Complement activation contributes to ventilator-induced lung injury in rats.

The complement system contributes to ventilator induced lung injury (VILI). We hypothesized that pretreatment with the C1 esterase inhibitor (C1INH) Berinert® constrains complement activation consecutively inducing improvements in arterial oxygenation and histological pulmonary damage. At baseline, male Sprague-Dawley rats underwent mechanical ventilation in a conventional mode (PIP 13 cm H2O, PEEP 3 cm H2O). In the Control group, the ventilator setting was maintained (Control, n = 15). The other animals randomly received intravenous pretreatment with either 100 units/kg of the C1-INH Berinert® (VILI-C1INH group, n = 15) or 1 ml saline solution (VILI-C group, n = 15). VILI was induced by invasive ventilation (PIP 35 cm H2O, PEEP 0 cm H2O). After two hours of mechanical ventilation, the complement component C3a remained low in the Control group (258 ± 82 ng/ml) but increased in both VILI groups (VILI-C: 1017 ± 283 ng/ml; VILIC1INH: 817 ± 293 ng/ml; P < 0.05 for both VILI groups versus Control). VILI caused a profound deterioration of arterial oxygen tension (VILI-C: 193 ± 167 mmHg; VILI/C1-INH: 154 ± 115 mmHg), whereas arterial oxygen tension remained unaltered in the Control group (569 ± 26 mmHg; P < 0.05 versus both VILI groups). Histological investigation revealed prominent overdistension and interstitial edema in both VILI groups compared to the Control group. C3a plasma level in the VILI group were inversely correlated with arterial oxygen tension (R = -0.734; P < 0.001). We conclude that in our animal model of VILI the complement system was activated in parallel with the impairment in arterial oxygenation and that pretreatment with 100 units/kg Berinert® did neither prevent systemic complement activation nor lung injury.

SNAP-25 is abundantly expressed in enteric neuronal networks and upregulated by the neurotrophic factor GDNF.

Control of intestinal motility requires an intact enteric neurotransmission. Synaptosomal-associated protein 25 (SNAP-25) is an essential component of the synaptic vesicle fusion machinery. The aim of the study was to investigate the localization and expression of SNAP-25 in the human intestine and cultured enteric neurons and to assess its regulation by the neurotrophic factor glial cell line-derived neurotrophic factor (GDNF). SNAP-25 expression and distribution were analyzed in GDNF-stimulated enteric nerve cell cultures, and synaptic vesicles were evaluated by scanning and transmission electron microscopy. Human colonic specimens were processed for site-specific SNAP-25 gene expression analysis and SNAP-25 immunohistochemistry including dual-labeling with the pan-neuronal marker PGP 9.5. Additionally, gene expression levels and distributional patterns of SNAP-25 were analyzed in colonic specimens of patients with diverticular disease (DD). GDNF-treated enteric nerve cell cultures showed abundant expression of SNAP-25 and exhibited granular staining corresponding to synaptic vesicles. SNAP-25 gene expression was detected in all colonic layers and isolated myenteric ganglia. SNAP-25 co-localized with PGP 9.5 in submucosal and myenteric ganglia and intramuscular nerve fibers. In patients with DD, both SNAP-25 mRNA expression and immunoreactive profiles were decreased compared to controls. GDNF-induced growth and differentiation of cultured enteric neurons is paralleled by increased expression of SNAP-25 and formation of synaptic vesicles reflecting enhanced synaptogenesis. The expression of SNAP-25 within the human enteric nervous system and its downregulation in DD suggest an essential role in enteric neurotransmission and render SNAP-25 as a marker for impaired synaptic plasticity in enteric neuropathies underlying intestinal motility disorders.

GDNF induces synaptic vesicle markers in enteric neurons.

