Site-dependent postmortem changes in blood cocaine concentrations

When a forensic toxicologist interprets postmortem blood cocaine findings he usually must make assumptions regarding perimortem drug concentrations. In-vitro studies have shown that cocaine rapidly hydrolyzes in unpreserved blood, particularly at elevated temperatures. However, other studies have demonstrated site-dependent postmortem release of some drugs from tissue stores accompanied by increases in drug concentrations in the blood. This study was undertaken to investigate whether blood cocaine concentrations change in the body during the postmortem interval and, if so, to measure the direction and magnitude of the changes. In medical examiner cases in which scene investigation suggested that the decreased was a cocaine user, blood samples were collected as soon after death as possible. At autopsy, a second set of samples was collected. Analysis of paired samples by gas chromatography/mass spectrometry (GC/MS) revealed dramatic differences in the cocaine concentration. The magnitude and direction of the change appears to be site dependent. Usually, but not invariably, cocaine concentration in subclavian vein blood decreases while that in heart, aorta, and femoral vein blood increases during the interval between death and autopsy. The findings emphasize the danger inherent in attempting to estimate the concentration of cocaine in blood at the time of death from postmortem data.     

The use of absolute refractory period in the estimation of early postmortem interval

The estimation of the time since death (postmortem interval) is one of the most difficult problems in forensic pathology. Most methods currently employed use temperature-based algorithms intended to model the cooling of the body after death and thus estimate the postmortem interval. These methods are subject to considerable inaccuracy but their reliability can be improved if a range of other observed criteria such as lividity and rigor are also taken into consideration. The aim of the present study was to investigate the feasibility of using the absolute refractory period as an adjunct to the estimation of postmortem interval. The relationship between the `postmortem interval' and the `duration of absolute refractory period' was investigated using the rat sciatic nerve. A strong correlation between the duration of the absolute refractory period and the postmortem interval was observed. When both absolute refractory period and temperature were used in conjunction, the strength of this correlation was increased.     

Postmortem changes in resistivity of the anterior abdominal wall during the early postmortem period in rats

An accurate and reproducible technique was employed for the determination of the resistivity of excised portions of the anterior abdominal wall of rats. Resistivity decreased linearly (r = −0.93; P < 0.001) from 1438 ± 131 Ω · cm, immediately following death, to 360 ± 144 Ω · cm at a postmortem interval of 36 h. These changes are believed to reflect the morphological and/or chemical changes which occur at the cellular level during the early postmortem period.     

Postmortem changes in electrical resistance of the gastric wall during the early postmortem period in rats.

A simple and reliable technique was employed for measurement of (dc) electrical resistance of the gastric wall of rats. Tissue resistance decreased linearly (r = -0.87; P less than 0.001) from 66.1 +/- 12.1 kOhm/cm2 immediately following death to 23.3 +/- 3.5 kOhm/cm2 at a postmortem interval of 24 h. These changes are believed to reflect progressive postmortem changes in the physical dimensions of the intercellular space and/or changes in electrical conductivity of the intercellular/extracellular fluids.     

Interleukin-6 and C-reactive protein serum levels in sepsis-related fatalities during the early postmortem period

Postmortem interleukin-6 (IL-6) and C-reactive protein (CRP) serum levels were investigated prospectively in sepsis-related fatalities and non-septic fatalities by using a linear regression model. At least three blood samples were collected between 0.3 and 139 h postmortem from sepsis-related fatalities (n=8) and non-septic fatalities (n=16). In addition, one antemortem blood sample was collected shortly before death from the septic patients. Antemortem and postmortem IL-6 and CRP levels were highly elevated in all individuals included in the sepsis group. An excessive postmortem increase of IL-6 serum levels associated with progressive time after death was observed in five out of the eight septic patients. Both, IL-6 and CRP serum concentrations seem to be suitable biochemical postmortem markers of sepsis. The determination of IL-6 serum levels above 1500 pg/ml in peripheral venous blood obtained in the early postmortem interval can be considered as a diagnostic hint towards an underlying septic condition. A more precise postmortem discrimination between sepsis and non-septic underlying causes of death is provided by the postmortem measurement of serum CRP in peripheral venous blood: on condition that at least two postmortem CRP values have been determined at different time points postmortem, the CRP level of a deceased at the time of death can be calculated by using linear regression analysis. When assessing postmortem IL-6 and CRP concentrations as biochemical postmortem markers of sepsis, various clinical conditions, such as a preceding trauma or burn injury going along with elevated IL-6 and/or CRP levels prior to death as a result of the systemic inflammatory response syndrome (SIRS) should be taken into consideration, thus adding relevant information for the practical interpretation of the results.     

