Fluoxetine Up-Regulates Bcl-xL Expression in Rat C6 Glioma Cells.

OBJECTIVE: To analyze both differentially expressed genes and the Bcl-xL protein expression after acute and chronic treatment with fluoxetine in rat C6 glioma cells. METHODS: C6 glioma cells were cultured for 24 h or 72 h after treatment with 10 µM fluoxetine, and gene expression patterns were observed using microarray and qRT-PCR. Then, cells were cultured for 6 h, 24 h, 72 h or 96 h after treatment with 10 µM fluoxetine, and the expression of Bcl-xL protein was measured using western blot. RESULTS: As determined by microarray, treatment with fluoxetine for 24 h up-regulated 33 genes (including Bcl-xL and NCAM140) and down-regulated 7 genes (including cyclin G-associated kinase). Treatment with fluoxetine for 72 h up-regulated 53 genes (including Gsα and Bcl-xL) and down-regulated 77 genes (including Gαi2 and annexin V). Based on the qRT-PCR results, there was an increase in Gsα mRNA and a decrease in Gαi2 mRNA at 72 h in fluoxetine-treated cells as compared to control, a result that was consistent with microarray. We also observed an increase in Bcl-xL mRNA (both at 24 h and at 72 h) in fluoxetine-treated cells as compared to control, demonstrating a tendency to increase gradually. Bcl-xL protein expression increased as the duration of fluoxetine treatment increased. CONCLUSION: These results suggest that chronic treatment with fluoxetine not only initiates the cAMP pathway through inducing Gsα expression but also induces Bcl-xL expression, thus inhibiting apoptosis.

The Effects of Venlafaxine and Dexamethasone on the Expression of HSP70 in Rat C6 Glioma Cells.

OBJECTIVE: The present study aimed to determine the intracellular action of the antidepressant, venlafaxine, in C6 glioma cells using heat shock protein 70 (HSP70) immunocytochemistry and HSP70 Western blots; HSP70 is known to be associated with stress and depression. METHODS: The extent of HSP70 expression was measured after rat C6 glioma cells were treated with 1) dexamethasone only, 2) venlafaxine only, 3) simultaneous venlafaxine and dexamethasone, or 4) dexamethasone after venlafaxine pretreatment. Dexamethasone (10 microM, 6 hours) did not affect the level of HSP70 expression relative to control. RESULTS: Short-term (1 hour) venlafaxine treatment significantly increased the level of HSP 70 expression. Simultaneous long-term (72 hours) venlafaxine and dexamethasone treatment significantly reduced the level of HSP70 expression. Dexamethasone treatment administered following long-term (24 and 72 hours) pretreatment with venlafaxine also significantly reduced the level of HSP70 expression. CONCLUSION: Short-term treatment with venlafaxine increases the expression of HSP70, but prolonged treatment with dexamethasone suppresses the venlafaxine-induced expression of HSP70. These findings suggest that HSP70 and dexamethasone play a significant role in the pathophysiology of depression.

Increased transforming growth factor-beta1 in alcohol dependence.

Ethanol and its metabolite acetaldehyde increase transforming growth factor beta1 (TGF-beta1) expression in animal studies. TGF-beta1 is related with the hepatic stellate cell (the key element of hepatic fibrogenesis) and the radial glia (the key element of neuronal migration). Blood samples were collected from 41 patients with alcohol dependence, TGF-beta1 levels measured by ELISA were compared with 41 normal subjects. Plasma TGF-beta1 levels in the patients with alcohol dependence (1,653.11+/-532.45 pg/mL) were significantly higher than those of healthy subjects (669.87+/-366.53 pg/mL) (P=0.000). Patients with or without liver pathology showed no difference in TGF-beta1 (P=0.36). Increased TGF-beta1 may mediate deleterious effect of alcohol such as hepatic fibrosis and suppressed neuronal developments in alcohol dependence patients.

Relation between plasma brain-derived neurotrophic factor and nerve growth factor in the male patients with alcohol dependence.

Brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) are thought to be related to neuroprotection in cell culture and animal studies. Our aim was to verify the changes in human plasma BDNF and NGF concentrations induced by chronic alcohol use. Forty-one male patients with alcohol dependence were sampled the next morning of admission and compared with 41 healthy male subjects. Plasma BDNF and NGF were assayed using an enzyme-linked immunosorbent assay (ELISA). Mean plasma BDNF level was significantly higher in the patients with alcohol dependence (3502.21+/-1726.9 pg/mL) compared with the healthy subjects (861.75+/-478.9 pg/mL) (P=.000). Mean plasma NGF level was also significantly higher in patients with alcohol dependence (137.64+/-32.7 pg/mL) than in healthy subjects (112.61+/-90.2 pg/mL) (P=.012). Plasma BDNF and NGF levels showed significant negative correlation in alcohol dependence group (r=-0.388, P=.012). Increased plasma BDNF and NGF with negative correlation in alcohol-dependent patients may have some role in the regeneration of damage done by chronic alcohol use.

Effects of haloperidol and risperidone on the expression of heat shock protein 70 in MK-801-treated rat C6 glioma cells.

Non-competitive N-methyl-D-aspartate (NMDA) receptor antagonists such as dizocilpine (MK-801) produce schizophrenia-like psychosis in humans and induce the expression of heat shock protein 70 (HSP70) in rats. The present study examines the effects of antipsychotic drugs, haloperidol and risperidone, on the expression of HSP70 produced by MK-801 in rat C6 glioma cells. After pretreating with haloperidol and risperidone for 1 h, 6 h, 24 h and 72 h, respectively, C6 glioma cells were cultivated again in MK-801 for 6 h, and then, the extent of HSP70 expression was measured by immunoblotting using anti-HSP70 monoclonal antibody. The expression of HSP70 induced by MK-801 significantly decreased as the duration of haloperidol pretreatment was extended (p=0.002). Risperidone also increasingly attenuated the expression of HSP70 produced by MK-801 as the duration of pretreatment grew longer (p=0.003). The present findings show that haloperidol and risperidone decrease the HSP70 expression in MK-801-treated rat C6 glioma cells. These results suggest that HSP70 and NMDA receptors may play a significant role in the pathophysiology of schizophrenia.

Social-cognitive predictors of dietary behaviors in South Korean men and women.

BACKGROUND: Eating a diet that is high in vitamins and low in fat is considered to be governed by social-cognitive factors, such as intentions, planning, self-efficacy, and outcome expectancies. PURPOSE: A longitudinal field study was designed to examine the interrelationships of these factors with dietary behaviors. METHOD: In 697 South Korean men and women, objective health-risk status was assessed at Time 1 (cholesterol, blood pressure, and body mass index) in conjunction with self-efficacy, outcome expectancies, and intentions. At Time 2, six months later, coping self-efficacy, planning, and dietary behaviors were measured. A two-group structural equation model for men and women was specified to determine the relations of distal and proximal predictors of a healthy diet. RESULTS: Self-efficacy was of equal predictive power in men and women, whereas intentions and planning were relevant only in women. Objective risk status was associated with intentions in women but not in men. CONCLUSIONS: Results confirm the predictive power of the Health Action Process Approach and point to the role of gender in the self-regulation of dietary behaviors.

Cue-exposure therapy to decrease alcohol craving in virtual environment.

During abstinence from alcohol, craving is elicited by the cues and contexts previously associated with alcohol, which contribute to relapse. To prevent the craving and relapse experienced by alcoholics, cue-exposure therapy (CET) has been used to extinguish the association between alcohol and alcohol-related cues and contexts. This study applied CET, using a virtual reality (VR) system, to eight members of an Alcoholics Anonymous group for eight sessions. Cues and contexts most likely to elicit an urge to drink were selected through a preliminary survey in order to compose VR-CET scenarios: a glass, a bottle, food, and a bar were judged to be the most tempting for people in alcohol dependence and abstinence. Using these cues and contexts, a Japanese-style pub and a western bar were created. Each session was administered for 30 minutes by a psychiatrist and included an introduction, immersion, VR navigation, interviews about feelings, and self-report questionnaires about cravings. The eight sessions consisted of initial and closing sessions and person-, object-, and situation-focused sessions. As a result, a reduction in cue-elicited craving after VR-CET was reported. A mean score of 15.75 (SD = 10.91) on the Alcohol Urge Questionnaire in the first session decreased to 11.50 (SD = 5.76) in the final session. This study suggests that using virtual reality can enhance the effectiveness of CET.

