Fatal Overdose of Gamma‐hydroxybutyrate Acid After Ingestion of 1,4‐Butanediol

We report a case of fatal intoxication from 1,4‐butanediol (1,4‐BD), which was ingested by a young and “naïve” gamma‐hydroxybutyrate (GHB) consumer during a party with the co‐ingestion of alcohol, cannabis, and methylene‐dioxy‐methamphetamine. The following drug concentrations were found using gas chromatography coupled with mass spectrometry on autopsy samples and on a cup and a glass found at the scene: 20,350 mg/L (bottle) for 1,4‐BD; 1020 mg/L (femoral blood), 3380 mg/L (cardiac blood), 47,280 mg/L (gastric content), and 570 mg/L (vitreous humor) for GHB. The concentration of GHB is difficult to interpret in forensic cases due to the possibility of an endogenous production of GHB. The variable tolerance of the user may also modify the peri‐ and postmortem GHB concentrations. This case underscores the need to have many different sources of toxicology samples analyzed to avoid the hypothesis of endogenous production of GHB.     

An Evidence‐Based Forensic Taxonomy of Windows Phone Communication Apps

Communication apps can be an important source of evidence in a forensic investigation (e.g., in the investigation of a drug trafficking or terrorism case where the communications apps were used by the accused persons during the transactions or planning activities). This study presents the first evidence‐based forensic taxonomy of Windows Phone communication apps, using an existing two‐dimensional Android forensic taxonomy as a baseline. Specifically, 30 Windows Phone communication apps, including Instant Messaging (IM) and Voice over IP (VoIP) apps, are examined. Artifacts extracted using physical acquisition are analyzed, and seven digital evidence objects of forensic interest are identified, namely: Call Log, Chats, Contacts, Locations, Installed Applications, SMSs and User Accounts. Findings from this study would help to facilitate timely and effective forensic investigations involving Windows Phone communication apps.     

Fatty Acid Profiles for Differentiating Growth Medium Formulations Used to Culture Bacillus cereus T‐strain Spores

Microbial biomarkers that indicate aspects of an organism's growth conditions are important targets of forensic research. In this study, we examined fatty acid composition as a signature for the types of complex nutrients in the culturing medium. Bacillus cereus T‐strain spores were grown in medium formulations supplemented with one of the following: peptone (meat protein), tryptone (casein protein), soy protein, and brain–heart infusion. Cellular biomass was profiled with fatty acid methyl ester (FAME) analysis. Results showed peptone cultures produced spores enriched in straight‐chained lipids. Tryptone cultures produced spores enriched in branched‐odd lipids when compared with peptone, soy, and brain–heart formulations. The observed FAME variation was used to construct a set of discriminant functions that could help identify the nutrients in a culturing recipe for an unknown spore sample. Blinded classification tests were most successful for spores grown on media containing peptone and tryptone, showing 88% and 100% correct identification, respectively.     

“YFlag” – A Single‐base Extension Primer Based Method for Gender Determination

Assigning the gender of a DNA contributor in forensic analysis is typically achieved using the amelogenin test. Occasionally, this test produces false‐positive results due to deletions occurring on the Y chromosome. Here, a four‐marker “YFlag” method is presented to infer gender using single‐base extension primers to flag the presence (or absence) of Y‐chromosome DNA within a sample to supplement forensic STR profiling. This method offers built‐in redundancy, with a single marker being sufficient to detect the presence of male DNA. In a study using 30 male and 30 female individuals, detection of male DNA was achieved with c. 0.03 ng of male DNA. All four markers were present in male/female mixture samples despite the presence of excessive female DNA. In summary, the YFlag system offers a method that is reproducible, specific, and sensitive, making it suitable for forensic use to detect male DNA.     

Evaluation of Stature Estimation from the Database for Forensic Anthropology*†

Abstract: Trotter and Gleser’s ( 1 - 3 ) stature equations, conventionally used to estimate stature, are not appropriate to use in the modern forensic context. In this study, stature is assessed with a modern (birth years after 1944) American sample (N = 242) derived from the National Institute of Justice Database for Forensic Anthropology in the United States and the Forensic Anthropology Databank. New stature formulae have been calculated using forensic stature (FSTAT) and a combined dataset of forensic, cadaver, and measured statures referred to as Any Stature (ASTAT). The new FSTAT‐based equations had an improved accuracy in Blacks with little improvement over Ousley’s ( 4 ) equations for Whites. ASTAT‐based equations performed equal to those of FSTAT equations and may be more appropriate, because they reflect both the variation in reported statures and in cadaver statures. It is essential to use not only equations based on forensic statures, but also equations based on modern samples.     

