An Unusual Case of Drunk Driving in Japan: Alcohol-Related Sleepwalking.

Alcohol has been identified as a potential precipitating factor for parasomnia, particularly sleepwalking (SW). We report an unusual case of a Japanese drunk driver who may have experienced alcohol-related SW, based on the statements of the suspect, pharmacokinetic analyses of the suspect's breath alcohol concentration, testimonies of witnesses, driving recorder data, and medical records. The existence of sleep-related criminal acts performed while a suspect experiences memory loss under the influence of alcohol has not been sufficiently recognized, and awareness of such acts should be raised among the police, public prosecutors, and the general public in Japan.

Alcohol Consumption in Combination with an Atherogenic Diet Increased Indices of Atherosclerosis in Apolipoprotein E/Low-Density Lipoprotein Receptor Double-Knockout Mice.

BACKGROUND: Alcohol abuse and adherence to atherogenic diet (AD; a low-carbohydrate-high-protein diet) have been positively associated with cardiovascular disease. In addition, it has been demonstrated clinically that dietary intake is increased on days when alcohol is consumed. Here, the additive effects of ethanol (EtOH) and AD on atherosclerosis, a major underlying cause of cardiovascular disease, were investigated in apolipoprotein E/low-density lipoprotein receptor double-knockout (KO) mice. The mechanisms, especially aortic oxidative stress damage, were highlighted. METHODS: Twelve-week-old male KO mice on AD with or without EtOH treatment were bred for 4 months. Age-matched male C57BL/6J mice on a standard chow diet without EtOH treatment served as controls. Analyses were conducted using ultrasound biomicroscopy, histopathological and fluorescence immunohistochemical examinations, Western blots, and polymerase chain reaction. RESULTS: KO mice on AD with EtOH treatment showed increases in aortic maximum intima media thickness, hypoechoic plaque formation, and mean Oil-Red-O content. These results were associated with enhanced ratio of aortic 8-hydroxy-2'-deoxyguanosine (8-OHdG)-immunopositive area to the metallothionein (MT) immunopositive area and suppression of AD-induced up-regulated aortic Mt1, Mt2, and upstream stimulatory factor 1 mRNA expressions. Moreover, 8-OHdG was expressed in the nuclei of CD31- and alpha smooth muscle actin-immunopositive cells, and the up-regulated mRNA expressions of aortic nitric oxide synthase 3 and platelet-derived growth factors were only observed in the KO mice on AD with EtOH treatment. CONCLUSIONS: Alcohol abuse and adherence to AD may promote the shift of aortic oxidative stress and antioxidative stress balance toward oxidative stress predominance and reduced antioxidative stress, which may be partly due to the decrease in MT at the cell biological level and down-regulation of Mt at the gene level, which in turn could play a role in the up-regulation of endothelial dysfunction-related and vascular smooth muscle cell proliferation-related gene expression and the progression of atherosclerosis in mice with hyperlipidemia.

Effect of Alcohol Sensitivity in Healthy Young Adults on Breath Pharmacokinetics of Acetaldehyde After Mouth Washing with Alcohol.

BACKGROUND: Acetaldehyde is causally related to head and neck cancer. Individuals with aldehyde dehydrogenase 2 deficiency experience alcohol sensitivity and are referred to as "flushers" because of their skin-flushing response to high blood acetaldehyde levels after alcohol consumption. Acetaldehyde is produced in the oral cavity after local alcohol exposure without alcohol ingestion. However, the relationship between the oral acetaldehyde level after local alcohol exposure and alcohol sensitivity is unclear. Herein, sampling the exhaled breath, we evaluated the effect of alcohol sensitivity on the pharmacokinetics of ethanol (EtOH) and acetaldehyde in breath after mouth washing with alcohol. METHODS: Twenty-eight healthy young adults were divided into flusher and nonflusher groups based on an EtOH patch test. The subjects washed their mouths for 30 seconds with 40 ml of 5% v/v alcohol, and their breath samples were collected 12 times over 20 minutes after mouth washing and rinsing with water. EtOH and acetaldehyde concentrations in all breath samples were measured using sensor gas chromatography. RESULTS: Breath EtOH concentrations exponentially decreased in both groups after mouth washing with alcohol. Breath acetaldehyde concentrations showed an immediate increase, followed by an almost exponential decrease in both groups, but concentrations in the flusher group remained higher than those in the nonflusher group throughout the 20-minute measurement period. This was reflected in a peak concentration (Cmax ) of 808 ± 70 parts-per-billion (ppb) versus 1,715 ± 223 ppb, respectively (p = 0.001), and area under the curve values of 3,528 ± 1,399 ppb minutes versus 8,637 ± 1,293 ppb minutes, respectively (p = 0.002). CONCLUSIONS: This study revealed high concentrations of acetaldehyde in breath after local alcohol exposure in the oral cavity among flushers even without alcohol ingestion. This contributes to an increased risk among flushers of mutagenic DNA lesions in the mucosa of the upper digestive tract and cancer.

