Effect of decorin on overcoming the extracellular matrix barrier for oncolytic virotherapy.

The pressing challenge for contemporary gene therapy is to deliver enough therapeutic genes to enough cancer cells in vivo. With the aim of improving viral distribution and tumor penetration, we explored the use of decorin to enhance viral spreading and tumor tissue penetration. We generated decorin-expressing replication-incompetent (dl-LacZ-DCNG, dl-LacZ-DCNQ and dl-LacZ-DCNK) and replication-competent (Ad-DeltaE1B-DCNG, Ad-DeltaE1B-DCNQ and Ad-DeltaE1B-DCNK) adenoviruses (Ads). Point mutants of decorin gene (DCNG), DCNK and DCNQ, have a negative and moderate binding affinity to type-I collagen fibril, respectively. In both tumor spheroids and established solid tumors in vivo, tissue penetration potency of dl-LacZ-DCNG was greatly enhanced than those of dl-LacZ, dl-LacZ-DCNQ and dl-LacZ-DCNK, and this enhanced tissue penetration effect derived from decorin-expressing Ad was dependent on the binding affinity of decorin to collagen fibril. Expression of DCNG enhanced viral spread of replicating Ad, leading to improved tumor reduction and survival benefit. Moreover, the tumoricidal effects of Ad-DeltaE1B-DCNQ and Ad-DeltaE1B-DCNK were lessened, as the binding affinity to collagen was decreased, showing that the increased cancer cell cytotoxicity was driven by the action of decorin on extracellular matrix (ECM). Furthermore, Ad-DeltaE1B-DCNG substantially decreased ECM components within the tumor tissue. Finally, intratumoral injection of Ad-DeltaE1B-DCNG in primary tumor site greatly reduced the formation of B16BL6 melanoma cell pulmonary metastases in mice. Taken together, these data show the utility of decorin as a dispersion agent and highlight its utility and potential in improving the efficacy of replicating Ad-mediated cancer gene therapy.

Bridging the gap: heparan sulfate and Scube2 assemble Sonic hedgehog release complexes at the surface of producing cells.

Decision making in cellular ensembles requires the dynamic release of signaling molecules from the producing cells into the extracellular compartment. One important example of molecules that require regulated release in order to signal over several cell diameters is the Hedgehog (Hh) family, because all Hhs are synthesized as dual-lipidated proteins that firmly tether to the outer membrane leaflet of the cell that produces them. Factors for the release of the vertebrate Hh family member Sonic Hedgehog (Shh) include cell-surface sheddases that remove the lipidated terminal peptides, as well as the soluble glycoprotein Scube2 that cell-nonautonomously enhances this process. This raises the question of how soluble Scube2 is recruited to cell-bound Shh substrates to regulate their turnover. We hypothesized that heparan sulfate (HS) proteoglycans (HSPGs) on the producing cell surface may play this role. In this work, we confirm that HSPGs enrich Scube2 at the surface of Shh-producing cells and that Scube2-regulated proteolytic Shh processing and release depends on specific HS. This finding indicates that HSPGs act as cell-surface assembly and storage platforms for Shh substrates and for protein factors required for their release, making HSPGs critical decision makers for Scube2-dependent Shh signaling from the surface of producing cells.

Heroin and opiate emergencies in Vienna: analysis at the municipal ambulance service.

Vienna suffered an epidemic of heroin abuse in recent years, with drug-deaths due to opioids increasing from 62 in 1991 to 143 in 1993. The aim of this study was to make observations about illicit opioid-use with the ambulance service as a data source. From June 1994 to August 1995, the structured run records of the ambulance service were reviewed. Those with a presumptive diagnosis of "heroin or opiate" overdose were collected, characteristics of emergencies and patients were analyzed. The run records demonstrated a large number of non-fatal emergencies due to opioids, involving 528 men and 179 women in 1087 emergencies. These emergencies were on the average 6.8 times as prevalent as drug-fatalities. A group of 189 persons could be identified, who caused 52.2% of all emergencies and showed a threefold mortality rate during the observation period. In Vienna, the records of the municipal ambulance service provided valuable insights on opioid-abuse. We suggest local analysis of non-fatal emergencies due to opioids, as this might lead to a new source of information on illicit abuse of these drugs.

After antagonization of acute opiate overdose: a survey at hospitals in Vienna.