Regulation of intestinal motility depends on an intact synaptic vesicle apparatus. Thus, we investigated the expression of the synaptic vesicle markers synaptophysin and synaptobrevin in the human enteric nervous system (ENS) and their regulation by glial cell line-derived neurotrophic factor (GDNF) in cultured enteric neurons. Full-thickness specimens of the human colon were assessed for expression of synaptophysin and synaptobrevin and neuronal localization was assessed by dual-label immunocytochemistry with PGP 9.5. Effects of GDNF on both synaptic markers were monitored in enteric nerve cell cultures and the presence of varicosities was determined by applying electron microscopy to the cultures. Human colonic specimens showed immunoreactivity for synaptophysin and synaptobrevin in both myenteric and submucosal ganglia as well as in nerve fibers. Both synaptic vesicle markers co-localized with the neuronal marker PGP 9.5 and exhibited granular accumulation patterns in the human and rat ENS. In cultured rat myenteric neurons GDNF treatment promoted expression of both synaptic vesicle markers and the formation of neuronal varicosities. The regulation of synaptophysin and synaptobrevin in enteric neurons by GDNF argues for the induction of functional neuronal networks in culture characterized by an increase of synaptogenesis.

[Inflammatory pseudotumours of the liver associated with Crohn's disease: a possible pitfall in contrast-enhanced ultrasound]. / Inflammatorische Pseudotumoren der Leber bei Morbus Crohn: ein möglicher Pitfall bei der Echokontrast-Sonografie.

Pseudotumours of the liver are rare entities, which cannot be differentiated from malignant liver lesions by contrast-enhanced ultrasound (CEUS) or radiological imaging. We report of a 22-year-old man with persistant abdominal pain and fever after appendectomy. Fundamental ultrasound showed an inflammatory swelling of the terminal ileum, suspicious of Crohn's disease as well as two solid liver lesions and left portal vein thrombosis. The enhancement patterns of both liver tumors on CEUS showed inhomogeneous isoenhancement during the arterial and portal phases and hypoenhancement during the late phase, mimiking malignant lesions. Ultrasound-guided biopsy, histology and follow-up, however, confirmed the diagnosis of hepatic inflammatory pseudotumour (IPT). When diagnosing a pseudotumour of the liver, the rare coincidence with chronic inflammatory bowel disease should be considered.

[Treatment of fibrous dysplasia]. / Therapiekonzepte bei fibröser Dysplasie.

BACKGROUND: The fibrous dysplasia is a rare disease of bone metabolism. Most common on the extremities, its appearance at the skull base is rare but of importance in clinical otorhinolaryngology. The patients suffer from problems such as cosmetical limitation, recurrent pain and in later phases from functional losses which result from bone dysplasia. METHODS: Based on the experiences of these cases and the study of the current literature we would like to suggest a stage dependent therapy concept that includes conservativ options as well as surgical methods which are divided into curative and palliative surgical treatments. PATIENTS: The Department of Otorhinolaryngology in Bochum has been involved in the therapy of eight patients suffering from fibrous dysplasia which have been treated and examined from six to twenty years. RESULTS: Treatment in fibrous dysplasia can be efficient in increasing quality of life. A complete healing is rare and not the main target of treatment. CONCLUSIONS: The staged therapy of fibrous dysplasia consists of conservative and surgical treatment and the combination of both. The decision for an individual therapy concept might be difficult and often requires close interdisciplinary cooperation.

[Frequency and morphology of tuberculosis in autopsies: increase of active forms]. / Häufigkeit und Morphologie der Tuberkulose bei Obduktionen: Zunahme aktiver Formen.

