Werner Jacobs

Syndecan‐1 expression in malignant mesothelioma: correlation with cell differentiation, WT1 expression, and clinical outcome

Syndecan‐1 binds basic fibroblast growth factor (bFGF), modulates neovascularization, plays a role in epithelial differentiation and is up‐regulated by WT1. Malignant mesothelioma of the pleura is one of the most aggressive tumours known and expresses high levels of angiogenic growth factors. This study has analysed syndecan‐1 expression in mesothelioma tumours and cell lines by immunohistochemistry and immunoblotting, using anti‐syndecan‐1 antibody directed against the core protein, and has examined its relation to morphology, bFGF, WT1, and intra‐tumoural microvascular density (IMD). Shedding of syndecan‐1 in the conditioned medium of mesothelioma cell lines was detected in variable amounts. These studies indicate that (1) there is no correlation of syndecan‐1 with either bFGF expression or IMD in mesotheliomas in vivo; (2) syndecan‐1 is strongly expressed in the epithelial type of mesothelioma and in the epithelial component of biphasic mesotheliomas and the expression is reduced or lost in sarcomatoid differentiation; together with the finding that (3) syndecan‐1 correlates with WT1 immuno‐expression, this suggests that syndecan‐1 might relate to the differentiation state of mesothelial/mesothelioma cells; and (4) syndecan‐1‐positive tumours are associated with a longer survival (p =0·02) than mesotheliomas with no or little syndecan‐1 expression, on univariate analysis. These findings therefore indicate that syndecan‐1 can be an important prognostic indicator in mesotheliomas and its loss may be important in the epithelial–mesenchymal transformation of mesothelioma cells. Copyright

Schistosoma mansoni infection causing diffuse enteric inflammation and damage of the enteric nervous system in the mouse small intestine.

Schistosomiasis mansoni is a major health problem, mainly occurring in developing countries. A large proportion of infected individuals suffers from motility‐related gastrointestinal problems. In the present study, the diffuse inflammatory response in the small bowel wall, as compared to the egg‐induced granulomatous inflammation, was investigated. For this purpose, OF1 mice infected with Schistosoma mansoni 8–16 weeks prior to the experiment, and uninfected control mice were studied. The ileum showed both a diffuse mucosal inflammation as well as a granulomatous reaction. The diffuse mucosal inflammation caused an increase in the thickness of the mucosa, with blunting of the villi. A significant, transient increase of thickness of the muscularis propria after 12 weeks of infection was noted. There was an infection‐related mast cell infiltrate in the muscularis propria, consisting of formalin fixation‐insensitive connective tissue mast cells. Ganglionitis of the myenteric plexus was noted. Rarely, ganglia of the myenteric plexus contained apoptotic cells. A general pharmacological set of experiments showed a significant increase in intestinal contractility, both to exogenously administered, as well as to endogenously released neurotransmitters. Our results demonstrate that S. mansoni infection in the mouse ileum leads to diffuse specific enteric inflammation that is associated with an enhanced response to contractile agents.

Schistosomal granuloma modulation. II. Specific immunogenic carbohydrates can modulate schistosome-egg-antigen-induced hepatic granuloma formation.

Abstract To further investigate the factors involved in the modulation of the schistosomal granuloma, mice were primed with immunogenic carbohydrates which were common to soluble egg antigen (SEA) and adult worm antigen. Mice sensitized with LewisX trisaccharide or lacto-N-fucopentaose-III (LNFP-III) displayed an increased cellular response towards SEA-coupled beads implanted in the liver by mesenteric injection, resulting in the formation of larger periparticular granulomas. When animals were sensitized with bovine serum albumin or a structurally related carbohydrate, an accelerated response was not seen. Since LNFP-III is built up of LewisX molecules, and LewisX carbohydrates are common to SEA and worm antigens such as the gut-secreted antigens CCA and CAA (two antigens that could prime egg-antigen-induced granuloma formation), this may explain why adult, live Schistosoma mansoni worms positively modulate egg-antigen-induced hepatic granuloma formation in the murine host. These observations provide new insights into the role of carbohydrates in parasite-host immunity and may yield important implications for choosing worm-derived antigens for the development of anti-schistosome vaccines.

