Journal of Forensic Sciences Available Online 17 May 2013

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Butalbital and Driving Impairment

by Jillian K. Yeakel, Barry K. Logan

Abstract

Butalbital (Fiorinal^®), used in the treatment of migraines and muscle pain, is the most commonly encountered barbiturate in impaired driving cases. It has central nervous system (CNS) depressant properties, including sedation, drowsiness, and feelings of intoxication, which can contribute to driving impairment. Twenty-six driving under the influence cases are reviewed including results from field sobriety tests and toxicology testing. Blood samples were screened using enzyme multiplied immunoassay technique immunoassay, and the presence of butalbital was confirmed and quantified using gas chromatography/mass spectrometry, gas chromatography with flame ionization detection, or gas chromatography nitrogen/phosphorus detection. Butalbital concentrations ranged from 1.0 to 30.2 mg/L, with a mean and median of 16.0 mg/L. General impairment indicators in these cases included horizontal and vertical nystagmus, lack of convergence, poor motor coordination, and balance and speech problems, which are common to CNS depressant intoxication, similar to that associated with alcohol. These findings indicate the importance of toxicological testing for butalbital in cases where CNS depressants are indicated.

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

by Carina M. Souza, Carolina G. P. Lima, Marcos J. Alves-Jr, Wagner W. Arrais-Silva, Selma Giorgio, Arício X. Linhares, Patricia J. Thyssen

Abstract

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.

Direct Analyte-Probed Nanoextraction Coupled to Nanospray Ionization–Mass Spectrometry of Drug Residues from Latent Fingerprints

by Kristina Clemons, Rachel Wiley, Kristin Waverka, James Fox, Eric Dziekonski, Guido F. Verbeck

Abstract

Here, we present a method of extracting drug residues from fingerprints via Direct Analyte-Probed Nanoextraction coupled to nanospray ionization–mass spectrometry (DAPNe-NSI-MS). This instrumental technique provides higher selectivity and lower detection limits over current methods, greatly reducing sample preparation, and does not compromise the integrity of latent fingerprints. This coupled to Raman microscopy is an advantageous supplement for location and identification of trace particles. DAPNe uses a nanomanipulator for extraction and differing microscopies for localization of chemicals of interest. A capillary tip with solvent of choice is placed in a nanopositioner. The surface to be analyzed is placed under a microscope, and a particle of interest is located. Using a pressure injector, the solvent is injected onto the surface where it dissolves the analyte, and then extracted back into the capillary tip. The solution is then directly analyzed via NSI-MS. Analyses of caffeine, cocaine, crystal methamphetamine, and ecstasy have been performed successfully.

Objective Color Classification of Ecstasy Tablets by Hyperspectral Imaging

by Gerda Edelman, Martin Lopatka, Maurice Aalders

Abstract

The general procedure followed in the examination of ecstasy tablets for profiling purposes includes a color description, which depends highly on the observers’ perception. This study aims to provide objective quantitative color information using visible hyperspectral imaging. Both self-manufactured and illicit tablets, created with different amounts of known colorants were analyzed. We derived reflectance spectra from hyperspectral images of these tablets, and successfully determined the most likely colorant used in the production of all self-manufactured tablets and four of five illicit tablets studied. Upon classification, the concentration of the colorant was estimated using a photon propagation model and a single reference measurement of a tablet of known concentration. The estimated concentrations showed a high correlation with the actual values (R² = 0.9374). The achieved color information, combined with other physical and chemical characteristics, can provide a powerful tool for the comparison of tablet seizures, which may reveal their origin.

High-Throughput STR Analysis for DNA Database Using Direct PCR,

by Jeong Eun Sim, Su Jeong Park, Han Chul Lee, Se-Yong Kim, Jong Yeol Kim, Seung Hwan Lee

Abstract

Since the Korean criminal DNA database was launched in 2010, we have focused on establishing an automated DNA database profiling system that analyzes short tandem repeat loci in a high-throughput and cost-effective manner. We established a DNA database profiling system without DNA purification using a direct PCR buffer system. The quality of direct PCR procedures was compared with that of conventional PCR system under their respective optimized conditions. The results revealed not only perfect concordance but also an excellent PCR success rate, good electropherogram quality, and an optimal intra/inter-loci peak height ratio. In particular, the proportion of DNA extraction required due to direct PCR failure could be minimized to <3%. In conclusion, the newly developed direct PCR system can be adopted for automated DNA database profiling systems to replace or supplement conventional PCR system in a time- and cost-saving manner.

