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Fatal injuries from firearms represent a small but important fraction of forensic autopsies. This article presents a retrospective medicolegal study of firearms-related deaths in Berlin from 2000 to 2009. The data analysis focused on the forensic and demographic characteristics and the relationship of these factors to each other. Furthermore, the survival time after gunshot injuries was also analyzed. In this study 332 deaths due to firearms wounds were analyzed of which 90 % were male and the mean age was 54.3 years. More than three quarters of the deaths were suicide mostly using illegally possessed weapons. The shooting hand could be identified in 59 % of the suicide cases. In 86 % the victim died at the scene or on the way to hospital. The median survival time was 2 h 27 min and 30 s. In cases where alcohol was consumed before the shooting incident, the majority (54 %) of victims were severely intoxicated (blood alcohol concentration > 1.5 mg/g). Handguns (pistols 53 % and revolvers 18 %) were most frequently used. Close range shots occurred in 81 %, shots with full perforation were most frequent (62 %) and on average 1.39 shots were fired. Most injuries (66 %) involved the head and the most common causes of death were perforating gunshot wounds to the head (55 %) and penetrating gunshot wounds (21 %). Most firearms-related suicides were committed in the summer months. Killing by firearms was found in 20 % and only one fatal shooting accident was found during the study period. Firearms-related deaths continue to pose a relevant problem in terms of suicide and illegally possessed weapons. Each case requires an interdisciplinary approach as individual morphological aspects cannot per se be considered as evidential for homicidal, suicidal or accidental death. The number of gunshot wounds does not necessarily influence the survival time. Language: de

Hintergrund Verletzungen nach einem Sturz aus einer Hohe > 3 m stellen haufige Ursachen eines Polytraumas und zu untersuchende Todesursachen in der forensischen Obduktionsroutine dar.

Previous references suggesting a high mortality of propofol addiction in medical personnel were mostly based on surveys of the heads of medical departments or case reports; therefore, a questionnaire was sent to 48 forensic medicine departments in Germany, Austria and Switzerland concerning the number of autopsies carried out between 2002-2112 on medical personnel with the suspicion of abuse of propofol or other analgesics. The response rate was 67%. In 16 out of the 32 responding departments 39 deaths (27 males) were observed with previous connections to anesthesiology, intensive care or emergency departments of which 22 were physicians, 13 nurses, 2 other personnel and 2 were unknown. Propofol was the major cause of death in 33 cases (85%), in 8 cases including 7 with propofol, an unintentional accident was recorded and 29 were determined to be suicide. In 14 cases chronic abuse was denied but actually excluded by toxicological analysis in only 2 cases. In 11 cases involving suicide the question of abuse was not investigated. This survey confirmed previous data about the central role of propofol for the fatal outcome of addiction and suicide of anesthetists and other medical personnel. A dual prevention strategy with low-threshold offers for persons at risk and strategies for early detection is urgently needed including a stricter control of dispensing, improvement in forensic medical documentation and the use of toxicological investigations in every case of suspected abuse.

There are considerable similarities and intersections between forensic medicine and emergency medicine. This applies especially to frustraneously resuscitated patients or other lethal clinical courses of traumatized patients who are subject to latter forensic autopsy. Cooperation between departments of emergency and forensic medicine not only has emergency medical training potential, but also the possibility of retrospective evaluation of medical emergency measures - both in individual cases and with regard to epidemiological aspects. In particular, the widespread registration of autopsied pre-hospital trauma deaths that occurred despite on-scene resuscitation attempts is useful. The pre-hospital situation represents a hotspot, but also a blind spot in the overall trauma mortality. In recent clinical registers, preclinical deaths go mostly unrecorded, despite the undisputed benefits of clinical registers.