Detrended fluctuation analysis of resting EEG in depressed outpatients and healthy controls.

OBJECTIVE: Recent findings have demonstrated that the EEG possesses long-range temporal (auto-) correlations (LRTC) in the dynamics of broad band oscillations. The analysis of LRTC provides a quantitative index of statistical dependencies in oscillations on different time scales. We analyzed LRTC in resting EEG signals in depressed outpatients and healthy controls. METHODS: The participants in this study were 11 non-depressed, age-matched controls, and 11 unmedicated unipolar depressed patients. EEG data were obtained from each participant during 5-min resting baseline periods with eyes closed and then analyzed with detrended fluctuation analysis (DFA), a scaling analysis method that quantifies a simple parameter to represent the correlation properties of a time series. The scaling exponent, the result of DFA, provides a quantitative measure of LRTC from the EEG. RESULTS: The present study demonstrates that all the scaling exponents in depressed patients and healthy controls were greater than 0.5 and less than 1.0, regardless of condition. Furthermore, the scaling exponents of depressed patients have relatively higher values in whole brain regions compared to healthy controls, with significant differences at F3, C3, T3, T4 and O1 channels (p<0.05). Finally, a significant linear correlation was observed between the severity of depression and the scaling exponent over most of the channels, except O2. CONCLUSIONS: These results suggest that the brain affected by a major depressive disorder shows slower decay of the LRTC, and that the persistence of the LRTC of EEG in depressed patients was associated with the severity of depression over most of the cortical areas. SIGNIFICANCE: The DFA method may broaden our understanding of the psychophysiological basis of depression.

Changes of brain potentials in response to smoking-induced stimuli in smokers.

Changes in P300 amplitude were used as an indicator of reactivity to smoking-related stimuli in smokers. The amplitude of P300--a component of event-related brain potentials (ERPs) elicited by 10 smoking-related (craving), 10 antismoking (aversive) and 10 neutral stimuli-- was recorded in smokers (n=10) and nonsmokers (n=10). Electroencephalography (EEG) data were obtained by the Laxtha EEG-monitoring device in the EEG recording room, and were recorded at F3, F4, C3, and C4. Three-way repeated-measures analysis of variance (ANOVA) was computed on the P300 amplitudes. The factors were group (smokers, nonsmokers), stimulus (craving, aversive, neutral), and electrode location (F3, F4, C3, and C4). The main effects of stimulus were significant, but the group effects did not show significant interactions with other factors. An interesting observation was the similarity between P300 waveforms for craving and aversive stimuli in smokers. These findings could indicate that the antismoking-related response is similar to the smoking-related one.

Fractal analysis of EEG in hypnosis and its relationship with hypnotizability.

Fractal analysis was applied to study the trends of EEG signals in the hypnotic condition. The subjects were 19 psychiatric outpatients. Hypnotizability was measured with the Hypnotic Induction Profile (HIP). Fifty-four sets of EEG data were analyzed by detrended fluctuation analysis (DFA), a well-established fractal analysis technique. The scaling exponents, which are the results of fractal analysis, are reduced toward white noise during the hypnotic condition, which differentiates the hypnotic condition from the waking condition. Further, the decrease in the scaling exponents during hypnosis was solely associated with the eye-roll sign within specific cortical areas (F3, C4, and O1/2) closely related to eye movements and attention. In conclusion, the present study has found that the application of the fractal analysis technique can demonstrate the electrophysiological correlations with hypnotic influence on cerebral activity.

