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Biomedical subjects

A Baur

Publications and source records attributed to A Baur.

72 records · Page 4Linked to original sources

Synthesis of extracellular matrix components by human ciliary muscle cells in culture.

The production and spatial organization of connective tissue components in ciliary muscle cell cultures was studied with immunohistochemical and ultrastructural methods. Antibodies against collagen types IV and VI, fibronectin and laminin were used. Laminin stains as pericellular network surrounding individual muscle cells. Type IV collagen shows positive cytoplasmic staining and only small foci of extracellular immunofluorescence. Staining for type VI collagen and fibronectin is seen near the ends of the bipolar cells, while the lateral sides of the cells remain unstained. Electronmicroscopy shows that cultured ciliary muscle cells are surrounded by an incomplete basal lamina. In addition, bundles of 5-20 nm thick extracellular microfibrils are seen. The bundles are oriented parallel to the axis of the cells and are in close contact with the cell membrane in areas where membrane-bound dense bands are formed. Immunoelectronmicroscopy indicates that the bundles contain fibronectin and type VI collagen fibrils. While the fibronectin fibrils approach the cell membrane directly, type VI collagen fibrils are usually separated from the cell membrane by fine fibrillous material of different nature. Quality and spatial organization of the extracellular material in ciliary muscle cell cultures shows marked similarities with the extracellular matrix of ciliary muscle in situ.

Adolescent↗

Recombinant antigens in viral diagnosis.

DNA fragments coding for a variety of different viral antigens have been cloned and expressed in Escherichia coli. Selected purified recombinant antigens were used for detection of specific antibodies by means of the ELISA technique. This approach has been used for the development of four different ELISAs for the detection of HIV- and EBV-specific antibodies.

Antibodies, Viral↗

Alpha-lipoic acid is an effective inhibitor of human immuno-deficiency virus (HIV-1) replication.

Alpha-lipoic acid, a naturally occurring disulfide-compound that acts as a cellular coenzyme, inhibits replication of HIV-1 in cultured lymphoid T-cells. Alpha-lipoic acid was added 16 hours after infection of the T-cell lines Jurkat, SupT1 and Molt-4 with HTLV IIIB and HIV-1 Wal (a wild type HIV-1 isolate). We observed a dose dependent inhibition of HIV-1-replication in CPE (Cytopathic effect) formation, reverse transcriptase activity and plaque formation on CD4-transformed HeLa-cells. An over 90% reduction of reverse transcriptase activity could be achieved with 70 micrograms alpha-lipoic acid/ml, a complete reduction of plaque-forming units at concentrations of greater than or equal to 35 micrograms alpha-lipoic acid/ml. An augmentation of the antiviral activity was seen by combination of zidovudine and low dose of alpha-lipoic acid (7 micrograms/ml). Trypan blue staining revealed no toxic effects of alpha-lipoic acids on peripheral blood mono-nuclear cells and T-cell lines even in concentrations of greater than or equal to 70 micrograms/ml. Therefore, we propose the inclusion of alpha-lipoic acid into chemotherapy trials in combination with zidovudine.

Cells, Cultured↗

Cell cultures of human ciliary muscle: growth, ultrastructural and immunocytochemical characteristics.

Primary ciliary muscle cell cultures derived from human donors (16-91 years) were established and characterized by comparing them with ciliary muscle in tissue sections using immunocytochemical and ultrastructural methods. Monoclonal antibodies against desmin, vimentin, alpha-actinin, smooth muscle (sm) specific alpha-actin and von Willebrand factor were used. In tissue sections of the ciliary body, ciliary muscle cells, vascular muscle cells, pericytes, endothelial cells and fibroblasts stain for vimentin. Both types of muscle cells and the pericytes stain for alpha-sm-actin, but only ciliary muscle cells stain for desmin. For tissue cultures, explants of the meridional and partly the reticular portion of the ciliary muscle were dissected and grown directly or after digestion of the explant with collagenase. Ten primary cell cultures with a typical hill-and-valley growth pattern similar to smooth muscle cells and two with a growth pattern similar to fibroblasts were established. All cultures could be subcultured up to the fifth passage. In fibroblast-like cultures 5-10% of the cells stained for alpha-sm-actin. Staining for desmin was not observed. In smooth muscle-like cultures, all cells stained positive for alpha-sm-actin. Desmin staining was not seen in growing non-confluent smooth muscle-like cultures. In confluent cultures, about 10% of the cells stained positive for desmin, preferentially in areas where the cells had formed hills. No culture stained for von Willebrand factor. Staining for alpha-actinin in smooth muscle-like cultures showed that the dense bands of the myofilaments were arranged in register, similar to the typical ciliary muscle cell morphology seen in tissue sections. Ultrastructurally, the smooth muscle-like cultures showed the typical morphology of cultured smooth muscle cells. We conclude that the smooth muscle-like cultures consist of ciliary muscle cells.