Es wird der plotzliche Todesfall einer 20 Jahre alt gewordenen Frau vorgestellt, bei der sich autoptisch eine homogene Schwarzfarbung beider Schilddrusenlappen zeigte. Histologisch fanden sich reichlich dunkelbraunlich-schwarzliches Pigment im Zytoplasma der Follikelepithelien und vereinzelt Pigment auch im Kolloid in den Lichtungen der Schilddrusenfollikel. Diese schwarzliche Verfarbung beider Schilddrusenlappen, auch als „black thyroid“ bezeichnet, ist ein seltener Befund ohne Krankheitswert, der vornehmlich auf die Einnahme des Antibiotikums Minocyclin zuruckgefuhrt wird. Bislang sind weltweit weniger als 100 Falle in der Literatur beschrieben worden.

Fatal gunshot injuries with unusual firearms are rare. A total of 16 cases of suicide by self-constructed or manipulated firearms from the years 2007-2009 are described. Technical characteristics of the weapons involved as well as morphological findings and incident reconstructions are presented. In addition a review of the literature is given. The weapons involved in this study were 6 self-constructed shooting devices, 5 manipulated blank firearms, 2 unmodified blank firearms, 1 signal pistol for maritime usage, 1 sawn-off shotgun and 1 replicate of a muzzle loader. The efficiency of the weapons involved achieved by manipulation or by construction is shown in this study as well as indications for the medicolegal expertise in such cases.

Resuscitation attempts require invasive iatrogenic manipulations on the patient. On the one hand, these measures are essential for successful survival of the patient, but on the other hand can damage the patient. We differentiate between frequent and rare injuries. Factors of influence are duration and intensity of the resuscitation attempts, age of the patient as well as a coagulate-inhibiting medication.

Maternal deaths during pregnancy, both from pregnancy-related and other causes, are rare in Western industrialized countries. Pregnancy-induced hypertension and amniotic fluid embolism (AFE) are the main contributors to directly gestation-related maternal death in pregnancy. The forensic evaluation of such a case should include autopsy, clinical details, and microscopic and toxicological investigations, noting that autopsy findings are often nonspecific. Even if the cause of death is apparently obvious after autopsy, histopathological examination should be performed in suspected pregnancy-related fatalities. In cases of suspected AFE, representative histological specimens from different areas of each organ, in particular from pulmonary tissue, should be collected and examined.

Tension pneumothorax can occur at any time during cardiopulmonary resuscitation (CPR) with external cardiac massage and invasive ventilation either from primary or iatrogenic rib fractures with concomitant pleural or parenchymal injury. Airway injury can also cause tension pneumothorax during CPR. This article presents the case of a 41-year-old woman who suffered cardiopulmonary arrest after undergoing elective mandibular surgery. During CPR the upper airway could not be secured by orotracheal intubation due to massive craniofacial soft tissue swelling. A surgical airway was established with obviously unrecognized iatrogenic tracheal perforation and subsequent development of tension pneumomediastinum and tension pneumothorax during ventilation. Neither the tension pneumomediastinum nor the tension pneumothorax were decompressed and accordingly resuscitation efforts remained unsuccessful. This case illustrates the need for a structured approach to resuscitate patients with ventilation problems regarding decompression of tension pneumomediastinum and/or tension pneumothorax during CPR.

Eine 52-jahrige Frau wurde tot in ihrer Wohnung aufgefunden. Bereits bei der auseren Leichenschau fielen zahlreiche grosere und kleinere Hamatome am gesamten Korper auf, sodass eine gerichtliche Sektion angeordnet wurde, um ein Gewaltverbrechen auszuschliesen. Die Obduktion zeigte eine massive Blutgerinnungsstorung (Verbrauchskoagulopathie). Todesursachlich war eine Hirnblutung unklarer Genese. Wegen einer eventuellen infektionsbedingten Eigen- oder Fremdgefahrdung und dem davon abhangigen weiteren Vorgehen mussten u. a. moglichst viele Differenzialdiagnosen hamorrhagischer Ereignisse zeitnah ausgeschlossen werden. Die endgultige Diagnose einer akuten myeloischen Leukamie konnte erst histologisch gestellt werden. Diese Kasuistik zeigt, dass es sich bei der akuten myeloischen Leukamie um eine heterologe Erkrankung handelt, die u. a. differenzialdiagnostisch als Ursache eines plotzlichen Todes in Betracht gezogen werden muss.

