Entomotoxicology.

Entomotoxicology is a relatively new branch of forensic entomology. The potential use of insects for detecting drugs and other toxins in decomposing tissues has been widely demonstrated. In death investigations, Diptera and other arthropods can be reliable alternate specimens for toxicological analyses in the absence of tissues and fluids normally taken for such purposes. Entomotoxicology also investigates the effects caused by drugs and toxins on arthropod development in order to assist the forensic postmortem interval estimates. However, several remarks on the limitations of entomotoxicology have been highlighted recently. In this paper, the implications for the practice of this forensic procedure are fully reviewed.     

法医昆虫毒理学在药(毒)物检测中的应用与发展

作为重要的毒物检测新途径,法医昆虫毒理学在推断高度腐败或白骨化尸体生前染毒状况方面已愈发显示出其独特优势.本文概括了法医昆虫毒理学在药(毒)物检测领域的发展历程,整理了检测所涉及各药(毒)物、相应阳性昆虫枪材,阐述了通过昆虫样本进行毒理学检测的优势,并对尸-虫间药物浓度相关性进行了分析总结.     

Morphine extraction in necrophagous insects remains for determining ante-mortem opiate intoxication.

To study the potential use of insects remains as toxicological indicators, we measured morphine in desiccated entomological materials using a radioimmunoassay method (Coat-A-Count Serum Morphine, Dade Behring, France) following enzymatic hydrolysis.First, enzymatic cuticle extraction of morphine (pronase digestion preceded by a 2h incubation with a dithiothreitol solution) was performed on various substrates (Calliphoridae puparial cases and desiccated adults, and desiccated pupae of Dermestidae) which were then tested to identify possible interferences with the radioimmunoassay procedure.This same procedure was performed on puparial cases and desiccated adults of Lucilia sericata (Diptera, Calliphoridae) previously reared on minced meat containing various morphine concentrations. Morphine was detected only in cuticle powder from insects reared on meats containing 100 and 1000mg/kg. Higher concentrations were measured in puparial cases. Rearings on psoas from eight heroin overdose victims confirmed previous experimental results.Remains of necrophagous insects, particularly puparial cases, are often preserved for a long time, and consequently can serve as late alternative specimens for toxicological analysis when suitable tissues are not available.     

Toxicological Analysis of Opiates from Alternative Matrices Collected from an Exhumed Body

In this case study, the body of a 45‐year‐old man was exhumed after 1 year at the request of the public prosecutor to assess whether the death was caused by drug consumption. Toxicological analyses were performed on several matrices, including liver, kidney, and the alternative matrices hair and teeth. The systematic toxicological analysis (STA), which consisted of basic and acid liquid/liquid extraction and gas chromatography–mass spectrometry (GC‐MS) analysis, showed the presence of opiates in each of the matrices analyzed. Subsequently, to confirm and quantify the presence of opioids, samples of each of the matrices were subjected to solid‐phase extraction and specific GC‐MS analysis. The case presented demonstrates the possibility of drug detection in an exhumed body that has been buried for 1 year, despite the problems of quantitative interpretation of the data, and that toxicological results could be useful along with other forensic evidence.     

Analysis of the Effect of Cyclophosphamide and Methotrexate on Chrysomya megacephala (Diptera: Calliphoridae),

Forensic entomotoxicology investigates the effects of chemical substances in the development of scavenger insects and the reflection on estimating the minimum postmortem interval (PMI). To evaluate the impact of the chemotherapeutic drugs cyclophosphamide (CF) and methotrexate (MTX) on the postembryonic development of Chrysomya megacephala, controlled experiments were performed by rearing the larvae on spiked minced beef with different concentrations of these drugs. The results indicated that CF significantly decreased their developmental rate up to 28 h but had no significant effect on larval and adult sizes, survival rate, and sex ratio, whereas MTX decreased larval and adult sizes, survival rate, and there was a deviation in the expected sex ratio toward females in MTX‐exposed larvae but had no significant impact on developmental rate. These negative interference factors should be considered in cases of suspected death of people that have undergone chemotherapy.     