Self-extraction of teeth involving gamma-hydroxybutyric acid

A case involving self-extraction of teeth linked to the abuse of gamma-hydroxybutyric acid (GHB) is reported. A 28-year-old woman and her 29-year-old boyfriend were discovered by paramedics following an extensive period of GHB use. The paramedics were alerted by a neighbor who had heard screaming from the house. On presentation to the accident and emergency department, it was noted that the female had 18 fresh extraction sockets visible intra-orally. At the scene, a mirror, a pair of pliers, and a bowl containing human teeth were found. Charges of assault were taken to the courts against the boyfriend who was subsequently acquitted. Odontological evidence centered on whether or not it was possible to self-extract the teeth using the pliers found. This case is the first to describe possible oral self-mutilation under the influence of GHB and odontologists should always consider self-injury as an explanation for intra- and perio-oral injuries of unknown origin.     

Evaluation of histologic changes of the skin in postmortem period

ABSTRACT: Determination of the time of death is an important consideration in forensic practice. Many methods have been attempted to accurately and systematically determine the postmortem interval (PMI). Histologic examination of the skin or appendages is one of the methods tried by few researchers. However, no attempt had been made to analyze the histologic changes in the skin and appendages simultaneously and to compare them with PMI. We sequentially studied the histologic changes of the skin and appendages in the early PMI. The results of the present study show that the skin undergoes progressive morphological changes in the postmortem period. The epidermis and the dermis appeared normal for 6 hours after death, and after this period, degenerative changes began. By 6 to 9 hours after death, degeneration began in the dermis, and by the end of 18 hours, the dermis began to disintegrate. The sweat glands appeared normal for approximately 3 to 4 hours. For 18 hours after death, the sebaceous glands and hair follicles appeared normal, and after this period, degeneration began.     

GC-MS determination of gamma-hydroxybutyric acid (GHB) in blood

A fast and efficient procedure has been developed for the analysis of total mercury in human tissues and blood using a hydride vapor generator system coupled to an atomic absorption spectrometer (HVG-AA). Tissue and blood samples were digested in a pressurized microwave decomposition system and the digest diluted prior to formation of free mercury vapor and analysis by atomic absorption. Recovery studies performed on 10 spiked/unspiked pairs of human liver and on 10 spiked/unspiked pairs of human blood samples yielded average recoveries of 99.7% (CV=0.4%) and 101.2% (CV=0.5%), respectively. The method detection limit for liver and blood was 50 microg Hg/kg and 12.5 microg Hg/l, respectively. The "normal" concentrations of mercury in human liver and blood are 33-490 microg Hg/kg and 0.6-59 microg Hg/l, respectively [1]. This method is able to determine mercury poisoning levels and may also be applied to detect mercury near the lower levels of these "normal" ranges, using the standard addition method approach.     

Stability of plasma total cholesterol, triglycerides, and apolipoproteins B and A-I during the early postmortem period

The stability of plasma lipids and apolipoproteins during the early postmortem period was studied by taking four duplicate blood samples from eight cadavers 2, 6, 12, and 24 h after death. The bodies were kept at +4 degrees C. The plasma samples were analyzed for total cholesterol (TC), triglycerides (TG), apolipoprotein B (apo B), and apolipoprotein A-I (apo A-I). In TC, values rose by 6 and 11% in two cases, and in six cases diminished 3 to 15% during the first 6 h compared to values obtained 2 h postmortem. The greatest changes were a continuing rise in one case by 33% and a fall by 21% in another case during 24 h. In TG values marked changes took place including one case with a rise of 67% within 24 h. The concentrations of apo B rose by 9 to 11% in three cases and fell by 3 and 4% in two cases during 6 h, but during the whole study period a rise up to 78% occurred. In the concentrations of apo A-I, cases fell by as much as 42% in 6 h, and in one case rose by 20% during 6 h. The results indicate that unpredictable fluctuations occur in plasma lipid and apolipoprotein values within 24 h after death, and they should be interpreted cautiously if the samples have been taken after a prolonged postmortem period.     

