The ageing of bruises: a review and study of the colour changes with time.

This work was inspired by a recent case of child abuse where the question of the age of the bruises on the body was raised. The first part of this paper reviews published work on bruises. It illustrates the paucity of work in this field and the absence of studies of the colour changes in bruises of human skin with time. The second part of this paper consists of our own study of the appearance of bruises. The aim was to identify the colour changes which occur in bruises and over what time-scale, in order to determine whether bruises can be aged by appearance. A total of 369 photographs were obtained of bruises aged less than 6 h and up to 21 days old, in 89 subjects aged 10-100 years. It was found that the development of a yellow colour was the most significant change (subjects aged less than 65, P less than 0.001; subjects aged greater than or equal to 65 years, P less than 0.001). The development of a yellow colour occurred significantly faster in subjects aged less than 65 years. (P less than 0.001). The appearance of a blue and purple/black colour was of lesser significance. The appearance of a red colour did not alter significantly with time. From this study it was only possible to conclude that a bruise with a yellow colour was more than 18 h old.     

The ageing of bruises: A review and study of the colour changes with time

This work was inspired by a recent case of child abuse where the question of the age of the bruises on the body was raised. The first part of this paper reviews published work on bruises. It illustrates the paucity of work in this field and the absence of studies of the colour changes in bruises of human skin with time. The second part of this paper consists of our own study of the appearance of bruises. The aim was to identify the colour changes which occur in bruises and over what time-scale, in order to determine whether bruises can be aged by appearance. A total of 369 photographs were obtained of bruises aged <6 h and up to 21 days old, in 89 subjects aged 10–100 years. It was found that the development of a yellow colour was the most significant change (subjects aged <65, P < 0.001; subjects aged ≥65 years, P < 0.001). The development of a yellow colour occurred significantly faster in subjects aged < 65 years. (P < 0.001). The appearance of a blue and purple/black colour was of lesser significance. The appearance of a red colour did not alter significantly with time. From this study it was only possible to conclude that a bruise with a yellow colour was more than 18 h old.     

Supplementary pathway for vitality of wounds and wound age estimation in bruises using the electric impedance spectroscopy technique

Determination of wound vitality and estimation of wound age are central issues in daily forensic practice. The objective of this study was to develop a new and rapid method for determining wound vitality and estimating wound age in bruises using electric impedance spectroscopy. Forty Sprague-Dawley rats (140-170 g) were divided into five groups: group 1 (n=8): controls, group 2 (n=8): postmortem bruises, group 3 (n=8): bruises 1 h before death, group 4 (n=8): bruises 3 h before death, group 5 (n=8): bruises 6 h before death. Measurements of the right gluteus maximus muscle were taken at 6, 24, and 48 h after the rats were sacrificed by cervical dislocation. The results from this study indicate that electric impedance spectroscopy is clearly sensitive enough to differentiate between vital and postmortem wound infliction and to determine the survival time after the infliction of an injury.     

Post-mortem analysis of apoptotic changes associated with human skin bruises

The estimation of the age of skin bruises is of importance in forensic medicine, especially in child abuse cases. Time-dependent changes in bruise colour and/or associated histological features have been used with a limited degree of success. An increased rate of apoptosis in the injured tissue has been considered as a novel time-dependent marker of cell death, by injury inflicted in a rat model. The object of the present study was to apply the TUNEL method of DNA end labelling to identify and enumerate apoptotic cells in bruised and normal skin in order to study the relationship of apoptotic cell density with the age of the bruise. A commercially available DNA end labelling kit, TUNEL method, was standardised, validated and used for this purpose. Twenty unselected post-mortem cases with bruises due to a variety of causes were studied. The apoptotic cells stained with TUNEL reaction were counted in 10 high power fields in the epidermis, as well as in the dermis of formaldehyde-fixed paraffin-embedded skin specimens. The mean positive cell densities (+/- 1 S.E.) were compared with respect to the age of the bruise. In the epidermis, the mean apoptotic cell count was statistically significantly greater in the bruised skin compared to normal skin in 2- to 6-day-old bruises; whilst in the dermis the same was true in 3- to 8-day-old bruises. The overall findings suggest that there is a quiescent period prior to the increase in the apoptotic cell activity that is seen following skin bruising. This is so provided the post-mortem skin samples were collected within a lapse of 6 days or less between the time of death and formalin fixation and paraffin embedding to avoid the bias made by the difference of length of post-mortem interval.     

