C. Hernández-Cueto

Advances in the diagnosis of wound vitality: a review.

The diagnosis of the vital origin of wounds in many cases remains an unsolved problem for the forensic pathologist. Practical experience enables the expert to diagnose the vital or postmortem origin of wounds on the basis of macroscopic examination. In some cases, optic microscopy is used to confirm the diagnosis. In many other cases, additional more sensitive and specific markers of vitality are required. In the past 50 years, comprehensive research on this topic has resulted in a better understanding of the acute inflammatory reaction. The development and application of sensitive and specific markers through research in the areas of histochemistry, enzymology, and biochemistry has provided a partial solution to the problems involved in wound vitality diagnosis. A review of this challenging area of forensic pathology, including an explanation of these methods and markers, is presented in this paper.

The usefulness of lung surfactant phospholipids (LSPs) in the diagnosis of drowning

The authors have studied the usefulness of some lung surfactant phospholipids (LSPs) isolated from lung tissues as markers of drowning. Two different groups of rabbits were sacrificed by drowning in fresh and salt water, and their phospholipid compositions were compared with those of a non-drowned control series. For the phospholipids studied in lung lavages (phosphatidyl choline, phosphatidyl ethanolamine, and phosphatidyl glycerol) the proportions differed between the control group and the drowned group, and between the fresh-water and salt-water drowned animals. According to these results, the lipids we have analyzed can be employed as markers in forensic autopsies, where it is necessary to differentiate between death by drowning and postmortem immersion and between fresh-water and salt-water drowning. In lung tissue, only phosphatidyl choline and phosphatidyl inositol showed significative differences. These results also confirm that LSPs are strongly affected in drowning.

Plasmatic levels of atrial natriuretic peptide (ANP) in drowning. A pilot study

Bodies found in water may cause problems for forensic pathologists who have to differentiate drowning from postmortem immersion or fresh from salt water drowning. The exact physiopathology of drowning is still controversial and complementary tests can not exactly establish the exact cause of death if macroscopic findings at autopsy are not conclusive. We have employed atrial natriuretic peptide (ANP) as a marker in an experimental series of fresh and salt water drowning, comparing their results with a non-drowned control series. There are differences between the plasma basal levels of the control series (79 pg/ml) and the levels in animals drowned in fresh water (358 pg/ml, P less than 0.001) and between control and rabbits drowned in salt water (190 pg/ml, P less than 0.001). According to these values, there are also differences between fresh and salt water drowned animals (P less than 0.001). We propose this peptide as a new marker in cases of drowning, with the ability to differentiate drowning from postmortem immersion and fresh from salt water drowning.

Padova Charter on personal injury and damage under civil-tort law Medico-legal guidelines on methods of ascertainment and criteria of evaluation

Compensation for personal damage, defined as any pecuniary or non-pecuniary loss causally related to a personal injury under civil-tort law, is strictly based on the local jurisdiction and therefore varies significantly across the world. This manuscript presents the first “International Guidelines on Medico-Legal Methods of Ascertainment and Criteria of Evaluation of Personal Injury and Damage under Civil-Tort Law”. This consensus document, which includes a step-by-step illustrated explanation of flow charts articulated in eight sequential steps and a comprehensive description of the ascertainment methodology and the criteria of evaluation, has been developed by an International Working Group composed of juridical and medico-legal experts and adopted as Guidelines by the International Academy of Legal Medicine (IALM).

Free amino acid concentrations in vitreous humor and cerebrospinal fluid in relation to the cause of death and postmortem interval.

Abstract:  We studied free amino acids in vitreous humor and cerebrospinal fluid from 58 cadavers in the course of routine medicolegal autopsies in the city of Granada. The main objective was to establish whether free amino acids contents in these fluids were related with the cause of death, postmortem interval, and severity of the classic signs of asphyxia. The amino acids (aspartic acid, glutamic acid, serine, glutamine, glycine/threonine/histidine, citruline, arginine, alanine, taurine, GABA, tirosine, valine, methionine, isoleucine, phenylalanine/tryptophan, leucine, and lysine) were quantified by high performance liquid chromatography. There were no statistically significant differences in amino acids concentrations in vitreous humor when the different causes of death were considered. Our results did not show any statistically significant relationship when asphyxial score was plotted against the vitreous content of each amino acid. A statistically significant increase with postmortem interval was observed in vitreous taurine (r = 0.3191, p = 0.01461), glutamate (r = 0.4323, p = 0.0007) and particularly in aspartate (r = 0.4508, p = 0.0003).

