Postmortem Injuries Research Articles

Distinguishing antemortem from postmortem injuries by LTB 4 quantification

Leukotriene B 4 (LTB 4) in skin samples from seven forensic cases was detected by HPLC to distinguish their antemortem or postmortem origin. In total, there were thirteen antemortem and seven postmortem specimens. The results showed that LTB 4 was found in all antemortem wound specimens which were either fresh, or refrigerated or fixed in formalin for less than 10 days. In contrast, LTB 4 could not be detected in postmortem wound specimens. These results suggested that detecting the content of LTB 4 is a useful method for distinguishing antemortem from postmortem injuries.

Diagnostic value of ions as markers for differentiating antemortem from postmortem wounds

In the present study, the diagnostic value of ions as markers for differentiating antemortem from postmortem wounds was investigated. To accomplish this, skin and muscle samples were taken from antemortem and postmortem wounded cadavers and the contents of ions (Fe, Zn, Mg, Cu, K and Na) were determined with atomic absorption spectrophotometery. Our results show that the concentrations of Fe in antemortem wounded skin and muscle were significantly higher as compared with the control skin and muscle and the K Na ratio in antemortem wounded muscle was reduced significantly compared to control muscle. However, the concentrations of Zn, Mg, Cu in antemortem wounded skin and muscle, and the K Na ratio in antemortem wounded skin, were not significantly different from their controls. The concentrations of all these ions in postmortem wounded skin and muscles were not significantly different from their controls. It is indicated that the concentrations of Fe in skin and muscle and K Na ratio in muscle are useful markers for differentiating antemortem from postmortem wounds. However, their routine use in forensic practice needs further investigation, since there are some overlaps of the ionic concentrations between normal and wounded skin and muscle.

Immunohistochemical analysis of markers for different macrophage phenotypes and their use for a forensic wound age estimation.

A total of 117 vital skin wounds (post infliction intervals between a few seconds and 7 months), 20 postmortem wounds and skin specimens with beginning or advanced signs of putrefaction were investigated. Different markers for macrophage maturation (27 E 10, RM 3/1, 25 F 9, G 16/1) were analyzed by immunohistochemistry. The early stage inflammation marker 27 E 10 stained macrophages, but also monocytes and neutrophilic granulocytes localized in blood vessels or bleeding induced postmortem and therefore provided no further information for a forensic wound age estimation in comparison to the routine histological detection of macrophages. The antigens recognized by the RM 3/1- (intermediate stage inflammation marker) and 25 F 9-antibodies (late stage inflammation marker) were expressed exclusively by histiocytes and inflammatory cells that had migrated from the blood vessels as part of the acute inflammatory response associated with an intravital reaction. The morphometrical analysis revealed positive results (defined as at least a two-fold increase in number in 2 or more microscope fields when compared to the maximum value of histiocytes found in uninjured skin) for the RM 3/1- or 25 F 9-antibody earliest in wounds aged 7 or 11 days, respectively. Similarly to the 25 F 9-antibody, the chronic stage inflammation marker (G 16/1) reacted with a macrophage subpopulation first detectable 12 days after wounding but showed positive results in a comparably reduced percentage of cases. On the other hand, this marker did not stain a relevant number of resident macrophages thus facilitating the evaluation of the specimens.(ABSTRACT TRUNCATED AT 250 WORDS)

A preliminary study of skin electrical injury with computerized image analysis

This paper reports the application of computerized image analysis to a quantitative study of skin electrical injury in order to provide an objective and quantitative standard for identifying the electrical mark. The differences between antemortem and postmortem electrical injuries were also studied. In normal and electrically-injured human skin, the long/short axes ratio of nuclei were measured. The mean of epithelial basal cells was 1.4344 and 5.9325, respectively, the mean of hair follicle cells was 1.6193 and 4.5988, respectively, and the mean of capillary endothelial cells was 4.0229 and 4.3593, respectively. In rat specimens, the long/short axes ratio of nuclei of normal skin, at 15 and 5 min ante-mortem and at 5 min postmortem injury, the mean epithelial basal cells was 1.4124, 4.5417, 5.2282 and 5.0447, respectively. The mean of hair follicle cells was 1.4140, 4.4863, 4.1146 and 4.4522, respectively, while the mean of capillary endothelial cells was 2.8398, 3.7514, 3.6159 and 3.6977, respectively. The results indicate that the differences between electrical injuries and normal skin with respect to the ratios of long/short axes of the epithelial basal cell nuclei and hair follicle cell nuclei are of remarkable significance. On the other hand, no significant difference could be found between antemortem and postmortem injuries, nor between electrical injury and normal skin, concerning the long/short axes ratios of capillary endothelial cell nuclei. This result provides a preliminary objective standard for the changes of electrical skin injury.