Complete Cardiac Rupture Associated with Closed Chest Cardiac Massage: Case Report and Review of the Literature

Chest skeletal injuries are the most frequent complications of external chest massage (ECM) during cardiopulmonary resuscitation, but heart and great vessels lacerations that are indeed very rare. We report the case of a 35‐year‐old workman who collapsed and underwent ECM by his co‐workers for almost 30 min. At autopsy, no external injuries, fractures or bruises of the ribs or sternum, were observed. A hemopericardium with a rupture of the heart was found, with no signs of pre‐existent cardiac disease. Bruises of thoracic aortic wall, lung petechiae, a contusion of the liver, and bruises of lumbar muscles were found. The cause of death was due to sudden cardiac death with an extensive cardiac rupture. This is an unusual report of massive heart damage without any skeletal or muscle chest injuries, secondary to cardiopulmonary resuscitation. This kind of cardiac lesions may be considered when thoracic–abdominal trauma, or medical history, is unclear.     

Modification of hematoma findings in the breast region

Subcutaneous bruises caused by blunt injury are common findings in medicolegal case work. If the hematoma involves the anterior thoracic wall, bruises are mostly absent in the region of the nipples and the surrounding areola. A similar phenomenon has already been described for hypostatic skin hemorrhages. A possible explanation for both phenomena is the special tissue texture in the nipple–areola complex. Based on four cases from the forensic autopsy material and two cases from clinical forensic examinations, the macromorphological findings and the histological correlates are presented.     

Ink dating part II: Interpretation of results in a legal perspective

The development of an ink dating method requires an important investment of resources in order to step from the monitoring of ink ageing on paper to the determination of the actual age of a questioned ink entry. This article aimed at developing and evaluating the potential of three interpretation models to date ink entries in a legal perspective: (1) the threshold model comparing analytical results to tabulated values in order to determine the maximal possible age of an ink entry, (2) the trend tests that focusing on the “ageing status” of an ink entry, and (3) the likelihood ratio calculation comparing the probabilities to observe the results under at least two alternative hypotheses. This is the first report showing ink dating interpretation results on a ballpoint be ink reference population.In the first part of this paper three ageing parameters were selected as promising from the population of 25 ink entries aged during 4 to 304 days: the quantity of phenoxyethanol (PE), the difference between the PE quantities contained in a naturally aged sample and an artificially aged sample (R_NORM) and the solvent loss ratio (R%). In the current part, each model was tested using the three selected ageing parameters. Results showed that threshold definition remains a simple model easily applicable in practice, but that the risk of false positive cannot be completely avoided without reducing significantly the feasibility of the ink dating approaches. The trend tests from the literature showed unreliable results and an alternative had to be developed yielding encouraging results. The likelihood ratio calculation introduced a degree of certainty to the ink dating conclusion in comparison to the threshold approach. The proposed model remains quite simple to apply in practice, but should be further developed in order to yield reliable results in practice.     

Alternative light source (polilight) illumination with digital image analysis does not assist in determining the age of bruises

The age of a bruise may be of interest to forensic investigators. Previous research has demonstrated that an alternative light source may assist in the visualisation of faint or non-visible bruises. This project aimed to determine if an alternative light source could be utilised to assist investigators estimate the age of a bruise. Forty braises, sustained from blunt force trauma, were examined from 30 healthy subjects. The age of the bruises ranged from 2 to 231 h (mean = 74.6, median = 69.0). Alternative light source (polilight) illumination at 415 and 450 nm was used. The black and white photographs obtained were assessed using densitometry. A statistical analysis indicated that there was no correlation between time and the mean densitometry values. The alternative light source used in this study was unable to assist in determining the age of a bruise.     

小鼠皮肤创伤愈合过程中IL-10在不同表达部位的组化定量研究

目的探讨小鼠皮肤创伤愈合过程中IL-10在不同表达部位的变化与损伤时间的关系。方法应用免疫组织化学技术和形态计量法,对小鼠不同时程皮肤切创组织中IL-10不同表达部位(表皮细胞及表皮下组织的浸润细胞)的变化进行研究。结果皮肤切创前后表皮层均有阳性表达,切创后1～3h表皮细胞阳性表达水平迅速升高,24h时降至较低水平,伤后48h表达水平再次升高,96h后逐渐回复到正常水平;表皮以下阳性部位主要是浸润的单核/巨噬细胞,损伤6h后阳性细胞在真皮、皮下组织及创口肉芽组织内逐渐增多,于伤后72h达最大值(51.41±3.12)%,96～168h阳性细胞数逐渐减少(29.38±2.64)%～(5.56±4.74)%。结论皮肤创伤修复过程中IL-10在不同表达部位的变化特点与皮肤损伤时间相关,检测其表达变化可望用于损伤时间推断。     

