Effects of thalidomide on the local Shwartzman reaction in mice and rabbits.

The Shwartzman reaction is an animal model displaying histopathological vasculitis phenomena. Extravasation and swelling due to increased vascular permeability and cellular infiltration, which are hallmarks of the Shwartzman reaction, were evaluated as leakage of i.v.-injected Evans Blue dye and by histological and immunohistological characteristics in rabbits and mice. (+/-)-Thalidomide, (-)-thalidomide, (+)-thalidomide and dexamethasone inhibited the increase of vascular permeability in the local Shwartzman reaction. Histologically, the intensity of the Shwartzman reaction was reduced. In mice thrombus formation and leukocytoclastic vasculitis was inhibited by (+/-)-thalidomide and (+)-thalidomide. ICAM-1 expression was markedly reduced after (+)-thalidomide injection. Thalidomide and dexamethasone pretreatment reduced Mac-1 expression on perivascular infiltrated granulocytes. The inhibitory effect of thalidomide on vasculitis of the Shwartzman reaction may thus be related to reduction of adhesion molecule expression.

The experimental toxicology of tramadol: an overview.

The experimental toxicological findings of tramadol are reviewed and discussed. Tramadol is a centrally acting analgesic. In acute toxicity studies, LD50 values are estimated to be around 300-350 mg/kg body weight (rat, mouse, oral administration). After intravenous administration the LD50 values ranged from 50 to 100 mg/kg body weight. In subacute and chronic toxicity studies, clinical signs of intoxication are mainly behavioural disorders and convulsions, beginning at dose levels of 25 mg/kg. Clinical pathological alterations or morphological lesions, in particular neuropathological findings were not detected. Overall, the battery of mutagenicity studies shows no evidence of a genotoxic risk to man. Reproductive and developmental toxicity investigations and carcinogenicity studies were without substance-dependent findings. Toxicological and toxicokinetical data of both enantiomers did not show biologically relevant deviations in comparison to the data on tramadol. The toxicological characteristic of this compound is demonstrated.

Immunohistological detection of bovine viral diarrhoea virus antigen in the central nervous system of persistently infected cattle using monoclonal antibodies.

In a total of 25 cattle persistently infected with bovine viral diarrhoea virus (BVDV) the distribution of viral antigens in the central nervous system was studied. Using a panel of monoclonal antibodies (anti pestivirus C16; anti cytophathic BVDV C38; anti cytopathic and non-cytopathic BVDV C42; anti gp53 BVDV CA-1 and CA-3) and the indirect immunoperoxidase technique, BVDV antigen was located exclusively in neurons. Predilection sites for viral persistence were cerebral cortex and hippocampus. Morphological cellular alterations were not seen. Reactive perivascular lymphocytic infiltrations were occasional findings.

The beagle dog as a predictor of gastrointestinal findings in humans caused by the prostacyclin analogue taprostene.

Beagle dogs were exposed orally to the prostacyclin analogue taprostene for four weeks. Dose levels of 200-3000 micrograms/kg body weight/day were used. Specific activity of taprostene on the digestive system compared to other species is reported. It is characterized by hypermotility of the gastrointestinal tract resulting in intestinal invagination in some animals. Gastrointestinal symptoms occurred also after intravenous administration indicating a systemic stimulating effect on smooth muscles. Concerning reversible gastrointestinal side effects in humans after intravenous infusion of prostacyclin the results of this subacute toxicity study indicated that the dog is an adequate and sensitive species for preclinical testing of prostacyclins.

Local skin reactivity after induction of Shwartzman reaction in rabbits.

A local Shwartzman response was elicited in rabbits by an intradermal injection of the Salmonella typhosa endotoxin lipopolysaccharide (LPS) followed 24 hours later by an intravenous challenge injection with zymosan. After the intravenous challenge, necrotizing vasculitis developed in the prepared skin sites which was characterized by microthrombi, accumulation of neutrophil granulocytes, fibrin deposition and extravasation of red blood cells. Evans' blue extravasation into the altered tissue was significantly reduced, and histologically, the intensity of the Shwartzman reaction in the skin was reduced by pretreatment with thalidomide and dexamethasone. The mechanism of reduction of an LPS-induced local Shwartzman reaction by thalidomide is discussed.

Metastasizing seminal vesicle adenocarcinoma in a Wistar rat.

