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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.

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.

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.

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.

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.

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.

Various congenital anomalies have been associated with childhood cancer, but as yet no anomaly has been consistently found with Ewing sarcoma (ES). Recently a large case-control study of ES patients reported a greater number of hernias in both cases and their sibling controls than in population controls. Most of these hernias were inguinal. Because these anomalies were also reported previously in two case series, we looked for inguinal hernias in a different population of ES patients.We abstracted medical records for 306 pathologically confirmed ES/primitive neuroectodermal tumor (PNET) patients seen at NIH between 1960 and 1992. Epidemiological data on demographics and medical conditions were analyzed. The frequency of anomalies was compared to expected rates to calculate relative risk and confidence intervals.Anomalies were present in 67 (22%) cases. A particular anomaly, inguinal hernia, was reported for 13 (5%) NIH cases. Compared to population estimates for white children, the relative risk of inguinal hernia among white NIH cases was 13.3 (95% CI 3.60-34.1) for females and 6.67 (95% CI 2.67-13.7) for males.The findings of inguinal hernias in some patients with ES suggest that a disruption in normal embryological development occurred. This may provide an important clue to the etiology of ES. We hypothesize that these hernias may relate to an in utero exposure or indicate an underlying genetic disorder. Future studies should carefully evaluate ES families for genetic disease and explore environmental factors. Med. Pediatr. Oncol. 34:195-199, 2000. Published 2000 Wiley-Liss, Inc.

The balanced reciprocal translocation t(11;22)(q24;q12) is a specific molecular marker for the Ewing's sarcoma family of tumors (ESFT). Based on the detection of this translocation in some olfactory neuroblastomas (ONBs), it has been proposed that ONBs also belong to the ESFT. Others have challenged this hypothesis; however, the rarity of ONBs, as well as the lack of molecular techniques that work reliably on formalin fixed tissue, have precluded the molecular analysis of a significant number of cases. We evaluated the immunophenotypic and molecular features of 17 paranasal small round neuroectodermal tumors using routinely fixed tissue. Probes localizing to 22q12 (EWS) and 11q24 (FLI-1) were used in a standard 2-color fluorescence in situ hybridization (FISH) assay to evaluate EWS/FLI1 fusion on the der(22). Sixteen tumors were mic-2 negative, whereas 1 tumor was mic-2 positive, compatible with ONB and ESFT, respectively. Thirteen of 15 ONBs could be evaluated by FISH, and all 13 were negative for the EWS/FLI1 fusion. Distinct fusion signals were identified in the single paranasal ESFT. Our findings indicate that ONBs lack the EWS/FLI1 fusion and are unrelated to the ESFT; however, true ESFTs may rarely occur as primary sinonasal tumors.

We previously reported that treatment of human peripheral blood monocytes or dendritic cells (DC) with calcium ionophore (CI) led to the rapid (18 hour) acquisition of many characteristics of mature DC, including CD83 expression. We therefore investigated whether less-mature myeloid cells were similarly susceptible to rapid CI activation. Although the promyelocytic leukemia line HL-60 was refractory to cytokine differentiation, CI treatment induced near-uniform overnight expression of CD83, CD80 (B7.1), and CD86 (B7. 2), as well as additional characteristics of mature DC. Several cytokines that alone had restricted impact on HL-60 could enhance CI-induced differentiation and resultant T-cell sensitizing capacity. In parallel studies, CD34(pos) cells cultured from normal donor bone marrow developed marked DC-like morphology after overnight treatment with either rhCD40L or CI, but only CI simultaneously induced upregulation of CD83, CD80, and CD86. This contrasted to peripheral blood monocytes, in which such upregulation could be induced with either CI or rhCD40L treatment. We conclude that normal and transformed myeloid cells at many stages of ontogeny possess the capacity to rapidly acquire many properties of mature DC in response to CI treatment. This apparent ability to respond to calcium mobilization, even when putative signal-transducing agents are inoperative, suggests strategies for implementing host antileukemic immune responses.

