Stephan Seidl

Phosphatidylethanol in Human Organs and Blood: A Study on Autopsy Material and Influences by Storage Conditions.

OBJECTIVE Phosphatidylethanol (PEth) is an abnormal phospholipid that is formed and accumulated in mammalian cells that have been exposed to ethanol. PEth has been proposed as a marker of ethanol abuse. This study was conducted to investigate the concentration of PEth in blood and organs obtained during the autopsy of alcoholics. In addition, we performed experiments on rat tissues and human blood to evaluate the effect of various storage conditions on PEth concentrations. METHODS Human tissues and blood from alcoholics and controls were obtained at autopsy and frozen at -20 degrees C until extraction. Blood from healthy donors was incubated with ethanol for 24 hr and thereafter either extracted directly or stored at room temperature, stored at 4 degrees C, frozen at -20 degrees C, or frozen in liquid nitrogen and stored at -80 degrees C before extraction. Rats were given intraperitoneal injections of ethanol and then killed, either while still intoxicated or when sober. Rat organs were homogenized and extracted directly, after a period of storage, and/or after freezing at -20 degrees C. PEth concentration was analyzed using HPLC and verified by mass spectrometry. RESULTS In all rat organs studied, PEth was formed during freezing at -20 degrees C with ethanol present. PEth concentrations of 9 to 205 mumol/liter were observed in the blood obtained at autopsy. The highest value was found in the case with the highest blood alcohol concentration (114 mmol/liter) at the time of death. In the experiments on human blood stored with ethanol present, PEth concentrations were not affected after 72 hr at 4 degrees C or after freezing in liquid nitrogen and storage at -80 degrees C for up to 144 hr but were slightly elevated after 24 hr at room temperature and at -20 degrees C. PEth was found in all organs obtained from the cadavers of alcoholics. Storage of organs at 4 degrees C for 24 hr with ethanol present had no effect on the PEth concentration. The PEth concentration was unaffected when no ethanol was present at the time of freezing. CONCLUSIONS The rat experiments indicated that the very high PEth concentrations found in the organs of the alcoholics were probably largely formed while the organs were frozen at -20 degrees C. Our data suggest that tissue material from bodies that were exposed to ethanol must be stored properly to obtain reliable results from subsequent analysis for PEth. Tissue should not be frozen at -20 degrees C but instead stored refrigerated until extraction, preferably within hours of autopsy, or frozen in liquid nitrogen and stored at -80 degrees C. Blood samples that contain ethanol can be stored refrigerated for up to 72 hr or frozen in liquid nitrogen and stored at -80 degrees C without affecting PEth levels.

Ethyl glucuronide-a biological marker for recent alcohol consumption.

Ethyl glucuronide (EtG) is a non‐volatile, water‐soluble metabolite of ethanol, with a high storage stability. It can be detected in body fluids, tissues, sweat and hair for an extended time period after the elimination of ethanol from the body. EtG closes the gap between short‐term markers for alcohol consumption such as ethanol or methanol and long‐term markers for alcohol misuse such as GGT, MCV and CDT. Due to its specific time‐frame of detection and its high sensitivity and specificity, EtG is a promising marker for alcohol consumption and for relapse control that enables the therapist to intervene at an early stage of relapsing behaviour. The aim of this review is to give an overview of analytical techniques for the detection of EtG, its clinical use and remaining questions.

Ethyl glucuronide: on the time course of excretion in urine during detoxification.

Ethyl glucuronide (EtG) is a promising new biological state marker of recent alcohol consumption that detects alcohol use reliably over a definite time period. Other currently available markers lack acceptable sensitivity and specificity. Our aim is to elucidate under naturalistic conditions the time course of EtG excretion in urine following alcohol consumption and to show how this can be utilized for monitoring and prognosis in patients. There are no other existing data on this issue to date. One hundred and thirty‐eight urine samples from 28 male alcohol withdrawal patients were drawn every 3‐24 hours for up to 94 hours after hospitalization. Breath ethanol concentration (mean) at hospitalization was 900 mg/L. Patient age in years was 40.3 (mean). Determination of urine EtG was performed by gas chromatography/mass spectrometry (GC/MS) with deuterium‐labelled EtG as an internal standard. The strongest correlations (p<0.01) were found between EtG determinations in the different patient when breath ethanol concentrations (BEC) were 0 and 48 hours after BEC=0 (r=0.747), EtG 24 and 48 hours after BEC=0 (r=0.872), and in the time frame of detection (hours) of EtG and EtG 48 hours after BEC=0 (r=0.762). No significant correlation was found (Mann‐Whitney test) between EtG concentrations in urine at different time points between the groups of patients with (a) 1 or less‐2, (b) 3‐4 or more previous hospitalizations, (c) a history of seizures (yes/no) or (d) an age above or below the median (40.5). EtG excretion in urine is not random, but seems rather to follow a kinetic profile. Furthermore our preliminary data indicate, that there is no significant difference for EtG concentration in urine when correlated to group variables such as age, seizures and hospitalizations.

The direct ethanol metabolite ethyl glucuronide: A specific marker of recent alcohol consumption

In clinical as well as in forensic practice, occupational medicine, at court, for both traffic safety and safety at workplaces biological state markers of high sensitivity and specificity, capable of monitoring those in treatment for alcohol dependence or poly-drug-abusers as well as social drinkers in risky situations (driving, workplaces) are required. The known markers can not be considered to be satisfying with regard to these parameters. Furthermore, they do not cover the whole time axis for alcohol intake — i.e. there is a gap with regard to alcohol consumption of between 1 day and 1 – 2 weeks between short- and long-term-markers like ethanol, methanol, HTOL/HIAA ratio and CDT, GGT or MCV respectively. These traditional markers are in many cases besides alcohol influenced by age, gender and a variety of substances and non-alcohol-associated diseases. Ethyl glucuronide (EtG) is a non-volatile, water-soluble, stable upon storage, direct metabolite of ethanol with a molecular weight of 222 g/mol that can be detected in body fluids, tissues and hair. With its specific time frame of detection intermediate between between short- and long term markers and a high sensitivity and specificity, ethyl glucuronide is a promising marker of alcohol consumption in general that can be detected for an extended time period after the complete elimination of alcohol from the body and a marker for relapse control that enables the therapist to intervene at an early stage of relapsing behaviour.

Pathological Features of Death From Lightning Strike

Lightning strikes cause more deaths in the United States than other natural disasters, such as hurricanes, tornadoes, volcanoes, and floods. Lightning is a transfer of an electrical charge and results from the sudden environmental discharge of static electricity. The power of lightning is estimated to be between 10,000 and 200,000 A of current, with estimated voltage ranging from 20 million to 1 billion V. The effects of lightning on the human body depend on a number of features, such as the intensity of the current, the time it spends passing through the body, the pathway involved, the activity and position of the person at the time of the event in relation to the ground, and the kind of strike (direct strike, contact voltage, side splash, ground strike, or wire-mediated lightning). Lightning strikes result in multisystem dysfunction, and survivors may experience prolonged disability following recovery from the initial insult. Electrical energy causes muscular spasm and necrosis, thrombosis, blood vessel tears, unconsciousness, and motor and sensory function abnormalities. Most deaths after lightning strikes occur either because of primary cardiac arrest or hypoxia-induced secondary cardiac arrest. If multiple persons are struck, attention at the scene should be directed first to those who appear dead, because they may be in respiratory arrest and in urgent need of immediate cardiopulmonary resuscitation that can be successful in lightning strike victims for far longer than would seem reasonable in other types of injury.