Postmortem Aortic Dissection: An Artifact of the Embalming Process

Aortic dissection (AD) is a serious condition that affects 3/100,000 individuals a year. Recently, a case report was published describing an embalmed patient with an aortic dissection. The purpose of this study was to examine the frequency of AD among 80 embalmed cadavers and confirm the AD with histopathologic evaluation. In seven cases of grossly identified AD, six were determined to be due to the embalming procedure and only one case of true antemortem AD was confirmed. These results suggest that aortic morphology can be altered by administration of the embalming fluid and that alterations can mimic AD, not only on gross inspection but also on postmortem imaging. Awareness of this embalming artifact may prevent misdiagnosis of an aortic dissection in an embalmed patient, a point particularly useful for autopsy pathology that may include postmortem examination of an embalmed patient.     

Detection of ketosis in vitreous at autopsy after embalming

Ketosis occurs in ketoacidosis or malnourishment. When either is suspected in relation to a death, it may be important to analyze for ketosis at autopsy. We encountered a case where starvation was suspected in a deceased nursing home resident, where the body had been embalmed prior to autopsy. Gas chromatography (GC) was unable to separate acetone from formaldehyde, a component of embalming fluid. The Acetest is a simple test that can detect acetone and acetoacetate in body fluids. We validated the Acetest with GC on vitreous. The Acetest and GC were consistent except at very low levels of acetone or acetoacetate. The sensitivity of the Acetest for acetoacetate in vitreous was 10 mg/dL, consistent with early starvation. Significant interference from embalming fluid did not occur. The Acetest was negative in the described case. The Acetest is a simple and useful test for the detection of ketosis in embalmed autopsies.     

Guidelines for the recognition of cemetery remains in Greece

Forensic pathologists frequently consult anthropologists for the identification of skeletonized human remains. These remains may be the result of criminal activity or remains that were unearthed because of erosion, or during construction projects. In some cases, human remains that had been previously buried in a cemetery may be the subject of a forensic investigation. Early recognition of cemetery remains prevents unnecessary efforts and conserves precious resources. One of the key characteristics of cemetery remains is the presence of embalmed tissue. However, there are countries where embalming is not a common practice, and other clues must be sought for identifying previously buried remains. Current funerary customs in Greece and, in particular, the tradition of exhumations result in a large number of misplaced human remains. The present study presents examples of cemetery remains from Greece and offers guidelines for recognizing changes on skeletal remains that may be indicative of a cemetery origin. Location of discovery, condition of the remains, and the types of associated artifacts are all factors that aid forensic anthropologists in identifying cemetery remains.     

Recognition of cemetery remains in a forensic context

The purpose of this paper is to provide guidelines for the recognition of human remains from modem and historic cemeteries found in a forensic context. Three avenues of evidence may be pursued to confirm the origin of cemetery remains: context, condition of the body, and associated artifacts. This article outlines types of North American cemeteries, demonstrating how land use over time has resulted in many being closed, moved, or forgotten, leaving only the context to indicate their presence. The condition of human cemetery remains varies considerably depending on cultural practices and burial environment, but many exhibit combinations of the following traits: dried or embalmed tissue; erosion of bony pressure points; cortical bone flaking; and bone damage due to autopsy or embalming. Examples of artifact types useful in recognizing cemetery remains are also provided. Two cases from British Columbia, Canada are presented to demonstrate the diagnostic features of a disturbed cemetery burial.     

Recognition of cemetery remains in the forensic setting

Cemetery remains exposed through vandalism or natural phenomena are frequently brought to the attention of law enforcement agents or medical examiners. Although it is often difficult to distinguish cemetery remains from those of medicolegal significance, clues to their origin may exist. Characteristics consistent with cemetery remains include physical characteristics associated with the embalming process. Characteristics indicative of cemetery remains include functional or ornamental artifacts associated with the coffin, devices used in embalming the body, and elevated levels of embalming chemicals in the soft tissue.     

