Immunohistochemical contribution to the study of morphine metabolism in Calliphoridae larvae and implications in forensic entomotoxicology

Morphine was detected by immunohistochemistry on sections of third stage larvae of Calliphora vomitoria (Diptera, Calliphoridae) reared on minced beef meat previously treated with morphine hydrochloride. The detection was performed with an avidin-biotin-peroxidase-complex method. Positive specimens showed specific staining of the haemolymph and a more intense immunoreaction in an area located at the limit between exocuticle and endocuticle. These results constitute an evidence of morphine accumulation inside the cuticle of Diptera larvae during their development. During the pupariation, the larval cuticle is transformed into the sclerotized puparium. This study consequently points out the possibilities of analyzing empty pupariae when suitable tissues or living necrophagous insects are absent.     

The contract murderer: patterns, characteristics, and dynamics

A case of an independent professional contract murderer, who killed over 100 people, is reported. After eluding law enforcement for 30 years, the subject killed several associates who he believed could implicate him in various crimes. These homicides eventually led to his arrest, since the victims were individuals who could be linked to him. This hit man had a background of poverty and childhood abuse but, as an adult, had pursued a middle-class lifestyle and kept his family totally separate from his criminal career. In addition, he had a number of characteristics that helped him carry out his crimes in a highly planned, methodical, and organized manner: he had adept social judgment; personality traits of orderliness, control, and paranoid vigilance; useful defense mechanisms of rationalization and reframing; and an exceptional ability to encapsulate emotions. This case is discussed within the context of contract murder, a crime that occurs relatively frequently and is probably increasing; yet it often goes undetected, the arrest rate is low, and the offender is rarely studied.     

Deposition of 7-aminoflunitrazepam and flunitrazepam in hair after a single dose of Rohypnol

In recent years, there has been a notable increase in the number of reports on drug-facilitated sexual assault. Benzodiazepines are the most common so-called "date-rape" drugs, with flunitrazepam (Rohypnol) being one of the most frequently mentioned. The aim of this study was to determine whether flunitrazepam and its major metabolite 7-aminoflunitrazepam could be detected in hair collected from ten healthy volunteers after receiving a single 2 mg dose of Rohypnol using solid phase extraction and NCI-GC-MS. Such data would be of great importance to law enforcement agencies trying to determine the best time interval for hair collection from a victim of drug-facilitated sexual assault in order to reveal drug use. Ten healthy volunteers (eight women and two men, 21 to 49 years old) participated in the study. The following hair samples were collected from each volunteer: one before flunitrazepam administration, and 1, 3, 5, 14, 21, and 28 days after. In five volunteers, 7-aminoflunitrazepam was detected 24 h after flunitrazepam administration and remained in hair throughout the entire 28-day study period (0.6-8.0 pg/mg). In two cases, 7-aminoflunitrazepam appeared in hair 21 days after drug intake (0.5-2.7 pg/mg), and in two subjects 14 days later (0.5-5.4 pg/mg). In one volunteer, 7-aminoflunitrazepam was detected on day 14 and 21 but concentrations were below the quantitation limit. Flunitrazepam was detected in some samples but all concentrations were below the quantitation limit (0.5-2.3 pg/mg).     

Validation of short tandem repeats (STRs) for forensic usage: performance testing of fluorescent multiplex STR systems and analysis of authentic and simulated forensic samples