Bei Simon-Blutungen handelt es sich um Unterblutungen des vorderen Langsbandes der Wirbelsaule vor den Zwischenwirbelscheiben. Simon-Blutungen sind ausfuhrlich bei Fallen von Tod durch Erhangen beschrieben worden und treten hier vorwiegend im Bereich der Lendenwirbelsaule auf. In der Literatur wurde uber Simon-Blutungen auch in einigen Fallen von Ertrinken, bei verschiedenen traumatischen Todesursachen und uberdurchschnittlich haufig bei Faulnisveranderungen berichtet. In einer prospektiven Untersuchung von 600 konsekutiven Sektionsfallen, die unabhangig von der Todesursache durchgefuhrt wurde, wurden in 14 von insgesamt 20 Erhangungsfallen Simon-Blutungen festgestellt. Ebenso fanden sich diese auch in 5 Fallen anderer Todesursachen. Hierbei handelte sich je um einen Tod in aufrechter Korperposition, eine Drogenintoxikation, eine Lungenembolie, einen Herzinfarkt und einen Sturz. Anhand dieser Kasuistiken werden differenzialdiagnostische Uberlegungen zur Entstehung und zur Bedeutung der Simon-Blutungen bei anderen Todesursachen als Erhangen vorgestellt. Ferner zeigte sich, dass bei fortgeschrittener Faulnis echte Unterblutungen des Langsbandes auch histologisch nicht von einer Durchtrankung mit hamolytischem Blut bzw. Faulnisflussigkeit abgegrenzt werden konnen.

Es wird uber einen 79-jahrigen Patienten berichtet, der einige Stunden nach Katheterablation zur Therapie von Vorhofflimmern unter dem klinischen Bild einer Herzbeuteltamponade verstarb. Obduktion und nachfolgende mikroskopische Untersuchungen ergaben den Nachweis elektrothermischer Nekrosen in beiden Vorhofen mit kompletter Vorhofwandperforation rechts im Bereich einer Koagulationsstelle. Postmortal erhobene, ablationsbezogene Befunde (Koagulationsnekrosen von Muskulatur, Gefaswand) werden dargestellt und diskutiert sowie eine autoptische Vorgehensweise empfohlen. Es erfolgt eine Einschatzung des Nutzen-Risiko-Verhaltnisses der – zu befurwortenden – therapeutischen elektrophysiologischen bzw. elektroablativen Untersuchungen.

Ein 79-jahriger Patient verstarb dreieinhalb Stunden nach Durchfuhrung einer Katheterablation zur Therapie von Vorflimmern unter dem klinischen Bild einer Herzbeuteltamponade. Obduktion und mikroskopische Untersuchungen ergaben den Nachweis elektrothermischer Nekrosen in regularer topographischer Zuordnung zu den Pulmonalvenen linksatrial bzw. zum rechtsatrialen Isthmus mit kompletter Vorhofwandperfortion rechts im Bereich einer Koagulationsstelle. Postmortal erhobene, ablationsbezogene Befunde werden dargestellt und diskutiert sowie mogliche Ursachen der zeitlich verzogerten Herzbeuteltamponade erortert.

Die Pyomyositis ist eine eitrige Entzundung der Skelettmuskulatur, die uberwiegend durch Staphylococcus aureus verursacht wird. Je nachdem, welche Muskelgruppen involviert sind, kann die Pyomyositis diverse Erkrankungen imitieren und durch eine verzogerte Diagnosestellung todlich verlaufen. Durch die Moglichkeit einer gezielten Praparation der Muskulatur lasst sich die Pyomyositis autoptisch in der Regel ohne Schwierigkeiten nachweisen. In 40% der Falle bestehen multiple intramuskulare Abszesse. Im Anfangsstadium der Erkrankung zeigt sich histologisch lediglich ein Odem der Muskulatur. In der Folge wird die betroffene Muskulatur von massenweise Entzundungszellen durchsetzt, die die Muskelfasern auseinander drangen. Nekrosen der Muskelfasern mit Verlust der Querstreifung, einhergehend mit einer Faserhomogenisierung und zerfallenen, nekrotischen Muskelfaserfragmenten sowie das Auftreten einer pathologischen Langsstreifung der Muskelfasern sind in jedem Stadium der Pyomyositis nachweisbar. Eine verspatete Diagnosestellung fuhrt durch septische Komplikationen in 10% der Falle zum Tod der Betroffenen. Bei autoptisch nachgewiesener Pyomyositis mit todlichem Ausgang liegt die rechtsmedizinische Bedeutung der Erkrankung in dem Umstand, dass eine vorangegangene Traumatisierung der Muskulatur (z. B. Sturz, Schlag, Verkehrsunfall) ursachlich fur das Anlaufen der Erkrankung gewesen sein kann. Bei von fremder Hand beigebrachtem Trauma kommt z. B. eine Korperverletzung mit Todesfolge oder auch eine fahrlassige Totung in Betracht.

