An unsuccessful murder

The victim of an attempted murder, who was shot in the head by her husband, survived when the bullet failed to penetrate her skull, apparently because he had scored the bullet twice across its nose and used an outdated power charge. The husband succeeded in killing himself with a second shot.     

A study of distribution of ethyl glucuronide in different keratin matrices

Ethyl glucuronide (EtG) is a direct metabolite of ethanol, frequently used as a biomarker of alcohol abuse. To this purpose, EtG is preferentially determined in hair samples, using a cut-off value of 30pg/mg to discriminate between social and heavy drinkers, as recently fixed by an international consensus conference. Although this cut-off value is assumed for head hair, alternative matrices, such as pubic, axillary and chest hair, are often analyzed when head hair is not available. Previous studies suggested that determination of EtG in various keratin matrices may lead to different results; growth cycle and rate, urine contamination, distribution of sebum glands and other environmental factors are likely to contribute to these differences. We analyzed more than 2700 samples (head, pubic, chest and axillary hair) to evaluate the inter- and intra-individual distribution of the EtG concentration in the different keratin matrices. The data were interpreted on a statistical basis, on the assumption that large population data-sets will level off the average alcohol consumption of each group. From both inter- and intra-individual distribution data, significant differences were observed in EtG concentrations recorded in head, axillary and pubic hair samples. It is concluded that pubic hair cannot be utilized alternatively to head hair to prove chronic alcohol abuse, nor is axillary hair, since positive and negative biases respectively affect these determinations. In contrast, for chest hair, EtG distributions similar to head hair were found, although the large discrepancy between the examined population dimensions presently prevents any definitive conclusion. Thus, chest hair represents a promising alternative to head hair for EtG determinations, deserving further investigation on samples collected from the same individuals, in order to establish a clear correlation between their respective EtG concentrations.     

Hemorrhage of the head and neck in death by burning

The 202 deaths connected with burns in 1976-1986 included 134 in which death occurred during the fire. In 49 cases the face was so badly charred that it was not possible to judge whether hemorrhage had occurred in the eyelids and/or conjuctivae. Such blood extravasation was seen in 14 of the remaining 85 cases, taking the form of petechiae in some cases and in others the form associated with more extensive extravasation, in some cases hemolytic. No other features known to cause such hemorrhages were present, so that they are attributed to the burning process itself. Signs of extensive pronounced burns were found in 13 of these cases, and it seemed that high-degree burns on the neck and trunk with less severe burns on the head were particularly likely to be associated with such hemorrhages. In most cases the flames had quickly come into direct contact with the body (clothing, bed or chair had caught fire), leading to rapid death, as documented by the fact that usually very little or no soot at all had been aspirated and by the low level of carbon monoxide intoxication (max. COHb 22%). One plausible explanation for the development of hemorrhage is the supposition first expressed over 20 years ago that when the circulation is maintained rapid burn-induced shrinkage of the skin of the neck has the effect of strangulating the victim; hemorrhage of eyelids and/or conjunctivae in such cases be could regarded as a vital reaction to the effect of fire. This is particularly significant, insofar as evidence of hemorrhage of this kind was found predominantly in bodies in which other vital signs indicative of the effects of burning were sparse and slight or even totally absent. In addition, specific examination of the laryngeal area revealed congestion-induced extravasation at various points, as well as petechial hemorrhage in the mucous membrane.     

Ethyl glucuronide: unusual distribution between head hair and pubic hair

Ethyl glucuronide (EtG) is a minor metabolite of ethanol that can be detected in hair. In some specific situations, head hair can be missing, and therefore, alternative anatomical locations of hair are of interest. In this study, paired hair specimens (head hair and pubic hair) from eight social drinkers were analyzed for EtG. Each sample was decontaminated by two dichloromethane bathes (5 ml) for 2 min. After cutting into small pieces, about 50 mg of hair was incubated in 2 ml water in the presence of 10 ng of EtG-d5, used as internal standard and submitted to ultra-sonication for 2 h. The aqueous phase was extracted by SPE using Oasis MAX columns. The hair extract was separated on an ACQUITY BEH HILIC column using a gradient of acetonitrile and formate buffer. Detection was based on two daughter ions: transitions m/z 221-85 and 75 and m/z 226-75 for EtG and the IS, respectively. This laboratory is using a positive cut-off at 50 pg/mg. All eight head hair specimens were negative for EtG at a limit of quantitation fixed at 10 pg/mg. Surprisingly, EtG was identified at high concentrations in pubic hair, in the range 12-1370 pg/mg. It appears, therefore, that it is not possible to document the drinking status of a subject by simply switching from head hair to pubic hair.     