Postmortem injuries inflicted by domestic golden hamster: morphological aspects and evidence by DNA typing

A case of postmortem animal scavenging by a domestic golden hamster ( Mesocricetus auratus) is presented. A 43-year-old woman, who was not under medical treatment, was found dead in her flat with the lower part of her body naked and her legs straddled. Her face showed extensive lesions of the soft tissues which the medical examiner interpreted as vital scalping injuries. The total findings at the scene suggested at first a sexual offence. On autopsy the face injuries were identified as postmortem defects by animal scavenging with the teeth marks typical of rodents. In fact, the deceased had kept in her flat a free-range golden hamster whose burrow contained numerous fingernail-sized pieces of skin, fatty and muscular tissue. By means of DNA typing it was proved that these pieces of tissue belonged to the deceased.

Postmortem Injuries by Indoor Pets

Four cases of postmortem injuries caused by indoor pets (three by dogs and one by cats) are presented. A pattern which is associated with this phenomenon is described. The important common factors appear to be the presence of free-moving pets inside the house, social isolation of the deceased, and the victim having a predisposing condition causing sudden death.

Estimation of local vital reactions in severely burned tissues of guinea pig skin using C3a or C3a desArg as a marker

We observed changes in levels of anaphylatoxin C3a and/or its desArg (C3a/C3a desArg) peptides in the local site of severely burned skin tissues of guinea pigs using immuno-Western blotting methods. The C3a/C3a desArg peptides, which were probably generated during complement activation, were detected at significant levels from 30 min to 72 h following burn injury in an area limited to 3 mm from the wound edge. Levels of these peptides showed a tendency to be highest in that area 24 h after burn infliction. In postmortem injuries, C3a/C3a desArg peptides could not be detected. These peptides were detected in antemortem wounds up to at least 2 days at 22 degrees C and up to 3 days at 4 degrees C after death, although decreases in levels were found. Lower concentrations of these peptides were also found in postmortem burns in which postmortem hypostasis appeared strongly. These results suggest that, except for injuries of the area with obvious postmortem hypostasis, detection of C3a/C3a desArg can be useful for estimation of vital reactions in many kinds of wounds during inflammation.

Complement component C3a or C3a desArg as a new marker for estimation of local vital reactions in incised skin wounds

The anaphylatoxin C3a or its desArg form ( C3a desArg ) generated during complement activation could be detected in the vicinity of incised skin wounds of guinea pigs using immunoblotting methods. The C3a desArg peptides were detectable immediately after injury in local sites up to 3 mm from the wound edge. In subsequent determinations of up to at least 3-day-old antemortem wounds, the maximum concentration of these peptides was largely localized up to 6 mm from the wound edge at 2 h after injury. In postmortem wounds, however, these peptides were undetectable. When they were released in antemortem wounded tissues they could be detected up to 1 day at 22°C after death. These results suggest that the detection of C3a desArg in wounds using immunoblotting methods can be useful for distinguishing ante- from postmortem wounds.

Altered collagen (fibrinoid change) at the site of post-mortem injuries.