BACKGROUND AND OBJECTIVE: Tuberculosis is still a relevant infectious disease, which is clinically often not diagnosed during a patient's lifetime. We investigated the frequency of tuberculosis in autopsies. METHODS: 3947 autopsy reports from the Institute of Pathology at the Ruhr University Bochum,were analysed for the period from 1990 to 2004. Tuberculosis was mentioned in 148 reports. RESULTS: 55 (1.39%) cases showed relevant tuberculosis. In 39 (70.9%) cases further other relevant diseases were diagnosed. Lung involvement was grossly present in 52 (94.6%) cases. Of the active forms, 16 (29.1%) showed acinar nodal foci, 15 (27.3%) so-called early cavities and 10 (18.2%) caseous pneumonia. Miliary tuberculosis was found in 8 (14.5%) cases, tubercular meningoencephalitis in 7 (12.7%), and tuberculosis of the spine in 4 (7.3%). The inactive forms showed scarring or pleural adhesions (n = 30, 54.5%), late cavities and mycetomas (n = 9, 16.3%). Active forms of tuberculosis were more frequent in the age groups from 30 to 59 and 80 to 99 years. In the last period of the study an increase in active forms (22:2 = 91.7% vs. 18:13 = 58.1%, p = 0.0065) was noted, while 10 of 22 (45.5%) cases were not detected clinically. In 13 (0.33%) cases tuberculosis had been diagnosed as requiring treatment but no tuberculosis was found at autopsy. CONCLUSION: Early diagnosis of tuberculosis is very important for patients and for inhibiting a possible spread of bacteria, especially considering the increase in frequency of active forms of tuberculosis. Autopsies are still indispensable for providing quality control and disease statistics.

[The duty of notification for pathologists according to the infectious disease control act. Tuberculosis as dominating disease]. / Meldepflicht des Pathologen nach dem Infektionsschutzgesetz : Tuberkulose als dominierende Erkrankung.

The Infectious Disease Control Act enacted in Germany in 1.1.2001 led to a duty of notification also for institutes of pathologic-anatomical diagnostics. All reports within 45 months after enacting concerning diseases and agents being subject to registration were evaluated. Among the notifiable diseases with fatal outcome ( section sign 6) belonged 3 cases of Meningococcus sepsis, 13 of tuberculosis und 5 cases of Creutzfeldt-Jacob disease. During lifetime 54% of tuberculosis cases remained undetected. Notifiable agents ( section sign 7.1) concerned 92 times Mycobacterium-tuberculosis-complex, twice Influenza Virus and one case of Cryptosporidiosis and Giardia lamblia each. Six Echinococcus granulosus cysts were reported ( section sign 7.2). Notification needs exact diagnosis of infectious diseases and agents being subject of registration. By this pathologists participate in the control of infectious diseases.

Hypoxia-induced intrauterine growth retardation: effects on pulmonary development and surfactant protein transcription.

BACKGROUND AND OBJECTIVES: Preterm infants with intrauterine growth retardation (IUGR) reveal an increased risk for the development of acute and chronic pulmonary disorders, i.e. bronchopulmonary dysplasia (BPD). In order to investigate the effect of IUGR on pulmonary development, an easily reproducible animal model for fetal growth restriction has been established using hypoxia as a sole intervention in the last third of pregnancy. METHODS: Date-mated mice were randomly assigned to either being kept at a fraction of inspired oxygen (FiO2) of 0.10 (hypoxic group) starting at day 14 or under normoxic conditions until day 17.5 of gestation (control group). Variables of somatic growth were assessed and standardized histomorphometric analyses of pulmonary tissue were performed. Expression of surfactant proteins (SP)-A, -B, -C and -D was determined by quantitative rt-PCR as biochemical indicators for lung development and maturation. RESULTS: Fetuses were delivered preterm at 0.87 of gestation. Those grown under hypoxic conditions revealed significantly lower birth weights (median: 0.69 vs. 0.97 g in controls; p < 0.001), body lengths (median: 17.5 vs. 20.2 mm in controls; p < 0.001) and fronto-occipital diameters (median: 9.4 vs. 10.1 mm in controls; p < 0.001) compared to controls. Histomorphometric analyses were found to be without significant differences between both groups. On the transcriptional level, however, mRNA expression of SP-A, -B and -C but not SP-D could be shown to be significantly reduced in hypoxic fetuses compared to normoxic controls. CONCLUSIONS: In conclusion, hypoxic conditions from day 14 to 17.5 led to IUGR in preterm mice and to significant alterations of the developing surfactant system. We speculate restricted development of SP gene expression to be a causal factor for the increased risk of acute and chronic pulmonary disorders in preterm infants with IUGR.

