Spectrophotometric evaluation of the age of bruises in children: measuring changes in bruise color as an indicator of child physical abuse.

Physical abuse of children is a severe social problem and is usually identified by the presence of bruises of various ages. The visual appearance of bruises is widely used to identify victims of abuse. Therefore, to objectively evaluate the age of bruises, we used a spectrophotometer to examine 86 bruises that occurred accidentally in healthy child volunteers, with consent from appropriate guardians. The bruise color was measured using a spectrophotometer and plotted using the CIE-L*a*b* color system, a method that expresses color numerically. The differences [Delta] in L* (lightness), a* (red and green content) and b* (yellow and blue content) color values relative to neighboring healthy/unbruised skin were measured for 7-10 days until the bruise disappeared. A characteristic pattern was observed in 21 bruises; DeltaL* increased from the negative peak and returned to baseline, Deltaa* decreased from the positive peak to baseline, and Deltab* increased above baseline and then slowly returned to baseline. The pattern of these color changes could be classified into three phases according to the time between bruising and the peak values for DeltaL* (negative peak, 38.9 +/- 19.5 hours), Deltaa* (positive peak, 37.6 +/- 21.7 hours), and Deltab* (negative and positive peak, 43.5 +/- 18.7 and 132.6 +/- 40.4 hours). Thus, spectrophotometric measurement of the color of bruises is helpful to estimate the approximate age of bruises and to distinguish between old and new bruises. This objective method could be introduced to clinical practice and social care to evaluate possible cases of child abuse.

Vertebral arterial injury due to fatal dog bites.

Dog bite-related injuries and fatalities are major public health problem worldwide. The authors present a case of a Japanese Tosa-dog bite-related fatality in an old woman who died from a vertebral arterial laceration with the C5 vertebral fracture. This was an uncommon type of injury that dog bites injured the vertebral artery only without damage to carotid artery. The identity of the Tosa-dog as an offending dog in this case was unsuccessful by a DNA analysis. Then the detection of the offending dog was made by comparisons of the dental casts of the dog with the victim's wounds. This case report describes making dental cast of the dog, and statistical data on dog-bite incidents in Japan.

A rare case of sudden death due to thrombus from the internal iliac vein.

A 40-year-old healthy housewife died after joining a special summer camp for her diabetic daughter. The victim sat for 4h 40 min while traveling by car on the day before her death. Autopsy revealed she died from a pulmonary thromboembolism (PTE), whereby portions of a blood-clot that had been formed in the internal iliac veins detached and traveled to the lungs, blocking the pulmonary blood flow. The leg veins are the most common source of thromboembolism; PTE caused by thromboemboli from the internal iliac veins is rare. We discuss the mechanisms of the present victim's death, and the conditions and risk factors involved in PTE.

Immunohistochemical demonstration of the distribution of chloroquine (CQ) and its metabolites in CQ-poisoned mice.

Chloroquine (CQ) distribution in tissues of acutely poisoned mice was demonstrated by immunohistochemistry using anti-CQ polyclonal antibodies (PAC). PAC recognized 4-amino-7-chloro-quinoline structure and sufficiently reacted with CQ and CQ's metabolite bisdesethyl-chloroquine. In the brain, CQ and its metabolites (CQs) localized in the region of the choroids plexus, indicating an important role in the blood-cerebrospinal barrier system. In the heart, most regions showed diffused positive staining, and relatively strong reaction was observed in Purkinje cells, indicating an important role in acute CQ toxicity. In the lungs, CQs were observed in the bronchial epithelium, type II pneumocytes, and on the surface of alveolar walls. It was suggested that CQs were excreted to the alveolar wall with surfactant phospholipids, which are produced by type II pneumocytes. In the liver, CQs were concentrated in the centrolobular area rather than in the periportal area, in agreement with CQ's metabolic pathway. In the kidneys, tubular cells were strongly stained compared to glomerular capsules, and the distal part of renal tubules was better stained than the proximal tubules. These findings suggested that CQs were predominantly excreted or reabsorbed through the distal tubules and the collecting duct. Distribution of CQs in tissues presented here were mostly consistent with the physico-chemical properties of CQ and its metabolites. However, the elucidation of CQs' localization in Purkinje cells remains open. Further experimental studies at the level of microorganella will be needed to clarify the present result.

[Two cases of diphenhydramine related suicide].