Pharmacokinetic analyses using absorption kinetics in low-alcohol dose cases of drunken driving.

In Japan, low-alcohol dose cases of drunken driving, where drivers drink just before getting behind the wheel, are increasing for expert witnesses since the penalties for drunken driving have become stricter. Widmark's equation has generally been used for the pharmacokinetic analysis of blood alcohol concentration, which encompasses the one-compartment model with zero-order elimination kinetics but ignores absorption kinetics. We therefore propose that the formula might not be applicable to the analysis of low-alcohol dose cases of drunken driving because the issue is focused on the absorption phase. In this paper, we present two representative low-alcohol dose cases, which were analyzed using the one-compartment model with first-order absorption and zero-order elimination kinetics. This formula is thought to be more suitable and useful for medicolegal practice than Widmark's formula.

Acute restraint stress provokes sudden cardiac death in normotensive rats and enhances susceptibility to arrhythmogenic effects of adrenaline in spontaneously hypertensive rats.

BACKGROUND: A high incidence of cardiovascular events and sudden cardiac death (SCD) has been reported following unexpected acute psychosocial stress. The possible pathways by which acute restraint stress (ARS), a kind of acute psychosocial stress, leads to SCD were determined. METHODS: Using 16-week-old male normotensive Wistar Kyoto rats (WKY, n=24) as controls and spontaneously hypertensive rats (SHR, n=24) as the hypertensive subjects with left ventricular hypertrophy (LVH), we assessed ARS-related incidence of SCD, cardiac and myocardial autonomic nervous system dysfunction, gap junction connexin-43 (Cx43) channel remodeling, and ventricular repolarization abnormality, based on electrocardiography, an adrenaline test, heart rate variability (HRV), and reverse transcriptase polymerase chain reaction analyses. Rats with ARS were introduced into restrainers that allowed head, limb, and tail movement. RESULTS: In normotensive hearts without LVH, ARS induced a higher incidence of SCD attributed to lethal bradycardia, increased cardiac and myocardial sympathetic activation, and gap junction Cx43 channel remodeling, as evidenced by the increases in the ratio of low-frequency and high-frequency powers in HRV, the ratio of myocardial neuropeptide Y (NPY) and acetylcholinesterase (AChE) mRNA expressions, and the up-regulation of LV Cx43 mRNA expression; in hypertensive hearts with LVH, ARS enhanced susceptibility to the malignant arrhythmogenic effects of the adrenaline test (a kind of sympathetic stimuli) accompanied by abnormal ventricular repolarization, as evidenced by increased incidence of ventricular tachycardia and/or ventricular fibrillation during the adrenaline test and prolonged QTc immediately after ARS. CONCLUSIONS: ARS may trigger cardiac and myocardial sympathetic predominance, and then induce gap junction Cx43 channel remodeling, finally leading to lethal bradycardia in normotensive WKY. ARS-induced abnormal ventricular repolarization may be responsible for ARS-enhanced susceptibility to sympathetic stimulation in SHR with LVH. Expressions of myocardial NPY, AChE, and Cx43 genes, HRV, QTc and LVH measures showed diagnostic and prognostic potential for predicting ARS-induced SCD.

An autopsy case of acetyl fentanyl intoxication caused by insufflation of 'designer drugs'.

