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A sensitive and specific method for the determination of gamma-hydroxybutyric acid (GHB) in urine and serum is described. Prior to quantification by LC/MS in a Pauli-type ion trap, the molecule is converted by a fast and simple one-step procedure into its n-butyl ester derivative. Hexa-deutero GHB has been used as internal standard.
This study examines the effects of age, gender, a cold water medium versus warm water medium, and salinity on strontium levels determined in left ventricular blood in drownings. Significant differences in the amount of strontium absorbed into the bloodstream (p<0.001) were detected between individuals who drowned in fresh water versus those drowning in seawater, and between those drowning in cold water versus warm water (p=0.030). However, no significant differences were noted in the strontium concentrations of left ventricular blood according to gender or age.
We report a case of a juvenile male with muscle rigidity caused by cerebral palsy who experienced intraoperative sudden death due to pulmonary fat embolism after multiple muscle-release and tenotomy of the bilateral lower limbs. Data were obtained through review of the surgical and anesthesia records, as well as from autopsy and histopathological examination. All surgical procedures were performed within the same operation, beginning with the right lower limb and then proceeding with the left lower limb, with application of a pneumatic tourniquet to avoid intraoperative hemorrhage. Slight changes in the hemodynamics were noticed after release of the right tourniquet. Further, sudden onset of hypotension, severe bradycardia, and a marked decrease in percutaneously monitored oxygen saturation occurred just after release of the left tourniquet when the left limb was raised for casting. The patient died despite immediate and vigorous cardiopulmonary resuscitation. At autopsy performed 20 h after death, examination of the lungs revealed a pale surface, slight edema, and obvious fat droplets in the vessels at the cut surfaces. Histopathological examination with fat staining was notable for the presence of pulmonary fat embolism. These results suggest that restoration of venous return after removal of the tourniquet combined with massive fat embolism from dead spaces was the likely cause of death.
Disclosure of neurological disorders by neuropathological examination may be one important aim of forensic autopsies. There are insufficient data on human brain tissue preservation after prolonged postmortem periods. Here, we describe neuropathological findings in a brain of a 77-year-old woman that was fixed at autopsy 2 months after death. The body had been stored in a cooling chamber at 3 degrees C temperature. Gross inspection of the brain was satisfactorily possible. Histomorphology was excellently preserved. Many histochemical and immunohistochemical stains allowed satisfactory neuropathological evaluation of brain tissue and the diagnosis of Alzheimer's disease. Nevertheless, some immunohistochemical stainings repeatedly yielded negative or suboptimal results. We conclude that neuropathological examination of human brain tissue extracted from the skull and fixed after prolonged body storage in a cool environment is feasible for forensic diagnosis of neurological disorders even several months after death. However, in such cases the significance of negative immunohistochemical staining results must be interpreted with caution.
The distribution profile of infiltrated mast cell-subpopulations and eosinophils in the lung and heart sections of the patients who died of severe allergic hyperresponsiveness, was investigated. Four study groups were designed comprising 9 cases who died in systemic anaphylaxis (Group I), 10 asthmatic individuals whose death were assigned to acute and severe bronchial asthma (Group II), 10 asthmatic cases who died from non-immunological diseases (Group III). Twenty consecutive autopsies of non-allergic subjects who died of unnatural causes (Group IV) served as control group in this study. Utilizing antibodies against human tryptase and chymase and a double immunohistochemical staining method, we distinguished successfully all three subsets of mast cells (MC), MC-TC (containing both tryptase and chymase), MC-T (containing only tryptase) and MC-C (containing only chymase) types, subdivided on the basis of the protease compositions of their secretory granules. In order to immunostaining eosinophils, we used antibody to major basic protein as a marker. We also measured postmortem blood tryptase, specific and total serum IgE. The intriguing finding of this study was the marked differences of cellular composition in the lung between fatal anaphylaxis and asthma death. Significant augmentation of MCs infiltrated in lung and heart sections of anaphylaxis patients and drastic infiltration of bronchial eosinophils in asthmatic death and consequent release of their related inflammatory mediators might explain the differential expression of the associated symptoms in these two groups. The anaphylactic deaths did show neither emphysema nor significant mucous bronchial secretions whereas all asthmatic deaths did. The degree of pulmonary congestion and edema was also more severe in anaphylaxis. This corresponded with the histological findings and the location and number of mast cell-subsets and eosinophils in the different compartments of the lungs. We have demonstrated that the third type of mast cell MC-C is only found in the lungs in anaphylactic deaths. The practical consequence of our study will be that it is now possible to confirm a suspicion of anaphylaxis death not only by measurements of serum mast cell tryptase, but also by immunohistochemical methods.