Naltrexone influences protein kinase Ce and integrin alpha7 activity in SH-SY5Y neuroblastoma cells.

Alcohol influences the neuroadaptation of brain cells where receptors and enzymes like protein kinase C (PKC) exist. Naltrexone acts on opioid receptors. However, other mechanisms of action remain unknown. We prepared SH-SY5Y neuroblastoma cells, and fed them with 150 mM ethanol for 72 h followed by treatment with naltrexone for 24 h. We performed microarray analysis and reverse transcriptase-polymerase chain reaction. Our results showed that PKCepsilon increased 1.90 times and showed an overall decreasing pattern as time increased. Phosphorylated ERK also increased 2.0 times according to the change of PKCepsilon. Integrin alpha7 increased 2.32 times and showed an increasing pattern as time increased. In conclusion, naltrexone influences PKCepsilon neuronal signaling system and endothelial adhesion molecule integrin alpha7 in addition to the well-known opioid system.

Cardiomyopathy associated with clozapine.

Clozapine is an atypical antipsychotic agent that is more effective than the standard neuroleptics currently used for treating refractory schizophrenia. In addition, clozapine is a drug with few extrapyramidal side effects. However, clozapine is also associated with potentially serious adverse effects, such as cardiac complications as well as agranulocytosis. Clozapine-related cardiomyopathy has not been previously reported in East Asia. This report describes a 31-year-old Korean male patient with schizophrenia who developed dilated cardiomyopathy on treatment with clozapine. The removal of clozapine caused subsequent physical improvement. However, the readministration of clozapine for managing relapse of psychosis caused a recurrence of dilated cardiomyopathy in this patient. Therefore, this is the 1st report showing that the 2nd trial of clozapine caused recurrence of cardiomyopathy associated with clozapine. Thus, this report adds important support for a causal relation between clozapine and cardiac complications. In conclusion, this report attempts to raise awareness of clozapine-related cardiomyopathy.

Genetic polymorphisms of alcohol and aldehyde dehydrogenase, dopamine and serotonin transporters in familial and non-familial alcoholism.

One hundred and eleven male patients with alcohol dependence and 123 nonalcoholic healthy men were tested for the genetic polymorphisms of alcohol dehydrogenase 2 (ADH2), aldehyde dehydrogenase 2 (ALDH2), serotonin transporter (5-HTT) and dopamine transporter (DAT1). There were significant differences in genotype frequencies of ADH2 C992G and A13543G SNPs between alcoholic patients with family history of alcohol dependence (familial) and alcoholic patients without family history (non-familial). Genotype and allele frequencies of ALDH2 G1951A SNP in familial or non-familial alcoholic patients differ from normal controls. Neither 5-HTTLPR L/S nor DAT1 G2319A SNP genotypes nor alleles discriminated alcoholic patients from normal controls. These findings suggest that the genetic characteristics of alcohol metabolism in non-familial alcoholics fall between non-alcoholism and familial alcoholics.

Scanning of genetic effects of alcohol metabolism gene (ADH1B and ADH1C) polymorphisms on the risk of alcoholism.

Alcoholism is a multifactorial and polygenic disorder involving complex gene-to-gene and gene-to-environment interactions. Alcohol metabolism is one of the biological determinants that could significantly be influenced by genetic polymorphisms in alcohol-metabolism genes. These genetic polymorphisms are believed to influence drinking behavior and development of alcoholism. Direct DNA sequencing of whole ADH1B and ADH1C genes revealed 36 sequence variants, including six nonsynonymous and 14 novel polymorphisms. Seventeen polymorphisms among them were selected for genotyping in a larger study (n = 352) based on linkage disequilibria (LDs) among SNPs, locations, and frequencies. Hardy-Weinberg equilibrium (HWE) analyses of polymorphisms revealed severe deviations only in alcoholics, which strongly suggest that a selection bias (or pressure) may be involved. The analyses of genotype distribution in alcoholics (n = 106) and normal controls (n = 246) showed dramatic associations with the risk of alcoholism. Fourteen polymorphisms in ADH1C and ADH1B showed a series of different strengths of association and magnitudes of risk. Based on referent and subgroup analysis, it was strongly suggested that the genetic effects come from the ADH1B*47Arg/*47Arg genotype, and that the positive signals from other sites are just tracking the genetic effect of ADH1B His47Arg. In this article we present summaries of previous studies and of the present study, to give an overview of the worldwide effects of ADH1B His47Arg on the risk of alcoholism. The information derived from this study could be valuable for understanding the genetic factors involved in the risk of alcoholism and facilitate further investigation in other ethnic groups.

