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Evidence for the presence of m-tyramine, p-tyramine, tryptamine, and phenylethylamine in the rat brain and several areas of the human brain.

Postmortem human brains have been obtained from four nonpsychiatric patients, aged 59-70 years. Regional analysis of the trace amines phenylethylamine, p-tyramine, m-tyramine, and tryptamine has indicated that the amines are distributed heterogeneously throughout the brain, but are most concentrated in the basal ganglia. Although the levels are very low, evidence obtained from animal studies has indicated that the trace amines have a very rapid turnover rate. Their presence in a brain synaptosomal fraction suggests a possible involvement in the process of neurotransmission. Postmortem changes in human brain amines are discussed in relation to those occurring postmortem in the rat brain, in which phenylethylamine, p-tyramine, and tryptamine have been shown to increase to levels greater than those prevailing in vivo.

Aged↗

Epidural herniation of the cerebral tissue in a burned body: a case report.

A case in which herniation of the cerebral tissue was seen in the epidural space of a burned body is reported. The dead, burned body of an 8-year-old female was found in her burnt-out house. Externally, the body (except the back) was burned to the third or fourth degree, although there was no injury, specifically no skull fracture due to a mechanical force. Internally, much soot deposit was observed in the trachea, main bronchi and peripheral bronchioles. A 9.5 cm x 6.0 cm, thermo-coagulated, yellowish-white mass was unexpectedly found lying on the dura mater of the left temporal lobe. Histologically, H&E stain showed that the mass was thermo-coagulated cerebral tissue, although neither intra-cerebral haemorrhage nor cerebral contusion was found. The carboxyhaemoglobin (CO-Hb) concentration in the intracardiac blood was 83%. The cause of her death was diagnosed as death due to fire, based on the autopsy findings described above and the high CO-Hb concentration value. Therefore, the unexpected entry of the cerebral tissue into the epidural space was considered to be one of the morphological changes due to the local action of heat; the authors propose that such a postmortem change should be called 'heat brain herniation'.

Brain Injuries↗

Dynamic high-resolution 1H and 31P NMR spectroscopy and 1H T2 measurements in postmortem rabbit muscles using slow magic angle spinning.

Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07).

Animals↗

Drugs in postmortem adipose tissues: evidence of antemortem deposition.

INTRODUCTION: Drug concentration measured in postmortem adipose tissue may or may not reflect antemortem concentration. To examine the possibility of whether the presence of basic drugs in adipose tissue is the result of postmortem change, we examined: tissues with and without livor mortis, concentration gradients within the adipose layer, and the stability of drug concentrations during the postmortem period. CASE REPORTS: Five drug-related deaths with case histories and analytical data are presented. Adipose tissues with and without livor mortis from the thigh area of the same decedent were analyzed for cocaine. The cocaine concentration of the tissue exhibiting 4+ livor was equivalent to the concentration observed in tissue without livor. Analyses of cross sections of adipose tissues containing cocaine and methamphetamine disclosed that drug concentrations were equally distributed throughout the layer, from just beneath the dermis to directly above the muscle. When morphine and temazepam concentrations were measured in adipose tissues collected from similar sites, but at different times, from the same cadaver, they remained essentially the same over 3 days (approximately 80 h). CONCLUSIONS: Since concentrations were the same in areas with and without livor mortis, the possibility of redistribution into adipose from blood or vascular channels is eliminated. The absence of a concentration gradient within the adipose layer rules out diffusion or permeation from muscle into the adipose layer, and the failure of morphine or temazepam concentration to change over time indicates that drugs in the adipose tissue are stable during the postmortem interval. Our findings support the notion that drugs identified in postmortem adipose tissue are there because of antemortem deposition and not because of any postmortem change or event.

Adipose Tissue↗

Freeze-replica study of lipid droplets in the Ito cells of rat liver.

The structure of lipid droplets in the Ito cells was studied by the freeze-replica method in vitamin A-treated rats. Most lipid droplets were fractured in planes through their lipid contents. Some of them were surrounded by a membrane structure that had intramembranous particles (IMP), which were more numerous on the protoplasmic face than the exoplasmic face. Those with the membrane containing IMP correspond to the membrane-bounded lipid droplets (lipid droplets surrounded by lysosomes) seen in the electron microscopic observation of conventional thin section preparations. There were no lipid droplets showing concave or convex multilayered structures in the Ito cells from properly fixed livers. Since these multilayered structures were observed only in the materials fixed by immersion with glutaraldehyde or fixed 40 minutes after the death of animals, it is likely that they represent an artifact, showing an autolytic change, caused by poor fixation or postmortem change.

