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  • 1
    ISSN: 1437-1596
    Keywords: Key words Forensic pathology ; Wound age ; determination ; Immunohistochemistry ; Morphometrical analysis ; Interleukin-1α (IL-1α)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Notes: Abstract An immunohistochemical and morphometrical study on the temporal expression of interleukin-1α (IL-1α) was performed on 40 human skin wounds with different wound ages. Immunohistochemically, polymorphonuclear neutrophils mainly showed positive reactions for IL-1α in wounds aged between 4 h and 1 day, but with increasing wound age, neutrophils were no longer present, and macrophages and fibroblasts were positively stained. Morphometrically, the ratio of the number of IL-1α-positive infiltrating cells to the total number of infiltrating cells was evaluated. A considerable increase in the IL-1α-positive cell ratio was observed in wound specimens aged 4 h to 1 day, and the maximum ratio was 46.5% in a 7 h-old wound. The mean value of the IL-1α-positive ratio in 10 wound specimens with different wound ages between 4 h and 1 day was 32.8 ± 9.7%. In most cases the ratio of IL-1α-positive cells gradually decreased in wounds aged between 1.5 and 21 days to less than 30%, and the mean value was 17.5 ± 7.2% (n = 27). These results suggest that ratios of IL-1α-positive cells considerably exceeding 30%, indicate a postinfliction interval of 1 day or less.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 105 (1993), S. 229-232 
    ISSN: 1437-1596
    Keywords: Reepithelialization ; Keratin 5 ; Keratin 13 ; Wound age ; Immunohistochemistry ; Reepithelialisierung ; Keratin 5 ; Keratin 13 ; Wundalter ; Immunhistochemie
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung An 55 chirurgisch versorgten menschlichen Hautwunden mit einer Überlebenszeit zwischen 8 Stunden und über 2 Monaten wurde die wundaltersabhängige Expression von Keratin 5 und 13 während der Reepithelialisierung untersucht. Eine vollständige Dekkung des ursprünglichen Epitheldefektes war erstmals in einer 5 Tage alten Hautwunde zu beobachten, wobei in der Basalzellschicht nur einzelne Cytokeratin 5-positive Zellen nachweisbar waren. Nach 13 Tagen Überlebenszeit war hier erstmals eine vollständige Anfärbbarkeit für Cytokeratin 5 feststellbar, regelhaft war dies nach mehr als ca. 23 Tagen Überlebenszeit der Fall. Nach einem Wundalter von ca. 18 Tagen war die Reepithelialisierung des ursprünglichen Defektes immer vervollständigt, auch wenn nicht alle Basalzellen Cytokeratin 5 exprimierten. Die Darstellung von Cytokeratin 13, ein für nicht-verhornendes Plattenepithel typischer Marker, liefert keine Informationen, die für eine Altersbestimmung menschlicher Hautwunden verwertbar wären, da offensichtlich eine relevante temporäre Expression während der Wundheilung in der menschlichen Haut nicht erfolgt.
    Notes: Summary The time-dependent reepithelialization of 55 human surgical skin wounds with a wound age between 8h and more than 2 months was investigated by the immunohistochemical localization of cytokeratins 5 and 13. A complete, rebuilt epidermal layer over the wound area was first detectable in a 5-day-old wound, while all wounds of more than 18 days duration contained a completely reepithelialized wound area. Between 5 and 18 days the basal layer of keratinocytes showed — in contrast to normal skin — only some cells positive for cytokeratin 5. In some, but not all lesions with a wound age of 13 days or more, a basal cell layer completely staining for cytokeratin 5 was demonstrable. This staining pattern was found in all skin wounds with a wound age of more than 23 days. The immunohistochemical detection of cytokeratin 13 which can be observed regularly in non-cornifying squamous epithelia provides no information for the time-estimation of human skin wounds, since no significant temporary expression of this polypeptide seems to occur during the healing of human skin wounds.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1437-1596
    Keywords: Collagen type III ; Collagen type V ; Wound age ; Immunohistochemistry ; Kollagen III ; Kollagen V ; Wundalter ; Immunhistochemie
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Es wurden insgesamt 79 chirurgisch versorgte menschliche Hautwunden untersucht und Kollagen Typ III und V immunhistochemisch dargestellt. Netzwerk-artige und Kollagen Typ III-positive, mit Fibroblasten des Wundgebietes assoziierte Strukturen waren erstmals nach 2,5 Tagen Überlebenszeit nachweisbar und traten ab einem Wundalter von mehr als 5 Tagen regelmäßig auf. Eine entsprechende Reaktion für Kollagen Typ V war erstmals nach 3 Tagen und damit etwas später als für Kollagen III zu beobachten. Regelmäßig war eine entsprechende Reaktion ab einem Wundalter von 6 Tagen feststellbar. Der positive immunhistochemische Nachweis von Kollagen Typ III bzw. Typ V belegt somit ein Wundalter von mindestens 2–3 bzw. 3 Tagen, das Fehlen positiver Reaktionen weist auf eine Überlebenszeit von weniger als ca. 6 Tagen hin. Beide Kollagen-Typen waren auch in älteren Wunden (Wundalter 2,5 Monate) nachweisbar, eine weitere Differenzierung des Alters von länger überlebten Verletzungen ist jedoch wegen der Abhängigkeit der längsten Nachweisbarkeit einer reaktiven Veränderung und der ursprünglichen Ausdehnung der Wundfläche nicht möglich.