The terms fibrinoid degeneration and fibrinoid substance were first introduced by Neumann in 1880 to describe some alterations in the staining characteristics of collagen. Collagen fibres which have undergone this change are eosinophilic, homogeneous and take some of the tinctorial properties of fibrin. Lendrum et al. (1962) developed Martius-Scarlet-Blue (MSB) stain, which is preferentially taken up by fibrin. The principle of the method described by Lendrum and his colleagues is the use of acid dyes of different molecular size (Martius yellow, Brilliant Crystal Scarlet 6R, methyl blue) in accordance with the alteration in structure of fibrin at different stages of development. The newest fibrin likely to be found in sections is presumably the fine network of postmortem fibrin. Much of this takes a yellow stain with the MSB method, as do erythrocytes. Slightly older fibrin (16 hours) takes a bright red stain with the MSB. Complete validity of the method depends on prolonged fixation in formal-sublimate. It gives brilliant staining with acid dyes and enhances metachromasia. In the Department of Forensic Medicine in Leeds it was noticed that an alteration in collagen staining with MSB stain was taking place in samples collected from different kinds of injuries, e.g. ligature marks, electrical injuries and burns. Similar changes were only occasionally seen in abrasions. It was therefore decided to inflict different kinds of injuries on the skins of rats and to fresh pieces of human skin in vitro, to observe any alterations in collagen staining with MSB and to assess the significance of these changes in relation to the timing of injuries.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization and quantification of the nonspecific esterase in injured skin for timing of wounds

Histochemical quantification of the nonspecific esterase (NSE) in injured skin was performed using histochemical demonstration of the enzyme and a microspectrophotometric scanning technique on specimens taken from 32 Hartley guinea-pigs and 8 cases of human skin wounds. In all antemortem incisions and lacerations, including those made at the agonal state, NSE activity could be observed in the dermal tissue of the wound edge. The enzyme activity increases with the antemortem duration of the injuries. Both total content and mean concentration of NSE in the wound edge between antemortem and postmortem wound groups differ significantly (< 0.01). Multiple range test shows that significant differences ( P < 0.05 or P < 0.01) of total content of NSE in the wound edge also exist in 0 – 5 min, 15 – 30 min, 1-h, 2-h and 4-h antemortem incised wound groups and in 0-min, 5-min, 15 – 30 min, 1-h and 4-h antemortem laceration groups. The positive NSE reactions in 8 cases of human skin wounds were similar. The study indicates that histochemical quantification of NSE in injured skin is very useful in timing wounds and is exactly applicable in medicolegal practice. According to the different influences of inhibitors on enzymes, it was inferred that the enzyme activity in wound edges was due to B-esterase.

Immunohistochemical localization of fibronectin as a tool for the age determination of human skin wounds.

We analyzed the distribution of fibronectin in routinely embedded tissue specimens from 53 skin wounds and 6 postmortem wounds. In postmortem wounds a faint but focal positive staining was exclusively found at the margin of the specimens which did not extend into the adjacent stroma. Vital wounds were classified into 3 groups. The first comprising lesions with wound ages ranging from a few seconds to 30 min, the second comprising those with wound ages up to 3 weeks, and the third group with lesions more than 3 weeks old. Ten out of 17 lesions with a wound age up to 30 min showed a clear positive reaction within the wound area. Three specimens in this group were completely negative, while in 4 additional cases the result was not significantly different from postmortem lesions. These 7 cases were characterized by acute death with extremely short survival times (only seconds). In wounds up to 3 weeks old fibronectin formed a distinct network containing an increasing number of inflammatory cells corresponding to the wound age. In 2 cases with a survival time of 17 days and in all wounds older than 3 weeks fibronectin was restricted to the surface of fibroblasts and to parallel arranged fibers in the granulation tissue without any network structures. We present evidence that fibronectin is a useful marker for vital wounds with a survival time of more than a few minutes. Fibronectin appears before neutrophilic granulocytes migrate into the wound area.(ABSTRACT TRUNCATED AT 250 WORDS)

Wound documentation in coronial autopsies

Considering that a wound is defined as a disruption of continuity of a tissue by physical injury, it is obvious that for a pathologist involved in Colonial work, defining wounds is part of daily practice. It is a fundamental part of the subject, but as each pathologist feels that they have the correct approach, individual differences can lead to confusion. Wound documentation is of relevance in the majority of Coronial autopsy cases, no more so than in homicide cases. This is because of the sometimes very detailed court room discussions relating to each injury, and the necessity for all persons present to understand fully the nature of the injury. The degree of documentation should be adequate to allow a second pathologist to be put in the shoes of the first pathologist. The autopsy report is discussed in terms of injury definition, style, and nomenclature including differentiation of antemortem from postmortem injuries, as well as complex injuries. The use of charts, diagrams, tracings, photography, video, and X-rays are each considered.