Anion Identification via Complexation with meso‐octamethylcalix(4)pyrrole and Detection Using Electrospray Ionization Mass Spectrometry*

Abstract: The routine identification of controlled substances and adulterants during forensic chemistry analysis often involves the identification of counter ions or salt forms present in an exhibit. Here, the use of the compound meso‐octamethylcalix(4)pyrrole (C4P) during salt‐form identification analysis is presented. C4P is a commercially‐available, anion‐binding agent that can be reacted with a controlled substance or adulterant, resulting in the sequestration of anionic species, usually present as counter ions to the active ingredient. Formation of noncovalent complexes between the cyclic host C4P compound and anionic guests is investigated using electrospray ionization–mass spectrometry (ESI–MS). Complexes with chloride, bromide, iodide, nitrate, and acetate are readily observed and mass spectrometry analysis provides identification via molecular weight characterization. Chloride and bromide complexes are also characterized by the isotopic distribution of their molecular ions. Formation of host–guest complexes is not observed for sulfate and phosphate salts, presumably due to steric hindrance and energetically unfavorable conditions.     

Dermestes maculatus (Coleoptera: Dermestidae) development under fluoxetine effect using two drug administration models

Fluoxetine is a selective serotonin reuptake inhibitor, commonly used for the treatment of a variety of psychopathological conditions. As such, fluoxetine may be expected to appear in clinical and forensic cases. Dermestes maculatus De Geer (Coleoptera: Dermestidae) has been recognized as a relevant component of the insect fauna associated with decomposing human and animal remains. Experiments were conducted to study the effect of fluoxetine on developing D. maculatus using two‐drug administration models: a non‐living animal model (pork muscle) and a living one (Sus scrofa L. pigs). We assessed the duration of immature stages and total life cycle, as well as morphological parameters (body length, cephalic width, and weight). The effect of fluoxetine was studied at an overdose concentration: In the non‐living animal model the drug was mixed with macerated pork muscle (2000 mg/kg) and in the living animal model, pigs were given the drug orally (833 mg/kg). A control was used for each model. Daily observations were performed from the beginning to the end of the experiments. GC‐MS was used for drug detection and quantification. There were no statistically significant differences in the duration of immature stages, life cycle, larval mortality, morphological parameters, or sex ratio, between treatment and control, regardless of the drug administration model. Given that fluoxetine had no detectable effect on the development of D. maculatus, detection of this drug in forensic situations would not compromise the accuracy of PMI estimations.     

The Effects of Body Mass on Cremation Weight*

Abstract: Cremains have become increasingly frequent in forensic contexts, while higher body mass in the general population has simultaneously made cremation a more cost‐effective mortuary practice. This study analyzed the relationship between body mass and bone mass, as reflected through cremation weight. Antemortem data were recorded for samples used in the multi‐regional data set. Each was rendered through commercial crematoriums and reweighed postincineration. Pearson’s correlation demonstrates clear association between body mass and cremation weight (r = 0.56; p < 0.0001). However, multiple linear regression revealed sex and age variables also have a significant relationship (t = 7.198; t = ?2.5, respectively). Regressed in conjunction, body mass, sex, and age contribute approximately 67% of all variation observed in cremation weight (r = 0.668). Analysis of covariance indicates significant regional variation in body and cremation weight. Explanations include bone modification resulting from increased loading stress, as well as glucose intolerance and altered metabolic pathways related to obesity.     

Development of a Tetraplex PCR Assay for CYP2D6 Genotyping in Degraded DNA Samples

CYP2D6 polymorphism analysis is gaining increasing interest in forensic pharmacogenetics. Nevertheless, DNA recovered from forensic samples could be of poor quality and not suitable for long polymerase chain reaction required to type CYP2D6 gene prior to SNaPshot minisequencing analysis performed to define alleles with different enzymatic activity. We developed and validated following the guidelines of the Scientific Working Group on DNA Analysis Methods a tetraplex PCR yielding four amplicons of 597, 803, 1142, and 1659 bp encompassing the entire CYP2D6 gene to analyze eleven SNP positions by SNaPshot minisequencing. Concordance, sensitivity, and specificity were assessed. The method, applied to thirty‐two forensic samples failed to amplify with long PCR, allowed the amplification of CYP2D6 gene in 62.5% of degraded samples. The new tetraplex PCR appears a suitable method for CYP2D6 analysis in forensic pharmacogenetics.     