Fluoxetine-induced up-regulation of 14-3-3zeta and tryptophan hydroxylase levels in RBL-2H3 cells.

The primary mechanisms of antidepressants are based on the monoamine depletion hypothesis. However, we do not yet know the full cascade of mechanisms responsible for the therapeutic effect of antidepressants. To identify the genes involved in the therapeutic mechanism of the selective serotonin reuptake inhibitor, fluoxetine, we used a cDNA microarray analysis with RBL-2H3 cells. We observed the transcriptional changes of several tens of genes containing the 14-3-3zeta gene in the fluoxetine-treated RBL-2H3 cells. Real-time RT-PCR and Western blotting confirmed changes in the expression of the gene and protein. The increase of 14-3-3zeta mRNA was observed at 72 h in the fluoxetine-treated RBL-2H3 cells. The increase of 14-3-3zeta protein was observed at 48 and 72 h. In this study, the expressions of the 14-3-3zeta gene and the protein were up-regulated at 72 h. In addition, the increase of TPH mRNA was observed at 12, 24 and 72 h in the fluoxetine-treated RBL-2H3 cells. We conclude that fluoxetine induces increases of 14-3-3zeta mRNA, 14-3-3zeta protein and TPH mRNA at 72 h in the RBL-2H3 cells. This suggests that the 14-3-3zeta and TPH genes may play a role in the molecular mechanism of fluoxetine. To date, no cases of 14-3-3zeta alterations by antidepressants and specifically by fluoxetine have been reported.

Dexamethasone inhibits proliferation of adult hippocampal neurogenesis in vivo and in vitro.

Activation of glucocorticoid receptor (GR) induces a reduction of adult hippocampal neurogenesis found in dentate gyrus (DG). However, the nature of specific effects by glucocorticoid in hippocampal neurogenesis is not known. In this report, we show differential effects of dexamethasone (DEX), a glucocorticoid receptor agonist, on proliferation and functional differentiation of adult hippocampal progenitor cells in DG. Two-month-old adult rats received daily injections of DEX for 9 days and were sacrificed 12 h and 28 days after the ninth injection. Proliferation assays showed that DEX inhibited proliferation of neural progenitor cells and the inhibitory effects of DEX was not detected 28 days after recovery. Functional differentiation studies using B-cell lymphoma protein-2 (Bcl-2), brain-derived neurotrophic factor (BDNF), p-ERK, and neuronal nuclear protein (NeuN) antibodies revealed that the expressions of Bcl-2 and BDNF were not significantly different between control and DEX-treated rats. In contrast, however, the activation of extracellular signal-regulated kinase (ERK) was downregulated 12 h, but not 28 days, after the DEX treatment. When adult hippocampal progenitor cell cultures were treated with subchronic DEX, proliferation of the progenitor cells was suppressed. Taken these in vitro and in vivo results together, it is concluded that glucocorticoid receptor activation blocks only proliferation, but not differentiation, in hippocampal neurogenesis.

Phosphorylation of ERK and CREB in cultured hippocampal neurons after haloperidol and risperidone administration.