    Notes: Summary Collagen type III and V were visualized immunohistochemically in 79 surgically treated human skin wounds with a wound age between 8 h and 2.5 months. Network-like structures positively staining for collagen type III and associated with fibroblastic cells in the wound area were first detectable in a 2.5-day-old skin lesion and occurred regularly in wounds more than 5 days old. Collagen type V appeared first in the wound area after about 3 days, slightly later than collagen type III, and was detectable regularly in wounds with a survival time of 6 days or more. The immunohistochemical detection of collagen type III or type V thus indicates a wound age of at least 2–3 days. The lack of a positive reaction in a sufficient number of specimens indicates a wound age of less than 6 days. Even though both collagen types could also be detected in older wounds (wound age 2.5 months), further information for the time-estimation of older skin wounds cannot be given due to the observation that the time period during which reparative processes can be observed depends on the extent of the wound area.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 107 (1995), S. 197-200 
    ISSN: 1437-1596
    Keywords: Macrophages ; Wound age ; Immunohistochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Insgesamt wurden 117 vitale Hautwunden (Überlebenszeit wenige Sekunden bis 7 Monate), 20 postmortal gesetzte Verletzungen sowie Haut mit leichten bzw. fortgeschrittenen Fäulnisveränderungen untersucht und verschiedene Marker der Makrophagen-Differenzierung (27 E 10, RM 3/l, 25 F 9 und G 16/1) analysiert. Der „early stage inflammation marker” 27 E 10 färbte neben Makrophagen auch Monozyten und neutrophile Granulozyten, die innerhalb von Blutgefäßen bzw. in postmortal gesetzten Blutungen lokalisiert waren und liefert somit keine Informationen zum Wundalter, die über die Möglichkeiten des Routine-histologischen Nachweises von Makrophagen hinausgingen. Die von den Antikörpern RM 3/1 (intermediate stage inflammation marker) und 25 F 9 (late stage inflammation marker) erkannten Antigene wurden ausschließlich von Histiozyten und reaktiv eingewanderten Makrophagen exprimiert. Die morphometrische Analyse ergab positive Ergebnisse (definiert als ein mindestens zweifacher Anstieg der Zellzahl in zwei oder mehr Gesichtsfeldern verglichen mit der maximal feststellbaren Zahl an Histiozyten in unverletzter Haut) bei Verwendung der Antikörper RM 3/1 bzw. 25 F 9 frühestens 7 bzw. 11 Tage nach Wundsetzung. Ab 12 Tagen Wundalter reagierte der „chronic stage inflammation marker” G 16/1 erstmals positiv. Das Antigen ließ sich insgesamt allerdings in einem geringeren Prozentsatz der untersuchten Wunden darstellen. Vorteilhaft ist jedoch das Fehlen einer relevanten Expression durch Histiozyten, wodurch die Auswertung der Präparate erleichtert wird. Die entsprechenden Antigene lassen sich zudem in leicht - wenn auch in einer deutlich geringeren Färbeintensität -, aber nicht forgeschritten fäulnisveränderter Haut nachweisen, so daß deren immunhistochemische Darstellung gegebensfalls auch zur Beurteilung von länger überlebten Verletzungen an Leichen mit etwas fortgeschrittener Liegezeit herangezogen werden kann.