We present a fatal case of intoxication due to insufflation of acetyl fentanyl. His blood concentration of acetyl fentanyl was 270ng/mL, and the manner of death was classified as an accident. This is the first report of an autopsy case of acetyl fentanyl delivered by insufflation, rather than intravenous administration. He had been snoring loudly for at least 12h prior to death, and transport to a hospital during this time and treatment with naloxone may have saved his life. In this sense, it can be said that his death was preventable. This case reemphasizes the risk of death associated with drug overdose and the narrow range of acetyl fentanyl between the effective dose (ED50) and lethal dose (LD50). The case should also raise awareness among medical professionals of the effectiveness of naloxone and the need to establish a comprehensive system for toxicological analysis while keeping the possibility of use of 'designer drugs' in mind.

Angiotensinase C mRNA and Protein Downregulations Are Involved in Ethanol-Deteriorated Left Ventricular Systolic Dysfunction in Spontaneously Hypertensive Rats.

The influences of angiotensinase C on ethanol-induced left ventricular (LV) systolic function were assessed in spontaneously hypertensive rats (SHRs). SHRs were fed by a liquid diet with or without ethanol for 49 days. The normotensive Wistar Kyoto rats (WKY) were fed by the liquid diet without ethanol and used as control. We evaluated LV systolic function, angiotensinase C mRNA and protein expressions, activation of the renin-angiotensin system (RAS), and the gene expressions of LV collagen (Col) III a1 and matrix metalloproteinases- (MMP-) 9. Compared to the WKY, LV systolic dysfunction (expressed by decreased fractional shortening and ejection fraction) was observed in the SHRs before ethanol treatment and further deteriorated by ethanol treatment. In the ethanol-treated SHRs, the following were observed: downregulations of angiotensinase C mRNA and protein, increased RAS activity with low collagen production as evidenced by angiotensin II and angiotensin type 1 receptor (AT1R) protein upregulation, AT1aR mRNA downregulation, and an MMP-9 mRNA expression upregulation trend with the downregulation of Col III a1 mRNA expression in LV. We conclude that chronic ethanol regimen is sufficient to promote the enhanced RAS activity-induced decrease in the production of cardiac collagen via downregulated angiotensinase C, leading to the further deterioration of LV systolic dysfunction in SHRs.

[Relationship between alcohol consumption and external causes of death based on the forensic autopsy cases in Yamaguchi].

We analyzed forensic autopsy cases to assess the relationship between alcohol consumption and external causes of death. We divided 605 autopsy cases which had been performed from 2000 to 2011 at our department into Alcohol group (n = 172, 28.4%) and Non-alcohol group (n = 433, 71.6%) according to whether alcohol could be detected in the deceased's blood. The individuals' sex and age, season when the death occurred, cause of death, type of death and circumstances of death were analyzed. Alcohol group had a significantly higher ratio of males and younger ages (both p < 0.05). There was no significant between-group difference regarding the seasons when the deaths occurred. Alcohol group had significantly greater rates of spinal injuries, abdominal injuries, traffic accidents, and accidental drowning. "Bicycling" was revealed as a related factor of the traffic accidents only in Alcohol group. In contrast, "accident on the expressway," "riding a motorcycle," and "a passenger in a vehicle" were related factors only in Non-alcohol group. We concluded that the factors of male gender and middle-to-senior age are responsible for the increased risk of external causes of death after alcohol consumption, and that alcohol consumption is one of the risk factors for accidental death. In Japan, drunk-driving-related accidents have shown a downward trend whereas bicycling-related accidents have shown an upward trend, and similar results were obtained in the present study. The low awareness of drinking-induced pitfalls may be responsible for drinking-related bicycle accidents.

Effects of hypothermia on ex vivo microglial production of pro- and anti-inflammatory cytokines and nitric oxide in hypoxic-ischemic brain-injured mice.