A case of extreme agitation and death after the use of mephedrone in The Netherlands

A 36-year old man, having injured himself severely by smashing windows in a rage of fury, was arrested by the police. He died despite resuscitation attempts. The forensic autopsy showed many superficial skin lacerations, bruises and minor brain swelling, but there was no definitive cause of death. Toxicological analysis showed a high concentration of mephedrone in femoral blood (5.1mg/L) and traces of cocaine, MDMA and oxazepam. The remaining dose of mephedrone in the stomach contents was estimated at 113 mg. Tablets that were found in the house of the deceased also contained mephedrone. We attribute this man's death to a fatal oral intake of mephedrone, which probably led to a state of excited delirium. This was aggravated by blood loss from multiple wounds.     

鼠窒息后缺氧诱导因子1-α在心、肺中的表达

目的研究大鼠机械性窒息死亡后缺氧诱导因子1-α(HIF1-α)在心、肺中的表达变化,探讨其作为窒息死亡诊断指标的可行性。方法制作大鼠缢死模型,分别于窒息死后及正常断颈处死后0、2、6、24h时间段取材,用免疫组织化学、RT-PCR方法测定缢死后HIF1-α在心肌和肺组织中的分布和表达变化。结果HIF1-α在窒息组的心肌和肺组织中表达,窒息组和对照组在各时间段表现出明显差异,对照组仅在死后2、6、24h阳性表达。窒息死亡组可见核阳性表达,而对照组未见核阳性表达。半定量RT-PCR结果显示HIF1-α在0h时各组未见升高,但在2、6、24h窒息组则较对照组升高。结论心和肺组织HIF1-α表达核阳性细胞的出现,可以作为窒息死亡的特征性表现。     

Examination of the effect of dose-death interval on detection of meperidine exposure in decomposed skeletal tissues using microwave-assisted extraction

The effect of dose-death interval and tissue distribution on the detection of meperidine in selected skeletal tissues was examined using a rapid microwave-assisted extraction (MAE) methodology. Rats (n=14) were dosed with 0 (n=2) or 30 mg/kg (n=12) meperidine (i.p.). Drug-positive rats were sacrificed with CO(2) after 20, 30, 90 and 150 min (n=3 per group). Heart blood was collected immediately after death. Tibiae were excised and frozen for further analysis. The remaining carcasses were allowed to decompose outside in secured cages to the point of complete skeletonization in a rural Northern Ontario location during the late summer months. Vertebrae and pelvi were collected for each animal. Tibial marrow was homogenized in 3 mL PB6 (phosphate buffer, 0.1M, pH 6). Fresh tibiae, and decomposed vertebrae and pelvi were cleaned in PB8.5 (phosphate buffer, 0.1M, pH 8.5) and sonicated to remove remaining soft tissue. Samples of dried, ground bone (0.5-1g) suspended in 2 mL PB6 were then irradiated in a domestic microwave oven (1100 W) at atmospheric pressure for 15 min. Samples of vertebral bone (1g) were also extracted by passive incubation in methanol (3 mL, 50°C, 72 h). All supernatants then underwent solid-phase extraction and analysis by GC/MS, using electron impact ionization in the Selected Ion Monitoring (SIM) mode. Mean GC/MS responses for each tissue type were negatively correlated with dose-death interval, with correlation coefficients ranging from -0.32 to -0.87. Analysis of variance showed dose-death interval to be a main effect (p<0.05) with respect to GC/MS response for blood, marrow, tibial epiphyses prepared by MAE, and vertebral bone prepared by passive extraction, but not for tibial diaphyses, pelvi or vertebrae prepared by MAE. Overall, MAE is advantageous as a rapid extraction tool for screening purposes in skeletal tissues, but assignment of significance to quantitative expressions of skeletal drug concentrations is complex and should be approached with caution.     