Die haufigsten Erreger der bakteriellen Meningitis sind in Deutschland Neisseria meningitidis, Streptococcus pneumoniae, Borrelia burgdorferi, Listeria monocytogenes und Staphylokokken. Nachdem die Impfung gegen Haemophilus influenzae in die Routineimpfprogramme aufgenommen wurde, spielt dieser Erreger im Gegensatz zu fruher kaum noch eine Rolle als Meningitiserreger. Eine Anzahl klinischer Pilotstudien deutet darauf hin, dass eine universelle Breitband-Polymerase-Kettenreaktion (Breitband-PCR) mit anschliesender speziesspezifischer PCR die Zukunft der atiologischen Diagnostik der bakteriellen Meningitis darstellen konnte. Die einfachste und schnellste diagnostische Methode stellt, nach wie vor, das Gram-Praparat dar. Bei perakut todlich verlaufenden Fallen, speziell beim Waterhouse-Friderichsen-Syndrom, lasst sich autoptisch allenfalls eine vermehrte Gefaszeichnung der Hirnoberflache nachweisen. Bei diesen Fallen findet sich histologisch z. T. noch uberhaupt keine Enzundungsreaktion der Leptomeninx. Bei der eitrigen Meningitis ist nach mehrtagigem Krankheitsverlauf neben der typischen Infiltration des Subarachnoidalraums mit massenhaft Granulozyten ein vielfaltiges histomorphologisches Bild pathologischer Veranderungen mit fibrinoiden Gefaswandnekrosen, Thrombosierungen, Ventrikulitis, Infarzierungen sowie venosen und arteriellen Vaskulitiden anzutreffen. Die Storung der Integritat der Blut-Hirn-Schranke bei bakterieller Meningitis beruht offenbar im Wesentlichen auf einer Separation der interzellularen „tight junctions“ des Endothels der Leptomeninxkapillaren. Todesursachlich ist, auch in Abhangigkeit von Schwere und Dauer einer Sepsis, eine Erhohung des intrakraniellen Drucks. Diese fuhrt in einem Circulus vitiosus (uber eine verminderte zentrale Perfusion, einhergehend mit metabolischer Azidose) zu einer zerebralen Vasodilatation. Hierdurch kommt es zu einer weiteren Hirndruckzunahme und letztlich zu zerebraler Minderperfusion und zentraler Dysregulation. Vor dem Hintergrund einer moglichen Verkennung der Erkrankung bei unzureichender arztlicher Diagnostik oder einer verzogerten Diagnosestellung sowie unter der Fragestellung posttraumatischer oder nosokomialer Genese der Erkrankung muss sich der rechtsmedizinische Gutachter gelegentlich mit der Thematik auseinander setzen.