Sudden Unexpected Death due to Hemophagocytic Lymphohistiocytosis Syndrome

by Dennis J. Chute, James Rawley, John Cox, Robert J. Bready

Abstract

The hemophagocytic lymphohistiocytosis (HLH) syndrome is a hyperimmune disorder characterized by lymphohistiocytic infiltrations, elevated cytokine levels in the blood, macrophage activation, and hemophagocytosis, frequently presenting with a febrile septic picture. This unusual disease is more common in infancy and childhood than adulthood. It is classified as primary or familial when a genetic defect is identified and secondary or acquired when triggered by certain infections, autoimmune disorders, or malignancies. If or when such patients expire, they typically do so within a hospital or under a physician’s care and so such cases rarely come to the attention of forensic pathologists. We report on the unexpected deaths of two hospitalized adult cases of HLH brought to autopsy without a premortem diagnosis. Postmortem examination demonstrated marked hepatosplenomegaly and lymphadenopathy in association with hemophagocytosis. Although very uncommon HLH must be considered in infants, children, or adults who die unexpectedly with an undiagnosed septic presentation.

A Novel Application of Infrared Imaging for the Diagnosis of Death by Fire

by Shinobu Yamauchi, Masahito Hitosugi, Hajime Kimura, Shogo Tokudome

Abstract

This paper demonstrates a novel and simple technique for the diagnosis of death by fire using a digital infrared (IR) camera system. At autopsy, the inhalation of soot by a fire victim is a definite indicator proving a vital reaction during the fire. However, there can be difficulties in confirming the presence of soot because of the relative lack of soot or the similarity in color between soot and surrounding tissues. To evaluate the effectiveness of IR imaging at autopsy, we acquired internal images of the respiratory and gastrointestinal system by both an ordinary color and IR cameras. Using our method, the inhaled soot is visible as black particles or deposits, while the blood is transparent and the surrounding tissue is whitened. This technique enables the detection of normally undiscriminating soot in an ordinary color image. This is the first report suggesting the usefulness of IR imaging in forensic autopsy for the diagnosis of death by fire.

A Comparison of DNA Extraction Using AutoMate Express™ and EZ1 Advanced XL from Liquid Blood, Bloodstains, and Semen Stains

by Koji Fujii, Shota Inokuchi, Tetsushi Kitayama, Hiroaki Nakahara, Natsuko Mizuno, Kazumasa Sekiguchi

Abstract

In this study, DNA was extracted using an AutoMate Express™ and an EZ1 Advanced XL from liquid blood, fresh and aged bloodstains, and fresh and aged semen stains. Extracted DNA was quantified by real-time PCR using the D17Z1 locus. Short tandem repeat typing was performed using an AmpFℓSTR^® Identifiler kit. The yields of DNA obtained by the AutoMate Express™ were higher from fresh bloodstains and fresh semen stains, almost the same from aged bloodstains and aged semen stains, but slightly lower from liquid blood compared with those obtained by the EZ1 Advanced XL. The addition of dithiothreitol or the use of PrepFiler™ lysis buffer improved the EZ1 Advanced XL results from fresh bloodstains, but not for liquid blood and aged bloodstains. Our results demonstrated that the PrepFiler™ lysis buffer is the main contributor to the higher DNA yields of the AutoMate Express™ for fresh bloodstains.

