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Biomedical subjects

Dennis Tappe

Publications and source records attributed to Dennis Tappe.

10 recordsLinked to original sources

Characterisation of EmMPK1, an ERK-like MAP kinase from Echinococcus multilocularis which is activated in response to human epidermal growth factor.

Mitogen-activated protein (MAP) kinases are key regulators of cellular signalling systems that mediate responses to a wide variety of extracellular stimuli and should also play a central role in developmental mechanisms of parasitic helminths. Until now, however, no MAP kinase orthologue has been characterised in a member of this parasite group. Here, we report the identification and characterisation of such a molecule, EmMPK1, from the human parasitic cestode Echinococcus multilocularis. Using a degenerative PCR approach, we isolated and completely sequenced the 1.2kb cDNA for EmMPK1 which displays significant homologies to known MAP kinases of different phylogenetic origin. EmMPK1 contains all amino acid residues which are characteristic for MAP kinases, including a conserved TEY motif which identifies the protein as a member of the ERK subfamily of MAP kinases. The corresponding gene, emmpk1 (6.9 kb), was characterised and contained 10 introns. Southern blot hybridisation studies showed that emmpk1 is present as single copy locus in E. multilocularis. Using RT-PCR analyses we demonstrated that emmpk1 is expressed in form of three different transcripts which derive from alternative splice acceptor site utilisation at intron 9. Using EmMPK1-specific antibodies in Western blot studies and immunohistochemistry, we detected the Echinococcus protein and its phosphorylated form in the larval stages metacestode and protoscolex during in vitro cultivation and during an infection of the intermediate host. EmMPK1, immunoprecipitated from Echinococcus lysate, was able to phosphorylate myelin basic protein in activity assays, indicating that it is a functionally active MAP kinase. Finally, we also show that phosphorylation of EmMPK1 is specifically induced in vitro-cultivated E. multilocularis metacestode vesicles in response to exogenous host serum and upon addition of human epidermal growth factor. These data indicate that the E. multilocularis metacestode is able to sense epidermal growth factor from the host which results in an activation of the parasite's MAP kinase cascade.

Alternative Splicing↗

Central nervous system borreliosis mimicking a pontine tumour.

In childhood, facial nerve palsy and headache are typical symptoms of second and third stage neuroborreliosis. While focal demyelination is occasionally observed on MRI scans, the appearance of a tumorous lesion is extremely rare. The case of a 10-year-old girl with neuroborreliosis mimicking a space-occupying lesion in the brainstem, without any previously recognized manifestations of borreliosis, is reported.

Anti-Bacterial Agents↗

Subcutaneous Taenia crassiceps infection in a patient with non-Hodgkin's lymphoma.

Infections with larvae of Taenia crassiceps are rare in humans and have mostly affected patients with acquired immunodeficiency syndrome. We report the first case of a patient with malignancy (non-Hodgkin's lymphoma) and infection of the subcutis and muscles of the hand and forearm. Surgery and antiparasitic chemotherapy led to a complete cure.

Aged, 80 and over↗

Fatal Clostridium tertium septicemia in a nonneutropenic patient.

Clostridium tertium septicemia is a rare condition that predominantly occurs in neutropenic patients with concomitant abdominal disease. We report the fatal case of a nonneutropenic, 51-year-old patient with mechanical ileus and post-operative C. tertium septicemia, resulting in widespread pathology with multi-organ failure. As C. tertium is aerotolerant, often gram-variable and mostly resistant to broad-spectrum cephalosporins, differentiation is difficult and empirical therapeutic strategies may fail.

Anti-Bacterial Agents↗

Molecular cloning and characterization of Ras- and Raf-homologues from the fox-tapeworm Echinococcus multilocularis.

To better understand growth regulation in the human parasitic cestode Echinococcus multilocularis, we have cloned and characterized the parasite's orthologues of the key regulatory factors Ras and Raf. Using a degenerative PCR approach a gene, emras, was identified whose gene product, EmRas, showed high homology (79% identical residues) to human Ras and contained all amino acid residues which are characteristic for this subfamily of small GTPases at the corresponding positions. Recombinantly expressed EmRas bound GTP and was farnesylated, but not geranyl-geranylated, by Echinococcus lysate in an in vitro prenylation assay. Furthermore, upon expression in yeast, emras was able to functionally complement the Saccharomyces orthologue RAS2 in an invasive growth assay. In Western blot analyses using an anti-EmRas antibody, the Echinococcus factor could be detected in lysates of the larval stages metacestode and protoscolex. By immune-histochemistry, EmRas was shown to localize to the germinal layer of the metacestode and to tegumental structures of the protoscolex, particularly around the rostellum and the sucker regions. In addition, we fully characterized the gene emraf whose product, EmRaf, displayed considerable homology to mammalian Raf-kinases and orthologous factors from Drosophila and Caenorhabditis elegans. emraf was co-expressed with emras in the larval stages metacestode and protoscolex during in vitro cultivation and during an infection of the intermediate host as assessed by RT-PCR experiments. The emraf gene was composed of nine exons and eight introns and shared four highly conserved exon-intron boundaries with the human gene encoding Raf-1, suggesting that both genes derived from a common evolutionary ancestor. Southern blot hybridizations demonstrated that emraf is a single copy gene. Using the yeast two-hybrid system, EmRaf was shown to interact with EmRas, but not with EmRal, a previously characterized orthologue of mammalian Ral GTPases. This is the first characterization of a Ras orthologue from a cestode and the first report on a Raf-like kinase from a platyhelminth. The data presented herein will form a solid basis for further investigations on Echinococcus signaling systems that are involved in growth control and development of the parasite.

Amino Acid Sequence↗

Long-term in vitro cultivation of Echinococcus multilocularis metacestodes under axenic conditions.

We report here on the development of an in vitro system for the long-term cultivation of Echinococcus multilocularis larvae under axenic conditions. In the absence of feeder cells from the host, long-term survival of the parasite depended strictly on low oxygen conditions and the presence of reducing agents in the medium. Host serum supported survival of the parasite but the growth of metacestode vesicles and differentiation towards the protoscolex stage only occurred in the presence of culture medium that was preconditioned by hepatoma cells or several other immortal cell lines. On the basis of this in vitro system, future analyses on the identification of host-derived growth factors for E. multilocularis will be greatly facilitated.

Animals↗

Osteomyelitis and skin ulcers caused by Mycobacterium szulgai in an AIDS patient.

Mycobacterium szulgai is a ubiquitious non-tuberculous mycobacterium causing infection in immunocompetent and immunocompromized patients. Clinically mimicking pulmonary tuberculosis in most cases described, rarely other manifestations occur. Here we report the case of an AIDS patient with osteomyelitis of the hand and toe, accompanied by multiple cutaneous ulcers of the chest and forearm. The case highlights the unusual combination of osteomyelitis and skin ulcers without pulmonary infection and describes the likely cutaneous route of infection in a patient who keeps tropical fish.

Acquired Immunodeficiency Syndrome↗

Long-term benzimidazole treatment of alveolar echinococcosis with hematogenic subcutaneous and bone dissemination.

We report a case of advanced alveolar echinococcosis (AE) that presented like a malignant tumor. It was diagnosed histologically from a subcutaneous nodule with skin inflammation on the right leg. Additionally, the patient showed bone metastases in the lower thoracic spine and the left third toe. This is the first case with proven hematogenic spread of AE to a subcutaneous site. The patient was treated with albendazole and remained stable for 6 years. When progression of AE occurred the therapy was changed to mebendazole, resulting in a stable condition for further 4 years.

Albendazole↗