This study describes the clinical management and characteristics of people who, following acute opioid overdose, are taken to hospital after efficient antagonization by the pre-hospital emergency service. In addition, it defines areas of interest for further research. Over a 4-month period (September-December 1993) we collected data by a structured protocol sheet on patients' characteristics, anamnestic data on abuse and emergencies, clinical presentation, treatment by specific antidote and routine laboratory investigations. Outcome was verified by retrospective review of prehospital and forensic data. We studied 77 subjects, predominantly young males, who were involved in 83 emergencies, mostly occurring at weekends. In more than 60% of cases a single administration of specific antidote sufficed to stabilize the patients; 64% of patients left hospital against medical advice after an average stay of less than 6 hours; 46% denied daily opioid abuse and half the subjects, especially younger drug-users, seemed interested in counselling. This hospital-based study did not provide reliable data on the epidemiology of opioid overdose. Clinical management is determined by experience, pragmatism and beliefs. Efforts towards secondary prevention of drug problems at emergency departments might be warranted, and further research on pattern and management of opioid overdose is needed.

Efficacy of different antihypertensive drugs in the emergency department.

Hypertensive crises are a commonly observed problem in an emergency department. The aim of the study was to evaluate the efficacy and safety of different antihypertensive agents in the treatment of patients with hypertensive crises. 168 patients (mean age: 52 +/- 12 years) admitted to the emergency department with a hypertensive urgency (systolic (SBP) blood pressure > 210 mm Hg and/or diastolic (DBP) blood pressure > 110 mm Hg) or a hypertensive emergency (DBP > 100 mm Hg and evidence of end-organ damage) were included into the study protocol. Blood pressure (BP) was measured every 5 min automatically using a noninvasive BP measurement unit. After a resting period of 30 min the patients received the following drugs: 5 mg enalaprilat intravenous (n = 43) or 25 mg urapidil intravenous (n = 48) or 10 mg nifedipine-capsule sublingual (n = 47) or 2 x 5 mg nifedipine-spray sublingual (n = 30). The aim of treatment was to reduce SBP below 180 mm Hg and DBP below 95 mm Hg within 45 min after start of treatment. When evaluating the response rates the highest rate was observed in the urapidil group (96%). The response rate of enalaprilat and both preparations of nifedipine were similar (70-72%). The rate of major side effects was higher in the urapidil compared to the other drugs (4% vs 2% in the nifedipine-group or 0% in the enalaprilat-group). All four drugs are suitable in the treatment of patients with hypertensive crisis in the emergency department. Urapidil should be used as a first choice drug in critically ill patients with hypertensive crisis due to its higher response rate.

[Evaluation system in assessing radiological examinations of clinical significance for the patient]. / Bewertungssystem zur Erfassung des Stellenwertes radiologischer Leistungen für die klinische Arbeit am Patienten.

PURPOSE: The purpose of this study was to test a scoring system and its utility for the assessment of the clinical significance of radiological examinations. MATERIAL AND METHODS: The reports of all diagnostic studies obtained in patients with a clinical suspicion of pancreatitis, urinary tract obstruction, biliary or gallbladder diseases, ileus and gastrointestinal perforation were scored prospectively in 4 categories from 0 to 3. These categories were related to the ability to establish a definite diagnosis, to exclude suspected disease, to influence further diagnostic work-up and therapeutic decision-making. RESULTS: This scoring system showed significant differences of the efficacy of different imaging modalities in these 5 clinical settings. CONCLUSION: This scoring system was helpful to determine the clinical significance of radiological examinations.

Referral of outpatients with non-traumatic conditions for radiographic examinations in an emergency department.

The aim of the study was to evaluate the referral of outpatients with non-traumatic conditions for radiographic examinations and to assess the impact of the radiological report on the patient's management in an emergency department. In a prospectively designed study, 1223 X-ray examinations of 1116 non-trauma outpatients (640 males, 476 females; mean age: 44 +/- 18 years) requested over a 10-month period were evaluated. Patients were classified into four groups according to the presenting complaints (respiratory, abdominal, neurological or non specific symptoms). Analysis of data included the influence of age and presenting symptoms on the likelihood of abnormal radiological findings and the impact of the radiological result on the further management of the patient. In 455 (40.8%) patients an abnormal radiological result was observed. The likelihood of an abnormal radiological findings increased with age (age < 40 years: 33%; age > 40 years; 47%; p < 0.05). Whereas the rate of abnormal radiological findings was high in patients with specific symptoms (respiratory: 69%; abdominal: 37%; neurological: 38%), in patients with non-specific symptoms only 25% of all radiological examinations revealed an abnormality. The radiological result had an impact on the further management in 948 (85%) patients. As 45% of all radiographic examinations revealing a normal radiological result had a clinical impact, normal radiological reports are just as helpful as abnormal radiological findings in the management of non-trauma outpatients in an emergency department. Thus, we assume that the radiological result has a major impact on the management of non-trauma outpatients in the emergency department.