INTRODUCTION: Activated microglia produce neurotoxic factors, including pro-inflammatory cytokines and nitric oxide (NO), in response to neuronal destruction. Therapeutic suppression of microglial release of these factors by various approaches including hypothermia is considered to be neuroprotective after severe brain damage. We examined the effects of hypothermic culture on the production of pro- and anti-inflammatory cytokines and NO in ex vivo microglia that were derived from mice with hypoxic-ischemic (HI) brain injury, through the stimulation of toll-like receptors (TLRs) that play significant roles in the pathological processes underlying a sterile central nervous system injury. MATERIAL AND METHODS: Two-day-old mice underwent the right common carotid artery ligation followed by 6% oxygen for 30 min, and thereafter were placed at 37°C for 24 h, after which microglia were isolated and then cultured with TLR2 and TLR4 agonists at 33°C and 37°C. Cytokine and NO levels in culture supernatants were measured. RESULTS: Compared with 37°C, hypothermia (33°C) reduced the production of tumour necrosis factor-alpha (TNF-α: a pro-inflammatory cytokine) at 6 h and interleukin-10 (IL-10: an anti-inflammatory cytokine) and NO at 48 h. CONCLUSIONS: In TLR-activated microglia that were derived from mice with HI brain injury, hypothermia reduced the production of TNF-α, IL-10, and NO temporally, a clinically relevant finding suggesting that neuroprotection conferred by therapeutic hypothermia is related to attenuation of early-phase and late-phase inflammatory factors as well as that of late-phase anti-inflammatory factor(s) released from microglia.

Carvedilol improves ethanol-induced liver injury via modifying the interaction between oxidative stress and sympathetic hyperactivity in rats.

AIM: Oxidative stress is a major pathway mediating ethanol hepatotoxicity and liver injury. We previously found that carvedilol, which can block the sympathetic nervous system via ß1-, ß2- and α1-adrenoreceptors, modifies ethanol-induced production of lipogenesis- and fibrogenesis-related mediators from hepatic stellate cells (HSC). In the present study, we assessed the effects of carvedilol on ethanol-induced liver injury, hepatic insulin resistance, and the interaction between oxidative stress and sympathetic hyperactivity in rats with alcoholic fatty liver disease (AFLD). METHODS: Male Wistar rats were pair-fed for 49 days and divided into four groups: control and ethanol liquid-diet-fed rats with and without 7-day carvedilol treatment. Rats' sympathetic activity, hepatic oxidative stress, hepatic insulin resistance and liver injury were evaluated based on biochemical analysis, enzyme-linked immunosorbent assay, fluorescence immunohistochemistry, western blot and reverse transcriptase polymerase chain reaction. RESULTS: Forty-nine days of ethanol consumption induced the increases in circulating noradrenaline metabolite (3-methoxy-4-hydroxyphenylglycol), hepatic noradrenaline and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels, the downregulation of hepatic insulin receptor substrate-1 gene expression, and the accumulation of fatty droplets within hepatocytes with the increased hepatic triglyceride and blood alanine aminotransferase levels. All of these changes were modified by carvedilol treatment. 8-OHdG was detected in activated HSC and suppressed by carvedilol treatment based on fluorescence immunohistochemical double-staining analysis. CONCLUSION: Carvedilol may modify the interaction between the oxidative stress and the sympathetic hyperactivity, and then contribute to attenuating the development of AFLD in rats. Additionally, oxidative stress may be responsible for the activation of HSC during the early stage of alcoholic liver disease.

Delayed ethanol elimination and enhanced susceptibility to ethanol-induced hepatosteatosis after liver resection.