Transforming growth factor-β, basement membrane components and heparan sulphate proteoglycans in experimental hepatic schistosomiasis mansoni

Abstract In an attempt to elucidate further the immunopathological pathways that underlie fibrogenesis induced by Schistosoma mansoni, we have studied the distribution of basement membrane compounds, heparan sulphate proteoglycans (HSPG) and the fibrogenic cytokine transforming growth factor (TGF)-β in two models of experimental schistosomiasis mansoni (experimental murine infection and synchronous granulomas induced by injection of egg-antigen-coupled beads into the caecal vein). Deposition of the basement membrane proteins type IV collagen, laminin and entactin in schistosomal granulomas was seen 3 days after the implantation of egg-antigen-coupled beads in the liver and persisted over time (32 days). Up-regulation of the membrane-bound HSPG syndecan-1 was observed in the schistosomal granuloma. These syndecan-1-immunoreactive cells represented a distinct subpopulation of granuloma cells; they were different from both mature, unstimulated B-cells (CD40-positive) and endothelial cells (CD105-positive). Deposition of the matrix HSPG perlecan within the granuloma was most prominent 8–16 days after injection. TGF-β expression was observed in acute (8 weeks) and chronically (13 weeks) infected mice, mainly at the periphery of the schistosomal granuloma and on Kupffer cells in the liver parenchyma. From these observations, we infer that schistosomal fibrosis is composed of various groups of matrix components and that TGF-β, which is secreted by granuloma cells, is one of the fibrogenic mediators in schistosomal fibrogenesis.

Schistosomal granuloma modulation. I. Schistosoma mansoni worm antigens CAA and CCA prime egg-antigen-induced hepatic granuloma formation

Abstract Adult Schistosoma mansoni worms can positively modulate soluble egg antigen (SEA)-induced granulomas formed around SEA-coupled beads implanted in the liver. In this study, our aim was to further unravel the immunopathological characteristics of S. mansoni-worm-derived antigens in vivo. (a) Adult worm antigen (AWA)-coupled Sepharose beads, implanted into the liver, induced granulomas, containing numerous eosinophilic granulocytes and elicited marked periparticular fibrosis (composed of interstitial matrix proteins and basement membrane components). (b) Quantitative morphological analysis demonstrated that in naive mice, AWA-induced hepatic granuloma formation peaked in volume 16 days after injection of the beads. An accelerated response against AWA-coupled particles (peak volume at 8 days) was observed in mice carrying a single-sex, male S. mansoni infection. (c) When the granuloma volume induced by SEA-coupled beads in unisexually S. mansoni infected mice was compared to granulomas induced by beads laden with both SEA and AWA in unsensitized mice, no significant differences in granuloma volume were seen, indicating the existence of in vivo egg/worm antigen cross-sensitization. (d) Naive mice, sensitized with the worm antigens circulating anodic antigen (CAA) or circulating cathodic antigen (CCA), mounted a strongly accelerated response towards SEA-coupled beads implanted in the liver. We infer that, in vivo, worm antigens cross-sensitize with egg antigens and have both granulomogenic and fibrogenic characteristics. The S. mansoni soluble worm antigens CCA and CAA prime hepatic egg-antigen-induced granuloma formation possibly through the presence of immunogenic carbohydrates. These mechanisms lead to an accelerated response against SEA.

Schistosomal granuloma modulation

Abstract Recurrent experimental evidence indicates that schistosomal egg granuloma formation – at least in the murine model – results from a host response generated against both egg- and worm-derived antigens. Further experiments aimed at identifying the existence in vivo of cross-sensitization between Schistosoma haematobium worms and S. mansoni-derived egg antigens were performed with respect to S. mansoni egg antigen-induced granuloma formation and fibrogenesis in the liver. Male OF1 mice bisexually infected with S. haematobium or S. mansoni were hepatically challenged (cecal vein injection) with S. mansoni SEA (soluble egg antigen)-coupled Sepharose beads at the end of prepatent infection (8–10 days prior to the start of egg deposition). The mean granuloma volume (MGV) of in-vivo-generated synchronized hepatic granulomas (8 days old) and the fibrotic response were estimated. Just like S. mansoni-infected rodents, mice carrying an S. haematobium infection generated an accelerated hepatic granulomogenesis [respective MGVs 4.72 ± 0.56 and 5.41 ± 0.75 × 106 μm3; P < 0.0001 versus unsensitized (MGV 3.00 ± 0.40 × 106 μm3) mice] and an enhanced fibrotic response against S. mansoni SEA. They also had significantly enlarged spleens (P < 0.0001) and moderately enlarged livers (P = 0.02) as compared with S. haematobium-infected mice that were not challenged with SEA. From these observations we infer that in vivo, S. haematobium worms can positively modulate S. mansoni egg antigen-induced granuloma formation and hepatic fibrogenesis, resulting in more severe liver pathology.