Two suicidal cases associated with ingestion of diphenhydramine (DPH) were reported. Case 1 is a typical DPH overdose case of a young man with the blood DPH concentration of 12.2 microg/ mL. Case 2 is a double suicide of a man and a woman. They ingested DPH and fell asleep in a vehicle which had a cooking clay charcoal stove. Their blood DPH concentrations were 0.4 and 0.7 microg/mL, which were high enough to make them sleep. Their cause of death, however, was carbon monoxide poisoning with blood CO-Hb concentration of 14 and 19%. DPH is a low toxic agent and is available as an OTC drug in Japan. Similar fatal cases can be expected to happen in Japan.

Sudden death of a young woman due to aortic dissection caused by Turner's syndrome.

A 24-year-old woman was found dead in her bed. There had been an episode of fainting with cervicodynia 1 day before death but no significant past medical history, except for menstrual irregularities. Post-mortem examination revealed that death was due to hemopericardium caused by rupture of the ascending aorta by thoracic aortic dissection (Stanford type A). Microscopically, weakness of the aorta was due to cystic medial necrosis. On external examination, short stature, a short neck and multiple pigmented nevi were observed, while internal examination revealed coarctation of the aorta and funicular ovaries. Examination of the X chromatin showed a decrease in numbers of Barr bodies in the tissues, and a 45,X/46,XX mosaicism was suspected. It is concluded that the cause of death was aortic dissection due to Turner's syndrome.

An accidental death due to Freon 22 (monochlorodifluoromethane) inhalation in a fishing vessel.

A case of accidental Freon 22 (monochlorodifluoromethane) poisoning in a fishing vessel is reported. Forensic autopsy revealed severe pulmonary edema and congestion (left lung; 576 g, right lung; 740 g). GC-MS analysis clearly showed that the deceased inhaled Freon 22 gas prior to his death. Freon 22 concentration was 169+/-7.0 microg/ml in the heart blood. The distribution pattern of Freon 22 in tissue samples was similar to that in previously reported cases. The brain had the highest concentration of Freon 22 followed by the spleen, liver, kidney and lung, respectively. Histopathologically, Oil red O staining of the liver showed many small, positive red areas in the cytosol, which have been reported in other cases of Freon 22 poisoning. However, Schmorl staining revealed that most areas of Oil red O positivity were lipofuscin granules. Lipofuscin in the liver, which closely relates to aging and other cell stresses, could have a relevance to Freon 22 exposure, but further experimental studies are needed to confirm it.

Postmortem cytokine levels and severity of traumatic injuries.

The relationship between postmortem serum cytokine levels and severity of traumatic injuries was studied. The postmortem serum levels of interleukin-6 (IL-6) and interleukin-8 (IL-8) of 131 victims who died from traumatic injury were measured by an enzyme-linked immunosorbent assay method and compared with scores of total abbreviated injury scale (total AIS) and injury severity score (ISS) calculated from detailed autopsy reports. A significant positive correlation was observed between IL-6 and total AIS (rs=0.4508, p<0.0001), between IL-6 and ISS (rs=0.3337, p<0.0001), between IL-8 and total AIS (rs=0.6593, p<0.0001), and between IL-8 and ISS (rs=0.5305, p<0.0001). The significant correlation between cytokine levels and anatomical traumatic severity indicated that the cytokine levels are useful objective indexes of traumatic severity. In addition, the total AIS is a suitable marker to evaluate traumatic severity as the coefficient of correlation between the cytokine levels and the total AIS was higher than that for the ISS values.

Neuropathology of human immunodeficiency virus infection: a forensic autopsy study in Dar Es Salaam, Tanzania.

The objective of this study was to examine the neuropathological changes in the brain of patients infected with human immunodeficiency virus (HIV) in the Tanzanian capital Dar Es Salaam, and investigate whether the prevalence of different forms of HIV-related neuropathology varies from other countries. The subjects were patients with risk factors for HIV infection in whom forensic autopsies were performed between 1997 and 1999. In Dar Es Salaam, forensic autopsy constitutes more than 90% of all autopsies, because hospital autopsy is limited due to socio-cultural and religious reasons. HIV infection was identified in 52 of 143 patients selected from forensic autopsies. Neuropathological findings were observed in 31 of 52 HIV-infected patients; these include lymphocytic meningitis 19, bacterial meningitis 3, tuberculous brain abscess 3, cryptococcal meningitis 3, basal ganglia calcification 3, and toxoplasma encephalitis 1. HIV encephalitis, lymphoma, and cytomegalovirus encephalitis could not be found in this study. Whereas the findings should be interpreted cautiously because of possible autopsy bias and a low percentage of cases examined compared to the total number of HIV-infected patients in Tanzania, our observations provide information on the likely diagnostic possibilities to be considered in the evaluation and management of HIV-infected patients with neurological symptoms in Tanzania. In the face of decreased hospital autopsy, most studies have focused mainly on the end-stage HIV disease; forensic autopsy is a potential source of materials for studies on HIV disease spectrum at different stages.

