Tool mark striations in pig skin produced by stabs from a serrated blade

Stab wounds produced by serrated blades are generally indistinguishable from stab wounds produced by non-serrated blades, except when visible tool mark striations are left on severed cartilage. Using a pig-skin experimental model, we explored the possibility that similar striations may be left in skin. Stabs into pig skin were made using a straight spine coarsely serrated blade (121), a drop point finely serrated blade (20), a clip point irregular coarsely serrated blade (20), a drop point coarsely serrated blade (15), and as controls 2 non-serrated blades (40). Tool mark striations could be seen on the skin wall of the stab canal in all stabs made using serrated blades but in none with non-serrated blades.The striation pattern, reflecting the class characteristics of the serrated blade, was the same as that described in cartilage but less well defined. Fixation of the specimen with Carnoy's solution best preserved visible striations, and fixation with formaldehyde after staining with 5% Neutral Red was also satisfactory. Casting with vinyl polysiloxane dental impression material greatly facilitated photo-documentation. Applying the technique to homicidal stabbings may help identify stab wounds produced with serrated blades.     

Expression of adhesion molecules in skin wounds: diagnostic value in legal medicine

The adhesion molecules identified in recent years can help improve the diagnosis of the wound age, especially of injuries with a short survival time. This is also indicative of the vitality of the wounds. The material investigated in the study originated from 465 skin wounds. The samples were taken from human autopsy material, during the surgical treatment of wounds (excision) of patients and from experimental incised wounds of mice. To judge the age of skin wounds the endothelial adhesion molecules were detected in paraffin sections after autoclaving and using the ABC technique. Human skin wounds: strong positive staining was observed of ICAM-1 1.5 h at the earliest and 3.5 days at the latest, for the P-selectin 3 min at the earliest and 7 h at the latest, for the E-selectin 1 h at the earliest and 17 days at the latest and for VCAM-1 3 h at the earliest and 3.5 days at the latest after the time of injury. The L-selectin was expressed constitutively. Mice skin wounds: strong positive immunohistochemical reactions were found as a rule earlier than in human skin wounds. The detection of an increased expression of ICAM-1, VCAM-1 and P- and E-selectins can improve the wound age assessment in injuries with short survival times.     

Release of proteinase inhibitors as a vital reaction in the early post-traumatic interval

In a pilot study paraffin-embedded sections of open skin wounds (stab and slash wounds, lacerations) were investigated to determine the presence of a vital reaction. Granulocytes were detected by naphthol AS-D chloroacetate esterase, the enzyme "lysozyme", and eight proteinase inhibitors by the indirect immunoperoxidase method. The tissue specimens were taken from consecutive autopsy material. The survival time could be determined in 14 cases (10-165 min) and was unknown in 12 other cases of sudden death due to injury of the major vessels or heart. The controls were cases with injuries inflicted after and cases of sudden death due to massive blunt trauma served death. In vital injuries, accumulations of proteinase inhibitors, particularly alpha-2-macroglobulin and alpha-1-antichymotrypsin, were demonstrable in the corium parallel to the wound surface. In comparison, the reaction of proteinase inhibitors that neutralize only enzymes participating in blood coagulation or complement activation (C1-esterase inhibitor and protein C) was absent or weak. Protein accumulation was observed only sporadically in cases of sudden death and never in cases with wounds inflicted after death. No relationship could be established between semiquantitatively estimated staining and survival time. Granulocytes and lysozyme were first observed in the corium after a survival time of more than 60 min.     

大鼠皮肤切创愈合过程中泛素的表达及其法医学意义

目的观察大鼠皮肤切创后泛素(ubiquitin)表达的变化规律。方法应用免疫组化方法,观察大鼠皮肤切创后1、3、6、12h和1、3、6、10、14d ubiquitin的表达情况,并用图像分析系统进行图像分析。结果正常对照组大鼠皮肤有低水平ubiquitin表达,切创后ubiquitin表达增加,伤后6d达峰值,伤后10d开始下降,伤后14d恢复到正常水平。结论ubiquitin可作为法医学损伤时间推断的有效指标。     

Immunohistochemical detection of fibronectin and tenascin in incised human skin injuries