Ultrastructural changes in the cilia of experimental mice.

Mice lacking dynein arms in the cilia were examined; the strain was obtained by inactivation of dynein heavy chain gene in chromosome 7. The cilia of these mice were examined by electron microscopy and compared to the cilia of random-bred mice. No statistically significant differences or typical disorders in the outer or inner dynein arms were detected. The number of inner dynein arms was lower, in some cilia secondary changes presenting as swelling of the outer part of the ciliary membrane or formation of complex cilia were seen.

[Reconstruction of bile duct lesions by an autologous vein graft and a bio-degradable endoluminal stent in an animal model: technique and clinical impact]. / Rekonstruktion von Gallengangsdefekten mit autologem Veneninterponat und resorbierbarem Stent im Tiermodell: Technik und klinische Bedeutung.

INTRODUCTION: In this study a new treatment of bile duct lesions was investigated. A segment of the bile duct was replaced by an autologous venous interponate which had been endoluminally stented with a braided bio-degradable stent. METHODS: A total of 18 pigs (20-28 kg) was divided into three equal groups (I-III). In each group a 2 cm segment of the jugular vein was harvested. The animals in Group I (vein group, n = 6) underwent resection of a 2 cm long segment of the common bile duct which was replaced solely by the venous interponate, in Group II (stent group, n = 6) the venous interponate had been endoluminally stented by a braided bio-degradable stent. Group III (control group, n = 6) underwent only a circular mobilization of the common bile duct. Postoperatively survival rate, general condition as well as the weight were observed and checked for 6 months. During surgery and finally after sacrifice after 6 months blood and tissue samples were taken and semiquantitatively scored concerning grade of inflammation and fibrosis. RESULTS: In the stent and control group all animals survived in good condition. 3 pigs of the vein group died within 3 weeks showing signs of biliary peritonitis, another one died due to a high grade stenosis of the common bile duct with secondary biliary cirrhosis after 4 months. In the stent group all animals survived until sacrifice after 6 months. On examination the venous interponate was laminated with bile duct epithelium showing the diameter of the implanted stent. CONCLUSION: The reconstruction of bile duct lesions by a venous interponate in combination with a bio-degradable stent is easy to perform and represents a clinically interesting alternative to the biliodigestive anastomosis because of the preservation of the sphincter oddi. After 6 months the stent is completely absorbed and the venous interponate is laminated with bile duct epithelium.

Detection of Chlamydophila pneumoniae in mouse respiratory ciliated epithelium using targeted sections of the lung tissue.

Chlamiophila pneumoniae were detected in targeted sections of mouse lung tissue by means of transmission electron microscopy and immunofluorescent staining. Incorporation of microorganisms into the axonemal matrix of cilia was observed 24 h after infection. The ciliary axoneme was characterized by pronounced swelling. At the late stages Chlamiophila pneumoniae were present in cytoplasmic vacuoles. Structural abnormalities and dysfunction of mucociliary clearance followed by incorporation of Chlamiophila pneumoniae into the cytoplasm of epitheliocytes were revealed in the early stage of infection. The proposed method allows studying the very early events of Chlamiophila pneumoniae infection.

Infection of murine precision cut lung slices (PCLS) with respiratory syncytial virus (RSV) and chlamydophila pneumoniae using the Krumdieck technique.

