-diphenyl-1,3,5-hexatriene fluorescence anisotropy, which implies an increase in membrane fluidity. CPZ loading dose-dependently increased both membrane permeability and membrane fluidity, which suggested the involvement of a perturbation of membrane order in the mechanisms of membrane destabilization induced by antipsychotic drugs.