AIM: To investigate ethanol-induced hepatic steatosis after liver resection and the mechanisms behind it. METHODS: First, the preliminary examination was performed on 6 sham-operated (Sham) and 30 partial hepatectomy (PH) male Wistar rats (8-wk-old) to evaluate the recovery of the liver weight and liver function after liver resection. PH rats were sacrificed at the indicated time points (4, 8, and 12 h; 1, 3, and 7 d) after PH. Second, the time point for the beginning of the chronic ethanol exposure (1 wk after sham- or PH-operation) was determined based on the results of the preliminary examination. Finally, pair-feeding was performed with a controlled diet or with a 5-g/dL ethanol liquid diet for 28 d in another 35 age-matched male Wistar rats with a one-week recovery after undergoing a sham- (n = 15) or PH-operation (n = 20) to evaluate the ethanol-induced liver injury after liver resection. Hepatic steatosis, liver function, fatty acid synthase (Fas) gene expression level, the expression of lipid metabolism-associated enzyme regulator genes [sterol regulatory element binding protein (Srebp)-1 and peroxisome proliferator-activated receptor (Ppar)-α], the mediators that alter lipid metabolism [plasminogen activator (Pai)-1 gene expression level and tumor necrosis factor (Tnf)-α production], and hepatic class-1 alcohol dehydrogenase (Adh1)-associated ethanol elimination were investigated in the 4 groups based on histological, immunohistochemical, biochemical, Western blotting, reverse transcriptase chain reaction, and blood ethanol concentration analyses. The relevant gene expression levels, liver weight, and liver function were assessed before and 1 wk after surgery to determine the subject's recovery from the liver resection using the rats that had been subjected to the preliminary examination. RESULTS: In the PH rats, ethanol induced marked hepatic steatosis with impaired liver functioning, as evidenced by the accumulation of fatty droplets within the hepatocytes, the higher increases in their hepatic triglyceride and blood alanine aminotransferase and blood aspartate aminotransferase levels after the 28-d pair-feeding period. The Sham-ethanol rats, not the PH-ethanol rats, demonstrated the up-regulation of Srebp-1 and the down-regulation of Ppar-α mRNA expression levels after the 28-d pair-feeding period. The 28-d ethanol administration induced the up-regulation of Pai-1 gene expression level and an overproduction of TNF-α in the Sham and the PH rats; however, the effect was more significant in the PH rats. The PH-ethanol rats (n = 4) showed higher residual blood ethanol concentrations than did the Sham-ethanol rats (n = 6) after a 5-h fast (0.66 ± 0.4 mg/mL vs 0.2 ± 0.1 mg/mL, P < 0.05); these effects manifested without up-regulation of Adh1 gene expression, which was present in the Sham-ethanol group after the 28-d pair-feeding period. One week after the liver resection, the liver weight, function, the gene expression levels of Fas, Srebp-1, Ppar-α, Pai-1 and Tnf-α recovered; however, the Adh1 gene expression did not recover in rats. CONCLUSION: Desensitization to post-hepatectomy ethanol treatment and slow recovery from PH in Adh1 gene expression enhanced the susceptibility to ethanol-induced hepatic steatosis after PH in rats.

[The effects of chronic alcohol exposure on progression of liver injury in spontaneously hypertensive rats; preliminary study].

The association between alcohol intake and blood pressure is well known, and our previous studies indicate that the stimulus of sympathetic nervous system induce the progression of liver injury. In this study, we examined the effects of chronic ethanol treatment on the progression of liver injury using the spontaneously hypertensive rat (SHR). The advantage of using the present strain is to possess sympathetic facilitation without any treatment. Normotensive Wistar-Kyoto rat (WKY) was used as control. 7-week-old male rats were pair-fed with either ethanol- or control-liquid-diet for 49 days and divided into four groups: control liquid-diet-fed WKY and SHR, continuous ethanol liquid diet-fed WKY and SHR. Plasma alanine aminotransferase (ALT) levels, and histological analyses based on Hematoxylin-Eosin (H-E), Oil red O and Sirius red stains of the liver sections were used to assess alcohol-induced liver injury. Chronic ethanol treatment induced the increases in plasma ALT, the accumulation of fatty droplets within hepatocytes and pericellular hepatic fibrosis, particularly in SHR. Between the control group rats of SHR and WKY, SHR showed the increases in accumulation of fatty droplets and pericellular hepatic fibrosis. No significant inflammatory cell infiltration was shown in all groups. These results suggested that chronic ethanol treatment in SHR could induce the more severe liver injuries when compared with WKY. In conclusion, chronic alcohol intake in rats with hypertension could deteriorate the ethanol-induced liver injury via the sympathetic overactivity.

Carvedilol attenuates the progression of alcohol fatty liver disease in rats.