The Krumdieck technique allows the investigation of the so-called precision cut lung slices (PCLS) with a special microtome. It is thus possible to evaluate morphologic changes over a longer period of time using only a small group of animals. Chlamydophila pneumoniae (Cp) and respiratory syncytial virus (RSV) proved to be important causes of pneumonia, rhinitis and exacerbations of asthma bronchiale, as well as of lower respiratory tract infections in young children. PCLS should be tested for their suitability as an in vitro model for these infections. The PCLS were infected with Cp and RSV over different periods of time. Investigations were carried out by light and transmission electron microscopy (TEM). Furthermore, immunofluorescence (IF) studies with antibodies against bacterial or viral proteins and cell-specific markers were done using confocal laser scanning microscopy (CLSM). Non-infected and infected PCLS showed a well-preserved morphology up to 72 hours. After short infection intervals, typical inclusions of Cp or RSV were detected in vacuoles of different cell types. Infection and cell types could be verified using IF. Cytopathic effects were not prominent. Ciliary beat was detectable up to 96 hours after infection. This in vitro technique offers the possibility of studying mechanisms and effects of bacterial and viral infections on viable tissue complexes.

Pulmonary administration of perfluorodecaline- gentamicin and perfluorodecaline- vancomycin emulsions.

The aim of this study was to examine pharmacokinetics and pulmonary antibiotic tissue concentrations (PATC) of gentamicin and vancomycin after intrapulmonary administration of a perfluorodecaline (PFD)-gentamicin and a PFD-vancomycin emulsion during partial liquid ventilation (PLV). PLV was initiated in 19 healthy rabbits and 18 surfactant-depleted rabbits. The animals were randomized to receive either 5 mg/kg gentamicin and 15 mg/kg vancomycin intravenously, or 5 mg/kg gentamicin intrapulmonary, or 15 mg/kg vancomycin intrapulmonary. Antibiotic plasma levels were measured after 15, 30, 45, and 60 min, and hourly thereafter. After 5 h animals were sacrificed and lungs were removed to evaluate PATC and histology. PATC were significantly higher after intrapulmonary administration of both gentamicin and vancomycin. In healthy rabbits, peak plasma concentrations were lower and 5 h plasma concentrations were higher after intrapulmonary administration, whereas plasma concentrations were not different in surfactant-depleted rabbits. There were no differences in lung histology, hemodynamics, lung mechanics, or gas exchange between the treatment groups. We conclude that during PLV, higher PATC can be achieved after intrapulmonary administration of PFD-antibiotic emulsions compared with intravenous administration of the same dose without apparent short-term adverse effects. We speculate that intrapulmonary antibiotic administration during PLV may be beneficial in treating severe pneumonia.

Transformation of Hodgkin's disease to high-grade B-cell lymphoma: remission after Rituximab monotherapy.

PURPOSE: We demonstrate the usefulness of immunotherapy with the CD20 antibody Rituximab in a case of transformation of Hodgkin's disease (HD) to high-grade non-Hodgkin's lymphoma (NHL). CASE REPORT: A 45-year-old women suffering from lymphocyte predominant HD of paragranuloma type (stage IVb) since 1995 showed mediastinal relapse despite of 6 cycles of chemotherapy following the COPP/ABVD-protocol in 1998. Again complete remission could be achieved after escalated BEA-COPP II therapy in May 1998. Six months later chest radiograph and CT depicted pulmonary nodules. The non-typical resection of the lung revealed pulmonary involvement of a high-grade T-cell rich large B-cell lymphoma with 100% of the tumoral cells CD20 positive. Since the symptoms occurred shortly after the BEA-COPP-escalated protocol chemotherapy resistance had to be assumed. Because of this problems and supported by the refusal of a high-dose chemotherapy with stem-cell transplantation by the patient we decided to perform a mono-immunotherapy with the monoclonal CD20 antibody Rituximab. Today, 14 months later, the patient is still in complete remission including the absence of B symptoms. CONCLUSIONS: Immunotherapy against CD20 positive cells in cases of sequential HD and NHL seems to be an effective therapy in chemotherapy resistant cases because of the suspected clonally relation of both diseases.