Localization and quantification of histamine in injured skin as parameters for the timing of wounds

Localization and estimation of the histamine (HA) content in skin wound edges in 86 Sprague-Dawley rats and three cases of human injuries were carried out by a microfluorimetric method specific for this amine which forms a complex with o-phthalaldehyde (OPT). Distribution and density of the mast cells in the same areas were observed at the same time by toluidine blue stain. In all skin specimens with antemortem wounds, both the epidermis and upper dermis exhibit extracellular yellowish fluorescence of the HA-OPT complex. The fluorescent zone spreads in the wound edges with the lapse of time in vital injuries. The HA content increases gradually up to 30 min and then the yellow histamine fluorescence in areas 0–200 μm from the wound edge decreases. None of these features can be observed in normal skin and postmortem-injured skin. Mast cell degranulation can be demonstrated in all antemortem-injured skin. No statistical relationship exists between the number of mast cells and the HA-OPT fluorescence in either ante- or postmortem-injured groups. This study indicates that skin HA microfluorimetry by the OPT method is of practical value for distinguishing ante- from the postmortem wounds and for timing antemortem wounds.

Postmortem stability of some markers of intra-vital wounds

We have studied the diagnostic value of several markers of the intra-vital nature of wounds — cathepsin D (EC 3.4.23.5) and ions (Ca, Mg, Cu, Zn and Fe) — after the influence of putrefaction. For this purpose, we have inflicted vital wounds to six pigs, which were killed 20 min later. Ten minutes after death, wounds were excised with 5–6 cm of skin around the incision and maintained at three different temperatures (4, 18 and 28.5 ± 13.4°C). After varying periods of postmortem interval from 0 to 48 h, aliquots of each wound were taken and analyzed with atomic absorption spectrophotometry for ions and with UV-spectrophotometry for cathepsin D. Our results demonstrate that ions conserve their diagnostic ability to differentiate vital from postmortem wounds after the influence of putrefaction. Nevertheless, cathepsin D does not show this ability in these experimental conditions.

The Role of White Blood Cells in Post-Mortem Wounds

Walcher in 1936 pointed out the importance of studying a lesion microscopically in order to distinguish between ante-mortem and post-mortem injuries. According to him and to many other authors (Raekallio, 1984; Ojala et al., 1969; Fatteh, 1966) leucocytic reaction is the earliest histological sign of inflammation, and it is the most reliable sign of the vitality of wounds. The pavementation (also known as margination) of the vascular endothelium of white blood cells, and the beginning of their extravasation, may be seen during the first hour (Ojala et al., 1969), 6 hours (Malik, 1970), 8 hours (Fatteh, 1966) or 8–16 hours (Raekallio, 1964) after the injury.Profuse bleeding and fibrin deposition are no longer regarded as necessarily signs of vitality of wounds. (Shapiro and Robertson, 1962; Laiho, 1967), yet the behaviour of white blood cells in post-mortem wounds remains unknown.It was therefore decided to study the effects of chemotactic materials injected intradermally and in the anterior abdominal wall in dead rats. The experimental findings are presented, along with discussion, and a review of the literature.

Identification from skeletal remains

The remains of a suspected case of homicide, found to be almost totally skeletal on exhumation, was dug out from a pit in a tract of marshy land in the deep south or Sri Lanka. No clothing or other identifying articles were found with the remains. During exhumation a distinct odour of kerosene oil was observed. Examination revealed that the major part of the skeleton was present, including the skull and lower jaw. Examination of these enabled an opinion to be given on the stature, sex and age of the deceased. The head hair found at exhumation showed evidence of burning. Two groups of injuries were found in the bones. One group was consistent with injuries caused during life and showing the characters of healing and moulding. Although medical records were not available, it was possible to obtain a description of the injuries from the court records as a doctor had given evidence on injuries sustained by the deceased years previously. The other group comprised of injuries that would have caused or contributed to death, or were post-mortem injuries inflicted for purposes of disposal. This case illustrates that personal identification may be established from old healed skeletal injuries.