Wir berichten uber eine nichttraumatische intraperitoneale Ruptur der Harnblase bei einem 74-jahrigen Mann. Am 8. postoperativen Tag nach transurethraler Resektion der Prostata (TUR-P) trat eine massive, diffuse Parenchymblutung aus arteriellen Gefasen der Prostataloge auf. Uber die Ausbildung einer Harnblasentamponade mit Spontanruptur kam es dann zu Peritonitis, Sepsis und Tod. Eine traumatische Verletzung der Blase im Rahmen der TUR-P konnte aufgrund des unauffalligen klinischen Verlaufes bis zum 8. postoperativen Tag und dem Umstand, dass die postoperative Kontrastmitteldarstellung der Blase und Resektionsloge paravasatfreie Verhaltnisse zeigte, ausgeschlossen werden. Das Vorliegen von Faktoren, die fur eine spontane Harnblasenruptur pradisponieren, wie z. B. Zystitis, Tumor, Divertikel oder Bestrahlungsfolgen, konnte durch die Obduktion bzw. histologische Untersuchung ausgeschlossen werden. Eine Spontanruptur der Harnblase nach Blasentamponade ist unseres Wissens in der Literatur bisher nicht beschrieben worden. Auch finden sich keine Mitteilungen uber das Auftreten einer Harnblasentamponade als Folge von Nachblutungen nach TUR-P.

Es wird uber eine 77-jahrige Frau, die mit dem Rezidiv eines Wurzelkompressionssyndroms bei einem Bandscheibenvorfall stationar aufgenommen wurde und 2 Tage spater verstarb, berichtet. Sektion und Histologie ergaben die Befunde einer von einer eitrig-abszedierenden Bronchopneumonie ausgehenden bakteriellen Sepsis, mit septikopyamischer Streuung in Herz und Gehirn. Daruber hinaus zeigte sich eine Lungenfettembolie II. bis uberwiegend III. Grades. Fettembolien geringeren Ausmases waren auch in anderen Organen nachweisbar. Die Pathogenese der Fettembolie wird vor dem Hintergrund der Literaturangaben erortert. Die Gesamtkonstellation spricht fur eine hepatogene Fettembolie bei vorbestehender, teilweise zystischer Leberverfettung, mit Freisetzung und Embolisierung des Fettes aus stauungsbedingten Parenchymnekrosen bei zunehmender Rechtsherzinsuffizienz. Die Lungenfettembolie war klinisch relevant und mitursachlich fur den Tod.

Der vorliegende Fallbericht beschreibt den trotz Intensivtherapie letalen Verlauf einer nach Alkoholexzess ambulant erworbenen Aspirationspneumonie bei einer 25jahrigen Patientin. Die histologischen Lungenbefunde zeigen ein ungewohnlich schweres Bild einer fulminant verlaufenen Aspirationspneumonie mit ausgedehnten peptischen Nekrosen der Alveolarwande und der Bronchialschleimhaut, hamorrhagischen Infarkten, bis nach intraalveolar reichenden Aspirationsherden und hyalinen Membranen. Es besteht eine direkte Relation zwischen der Hohe des pH-Wertes des Magensaftes und der Letalitatsrate von Aspirationspneumonien: je niedriger der pH-Wert, desto gravierender die Destruktion des Lungenparenchyms und umso hoher die Letalitat. Die Prognose alkoholtoxischer Aspirationspneumonien wird somit wesentlich durch die Hohe der Blutalkoholkonzentration (BAK) bestimmt, da Ethanol den pH-Wert des Magensaftes uber eine vermehrte Gastrinfreisetzung absenkt und damit im Falle einer Aspiration das Ausmas der peptischen Nekrosen der Alveolarwande und Bronchialschleimhaut festlegt und die Schwere einer moglicherweise letalen respiratorischen Insuffizienz definiert.

Resuscitation attempts require invasive iatrogenic manipulations on the patient. On the one hand, these measures are essential for successful survival of the patient, but on the other hand can damage the patient. We differentiate between frequent and rare injuries. Factors of influence are duration and intensity of the resuscitation attempts, age of the patient as well as a coagulate-inhibiting medication.

This article analyses three cases of death following a spontaneous rupture of the urinary bladder. One case is based on an extensive tamponade of the bladder eight days after a transurethral resection of the prostate gland. Two other cases of death by spontaneous rupture resulted from increased alcohol consumption. The paper presents an overview of pathomorphological findings such as the typical intraperitoneal rupture localisation at the posterior wall of the urinary bladder or bladder dome and the subsequent diagnosed causes of death. In addition predisposing (anamnestic) influencing factors such as chronic alcoholism are highlighted and their relevance for the clinical urologist and the forensic pathologist are discussed.