Asymmetry of the limbs (conventionally known as hemihypertrophy) is one of the overgrowth syndromes occurring sporadically in the general population at a frequency of approximately 1:86,000. Hemihypertrophy is also reported as part of the Beckwith-Wiedemann syndrome which has as its cardinal features omphalocele, macroglossia and gigantism with hypoglycemia, organomegaly, renal anomalies, hemihypertrophy, and embryonal tumors occurring less frequently. Various neoplasms are also associated with isolated hemihypertrophy. Wilms tumor, adrenocortical carcinoma, and hepatoblastoma are the most frequent. Rhabdomyosarcoma, neuroblastoma, phaeochromocytoma, and undifferentiated sarcoma of the lung are encountered only rarely. Loss of heterozygosity (LOH) of chromosome 11p15.5 is strongly associated with childhood embryonal tumors, particularly Wilms tumor, hepatoblastoma, and rhabdomyosarcoma.In this article, we describe an adolescent male with congenital asymmetry of the lower limbs who presented with a large poorly differentiated pelvic sarcoma. Conventional histologic, immunohistochemical, and ultrastructural studies of this tumor were insufficient for accurate subclassfication. However, positive staining for MyoD1 (a recently identified embryonically expressed marker of muscle differentiation) and LOH at the tyrosine hydroxylase locus of chromosome 11p15.5 by molecular analysis favored the diagnosis of embryonal rhabdomyosarcoma over an undifferentiated sarcoma.This case stresses the importance of pursuing clinical findings when they occur in conditions with an increased risk of developing cancer, which in this case was asymmetry of a limb. Also illustrated by this patient is the need for early consideration of molecular diagnostic tests where available, to refine an uncertain pathologic diagnosis that may ultimately have an impact on treatment and prognosis.

Analysis of human tumor cells in vitro enhances the study of numerous neoplastic conditions. However, it has been difficult to establish long-term cultures of adenoma cells, especially those of neuroendocrine origin, because the endocrine cells survive only briefly in culture, and fibroblasts overgrow the culture dish in 1 or 2 weeks. We describe cells isolated from pituitary adenomas in two patients with multiple endocrine neoplasia type 1 in which cells with a mesenchymal phenotype evolved from pituitary tumor cells. It appears that these poorly differentiated cells arose from multipotent adenoma cells. This represents a path of cell differentiation not observed previously in humans and may help explain the diverse nature of the benign tumors in multiple endocrine neoplasia type 1.

The insulin-like growth factors (IGF) I and II regulate metabolism, mitogenesis, differentiation, and apoptosis. The therapeutic uses of IGF-I have been discussed extensively; however, excessive activity of the IGF ligands and IGF-I receptor has been suggested as a factor in tumorigenesis. The inhibition of apoptosis by IGF-I is believed to be particularly important for the stimulation of tumor growth. This study examined whether systemic recombinant human IGF-I (rhIGF-I) therapy affects the growth of fibrosarcomas derived from fibroblasts expressing the IGF-I receptor at high or naturally occurring densities (1.9 x 10(5) compared with 1.6 x 10(4) IGF-I receptors/cell) in athymic nude mice. Treatment with 4 or 10 mg/kg rhIGF-I resulted in a marked reduction in the tumor latency and stimulated the growth of fibrosarcomas that overexpressed the IGF-I receptor. The latency and growth of fibrosarcomas expressing parental levels of the IGF-I receptor were not affected by rhIGF-I therapy. Analysis of mitosis by histone H3 mRNA in situ hybridization and of apoptosis by terminal deoxynucleotidyl transferase-mediated nick end labeling assay indicated that rhIGF-I-stimulated tumor growth was associated with a marked increase in mitogenesis; however, there was no evidence for any significant effect on apoptosis. These data imply that: (a) systemic rhIGF-I can stimulate the growth of tumors directly by stimulating mitosis; and (b) a reasonable level of IGF-I receptor expression is required for stimulation of tumor growth by systemic rhIGF-I.