After a mass fatality incident (MFI), all victims have to be rapidly and accurately identified for juridical reasons as well as for the relatives' sake. Since MFIs are often international in scope, Interpol has proposed standard disaster victim identification (DVI) procedures, which have been widely adopted by authorities and forensic experts. This study investigates how postmortem multislice computed tomography (MSCT) can contribute to the DVI process as proposed by Interpol. The Interpol postmortem (PM) form has been analyzed, and a number of items in sections D and E thereof have been postulated to be suitable for documentation by CT data. CT scans have then been performed on forensic cases. Interpretation of the reconstructed images showed that indeed much of the postmortem information required for identification can be gathered from CT data. Further advantages of the proposed approach concern the observer independent documentation, the possibility to reconstruct a variety of images a long time after the event, the possibility to distribute the work by transmitting CT data digitally, and the reduction of time and specialists needed at the disaster site. We conclude that MSCT may be used as a valuable screening tool in DVI in the future.
Detection of diatom frustules in bone marrow (diatom test) is used for diagnosing ante-mortem drowning where the usual signs of drowning are not present in dead bodies recovered from water. However, controversies over the reliability of diatom test results are continuing. There have been indications on the possibilities of diatoms entering into systemic circulation from atmospheric air, food and drink. While diatoms have been demonstrated in the gut content of edible marine forms such as shrimps and clams, the present study, for the first time, provides empirical evidence on the prevalence as well as abundance of diatom frustules in the samples of cooked non-vegetarian foodstuffs that impend human consumption in Kelantan, Malaysia. It is found that 50 g each of cleaned and cooked prawns and of clams impending human consumption contain about 8360 and 29,054 diatom frustules, respectively. A person accustomed to prawn and clam food would be ingesting an estimated 2 million diatoms in a single year. Considering the suggestion that detection of five diatom frustules in 10 g of bone marrow would suffice for concluding drowning as mode of death, and the fact that there is yet no proof that diatom frustules do not enter into the human systemic circulation through the digestive tract, the estimated number of diatom frustules routinely ingested acquires significance since entry of a few of such ingested frustules into the systemic circulation can lead to false positive test results. The findings of this research raise two important issues: first, population based routine food related diatom ingestion requires to be estimated, and, second, studies have to be initiated to categorically prove or disprove the possibility of entry of diatom frustules into the systemic circulation via the digestive tract.
OBJECTIVE: To compare the merit of four digestive methods (nitric acid plus hydrogen peroxide, proteinase K, nitric acid in Disorganization Can and Soluene-350) for extracting diatoms in order to choose the best digestive method for the diagnosis of drowning. METHODS: Liver, kidney and bone marrow of rabbits were minced and then digested by four digestive methods separately with the following indices compared: (1) time demanded for complete digestion; (2) degree of digestion for different tissues; (3) the reclaiming ratio of diatoms; (4) the degree of digestive destruction to diatoms. RESULTS: For sufficiently digesting the same tissue, the demanded times for the different methods ranked from the longest to the shortest were as follows: Soluene-350, proteinase K, nitric acid plus hydrogen peroxide, nitric acid in Disorganization Can. Nitric acid in Disorganization Can method and nitric acid plus hydrogen peroxide method digested the tissues more thoroughly than proteinase K, than Soluene-350 methods. For Cyclotella and Cybella, proteinase K method reclaimed most diatoms and nitric acid plus hydrogen peroxide method reclaimed less, while nitric acid in Disorganization Can and Soluene-350 methods reclaimed the least. For Navicula, the majority of diatoms could be extracted using proteinase K method, but only a few diatoms with other three methods. Under scanning electron microscopy (SEM), the structure of diatoms remained almost perfect after digestion with proteinase K, but destroyed to some extent with other three methods. CONCLUSION: This study demonstrated that different diatoms (in fresh or sea water) have different resistance to different digestive reagents. As far as the reliability and applicability of the diatom test is concerned, proteinase K method is of the best choice, nitric acid plus hydrogen peroxide can be its substitute. Soluene-350 cannot be used for extracting sea water diatoms.