Actinin↗

Expression of human immunodeficiency virus (HIV) in naturally infected peripheral blood mononuclear cells: comparison of a standard co-culture technique with a newly developed microculture method.

Peripheral blood mononuclear cells (PBMCs) from 29 patients infected with human immunodeficiency virus (HIV) were cultured by two different methods. One was the standard co-culture technique, the other a newly developed microculture method. In this assay 10(6) PBMCs were cultivated in 250 microliters medium, no activating agents or allogeneic cells were present. P24 antigen production measured by this method was found in 7 out of 11 PBMC cultures of patients in the Walter Reed (WR) stage 1 or 2, whereas only 4 samples were positive by the co-culture procedure. Cultures from patients in the later stages of the disease (WR 5/6) showed a higher p24 production by the co-culture method than by the microculture assay. It is assumed that rapidly growing HIV strains can be better assessed by the co-culture method which may select for these strains. P24 expression can be more easily obtained by the microculture technique even in cases where slowly replicating strains may be present. In conclusion, results from the microculture procedure described may be a useful supplementation to findings observed by the co-culture method.

Adult↗

Antibody- and complement-mediated lysis of HIV-infected cells and inhibition of viral replication.

HIV-1-positive antisera were tested for their ability to lyse HIV-1-infected cells in the presence of active complement. Cytolytic effects caused by sera derived from infected humans were slower than those observed with sera from immunised chimpanzees. Lytic but also negative sera were found among HIV-1-infected asymptomatic men as well as among clinical AIDS cases. Human antisera that lysed infected cells reacted similarly irrespective of whether the complement was heterologous or autologous. Analysis of complement-mediated lysis using defined antisera against recombinant HIV-1 env or core antigens suggested that gp160/gp120 and p24 can act as target antigens for an antibody- and complement-mediated cytolysis of infected cells. Complement alone reduced the spread of HIV-1 infection in CD4+ cells and the ability of HIV-1 and HIV-2 to form plaques in CD4-transfected HeLa cells. Co-operative effects of specific antibodies and complement were the most effective in inhibiting HIV infections.

Acquired Immunodeficiency Syndrome↗

Discrimination of HIV-1 and HIV-2 infection based on recombinant immunoblots.

Recombinant polypeptides representing various parts of structural proteins of HIV-1 and HIV-2 were expressed in E. coli. Fragments of the transmembrane proteins gp41 of HIV-1 (HTLV-IIIB) and gp38 of HIV-2 (LAV-2 ROD) proved to be highly antigenic in the immunoblot test system. Each protein was found to be suitable for detection and differentiation of antibodies in sera of patients infected with HIV-1 or HIV-2. Sera of 100 patients from West-Germany, which were confirmed as positive for HIV-1 antibodies, reacted clearly with the HIV-1 recombinant antigen (fpOE-6); no or only weak immune reactions were seen when the analogous recombinant HIV-2 antigen (fpOE-T) was used in the same immunoblot test. An inverse reaction pattern was found with 7 sera from Africa which, by conventional means, were proved to be HIV-2 antibody positive. These sera specifically reacted with the homologous HIV-2 fusion protein and no or only weakly stained bands were identified on the analogous strip with the HIV-1 antigen (fpOE-6).

Adult↗

Transfer of human and murine globin-gene sequences into transgenic mice.

We have studied the transfer of human and murine globin gene sequences into fertilized mouse oocytes by microinjection. Germline transmission was demonstrated for the human delta- and beta-globin genes contained in the bacteriophage lambda H beta G1. Expression of these human globin-gene sequences was not detectable in either erythroid or nonerythroid tissues. A recombinant plasmid containing the murine beta maj promoter region coupled to the prokaryotic coding sequence for galactokinase was also successfully transferred to two mice, and stable germline transmission of integrated DNA was demonstrated for at least 3 generations. Despite the presence of a murine globin-promoter sequence, expression of the mouse beta maj galactokinase fusion gene was not observed in primary or secondary animals in erythroid or nonerythroid tissues. Analysis of primary and secondary animals from both series of injections revealed extensive de novo methylation in the integrated microinjected DNA. Administration of 5-azacytidine to mice containing the mouse beta maj-promoted galactokinase gene resulted in partial hypomethylation was associated with an apparent two- to threefold increase in galactokinase (gal K) gene expression.

