Cyanide concentrations in blood and tissues of fire victims.

Cyanide poisoning has been regarded to contribute the fatal outcome in fire victims. The toxicity of inhaled hydrogen cyanide (HCN) at the cellular level was evaluated considering the impact of methemoglobin (MetHb) produced by fire gases. Cyanide (CN) concentrations and total hemoglobin contents were measured in right heart blood (RHB) and seven organs/tissues (basal ganglia, brain stem, heart, lung, liver, kidney and psoas muscle) collected from 20 fire fatalities. MetHb and carboxyhemoglobin saturations were also measured in RHB. The amount of CN probably bound to the cytochrome c oxidase of the tissue cells (CCO-CN) was extrapolated from CN and hemoglobin contents in RHB and organs/tissues, MetHb saturation in RHB and binding capacity of MetHb for CN. CN concentrations in RHB showed a wide range with the highest concentration of 8.927 µg/mL. The lung contained the largest CN content among organs/tissues with the mean concentration of 2.219 µg/g, then the heart (0.259 µg/g) and it was lower than 0.100 µg/g in others. Exceedingly large amount of CN in the lung could be explained by high hemoglobin content, being the port of entry of HCN and postmortem diffusion of fire gases. CCO-CN was theoretically present in about 20% of organ/tissue samples, most commonly in the basal ganglia (10 samples, with the mean of 0.059 µg/g) followed by heart (eight samples, with the mean of 0.109 µg/g). No CCO-CN was found in liver and kidney. HCN might have the effect on brain and heart.

Detection of abused drugs in human blood by using the on-site drug-screening device Oratect® III.

A simple and precise drug screening method was developed for the detection of abused drugs in whole blood by using the Oratect® III device that is usually employed for the detection of drugs in saliva. Whole blood was acidified with phosphoric acid, following which the hemolyzed solution was filtered through the ultrafiltration column Vivaspin 2 Hydrosart®. The filtrate was then tested for the presence of drugs using Oratect III. The detection limit of the device for methamphetamine, amphetamine, morphine, codeine, dihydrocodeine, diazepam, alprazolam, estazolam, and prazepam in whole blood was 125, 125, 50, 50, 50, 25, 60, 15, and 75ng/mL, respectively. The concentration range detected was between therapeutic and toxic drug levels; therefore, the proposed method can be applied for detecting the presence of abused drugs in blood. Our method is a novel, optimized technique for use in forensic laboratories to screen whole blood for drugs of abuse.

Sweet taste-induced analgesia: an fMRI study.

We investigated the brain activation associated with sweet taste-induced analgesia by 3-T functional magnetic resonance imaging, the mechanism of which is considered to involve the central nervous system. After 12 healthy individuals ingested tasteless gelatin (nonsweet condition) or sweet glucose (sweet condition) in a magnetic resonance imaging scanning gantry, the cold pressor test was applied to their medial forearm. Under both conditions, the cold pressor test robustly activated the pain-related neural network, notably the anterior cingulate cortex, insula, posterior parietal cortex, and thalamus, although such activations under the sweet condition weakened with pain threshold increase, compared with those under the nonsweet condition. Together with emotional changes in pain appraisal, our findings provide objective representation of sweet taste-induced analgesia in the human brain.

Methamphetamine protects against stress-induced gastric mucosal lesions in mice.

Stress-induced gastric mucosal injury is a common clinical entity. On the other hand, abuse of methamphetamine (MA) represents a growing social problem. MA users are frequently in stressful situations. In this study, we examined the effects of MA on gastric injury, corticosterone level and immunomodulation using a water-immersion restraint stress (WRS) mouse model that is well known to induce gastric lesions. Mice were randomly divided into five groups: (1) the normal group, (2) the 3 hour (3 h)-WRS group, (3) the 6 hour (6 h)-WRS group, (4) the MA (3 mg/kg) plus 6 h-WRS group and (5) the MA (30 mg/kg) plus 6h-WRS group. As expected, most animals examined (above 90%) showed gastric injury after the WRS exposure. However, administration of MA at both 3 and 30 mg/kg resulted in significant suppression of the injury. The corticosterone levels were increased by exposure to the stress and/or MA, but there was no difference between these groups. The levels of the serum cytokines IL-6, IL-10 and TNF were increased by WRS, and were markedly increased by MA plus WRS; in particular, the level of IL-6 was synergistically increased. On the contrary, the level of IL-1beta was significantly decreased by WRS and MA plus WRS. This is the first report showing the protective effect of MA on stress-induced gastric injury, although further study is necessary to resolve the mechanism of MA-driven suppression of the injury.

Urine levels of drugs for which Triage DOA screening was positive.