A 79-year-old man died 3.5 hours after catheter ablation of atrial fibrillation due to pericardial tamponade. Autopsy and consecutive microscopic examinations furnished evidence of electrothermical necroses topographically arranged regularly adjacent to the pulmonary veins in the left atrium and across the area of the isthmus in the right atrium with a rupture of the right atrial wall in the area of a coagulation spot. The post-mortem findings and pathogenetic considerations for the delayed pericardial tamponade are discussed.

Two cases of planned complex suicide are presented. In both cases, the suicidal persons shot themselves after making sure that in the sequel their bodies would be burnt. In the first case, a 65-year-old man had shot himself in the mouth and consequently fell into a fire he had lighted before. In the second case, a 43-year-old man set fire to his flat and shot himself directly afterwards. On the basis of the reported cases, a short literature review on planned complex suicides is given.

6 cases of self-inflicted injuries in male individuals are reported. The age of the affected men was between 15 and 46 years whereas the younger age predominated. Alleged incidents were robberies in 3 cases, rape in one case, violation in custody in one case and an assault originating in personal motives in one case. In 4 cases, the typical injury pattern of self-infliction was present showing parallel course and superficiality of the wounds in areas accessable by the persons's own hands. In 2 cases, atypical injuries (i.e. deep cuts and massive signs of strangulation respectively) were found. However, in most cases, the underlying motive was to gain affection. An autoerotic accident was tried to be disguised in one case. Profit was the leading motive in another case.

Of the side effects occurring in temporal association with the use of nonsteroidal anti-inflammatory drugs (NSAIDs), peptic ulcer disease is reported most often.To (1) provide information on the temporal association between fatal peptic ulcer presenting as sudden death and NSAID use prior to death, and (2) to examine the diagnostic efficiency of postmortem determination of NSAID levels using high-pressure liquid chromatography.Prospective autopsy study of all cases of sudden death associated with peptic ulcer disease from a total of 1139 medicolegal autopsies performed during a 12-month period.Postmortem femoral blood samples were analyzed for NSAIDs using high-pressure liquid chromatography, and specimens of gastric and duodenal mucosa were examined for coexisting pathologic conditions.Twelve fatalities that occurred out of hospital as a result of peptic ulcer disease and presented as sudden death were identified. Autopsy blood samples were positive for NSAIDs in 7 cases (ibuprofen in 4 cases, levels 0.8 to 1.4 microg/mL; diclofenac in 2 cases, levels 0.6 and 1.6 microg/mL; and ketoprofen in 1 case, level 0.3 microg/mL). The ages of the affected individuals (3 men, 4 women) ranged from 43 to 60 years. No other drugs, including corticosteroids, anticoagulants, salicylic acid, and salicylates, were present. Microscopic examination revealed no pathologic antemortem mucosal conditions in any of the cases.For the postmortem elucidation of etiopathogenetic factors contributing to fatal peptic ulcer disease, high-pressure liquid chromatography to determine NSAID levels in autopsy blood samples is of considerable diagnostic benefit, especially when combined with histology. The number of cases of sudden death involving younger individuals dying as a result of peptic ulcer disease in temporal association with preceding use of NSAIDs seems to be underestimated from the clinical viewpoint due to the underrepresentation of out-of-hospital fatalities in the field of clinical pathology.

Suicidal electrocution using a time switch is relatively rare. Based on two cases from the authors' own forensic material and seven cases from literature peculiarities regarding the concomitant intake of medicines (benzodiazepines) and alcohol are discussed and comments are made how to proceed in the criminal and forensic investigation of such cases.

A 70-year-old farmer was found dead in front of a barn. The emergency doctor did not note any significant findings at the external examination of the body. When the corpse was picked up by the undertaker, he noticed facial injuries. Several domestic cats were strolling around the barn at this time. External examination by a forensic pathologist revealed both sharp and crenated wound margins around the removed ear with punctured stabs adjacent to the mutilation. Further small perforating but not mutilating skin lesions were found also on other parts of the face. Given the typical morphology of the skin lesions and the observation of cats at the death scene, it is concluded that the injuries were inflicted by at least one domestic cat. In the forensic literature, reports on depredation of human corpses by cats are rare. To our knowledge, this is the first case report of outdoor cat depredation in the early postmortem interval. The case shows that domestic cats feed on human corpse even if other food is available.