Acute Aortic Dissection Diagnosed after Embalming: Macroscopic and Microscopic Findings

A 58‐year‐old man died suddenly in Madagascar and poisoning was suspected. The body was embalmed after death and the general state of preservation was good. We found a major aortic dissection with a large false lumen from the aortic root to the common iliac arteries and a hemopericardium with formalinized blood clot. The intimal tear was on the ascending aorta, and an intramural hemorrhage was noted at the right coronary artery, attesting to a retrograde dissection. Microscopic studies confirmed aortic dissection with extensive intramural hemorrhage and also confirmed the retrograde dissection to the right coronary artery with a reduction of 90% of the true lumen. Classically, aortic dissection occurs in individuals with hypertension and individuals with genetic disorders of collagen formation. The diagnosis is often first established at the postmortem examination. Aortic dissection is therefore dealt with largely in necropsy studies. The usual cause of death is rupture into the pericardial sac. One case of bloodless dissection has been reported but the sudden death was explained by acute myocardial ischemia secondary to dissection of the left coronary artery. In our case, we found major hemopericardium and also intramural hemorrhage at the right coronary artery. We were able to make the diagnosis of aortic dissection and exclude the suspicion of homicide 15 days after death and after embalming.     

Determination of ethchlorvynol in body tissues and fluids after embalmment

A 54 year-old female expired at her residence. Her husband, a physician, signed a certificate stating that her death was due to cerebrovascular accident (CVA) and released her body to a funeral home, where she was embalmed. Since the deceased had a long history of medical problems and drug abuse, an autopsy was performed and no evidence of CVA was found. Toxicological analyses of body fluids and tissues revealed the presence of ethchlorvynol in high concentration in the bile (112 mg/l). The bloody fluid collected from the heart contained a concentration of ethchlorvynol below the limit for quantitation. Other findings included phenobarbital (32.8 mg/l) in heart bloody fluid and methanol (an ingredient of embalming fluid). The significance of the findings is discussed in relation to embalmment prior to autopsy and toxicological analyses. Ethchlorvynol concentration in the bile is compared to other fatal cases due to ethchlorvynol overdose.     

The influence of storage temperature and chemical preservation on the stability of succinylcholine in canine tissue

Succinylcholine (SCh) has been detected six months postmortem in liver, kidney, and injection site muscle of rats given 10 to 200 mg/kg by intramuscular injection. SCh stability was studied in canine tissue to evaluate three storage temperatures and two chemical preservatives at three time periods after injection. Nine mongrel dogs weighing 17.2 to 28 kg were divided equally into three groups and administered either 0.5, 1.0, or 5.0 mg SCh/kg intravenously into the cephalic vein. Liver, kidney, and gastrocnemius muscle were removed 90 min post-injection and divided into twelve portions. Each portion was treated with embalming fluid, physostigmine, the combination (50/50), or nothing. Chemically treated tissues and nontreated tissues were then stored at either 27, 5, or -20 degrees C for a period of up to forty days. Tissue portions were analyzed using ion-pair extraction, chemical demethylation, and gas chromatography with nitrogen phosphorous detection. Stability of SCh was greatest for samples stored at -20 degrees C and preserved with the combination of embalming fluid plus physostigmine. Kidney concentrations of SCh were significantly higher than those in liver or muscle at all doses. SCh was detected 24 h post-injection in all cases. By 40 days, only trace amounts of SCh, if any, could be detected in samples stored at room temperature with no chemical preservatives.     

Postmortem tissue embolisms. Report of 3 cases

In three cases of autopsy (cause of death - intoxication and drowning) we found in the lung intraalveolar und intravasal lymphatic tissue and bone-marrow. The results cannot be explained with violence during lifetime, but in tow cases as an effect of fracture of rips caused by external cardiacmassage, in one case by embalming with injection of conservation fluids.     