We present a new tool for the estimation of the age of bloodstains, which could probably be used during forensic casework. For this, we used atomic force microscopy (AFM) for high-resolution imaging of erythrocytes in a blood sample and the detection of elasticity changes on a nanometer scale. For the analytic procedure we applied a fresh blood spot on a glass slide and started the AFM detection after drying of the blood drop. In a first step, an overview image was generated showing the presence of several red blood cells, which could easily be detected due to their typical "doughnut-like" appearance. The consecutively morphological investigations in a timeframe of 4 weeks could not show any alterations. Secondly, AFM was used to test the elasticity by recording force-distance curves. The measurements were performed immediately after drying, 1.5 h, 30 h and 31 days. The conditions were kept constant at room temperature (20 degrees C) and a humidity of 30%. The obtained elasticity parameters were plotted against a timeline and repeated several times. The elasticity pattern showed a decrease over time, which are most probably influenced by the alteration of the blood spot during the drying and coagulation process. The preliminary data demonstrates the capacity of this method to use it for development of calibration curves, which can be used for estimation of bloodstain ages during forensic investigations.
Free floating thrombus in the ascending aorta is an uncommon source of acute myocardial infarction. We report on two cases of young women who died of acute myocardial infarction caused by a free floating thrombus in the sinus of Valsalva obstructing the coronary arteries' ostia. The first case reports on a 30-year-old pregnant woman who anamnestically had episodes with short loss of consciousness and weakness. The second case presents a 37-year-old woman suffering from multiple sclerosis with no previous history of thrombotic events. The review of literature revealed a predominance of women (eight females and three males). Interestingly, the coronary arteries bear no preference concerning the right (RCA) or left coronary artery (LCA) being more often occluded by a free floating thrombus. Especially, younger women (mean age 45.5 years, range 30-59 years) with no history of cardiac symptoms and without atherosclerotic changes seem to be predispositioned. The hypothesis that thrombus formation in cases without plaque disruption may depend on an endothelial erosion which seems to be more common in younger women and promoted by a hyperthrombogenic state is supported by our two cases. A comprehensive literature search revealed, that these are the first two reports on a free floating thrombus being the cause of fatal acute myocardial infarction in a pregnant woman, respectively, a woman suffering from multiple sclerosis.
AIMS: To determine the frequency of extramedullary haematopoiesis (EMH) in sudden infant death and control cases, and to provide descriptive data on extramedullary haematopoietic islands and diffuse cells. METHODS: The study was carried out in 51 SIDS and 102 non-SIDS cases. Liver-tissue sections were stained with haematoxylin and eosin, and immunohistochemistry was done with streptavidin-biotin-peroxidase complexes by Polyclonal Haemoglobin "A" antibody. The prevalence of data was estimated statistically by Pearson chi-square test, descriptive statistics, and Kruskal-Wallis test. RESULTS: The Pearson chi-square values (CI: 95%; OR: 0.071-0.49; p: 0.0474) show higher frequency of EMH islands in SIDS than in controls. The number of diffuse extramedullary haematopoetic cells was higher in SIDS cases compared to non-SIDS (p=0.017). CONCLUSIONS: EMH is a frequent finding in SIDS cases, and this may be a consequence of anaemia associated with intrauterine hypoxia, or infections.