An adenocarcinoma in the seminal vesicles of a 15-month-old male Wistar rat from a 30-month inhalation study is described. The rat was killed because of cachexia, apathy and a large palpable mass in the abdominal cavity. Macroscopic examination of the abdominal cavity revealed a 3.8 cm x 3.2 cm yellow-grey to pink mass, firm to soft in consistency. The cut section revealed cystic spaces. Histologically, the mass consisted of epithelial cells arranged in glandular and solid patterns with abundant amounts of connective tissue. Epithelial tumour cells were round-to-cylindrical with round-to-oval basophilic nuclei and one or two prominent nucleoli and a distinct eosinophilic cytoplasm. The glandular structure contained clusters of macrophages in their lumen with eosinophilic cytoplasm and indented nuclei. Extensive necrosis and reactive inflammation were present. The histological features of the small nodules in the pancreas and on the surface of the liver, rectum and urinary bladder resembled those of the primary tumour in the seminal vesicles. Based on these criteria, the neoplasm (mass) was diagnosed as an adenocarcinoma of the seminal vesicles. The immunohistological examination confirmed the diagnosis, i.e. immunostaining was positive for cytokeratins (4, 7, 14, 15, 18, and 19), vimentin, PCNA, and ED(1).

Endocardial schwannomas in the Wistar rat.

Endocardial neoplasms were observed in 13 Wistar rats, 15-30 months old, from a 30-month inhalation study. Eight of the rats were dissected at 30 months and the other five between 15 and 29 months. Histologically, the lesions in the heart were diagnosed as endocardial schwannomas. Two of the 13 schwannomas were malignant, while the rest were benign. One malignant schwannoma metastasized to the thymus and the other metastasized to the aorta, oesophagus, liver and spleen. Some schwannomas appeared as early lesions limited to the endocardium consisting of round to ovoid cells interspersed with spindle cells, while other schwannomas appeared as advanced lesions consisting of a thin superficial layer of round to ovoid cells and a deeper layer of spindle cells characteristic of endocardial schwannomas. The histological appearance of the metastases was identical to that of the primary tumour. The diagnosis was confirmed by positive immunostaining for S-100 protein. The incidence of endocardial schwannomas in the Wistar rat strain was 1.8%.

Intestinal inflammation in TNBS sensitized rats as a model of chronic inflammatory bowel disease.

An enteritis, based on a delayed-type hypersensitivity reaction, was induced in TNBS (2,4,4-trinitrobenzenesulphonic acid) sensitized rats by multiple intrajejunal challenge with TNBS via an implanted catheter. This treatment induced chronic inflammation of the distal small intestine characterized by intense hyperaemia, oedema and gut wall thickening as assessed by macroscopic scoring and weighing a defined part of the dissected intestine. Histologically, the inflammatory response included mucosal and submucosal cell infiltration by lymphocytes and histiocytes, transmural granulomatous inflammation with multinucleated cells and activated mesenteric lymph nodes. Ex vivo stimulated release of the inflammatory mediator LTB(4) in the dissected part of the intestine was increased following TNBS treatment. Drug treatment with sulphasalazine or 5-aminosalicylic acid improved the enteritis score and attenuated TNBS induced oedema formation and LTB(4) production. The applicability and relevance of this new model are discussed with respect to drug development and basic research of inflammatory bowel diseases.

Metastasizing fibrosarcoma in a Wistar rat--case report.

A 19-month-old male control Wistar rat from a 30-month inhalation study showed a subcutaneous greyish-white mass extending from the throat to the thoracic cavity. The rat had been euthanized because of its poor general condition. Histologically, the mass was diagnosed as a fibrosarcoma infiltrating the masseter muscle with metastases in the lungs, liver and heart. The primary tumour was characterized by fusiform spindle cells producing various amounts of interlacing bundles of collagen. The cells formed a characteristic herringbone pattern and mitotic figures were frequent. The histological parameters of the metastases were practically identical to those seen in the primary tumour. The diagnosis was confirmed by trichrome staining and positive immunostaining for vimentin and was differentiated from leiomyosarcomas by its negative immunostaining for desmin, from schwannomas by its negative immunostaining for S-100 and from malignant fibrous histiocytomas by the absence of giant cells. The incidence of fibrosarcomas in Wistar rats is very low (up to 3%) and metastasis is rarely observed.