Adhesion molecules in intestinal Schistosoma mansoni infection

Abstract Adhesion molecules constitute essential elements in inflammation, mediating various cellular interactions. We investigated the expression of adhesion molecules mediating cell-cell [intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1)] and cell-matrix interactions [very late antigen-4 (VLA-4), VLA-6, and syndecan-1] in intestinal granulomas of mice infected with the parasite Schistosoma mansoni. Up-regulation of ICAM-1, LFA-1, and VLA-4 was seen in ileal and colonic granulomas, at both the acute (8 weeks postinfection) and the chronic stage (13–16 weeks postinfection). Up-regulation of VLA-6 was absent in all intestinal granulomas. Syndecan-1 immunoreactive (antigen-driven) B-lymphocytes were seen in the proximity of egg-antigen-laden macrophages in the inner part of ileal and colonic granulomas, although B-cells are considered to be absent in ileal granulomas. Estimation of intestinal granuloma volumes demonstrated the lack of down-modulation observed in ileal granulomas. From our results we infer that adhesion molecules constitute important elements in schistosomal intestinal granuloma formation. Organ-related differences between hepatic and intestinal granulomas exist (e.g., granuloma volume), but these differences are not morphologically reflected in a differential expression of the adhesion molecules ICAM-1, LFA-1, and VLA-4. Syndecan-1 immunoreactive B-lymphocytes also appear to be involved in ileal granuloma formation.

Expression of intercellular adhesion molecule-1 and lymphocytefunction-associated antigen-1 in experimental Schistosoma mansoni infection and in synchronous periparticular hepatic granulomas in mice: immunohistochemistry, confocal laser scanning microscopy, and immunoelectron microscopy

Abstract Adhesion molecules play an important role in inflammatory and immunological responses. We assessed the expression pattern of intercellular adhesion molecule-1 (ICAM-1) and lymphocytefunction-associated antigen-1 (LFA-1) in the livers of mice experimentally infected with Schistosoma mansoni and in synchronous hepatic granulomas induced by injection of soluble egg antigen (SEA)-coupled Sepharose beads in a mesenteric vein of mice. By immunohistochemistry, confocal laser scanning microscopy, and immunoelectron microscopy, ICAM-1 was localized on endothelial cells, sinusoidal-lining cells (Kupffer cells and sinusoidal endothelium), the hepatocyte cell membrane facing Disses space, and inflammatory cells in the granuloma. LFA-1 was visualized on the inflammatory cells of the granuloma and on phagocytic sinusoidal-lining cells, most likely Kupffer cells. ICAM-1- and LFA-1-immunoreactive cells were present in the granuloma as early as at 3 days after injection of SEA-coupled beads and persisted with time. As granulomas became older, nonimmunoreactive granuloma cells appeared. We conclude that adhesion molecules play an important role in the genesis of the schistosomal granuloma.

Confessed abusive blunt head trauma.

AbstractIt is generally accepted that terms referring to specific craniocerebral injury mechanisms must be replaced by the more general term abusive head trauma (AHT). Although blunt impact trauma remains an essential part of AHT, it has received far less attention in the literature than shaken-impact injuries. The current article presents 19 confessed cases of a series of 47 highly suspected AHT cases. Of these, 13 were confessed shaken-impact cases, and the other 6 confessed blunt trauma cases. There were no significant differences in the appearance of subdural hematoma, which was present in each case. Retinal hemorrhage, which was present in 10 of the 13 shaken-impact cases in which an ophthalmologic examination was conducted, occurred in 2 of the 6 blunt trauma cases. In 1 case, retinal hemorrhage probably had of metabolic origin. Skull fractures with an overlying subgaleal hematoma and a subdural hematoma below the fracture side were found in 5 of the blunt trauma cases but was also seen in the 2 shaken-impact cases with a skull fracture. The most important finding was a lucid interval (LI) in 3 blunt AHT cases. An LI does not seem to occur in shaking injuries because of the immediate and persistent effect of brain damage that such injuries involve. Therefore, LI makes it important to conduct a detailed investigation of the clinical course in time in suspected AHT cases.

Can post-mortem CT reliably distinguish between drowning and non-drowning asphyxiation?

PurposeThe aim of this study is to evaluate whether previously reported post-mortem CT findings in drowning can reliably distinguish drowning from asphyxiation by any other manner.Materials and methodsCases (n = 14) were corpses with cause of death determined as drowning by concordant autopsy findings and physical and circumstantial evidence. Controls (n = 11) were corpses in which the cause of death was defined as asphyxiation by any other manner than submersion in a liquid. Images were evaluated for the presence of fluid in paranasal sinuses, mastoid air cells and lower airways, frothy foam in the upper airways, ground-glass opacity of the lung parenchyma, the height of the right hemi-diaphragm, the interpulmonary distance at the level of the aortic valve, the mean density of intracardiac blood, and gastric and esophageal contents. Descriptive statistics, Fisher’s exact test, and Student’s t test were used when appropriate.ResultsOnly the height of the right hemi-diaphragm differed significantly (p = 0.045) between cases (mean 5.4) and controls (mean 4.3). Other findings were not significantly different between both groups.ConclusionOur results indicate that it is not possible to reliably distinguish drowning from non-drowning asphyxiation on CT, because many findings in drowning were also present in non-drowning asphyxiation. CT indicators for drowning as the cause of death should therefore be defined with great caution, keeping in mind that they are not specific to only a single cause of death.