The purpose of the present paper was to determine whether the brief exposure of neurons to antipsychotic drugs is associated with the activation of extracellular signal-regulated kinases (ERK) and cyclic adenosine 3',5'-monophosphate (cAMP) response element (CRE) binding protein (CREB). The activation of ERK-1/2 and CREB can be monitored by immunoblotting with antibodies that specifically recognize p-ERK-1/2 (phosphorylated on Thr-202 and Tyr-204) and p-CREB (phosphorylated on Ser-133). In hippocampal neuron cultures at 25 days in vitro (DIV), the levels of ERK and CREB phosphorylation significantly increased after treatment with haloperidol (50 nmol/L) and risperidone (50 nmol/L), except when risperidone was administered at the p-CREB level. However, risperidone also increased the p-CREB level at an insignificant rate in the same direction. At 10 DIV, none of the antipsychotic drugs significantly increased the level of ERK and CREB phosphorylation. The difference between levels of ERK and CREB phosphorylation in response to haloperidol and risperidone at 25 DIV was also observed. Risperidone significantly increased the level of ERK-1/2 phosphorylation, but not the level of CREB phosphorylation. Haloperidol, in contrast, had a different effect. These data indicate that neuronal maturation affects the phosphorylation of ERK and CREB in response to antipsychotic drugs. Furthermore, these results demonstrate that different antipsychotic drugs could lead to different profiles of ERK and CREB phosphorylation in neurons.

Korean version of the diagnostic interview for genetic studies: Validity and reliability.

The Diagnostic Interview for Genetic Studies (DIGS), developed in 1994 by the National Institute of Mental Health (NIMH), was translated into Korean and tested for reliability and diagnostic validity. Concurrent validity was tested using the Structured Clinical Interview for DSM-IV (SCID) and clinical diagnoses in 53 patients, most of whom had either schizophrenia or bipolar disorder. Inter-rater reliability was tested in 24 patients. Test-retest reliability was also tested in 17 patients. Overall and specific diagnostic validity for the Korean version of DIGS (DIGS-K) was excellent for most diagnoses. Inter-rater and test-retest reliability for overall and specific diagnoses also ranged from fair to excellent. For schizoaffective disorder, the test-retest reliability of DIGS-K was in a fair range, although the level was lower than that of other diagnoses. However, its diagnostic validity and inter-rater reliability was below fair range. In conclusion, DIGS-K appears to be a reliable interview for major psychiatric disorders.

Experimental application of virtual reality for nicotine craving through cue exposure.

Research has shown that many smokers experience an increase in the desire to smoke when exposed to smoking-related cues. Cue exposure treatment (CET) refers to the manualized, repeated exposure to smoking-related cues, aimed at the reducing cue reactivity by extinction. In this study, we constructed a virtual reality system for evoking a desire of nicotine, which was based on the results of a Questionnaire of Nicotine-craving. And we investigated the effectiveness of the virtual reality system as compared to classical device (pictures). As a result, we reached the conclusion that virtual reality elicits more craving symptoms than the classical devices.

Dextromethorphan alters gene expression in rat brain hippocampus and cortex.

Dextromethorphan is a widely used anti-tussive drug with non-competitive antagonistic effects on excitatory amino acid receptors of the N-methyl-D-aspartate (NMDA) type. This study examined the effect of daily dextromethorphan administration on gene expression in rat brain hippocampus and cortex regions using Rat 5K cDNA microarrays. Triplicate microarray assays were performed at each time point (1, 3 and 10 days), and results were confirmed using semi-quantitative RT-PCR on a subset of differentially expressed cDNA. The microarray analysis proved able to detect changes in gene expression following dextromethorphan injection. Moreover, these changes were mostly mediated by an NMDA receptor. The hippocampus region showed more alterations in gene expression than cerebral cortex following dextromethorphan treatment. The expression of many glutamate-induced apoptosis-related genes, and NO-dependent apoptosis-associated genes, was down-regulated. Expression of anti-apoptotic genes, such as nucleophosmin/B23, Rab2, MAP kinase kinase and CREB binding protein, was up-regulated by dextromethorphan. Angiogenesis is likely to be inhibited in our system due to observed down-regulation of VEGF-associated genes. Expression of some SNARE genes was up-regulated in rat brain hippocampus and cortex regions after dextromethorphan injection.