Fas-mediated apoptosis proceeds though mitochondria-dependent or -independent pathways and is deficient in drug-resistant cells. Neuroblastoma, a common pediatric malignancy, often develops drug-resistance and has a silenced caspase 8 (FLICE) gene, which has been associated with Fas- and drug-resistance. We report that besides caspase 8, which was absent in approximately one-third of 26 neuroblastoma cases in this study, other proteins such as bcl-2 and FLICE-inhibitory protein (FLIP), are equally important in conferring Fas-resistance to neuroblastoma cells. Both bcl-2 and FLIP were frequently expressed in neuroblastoma tissues. Our in vitro studies showed that FLIP was recruited to the death-inducing signaling complex and interfered with the recruitment of caspase 8 in neuroblastoma cells. bcl-2 inhibited the activation of the mitochondria; but it also lowered the free cytoplasmic levels of caspase 8 by binding and sequestering it, thus acting through a novel antiapoptotic mechanism upstream of the mitochondria. In vitro down-regulation of bcl-2 with antisense oligonucleotides allowed the release of cytochrome c from mitochondria and the activation of caspases 8 and 3 upon Fas activation as well as sensitized neuroblastoma cells to Fas-mediated apoptosis. Down-regulation of FLIP had only a modest apoptotic effect because of the coexistent mitochondrial block. However, combined treatment with bcl-2 and FLIP antisense oligonucleotides had a statistically significant synergistic effect reversing Fas-resistance in neuroblastoma cells in vitro. These data indicate that Fas-mediated apoptosis in neuroblastoma cells is mitochondria-dependent and inhibited both at the mitochondrial level and at the level of caspase 8 activation. Thus, gene-targeting therapies for bcl-2 and FLIP may reverse Fas-resistance and prove useful in the treatment of drug-resistant neuroblastomas.

Problems with intubation of the ampulla Vateri during diagnostic and therapeutic endoscopic maneuvers are a well-known feature. The ampulla Vateri was analyzed three-dimensionally to determine whether these difficulties have a structural background.Thirty-five human greater duodenal papillae were examined by light and scanning electron microscopy as well as immunohistochemically.Histologically, highly vascularized finger-like mucosal folds project far into the lumen of the ampulla Vateri. The excretory ducts of seromucous glands containing many lysozyme-secreting Paneth cells open close to the base of the mucosal folds. Scanning electron microscopy revealed large mucosal folds inside the ampulla that continued into the pancreatic and bile duct, comparable to valves arranged in a row.Mucosal folds form pocket-like valves in the lumen of the ampulla Vateri. They allow a unidirectional flow of secretions into the duodenum and prevent reflux from the duodenum into the ampulla Vateri. Subepithelial mucous gland secretions functionally clean the valvular crypts and protect the epithelium. The arrangement of pocket-like mucosal folds may explain endoscopic difficulties experienced when attempting to penetrate the papilla of Vater during endoscopic retrograde cholangiopancreaticographic procedures.

To provide an investigative tool for the study of osteosarcoma (OSA) biology we have developed a syngeneic (balb/c) murine model of OSA, using cell lines derived from a spontaneously occurring murine OSA (Schmidt et al. Differentiation 1988; 39: 151-60). This model is characterized by orthotopic primary tumor growth, a period of minimal residual disease, spontaneous pulmonary metastasis, and clonally related variants (K7M2 and K12) that differ in pulmonary metastatic potential. Primary tumor and pulmonary metastasis histology was consistent with OSA in human patients. Expression of bone sialoprotein, biglyan, decorrin, and osteopontin was suggestive of bone lineage cells. The development and use of a more aggressive OSA cell line (K7M2) resulted in spontaneous metastasis to the lungs in over 90% of mice, whereas metastases were seen in only 33% of mice when a less aggressive OSA cell line (K12; Schmidt et al. Differentiation 1988; 39: 151-60) was used. Death from metastasis occurred at a median of 76 days using K7M2 whereas no median was achieved after 140 days using K12. Angiogenic potential, characterized by CD31 and factor VIII staining of primary tumors and pulmonary metastases, was greater in the K7M2 model compared to the K12 model. No significant differences in the in vitro or in vivo expression of angiogenesis associated genes (flt1, flt4, TIE1, TIE2, and VEGF) was found between K7M2 and K12. This well characterized and relevant model of OSA will be a valuable resource to improve our understanding of the biology and treatment of metastasis in OSA.