The amplification and typing conditions for the 13 core CODIS loci and their forensic applicability were evaluated. These loci are CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, and D21S11. Results were obtained using the multiplex STR systems AmpFlSTR Profiler Plus and AmpFlSTR COfiler (Applied Biosystems, Foster City, CA), GenePrint PowerPlex (Promega Corporation, Madison, WI), and subsets of these kits. For detection of fluorescently labeled amplified products, the ABI Prism 310 Genetic Analyzer, the ABI Prism 377 DNA Sequencer, the FMBIO II Fluorescent Imaging Device, and the Fluorlmager were utilized. The following studies were conducted: (a) evaluation of PCR parameter ranges required for adequate performance in multiplex amplification of STR loci, (b) determination of the sensitivity of detection of the systems, (c) characterization of non-allelic PCR products, (d) evaluation of heterozygous peak intensities, (e) determination of the relative level of stutter per locus, (f) determination of stochastic PCR thresholds, (g) analysis of previously typed case samples, environmentally insulted samples, and body fluid samples deposited on various substrates, and (h) detection of components of mixed DNA samples. The data demonstrate that the commercially available multiplex kits can be used to amplify and type STR loci successfully from DNA derived from human biological specimens. There was no evidence of false positive or false negative results and no substantial evidence of preferential amplification within a locus. Although at times general balance among loci labeled with the same fluorophore was not observed, the results obtained were still valid and robust. Suggested criteria are provided for determining whether a sample is derived from a single source or from more than one contributor. These criteria entail the following: (a) the number of peaks at a locus, (b) the relative height of stutter products, and (c) peak height ratios. Stochastic threshold levels and the efficiency of non-templated nucleotide addition should be considered when evaluating the presence of mixtures or low quantity DNA samples. Guidelines, not standards, for interpretation should be developed to interpret STR profiles in cases, because there will be instances in which the standards may not apply. These instances include (a) a primer binding site variant for one allele at a given locus, (b) unusually high stutter product, (c) gene duplication, and (d) translocation.     

Entrapment in small, enclosed spaces: a case report and points to consider regarding the mechanism of death

The mechanism of death due to confinement in an enclosed space is usually ascribed to asphyxia from oxygen deprivation. We report the case of the decomposed remains of a 23-year-old man discovered in an unused industrial size refrigerator in which the mechanism of death is heatstroke. The investigation of the death indicates the subject most likely voluntarily entered the refrigerator and for unknown reasons, closed the door. Injuries identified at autopsy and damage to the inside of the structure indicate he struggled to exit the cabinet. The autopsy shows no significant natural disease processes and toxicology studies were negative. The diagnosis of heat stroke typically rests on the evaluation of multiple features, including the age and size of the decedent, the ambient temperature, the medical history of the decedent, whole body hydration, body fat content, alcohol and drug use, medication history, general physical condition, and many other factors. The diagnosis of heatstroke due to confinement in an enclosed container requires evaluation of the heat stress of the container, the heat strain experienced by the individual, autopsy findings suggesting signs of a struggle to exit the container, and other factors. In all such cases, a careful death investigation with correlation of autopsy findings is required to accurately determine the mechanism and cause of death. We suggest that for all such deaths, physiological and environmental factors promoting hyperthermia and heatstroke be considered as a possible mechanism of death, along with those associated with the more obvious danger of asphyxiation.     

An investigation of the psychological characteristics of stalkers: empathy, problem-solving, attachment and borderline personality features

This study examined the psychological characteristics of a sample of self-reported stalkers in comparison with a control group, on measures of empathy, problem-solving skills, attachment, and borderline personality features. Stalkers were identified by their endorsement of specific behavioral items, consistent with a widely adopted definition of stalking, denoting behaviors that: (a) are repeatedly directed toward an identified target; (b) are intrusive and unwanted; and (c) evoke fear in the victim. Stalkers scored significantly higher than controls on measures of insecure attachment and borderline personality features, suggesting that the stalking group demonstrates a general pattern of inadequate interpersonal attachment, has limited abilities to form and maintain appropriate relationships, is emotionally labile and unstable, and experiences ambivalence regarding their interpersonal relationships. Treatment implications are discussed herein.     

Examination of variation in sternal rib end morphology relevant to age assessment

The morphology of the sternal end of the right fourth rib has been proffered as an accurate age assessor in skeletonized individuals of both sexes. This technique was tested for its applicability on left and right II, III, V-IX. Tests were performed between phase scores obtained from right and left ribs; right rib IV phase scores and scores obtained from the others in the right rib series; and between right rib IV scores and a composite score composed of the average of an individual's phase scores (omitting rib IV). Left ribs IV-IX were found not to vary significantly from their right counterparts. Although only right rib II was found to vary significantly from rib IV, use of the other ribs in the series should be undertaken with caution due to questions concerning their statistical significance. A composite score is therefore recommended for use instead.