Immunohistochemical detection of molecules involved in inflammatory reaction can be useful for the diagnosis of vitality in skin wounds. We studied the expression of fibronectin (FN) and tenascin (TN) in 58 human skin wounds (48 vital and 10 postmortem). The age of vital injuries ranged from 3 min to 8 h and postmortem specimens were collected after a postinfliction interval of 15-180 min. One hundred thirty-seven formalin-fixed paraffin-embedded sections (mean: 2.3 sections per case) were stained with each of two monoclonal antibodies against FN and TN using the streptABC technique. A reticular staining for FN in wound edge and dermis was observed in 50% of vital specimens versus 0% in postmortem cases. Immunoreactivity was reduced in 10 autolysed cases. FN positivity exclusively at the injury margin was observed in 39.4% of vital wounds and 10% of postmortem cases. TN was negative in all specimens. Vital and postmortem hemorrhage areas showed positivity for FN and TN. Due to its low sensitivity, immunohistochemical analysis of FN is useful for determining vitality only in a minority of cases. Different factors in everyday practice, including autolysis and technical problems often produce false negative reactions with the result that FN cannot be regarded as a reliable parameter of vitality. Positive reactions (network staining) are more valuable than negativity but are not pathognomonic. Both vital and postmortem hemorrhages show an enhanced positivity for FN and TN, thus impeding the diagnosis.     

细胞因子(IL-6、TNF-α)在损伤时间中原位杂交法研究

采用原位杂交法，研究大鼠切削皮肤中细胞团子IL－6、TNF－αmRNA表达量，旨在探讨IL－6、TNF－α推断法医损伤时间的应用价值以及其在损伤生活反应中的分子机制。研究结果表明，根据大鼠损伤皮肤中TNF－αmRNA表达量能够区别生前伤与死后伤，并可以利用TNF－αmRNA表达量改变准确区别60min内和60min后损伤时间，但IL－6mRNA表达量在研究组内均未见阳性反应，不能推断损伤时间。     

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 degrees 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.     

应用PAP法检测成人皮肤切创纤维连接蛋白推断损伤时间

应用PAP法，对15例成人腹部不同时间组（2小时以内）手术切创皮肤进行研究，观察Fn的分布、含量变化及其与损伤时间的关系；同时作H．E和Masson三色染色对比观察。结果发现：创伤即刻，创壁仅有少量Fn阳性颗粒；15分钟后，创壁表面Fn含量增多，呈薄层、断续的细条状；以后，随着时间延长，创壁表面Fn含量逐渐增多，呈连续细带状；至2小时，创伤局部Fn含量明显增多，创壁表面Fn是较宽带状，创壁内Fn亦增多，并是浓度梯度改变。PAP法检测Fn，可为鉴别生前与死后损伤、推断早期损伤时间，提供新的手段。     

Peroxidase activity in traumatic skin lesions

Peroxidase activity was determined in experimental compression-excoriation lesions and incision wounds of rat skin after different periods of vital time. The peroxidase enzyme was extracted from the tissues by homogenization in 0.5% cetyltrimethylammoniumbromide, and the enzyme activity was measured from the supernatant by o-dianisidine-H2O2 assay. In the blood of the rats a mean activity of approx. 5.26 +/- 1.11 U/g dry weight was observed. In the control specimens of the skin the activity was very low and generally below the detection limit of the methods used. In 30-min-old compression-excoriation lesions the mean peroxidase activity was 0.38 +/- 0.21 U/g dry weight. In lesions older than 30 min the activity started to increase rapidly. In 4-h-old compression-excoriation lesions it was 10 times higher than the 30-min level and was 40 times higher in 12-h-old lesions and 70-100 times higher in 1-3-day-old compression-excoriation lesions, respectively. In 30-min-old incision wounds the mean peroxidase activity was 0.65 +/- 0.37 U/g dry weight. The increase of the activity compared with the 30-min level was even faster in the incision wounds: in 4-h-old wounds the mean activity was 50 times higher, in 12-h-old wounds 200 times higher and in those of 1-5 days it was several hundreds of times higher. Compression-excoriation lesions made after death showed activity similar to the control specimens. Postmortem autolysis at +22 degrees C resulted in a loss of the enzyme activity in 1-day-old compression-excoriation lesions so that after 3 days approx. 80% remained, and after 5 and 7 days approx. 40% was present. After 3 days of autolysis at +4 degrees C, nearly 100% of the activity remained and approx. 90% was present after 5 and 7 days of autolysis. Increased peroxidase activity was also detectable in human vital excoriations in the specimens which were taken in autopsies several days postmortem.     