Animals↗

Early-phase postmortem redistribution of the enantiomers of citalopram and its demethylated metabolites in rats.

The aim of this study was to investigate the early-phase postmortem redistribution of the enantiomers of citalopram (CIT) and its metabolites demethylcitalopram (DCIT) and didemethylcitalopram (DDCIT) in a rat model. Furthermore, we wanted to examine the role of the lungs as a reservoir of postmortem drug release and to investigate the influence of storage temperature (21 degrees C vs. 4 degrees C) on postmortem changes. Rats were administered a single CIT dose of 100 mg/kg (s.c.), and heart blood and lung samples were collected antemortem and 15 min postmortem for enantioselective high-performance liquid chromatographic analysis. About three times higher blood drug and metabolite levels were observed in the postmortem rats than in the antemortem rats (p < 0.0001). Refrigeration at 4 degrees C did not prevent, but significantly reduced, the postmortem increase in heart blood CIT levels as compared to the concentrations in the rats stored at 21 degrees C (p < 0.05). The lung drug concentrations were lower postmortem than antemortem (p < 0.05). The enantiomeric (S/R) concentration ratios of CIT and metabolites in blood and lungs were of similar magnitude before and after death. The parent-drug-to-metabolite ratios for CIT/DCIT were unchanged after death. In conclusion, this study shows that heart blood CIT and metabolite levels increase rapidly after death. Further, a fall in postmortem CIT concentrations in the lungs was observed, indicating that the lungs seemed to represent one major source of drug release during early-phase postmortem redistribution.

Animals↗

Lipid content in the liver of fatty metamorphosis of pregnancy.

Lipid analyses were performed on the liver of a patient who died during an episode of acute fatty liver of pregnancy, and on livers from normal subjects and from subjects suffering from nutritional fatty livers. Comparison of these data indicates that in fatty liver of pregnancy the increased hepatic lipids consist primarily of free fatty acids. The recognized toxicity of fatty acids suggests a pathogenic mechanism for the disease. Nutritional fatty liver is associated predominantly with an increase in triglyceride. These changes are not the result of postmortem change, and they confirm and extend the previous data concerning the fat accumulation in human hepatic illness.

Adult↗

Time-dependent expression of interleukin-10 (IL-10) mRNA during the early phase of skin wound healing as a possible indicator of wound vitality.

This study was performed to clarify whether interleukin-10 messenger RNA (IL-10 mRNA) could be a possible indicator for the distinction between intravital wounds and postmortem damage. After incision, mice were sacrificed from 0 to 180 min. The initial amount of IL-10 mRNA in each skin specimen was evaluated using the reverse transcriptase-polymerase chain reaction (RT-PCR). After 15 min there was a rapid increase in IL-10 mRNA which peaked at 60 min. A significant increase in IL-10 mRNA occurred between 30 and 180 min. During the 5 day postmortem interval the increase in time-dependent IL-10 mRNA expression was maintained and no significant increase in IL-10 mRNA expression occurred in the postmortem control. The increased expression of IL-10 mRNA could be considered a vital reaction in skin specimens with postmortem change. This study demonstrated the possible use of mRNA analysis for forensic wound examination because mRNA was detectable by RT-PCR over a longer postmortem time course.

Animals↗

[The use of impedance plethysmography in the forensic medical diagnosis of the time of death].

Diagnosis of the time of death (DTD) is a pressing problem of forensic medicine. For defining the criteria of DTD by impedance plethysmography, electric conductivity of tissues was studied in 45 corpses of male subjects aged 18-44 years who died from craniocerebral injuries. Autolytic changes were characterized by a wave-like pattern, with periods of stability replaced by necrobiotic processes. This allows recording residual activity of surviving cells and tissues in a corpse during the early postmortem period. Based on these data, a model of postmortem changes in tissues can be created for DTD.

Adult↗

Postmortem locus coeruleus neuron count in three American veterans with probable or possible war-related PTSD.

