Impatiens parviflora DC. — a natural host of cabbage black ringspot and cucumber mosaic viruses

Identification trials were carried out to determine a virus-like disease ofImpatiene parviflora DC. Two forms of foliar symptoms were observed:

1. a)
   Chiorotie and necrotic areas sometimes fallen out, leaving the leaves “shot holed”;     

Injured mitochondria in cells of Euglena gracilis after DNA gyrase inhibitors treatment

Five quinolone (ofloxacin, cinoxacin, enoxacin, ciprofloxacin, oxolinic acid) and one non-quinolone (coumermycin A1) inhibitors of prokaryotic DNA gyrase used in clinical practice for treatment of bacterial infections were experimentally examinated. As model organism the flagellate Euglena gracilis was used. Ultrastructural changes in chloroplasts and mitochondria caused by inhibitors were quantitavely evaluated. Simultaneously in all cases injury and hereditary loss of chloroplasts (bleaching) were observed in the cells. In some samples about 45% of cup-shaped mitochondria cumulated in the cytoplasm. In damaged mitochondria some degenerative signs were seen, but after the last subcultivation on drug-free media the number of injured mitochondria in the bleached cells yielded to the normal value.     

Expression of CD44 Splice Variants During Lymphocyte Activation and Tumor Progression

Recently, splice variants of CD44 have been described that confer metastatic potential to non-metastasizing rat pancreatic carcinoma and sarcoma cell lines. Using antibodies against variant CD44 (CD44v) sequences, we have examined the expression of variant CD44 glycoproteins on human lymphoid cells and tissues and in colorectal neoplasia. Lymphohematopoietic cells express low levels of CD44v glycoproteins. During the process of lymphocyte activation in vitro and in vivo, expression of CD44v glycoproteins is transiently upregulated. The reaction pattern of various antibodies indicates that these CD44 variants contain the domain encoded by exon v6, which is part of the variant that confers metastatic capability. In human colorectal neoplasia we observed overexpression of CD44 splice variants in all invasive carcinomas. Already at early stages of colorectal tumor progression exon v5 epitopes were overexpressed. Tumor progression was strongly related to expression of CD44 isoforms containing exon v6 encoded domains. The findings establish CD44 variants as tumor progression markers in colorectal cancer.     

On-line immunoanalysis of monoclonal antibodies during a continuous culture of hybridoma cells

The monoclonal-antibody production of an immobilized hybridoma cell line cultivated in a fluidized-bed reactor was monitored on-line for nearly 900 h. The monoclonal antibody concentration was determined by an immuno affinity-chromatography method (ABICAP). Antibodies directed against the product, e.g. IgG, were immobilized on a micro-porous gel and packed in small columns. After all IgG present in the sample was bound to the immobilized antibodies, unbound proteins were removed by rinsing the column. Elution of the bound antibodies followed and the antibodies were determined by fluorescence. The analytical procedure was automated with a robotic device to enable on-line measurements. The correlation between the on-line determined data and antibody concentrations measured by HPLC was linear. A sampling system was constructed, which was based on a pneumatically actuated in-line membrane valve integrated into the circulation loop of the reactor. Separation of the cells from the sample stream was achieved by a depth filter made of glass-fibre, situated outside the reactor. Rapid obstruction of the filter by cells or cell debris and contamination of the sample system was avoided by intermittent rinsing of the sample system with a chemical solution. The intermittent rinsing of the filter, which had a surface of 4.8 cm², resulted in an operational capacity of up to 40 samples (1.0 l total sample volume). Both the sampling system and the analytical device functioned without failure during this long-term culture. The culture temperature was varied between 34 and 40 °C. Raising the temperature from 34 up to 37 °C resulted in a simultaneous increase of growth and specific antibody production rate. Specific metabolic rates of glucose, lactate, glutamine and ammonium stayed constant in this temperature range. A further enhancement of temperature up to 40 °C had a negative effect on the growth rate, whereas the specific monoclonal antibody production rate showed a small increase. The other specific metabolic rates also increased in the temperature range between 38 to 40 °C. This revised version was published online in July 2006 with corrections to the Cover Date.     

Proteinaceous crystals in cells of virus infected plants

Crystal-containing organelles in cells of virus infected plants lying at chloroplasts and mitochondria are identical with single membrane-bound microbodies containing crystals of catalase described in healthy plants. Massive complex inclusions caused by turnip mosaic virus very frequently contain the same microbodies with crystal inclusions; that phenomenon may be related to some pathophysiological changes of virus infected plants. Comparable proteinaceous crystals, but not lying within microbodies limited by a membrane, may also be found in cytoplasm of infected cells. These crystals are sometimes surrounded by a substance resembling the microbody matrix. Disintegrated cytoplasm of virus infected cells may also contain the same crystals lying free in “empty spaces”. Cytopathological effects responsible for this phenomenon and possible artifacts as well are discussed.