Prodrugs of New Psychoactive Substances (NPS): A New Challenge

The concept of a substance acting as a prodrug for an intended drug is not new and has been known and utilized with particular benefits within medicine for efficacy and patient safety. Prodrugs of psychoactive substances are also not particularly new but this has also extended to considerations of prodrugs of new psychoactive substances (NPS). The continuing evolution of NPS has been a constant forensic challenge. In some countries, this constant evolution has led to the introduction of various alternative methods of drug control. Whether for this reason or in the pursuit of user experimentation, prodrugs of NPS have been discussed, developed, and exploited, posing some distinct forensic challenges. This is especially the case within toxicological analysis of biological fluids and for some substances, also forensic chemical analysis, through inherent instability of the prodrug or metabolism in the body. Particular examples of NPS prodrugs include 1-propanoyl-LSD, 1-butanoyl-LSD, 1-acetyl-LSD, and 2C-B-AN. This is in addition to associated substances and medicines that may be used for an intended pharmacological effect. Various prodrugs for stimulant and hallucinogenic substances in particular have appeared in the literature and have been discussed within drug user forums and made available for purchase online. Presently, drug monitoring data from national and international systems indicate that prodrugs are not widely available or problematic. Nevertheless, it is important that there is sufficient awareness of the prodrug concept and potential impact and associated forensic implications, not just for chemical analysis but also for toxicological considerations when a substance has been used.     

Love,fear, and justice: Transforming selves for the new world

Two current models of the self are contrasted. The limitations of these (egocentric autonomous and sociocentric organic) models generate five problems for a comprehensive approach to justice. An alternative model is presented that views the self as an agent who is dependent on others and is motivated by both love for the other and fear for the self. This alternative model has three advantages: The two current models can be derived from it; it resolves the problems created by their limitations; it provides a concrete vision of how we must develop ourselves if we wish to obtain a just world.     

The cocaine "body stuffer" syndrome: a fatal case

Body stuffer, sometimes called "mini packer", is the definition of someone who admits to or is strongly suspected of ingesting illegal drugs in order to escape detection by authorities, and not for recreational purposes or to transport the drug across borders. Cocaine is the drug most commonly involved in the body stuffer syndrome. Reported cases of body stuffer deaths are rare, however a fatality related to the ingestion of a plastic bag containing cocaine is described regarding a 17-year-old dealer. The authors describe how the cocaine body stuffer syndrome differs from the usual body packer. Histological and toxicological findings are examined and discussed for a better definition of this unique syndrome.     

Metabolism and toxicological detection of the designer drug N-(1-phenylcyclohexyl)-3-methoxypropanamine (PCMPA) in rat urine using gas chromatography-mass spectrometry

Studies on the metabolism and the toxicological detection of the phencyclidine-derived designer drug N-(1-phenylcyclohexyl)-3-methoxypropanamine (PCMPA) in rat urine are described using gas chromatographic-mass spectrometric (GC-MS) techniques. Based on the identified metabolites, the following metabolic pathways could be postulated: N-dealkylation, O-demethylation partially followed by oxidation of the resulting alcohol to the corresponding carboxylic acid, hydroxylation of the cyclohexyl ring at different positions, and aromatic hydroxylation. The formed metabolites were identical to those of the homologue N-(1-phenylcyclohexyl)-3-ethoxypropanamine (PCEPA) with exception of the mono hydroxyl metabolites of PCEPA. All PCMPA metabolites were partially excreted in conjugated form. An intake of a common drug users' dose of PCMPA could be detected in rat urine by the authors' systematic toxicological analysis (STA) procedure using full-scan GC-MS after acid hydrolysis, liquid-liquid extraction and microwave-assisted acetylation. The STA should be suitable for proof of an intake of PCMPA also in human urine assuming similar metabolism.     

Standardization of Histological Procedures for the Detection of Toxic Substances by Immunohistochemistry in Dipteran Larvae of Forensic Importance

Immunohistochemistry (IHC) technique is an alternative toxicological analysis to detect drugs in insects of forensic importance, but it requires thorough histological procedures. In this study, we tested different fixatives—phosphate‐buffered paraformaldehyde 4% (PP), Carnoy's fluid (CF), Kahle's solution (KS), ethanol in different concentrations, and ethanol associated to PP and CF, time of fixation and histological processes for dipteran larvae's tissue, aiming to develop a sample preparation protocol for IHC application. A suitable fixation was achieved using PP for 12 and 24 h, CF for 3 h, 70% ethanol for 19 days, and 70% ethanol/CF for 2 h/3 h. Postfixation using negative pressure, two immersions in xylene for 30 min each, and one in xylene plus paraffin for 45 min increased tissue preservation. An immunohistochemical test for cocaine detection was performed using monoclonal benzoylecgonine antibody from mouse, peroxidase‐conjugated anti‐mouse IgG and visualized by 3,3′‐diaminobenzidine method showed these histological procedures didn't compromise antigenicity.     