Detection of fingermarks by colloidal gold (MMD/SMD)--beyond the pH 3 limit

Dust was investigated for its ability to retain source attribution profiles (SAPs) after chemical exposure. Three distinct sources of the organophosphate pesticide acephate were investigated as a proof-of-concept model. In addition, attribution profiles were created and tested using compounds related to chemical warfare agents (CWAs), specifically VX and G-series agents: O-ethyl methylphosphonothioate (EMPTA), N,N-diisopropylmethylamine (DIPMA), N,N-diisopropylethylamine (DIEA), diisopropylamine (DIPA), diethyl aniline (DEA), diethyl ethyl phosphonate (DEEP), trimethyl phosphite (TMP), dimethyl hydrogen phosphite (DMHP), diethyl hydrogen phosphite (DEHP), triethyl phosphate (TEP), ethyl methylphosphonate (EMPA), and diisopropyl methylphosphonate (DIMP). Dust was collected from a storage shed, aliquots deposited on carpet and loaded with distinct chemical profiles using an exposure chamber and aerosolizer. After a given period of time (1h, 24h, or 72 h), the dust was extracted and its SAP analyzed by gas chromatography-mass spectrometry (GC-MS) and/or liquid chromatography-tandem mass spectrometry (LC-MS/MS). Principal components analysis (PCA) was used to determine the association of dust exposed to the same and different chemical sources. PCA results demonstrate that dust samples exposed to distinct chemical sources are clearly differentiated from one another across all collection times. Furthermore, dust aliquots exposed to the same source can be clearly associated with one another across all collection times. When the CWA-related compounds were subjected to elevated temperature (90°C) conditions, it was found that the signature was stable at the 1h and 24h collections. At 72 h and elevated temperature, larger deviations from the control were observed for some compounds. Elevated pH (10) affected the profile to a lesser degree than elevated temperature. Overall, dust is found to be an effective media for the in situ collection of source attribution profiles.     

大鼠挫伤骨骼肌中M1/M2型巨噬细胞时间相关性研究

目的 研究大鼠骨骼肌挫伤后M1/M2型巨噬细胞浸润的动态变化规律结合组织学改变建立损伤时间推断方法.方法 建立大鼠骨骼肌挫伤模型,挫伤后6h、12h、24h、36h、48h、60h、72h、96h、144h、216h、336h为实验组,对照组不进行任何处理.采用HE染色法和免疫荧光多重染色方法观察骨骼肌挫伤后修复规律,...     

Six‐Year Retrospective Study of Suicidal Hangings: Determination of the Pattern of Limb Lesions Induced by Body Responses to Asphyxia by Hanging*

Abstract: The objective of the present study was to estimate the proportion of hanging victims presenting with limb lesions, to compare this rate between hanging in restraint spaces and in more open settings, and to describe the usual pattern of limb lesions associated with hanging. Two hundred and seven cases of suicidal hanging were retrospectively reviewed and compared to 45 homicidal nonhanging strangulation victims. Bruises incidence was significantly lower in hanging victims (19.8%) compared to homicidal strangulation victims (55.6%). Bruises were more commonly encountered in restraint areas such as closets and staircases (56.3% and 66.7%, respectively) than in more open settings such as barn, bridge, fence, and park. Limb bruises on hanging victims were generally located on the posterior upper limb or the anterior lower limbs, whereas strangulation victims did not display this preferential bruises concentration. Possible suspicion criteria for limb bruises distribution are discussed, in relation to physiopathology of human asphyxia by hanging.     

A preliminary study on the changes of expression of PDGF-beta,PDGFR-beta,TGF-beta 1, TGFR,bFGF and its relationship with the wound age in wound healing

OBJECTIVE: To explore the relationship between the expression change of cytokines and the wound age during the healing process of rats skin wound. METHODS: Immunohistochemical and image-analysis methods were performed on vital skin wounds(after incision 0.5-168 h am) and postmortem damage(after incision 0.5-6 h pm). RESULTS: The expression of the cytokines PDGF-beta, PDGFR-beta, TGF-beta 1, and bFGF in the epithelial cells was already enhanced since 0.5 h am after damage and their strongest expression reaction was seen at 24-96 h am. In addition, the expression of PDGF-beta, PDGFR-beta, TGF-beta 1 and bFGF was also found in the macrophages and the fibroblasts of the granulation tissue, and the expression changes in the postmortem damage group showed that the skin tissue within 0.5-3 h after incision showed immunohistochemical changes but weakly expression and 3 h thereafter no any change was found. CONCLUSION: The expression characteristics of the above mentioned cytokines in wound repair should be related to the wound age and it reminds therefore that they may be used as immunohistochemical criteria for accurate determining the wound age.     

Knee joint injuries as a reconstructive factors in car-to-pedestrian accidents.