Subarachnoid Hemorrhage and Carbon Monoxide Exposure: Accidental Association or Fatal Link?

by Fabio De-Giorgio, Vincenzo M. Grassi, Massimo Miscusi, Cesare Mancuso, Ernesto d’Aloja, Vincenzo L. Pascali

Abstract

This case study reports a fatal subarachnoid hemorrhage (SAH) with concomitant accidental carbon monoxide (CO) exposure in 32-year-old man. Autopsy results indicated an antemortem aspiration of smoke, and a massive SAH was identified as the cause of death. Intriguingly, the carboxyhemoglobin level was 30%, suggesting that CO could have played a specific role. Intracranial hemorrhages following CO exposure in brain areas and tissues such as the basal ganglia, globus pallidus, or white matter are rare, but well characterized, whereas SAH related to CO exposure has not been previously described. In this case report, the possible role of CO, either as a primary cause or as a facilitating factor, in the pathogenesis of SAH is discussed. In particular, we propose the hypothesis that the excessive vasodilating effects produced by CO on the cerebral endothelium results in consequential loss of microvascular integrity.

Simultaneous Analysis of Micro-RNA and DNA for Determining the Body Fluid Origin of DNA Profiles,

by Donny Meer, Mari L. Uchimoto, Graham Williams

Abstract

Micro-RNAs (miRNAs) can be specifically expressed in forensically relevant body fluids such as blood or saliva. The aim of the study was to develop a simultaneous extraction and analysis protocol that allows for the acquisition of a DNA profile and the identity of the body fluid using a single process. DNA and micro-RNA were extracted from blood and saliva before undergoing a cDNA synthesis step by using stem-loop reverse transcription PCR. The resulting extracts containing DNA and cDNA synthesized from body fluid–specific miRNA markers then underwent standard STR analysis using a modified ABI AmpFℓSTR^® NGM SElect™ kit. In all samples, a full DNA profile was obtained along with additional peaks corresponding to the miRNA marker targeted. In all cases, blood samples profiled exhibited a peak indicating the presence of the blood-specific miRNA marker and the saliva sample profiled exhibited a peak indicating the presence of the saliva-specific miRNA marker.

Variability in the Blood/Breath Alcohol Ratio and Implications for Evidentiary Purposes

by Dena H. Jaffe, Maya Siman-Tov, Asher Gopher, Kobi Peleg

Abstract

The breath analyzer is an indispensable tool for identifying alcohol levels among drivers. While numerous studies have shown high correlations between blood and breath alcohol concentrations, most are limited by the study design. This study seeks to assess this relationship by minimizing potential measurement bias, document time from alcohol consumption to testing, and adjusting for potential confounders. A blinded study was performed using conditions closely resembling those in the field. The Draeger 7110 MKIII IL breath analyzer was used to assess breath alcohol concentrations (BrAC). Participants were 61 healthy volunteers aged 21–37 years with body mass index ≤30 and no history of alcoholism. A total of 242 valid blood/breath tests were performed in four test sets. The study results showed a high correlation coefficient between BrAC and blood alcohol concentration (BAC) levels (r = 0.983) with high sensitivity (97%) and specificity (93%). This strong association between the breath analyzer and BAC persisted even after adjustment for various stages of alcohol absorption. These results illustrate the high diagnostic sensitivity of the breath analyzer in field-tested conditions.

mtDNAprofiler: A Web Application for the Nomenclature and Comparison of Human Mitochondrial DNA Sequences,

by In Seok Yang, Hwan Young Lee, Woo Ick Yang, Kyoung-Jin Shin

Abstract

Mitochondrial DNA (mtDNA) is a valuable tool in the fields of forensic, population, and medical genetics. However, recording and comparing mtDNA control region or entire genome sequences would be difficult if researchers are not familiar with mtDNA nomenclature conventions. Therefore, mtDNAprofiler, a Web application, was designed for the analysis and comparison of mtDNA sequences in a string format or as a list of mtDNA single-nucleotide polymorphisms (mtSNPs). mtDNAprofiler which comprises four mtDNA sequence-analysis tools (mtDNA nomenclature, mtDNA assembly, mtSNP conversion, and mtSNP concordance-check) supports not only the accurate analysis of mtDNA sequences via an automated nomenclature function, but also consistent management of mtSNP data via direct comparison and validity-check functions. Since mtDNAprofiler consists of four tools that are associated with key steps of mtDNA sequence analysis, mtDNAprofiler will be helpful for researchers working with mtDNA. mtDNAprofiler is freely available at http://mtprofiler.yonsei.ac.kr.