Determination of heroin after embalmment

A 30-year-old male died in Thailand after a scuffle. The corpse was embalmed and repatriated to France where an autopsy was performed. As usual in cases of embalmment, fluids such as blood and urine were unavailable and the toxicological analyses was performed on the bile and the liver. An overdose of heroin was determined as the cause of death. A review of the literature indicates that several drugs can be detected in fluids and tissues that contain formaldehyde. This case demonstrates that in embalmed corpses, toxicological assessment is still possible, e.g. after heroin fatalities.     

Evaluation of a Freezer Mill for Bone Pulverization prior to DNA Extraction: An Improved Workflow for STR Analysis

Traditional methods for bone pulverization typically generate heat, risking stability of DNA sample. SPEX? has developed cryogenic grinders which introduce liquid nitrogen to cool the sample and aid in the grinding process. In this study, the Freezer Mill 6970 EFM was used with two DNA extraction methods and routine downstream STR analysis procedures. DNA from as little as 0.1 g of bone powder was used to develop full STR profiles after freezer mill pulverization, and the method was reproducible. Further, no contamination was detected upon cleaning/reuse of the sample vials. There were no significant differences in DNA yield, STR alleles detected, or peak heights using the freezer mill as compared to traditional grinding, and successful DNA profiles were achieved from as low as 0.1 g of bone powder with this method. Overall, this work indicates that this cryogenic mill method may be used as a viable alternative to traditional tissue grinders.     

THE IMPACT OF FLORIDA'S HABITUAL OFFENDER LAW ON CRIME*

Research examining the impact of habitual offender laws on crime has suffered from numerous methodological problems, including failure to consider incapacitation effects that may be responsible for most of the laws' impact but that may not appear until years after the law is passed; not addressing simultaneity issues; and omitting needed control variables to avoid spurious or suppressed results. The present study specifies procedures that can be used to mitigate these problems. These procedures are used in the current study to estimate the impact of Florida's habitual offender law on crime using a multiple time series design, with data for 58 counties from 1980–1998.     

Development and Validation of a Technique for Casting Anatomical Features and Toolmarks

Time may elapse between examination of marks inflicted on the body and comparison with a potential weapon. Two‐dimensional photographs may be inadequate for effective comparison of a three‐dimensional mark with a putative instrument. Taking a cast and producing a positive image results in a lasting three‐dimensional record. This project aimed to develop and demonstrate the accuracy of an easy technique for production of long‐lasting positive images (using plaster of Paris and dental alginate). Casts of facial features of embalmed cadavers were used to produce a positive image of the face (death mask). Measurements of distances between facial anatomical landmarks were compared with measurements of distances between the same landmarks on the death masks to assess accuracy of reproduction. There were no significant differences between cadaver and death mask in 6 of 9 measurements, indicating this technique has high accuracy with less mobile facial features, but produces deformation of pliable features.     

Diagnosis of carbon monoxide poisoning following embalming and exhumation

Clots of blood, which were found in the cardiac ventricles at an autopsy of an embalmed corpse, were to analyse for carbon monoxide. It was not possible to determine reliably the CO-Hb-content of the blood clots with the known methods. Therefore a new procedure was developed. Carbon monoxide was released by a solution of nitric acid and determined gas chromatographically by a headspace method. In the same vessel the iron-content of the sample was measured after decomposition with atomic absorption spectrometry. This procedure is especially suitable for samples like putrid or heat-clotted blood.     

