A concept study on identification and attribution profiling of chemical threat agents using liquid chromatography-mass spectrometry applied to Amanita toxins in food

Accidental or deliberate poisoning of food is of great national and international concern. Detecting and identifying potentially toxic agents in food is challenging due to their large chemical diversity and the complexity range of food matrices. A methodology is presented whereby toxic agents are identified and further characterized using a two-step approach. First, generic screening is performed by LC/MS/MS to detect toxins based on a list of selected potential chemical threat agents (CTAs). After identifying the CTAs, a second LC/MS analysis is performed applying accurate mass determination and the generation of an attribution profile. To demonstrate the potential of the methodology, toxins from the mushrooms Amanita phalloides and Amanita virosa were analyzed. These mushrooms are known to produce cyclic peptide toxins, which can be grouped into amatoxins, phallotoxins and virotoxins, where α-amanitin and β-amanitin are regarded as the most potent. To represent a typical complex food sample, mushroom stews containing either A. phalloides or A. virosa were prepared. By combining the screening method with accurate mass analysis, the attribution profile for the identified toxins and related components in each stew was established and used to identify the mushroom species in question. In addition, the analytical data was consistent with the fact that the A. virosa specimens used in this study were of European origin. This adds an important piece of information that enables geographic attribution and strengthens the attribution profile.     

Postnatal ontogenesis of the tibia. Implications for age and sex estimation

The growth of five variables of the tibia (diaphyseal length, diaphyseal length plus distal epiphysis, condylo-malleolar length, sagittal diameter of the proximal epiphysis, maximum breadth of the distal epiphysis) were analysed using polynomial regression in order to evaluate their significance and capacity for age and sex determination during and after growth. Data were collected from 181 (90♂ and 91♀) individuals ranging from birth to 25 years of age and belonging to three documented collections from Western Europe. Results indicate that all five variables exhibit linear behaviour during growth, which can be expressed by a first-degree polynomial function. Sexual significant differences were observed from age 15 onward in the two epiphysis measurements and condylo-malleolar length, suggesting that these three variables could be useful for sex determination in individuals older than 15 years. Strong correlation coefficients were identified between the five tibial variables and age. These results indicate that any of the studied tibial measurements is likely to serve as a useful source for estimating sub-adult age in both archaeological and forensic samples.     

Traumatic bilateral vertebral artery dissection

Traumatic vertebral artery dissection is not often seen by forensic pathologists, and cases investigated are scarce in the forensic literature. We present the case of a 40-year-old woman cyclist who was struck by a car while wearing a helmet, and was neurologically near normal immediately thereafter at Emergency. She presented 48 h later with acute right hemiparesis, decreasing level of consciousness, and unsteadiness. CT revealed massive cerebellar infarction. CT angiography was normal. The patient died in coma 7 days after injury and autopsy revealed bilateral edematous cerebellar infarction and bilateral vertebral artery dissection. Rotational neck injury and mural tear in the wall of the Atlantic parts of both vertebral arteries is suggested as the possible mechanism of the arterial injury. Head and neck injuries are reported as a precipitating cause of vertebral artery injury. The possible influence of trauma may be further underestimated if longer intervals between vessel dissection and ischemia occur. The current case illustrates that "talk-and-die" syndrome may be due to occult vertebral artery dissection, possibly bilateral. In forensic cases of delayed death after mild trauma to the head and neck, the vertebral arteries should be examined for the cause of death.     

Raman spectroscopy and microspectrophotometry of reactive dyes on cotton fibres: Analysis and detection limits

A collaborative study on Raman spectroscopy and microspectrophotometry (MSP) was carried out by members of the ENFSI (European Network of Forensic Science Institutes) European Fibres Group (EFG) on different dyed cotton fabrics. The detection limits of the two methods were tested on two cotton sets with a dye concentration ranging from 0.5 to 0.005% (w/w). This survey shows that it is possible to detect the presence of dye in fibres with concentrations below that detectable by the traditional methods of light microscopy and microspectrophotometry (MSP). The MSP detection limit for the dyes used in this study was found to be a concentration of 0.5% (w/w). At this concentration, the fibres appear colourless with light microscopy. Raman spectroscopy clearly shows a higher potential to detect concentrations of dyes as low as 0.05% for the yellow dye RY145 and 0.005% for the blue dye RB221. This detection limit was found to depend both on the chemical composition of the dye itself and on the analytical conditions, particularly the laser wavelength. Furthermore, analysis of binary mixtures of dyes showed that while the minor dye was detected at 1.5% (w/w) (30% of the total dye concentration) using microspectrophotometry, it was detected at a level as low as 0.05% (w/w) (10% of the total dye concentration) using Raman spectroscopy. This work also highlights the importance of a flexible Raman instrument equipped with several lasers at different wavelengths for the analysis of dyed fibres. The operator and the set up of the analytical conditions are also of prime importance in order to obtain high quality spectra. Changing the laser wavelength is important to detect different dyes in a mixture.     

Clinical and forensic examinations of glycemic marker 1,5-anhydroglucitol by means of high performance liquid chromatography tandem mass spectrometry

Postmortem diagnosis of diabetes and a diabetic coma can be difficult because of the lack of characteristic morphological findings. 1,5-Anhydroglucitol (1,5-AG), the 1-deoxy form of glucose, competes with glucose for reabsorption in the kidneys. Therefore, diabetics with a permanent hyperglycemia show significantly lower serum concentrations of 1,5-AG than non-diabetics. A liquid chromatography-mass spectrometric method for the determination of 1,5-AG in serum and postmortem blood was developed and validated according to international guidelines. Linearity was given between 1μg/ml and 50μg/ml. Recovery rates ranged between 70.8% and 89.8%, the limit of quantification of the procedure was 0.20μg/ml, limit of quantification was 0.55μg/ml. Serum of 199 diabetics and 116 non-diabetics and femoral blood of 31 diabetic and 27 non-diabetic deceased was measured. Average concentrations were significantly (p<0.001) higher in non-diabetics compared to diabetics ante and postmortem. Seven of the diabetics may have died because of a hyperglycemic coma indicated by a sum formula of Traub>450mg/dl. 1,5-AG average concentrations in these deceased were not significantly different to diabetics which did not die because of a diabetic coma. Concentrations of 1,5-AG give a hint for not well controlled diabetes antemortem and postmortem and can be assumed as an additional and alternative information postmortem to the measurement of HbA1c or fructosamine.     