The purpose of this study was to investigate the relationship between urine levels of target drugs of abuse for which Triage DOA gave positive results, as well as the cut-off levels for these drugs. Thirty-eight forensic urine samples positive for commonly abused drugs were involved. Of these samples, 12 were positive for barbiturates (BAR), 11 for benzodiazepines (BZO), 8 for opiates (OPI), 7 for amphetamines (AMP), and 4 for tricyclic antidepressants (TCA). In the BAR-positive urine samples, phenobarbital, amobarbital or barbital was detected at concentrations higher than cut-off levels. In the BZO-positive samples, diazepam, nordiazepam, triazolam, nitrazepam and/or midazolam was detected at concentrations lower than cut-off levels; in the triazolam-involved urine, alpha-hydroxytriazolam, a metabolite of triazolam, showed concentrations higher than cut-off level. In the AMP-positive samples, methamphetamine was detected at concentrations higher than cut-off level. Urine samples positive for OPI contained total dihydrocodeine, codeine or morphine at concentrations higher than cut-off levels. In TCA-positive samples, amitriptyline was detected at concentrations higher or lower than cut-off level, and clomipramine was detected at a concentration much lower than cut-off level. Metabolites of BZO and TCA, which are not typically analyzed by instrumental procedures, may cross-react to varying degrees with the antibodies used for Triage DOA.

Ethanol consumption alters expression and colocalization of bile salt export pump and multidrug resistance protein 2 in the rat.

Chronic ethanol consumption elicits detrimental changes of liver metabolism. By employing a 12-week-long feeding regimen, we investigated the effects of chronic ethanol consumption on the expression and localization of bile salt export pump (Bsep), a major canalicular exporter of bile salts, and multidrug resistance protein 2 (Mrp2), a canalicular organic anion transporter, in the rat liver. RT-PCR, confocal immunofluorescence microscopy, immunoblotting, and quantitative colocalization analysis were used to examine their gene and protein expression, and changes in the distribution of antigenic sites. Bsep mRNA was upregulated, while Mrp2 mRNA responded by downregulation. In agreement with mRNA, the expression of Bsep protein increased, while the expression of Mrp2 protein responded with a decrease, suggesting that the expression of both of them is transcriptionally regulated. Confocal immunofluorescence microscopy showed disruption of the colocalization of Bsep and Mrp2 proteins at the hepatocyte canalicular membrane and their relocation intracellularly. Quantitative colocalization analysis of Bsep and Mrp2 proteins revealed a steady decrease in the degree of colocalization and Mrp2 expression, indicating that although the properties of both transporters are affected, Mrp2 is altered more. These findings provide evidence that ethanol alters Bsep and Mrp2 canalicular transporters in the rat liver, at both the mRNA and protein levels. Mrp2 shows deeper involvement. Eight weeks appears to be a critical time point in this process.

Forensic aspects of complications resulting from cardiopulmonary resuscitation.

While cardiopulmonary resuscitation (CPR) can save lives, it can also injure patients. As a result, forensic pathologists often see CPR-related injuries during autopsies that are unrelated to the patients' primary cause of death. Therefore, pathologists must be able to distinguish between CPR-related injuries and those caused by other factors, such as assaults or accidental violence. This distinction is complicated because even therapeutically unimportant injuries can have forensic significance. For example, resuscitative injuries are observed frequently in the neck and the chest. This article focuses mainly on complications due to ventilation and chest compression during CPR. The following iatrogenic complications are described: bruising and abrasions in the face and neck, fractures of the hyoid bone and thyroid cartilage, air way injuries, vomitus aspiration, positional error of the tube for intra-tracheal intubation, petechiae, retinal hemorrhages, subarachnoid hemorrhages, rib and sternum fractures, bone marrow embolism, cardiac injuries including myocardial hemorrhages and frothy heart blood, and injuries to the abdominal organs such as liver and spleen.

Nicotine and cotinine levels in body fluids of smokers who committed suicide.

Cigarette smoking is associated with a higher risk for suicide. The present study was conducted on the hypothesis that suicide smokers show higher nicotine and cotinine levels in blood and urine than non-suicide smokers. We determined nicotine and cotinine levels in blood and urine of 87 deceased individuals (18 suicides and 69 non-suicides) by gas chromatography. The smoking rate was 77.8% for individuals who committed suicide and 42.0% for those who did not commit suicide. Average nicotine and cotinine levels in blood were significantly higher in the suicide smokers than in the non-suicide smokers (nicotine: 93.2+/-46.6 ng/ml versus 25.8+/-14.4 ng/ml, p<0.0001 and cotinine: 378+/-235 ng/ml versus 201+/-137 ng/ml, p<0.005). Average levels of urinary nicotine and cotinine were also significantly higher in the suicide smokers than in the non-suicide smokers (nicotine: 1980+/-2210 ng/ml versus 394+/-376 ng/ml, p<0.005 and cotinine: 1170+/-1330 ng/ml versus 414+/-290 ng/ml, p<0.05). Twenty-six decedents were intoxicated with alcohol, and they included 7 suicides (7 smokers) and 19 non-suicides (15 smokers). Our data suggest that cigarette smokers who commit suicide smoke more heavily than other cigarette smokers.

Effects of drinking and smoking on endogenous levels of urinary gamma-hydroxybutyric acid, a preliminary study.