Measurement of Postmortem Pupil Size: A New Method with Excellent Reliability and Its Application to Pupil Changes in the Early Postmortem Period

Measurement of postmortem pupil width is a potential component of death time estimation. However, no standardized measurement method has been described. We analyzed a total of 71 digital images for pupil–iris ratio using the software ImageJ. Images were analyzed three times by four different examiners. In addition, serial images from 10 cases were taken between 2 and 50 h postmortem to detect spontaneous pupil changes. Intra‐ and inter‐rater reliability of the method was excellent (ICC > 0.95). The method is observer independent and yields consistent results, and images can be digitally stored and re‐evaluated. The method seems highly eligible for forensic and scientific purposes. While statistical analysis of spontaneous pupil changes revealed a significant polynomial of quartic degree for postmortem time (p = 0.001), an obvious pattern was not detected. These results do not indicate suitability of spontaneous pupil changes for forensic death time estimation, as formerly suggested.     

Whole Plastome Sequences of Two Drug-Type Cannabis: Insights Into the Use of Plastid in Forensic Analyses

DNA is one of the fastest growing tools in forensic sciences, increasing reliability in forensic reports and judgments. The use of DNA has increased in different areas of the forensic sciences, such as investigation of plant species, where plastid DNA has been used to elucidate and generate evidence in cases of traceability of genetically modified and controlled plants. Even with several advances and the practice of using DNA in forensic investigations, there are just few studies related to the identification of genetic tools for the characterization of drug and nondrug-types of Cannabis. Herein, the whole plastomes of two drug-type Cannabis are presented and have their structures compared with other Cannabis plastomes deposited in the GenBank, focusing in the forensic use of plastome sequences. The plastomes of Cannabis sativa “Brazuka” and of the hybrid Cannabis AK Royal Automatic presented general structure that does not differs from the reported for other C. sativa cultivars. A phylogenomic analyses grouped C. sativa “Brazuka” with the nondrug C. sativa cultivars, while the hybrid Cannabis AK Royal Automatic placed isolated, basal to this group. This suggests that the analysis of plastomes is useful toward genetic identification of hybrids in relation to C. sativa.     

Validation of thin layer chromatography with AccuTOF-DART™ detection for forensic drug analysis

Abstract: Thin layer chromatography (TLC) is a technique that is commonly employed in the forensic drug analysis of pharmaceutical preparations. Detection is typically accomplished using various visualization spray reagents. Conventional gas chromatography–mass spectrometry (GC‐MS) analysis is typically performed to confirm the TLC results. Depending on the drugs tested and the instrument conditions required, this confirmation can take up to an hour to complete. Direct analysis in real time (DART?) is an ionization source, coupled to an accurate‐mass time‐of‐flight mass spectrometer that has the capability to ionize materials under ambient conditions. To streamline analysis, the combination of TLC with DART? detection is proposed to screen and subsequently identify drug compounds, all from the same TLC plate. DART? confirmations of TLC analyses take <10 min to complete and compare favorably to GC‐MS in sensitivity and selectivity. This study validates the use of TLC‐DART in the forensic identification of the components of several pharmaceutical preparations.     

The Odontology Victim Identification Skill Assessment System*,†

Abstract: Mass fatality identification efforts involving forensic odontology can involve hundreds of dental volunteers. A literature review was conducted and forensic odontologists and dental educators consulted to identify lessons learned from past mass fatality identification efforts. As a result, the authors propose a skill assessment system, the Odontology Victim Identification Skill Assessment System (OVID‐SAS), which details qualifications required to participate on the Antemortem, Postmortem, Ante/Postmortem Comparison, Field, and Shift Leader/Initial Response Teams. For each qualification, specific skills have been identified along with suggested educational pedagogy and skill assessment methods. Courses and assessments can be developed by dental schools, professional associations, or forensic organizations to teach and test for the skills required for dental volunteers to participate on each team. By implementing a system, such as OVID‐SAS, forensic odontologists responsible for organizing and managing a forensic odontology mass fatality identification effort will be able to optimally utilize individuals presenting with proven skills.     

A Three‐Locus,PCR‐based Method for Forensic Identification of Plant Material

Plant residue is currently an underutilized resource in forensic investigations despite the fact that many crime scenes, as well as suspects and victims, harbor plant‐derived residue that could be recovered and analyzed. Notwithstanding the considerable skill of forensic botanists, current methods of species determination could benefit from tools for DNA‐based species identification. However, DNA barcoding in plants has been hampered by sequence complications in the plant genome. Following a database search for usable barcodes, broad‐spectrum primers were designed and utilized to amplify and sequence the rbcL, trnL‐F, and rrn18 genetic loci from a variety of household plants. Once obtained, these DNA sequences were used to design species‐targeted primers that could successfully discriminate the source of plant residue from among the 21 species tested.     