Morphologic, experimental-comparative investigation as an identification of the injuring instrument method

Aiming to identify the injuring tool characteristics and the tool itself morphologic, experimental-comparative investigations of the skin wound, rib and cartilage injuries taken during the autopsy are performed. During 1995-2004, 489 investigations were performed for this purpose. In 418 cases, knives were submitted for identification of the specific injuring tool (in total-835 knives). In 205 cases the investigation included not only skin wounds, but also the injured rib cartilages. Identification investigations were performed by investigating both the skin wounds morphologic characteristics and dynamic traces-trails in the rib cartilage tissue left by the micro relief of the knife blade edge. In the case of the investigated and experimental skin wounds characteristics coincidence the experimental and comparative dynamic traces investigation was performed when the traces were suitable for the tool identification purposes. In the case of the investigated and experimental skin wounds, dynamic traces coincidence, the totality of the coincided characteristics was considered individual. In those cases, the conclusion included the fact that the injury had been made by a particular knife. According to our data during 1995-2004 in 23 cases-15.9% (5.5% out of the total investigated cases), the knife identification was based on the skin wound characteristics and dynamic traces in the rib cartilage tissue. In our opinion, the dynamic traces in the rib cartilage tissue investigations supplement the identification field and are valuable in the tool identification. In 11 (2.6% out of the total investigated cases) cases knives were identified only by the skin wounds morphologic characteristics, the ribs being not injured or dynamic traces being not suitable for the tool identification.     

The significance of collagen in determining the age of a wound

The aim of the present study was to find out whether the quantitative and/or qualitative analysis of collagen can be useful in the determination of the age of healing skin wounds in various body regions of patients of different ages and sex. The quantitative measurement of total wound collagen revealed no clear time-dependent changes. Immunohistochemical staining for collagen types I and III demonstrated the presence of type III as early as 2 days after injury. Procollagen type I was found in wounds beginning from 4 days on, while type I collagen was not present before 6 days after the injury. Immunohistochemical analysis of specific basement membrane proteins, collagen type IV and laminin, showed a reconstruction of the epithelial basement membrane beginning from on day 5, while a completely rebuilt basement membrane was found in most cases after more than 14 days, depending on the dimension of the wound and its treatment.     

IL-33在小鼠皮肤创伤后损伤时间推断中的作用

目的通过观察白细胞介素-33(interleukin-33,IL-33)在皮肤创伤后的时序性变化规律,探讨IL-33在法医学实践中用于损伤时间推断的应用价值。方法利用直径为5mm的圆形锉刀在小鼠背部建造皮肤损伤模型,于伤后1h、3h、6h、12h、1d、3d、5d、7d、10d取损伤处组织,对照组在与创伤组小鼠相同部位取同等大小的皮肤样本。采用苏木精-伊红(hematoxylin-eosin,HE)染色法观察皮肤创伤后愈合过程中的形态学变化,通过Western印迹法、免疫组织化学染色和双重免疫荧光染色法检测皮肤创伤样本IL-33的表达变化。结果Western印迹法结果显示,伤后3h,IL-33蛋白表达稍有下降,6h后IL-33蛋白表达逐渐增加,于伤后3d达峰值,随后逐渐减少。免疫组织化学染色结果显示,在对照组皮肤的表皮、毛囊、皮脂腺及真皮中固有细胞有少量IL-33阳性表达,伤后3h,IL-33阳性细胞率开始增加,伤后3d达到峰值,随后逐渐减少。双重免疫荧光染色结果显示,伤后1~3d,IL-33阳性表达细胞主要为巨噬细胞,伤后5~7d,IL-33阳性表达细胞主要为肌成纤维细胞。HE染色结果显示该皮肤损伤模型创口愈合过程符合炎症的病理学发展规律。结论IL-33有望成为法医学推断皮肤损伤时间的参考指标。     

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 stage, 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 (less than 0.01). Multiple range test shows that significant differences (P less than 0.05 or P less than 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.     

Finger prints on human skin--results of a general applications-oriented series

Numerous methods have been described for making dactyloscopic traces visible on human skin, but these have never become routine--last but not least because they are too complicated and time-consuming to perform. The purpose of the present investigation was to find out to what extent fingerprints on the skin of deceased persons can be made visible with the help of conventional methods like iodine fuming or brushing with Magna-Brush and can be preserved with gelatine film or moulding material. Of the total number of 486 experimental fingerprints the dermal ridges could be made visible in 150 prints. The best findings were obtained when the trace had been applied several hours post mortem. On the skin of the extremities better results could be achieved than on the trunk or the neck. Dactyloscopic tracing on the skin of bodies in homicide cases should be taken into consideration especially if there is evidence that the perpetrator touched the victim after his/her death (e.g. if the body was transported).     