In this study, we investigated the sensitivity of Ewing's sarcoma family tumors (ESFTs) of children and adolescents to the tumor necrosis factor-related apoptosis-inducing Ligand (TRAIL). TRAIL binds to death receptors (DRs) DR4, DR5, DcR1, and DcR2. Either DR4 or DR5 can induce apoptosis, whereas DcR1 and DcR2 are considered inhibitory receptors. Nine of 10 ESFT cell lines, including several that were Fas resistant, underwent apoptosis with TRAIL through activation of caspase-10, capase-8 (FLICE), caspase-3, and caspase-9. In contrast to the Fas signaling pathway, caspase-10, but not caspase-8 or the Fas-associated death domain-containing molecule, was recruited to the TRAIL receptor-associated signaling complex. We found that 9 of 10 ESFT cell lines expressed both DR4 and DR5 by Western blotting, whereas the TRAIL-resistant line expressed only DR4. However, DR4 was absent from the cell surface in the resistant and two additional lines (three of five tested lines), suggesting that it may have been nonfunctional. On the contrary, DR5 was located on the cell surface in all four sensitive lines tested, being absent only from the cell surface of the resistant line that was also DR5-negative by Western blotting. In agreement with these findings, the resistance of the line was overcome by restoration of DR5 levels by transfection. Levels of DcR1 and DcR2 or levels of the FLICE-inhibitory protein (FLIP) did not correlate with TRAIL resistance, and protein synthesis inhibition did not sensitize the TRAIL-resistant line to TRAIL. Because these data suggested that sensitivity of ESFTs to TRAIL was mainly based on the presence of DR4/DR5, we investigated the presence of these receptors in 32 ESFT tissue sections by immunohistochemistry. We found that 23 of 32 tumor tissues (72%) expressed both receptors, 8 of 32 (25%) expressed one receptor only, and 1 was negative for both. Our finding of wide expression of DR4/DR5 in ESFT in vivo, in combination with their high sensitivity to TRAIL in vitro and the reported lack of toxicity of TRAIL in mice and monkeys, suggests that TRAIL may be a novel effective agent in the treatment of ESFTs.

Recent evidence suggests that one mechanism whereby cytotoxic drugs, such as doxorubicin, kill tumors is the induction or up-regulation of Fas ligand (FasL) expression on the tumor cell surface. The ensuing engagement of Fas by FasL on adjacent cells leads to apoptosis. However, despite cytotoxic drug-induced FasL expression, Fas-sensitive tumors frequently resist chemotherapy, suggesting that they may possess a mechanism that prevents or inactivates Fas-FasL interactions. In the present work, we addressed the involvement of the FasL/Fas signaling pathway in doxorubicin-induced apoptosis and the ability of matrix metalloproteinases (MMPs) to proteolytically cleave FasL in tumor cells. Doxorubicin-induced apoptosis was inhibited by expression of soluble Fas or incubation of the tumor cells with MMP-7 but not with MMP-2 or MMP-9. Resistance to doxorubicin was also induced by expression in the tumor cells of constitutively active MMP-7 but not of a catalytically inactive mutant. Conversely, inhibition of MMP-7 expression in tumor cells by transfection of MMP-7 cDNA in antisense orientation resulted in sensitization to doxorubicin. MMP-7 efficiently cleaved recombinant FasL in vitro and reduced cell surface FasL expression. Our observations provide evidence that one mechanism whereby MMP-7 may promote tumor survival and resistance to doxorubicin is by cleaving FasL and reducing its effectiveness in triggering Fas-mediated apoptosis.