Measurement of Postmortem 1,5‐anhydroglucitol in Vitreous Humor for Forensic Diagnosis

In forensic diagnosis, postmortem blood glucose is known to be susceptible to change after death. However, the 1,5‐anhydroglucitol (1,5‐AG) concentrations in plasma and cerebrospinal fluid (CSF) reflect the mean blood glucose level for a short period of time. In this study, we compared the postmortem 1,5‐AG concentrations in vitreous humor and CSF in 47 subjects to evaluate the utility of this concentration in the vitreous humor for forensic diagnosis. The postmortem 1,5‐AG concentrations in vitreous humor (mean±SD: 20.2 ± 8.7 μg/mL) and CSF (16.8 ± 8.7 μg/mL) did not differ significantly and showed a strong correlation (r² = 0.87, p < 0.01). These results suggest that the vitreous humor 1,5‐AG concentration provides useful information on the antemortem blood glucose level, in addition to the HbA1c value and the CSF 1,5‐AG concentration.     

A Polydrug Intoxication Involving Methoxetamine in a Drugs and Driving Case

Methoxetamine ((RS)2‐(3‐methoxyphenyl)‐2‐(ethylamino)cyclohexanone)) is becoming a drug of interest among practitioners of forensic toxicology. In this case report, we describe the case background, standard field sobriety tests, sampling, and analysis of this drug in a whole blood sample as well as screening methods and analysis from a driver operating under the influence of intoxicating substances. Methoxetamine was isolated from the blood sample using mixed mode solid phase extraction. After elution and evaporation, the residue was dissolved in mobile phase (consisting of acetonitrile and aqueous formic acid) for analysis by liquid chromatography–tandem mass spectrometry (LC–MS/MS) and gas chromatography–mass spectrometry (GC–MS). The case sample was found to contain clonazepam, 7‐aminoclonazepam, carboxy‐THC, Ddphenhydramine, and MDMA. The case sample was found to contain 10 ng/mL of the drug (methoxetamine) in whole blood. The results of this drug analysis and previous analyses are discussed in terms of this driver operating under the influence of drugs.     

Testing a Novel 3D Printed Radiographic Imaging Device for Use in Forensic Odontology

There are specific challenges related to forensic dental radiology and difficulties in aligning X‐ray equipment to teeth of interest. Researchers used 3D printing to create a new device, the combined holding and aiming device (CHAD), to address the positioning limitations of current dental X‐ray devices. Participants (N = 24) used the CHAD, soft dental wax, and a modified external aiming device (MEAD) to determine device preference, radiographer's efficiency, and technique errors. Each participant exposed six X‐rays per device for a total of 432 X‐rays scored. A significant difference was found at the 0.05 level between the three devices (p = 0.0015), with the MEAD having the least amount of total errors and soft dental wax taking the least amount of time. Total errors were highest when participants used soft dental wax—both the MEAD and the CHAD performed best overall. Further research in forensic dental radiology and use of holding devices is needed.     

A Test of the Passalacqua Age at Death Estimation Method Using the Sacrum

A test of the accuracy of the Passalacqua (J Forensic Sci, 5, 2009, 255) sacrum method in a forensic context was performed on a sample of 153 individuals from the J.C.B. Grant Skeletal Collection. The Passalacqua (J Forensic Sci, 5, 2009, 255) method assesses seven traits of the sacrum using a 7‐digit coding system. An accuracy of 97.3% was achieved using the Passalacqua (J Forensic Sci, 5, 2009, 255) method to estimate adult skeletal age. On average each age estimate differed by 12.87 years from the known age. The method underestimated the age of individuals by an average of 4.3 years. An intra‐observer error of 6.6% suggests that the method can be performed with precision. Correlation and regression analysis found that the sacral traits used in the Passalacqua (J Forensic Sci, 5, 2009, 255) method did not have a strong relationship with age or an ability to strongly predict age. Overall, the method was not practical for use in a forensic context due to the broad age ranges, despite the high accuracy and low intra‐observer error.     

DNA Barcoding Identifies all Immature Life Stages of a Forensically Important Flesh Fly (Diptera: Sarcophagidae)

Carrion‐breeding insects, such as flesh flies (Diptera: Sarcophagidae), can be used as evidence in forensic investigations. Despite their considerable forensic potential, their use has been limited because morphological species identification, at any life stage, is very challenging. This study investigated whether DNA could be extracted and cytochrome oxidase subunit I (COI) barcode sequences obtained for molecular identification of each immature life stage of the forensically important Australian flesh fly, Sarcophaga (Sarcorohdendorfia) impatiens (Walker). Genomic DNA extracts were prepared from all larval instars and puparia. Amplification of the barcoding region was successful from all extracts, but puparia amplicons were weak. All sequences were identified as S. impatiens with 99.95% confidence using the Barcoding of Life Database (BOLD). Importantly, crop removal was necessary to eliminate PCR inhibition for specimens from late second and early third instars. Similar results are expected for immatures of other carrion‐breeding species, enhancing the use of evidence from immature flies in forensic investigations.