Animals↗

Ethylene production from propanal.

Tracer studies using a model system consisting of Cu(2+) and ascorbate indicate that carbons 2 and 3 of propanal are converted to ethylene, and carbon 1 is converted to formic acid and CO(2). A mechanism accounting for this reaction is described. In apple tissue, methionine but not propanal is readily incorporated into ethylene. It is therefore concluded that propanal is not a precursor of ethylene in this fruit.

Journal Article↗

MR-imaging changes of musculoskeletal soft-tissue sarcomas associated with neoadjuvant chemotherapy and hyperthermia.

PURPOSE: To evaluate early changes in musculoskeletal soft-tissue sarcomas under neoadjuvant chemotherapy combined with regional hyperthermia (RHT). PATIENTS AND METHODS: Nineteen consecutive patients with high-grade soft-tissue sarcomas of the musculoskeletal system were treated with neoadjuvant chemotherapy combined with RHT. Patients were imaged, using a high field MR-scanner, before onset of therapy, immediately after one and after four cycles of therapy. The images were evaluated for volume reduction and development of tumour necrosis. In addition, side effects such as surrounding soft-tissue oedema, bleeding and muscle or bone marrow necrosis were analysed. RESULTS: Tumour volume reduction was significant after the completion of neoadjuvant therapy (mean 49%, range 5-91%; (p < 0.001). Extent of tumour necrosis was also significantly different before (mean 22%) and after therapy (mean 58%, p < 0.001). Three patients showed strong tumour necrosis already after one cycle of treatment. Tumour volume reduction was not associated with the extent of pre-existing necrosis or necrosis development. The extent of tumour volume before start of therapy did not affect volume reduction or necrosis induction after therapy. Reduction of tumour oedema was significant after therapy (p < 0.001). No side effects were observed during thermochemotherapy. CONCLUSION: Neoadjuvant chemotherapy combined with RHT resulted in significant tumour volume reduction and induction of tumour necrosis, which can be detected early and monitored closely with MRI.

Adult↗

Half-body radiotherapy. Evaluation of the technique in normal dogs.

Eight healthy mongrel dogs were treated with half-body irradiation (HBI) in a pilot study to evaluate the technique and radiotolerance of different organs. Cranial and caudal half-body fields were established using the 13th thoracic vertebra as the dividing point. Under general anesthesia, either 7 or 8 Gray (Gy) were delivered to one half of the body using opposing radiation portals. The other half of the body was similarly treated 28 days later. The dogs were monitored for 12 months. Significant radiation effects included transient bone marrow suppression and radiation sickness. There were no serious or life-threatening problems, but the 8 Gy group consistently showed more severe clinical signs and histologic changes than the 7 Gy group. Total body irradiation in two fractions of 7 or 8 Gy given 1 month apart appears to be a safe treatment that can be developed for therapy in veterinary oncology.

Animals↗

Half-body radiotherapy in the treatment of canine lymphoma.

In a Phase I-II study, half-body radiotherapy was used to treat 14 dogs with multicentric lymphoma. Using this technique, a radiation dose of 7 Gray (Gy) was delivered to one half of the body in a single exposure. The other half of the body was treated approximately 28 days later. Of 14 treated dogs, 11 (79%) had a measurable decrease in tumor size. Five dogs achieved a complete or partial remission with a mean duration of 102 and 54 days, respectively. In predicting response to therapy, poor prognostic factors included large tumor burdens, advanced disease stage, and chemotherapy-resistant tumors. Side effects of treatment were divided chronologically into acute (radiation sickness, tumor lysis), subacute (bone marrow suppression), and chronic (radiation pneumonitis, lymphoma-cell leukemia) syndromes. Complications were more severe in tumor-bearing dogs when compared with healthy control animals. Dogs with small tumor burdens and minimal internal disease had fewer complications compared with those with more advanced disease.

Animals↗