[Carbamazepine poisoning: protracted course with development of intestinal atony and hepatic toxicity]. / Intoxikation mit Carbamazepin: Protrahierter Verlauf unter Entwicklung von Darmatonie und Hepatotoxizität.

A 43 year-old female patient with a history of manic-depressive illness and prophylactic carbamazepine (CBZ) medication ingested a potentially lethal overdose of 20 g of the substance. Neurotoxic symptoms reached full intensity after about 24 hours. Subsequently, the patient developed a gastrointestinal atony, which proved to be refractory to treatment for several days. Moreover, there was an increase of bilirubin. Parallel to this we observed the persistence-and even intermittent re-increase--of toxic serum CBZ concentrations for one week with corresponding protracted clinical symptomatology.

Effect of low dose sedation with diazepam on ventilatory response at moderate altitude.

UNLABELLED: Benzodiazepines are in wide use in out of hospital emergency medicine in at sea level, as well as in alpine rescue services. Ventilatory depression is a common side effect of high dose intravenous benzodiazepines. The objective of our study was to evaluate the effect of low dose benzodiazepines on ventilation at moderate altitude. DESIGN: Randomized, double blind, placebo controlled crossover trial. Blood gas analysis of 8 healthy subjects was performed before and one hour after 5 mg of oral diazepam or placebo at 171 m and at 3,000 m altitude. PaO2 and PaCO2 before and after diazepam or placebo medication at 171 m did not change significantly. At 3,000 m altitude. PaO2 decreased and PaCO2 increased significantly one hour after 5 mg of oral diazepam (PaO2 64.7 +/- 3.4 vs. 60.0 +/- 1.5 mmHg, PaCO2 28.3 +/- 2.3 vs. 33.3 +/- 1.8 mmHg, p < 0.05). Placebo did not influence PaO2 and PaCO2. CONCLUSION: Low dose diazepam inhibits ventilatory adaptation to mild hypoxia at moderate altitude. Caution in benzodiazepine use in moderate altitude is therefore necessary.

[Local complications after poisonous snake bite]. / Lokale Komplikationen nach einem Giftschlangenbiss.

The case of a zoo keeper who was bitten on the left finger by a venomous snake (Vipera xanthina) is reported. The administration of antivenom prevented the development of systemic poisoning but had no effect on the extent of the local complications. A compartment syndrome with a concomitant severe reaction at the bite site required fasciotomy of the upper and lower arm. The extensor tendon of the involved finger ruptured spontaneously, many weeks after wound healing was completed. Therefore, delayed local complications following snake bites may occur, even if signs of systemic poisoning are missing.

TGF-beta1 generates a specific multicomponent extracellular matrix in human coronary SMC.

BACKGROUND: Transforming growth factor (TGF-beta(1)) is postulated to play an important role in maintaining the structure and function of arterial tissue and protection against development of arteriosclerosis. The TGF-beta(1)-induced production of a stable extra-cellular matrix-rich plaque phenotype is suggested to be part of the protection against a switch to an unstable rupture-prone arteriosclerotic plaque. MATERIALS AND METHODS: This study addresses the question of whether the expression profile and the type of extra-cellular matrix (ECM) generated by TGF-beta(1) stimulation have the structural feature of a fibril-rich stable matrix. Seventeen genes codings for ECM components of human coronary smooth muscle cells (SMCs) after a 24-h stimulation by TGF-beta(1) have been analyzed. RESULTS: Real-time RT-PCR was used to quantify the mRNA of genes under investigation. It was found that after TGF-beta(1) stimulation (a) the up-regulation of COL1A1-specific mRNA was associated with increased [(3)H]proline incorporation into the alpha-1 and -2 chains of collagen type I, (b) the up-regulation of biglycan- and syndecan-1-specific mRNA corresponded to an increased [(35)S]sulphate and [4,5-(3)H]leucine incorporation into the biglycan molecule and to an increase of syndecan-1 protein, (c) the up-regulated FGF-2 gene accounted predominantly for the ECM-bound subfraction of FGF-2-protein and (d) fibronectin and thrombospondin exhibited a significantly higher mRNA level. In contrast collagen XIV, a minor collagen type, and the proteoglycan decorin were down-regulated. The down-regulated decorin changed its structure by elongation and reduced GlcA to IdoA epimerization of the dermatan sulphate side-chain as judged by [(35)S]sulphate metabolic labelling experiments. No significant changes in response to TGF-beta(1) were observed for the collagen types III, VI and XVI, for versican, perlecan and the syndecans-2 and -4. CONCLUSIONS: It was concluded from the data that the TGF-beta(1)-induced formation of a highly specific multicomponent extra-cellular matrix on coronary arterial SMCs could provide in vivo mechanical strength to the neointima in arteriosclerotic lesions and to the fibrous cap overlying the lipid core.