The authors investigated whether war-related posttraumatic stress disorder (WR-PTSD) is associated with a postmortem change in neuronal counts in the locus coeruleus (LC) since enhanced central nervous system (CNS) noradrenergic postsynaptic responsiveness has been previously shown to contribute to PTSD pathophysiology. Using postmortem neuromorphometry, the number of neurons in the right LC in seven deceased elderly male veterans was counted. Three veterans were classified as cases of probable or possible WR-PTSD. All three veterans with probable or possible WR-PTSD were found to have substantially lower LC neuronal counts compared to four comparison subjects (three nonpsychiatric veterans and one veteran with alcohol dependence and delirium tremens). To the authors' knowledge, this case series is the first report of LC neuronal counts in patients with PTSD or any other DSM-IV-TR anxiety disorder. Previous postmortem brain tissue studies of Alzheimer's Disease (AD) demonstrated an upregulation of NE biosynthetic capacity in surviving LC neurons. The finding reported is consistent with the similar upregulation of NE biosynthetic capacity of surviving LC neurons in veterans who developed WR-PTSD. Especially if replicated, this finding in WR-PTSD may provide further explanation of the dramatic effectiveness of propranolol and prazosin for the secondary prevention and treatment of PTSD, respectively. The LC neurons examined in this study are probably the origin of the first or second "leg" of what might be termed the PTSD candidate circuit. Larger neuromorphometric studies of the LC in veterans with WR-PTSD and in other development-stress-induced and fear-circuitry disorders are warranted, especially using VA registries.

Aged↗

Basal level of prostaglandin D2 in rat brain by a solid-phase enzyme immunoassay.

A solid-phase enzyme immunoassay for prostaglandin D2 (PGD2) was developed in which PGD2 was labeled with horseradish peroxidase. After competitive binding to the immobilized antibody between enzyme-labeled and free PGD2, the activity of the enzyme bound to the antibody was assayed fluorometrically using 3-(p-hydroxyphenyl)-propionic acid and hydrogen peroxide as substrates. The procedure allowed determinations of 3-100 pg for PGD2. The IC50 value for PGD2 in the solid-phase enzyme immunoassay was about 25 pg and the sensitivity was improved about 10 times compared to those in radioimmunoassay and in solution-phase enzyme immunoassay. The solid-phase enzyme immunoassay was applied to the measurement of PGD2 content in rat brain and thereby an octadecylsilyl silica cartridge and a reversed-phase HPLC were sequentially used for sample preparations. Heads were immediately frozen in liquid nitrogen after decapitation to avoid a postmortem formation of PGD2. PGD2 contents measured by solid-phase enzyme immunoassay correlated well with the values obtained by radioimmunoassay (r = 0.966) after raising its contents by intravenous administration of PGD2. The in vivo level of PGD2 in rat brain was extremely low but determined to be 0.11 +/- 0.03 ng/g tissue (mean +/- S.E.M.) with this enzyme immunoassay. The result was equal to the value extrapolated to zero time from the postmortem change.

Animals↗

Application of labeling techniques to archival temporal bone sections.

Pathology of the human inner ear has traditionally been studied in celloidin-embedded, hematoxylin and eosin-stained sections of the temporal bone. Although the traditional histologic approach has yielded valuable information, it is now possible to extend these studies to include analysis of molecules using immunohistochemical and histochemical staining techniques. Fourteen antibodies and 6 lectins have been applied to 420 archival, celloidin-embedded human temporal bone sections. Tissues had been fixed in 10% formalin, embedded in celloidin, sectioned, and stored for as many as 40 years. The staining intensities varied among sections, so they were ranked from 'no label" to "dense label." To investigate the relationships between the extent of postmortem changes (PMCs), storage time, and staining intensity for each antibody, the sections were graded according to their PMCs, which ranged from good preservation of the temporal bone histologic structure to severe postmortem autolysis. Although statistical analysis indicated that both extent of PMCs and storage time in general decrease the staining intensity, both poorly fixed tissue and sections stored for a long time can yield good immunostaining results with some antibodies.

Antibodies↗

Cardiac blood pH as a possible indicator of postmortem interval.

Postmortem changes in the pH of blood and selected tissues in rats were evaluated at intervals ranging from 2 min to 96 h. Cardiac blood pH was significantly and reproducibly decreased in all groups at all postmortem intervals, independent of the method of sacrifice used. A preliminary study using cardiac blood obtained at autopsy from a limited number (n = 11) of human subjects demonstrated a significant negative correlation (r = -0.908, P less than 0.01) between postmortem interval (range 2 to 20 h) and cardiac blood pH.