BACKGROUND: Hepatosteatosis is an essential step in liver disease progression. However, the mechanisms that mediate the progression of hepatosteatosis and the optimal inhibitor of them remain largely unclear. The sympathetic nervous system (SNS) is responsible for the lipid metabolism and the accumulation of collagen that occurs in an injured liver. Medicines that inhibit this pathway may be a relevant treatment for the hepatosteatosis, and then reduce the liver injury that progresses through the stage of steatosis to fibrosis. METHODS: Using an ethanol-liquid-diet-fed rat model of alcohol fatty liver disease (AFLD), we studied the effects of carvedilol, which can block the SNS completely via ß1, ß2, and α1 adrenergic receptors, on the sympathetic tone, hepatosteatosis, and fibrosis based on histological, immunohistochemical, Western blot, and reverse transcriptase polymerase chain reaction analyses. RESULTS: Carvedilol inhibited the ethanol-induced whole-body and hepatic sympathetic activities based on the serum 3-methoxy-4-hydroxyphenylglycol level and hepatic tyrosine hydroxylase expression. Carvedilol attenuated the hepatosteatosis, as evidenced by reduced hepatic triglyceride level and the accumulation of fatty droplets within hepatocytes, down-regulated fatty acid synthase and sterol regulatory element binding protein-1, and up-regulated peroxisome proliferator-activated receptor-α. No fibrosis signs were shown in our rat model. Carvedilol inhibited ethanol-induced the thickening of zone 3 vessel walls, reduced the activation of hepatic stellate cells (HSCs), and decreased the induction of collagen, transforming growth factor ß1, and tissue inhibitor of metalloproteinases-1. Tumor necrosis factor α (TNF-α) was expressed on the activated HSCs and inhibited by carvedilol based on the immunohistochemical double staining analysis. CONCLUSIONS: Ethanol metabolism-induced lipogenesis may trigger the SNS-activated HSCs feedback loop, and then induct the activated HSCs and the activated HSCs-derived TNF-α, the mediator of lipogenesis, overproduction. Carvedilol may block this feedback loop via antisympathetic activity and demonstrate its preventive role on the development of hepatosteatosis in rat with AFLD.

Importance of explanation before and after forensic autopsy to the bereaved family: lessons from a questionnaire study.

To investigate how bereaved families felt about the explanation received before and after forensic autopsies, the authors conducted a cross-sectional survey of the bereaved families whose next of kin underwent a forensic autopsy at the two Departments of Forensic Medicine and a few bereaved families of crime victims. Of 403 questionnaires sent, 126 families responded. Among 81.5% of the respondents who received an explanation from policemen before the autopsy, 78.8% felt that the quality of the explanation was poor or improper. In Japan, the law has restricted disclosure of information from a forensic autopsy. Despite legal restrictions, 82% wanted to hear from the person who conducted the autopsy. However, police explained the results of autopsy to 65.2% of respondents. Among the families whose frustration and anger increased after autopsy, 86.4% had not been satisfied with the explanation before the autopsy. Additionally, 57.7% had not been informed on the autopsy findings at the time of the questionnaire when more than 2 years had passed after the autopsy. These results reminded us of the importance of an explanation before and shortly after a forensic autopsy for a better understanding and acceptance by bereaved families.

SIAM-Like phenomenon caused by low doses of alcohol.

BACKGROUND: Swift increase in alcohol metabolism (SIAM) is usually evoked by a large dose of ethanol, which is often demonstrated by an abrupt increase in oxygen uptake. SIAM was induced by low doses of ethanol and evaluated by pharmacokinetic analyses of ethanol and its metabolites. METHODS: Rabbits were initially administered 1.0 g/kg of ethanol solution and the same dose was given to the bolus group 6 hours after the first injection. The infusion group was administered 0.25 g/kg/h of ethanol 2 hours after the first injection. Blood concentrations of ethanol, acetaldehyde, and acetate were then determined and comparisons were made using pharmacokinetic parameters. RESULTS: A significantly higher ethanol elimination rate was observed after re-administration of ethanol to the bolus group. Other pharmacokinetic parameters were unaffected. The concentration at steady state (Css) for the infusion group was stable. A significantly higher level of mean residence time (MRT) in blood acetaldehyde was observed for the bolus group, whereas no MRT changes were observed for the infusion group. A significantly higher level of blood acetate Css was observed after re-administration of ethanol to the bolus group, following the changes in area under concentration and MRT. No Css changes were observed for the infusion group. The Css of acetate at stage 2 was significantly higher for the bolus group, compared to the infusion group. CONCLUSION: Low doses of ethanol enhanced alcohol metabolism in rabbits, according to a pharmacokinetic analysis of circulating ethanol concentrations. Simultaneous analyses of its metabolites followed the kinetic of ethanol.