[Neuraminidase neutralizing antibodies in pigs with chronic Erysipelothrix rhusiopathiae infection (author's transl)]. / Uber das Vorkommen Neuraminidase-neutralisierender Antiköper bei chronisch rotlaufkranken Schweinen

Investigating sera from pigs with chronic E. rhusiopathiae infection relative gamma-globulin values, agglutination titers, and neuraminidase-neutralizing humoral antibodies were determined at the 12th and 15th, 11th and 14th, and 3rd and 6th month after infection. We found increasing gammaglobulin values and increasing neuraminidase neutralizing antibodies but decreasing agglutination titers. Few animals don't have antibodies against neuraminidase and died during the observation period. Furthermore immunodiffusion test demonstrated precipitating lines of antibodies against living E. rhusiopathiae cells and active neuraminidase. No or very weak bands were observed against heat killed cells and heat-inactivated enzyme.

Expression of fibroblast growth factors 1 and 2 in a human retinal pigment epithelium cell line (K1034).

The primary source of fibroblast growth factors (FGFs) is the retinal pigment epithelium (RPE). Investigations on FGF secretion in RPE primary cultures are hampered by the rapid run-down of cell vitality after a few passages. Therefore, long-term experiments require an alternative to primary cultures. We detected FGF-1 and FGF-2 in the established human K1034 cell line by immunohistochemistry. In addition, mRNA for both FGFs was found by RT-PCR. By immunohistochemistry, the signal was more pronounced with FGF-2 than with FGF-1. K1034 is capable of expressing both FGF-1 and FGF-2. With respect to these features, this cell line can be used as an alternative to primary cultured human RPE cells.

Management of lead foreign body ingestion.

Previous reports of lead foreign body (PbFB) ingestion by children include two cases of lead intoxication and two cases that resulted in death. It is generally accepted, however, that PbFB ingestion does not pose a risk of lead toxicity, provided that the PbFB is not retained in the gastrointestinal (GI) tract. Recent experience with two cases substantiates this belief. On separate occasions, two 9-year old girls ingested a PbFB. The first patient developed only mild, vague GI complaints. The highest reported blood lead level (BLL) was 15 mcg/dl. The second patient did not develop symptoms, and the highest reported BLL was 22 mcg/dl. Both patients passed the PbFB in the stool within two weeks. These cases suggest that PbFB ingestions can be treated in a manner similar to other uncomplicated foreign body ingestions.

Pulmonary embolism. II. Diagnosis and treatment.

Pulmonary embolism is an often underestimated, underdiagnosed, and undertreated disease. As symptoms and signs of pulmonary embolism are nonspecific, the diagnosis still remains a challenge to the attending physician. Diagnostic and therapeutic procedures depend on the clinical presentation of the patient. First we must suspect pulmonary embolism and consider its likelihood in the presence of a number of clinical signs and symptoms. Once pulmonary embolism is suspected, heparin should be administered. Additional basic support is mandatory if required. If the patient's hemodynamic situation is stable, available preferable noninvasive diagnostic options should be considered to confirm or rule out the diagnosis of venous thromboembolism before further administration of anticoagulant or thrombolytic agents. If the patient's status has deteriorated, bedside diagnostic techniques should be applied to reinforce the suspicion or establish the diagnosis. To restore pulmonary perfusion more rapidly than conventional anticoagulation is suspected to do, several dosing regimens of thrombolytic agents are proposed, with recent interest in short-term thrombolysis and bolus lysis with urokinase or recombinant tissue plasminogen activator. If thrombolysis fails or is contraindicated, catheter embolectomy or surgical embolectomy is indicated. The main therapy is prevention. In this article, clinical assessment, imaging techniques, and therapeutic options described in the published literature are discussed and clinical experiences of an emergency department with a noninvasive diagnostic approach are described.