Heart fatty acid binding protein as a marker for postmortem detection of early myocardial damage

Depletion of heart fatty acid binding protein (H-FABP) from cardiomyocytes with varying post-ischemia intervals was studied in acute myocardial infarction (AMI) model of rat, and 22 human autopsy cases were studied with streptavidin-peroxidase conjugated method (S-P). It was observed that as early as 15 min after ischemia, the depletion of H-FABP could be detected in model rats. With the ischemic time prolonged, the depletion of H-FABP was more and more evident. In all human cases with myocardial infarction, absent H-FABP staining could be found in infarcted area. And in some suspected early myocardial infarction cases, depletion of H-FABP staining could be demonstrated in areas that showed normal hematoxylin-eosin (HE) staining. The blood samples from model rats before ligation, at varying post-ischemia intervals and various postmortem time were measured for plasma concentration of H-FABP with enzyme-linked immuno-sorbent assay (ELISA) method. At 15 min after myocardial ischemia, the concentration of H-FABP was 4 times higher (546.0+/-85.3 microg/l) than that of the baseline level (103.7+/-94.1 microg/l). With the continuation of ischemic time, the concentration of H-FABP increased and peaked at 4 h (1953.5+/-405.3 microg/l), then decreased. The plasma concentration of H-FABP decreased slightly with postmortem time, but was still significant higher at any postmortem intervals than that of baseline level within 48 h after death. The results suggest that H-FABP staining can detect very early ischemic damages in human myocardium and the elevated plasma concentration of H-FABP in rat was an indicator of AMI, which was not affected by autolysis.     

超生反应阶段人骨骼肌对电刺激反应的形态学研究

利用人骨骼肌在死亡早期阶段能对电刺激产生兴奋和收缩的特点，以加大刺激强度为手段，对18例死亡早期阶段（1小时25分至6小时40分）具有超生反应能力的尸检骨骼肌和17例稍晚阶段（10～60小时）不具有超生反应能力的尸检骨骼肌，进行了电刺激损伤的系列形态学对比研究。结果表明，死亡早期阶段的肢体骨骼肌，经电刺激后，能产生整个肌肉或局部肌肉的收缩；在电极周围以及电路中的肌纤维形成大片的极具特点的收缩带、条带状嗜酸性变、高度变形的节段状和盘片状崩解等改变；稍晚期阶段受刺激的肌纤维则未见上述改变。探讨形态学改变的形成机理及意义，为研究死亡早期阶段的电损伤提供形态学基础。     

Determination of endogenous gamma-hydroxybutyric acid (GHB) levels in antemortem urine and blood

Gamma-hydroxybutyric acid's (GHB's) natural presence in the body has made the interpretation of its levels a challenging task for the forensic toxicologist. This study was designed to measure endogenous GHB levels in antemortem urine and blood samples. The range detected in urine was from 34 to 575 microg/dl and in blood from 17 to 151microg/dl. The results indicate that the concentration of endogenous GHB in urine and blood concur with the suggested cut-off levels at 1000 and 500 microg/dl, respectively.     

Brain amino acid abnormalities in liver disease--a postmortem study

In a postmortem exploratory study, we examined whether specific amino acid abnormalities associated with liver diseases in vivo may also be detected in human brain samples obtained at clinical autopsies. The branched-chain amino acids (BCAA: valine, leucine, isoleucine) were decreased in the group of patients with liver diseases compared with the control group, whereas the aromatic amino acids (AAA: phenylalanine, tyrosine) were increased. However, the ranges overlapped significantly and were not statistically different. The molar ratio BCAA/AAA was determined to be 1.92 in the collection of patients with liver diseases compared with 2.27 in the control group. In patients with liver disease, ornithine concentrations in the brain appeared significantly decreased whereas glutamine was significantly increased. No significant difference was found in the brain concentrations of proline. Amino acid analysis may contribute to the understanding of pathophysiological mechanisms of liver disease, which are discussed, and may supplement the postmortem diagnosis.     

Chemical composition and structure of the microcrystals formed between silver(I) and gamma-hydroxybutyric acid and gamma-hydroxyvaleric acid

This study examined microcrystals formed by silver with gamma-hydroxybutyric acid (GHB) and gamma-hydroxyvaleric acid (GHV), the five-carbon analog of GHB, in the presence of silver, copper, and lanthanide nitrates. Distinct microcrystals formed with silver (+1) and lanthanum (+3) ions but not with the copper (+2) ions. The crystals formed with GHB were distinctly different than those formed with GHV and in all cases, the drug microcrystals were easily distinguishable from reagent crystals. X-ray diffraction analysis provided definitive structure for the microcrystals. The morphological differences between the silver-GHB and silver-GHV crystals were characterized using simple measurements such as size and angles provided by image recognition software. The utility of the test for casework was demonstrated using spiked beverage samples.