The effectiveness of various strategies to improve DNA analysis of formaldehyde-damaged tissues from embalmed cadavers for human identification purposes

Formalin-fixed tissues provide the medical and forensic communities with alternative and often last resort sources of DNA for identification or diagnostic purposes. The DNA in these samples can be highly degraded and chemically damaged, making downstream genotyping using short tandem repeats (STRs) challenging. Therefore, the use of alternative genetic markers, methods that pre-amplify the low amount of good quality DNA present, or methods that repair the damaged DNA template may provide more probative genetic information. This study investigated whether whole genome amplification (WGA) and DNA repair could improve STR typing of formaldehyde-damaged (FD) tissues from embalmed cadavers. Additionally, comparative genotyping success using bi-allelic markers, including INDELs and SNPs, was explored. Calculated random match probabilities (RMPs) using traditional STRs, INDEL markers, and two next generation sequencing (NGS) panels were compared across all samples. Overall, results showed that neither WGA nor DNA repair substantially improved STR success rates from formalin-fixed tissue samples. However, when DNA from FD samples was genotyped using INDEL and SNP-based panels, the RMP of each sample was markedly lower than the RMPs calculated from partial STR profiles. Therefore, the results of this study suggest that rather than attempting to improve the quantity and quality of severely damaged and degraded DNA prior to STR typing, a more productive approach may be to target smaller amplicons to provide more discriminatory DNA identifications. Furthermore, an NGS panel with less loci may yield better results when examining FD samples, due to more optimized chemistries that result in greater allelic balance and amplicon coverage.     

Biotechnology and law: biotechnology patents. Special considerations on the inventions with human material.

The EC and the US differ considerably in their ruling on the patentability of biotechnological inventions apart from some exceptions. The most extreme positions in either direction with regard to the individual biotechnological inventions mark the margins of the scope which the WTO members may use for the ratification of Art. 27 TRIPS. Rulings which are not in accordance with Art. 27 TRIPS remain without consideration. In the most important point both WTO member agree: Human beings are not patentable. Other regulations could be considered, but one may assume that no member of the WTO intends to make use thereof. Thus the scope involving this option is only a theoretical one. The situation is different when it comes to the components of the human body. These may, provided they are produced artificially or are separated from the human body, be patented. According to the regulations in the US, this applies to all components with the exception of totipotent stem cells. They may be exempted from patenting entirely or their patenting is linked to far reaching conditions. As an example should be named here the regulation of the EPA for patenting gene sequences. The scope of this area is, thus, very great and allows for many options of applying the regulations of Art. 27 TRIPS. With regard to patenting procedures for cloning human beings, the scope is unlimited. These procedures may either be completely excluded from patenting as in the EC or may be considered patentable independent of their purpose as in the US. The same applies to procedures for gathering human stem cells in the US. Exempting procedures with regard to human genes or DNA sequences are, however, only possible in special cases, as for example in the EC with regard to procedures for altering the genetic identity of the germline of human beings. However, the interpretation of general patentability preconditions allows to influence the patenting of such procedures. This means, though, that the scope for interpretation is only a small one in this area.     

蛋白质降解与死亡时间推断的初步研究

目的观察大鼠肝脏肌动蛋白、微管蛋白在死后不同时间的降解情况,为死亡时间的推断寻找客观依据。方法大鼠麻醉致死后置于21℃温度控制系统模拟死后18d的改变,在不同时间点剪取肝脏组织抽提蛋白质,利用westernblot技术检测肌动蛋白、微管蛋白的降解变化并对产物进行半定量分析。结果大鼠肝脏肌动蛋白在死后8d尚可检测,死亡10d后检测不到;在死后2d,α微管蛋白已检测不到,但可检测到β微管蛋白,死亡4d后,β微管蛋白也检测不到。结论肌动蛋白、微管蛋白的死后降解存在差异,其在肝脏组织保存时间的不同可作为死亡时间推断的参数。     

The relationship of methanol and formate concentrations in fatalities where methanol is detected