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 injury. 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 as a vitality marker in human skin wounds

SummaryThis paper shows the results obtained by studying the lysosomal enzyme Cathepsin D as a potential marker for the vitality of wounds in human specimens. We have analyzed 53 samples using enzymological and histological techniques. Our results show the ability of Cathepsin D to establish the vital origin of wounds inflicted 5 minutes or less before death, where the specific activity of cathepsin D reached 0.055 units at the wound edge and 0.01 units in their respective controls (P < 0.001). As previously demonstrated in an experimental series, Cathepsin D seems to be a very useful marker of high forensic interest in especially difficult cases. Further studies are in progress to check the influence of different factors such as drugs intake and clinical conditions on Cathepsin D activity.ZusammefassungDas Manuskript zeigt Ergebnisse, wie sie durch Untersuchung des lysosomalen Enzyms Cathepsin D als potentiellen Marker für Vitalität in menschlichen Hautwunden erzielt wurden. Wir haben 53 Proben mit Hilfe enzymatischer und histologischer Techniken untersucht. Die Ergebnisse zeigen die Fähigkeit von Cathepsin D, den vitalen Ursprung von Wunden zu etablieren, selbst wenn sie 5 Minuten oder geringere Zeit vor dem Tod gesetzt wurden. In den letzteren erreichte die spezifische Aktivität von Cathepsin D 0,055 Einheiten an den Wundränden und 0,01 Einheiten in den entsprechenden Kontrollen (P < 0,001). Wie früher in experimentellen Untersuchungen gezeigt wurde, scheint Cathepsin D ein sehr nützlicher Marker von hoher forensischer Bedeutung besonders in schwierigen Fällen zu sein. Weitere Studien sind in der Bearbeitung, um den Einfluß verschiedener Faktoren, wie Medikamenteneinnahme und klinischer Zustand, auf die Cathepsin D-Aktivität zu überprüfen.

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.

Prostaglandin F2a (PGF2a): an inadequate marker of the vitality of wounds?

SummaryWe have studied the viability of (PGF2a): as a vitality marker in skin wounds. Incised vital skin wounds and homolateral control pieces of skin were obtained from 20 autopsies performed at the Institute of Legal Medicine of Coimbra University (Portugal). We have also studied 10 fresh skin samples from the Department of Dermatology of the University Hospital (Granada). Our results show that (PGF2a): is not suitable for the diagnosis of the vitality of wounds because of its irregular behaviour.ZusammenfassungDie Verwendbarkeit von Prostaglandin F2a als Vitalitätsmarker wurde an Hautverletzungen studiert. Als Material dienten 20 vitale Schnittverletzungen sowie entsprechende Kontrollen unverletzter Haut aus 20 Obduktionsfällen des Instituts für Rechtsmedizin der Universität Coimbra (Portugal). Weiter wurden 10 frische Hautproben von Patienten der Dermatologischen Abteilung des Klinikums Granada untersucht. Die Prostaglandin-Analysen wurden mittels RIA durchgeführt. Die Ergebnisse belegen, daß die Bestimmung des (PGF2a): nicht als Methode des Vitalitätsnachweises von Wunden geeignet ist.

Diagnostic ability of D-dimer in the establishment of the vitality of wounds

The potential of skin measurements of D-dimer (DD), a specific derivative of crosslinked fibrin, for the diagnosis of the vital origin of wounds was investigated in 67 human vital skin wounds and homolateral control pieces of skin from 53 autopsies performed at the Institute of Legal Medicine of Coimbra (Portugal) during 1992. Our results demonstrate that DD levels for the incised vital skin wounds (n = 50) were much higher on average than their controls (P < 0.01) which were actually incised injuries produced postmortem at the autopsy. We did not find any statistical difference in average DD levels among the remaining injuries (seven abrasions and ten contusions) in comparison to their respective controls. DD may be used, with certain limitations, as a marker of the vitality of incised wounds. The authors believe this is the first time this marker has been applied in forensic pathology. It is an example of an easy and quick application technique, with the added advantage of low cost.