Knee joint injuries were found in 214 out of 357 fatal pedestrian victims of traffic accidents (60%). The cross-sections of tibial and femoral epiphyses revealed bone bruises (due to compression and avulsion) and the percentage of victims with knee injuries increased to 80% (in the group of lateral impacts - 94%). The bone bruises in the central tibial and femoral condyles were observed only in victims hit in an upright position. There was a strong correlation between the side of impact on the extremities in medium sized pedestrians (from the front, back, lateral and medial side) caused by passenger cars and the mechanism of knee injuries (hyperextension, anterior dislocation of the proximal tibial epiphysis in relation to the femoral condyles, valgus and varus flexion). In the cases of very low impacts (e.g. in very tall victims hit by rapidly breaking wedge-shaped cars) or very high impact (e.g. in very short victims or truck hits) the "reversed" complexes of injuries were found (lever principle). These findings showed that knee joint injuries are useful for determining the car-pedestrian location on collision and the type of vehicle (in hit-and-run accidents).     

Vital reactions to frostbite of the ear and paw skin in guinea pigs exposed to the cold

Vital reactions to frostbite in the paw and ear skin of guinea pigs were studied in order to find an animal model for frostbite in cases of accidental hypothermia. One group of animals was rendered hypothermic (rectal temperature, 30 degrees C) by exposure to an ambient temperature of -20 degrees C, and samples were taken from the frozen skin. A second group was rendered hypothermic and rewarmed in warm air at 45 degrees C, and samples were taken from the thawed skin. The only vital reaction in the first group (freezing time, 4-5 h) was mild initial inflammation, which was expressed in granulocyte adhesion to the vessel wall and the migration of a few cells into the dermis. The inflammatory reaction was more distinct in the second group (freezing and thawing together 5-7 h), with a large number of granulocytes being present in the dermis. Oedema and hyperaemia were also present in the frostbitten tissue after thawing, but no signs of necrosis developed. The alkaline-phosphatase reaction demonstrated the presence of granulocytes more clearly than H & E or Masson trichrome staining. Vital reactions were more advanced in the ear skin. It is concluded that vital reactions are very scarce in cases of frostbite, even after several hours' exposure, unless the tissue is allowed to thaw.     

What happens in freezing bodies? Experimental study of histological tissue change caused by freezing injuries.

In order to evaluate histological features of freezing damages to human tissue after death, we froze samples of liver and heart tissue to temperatures of -12 degrees C, -28 degrees C and -80 degrees C, and stored them for 24 and 72 h, respectively, at those temperatures. After thawing and routine preparation for histology, the samples were evaluated both by microscope and with an electronic image analyzer. In all cases, we found extended extracellular spaces and shrunken cells resulting from the freeze-thaw cycle. These features were more pronounced in tissues stored for longer durations. Such findings seem to be typical of tissue that has been frozen prior to examination. Two cases of dead bodies found outdoors at subzero temperatures demonstrate that formerly frozen and unfrozen tissues can be distinguished histologically. The findings are examined in relation to the fundamental laws of cryobiology.     

Post-mortem toxico-kinetics of trazodone.

Trazodone is a structurally unique bicyclic anti-depressant, said to be significantly less toxic than other anti-depressants following an acute overdose. We studied the tissue distribution and post-mortem redistribution of trazodone in two fatalities, one of which allowed comparison with trimipramine, a typical tricyclic anti-depressant. Case 1, a 53-year-old female weighing 72 kg, had femoral vein concentrations of trimipramine 5.5 micrograms/ml, trazodone 14.4 micrograms/ml and alcohol 107 mg%. Case 2, a 48-year-old female of 70 kg, had a femoral vein trazodone of 15.5 micrograms/ml and alcohol 34 mg%, with no other drugs detected. For case 1 and case 2 respectively, trazodone tissue concentrations were: skeletal muscle 7.3 and 9.0 micrograms/g; left and right lungs 13.3, 12.9 and 35.3, 40.1; myocardium, 30.9 and 28.9; kidneys 34.7 and 39.6; liver 73.7 and 82.4; fat 18.5 and 16.5; brain 48.6 and 20.9. For case 1 and 2, respectively, blood trazodone concentrations in 10 initial autopsy samples ranged from 13.7-17.3 and 14.4-16.9 micrograms/ml. Twenty-four and forty-eight hours later the respective ranges were 12.8-18.0 and 12.4-19.9 for case 1, 12.5-20.1 and 12.7-27.0 for case 2. By contrast, for trimipramine, blood concentrations at 0 time, 24 h and 48 hours ranged from 5.5-11.4, 5.2-14.3, and 4.2-18.2, respectively. We conclude that trazodone shows little preferential concentration in solid organs and consequently has relatively stable post-mortem blood concentrations with little drug redistribution artefact. Both the clinical pharmacokinetics and post-mortem toxicokinetics of trazodone differ significantly from the tricyclic anti-depressants.