    Notes: Abstract 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. The markers 27 E 10, RM 3/1 and 25 F 9 are also useful for the evaluation of slightly - even though the staining intensity was considerably reduced - but not advanced putrefied skin. Therefore, the immunohistochemical analysis of the corresponding antigens can possibly contribute to an age estimation of wounds with advanced post infliction intervals obtained from corpses with longer - but limited - postmortem intervals.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 108 (1996), S. 262-264 
    ISSN: 1437-1596
    Keywords: Wound age ; Morphometry ; Hemosiderin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Notes: Abstract A morphometrical analysis of the extent of hemosiderin deposits in 71 human skin wounds with post-infliction intervals between 2 days and 7 months was performed. Earliest positive findings were detectable in a lesion aged 3 days, and with increasing wound age an increase in the amount of hemosiderin occurred. A value of more than 20% of the microscopic field with hemosiderin deposits was found earliest 8 days after wounding and therefore the detection of considerable amounts of hemosiderin (arbitrarily defined as 20% or more of the evaluated area) indicates a minimum wound age of approximately 1 week. Since the extent of hemosiderin formation depends upon the extent of the initial hemorrhage and a “physiological” reduction in the amount of this pigment with advanced wound age, slight or absent hemosiderin deposits cannot provide information on the post-infliction interval.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1437-1596
    Keywords: Fibronectin ; Immunohistochemistry ; Wound age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Es wurden 53 vitale Hautwunden mit einem Wundalter von wenigen Sekunden/Minuten
    Notes: Summary 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 dit 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 upt 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. Since a faint positive fibronectin staining is seen in postmortem lesions and bleedings, we propose that only those wounds which show strong positive fibronectin staining also extending into the adjacent stroma should be regarded as vital.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    International journal of legal medicine 105 (1992), S. 99-103 
    ISSN: 1437-1596
    Keywords: Myofibroblasts ; Alpha-smooth muscle actin ; Desmin ; Immunohistochemistry ; Wound age ; Myofibroblasten ; Alpha-Aktin ; Desmin ; Immunhistochemie ; Wundalter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Es wurden 66 menschliche Hautwunden mit einem Wundalter zwischen 20 Stunden und 7 Monaten sowie komplikationsloser Wundheilung ausgewertet. Nach immunhistochemischer Darstellung von alpha-Aktin und Desmin wurde das zeitabhängige Auftreten positiv reagierender Myofibroblasten im Wundgebiet untersucht. Es zeigte sich hierbei, daß in Hautwunden mit einem Wundalter unter 5 Tagen keine positiv anfärbbaren Zellen zu beobachten waren. In 57% (25 von 44 Fällen) der Hautverletzungen, die zwischen 5 und 31 Tagen überlebt worden waren, fanden sich im Granulationsgewebe alpha-Aktin haltige Myofibroblasten. Besonders zahlreiche, positiv reagierende Zellen traten zwischen ca. 16 bis 31 Tagen nach Wundsetzung auf, konnten jedoch auch bereits in Hautwunden jüngeren Alters beobachtet werden. In 2 von 7 Fällen mit einem Wundalter zwischen 1 und 7 Monaten (29%) liesen sich ebenfalls alpha-Aktin positive Myofibroblasten im Wundgebiet nachweisen. Desmin-haltige Myofibroblasten konnten nicht beobachtet werden. Die Ergebnisse zeigen, daß alpha-Aktin positive Myofibroblasten bereits mit Ausbildung typischen Granulationsgewebes ab ca. dem 5. Tag nach Verletzung im Wundgebiet auftreten. Der Nachweis positiv reagierender Zellen im Wundgebiet läßt jedoch aufgrund der Variabilität der Befunde keine weitere Differenzierung des Wundalters zu. Da alpha-Aktin-positive Myofibroblasten im Untersuchungsgut auch noch in einer Hautwunde mit einem Alter von 2 Monaten und 13 Tagen beobachtet werden konnten, ist die im Tierexperiment gefundene maximale Nachweisbarkeitsdauer von 30 Tagen auf das Granulationsgewebe menschlicher Hautwunden nicht übertragbar.
    Notes: Summary Human skin wounds (66) inflicted between 20 h and 7 months prior to biopsy were studied. In order to identify the type of cellular differentiation of the fibroblastic cells in the granulation tissue, alpha-smooth muscle actin and desmin were immunohistochemically localized. The value of any presumed time-dependent appearance and/or disappearance of positively stained cells was tested for the estimation of wound age. In skin specimens with a wound age less than 5 days (n =15) no typical granulation tissue had developed and no alpha-actin-positive myofibroblasts could be detected. The first appearance of positively reacting myofibroblasts was noted in a 5-day-old wound. In 57% of the lesions with a wound age between 5 and 31 days (25 out of 44 cases) typical granulation tissue formation was present and myofibroblasts with positive reaction for alpha-smooth muscle actin could be identified. Numerous positively reacting cells could generally be found in wounds aged between 16 and 31 days, but also in wounds less than 16 days old. In 29% of the cases with a wound age of more than 31 days (2 out of 7 cases) alpha-sma-positive myofibroblasts also occured. Fibroblastic cells positive for desmin could not be seen at all in our series. Our results demonstrate the appearance of alpha-sma-positive myofibroblasts with the initial formation of typical granulation tissue in human skin lesions as early as approximately 5 days after wounding. In contrast to recent experimental results these cells remained detectable in wounds aged more than 2 months in some cases. The immunohistochemical detection of actin-positive cells, therefore, demonstrates whether an unknown skin wound is aged approximately 5 days or more. Even though a time-dependent decrease of myofibroblasts in human granulation tissue after 31 days in human wounds seems probable, the extended presence (up to about 2 months) of these cells allows no further exact age determination of older wounds.
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