Fatal intoxication involving 2-methyl AP-237

Novel synthetic opioids contribute considerably to the opioid epidemic, especially with the frequent emergence of structurally similar compounds. This case report describes a fatal intoxication involving 2-methyl AP-237. A 54-year-old Caucasian male was found deceased from an apparent drug overdose. A plastic container labeled “2MAP” and a cut straw were found in the decedent's backpack at the scene. A white substance found in the container tested positive for fentanyl by field testing. According to his medical history, the decedent was treated for a drug overdose 3 years prior to his death. With no diagnostic findings at autopsy, the case was submitted for toxicological analysis. An unknown substance was detected in peripheral blood and urine using gas chromatography with nitrogen phosphorous detection (GC-NPD). Further testing was conducted using gas chromatography–mass spectrometry (GC–MS) and liquid chromatography-quadrupole-time-of-flight mass spectrometry (LC-QTOF-MS) which confirmed the presence of 2-methyl AP-237 and potential metabolites in blood and urine. Quantitation by GC-NPD revealed concentrations of 2-methyl AP-237 in blood and urine at 480 ng/mL and 4200 ng/mL, respectively. The toxicological analysis also identified and quantitated alprazolam in the blood at 55 ng/mL. Additionally, the metabolism of 2-methyl AP-237 was investigated and three hydroxylated metabolites were identified in peripheral blood and urine. Limited literature is available for the detection and quantitation of 2-methyl AP-237 in postmortem specimens. Given the toxicological findings with unremarkable autopsy findings, this case is an example of a fatal intoxication involving 2-methyl AP-237.     

Application of forward‐looking infrared (FLIR) imaging from an unmanned aerial platform in the search for decomposing remains

Traditional methods of cadaver detection in outdoor environments include manual ground search, cadaver dogs, and manned aerial reconnaissance during daylight. These methods have limitations; however, a potential low‐cost alternative may be to employ thermal imaging equipment mounted on an unmanned aerial vehicle (UAV) to detect heat emitted by insect and bacterial activity on the decomposing remains. No studies have addressed the influence of wrappings on detection of maggot mass thermal signatures nor assessed thermal detection of smaller body fragments. We addressed these knowledge gaps by utilizing a two‐phase experimental approach to explore thermal detection of carcasses using UAV‐mounted infrared imaging. In Phase 1, pig body fragments were deposited on the surface or shallow buried. In Phase 2, whole pig carcasses were deposited in four conditions: on the surface uncovered, wrapped in plastic or carpet, or buried. Our results demonstrated that observable heat emissions from remains corresponded to peak insect activity during active decay and could be readily detected in uncovered whole carcasses and fragments. Although plastic and carpet wrappings partly impeded detection of insect heat signatures, these materials were clearly detectable themselves because of their contrast to the background ground surface. Thermal signatures of buried partial remains and disturbed gravesoil were also observed; however, the buried whole carcass transitioned to adipocere prior to the decay stage and without any insect colonization or heat signature. These data can inform operational implementation of this technique to complement existing search strategies to offer a robust, low‐cost alternative for use where scene characteristics allow.     

