Adenoviral transduction supports matrix expression of alginate cultured articular chondrocytes.

The present study examines the effects of adenoviral (Ad) transduction of human primary chondrocyte on transgene expression and matrix production. Primary chondrocytes were isolated from healthy articular cartilage and from cartilage with mild osteoarthritis (OA), transduced with an Ad vector and either immediately cultured in alginate or expanded in monolayer before alginate culture. Proteoglycan production was measured using dimethylmethylene blue (DMMB) assay and matrix gene expression was quantified by real-time PCR. Viral infection of primary chondrocytes results in a stable long time transgene expression for up to 13 weeks. Ad transduction does not significantly alter gene expression and matrix production if chondrocytes are immediately embedded in alginate. However, if expanded prior to three dimension (3D) culture in alginate, chondrocytes produce not only more proteoglycans compared to non-transduced controls, but also display an increased anabolic and decreased catabolic activity compared to non-transduced controls. We therefore suggest that successful autologous chondrocyte transplantation (ACT) should combine adenoviral transduction of primary chondrocytes with expansion in monolayer followed by 3D culture. Future studies will be needed to investigate whether the subsequent matrix production can be further improved by using Ad vectors bearing genes encoding matrix proteins.

Effect of betaTCP filled polyetheretherketone on osteoblast cell proliferation in vitro.

Non-resorbable thermoplastic polymers have become more important for reconstructive surgery due to their excellent chemical and physical properties. Polyetheretherketone-beta-tricalcium phosphate (betaTCP-PEEK) composites were developed as alternative materials for load-bearing applications. This study presents the effect of polyetheretherketone (PEEK) specimens incorporated with 5, 10, 20 and 40 wt% beta-tricalcium phosphate (betaTCP) and processed by injection molding on cultivated osteoblast cells. Normal human osteoblast (NHOst) cells were seeded onto polymer discs to evaluate cell viability and proliferation after 24, 72 and 120 h of cultivation by employing the WST-1 assay. Standard tissue culture plastic was used as a control. The osteoblast cells were found to be viable in all PEEK groups, while the cell proliferation was progressively inhibited due to the incorporated beta-tricalcium phosphate. BetaTCP-PEEK showed concentration independent decrease of cell proliferation compared to the unfilled PEEK and the control group. In summary, this study confirms the non-toxic nature of pure PEEK, whereas this could not definitely be verified for betaTCP-PEEK as a composite material in chosen concentrations of beta-tricalcium phosphate in vitro.

Rabies in red foxes (Vulpes vulpes) experimentally infected with European bat lyssavirus type 1.

The susceptibility of red foxes (Vulpes vulpes) to European bat lyssavirus type 1 (EBLV-1) infection was examined. Eight foxes were inoculated intramuscularly (i.m.) with 10(4.9) foci-forming units (FFU) (n = 4) and 10(5.1) FFU (n = 4) and observed for up to 90 days. All foxes showed manifestations of a neurologic disorder (e.g. seizures, myoclonus, agitation), starting as early as 5 days post-infection (p.i.). Subsequently, all animals showed improvement followed by one or more relapses. One fox was killed 3 days after it recovered, 26 days post-infection. Two other foxes were also killed 38 and 54 days post-infection after severe neurologic signs returned. All foxes developed a humoral immune response against EBLV-1 as determined in serum and brain tissues. However, no rabies virus antigen was detected in the brain, other tissues and secretions examined (e.g. salivary gland, saliva, tonsils, lungs) by using different standard diagnostic techniques [fluorescent antibody test, reverse transcription polymerase chain reaction (RT-PCR), rabies tissue culture inoculation test], with the exception of one fox in which EBLV-1 RNA was detected by RT-PCR in only the spinal cord. Brain tissues showed moderate to severe multifocal, mononuclear encephalomyelitis in the three foxes that were killed during the observation period, although no EBLV-1 virus was detectable in these tissues.

[Alterations of hemostasis in liver cirrhosis of the dog]. / Veränderungen der Hämostase bei der Leberzirrhose des Hundes.

In seven dogs with histologically proven liver cirrhosis the activity of the single coagulation factors with the exception of factor VIII:C, of the inhibitors antithrombin III and protein C as well as plasminogen and alpha 2-antiplasmin was distinctly lower than in the control group (p < 0.0001). The changes of the factors VII [median (x0.50) = 17 %] and X (x0.50 = 18 %) as well as of protein C (x0.50 = 15 %) were particularly pronounced. Diminution of activity certainly exceeded also in nearly all of the remaining haemostatic proteins the decrease of albumin concentration. Besides the shorter half life time, this reflected an increased consumption in consequence of intravascular coagulations and fibrinolysis. The latter could also be seen from the significantly increased concentrations of soluble fibrin and fibrin(ogen) degradation products. Therefore, the alterations of the haemostatic system measured in dogs in many details were in accordance with findings in human beings suffering from liver cirrhosis.

[Examination of primary SPF swine after experimental infection with Haemophilus parasuis. Clinical symptoms, changes in hematological parameters and in the parameters of the cerebrospinal fluid]. / Untersuchungen an primären SPF-Ferkeln nach experimenteller Infektion mit Haemophilus parasuis. Klinische Symptomatik, Veränderungen von hämatologischen Parametern und von Parametern des Liquor cerebrospinalis.

The objectives of this work were to cause the Glässer's disease (GD) in primary specific pathogen free piglets after experimental infection, to observe the clinical symptoms and to examine the influence of the infection on the haematological parameters. GD was caused by experimental infection of Haemophilus parasuis in seven to eight weeks old specific pathogen free piglets. In relation to the infection route the morbidity was high (83-100%) and 20% of the infected piglets died. Based on the physical examination fever, respiratory distress, cramps and paralysis were observed which are typical for GD. Arthritis and nerval symptoms are also typical but less common in Glässer's disease. PCV was significantly decreased and WBC significant increased before the piglets were euthanatized.

[Hepatitis B virus infections in hospitals]. / Infektionen mit Hepatitis-B-Virus im Krankenhaus.

Subclinical and clinical cases of hepatitis B virus (HBV) infection in hospital personnel were recorded over a five-year period. Although our data indicate the existence of "high-risk areas" like laboratories, dialysis units or surgical wards, it was found that 50% of all HBV infections were observed outside these areas. During the same period, most of our inpatients were monitored for HBs-antigenemia. It could be demonstrated that more than 50% of all cases of HBs-antigenemia were not known before and were treated primarily for other reasons than for hepatitis B. As a consequence from our data and from the screening for environmental HBs-antigen on laboratory surfaces it may be concluded for the hospital routine: Even minimal contaminations with blood should be avoided, all personnel should be vaccinated against HBs-antigen, HBs-antigen should be determined in every inpatient to detect inapparent virus carriers.