40-Hz Square-Wave Stimulation Requires Less Energy to Produce Muscle Contraction: Compared with the TASER® X26 Conducted Energy Weapon

by James A. Comeaux, James R. Jauchem, D. Duane Cox, Carrie C. Crane, John A. D’Andrea

Abstract

Conducted energy weapons (CEWs) (including the Advanced TASER^® X26 model produced by TASER International, Inc.) incapacitate individuals by causing muscle contractions. In this study using anesthetized swine, the potential incapacitating effect of primarily monophasic, 19-Hz voltage imposed by the commercial CEW was compared with the effect of voltages imposed by a laboratory device that created 40-Hz square waves. Forces of muscle contraction were measured with the use of strain gauges. Stimulation with 40-Hz square waves required less pulse energy than stimulation with the commercial CEW to produce similar muscle contraction. The square-pulse stimulation, at the higher repetition rate, caused a more complete tetanus at a lower energy. Use of such a simple shape of waveform may be used to make future nonlethal weapon devices more efficient.

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Detection of diabetic metabolism disorders post-mortem – forensic case reports on cause of death hyperglycaemia

Drug Testing and Analysis 25 April 2013; DOI: 10.1002/dta.1479

Diabetic coma is the most severe form of hyperglycaemic metabolic disorders. The post-mortem diagnosis of this disorder of glucose metabolism can be difficult and vague due to a lack of characteristic morphological findings. Six death cases caused by diabetic coma are described with special focus on biochemical (and histological) findings. The possible glycaemia markers glucose, lactate, HbA1c, fructosamine, anhydroglucitol, and ketone bodies were measured and the usefulness of these parameters is evaluated and discussed. Estimations of glucose concentrations in vitreous humour or cerebrospinal fluid and of ketone bodies in blood or other matrices are obligatory while measurements of HbA1c, fructosamine, or anhydroglucitol can only provide additional information on the long-term adjustment of diabetes in the deceased. Lactate concentrations (addition of glucose and lactate levels to form the sum formula of Traub) do not give more information than the glucose concentration itself and can be therefore omitted. Copyright © 2013 John Wiley & Sons, Ltd.

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Evaluating the Use of DNA and RNA Degradation for Estimating the Post-Mortem Interval

United States National Institute of Justice.  NIJ Grant Number 2010-DN-BX-K228

Estimating the post-mortem interval (PMI) is difficult due to the many factors that influence the decomposition process. Tissues such as nails, teeth and bones are more resilient to environmental factors. Measuring the rate of degradation of nucleic acids in these tissues could provide a method for estimating the PMI for longer time intervals, from days and weeks, through to months and years. These time intervals have not been studied before in a systematic manner using human tissues. Our initial focus has been on nails. Nails are hard tissues, relatively easy to sample and until now, have been under-utilized as a tissue that has forensic value. We have investigated the stability of nucleic acids in nails to determine if both DNA and RNA can be co-extracted in levels suitable for PCR analysis. Furthermore, multiple nail samples can be taken from the same individual so that measuring changes in nucleic acid degradation can be undertaken where there are a minimal number of external variables so that the nucleic acid degradation is a more likely representation of the PMI.

Using different methods for extracting DNA and RNA from nails, we have found that by modifying the Promega DNA IQ™ method, both DNA and RNA are efficiently extracted. This modified method has been implemented into operational forensic casework at ESR and nails are now being utilized in more human identification cases.

We have developed multiplex PCR assays to measure the rate of degradation of messenger RNA (mRNA), ribosomal RNA (rRNA) and DNA. Using nails placed in different environmental conditions (air, soil and water), we have found that nails are protected from other environmental factors and that nucleic acids (both DNA and RNA) can be amplified from samples left submerged in water or placed in soil for 120 days (1043 accumulated degree days). Nails from human cadavers have been collected, and the DNA and RNA co- extracted. Using our multiplex PCR assays, we have shown that DNA and RNA can be co-extracted from cadaver nails with a PMI up to 20,925 ADDs.