A comparative study on the concentrations of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (THCCOOH) in head and pubic hair

In this study, the concentrations of 11-nor-Δ(9)-tetrahydrocannabinol-9-carboxylic acid (THCCOOH) in pubic, axillary and beard hair were measured and the correlation between the concentrations of THCCOOH in head and pubic hair from same cannabis users were evaluated. The papers on body hair analysis for THCCOOH were rarely found although police officers submit body hair as a complimentary specimen to forensic laboratories in case cannabis users had no hair. Head, pubic, axillary, and beard hair were collected. All hair samples were cut into 0.5mm segments and decontaminated with methanol, digested with 1 mL of 1M NaOH at 85 °C for 30 min and extracted in 2 mL of n-hexane:ethyl acetate (9:1) two times after adding 1 mL of 0.1N sodium acetate buffer (pH = 4.5) and 200 μL of acetic acid followed by derivatization with 50 μL of PFPA and 25 μL of PFPOH for 30 min at 70 °C. The extracts were analyzed using gas chromatography tandem mass spectrometry operating in negative chemical ionization mode (GC/MS/MS-NCI). We determined the concentrations of THCCOOH in both pubic and head hair. The concentrations of THCCOOH in pubic hair were higher than those in head hair. We also evaluated the concentrations of THCCOOH in body hair (pubic, axillary and beard hair) and head hair according to the positive/negative urine test results. There was no statistically significant difference in the concentrations of THCCOOH in head and body hair according to urine results.     

Detection of thiopental and pentobarbital in head and pubic hair in a case of drug-facilitated sexual assault

The quali-quantitative determination of two barbiturates, thiopental and its metabolite pentobarbital, in head and pubic hair samples of a woman who had been sexually assaulted during hospitalisation, is reported. Hair was analysed by means of solid-phase microextraction (SPME) and gas chromatography-multiple mass spectrometry (GC-MS-MS), in chemical ionisation conditions. Thiopental and pentobarbital were found in three proximal head hair segments (sample 1A: 0.30 and 0.40 ng/mg; sample 1B: 0.20 and 0.20 ng/mg; sample 3: 0.15 and 0.20 ng/mg) and pubic hair sample. Two distal head hair segments were negative for both barbiturates. Despite the lack of collection and toxicological analysis of blood or urine samples within the hospital setting, analytical findings from hair revealed the use of the anaesthetic agent thiopental to sedate the victim quickly and deeply and commit sexual assault.     

Methadone concentrations in human hair of the head, axillary and pubic hair

The concentrations of methadone and its metabolites in the hair of the head, axillary and pubic hair obtained from patients receiving a daily maintenance doses, were determined. Comparison of the concentrations provides the highest values in the axillary hair, followed by pubic hair and the hair of the head.     

Hair as a specimen to document tetramethylene disulfotetramine exposure

Tetramethylene disulfotetramine (tetramine) is a rodenticide that has been banned for many years in China. Since 2005, inhabitants of a village in the Henan Province have been suffering from grand mal seizures. To investigate the possibility of tetramine as the cause, we developed a method to determine tetramine in human hair. Sample preparation involved external decontamination, frozen pulverization, and ultrasonication in 2 mL ethyl acetate in the presence of cocaine-d3 as an internal standard. The method exhibited good linearity; calibration curve was linear over a range of 0.1-20 ng/mg hair. The limit of detection for the assay was 0.05 ng/mg hair. Except for one subject (No. 4), all head and pubic hair samples were positive for tetramine. The concentrations of tetramine in pubic hair were significantly higher than those in the same subjects' head hair samples. Because of a long retention in body, segmental head hair analysis cannot provide an accurate exposure history of tetramine in the body.     

Arsenic poisoning: acute or chronic? Suicide or murder?

The case of the death by arsenic poisoning of a 62-year-old white man is presented. One year prior to death, he developed intermittent bouts of severe gastroenteritis with vomiting and diarrhea, hyperpigmentation and keratosis of the skin, neutropenia, and Guillain-Barré-like neuropathy for which he was hospitalized several times. Urine test results 6 months prior to death indicating 36 mg/L arsenic were believed to be in error. At the patient's last admission, he appeared in the emergency room with severe gastroenteritis, hypotension, and dehydration. He died 3 days later. Antemortem as well as autopsy specimens revealed elevated arsenic concentrations. Arsenic micrograms/g analysis by neutron activation of hair pulled from the man's head revealed by centimeter segmental analysis proximal to distal: 226, 104, 28, 56, 41, 40, and 74. The wife of the decedent was charged with murder by arsenic poisoning of this, her fifth, husband. The defense alleged that the decedent had committed suicide. The judge awarded a directed verdict of "not guilty." Particulars of the medical, toxicological, and investigative findings are presented.     