The aim of this study was to determine if the endogenous levels of gamma-hydroxybutyric acid (GHB) in urine were affected by drinking and smoking. Urine samples were obtained from 20 healthy volunteers (15 males, 21-45 years; 5 females, 22-24 years). This population included four average drinkers (males), 4 average smokers (males), and 12 nonsmokers/nondrinkers (seven males and five females). Urinary levels of GHB were measured by gas chromatography. No gender differences were observed in the urinary levels of endogenous GHB. The urinary levels of GHB in males were 0.52+/-0.37 microg/ml in smokers, 0.28+/-0.21 microg/ml in nonsmokers/nondrinkers, and 0.23+/-0.04 microg/ml in drinkers. Urinary GHB levels were measured three times a day for 5 consecutive days in a male from each group. Large intra-individual differences were observed over the 5-day period in a smoker and a nonsmoker/nondrinker. No significant changes in daily endogenous GHB levels were observed in a drinker during the period. Our preliminary results suggest that stimulatory effects of nicotine on the central nervous system (CNS) may result in an increase in nocturnal formation of GHB and the depressive effects of ethanol on the CNS may not affect, even may inhibit, nocturnal production of GHB.

[Nicotine and cotinine levels in body fluids of habitual smokers who committed suicide].

Suicide is a serious social problem because over 30,000 people commit suicide every year since 1998 in Japan. Cigarette smoking is associated with a higher risk for suicide and attempted suicide. We determined nicotine and cotinine levels in blood and urine of 104 deceased individuals (21 suicides and 83 non-suicides). Of the 21 suicides, 16 (76.2%) were smokers; the smoking rate in non-suicides was 41.0% (34 persons). Average levels of nicotine and cotinine in blood were significantly higher in the suicide smokers than in the non-suicide smokers (nicotine: 95.6 +/- 43.9 ng/ml vs. 28.0 +/- 15.2 ng/ml, p < 0.0001; and cotinine: 385 +/- 220 ng/ml vs. 229 +/- 181 ng/ml, p < 0.02). Average levels of nicotine and cotinine in urine also significantly higher in the suicide smokers than in the non-suicide smokers. There were eight patients with psychiatric diseases such as schizophrenia, depression and alcohol dependence. Of the eight patients, four were suicide smokers; only a person used antipsychotics. Thirty-one alcohol-intoxicated decedents consisted of 8 suicides (8 smokers) and 23 non-suicides (17 smokers). Our data demonstrate that there is a marked increase in cigarette smoking in habitual smokers with psychiatric disorders before committing suicide. Quantitatively monitoring the severity of stress using blood nicotine level may enable physicians more objectively to find out nicotine dependents who are in the state of an imminent suicide attempt and timely to administer medical treatment for preventing suicide.

Effects of perimortem physical factors associated with death on exogenous ethanol concentrations in cardiac blood.

The purpose of this study was to determine whether perimortem physical factors associated with death affect exogenous ethanol concentrations in cardiac blood. Forty-one autopsies of alcohol-intoxicated decedents with no or little putrefaction were involved. Postmortem intervals ranged from 6 to 96 h at the time of autopsy. Our 41 cases consisted of 17 fire victims, 8 drowned persons, 3 blood-loss patients who underwent unsuccessful cardiopulmonary resuscitation (CPR) for 30-50 min, 3 blood-loss persons without CPR, three non-blood-loss patients who underwent unsuccessful CPR for 5-60 min, and seven people who died of other causes. Ethanol concentrations in right cardiac chamber blood ranged from 0.12 to 4.40 mg/g (mean 1.39 mg/g) in all cases. An excellent correlation was observed between ethanol concentrations in right cardiac chamber blood and femoral venous blood (y=1.01x-0.087, n=38, r2=0.990). The ratios of blood ethanol concentrations in the left to right cardiac chambers were significantly lower in blood-loss decedents with CPR (0.71+/-0.13) than other decedents (0.92+/-0.07-1.06+/-0.06). Two factors, blood-loss and prolonged chest compression under ventilation, might have caused a decrease in blood ethanol concentration in the left cardiac chambers. Postmortem cardiac blood ethanol concentration as well as postmortem femoral venous blood ethanol concentration may usually reflect blood ethanol levels at the time of death when putrefaction is not evident. However, special attention is required for cases that received CPR after massive bleeding.

A fatal poisoning caused by methomyl and nicotine.

A 35-year-old male was found lying in a prone position in his room. He was in cardiopulmonary arrest on arrival to hospital and was pronounced dead. There was no attempt at resuscitation. No miosis was observed on admission. At post-mortem his stomach contained 170 g greenish liquid with a small amount of shredded tobacco leaves. The serum cholinesterase activities were 47-90 IU (normal range for male: 200-440 IU). GC and GC-MS analyses showed nicotine (21.8 mg), methomyl (304 mg), and triazolam (1.69 mg) in his stomach. He had consumed tobacco leaves, Lannate containing water soluble methomyl (45%), and Halcion tablets containing 0.25 mg triazolam. Methomyl concentrations in blood were 3-8 ng/ml. Substantial amounts of methomyl (2260-2680 ng/ml) were detected in cerebrospinal fluid and vitreous humor. Nicotine concentrations in blood ranged from 222 to 733 ng/ml. A small amount of triazolam was detected only in bile (176 ng/ml) and liver (23 ng/g). The cause of death was respiratory paralysis produced by the additive effects of methomyl and nicotine shortly after consumption.