[Immunopathology and pathogenesis of chronic erysipelas polyarthritis of swine]. / Immunpathologie und Pathogenese der chronischen Rotlauf-Polyarthritis des Schweines

Several immuno-pathological aspects of polyarthritis following experimental infection with erysipelas in pigs were studied for two years. Aseptic and specifically pathogenfree animals were infected subcutaneously and intravenously-intraarticularly with living erysipeals bacteria (erysipelothrix rhusiopathiae) of serotype B. After an initial febrile phase a progressive polyarthritis and disco-spondylitis developed. Some animals also developed thrombo-endocarditis. Hypergammaglobulinemia and high titers of specific antibodies were observed during the whole experimental period. Antiglobulin factors, however, were not detected in the serum or the synovium. In some animals collagen antibodies were demonstrated in synovial tissue. Bacterial examination of the synovium showed that erysipelas bacteria were present in arthritic joints for months. Living erysipelas bacteria were isolated 24 months after the experimental infection from synovial tissue of two pigs. The polyarthritis was characterised by exudates rich in fibrin, villous proliferation, pannus formation, cartilage erosions, and peri-articular fibrosis. IgG and specific erysipelas antibodies were demonstrated in plasma cells from synovial tissue by immuno-histological methods. The findings emphasize the morphological resemblance of the erysipelas induced chronic polyarthritis in pigs to human rheumatoid arthritis.

[Experimental erysipelas in different species as a model for systemic connective tissue disease. II. The chronic phase with special reference to polyarthritis (author's transl)]. / Experimenteller Rotlauf bei verschiedenen Spezies als Modell einer systemischen Bindegewebskrankheit II. Chronische Phase mit besonderer Berücksichtigung der Polyarthritis

INTRODUCTION: In part I of this paper (Schulz et al., 1975) it was shown that in the initial phase of experimental erysipelas a transition from the vascular processes to a systemic connective tissue reaction can be demonstrated in different species. It is the purpose of this paper to describe the chronic phase of the disease with special emphasis on polyarthritis. MATERIALS AND METHODS: 12 spontaneously diseased and 22 experimentally infected pigs were used in experiments to study the pathogenesis of the disease. In addition, 74 Wistar rats and 148 Sprague-Dawley rats were used in the experiments. All experimental animals were specific-pathogen-free and were parenterally infected with the standardized E. insidiosa serotype B strain T 28. The observation period for the pigs was up to 2 years, for the rats up to 11 months. The methods used for pathohistological and electron microscopical studies are described in part I. Immunihistological studies were carried out on synovial tissue with peroxidase-conjugates of goat-anti-pig-IgG, goat-anti-pig-IgM, pig-collagen, E. insidiosa-homogenate and heat-aggregated-pig-IgG. Furthermore, goat-anti-pig-IgG and rabbit-anti-pig-C3 conjugated with FITC were used. Passive hemagglutination tests and Latex agglutination test (Singer and Plotz) were performed to demonstrate rheumatoid factors and collagen antibodies. RESULTS: Polyarthritis occurred in pigs between the 4th and 10th day p.i. and between the 4th and 8th day p.i. in nearly 100% of the infected rats. Fibrinous exudation, proliferation and destruction with pannus formation are marked in most of the joints examined during the first three months. Fibrosis begins 30 days p.i. in the rats' joints and is most severe in both species between the 5th and 8th month. 3 types of lining cells may be differentiated electron microscopically: A (M) cells, B (F) cells and an intermediate form which is found in both species most frequently. Swelling of the endothelial cells together with constriction of the lumen and thickening of the basal membrane occurs in the capillaries. DISCUSSION: A comparison of chronic erysipelas polyarthritis in pigs and rats with rheumatoid arthritis of men reveals many morphological and immunological similarities between the two diseases. Systemic connective tissue activation manifests itself in organs predilected for rheumatic changes, such as heart valves, endocardium and joints. The possible prepetuation of the processes by specific or nonspecific immunomechanisms or by deposits of fibrin is discussed. In addition, experimental erysipelas is reproducible in nearly 100% of the animals given one single subcutaneous application of one defined bacteria strain. Therefore too, erysipelas is suited as an animal model for human rheumatic diseases.