Adult↗

The use of vitreous humor as an alternative to whole blood for the analysis of benzodiazepines.

In postmortem drug analysis, the most commonly used sample matrix is whole blood. However, postmortem changes can denature this matrix, resulting in a loss or degradation of drugs, thus biasing analytical findings. Vitreous humor is thought to be less affected by these changes and should, therefore, have the potential to provide a more reliable estimation of antemortem drug concentrations. To assess the usefulness of vitreous humor for the analysis of benzodiazepine drugs, vitreous humor and whole blood were obtained postmortem in 27 cases. Three benzodiazepine drugs were investigated-temazepam, diazepam, and desmethyldiazepam. For temazepam and diazepam, some correlation was found between the matrices (R2 = 0.789 and 0.724, respectively). However, for desmethyldiazepam, no correlation was observed (R2 = 0.068). Regression analysis on plots of vitreous humor versus blood concentrations produced gradients of less than 1.0 showing that, in general, levels in blood are higher than the corresponding levels in vitreous humor.

Benzodiazepines↗

Canid modification of human remains: implications for time-since-death estimations.

Time-since-death estimations are usually based on physical decomposition of the corpse, insect succession, and contextual associations. The rates of change and succession are based on decomposition studies, most of which control access of scavengers to the corpse; however, many naturally exposed corpses are subject to scavenger modification. These modifications change the rate of decomposition, the pattern of insect succession, and the context of associations, thus altering estimations of time since death. A controlled feeding study with captive wolves and road-killed deer is pertinent to understanding canid scavenging and how scavenging may alter postmortem changes. During feeding, the wolves commonly dismember and devour the deer in a predictable sequence. Although there are some variations in the usual sequence, the carcass is always moved, and skeletal elements are separated, diminished in size and scattered. Scavenging must, therefore, be considered in estimating time since death.

Animals↗

The remains of Dr. Carl Austin Weiss: anthropological analysis.

Anthropological analysis of the remains of Dr. Carl Austin Weiss revealed biological information consistent with his known characteristics. Postmortem changes included decomposition of amalgam dental restorations, likely releasing mercury that stained the anterior dentition, deposition of sulphur compounds on much of the skeleton, and the formation of numerous small bone fractures. Analysis revealed extensive perimortem trauma, indicating multiple gunshot wounds to the upper body. Trajectory analysis of the bony trauma suggested that at least 20 projectiles penetrated Dr. Weiss's body. They originated from many different directions, but mostly from the posterior. No bony changes associated with perimortem trauma of the hands were noted.

Bone and Bones↗

Cyanide dermatosis or a post-mortem artefact?--A case report.

A case of a chronic alcoholic, found dead in the afternoon after consuming an intoxicant early in the morning and showing post-mortem skin slippage with a blackened charred-like appearance, is reported. Viscera chemical analysis gave a positive test for significant quantities of cyanide. However, on histopathological examination, since no evidence of blister formation or vital reaction in the skin tissues were found, this was thought to be a very short duration, accelerated postmortem change phenomenon, due to local heating from contact with the sun-baked terrace.

Adult↗

[Experimental studies of postmortem diffusion of toluene in blood, brain, muscle and fat of rats exposed to toluene vapor].

This report describes the postmortem diffusion of toluene in blood, brain, muscle and fat of rats exposed to toluene vapor. After exposure to toluene vapor (4,000 ppm) for 15 and 60 min, rats were killed by CO2 exposure and left for 12 and 24 hours in a fresh air. Toluene concentrations in blood, brain, muscle and fat were determined by head space gas chromatography. Postmortem decrease of toluene concentration in blood and increase of that in fat were observed with postmortem interval in both exposure conditions. But an extremely larger postmortem changes in the short exposure than those in the long exposure were observed. These are due to the facts that the value of fat/blood toluene concentration ratio at the time of death in the short exposure (3.3) was extremely smaller than that in the long exposure (10.5). Postmortem increase in muscle was also observed slightly. Postmortem concentrations in brain in both exposure intervals were relatively constant during these experiments. This postmortem stability is attributable to its anatomical features. These results suggest that brain toluene concentration at the autopsy is useful for the estimation of blood toluene concentration at the time of death and by comparison between toluene concentration in blood and that in brain it may be possible to presume the circumstances of death.

Adipose Tissue↗