Drowning due to cyclobenzaprine and ethanol.

The deceased was a 35 year old female who was found by her husband in the bathtub with her head and face submerged in the water. Autopsy findings were unremarkable. Toxicological analysis revealed the presence of ethanol (215 mg/dL) and cyclobenzaprine (1.786 mg/l) in the blood. This high concentration of cyclobenzaprine (Flexeril) in combination with alcohol proved to be fatal.     

Hair transfers in sexual assault: a six-year case study

Associative head and pubic hair transfers can provide a circumstantial connection between persons and objects in sexual assault cases. The occurrence of reported hair transfers in one analyst's casework are presented.     

Tetrahydrocannabinols in hair of hashish smokers

The study investigates the presence of tetrahydrocannabinols in the head hair and the pubic and axillary hair. The hair samples were obtained from hashish smokers. The concentrations were determined by radioimmunoassay and reflect total tetrahydrocannabinols and metabolites. The values found ranged from 0.4 ng/mg hair up to 3.8 ng/mg hair. The presence of the drug in the hair samples was also demonstrated by GC/MS.     

Correlation of methamphetamine results and concentrations between head, axillary, and pubic hair

This study was designed to compare the qualitative results and concentrations of methamphetamine (MA) and its metabolite amphetamine (AP) in head hair and hair collected from different parts of the body (axillae and pubis). Hair from subjects (N = 14) suspected MA users was simultaneously collected. Hair preparation involved washing step, fine cutting, overnight extraction, derivatization by the trifluoroacetic anhydride, and gas chromatography/mass spectrometry (GC/MS) using selective ion monitoring. In this study, we found a good correlation of the qualitative results for MA between head hair and hair on other parts of the body, but there were some differences in concentrations of MA and AP. Namely, the concentrations of MA and AP were higher in axillary and pubic hair than in head hair.     

Distribution of 6-monoacetylmorphine and morphine in head and pubic hair from heroin-related deaths

There is limited published data to aid interpretation of analytical findings from hair analysis. The aim of the study was to collate 6-monoacetylmorphine (6-AM) and morphine concentrations in head and pubic hair from heroin users and to propose reference ranges and relate these to the amount of heroin used. The ratio of morphine-to-6-AM was also investigated. A total of 82 head hair samples divided into 173 segments of various lengths and 15 pubic hair samples were collected at postmortem from heroin users. The hair was analysed using a previously published method. A statistical evaluation demonstrated that in head hair, the lower, middle and upper concentration ranges of 6-AM were 0.1–0.9, 0.9–12.5 and 12.5–154.1 ng/mg and those of morphine were 0.1–0.8, 0.8–6.0 and 6.0–36.3 ng/mg. In pubic hair, the lower, middle and upper concentration ranges of 6-AM were 0.2–0.5, 0.5–2.3 and 2.3–18.2 ng/mg and those of morphine were 0.2–0.4, 0.4–2.4 and 2.4–13.3 ng/mg. The morphine-to-6-AM ratio showed a large variation. The ratio tended to decrease from proximal to distal segments. The statistical results suggest low, middle and high concentration ranges which we propose can be used for estimating the amount of heroin consumed into corresponding low or occasional, regular or habitual and heavy or excessive drug use. The ratio of morphine-to-6-AM showed great variation and did not support the proposal that a ratio less than 0.77 is needed to prove ingestion of heroin.     

Sometimes What Everybody Thinks They Know Is True

This essay responds to D. Davis and W. C. Follette (2002), who question the value of motive evidence in murder cases. They argue that the evidence that a husband had extramarital affairs, that he heavily insured his wife's life, or that he battered his wife is ordinarily of infinitesimal probative value. We disagree. To be sure, it would be foolish to predict solely on the basis of such evidence that a husband will murder his wife. However, when this kind of evidence is combined with other evidence in a realistic murder case, the evidence can be quite probative. We analyze cases in which it is virtually certain that the victim was murdered but unclear who murdered her, and in which it is uncertain whether the husband murdered the wife or she died by accident. We show that in each case motive evidence, such as a history of battering or of infidelity, can substantially increase the odds of the husband's guilt. We also consider the actual case on which Davis and Follette base their paper. We argue that testimony of Davis on the basis of the analysis presented in their paper was properly excluded, for it would have been misleading and unhelpful.     

