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. 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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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.

     

Current role of scapulectomy

Tumors of the scapula are an unusual clinical challenge. Partial or complete resection of the scapula, with its attached musculoaponeurotic tissue, is a seldom used technique for the treatment of primary bone and soft tissue tumors, as well as selected metastatic involvement of the scapula. Scapulectomy may allow wide margins of resection without amputation. The purpose of this study is to review our recent experience with scapulectomy. This study describes the recent experience with scapulectomy by the Section of Surgical Oncology and the Department of Orthopedics at Louisiana State Medical Center (New Orleans, LA). Between 1994 and 1998, 12 patients (between 16 and 79 years of age) underwent a resection of the scapula. Eleven of these patients had soft tissue tumors; one had a metastasis from a thyroid carcinoma. Six of these patients underwent a scapulectomy as a primary treatment, five for recurrence. Six patients also received postoperative radiation and/or chemotherapy. The follow-up ranged from 6 months to 4 years. There was no mortality or wound infection associated with scapulectomy. All patients had normal hand and wrist function after surgery. Three distant recurrences occurred, with no local or regional failures encountered during the follow-up period. Scapulectomy can result in excellent local tumor control. Whereas some loss of active shoulder motion may occur, hand, wrist and elbow function is preserved. Although maintenance of shoulder function should not take precedence over adequacy of resection, scapulectomy remains an excellent procedure for malignant disease that preserves hand, wrist, and elbow function.     

Expression of intercellular adhesion molecule 1 (ICAM-1) is reduced in permanent focal cerebral ischemic mouse brain using an adenoviral vector to induce overexpression of interleukin-1 receptor antagonist

Our previous studies have demonstrated that overexpression of recombinant human interleukin-1 receptor antagonist protein (IL-1ra) via gene transfer can reduce ischemic brain injury. However, the mechanism of action of IL-1ra in ischemia is unclear. Since interleukin-1 can up-regulate intercellular adhesion molecules in endothelium, the present study was designed to determine whether overexpression of the IL-1ra can reduce the expression of intercellular adhesion molecule-1 (ICAM-1) after ischemic injury. Normal saline or adenovirus vector (1x109 particles) encoding the human IL-1ra gene (Ad.RSVIL-1ra) or the Escherichia coli LacZ gene (Ad.RSVlacZ) was injected into the right lateral cerebral ventricle of adult CD-1 mice. After five days, permanent middle cerebral artery occlusion (MCAO) was achieved for 24 h using an intraluminal suture. Cerebral blood flow was monitored by transcranial laser Doppler flowmetry to verify the occlusion. ICAM-1 protein was quantified using Western blot analysis and localized using immunohistochemistry. After MCAO, surface blood flow in the ischemic hemisphere was decreased to 9-11% of the baseline. There were fewer ICAM-1 positive vessels in the ischemic cortex of the Ad.RSVIL-1ra transfected mice than in the Ad.RSVlacZ transfected and saline treated mice (138+/-19 vs. 249+/-25, 284+/-22, p<0.05). Western blot analysis shows that ICAM-1 protein decreased 50-60% in the Ad. RSVIL-1ra group compared to the other two groups. There were no significant differences in the numbers of positive vessels in the ischemic basal ganglia and contralateral hemisphere among the three groups. Our studies suggest that IL-1ra overexpression can down-regulate the expression of ICAM-1 in the ipsilateral cortex in ischemic mice. Interleukin-1 may play an important role in the activation of inflammatory reaction during focal cerebral ischemia by promoting leukocyte adhesion on the endothelium cells.     

Lung cancer risk in male workers occupationally exposed to diesel motor emissions in Germany.

BACKGROUND: Although in several epidemiological studies exposure to diesel motor emissions (DME) shows an elevated lung cancer risk, it is still controversial whether DME is a human carcinogen. METHODS: In a pooled analysis of two case-control studies on lung cancer in Germany a total of 3498 male cases with histologically or cytologically ascertained lung cancer and 3541 male population controls were included. Information about lifelong occupational and smoking history was obtained by questionnaire. Drivers of lorries, buses, taxies, diesel locomotives and forklift trucks, bulldozers, graders, excavators, and tractors, were considered as exposed to DME and their cumulative exposure was estimated. All odds ratios were adjusted for smoking and asbestos exposure. RESULTS: The evaluation of lung cancer risk for all jobs with DME-exposure combined showed an odds ratio of OR=1.43 (95%-CI: 1.23-1.67). Most pronounced was the increase in lung cancer risk in heavy equipment operators (OR=2. 31 95%-CI: 1.44-3.70). The risk of tractor drivers increased with length of employment and reached statistical significance for exposures longer than 30 years (OR=6.81, 95%-CI: 1.17-39.51). The group of professional drivers (e.g., trucks, buses, and taxies), showed an increased risk only in West Germany (OR=1.44, 95%-CI: 1. 18-1.76), but not in East Germany (OR=0.83, 95%-CI: 0.60-1.14). DME-exposure in other traffic related jobs (e.g., diesel engine locomotive drivers, switchmen, forklift operators) was associated with an odds ratio of OR=1.53 (95%-CI: 1.04-2.24). CONCLUSIONS: The study provides further evidence that occupational exposure to diesel motor emissions is associated with an increased lung cancer risk.     

Primary vitrectomy without scleral buckling for rhegmatogenous retinal detachment

Background: Pars planta vitrectomy has evolved as an alternative method in the treatment of more complicated rhegmatogenous retinal detachments. We report a series of patients who underwent primary vitrectomy with gas tamponade without the use of additional scleral buckling. Methods: A retrospective study of 53 patients with a follow-up of 6–45 months (mean 17.8 months) was carried out. Preoperative findings included unusual, multiple or large breaks, vitreous haemorrhage, proliferative vitreoretinopathy and bullous retinal detachment. Preoperative visual acuity was between light perception and 1.0, with 30% (16/53) of patients with 0.4 or better. Results: Retinal reattachment was achieved in 64% of cases (34/53) with one and in 92% (49/53) with one or more operations. Final visual acuity was between light perception and 1.0, with 41% (22/53) of patients with 0.4 or better. Cataract formation occurred in 86% (37/43) of all patients with a clear lens preoperatively. Macular pucker was noted in 11 % (6/53) and postoperative proliferative vitreoretinopathy causing redetachment in 6% (3/53). Conclusion: With primary vitrectomy, a high final anatomical success rate with few intraoperative complications can be achieved in more complicated forms of rhegmatogenous retinal detachment. The major drawback of the procedure is the high incidence of post-operative cataract formation.