Using statistical analyses, some of the nucleic acid amplified fragments (from DNA and mRNA) look promising to use for deriving a statistical model for estimating the PMI.

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Incaprettamento: an unusual homicide by ligature strangulation

Int. J. Criminol. Sociol. 2013; 2: 10-12;  http://dx.doi.org/10.6000/1929-4409.2013.02.2

This case study regards the murder of a Tunisian national residing in a small town in southern Italy who was murdered by fellow Tunisian citizen, also living in Italy. The victim was found hogtied and hanged by a cord that was suspended from an iron staff situated between two concrete electrical poles. This method of killing is known as “incaprettemento”. It is an unusual type of strangulation whereby a rope is passed around the victim’s wrists, ankles, and throat causing suffocation. This results in a very slow and painful death. Typically, it is a method employed by Italian organized crime groups, but it has also been used in the execution of war crimes. This means of killing is meant to impart a particularly degrading and humiliating revenge on the victim. It may also serve as sign of intimidation and a warning to others. There is very little written about incaprettamento in the literature, but it has, nonetheless, important psychological, anthropological, and cultural implications in the field of criminology. The perpetrator underwent psychodiagnostic evaluations, and was ultimately deemed fit to stand trial.

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Custody suicide with washrag: A case report with clinical and custodial consideration

Journal of Forensic and Legal Medicine  Volume 20, Issue 4, May 2013, Pages 343–344;  http://dx.doi.org/10.1016/j.jflm.2012.07.009

Custody or prison suicides with a washrag are rather unusual and few case reports present in the forensic literature. Completed suicide by using a washrag without the awareness of custody supervisors and medical staff ought to draw attention to related personnel. We report a case of suicide where the victim was a suspect with criminal cases, who had been put behind bars in local custody. However, the next day the man was found in an unconscious state and scant of breath. After admission and first-aid treatment, the victim was pronounced dead. Significantly, a half-piece of washrag was found blocked in his laryngeal cavity, inosculating with another piece concealed in his underpants. The psychological condition of criminal offenders is extremely complicated and varies with each individual. Effective administration and moderate counseling are not mutually exclusive with each other and both should afford the criminals a sense of calmness to face consequences.

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Scuba diver deaths due to air embolism: two case reports

Soud Lek. 2013 Spring;58(2):26-28

Barotraumas and decompression sickness are the two most well-known complications of diving. First presented case was 32 year-old male with recreational diver, who was found floating prone position on the bottom of sea in a depth of 33 m. He had been carried to the surface in a controlled ascent. Second case was a 39 year-old male experienced dive instructor in a diving school, after following an uneventful duration of dive was found unconscious with a floating supine position in a depth of 30 m and there were no signs of life when they were transported to the hospital. Extensive subcutaneous emphysema of the extremities was detected by palpation of the skin. In the autopsy diffuse gas bubbles like beads were seen in the coronary arteries and in ventricles, basilar artery and all of the cerebral arteries. The cause of death was attributed due to gas embolism and drowning. Keywords: barotraumas – diving – air embolism – autopsy.

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Estimation of blood alcohol concentration in deaths due to roadside accidents

Journal of Forensic and Legal Medicine  Volume 20, Issue 4, May 2013, Pages 300–304;  http://dx.doi.org/10.1016/j.jflm.2012.12.003

Like any other disease, accidents too are caused by interaction between agent, host, and environment. Human factors include age, (accidents most common between 10 and 24 years), sex, education, medical conditions (heart attack, impaired vision), fatigue, influence of alcohol and other drugs, lack of bodily protection (like helmets, seat belts) and psychosocial factors like lack of experience, impulsiveness, aggressiveness, defective judgment and delay in decisions. Drunken driving is an important risk factor in causing accidents. This study was focused on the status of alcohol consumption in relation with roadside accidents in northern India in the region of Amritsar. The present study was carried out in 100 cases alleged to have died of roadside accident and brought to the mortuary attached to the Department of Forensic Medicine and Toxicology, Government Medical College, Amritsar for autopsy examination. Blood samples were collected from the femoral vein and were tested for the presence of alcohol with steam distillation and titration method using potassium dichromate and sulfuric acid. In the present study, 23% of the fatal driver/pedestrian victims of roadside accidents were found to have consumed alcohol before accident. Most of the victims of road accident were from the age group 21–30 and 31–40 years. Most of the accidents occurred on straight roads instead of bends or intersections, more during daytime and weekends. 57% of the blood alcohol positives were between 100 mg% and 149 mg%. Majority of the victims of roadside accidents were motorcyclists and the striking vehicles were trucks and buses causing head & neck injuries in most of the victims. Death occurred within a few minutes in most of the cases.