Death by lightning

Deaths from lightning injuries are infrequent, amounting to 0.2-0.8 per million people per year, and are associated with climatologic conditions. The victims are mostly young, active people who are struck during various outdoor activities during the summer months. Even though many people can be struck by the same bolt of lightning, multiple casualties are uncommon. Among the fatalities, various types of skin burns are regularly present, often at an entry site on or near the head, and at an inferiorly located exit site. These skin injuries often consist of superficial burns and singed hair; deep burns and charring are uncommon. Arborescent skin marks seem to occur less frequently. Internal findings at autopsy are nonspecific.     

Histopathological differentiation of skin lesions caused by electrocution, flame burns and abrasion

The aim of this study was to reveal histopathological features for differential diagnosis of skin lesions caused by electrocution, flames and abrasions. Based on the causes of the lesions, cases were assigned into three groups. Group 1 included 30 deaths from electric shock. Group 2 included 30 individuals with flame burns who died in the fires. Group 3 included 30 deaths from traffic accidents, from which the individuals had abrasions. Data from the crime scene investigations and macroscopic and microscopic findings from the autopsies allowed determination of the cause of death in all cases. The features of the lesions examined under the light microscope were intraepidermal separation, subepidermal (dermoepidermal) separation, coagulation necrosis in the epidermis, nuclear elongation in the epidermis, dark-staining epidermal nucleus, depth of homogenization in the dermis, and nuclear elongation in the epithelium of hair follicles. A significantly high rate of electrical lesions had intraepidermal separation. The rate of subepidermal separation was slightly more significant in flame burns. A significantly higher rate of electrical lesions had both intraepidermal and subepidermal separation. The rate of coagulation necrosis in the epidermis was significantly the highest in electrical lesions. Although the severity of nuclear elongation was the most significant in electrical lesions, varying degrees of nuclear elongation in the epidermis were present in all three groups. Dark staining of the epidermal nuclei was present in all lesions except for one electrical lesion, though the severity of staining was significant in the abrasion group. The depth of homogenization was slightly more significant in the abrasion group. The rate of nuclear elongation in the epithelium of the hair follicles was significantly lower in the abrasion group. The results of this study revealed that certain morphological changes determined under a light microscope could help the differential diagnoses of electrical lesions, flame burns and abrasions.     

Results of hair analyses for drugs of abuse and comparison with self-reports and urine tests

Urine as well as head and pubic hair samples from drug abusers were analysed for opiates, cocaine and its metabolites, amphetamines, methadone and cannabinoids. Urine immunoassay results and the results of hair tests by means of gas chromatography-mass spectrometry were compared to the self-reported data of the patients in an interview protocol. With regard to the study group, opiate abuse was claimed from the majority in self-reports (89%), followed by cannabinoids (55%), cocaine (38%), and methadone (32%). Except for opiates the comparison between self-reported drug use and urinalysis at admission showed a low correlation. In contrast to urinalysis, hair tests revealed consumption in more cases. There was also a good agreement between self-reports of patients taking part in an official methadone maintenance program and urine test results concerning methadone. However, hair test results demonstrated that methadone abuse in general was under-reported by people who did not participate in a substitution program. Comparing self-reports and the results of hair analyses drug use was dramatically under-reported, especially cocaine. Cocaine hair tests appeared to be highly sensitive and specific in identifying past cocaine use even in settings of negative urine tests. In contrast to cocaine, hair lacks sensitivity as a detection agent for cannabinoids and a proof of cannabis use by means of hair analysis should include the sensitive detection of the metabolite THC carboxylic acid in the lower picogram range.     

Immunohistochemical analysis of forensic evidence from a double homicide

We report the use of immunohistochemical staining for analysis of forensic evidence from a double homicide. A 38-year-old woman and her 7-year-old daughter were murdered by multiple blows to the head and face with a tomahawk, resulting in multiple fragments of brain tissue scattered about the murder scene. The victims' husband and father was the main suspect, who maintained that he was out of town on business during the evening of the murders. However, a shirt taken from the suspect's car on the morning after the murders (secured by the police before the suspect visited the murder scene) was found to have two small stains. DNA analysis on the stains showed the presence of the deceased wife's DNA, and immunohistochemical stains on shirt fragments conclusively documented the presence of deep central nervous system tissue, providing the critical piece of evidence needed to arrest and prosecute the suspect. This report demonstrates that shirt or similar cloth fragments can be processed into paraffin blocks and subsequently immunostained to search for and classify types of tissue fragments that may be present on the fabric.