Estimation of early postmortem intervals by a multiple regression analysis using rectal temperature and non-temperature based postmortem changes.

Five general methods based on rectal temperature and a multiple regression analysis using rectal temperature and non-temperature based postmortem changes were applied to 212 postmortem cases of within 24h postmortem (PM) intervals. Non-temperature based postmortem changes of rigidity, hypostasis and corneal turbidity were numerically categorized and used with rectal temperatures as four statistical variables in the multiple regression analysis. The correlation coefficient values between true and calculated postmortem intervals were 0.78-0.82 in the five general methods based on rectal temperature. The multiple regression analysis produced a multiple correlation coefficient value of 0.89 and according to the error ranges of the PM intervals, 72% of the cases were estimated within the error of +/-1.0 h and 92% within +/-5.0 h. Although assessments of non-temperature based PM changes are mostly subjective and have a wide variation, the present study demonstrated a usefulness of non-temperature based PM changes in the estimation of PM intervals.

HPLC analysis of anti-malaria agent, chloroquine in blood and tissue from forensic autopsy cases in Tanzania.

HPLC analysis of anti-malaria agent, chloroquine (CQ) in blood and tissues with a simple HCl back extraction method was applied to three forensic autopsy cases in Dar es Salaam, Tanzania. CQ concentrations in femoral vein blood were 8.5, 48.4 and 43.8 microg/ml in three cases, respectively, which were high enough to attribute the cause of deaths to an acute CQ poisoning. There were great site dependent variations in blood CQ levels. The right heart blood samples were very high, which may be explained by incomplete distribution of the drug before death or postmortem diffusion from liver and its surrounding blood, as high CQ levels were remarkable in the liver. Suicidal and accidental CQ poisonings are very common and CQ is a very important chemical in the field of forensic toxicology in Tanzania.

Fatal intussusception in infancy: an experience in forensic autopsy.

Intussusception, although a common cause of pediatric surgical emergencies, is a rarely fatal condition. A 7-month-old infant who was discovered in her cot was unresponsive and pronounced dead after 2 h of uneventful cardiopulmonary resuscitation in an emergency hospital. Forensic autopsy which was performed in order to clarify the circumstances surrounding the death revealed intussusceptions at two sites of the ileum. Although morbidity and mortality rates from the condition have progressively declined in recent decades but avoidable deaths still occur as was experienced in the present case. The forensic pathology significance in this case was the occurrence of 'painless intussusception' whereby the affected child clinically exhibited no discomfort or characteristic features of acute abdomen until death. In summary, the present case has exhibited an uncommon fatal occurrence and demonstrated the importance of forensic autopsy in such unexpected sudden infant deaths.

Inhibitory activity of epigallocatechin gallate (EGCg) in paraquat-induced microsomal lipid peroxidation--a mechanism of protective effects of EGCg against paraquat toxicity.