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A fatal case of MAM-2201 poisoning

Forensic Toxicology May 2013

A 59-year-old man was found dead in his house, where three sachets containing herbal blends were found on a table. The sachet contents were analyzed by gas chromatography–mass spectrometry and found to contain [1-(5-fluoropentyl)-1H-indol-3-yl](4-methyl-1-naphthalenyl)methanone (MAM-2201). The deceased was subjected to forensic autopsy. There were neither external injuries nor endogenous diseases judged by macroscopic and microscopic observations. Liquid chromatography–electrospray ionization–tandem mass spectrometry was used to quantitate the concentrations of MAM-2201 in postmortem samples using deuterated MAM-2201 as internal standard. The MAM-2201 concentrations were: 12.4 ng/ml in whole blood; 18.1 ng/g in the liver; 11.2 ng/g in the kidney; 4.3 ng/g in the brain; and 1,535 ng/g in the adipose tissue. We concluded that the man’s death was caused by acute intoxication with MAM-2201. In addition, we propose that the adipose tissue is the specimen of choice to detect MAM-2201 in the unchanged form. To our knowledge, this is the first report of a fatal MAM-2201 poisoning case. In addition, this report is also the first to describe the distribution of the drug in postmortem human tissues and blood.

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EXPERIMENTS TO DETECT CLANDESTINE GRAVES FROM INTERPRETED HIGH RESOLUTION GEOPHYSICAL ANOMALIES

Meeting of the Americas in Cancún, Mexico 14 May 2013; Carlos Martin Molina et al [Poster]

This project refers to the search for clandestine sites where possibly missing people have been buried based on interpreted near surface high resolution geophysical anomalies. Nowadays, there are thousands of missing people around the world that could have been tortured and killed and buried in clandestine graves. This is a huge problem for their families and governments that are responsible to warranty the human rights for everybody. These people need to be found and the related crime cases need to be resolved. This work proposes to construct a series of graves where all the conditions of the grave, human remains and related objects are known. It is expected to detect contrasting physical properties of soil to identify the known human remains and objects. The proposed geophysical methods will include electrical tomography, magnetic and ground penetrating radar, among others. Two geographical sites will be selected to located and build standard graves with contrasting weather, soil, vegetation, geographic and geologic conditions. Forward and inverse modeling will be applied to locate and enhance the geophysical response of the known graves and to validate the methodology. As a result, an integrated geophysical program will be provided to support the search for clandestine graves helping to find missing people that have been illegally buried. Optionally, the methodology will be tested to search for real clandestine graves.

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Suicide due to four speargun shots: a case report

Am. J. Forensic Med. Pathol. In Press [May] 2013;  DOI: 10.1097/PAF.0b013e318288b0ed

In the literature, only a few cases of deaths related to the use of atypical firearms are present and even more rare are cases of suicide due to multiple lesions.In the present case, the authors report a rare occurrence of suicide due to 4 speargun shots, 3 to the chest and 1 to the head.A complete forensic approach led to attribute the death to acute cardiac failure due to hemopericardium after the injury of the left coronary artery.Scene investigation and autoptic findings allowed authors to hypothesize that injury to the head was a last attempt of suicide, elapsed during the progression of cardiac tamponade.With this report, the authors would like to share knowledge with the forensic community about speargun-related lesions distinguishing them from the very similar ones produced by cold steels.It also shows how it is possible to survive for some time after being shot by such a weapon.

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