We have previously shown that metastatic carcinomas of unknown primary site overexpress several tumor markers as well as the products of the oncogenes c-myc, ras and c-erbB2. We analyzed the tissue expression of the protein products of the apoptosis modulation genes p53 and bcl-2 in 47 CUP cases. Formalin-fixed, paraffin embedded tumor specimens were stained with commercially available antibodies to p53 (DO7) and bcl-2 after antigen retrieval by the microwave method. Staining was evaluated by intensity (1+ to 3+), percentage of positive cells (1-100%), and the 'intensity times percentage' product defined as the immunoreactivity index with values ranging from 0 to 300. Immunoreactivity index values higher than 150 were considered to characterize protein over-expression. Expression of p53 was identified in 70.2% of tumors while 53% of them showed a high immunoreactivity index. Bcl-2 expression was detected in 65% of tumors and overexpressed in 40%. Overexpression of both proteins was detected in 20% of tumors. The detection of either protein was not associated with any of the major clinicopathological variables studied. Nevertheless, a trend towards a more favourable response to platin based chemotherapy was seen in the cases that showed a strong expression of both proteins, when analysed by immunoreactivity index and percentage of positive cells. We conclude that CUP overexpress at a high percentage the p53 and the bcl2 proteins. The observed weak association of strong expression of these proteins with response to platin-based chemotherapy deserves further evaluation in the CUP setting.

Recent studies have demonstrated that telomerase, a reverse transcriptase linked to cellular "immortalization," is activated in a variety of malignant human tumors. This study was conducted to determine whether telomerase activity represents a marker of malignant transformation in precancerous (dysplastic) nodules arising in patients with cirrhosis.Telomerase activity was evaluated in frozen tissue samples of 14 cirrhotic liver specimens and 30 large nodular lesions contained therein, including 13 large regenerative nodules/low grade dysplastic nodules, 10 high grade dysplastic nodules, and 7 hepatocellular carcinomas (HCCs). A modified telomeric repeat amplification protocol was used.There was a clear-cut difference in telomerase activity levels between HCC (positive or strongly positive) and cirrhotic liver samples (weakly positive or negative). The majority of large noncancerous nodules (86%) exhibited telomerase activity levels similar to HCCs. However, such activity was not limited to dysplastic lesions but also was detected in some large regenerative nodules.These findings suggest that telomerase activation is an early event in large nodule formation in cirrhosis, which may facilitate the action of other factors in the process of carcinogenesis. Telomerase activity in large hepatic nodules is not always indicative of malignant transformation.

CRH, the principal regulator of the hypothalamic-pituitary-adrenal axis and modulator of autonomic nervous system activity, also participates in the regulation of appetite and energy expenditure. Antalarmin, a pyrrolopyrimidine compound, antagonizes CRH type 1 receptor-mediated effects of CRH, including pituitary ACTH release, stress behaviors, and acute inflammation. We administered antalarmin chronically to evaluate its effects on hypothalamic-pituitary-adrenal axis function and metabolic status. Adult male rats were treated twice daily with 20 mg/kg of i.p. antalarmin or placebo over 11 days. The animals were weighed; plasma ACTH, corticosterone, leptin, and blood glucose levels were determined; and morphometric analyses were performed to determine adrenal size and structure, including sizing, histochemistry, immunohistochemistry, and electron microscopy. Leptin messenger RNA expression in peripheral fat was analyzed by Northern blot. Antalarmin decreased plasma ACTH (mean +/- SD, 2.62 +/- 0.063 pg/ml) and corticosterone concentrations (10.21 +/- 1.80 microg/dl) compared with those in vehicle-treated rats [respectively, 5.3 +/- 2.0 (P0.05) and 57.02 +/- 8.86 (P0.01)]. Antalarmin had no significant effect on body weight, plasma leptin, or blood glucose concentrations or fat cell leptin messenger RNA levels. The width of the adrenal cortex of animals treated with antalarmin was reduced by 31% compared with that in controls without atrophy of the gland. On the ultrastructural level, adrenocortical cells were in a hypofunctional state characterized by reduced vascularization, increased content of lipid droplets, and tubulovesicular mitochondria with fewer inner membranes. The apoptotic rate was increased in the outer zona fasciculata of animals treated with the antagonist (26.6 +/- 3.58%) compared with that in placebo-treated controls (6.8 +/- 0.91%). We conclude that chronic administration of antalarmin does not affect body weight, carbohydrate metabolism, or leptin expression, whereas it reduces adrenocortical function mildly, without anatomical, clinical, or biochemical evidence of causing adrenal atrophy. These results are promising for future uses of such an antagonist in the clinic.