An automated headspace gas chromatography method was developed for the determination of formate (formic acid) in postmortem specimens, based on the in situ sulfuric acid-methanol methylation of formic acid to methyl formate. Diisopropyl ether was used as an internal standard. The method was applied to over 150 postmortem cases where methanol was detected. Of the 153 cases presented, 107 deaths were attributed to acute methanol toxicity. In the vast majority of the remaining 46 deaths, the methanol was determined to be present as a postmortem or perimortem artifact, or was otherwise incidental to the cause of death. Of the 76 victims who were found dead and blood was collected by the medical examiner, all but one had a postmortem blood formate concentration greater than 0.50 g/L (mean 0.85 g/L; n = 74). The sole exception involved suicidal ingestion of methanol where the blood methanol concentration was 7.9 g/L (790 mg/100 mL) and blood formate 0.12 g/L. In 97% (72/74) of the cases where blood was available, the blood formate was between 0.60 and 1.40 g/L. In 31 of the 153 cases, the victim was hospitalized and blood obtained on admission or soon after was analyzed for methanol and formate during the subsequent death investigation; the vast majority (27/30) had antemortem blood formate concentrations greater than 0.50 g/L. Cases with samples taken prior to death with blood formate concentrations less than 0.5 g/L can readily be explained by active treatment such as dialysis. The blood formate method has also been useful in confirming probable perimortem or postmortem contamination of one of more fluids or tissues with methanol (e.g., windshield washer fluid or embalming fluid), where methanol ingestion was unlikely.     

Fatal trauma to the neck: immunohistochemical study of local injuries

As fatal trauma to the neck is often associated with short survival times, proof of vitality may often be difficult using standard histochemical techniques. Soft tissue neck injuries resulting from strangulation by ligature or manual strangulation were examined immunohistochemically using antibodies to myoglobin, fibronectin, C5b-9 and MRP14, and compared to controls consisting of accidental soft tissue neck injuries as well as undamaged neck soft tissue. Although survival times in the study and control groups were unknown and certainly some individual variation may be expected in the time course of normal wound development, both the study and control groups demonstrated similar time courses in the immunohistochemical detection of antigen. Myoglobin was always found in those samples in which only one antigen was shown to be involved in an injury-specific pattern; myoglobin and fibronectin were found in samples with dual antigen involvement. Samples involving three antigens always included C5b-9 in addition to myoglobin and fibronectin. The single positive MRP14 sample in the study and control groups was simultaneously positive for the other markers used. Myoglobin, fibronectin, C5b-9 and MRP14 are therefore suitable for immunohistochemical detection of vital reactions and estimation of temporal relationships in the early posttraumatic period after neck trauma.     

A Fatality Following Ingestion of the Designer Drug Meta‐Chlorophenylpiperazine (mCPP) in an Asthmatic—HPLC‐MS/MS Detection in Biofluids and Hair

A 20‐year‐old man, a cocaine addict and regular ecstasy user, with a medical history of allergic asthma died after ingesting half a tablet earlier the same day. The white tablet, stamped with a “smiling sun” logo looked very much like an ecstasy tablet and was sold as such. He experienced a severe asthma attack just after ingesting the half tablet and it evolved over the next few hours into fatal cardiorespiratory arrest. Biological samples, taken after embalming, were analyzed by high‐performance liquid chromatography tandem mass spectrometry (HPLC‐MS/MS). Analysis revealed meta‐chlorophenylpiperazine (mCPP) in concentrations of 45.8 mg in a similar tablet obtained later from the drug dealer, 5.1 ng/mL in the bile, 0.3 ng/g in the liver, 15.0 ng/mL in the urine, and its absence in a hair sample (<0.02 ng/mg), which indicated he was not a regular user (whereas strong concentrations of MDMA and cocaine were found in the hair). Interrogated by the police after his arrest, the dealer said that he had sold the victim and for the very first time two tablets with the same “smiling sun” logo. The tablet used for analysis was from the same brand as the one ingested by the victim. The autopsy excluded other causes of death, while the histological analyses showed a large number of polynuclear eosinophils in the bronchial walls, confirming the asthmatic pathology. None of the other organs examined (larynx, liver, heart, adrenal glands, and kidneys) showed any distinctive signs, and in particular no inflammatory infiltrate. The death was the result of an asthma attack in an asthmatic person, violently decompensated following ingestion of approximately 20 mg of mCPP.