Toxicology and sudden infant death

One hundred thirty cases of sudden infant death occurring in Wayne County, Michigan, (population 2.7 million) were analyzed for possible drugs. The toxicological protocol has been outlined. Six cases were found to be positive, and in five of these the drugs found had been prescribed for a variety of illnesses. In one case methadone was found in the blood of an infant whose mother was undergoing methadone treatment for drug addiction, the drug being transmitted through breast milk. In none of the 102 cases of sudden infant death syndrome (SIDS) included in the study did the toxicological results affect the diagnosis. In our study, toxicological analyses never contradicted an initial diagnosis of SIDS, and, therefore, we feel that this diagnosis should be made promptly based on investigative and autopsy findings.     

mtDNA多态性在嗜尸性昆虫种类鉴定中的应用

嗜尸性昆虫的种类鉴定是利用昆虫学知识进行法医学研究的重要步骤之一.近年来研究表明,利用分子生物学技术,特别是线粒体DNA(mitochondrial DNA,mtDNA)序列分析技术对嗜尸性昆虫进行种类鉴别,能够得到较好的效果.本文对嗜尸性昆虫mtDNA的分子生物学特性、结构以及嗜尸性昆虫种类鉴定中常用的mtDNA序列...     

Fatal intoxication with melperone

Melperone is judged to be a safe neuroleptic drug. Until now there has been no report of a melperone fatality, though it has been used in suicide attempts. We report on a case of a 36-year-old woman where no cause of death could be established at autopsy. Criminological investigation pointed to a homicide by poisoning but also the possibility of a suicide had to be taken in account. The toxicological analysis of blood, cerebral spinal fluid and urine revealed extremely high concentrations of melperone which had never been reported before. Furthermore, diazepam, nordazepam and carbamazepine were detected. To our knowledge, this case is the first melperone fatality. Possible interactions with diazepam and carbamazepine are discussed.     

Alternative sanctions for drug users: fruitless efforts or miracle solution?

In most Western European countries, including Belgium, judicial alternative sanctions are increasingly being used for drug users. Because no study into the effectiveness of Belgian judicial alternatives for drug users has yet been carried out, this became the objective of the current research. The design of this study comprises a pre and post measurement of the criminal activity, drug use and situation in different spheres of life of 565 drug-dependent offenders. Two conclusions can be drawn. First, after an alternative sanction or measure is imposed, there is a reduction in the criminal activity of the offender. Second, this crime reduction goes hand in hand with a progress in several relevant life spheres.     

Hair analysis of opiates in mothers and newborns for evaluating opiate exposure during pregnancy

The increasing interest in toxicological hair analysis as a marker of human exposure to xenobiotics such as illicit substances or therapeutic drugs, has been made feasible by the extension of mass spectrometry, a highly sensitive method of detection. A newborn exposed to drugs in utero can suffer from a varying degree of withdrawal syndrome, a few days after birth. If of opiate origin, the withdrawal syndrome can be treated with morphine, among other therapeutics, but it is not easy to diagnose because of atypical symptoms presented by neonates and especially when maternal drug addiction has not been revealed. To assess and measure toxicological factors linked with the appearance and the severity of this syndrome, maternal and neonatal matrices such as urine, meconium and hair were collected during a protocol approved by the ethical committee. Opiates in particular were measured with GC-MS and potential combined dependences (cannabis, cocaine, amphetamine, LSD and benzodiazepines) and/or substitutive therapeutics (methadone or buprenorphine) were also assessed in 17 mother/neonate couples. Gestational opiate exposure profiles were drawn up and linked with the observed withdrawal syndromes. A withdrawal syndrome seems to appear more frequently after foetal exposure to an association of opiates/substitutive molecules (8 out of 10 withdrawal syndromes observed in this study), although the impact of cocaine and benzodiazepines must also be taken into account. The results obtained in neonatal hair make it possible to affirm foetal drug exposure and are in accordance, for the majority, with the appearance of a neonatal withdrawal syndrome (NWS). Neonatal hair analysis could contribute to assess in utero exposure to opiates, particularly when results in urine and meconium are negative or when these matrices are not available.     

Differentiation between drug use and environmental contamination when testing for drugs in hair

The differentiation between systemic exposure and external contamination for certain drug groups has been frequently referred to as one of the limitations of in drug testing in hair. When hair samples are used, three steps are usually employed in order to minimise the possibility of external contamination causing a misinterpretation. The first consists of decontaminating hair samples by washing the hair before analysis, the second is the detection of the relevant metabolites in the hair samples and the third is the use of cut-off levels. Difficulty in the interpretation arises when metabolites are not detected either due to external contamination of the hair or low doses of the drugs used. A wash protocol needs to be practical and ideally remove any drug deposited on the external portion of the hair.