Recently we have reported that epigallocatechin gallate (EGCg), a major component of Japanese green tea, significantly increased the survival rate of paraquat (Pq) poisoned mice. This paper describes two biochemical activities of EGCg, which relate to its protective effects against Pq toxicity. EGCg inhibited Pq-induced microsomal malondialdehyde (MDA) productions in rat liver microsome system containing 40 microM FeSO(4). Forty micromolar EGCg inhibited MDA production significantly. EGCg may inhibit the Pq-induced MDA production by at least two mechanisms. One may be iron-chelating activity as the inhibition disappeared when excess amounts of FeSO(4) were added to the reaction mixture, which indicated that EGCg reduced iron driven lipid peroxidation by pulling out available irons in the reaction mixture. The other is radical scavenging activity. EGCg scavenged DMPO-OOH spin adducts generated by the microsome-Pq system. The dose response curve of EGCg was similar to that obtained by ascorbic acid which is a typical water-soluble radical scavenger. Although ascorbic acid had a potential activity of scavenging superoxide radicals, it can not be recommended to use for the treatment of Pq poisoning, because ascorbic acid acts as a pro-oxidant in the presence of free transition metal ions by accelerating the Fenton reaction (Fe(2+)+H(2)O(2)-->Fe(3+)+OH(-)+OH*), which is responsible for lipid peroxidation. On the contrary, EGCg inhibited iron-driven lipid peroxidation presumably not only by chelating to Fe ions but also by scavenging superoxide radicals, which are responsible for the reduction of ferric (Fe(3+)) to ferrous (Fe(2+)) that catalyzes the Fenton reaction. Chelating and radical scavenging activity of EGCg can be expected simultaneously in the occurrence of Pq toxicity, which may explain the protective effects of EGCg against Pq toxicity.

[Ethanol concentrations in multi-site sampling blood in forensic autopsy cases--a retrospective analysis over a period of six years (1994-1999) in Kumamoto University].

Ethanol and n-propanol concentrations in forensic autopsy cases determined in Department of Forensic Medicine, Kumamoto University School of Medicine were reviewed retrospectively. Out of 388 autopsies in 6 years (1994-1999), ethanol was positive in 88 (22.7%) cases. Higher positive rates were observed in bleeding and burning cases compared to other cases. Histograms of the blood ethanol concentrations in all ethanol positive cases had two peaks at 0.1 mg/ml to 0.5 mg/ml and 1.5 mg/ml to 2.0 mg/ml ranges, which indicated that not only an intermediate but also a weak drunkenness level could be a risk factor of being involved in forensic fatalities. There were no differences in mean ethanol concentrations in the blood samples of the right, left and whole heart blood collected from each victim. The femoral blood, however, was slightly higher than those of heart blood. N-Propanol, an indicator for postmortem ethanol production, was detected in 14.7% of stomach contents samples as early as 6 to 12 hours of post mortem intervals, whereas it was not remarkable in urine and femoral vein blood.

A simple dot enzyme-linked immunosorbent assay for ABO blood typing of biological fluid and stains: effects of heating samples.

A simple dot enzyme-linked immunosorbent assay (Dot-ELISA) using commercially available monoclonal anti-A and anti-B antibodies and biotinylated anti-H lectin was developed for ABO blood typing of biological fluid and stains. Its application to forensic practice was examined with 117 saliva samples and their stains, and practical case samples of 8 seminal, 6 vaginal and 45 aged salivary stains. In the simple Dot-ELISA, a new step to heat biological samples was introduced in the system in order to block unfavorable non-specific reactions of the samples with secondary enzyme conjugate. The simple Dot-ELISA could determine accurately the ABO blood type of a small amount of secretor's and non-secretor's salivary samples. In practical tests of seminal, vaginal and salivary stains, all results were confirmed to be identical to those determined by the conventional absorption-inhibition test and the absorption-elution test. The simple Dot-ELISA is considered to be accurate, rapid, simple, sensitive and easy to perform in routine forensic practice. It is also a unique and helpful method to determine the ABO blood types of various biological samples.

Neuropathological diagnosis of Alzheimer's disease in forensic autopsy of elderly persons with fatal accident.

Cognitive dysfunction in Alzheimer's disease may lead to accidental deaths in the elderly. Neuropathological diagnosis of the disease is, therefore, an important issue in forensic autopsy to determine the causal relation to accidents. To evaluate the suitability of the current histopathological diagnostic criteria for Alzheimer's disease by Khachaturian and Mirra et al. in elderly persons dying from accidents and coming for forensic autopsy, we studied the brains of nine demented and 12 non-demented persons by silver stain and immunohistochemistry. When the density of senile plaque was applied to the criteria, only four out of nine demented persons met the criteria for definite Alzheimer's disease. The demented persons had significantly higher density of diffuse plaque and higher frequencies of amyloid angiopathy, neurofibrillary tangle and neuropil thread than the non-demented persons. These results indicated that the current diagnostic criteria do not always diagnose Alzheimer's disease in forensic autopsy of elderly persons with fatal accident. The presence of abundant diffuse plaque, neurofibrillary tangle, amyloid angiopathy and neuropil thread may help to diagnose Alzheimer's disease in forensic autopsy.