The time-dependent expression of fibronectin,MRP8, MRP14 and defensin in surgically treated human skin wounds

Fibronectin, MRP8, MRP14 and defensin were detected immunohistochemically in 46 surgically treated, fresh (hours old) human wounds and in wounds of 13 individuals who died immediately from fatal trauma (airplane crashes or train rollovers). In immediate fatal trauma, it was not possible to detect fibronectin outside of bleeding areas nor could inflammatory cells be visualized in the interstitium using MRP8, MRP14, or defensin antibodies. Fibronectin staining could be regularly demonstrated in wounds at least 20 min-old. Granulocytic infiltrates limited to the perivasal space could be detected 20-30 min after infliction of the wound expressing MRP8, MRP14 and defensin. It was also possible to detect fibronectin networks and MRP8-, MRP14-, and defensin-positive granulocytes and macrophages in particular wounds up to 30 days-old. No differences between the expression of MRP8 and MRP14 could be demonstrated in the wounds, the majority of which were only several hours old. As wound age increased, the number of defensin-positive granulocytes detected decreased. The immunohistochemical detection of fibronectin is a useful way to demonstrate vitality in fresh wounds, beginning about 20-30 min post-trauma. However, detection of MRP8, MRP14 and defensin provides no advantage over the routine histological detection of granulocytes and macrophages in wounds under 1-2 days old.     

Quantitative analysis of proinflammatory cytokines (IL-1beta, IL-6, TNF-alpha) in human skin wounds

Proinflammatory cytokines play an important role in the mediation of inflammation and trauma. They could be useful for the determination of vitality and wound age. In the present study, 144 human skin wounds due to sharp force were investigated. The material was collected during operations (N=96) and postmortem examinations (N=48). The wound age varied from several seconds or minutes to 9 days. Control skin was available in each individual. The tissue specimens were homogenized and extracted in a solution of PBS and protease inhibitors. Interleukin-1beta (IL-1beta), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-alpha) were measured by quantitative ELISA analysis. Statistical evaluation was performed by the t-test using the quotients of levels (wound sample/control skin). In surgical specimens the cytokine levels revealed a clear tendency to increase with wound age. IL-1beta in early skin wounds (24 h, P<0.05). The quantitative analysis of proinflammatory cytokines in wound extracts can contribute to the determination of vitality and wound age, in particular in the very early post-traumatic interval (classic stab wounds).     

损伤经过时间的实验研究

作者在全身麻醉及无菌条件下,通过在大白鼠背部皮肤造成的切创创口,于伤后不同时间杀死.每组中1只于处死前8小时注射秋水仙碱以观察核分裂.取损伤处皮肤,经福尔马林固定、石蜡切片、HE染色、Gorden和Sweet染色(网状纤维)、Weigert染色(弹力纤维)、PTAH染色(纤维蛋白)和Van Gieson染色,于显微镜下观察.结果发现创口逐日缩小,但其中24小时创口较12小时创口有所扩大,伤后第6～7日创口呈突击式缩小.通过组织学变化观察,作者对创口收缩现象在判断损伤时间的意义及形成机理作了讨论.     

对纤维蛋白诊断生前伤的评价——45例枪弹创组织的MSB染色和扫描电镜对比观察

生活反应是诊断生前伤的主要根据.近二十年来,法医病理学在研究生前伤和死后伤的诊断中,采用了多种新兴技术,如扫描电镜技术、酶组织化学技术、酶标技术、电泳技术等,促使生前伤的诊断水平有明显提高.伤后存活1小时死亡者,大部分皆可获得明确诊断;立即死亡或在濒死期形成的损伤,由于生活反应甚微,或受腐败等     

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 degrees 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.     

Intra- and perioral shooting fatalities.

Determination of the manner of death in the case of intra- and perioral firearm wounds can be difficult especially if death scene investigation is unclear and inadequate. In this study, we investigated some characteristics of these firearm wounds which were autopsied in Istanbul. During the 5-year period from 1991 through 1995, there were 15 intra- and perioral firearm fatalities investigated. In all the cases, only one shot was fired into the mouth. They constituted 1% of all the firearm fatalities. The mean age of the victims was 27 years and males constituted 73.3% of the victims. Most of the wounds were caused by handguns. Homicides accounted for 53.3% of these deaths. Three of 15 cases could not be identified as intraoral firearm wounds by general practitioners during the scene investigations.