Study of Cathepsin A, B and D activities in the skin wound edges. Its application to the differential diagnosis between vital and postmortem wounds

The aims of the authors in this paper has been to check the diagnostic ability of the Cathepsin A, B and D concentrations in the skin wound edges to the differential diagnosis between vital and postmortem wounds. We have studied 56 domestic pigs grouped in seven experimental series consisting of 8 animals in each, according to the time (0, 5, 15, 30 min and 1, 3 and 6 h) after the enjury. The enzymatic activities were investigated following the methods by Bowen and Davison Biochem. J., 131 (1973) 417–419, Suhar and Marks J. Biochem., 101 (1979) 23–30 and by Anson (modified by Yamamoto, Eur. J. Biochem., 95 (1979) 459–467) for Cathepsin A, B and D, respectively. For the differential diagnosis between vital and postmortem wounds, our results showed that the most useful markers studied are the Cathepsin A and D activities, Cathepsin D that of the first one.

Cathepsin D: a new marker of the vitality of the wound. Comparative study with histamine and serotonin.

We have studied the variation of specific activity of the most important lysosomal enzyme: Cathepsin D. This study has been developed in pig skin incision wounds for its application to the differential diagnosis between vital and postmortem wounds. At the same time, we have determined histamine and serotonine levels to make a comparative study with Cathepsin results. Our results show the utility of this new marker to determine the vitality of wounds, even when the time elapsed until death has been very short. Nevertheless, there is no correlation between Cathepsin D results and those of histamine or serotonin, a fact that induces us to think that this new marker is not useful to determine wound age.

Effect of Dietary Retinyl Acetate, β-Carotene and Retinoic Acid on Wound Healing in Rats

The effects of feeding various levels and combinations of retinyl acetate, β-carotene, or retinoic acid on skin wound healing in rats was investigated. Weanling male Sprague-Dawley rats were fed a vitamin A-free diet for 2 weeks to produce marginal vitamin A status. After a paravertebral incision was made and closed with suture, one of several diets were fed for either 5 or 14 days. Surgery and recovery did not reduce liver vitamin A nor serum retinol levels compared to nonoperated pair-fed controls. Supplemental retinyl acetate feeding at five times the NRC-suggested allowance resulted in a mild, but significantly increased postmortem wound tensile strength after 5 days compared to rats fed the suggested allowance. Although a low level of retinoic acid in the diet (1.3 µg/g diet) depressed wound strength at 5 days, a higher level (5.2 µg/g) increased the strength 57% above controls. Still higher levels (49.1 µg/g) did not further increase the tensile strength of the wound. β-Carotene fed at the requirement level for 5 days (with compensation made for utilization as one-sixth that of retinol) doubled wound strength compared to rats fed the requirement as retinyl acetate. Vitamin A feeding did not enhance wound strength after 14 days of feeding. It is concluded that supplemental retinyl acetate, β-carotene, or in some cases all-trans-retinoic acid can be effective in enhancing wound strength, 5 days but not 14 days after surgery, of young male rats with marginal vitamin A status.vitamin A retinoic acid β-carotene wound healing

Biochemical reconstruction of three cases of death — Results of international cooperation

Biochemical serotonin and histamine determinations were applied to the reconstruction of three suspected homicide cases. To distinguish between ante-mortem and post-mortem wounds and to time the ante-mortem injuries the concentrations of free histamine and serotonin in the wound samples and in the control samples from neighbouring intact skin were examined. The results of these biochemical determinations allowed a reconstruction of the events and one of the three cases was shown to be suicide instead of homicide. These methods can be used at least during the first 4 – 5 days after death and some-times even longer. This allows for international cooperation when investigating and reconstructing complicated cases of death.