Hepatocholangiocellular carcinoma in a rat--case report.

A mixed epithelial tumour in the liver of a 24-month-old male Wistar rat from a 30-month inhalation study is described. The rat, which was in a group exposed to low concentrations of diesel exhaust, was euthanized because of emaciation, forced respiration and abnormal gait. Macroscopic examination of the enlarged liver revealed multiple partly confluent beige-red nodules up to 1.5 cm in diameter. Small nodules up to 7 mm in diameter were seen in the spleen. Histologically, the tumour nodules in the liver consisted of hepatocellular and cholangiocellular components. The hepatocellular component consisted of moderately differentiated polygonal to round hepatocytes about twice as large as normal hepatocytes and having hyperchromatic, centrally located nuclei with prominent nucleoli and eosinophilic cytoplasm. Foci of haematopoiesis and focal necroses were prominent. The cholangiocellular component was moderately differentiated and consisted of tubular structures lined by low cuboidal to cylindrical cells showing cytoplasmic basophilia and small dark nuclei without prominent nucleoli. The histological features of the nodules in the spleen corresponded to those of the primary tumour in the liver. Based on these criteria, the tumour nodules were diagnosed as hepatocholangiocellular carcinoma. The immunohistological examination confirmed the diagnosis, i.e. immunostaining for cytokeratins was positive for eight and 18 (hepatocellular carcinoma) and for seven and 19 (cholangiocellular carcinoma) as well as for vimentin (dense fibrous stroma). This tumour is considered to be spontaneous because of its single occurrence.

Immunohistological detection of Saruplase (recombinant single-chain urokinase-type plasminogen activator) in normal rat tissue.

Saruplase--a recombinant single-chain urokinase-type plasminogen activator was identified immunohistochemically in normal rat tissue after intravenous administration by means of a polyclonal antibody. For this purpose, rat tissues were fixed in various ways (liquid nitrogen, ethanol, formaldehyd solution). Saruplase could be detected by the PAP method, streptavidinbiotin system and indirect immunofluorescence in the kidney (proximal tubule), liver (hepatocytes, Kupffer cells) and spleen (reticular cells). Saruplase was not localized in the rat endothelium. It is discussed that the rat-specific receptors for urokinase-type plasminogen activator on endothelial cells cannot bind Saruplase due to the extreme species specificity.

Distribution of bovine virus diarrhoea viral antigens in the central nervous system of cattle with various congenital manifestations.

Distribution of bovine viral diarrhoea virus (BVDV) antigens in the central nervous system (CNS) of 26 cattle persistently BVDV infected, 11 cattle with mucosal disease (MD), and 32 calves with congenital brain malformations was studied using monoclonal antibodies against BVDV epitopes. In persistently infected cattle and in cattle with MD, a widespread infection of neurons was present. Predilection sites for BVDV antigens were the cerebral cortex and the hippocampus. In calves with congenital encephalopathies, viral antigen-containing neurons could only be detected in the CNS of four animals. From the topographical distribution of BVDV antigens in these four postnatal cases with end-stage lesions, no conclusions could be drawn concerning the pathogenesis of BVDV-induced encephalopathies.

Inhibitory effects of thalidomide on cellular proliferation, endoneurial edema and myelin phagocytosis during early wallerian degeneration.

In addition to the well-known teratogenic effect of thalidomide, previous studies have revealed mild immunosuppressive properties and, more recently, an anti-angiogenic activity. To find out more about the specificity of these effects we studied the influence of orally administered thalidomide on Wallerian degeneration in rats. Wallerian degeneration is a potent experimental model for studying reproducible cell proliferation in vivo. Examination of distal nerve segments of transected sciatic nerves from rats that had been treated with thalidomide (2 x 250 mg/kg per day) revealed a significant reduction of endoneurial cell counts at 10-15 days after surgery compared to that seen in controls. This effect was not statistically significant, at a very early stage of Wallerian degeneration, i.e., at 5 days after transection of the nerve. Subperineurial edema and phagocytosis was also reduced, although this was not statistically significant. This apparently nonspecific inhibitory effect of thalidomide during early Wallerian degeneration shown in the present study should be investigated further for its possible relationship to other previously established inhibitory activities of thalidomide, especially its immunosuppressive effect in man.