Fatal tolperisone poisoning: autopsy and toxicology findings in three suicide cases

Tolperisone (Mydocalm) is a centrally acting muscle relaxant with few sedative side effects that is used for the treatment of chronic pain conditions. We describe three cases of suicidal tolperisone poisoning in three healthy young subjects in the years 2006, 2008 and 2009. In all cases, macroscopic and microscopic autopsy findings did not reveal the cause of death. Systematic toxicological analysis (STA) including immunological tests, screening for volatile substances and blood, urine and gastric content screening by GC-MS and HPLC-DAD demonstrated the presence of tolperisone in all cases. In addition to tolperisone, only the analgesics paracetamol (acetaminophen), ibuprofen and naproxen could be detected. The blood ethanol concentrations were all lower than 0.10 g/kg. Tolperisone was extracted by liquid-liquid extraction using n-chlorobutane as the extraction solvent. The quantification was performed by GC-NPD analysis of blood, urine and gastric content. Tolperisone concentrations of 7.0 mg/l, 14 mg/l and 19 mg/l were found in the blood of the deceased. In the absence of other autopsy findings, the deaths in these three cases were finally explained as a result of lethal tolperisone ingestion. To the best of our knowledge, these three cases are the first reported cases of suicidal tolperisone poisonings.     

Housing instability is as strong a predictor of poor health outcomes as level of danger in an abusive relationship: findings from the SHARE Study

Advocates, clinicians, policy makers, and survivors frequently cite intimate partner violence (IPV) as an immediate cause of or precursor to housing problems. Research has indicated an association between homelessness and IPV, yet few studies examine IPV and housing instability. Housing instability differs from homelessness, in that someone experiencing housing instability may currently have a place to live but faces difficulties with maintaining the residence. We present baseline findings from a longitudinal cohort study of 278 female IPV survivors with housing as a primary concern. Our analysis indicates the greater the number of housing instability risk factors (e.g., eviction notice, problems with landlord, moving multiple times), the more likely the abused woman reported symptoms consistent with PTSD (p < .001), depression (p < .001), reduced quality of life (p < .001), increased work/school absence (OR = 1.28, p < .004), and increased hospital/emergency department use (OR = 1.22, p < .001). These outcomes persist even when controlling for the level of danger in the abusive relationship and for survivors' drug and alcohol use. Importantly, both housing instability and danger level had stronger associations with negative health outcomes than other factors such as age, alcohol, and drug use; both make unique contributions to negative health outcomes and could contribute in different ways. Housing instability is an important and understudied social determinant of health for IPV survivors. These findings begin to address the literature gap on the relationship between housing instability, IPV, and survivors' health, employment, and utilization of medical care services.     

Intimate partner violence against older women in Germany: prevalence and associated factors

Violence against women is a recognized human rights and public health issue, with significant impacts on women's life and health. Until now, several studies, most of them relying on small scale samples, have explored the prevalence and health effects of intimate partner violence against older women, whereas few have examined what actually puts older women at risk of intimate partner violence. This study is based on a secondary analysis of the first national survey on violence against women in Germany, looking at the prevalence and associated factors for physical and for sexual violence by the current partners of women aged 50 to 65 and women aged 66 to 86 years. The prevalence of violence in women's current relationships was 12% and 5%, respectively. In both age groups, women who had experienced violence during childhood and nonpartner physical or sexual violence after the age of 16 had higher odds of experiencing current partner violence. Current partner violence was associated only with women and their partner's level of education and women's vocational training among women aged 66 to 86 years. Relationships where one or both partners drank heavily in recent months were associated with higher odds of violence among women aged 50 to 65. Future studies on intimate partner violence need to recognize that women above reproductive risk are also at risk of current partner violence.     

Variations in wounding by relationship intimacy in homicide cases

There are numerous examples in the homicide literature of a presumed connection between the victim-offender relationship and the manner, extent, and body location of wounds inflicted in homicides. The current study examined variations in wounding patterns according to the intimacy of the victim-offender relationship in a sample of urban homicides to explore the investigative utility of this information in an average homicide event. The findings demonstrated that victims who had a current or former intimate relationship with their offender were more likely to receive wounds to the face and be injured with a weapon from the scene compared with all other relationship groups, whereas injury to the head and use of manual violence were more likely among intimates and family/friends compared with acquaintances or strangers. However, the groups did not significantly differ in terms of the overall amount of wounds inflicted. Implications of the findings and suggested areas of future research are discussed.     

Mediating effects of stalking victimization on gender differences in mental health

Studies suggest that stalking victimization may have a serious mental health impact. The present article investigates gender differences in mental health and possible mediating effects of stalking victimization in a community sample. The study includes a postal survey of 665 German community residents on the experience of stalking and various mental health indicators. In a majority of mental health scales, women score poorer than men, and a higher percentage of women fulfill criteria for a current mental disorder and used psychotropic medication. However, effects of gender decrease to a nonsignificant level when stalking victimization is entered into the respective models. Furthermore, associations of stalking victimization with poor mental health, psychosocial functioning, and use of medication are largely comparable across gender.