QT interval dispersion and cardiac sympathovagal balance shift in rats with acute ethanol withdrawal.

BACKGROUND: Dysregulation of autonomic nervous system function and impaired homogeneity of myocardial repolarization are 2 important mechanisms for the genesis of ventricular arrhythmias in nonalcoholic subjects. Our previous study suggested that acute ethanol withdrawal promoted the shift of cardiac sympathovagal balance toward sympathetic predominance and reduced the vagal tone, which were related to a higher incidence of ventricular arrhythmia and related death. However, the homogeneity of myocardial repolarization and its relation with the cardiac sympathovagal balance are unknown, especially in alcoholic subjects. The aim of the present study was to clarify these points. METHODS: Male Wistar rats were treated with a continuous ethanol liquid diet for 49 days, and then subjected to 1-day withdrawal and 1-day withdrawal with 7-day carvedilol (can block the sympathetic nervous system completely via beta1, beta2, and alpha adrenergic receptors) pretreatment. The cardiac sympathovagal balance and homogeneity of myocardial repolarization were evaluated based on the heart rate variability (HRV) and QT interval dispersion (QTd: dynamic changes in QT interval duration). RESULTS: The increase in QTd was observed only in rats at 1-day withdrawal, but not in nonalcoholic, continuous ethanol intake, and 1-day withdrawal with 7-day carvedilol pretreatment rats. At 1-day withdrawal, the low-frequency power/high-frequency power (LF/HF) ratio in HRV was elevated and correlated with the QTd. The increased QTd and elevated LF/HF ratio were normalized by the 7-day carvedilol pretreatment in rats at 1-day ethanol withdrawal. CONCLUSIONS: In rats with an abrupt termination of the chronic continuous ethanol intake, the homogeneity of myocardial repolarization impaired and correlated with the cardiac sympathovagal balance. Carvedilol pretreatment is associated with a reduction in both the QTd and LF/HF ratio, raising the possibility that the cardiac sympathovagal balance shift may be responsible for the impaired homogeneity of myocardial repolarization, and that beta-blocker pretreatment may decrease the mortality risk during alcoholic withdrawal.

Abrupt termination of an ethanol regimen provokes ventricular arrhythmia and enhances susceptibility to the arrhythmogenic effects of epinephrine in rats.

BACKGROUND: Pathologists examining victims of sudden unexpected death encounter alcoholics more often than expected; alcohol may play a role in sudden arrhythmic death. Here we determine whether a pattern of alcohol consumption, chronic ethanol intake, and withdrawal increases the incidence of malignant ventricular arrhythmia and modulates susceptibility to the arrhythmogenic potential of sympathetic stimulation from an epinephrine test in rats. METHODS: Male Wistar rats were treated with a continuous ethanol liquid diet for 7 weeks, and then subjected to 1-day withdrawal or 21-day abstinence. Ventricular ectopy was evaluated by 24-hour electrocardiographic telemetry recording; whole-body sympathetic activation, cardiac sympathovagal balance, and susceptibility to ventricular arrhythmia induced by sympathetic stimulation were evaluated based on blood noradrenalin metabolite concentrations, heart rate variability, and a 3-step epinephrine test. RESULTS: Ventricular arrhythmia and related death were observed only in rats at 1 day of withdrawal, but not in nonalcoholic, continuous ethanol intake or 21-day abstinence rats. One-day withdrawal after a 7-week continuous ethanol regimen elevated circulating noradrenalin metabolite levels and induced cardiac sympathovagal imbalance. Deaths related to the epinephrine test and ventricular arrhythmia induced by low doses of epinephrine were observed only in 1-day withdrawal rats. However, all anomalies were normalized by 21-day abstinence. CONCLUSIONS: Abrupt termination of a 7-week continuous ethanol regimen is sufficient to enhance the whole-body sympathetic activation and cardiac sympathovagal imbalance that contribute to ventricular arrhythmia and sudden death in alcoholic rats. Those providing medical care for alcoholics, including in cases of legal imprisonment, should be aware of the possibility of enhanced susceptibility to sudden arrhythmic death due to the abrupt termination of a chronic ethanol regimen.