Aneurysms of the visceral arteries are a rather common feature appearing in 0.1-2% of the population. The clinical relevance of those anomalies varies a lot. The wide range of descriptions reaches from asymptomatic cases to fatalities in particular due to haemorrhages. The latter will be discussed in a case report concerning a 60-year-old man who collapsed at his work place and died 4 h after admission to the intensive care unit of a hospital nearby. The ruptured aneurysm remained undiagnosed in hospital and was found by autopsy. The case is presented and discussed in view of medico-legal questions.
The investigation of deaths due to the inhalation of volatile substances may be complicated by a lack of scene and autopsy findings. Mechanisms of death may not be determinable at autopsy, and there may be very few markers of inhalant abuse. A 21-year-old man is reported who died from the combined effects of methadone toxicity and toluene inhalation. Histological examination of the lungs revealed congestion and edema, as well as particles of blue, pigmented material within the interstitium and in macrophages. Scanning electron microscopy was undertaken, revealing that the particles contained granules that measured 0.15-0.2microm in diameter, within the range of mean particle sizes for inorganic paint pigments. Energy dispersive X-ray spectral analysis of the granules demonstrated a significant percentage of titanium (12%) confirming their origin from paint. Ancillary investigations such as electron microscopy and X-ray spectral analysis in cases of possible lethal volatile inhalation may prove useful adjuncts in determining the type of substance inhaled and in providing evidence of previous non-lethal episodes.
In clinical medicine, C-reactive protein (CRP) is extensively used as a general marker for immune system activation, and post-mortem applicability has been established [M.Q. Fujita, B.L. Zhu, K. Ishida, L. Quan, S. Oritani, H. Maeda, Serum C-reactive protein levels in postmortem blood-an analysis with special reference to the cause of death and survival time, Forensic Sci. Int. 130 (2002) 160-166; L. Uhlin-Hansen, C-reactive protein (CRP), a comparison of pre- and post-mortem blood levels, Forensic Sci. Int. 124 (2001) 32-35]. We have analysed the routine use of CRP in non-selected cases. Scarcity of blood available for analysis is a common problem in forensic investigation, and in response to this we have developed a method using liver as a source. In 50 consecutive autopsy cases, we have evaluated method, validated results and discussed their interpretation. In three cases the analysis was not possible. For each of the remaining cases (n=47) we have analysed whole blood, serum and/or liver samples. 57% (n=25) had serum CRP > 10 mg/L. Serum levels were higher than in whole blood or liver. CRP levels in serum and whole blood samples were stable in more than one month after death, making storage for later analysis possible. Liver levels peaked at one week, but after one month putrefaction was obvious. CRP levels were independent of the post-mortem interval. The use of liver as a source has not yet been described in literature. Our results in liver samples correlate well with plasma results, and liver is a good post-mortem alternative when blood is not available. We conclude that CRP measurements are easy, viable and inexpensive in a forensic setting, and that the number of cases with CRP elevation is high in a non-selected forensic material. In cases of doubt, marked elevation of CRP is an indicator of natural mode of death, and in cases of trauma, it indicates vital reaction. It can be used as a pre-autopsy screening, leading to a more extensive search for diseases not easily diagnosed, such as sepsis or ketoacidosis.
This case study documents the nearly complete consumption of adult human remains by two domestic dogs (Canis familiaris) inside a residence. While scavenging behavior has been observed for coyotes, wolves, hyenas and other canines in natural outdoor environments, little information is available concerning canine scavenging of human remains in an indoor setting. In this case, the dogs were confined with the body inside a residence for approximately 1 month. The impact of intrinsic and extrinsic factors on canine scavenging behavior and the postmortem interval are examined, such as clothing on the body, perimortem trauma, drug ingestion and the availability of alternative food sources. While cases of canine scavenging of their owners are scant in the literature, this phenomenon is probably not uncommon, particularly among the elderly and indigent who live alone with pets and are socially isolated.