Pathological roles of bone marrow-derived stellate cells in a mouse model of alcohol-induced fatty liver.

Chronic alcohol consumption activates hepatic stellate cells (HSCs) and causes fatty degeneration in the liver. However, the origin of HSCs and the mechanism of fatty changes of the liver have not been fully elucidated. Here, we examined the roles of bone marrow-derived cells (BMDCs) in a mouse model with chronic alcohol consumption. We performed bone marrow transplantation from transgenic mice expressing green fluorescence protein (GFP) to female wild-type and ROSA mice (B6.129S7-Gt 26Sor/J, transgenic mice expressing beta-galactosidase, beta-gal) and treated them with ethanol (EtOH) for 8 or 16 wk. GFP-expressing BMDCs increased in the liver with EtOH treatment in a time-dependent manner. In response to excess alcohol consumption, approximately 68% of the BMDCs became activated HSCs in that they expressed alpha-smooth muscle actin. Meanwhile, approximately 67% and approximately 66% of these BMDCs expressed Tnf-alpha and transforming growth factor (Tgf)-beta1, respectively, and the activities were further supported by the excessive mRNA expression of Tnf-alpha and Tgf-beta1 in RT-PCR, respectively. Cell fusion occurs between BMDCs and nonparenchymal cells but scarcely occurs between BMDCs and hepatocytes, demonstrated by double staining of beta-gal/GFP and further supported by the Y-chromosome staining. The EtOH withdrawal normalized most of the abnormalities produced by chronic alcohol consumption. These results indicate that excess alcohol consumption stimulates both the homing of HSCs from the bone marrow and their profibrogenic cytokine production in a mouse model of alcohol-induced fatty liver disease.

Rats in acute withdrawal from ethanol exhibit left ventricular systolic dysfunction and cardiac sympathovagal balance shift.

There is strong evidence that sympathovagal balance plays an important role in the progression of cardiac dysfunction in non-alcoholics. The purpose of this investigation was to determine whether a pattern of continuous ethanol intake and withdrawal modulates the cardiac sympathovagal balance and left ventricular (LV) systolic function in rats. Male Wistar rats were treated with a continuous ethanol liquid diet for 49 days, and then subjected to 1-day withdrawal and 21-day abstinence. Cardiac sympathovagal balance and LV systolic function were evaluated based on heart rate variability (HRV), Western blotting, and echocardiography. Longitudinal data obtained from the same rats showed that the 49-day continuous ethanol treatment induced LV systolic dysfunction, expressed by decreased fractional shortening and ejection fraction. At the 1-day withdrawal, LV systolic dysfunction was deteriorated, and the low-frequency power/high-frequency power (LF/HF) ratio in HRV was elevated because of the depressed HF and the increased LF. Western blot analysis showed an increased expression of myocardial tyrosine hydroxylase and a decreased expression of myocardial acetylcholine. All anomalies were recovered to baseline values with 21-day abstinence. We concluded that acute withdrawal from a 49-day continuous ethanol regimen is sufficient to promote the shift of cardiac sympathovagal balance toward sympathetic predominance and reduced vagal tone, contributing to the further deterioration of LV systolic function in rats. Those providing medical care for alcoholics should be aware of this enhanced susceptibility to LV systolic dysfunction with abrupt termination of a continuous ethanol regimen.

Legal medicine and the death inquiry system in Japan: a comparative study.

In Japan, there are two different systems of death investigation: criminal inspection and judicial autopsy from a criminal justice standpoint and, from a public health standpoint, administrative inspection and either administrative or consent autopsy. As a result, it is sometimes unclear which system is responsible for investigating a death. In this study, the Japanese death inquiry system is compared with the coroner's system used in some other countries. One aim of a death inquiry is to prevent future loss of life in similar circumstances, which the Japanese system does not consider. The systematic reform of the Japanese death inquiry system is thought to be necessary.