Acute interstitial pneumonitis (AIP), also known as Hamman-Rich syndrome, is a distinct type of idiopathic interstitial pneumonia affecting patients of both genders without pre-existing lung diseases. We describe the case of a fulminant form of AIP and discuss the pathophysiological mechanisms of AIP with reference to the histological pattern. A 15-year-previously-healthy male boy presented to the Hospital with a 6-day history of malaise, fever and cough. The clinical prodromes were followed by the acute onset of increasing shortness of breath rapidly progressing in acute respiratory failure. Chest X-ray demonstrated bilateral diffuse airspace opacification; the high resolution CT confirmed the presence of bilateral, symmetric diffuse ground-glass attenuation. The patient was admitted to the intensive care unit, but died after few hours. An autopsy was performed within 24h. The histological examination of lung specimens showed a pattern of diffuse alveolar damage. immunohistochemical, microbiological and toxicological tests were also carried out. The clinical presentation, the histological findings and the exclusion of infective, traumatic, toxic and metabolic causes of acute respiratory distress syndrome (ARDS) allowed us to conclude that the boy was affected by AIP. In conclusion, AIP is a diagnosis of exclusion. It has a mortality rate ranging about 50%, despite mechanical ventilation. In fatal cases of AIP diagnosis can be based on clinical presentation, radiological, histological and microbiological findings and can be further confirmed by immunohistochemical analysis.
The differentiation of SIDS from accidental or inflicted suffocation may be impossible without corroborating findings from the death scene or autopsy or in the absence of a confession from a perpetrator. Pulmonary intra-alveolar hemorrhage (PH) has been proposed as a potential clue to suffocation, but none of the previous studies on this topic have limited SIDS cases to those who were in a safe sleep environment, in which all were found supine and alone on a firm surface with their heads uncovered. Our aims are to: (1) compare PH in SIDS cases found in a safe sleep environment to a control group comprised of infants whose deaths were attributed to accidental or inflicted suffocation and (2) assess the effect of age, CPR, and postmortem interval (PMI), with regard to the severity of PH in this subset of safe-sleeping SIDS cases. We conducted a retrospective study of all postneonatal cases accessioned by the Office of the Medical Examiner in San Diego County, California who died of SIDS or suffocation between 1999 and 2004. A total of 74 cases of sudden infant death caused by SIDS (34 cases as defined above, comprising 8% of the total SIDS cases), accidental suffocation (37), and inflicted suffocation (3) from the San Diego SIDS/SUDC Research Project database were compared using a semiquantitative measure of pulmonary intra-alveolar hemorrhage. The most severe (grade 3 or 4) PH occurred in 35% of deaths attributed to suffocation, but in only 9% of the SIDS cases. Age, duration of CPR attempts and PMI had no effect on the severity of PH in SIDS. Our results indicate that the severity of PH cannot be used independently to differentiate SIDS from suffocation deaths. Each case must be evaluated on its own merits after thorough review of the medical history, circumstances of death, and postmortem findings.
Two cases are reported where significant narrowing of the upper airway in association with narcotic administration resulted in respiratory compromise and death. Case 1: A 29-year-old woman with upper airway narrowing due to tonsillar enlargement from an Epstein-Barr infection was admitted to hospital, administered morphine and left in a room on her own with the door closed. She was found dead several hours later. At autopsy there was significant narrowing of the upper airway due to tonsillomegaly with a blood morphine level of 0.16 mg/L. Case 2: A 48-year-old woman with severe narrowing of her glottic inlet from recurrent squamous cell carcinoma and an intravenous drug taking history was found dead at her home. At autopsy there was evidence of recent and remote intravenous drug administration with marked narrowing of the glottis due to a recurrent tumor with a blood morphine level of 0.48 mg/L. In both cases, death was due to the effects of severe upper airway narrowing in combination with the respiratory depressant actions of morphine. Additional exacerbating factors may have included muscle weakness, drowsiness and reduced clearance of airway secretions from the effects of morphine. Narcotic administration in individuals with compromised upper airways should be undertaken extremely circumspectly and hospital protocols should ensure constant surveillance if this has been undertaken. Individuals who self administer narcotics should also be made aware of the dangers if there is coincidental upper airway narrowing. Toxicological evaluation in fatal cases of upper airway narrowing/stenosis may be extremely useful in revealing compounding factors such as opiate administration.