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.     

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.     

A negative influence of manure on calcareous soils

Summary On a soil rich in CaCO₃ in a semiaride climate in Algeria a hard soil layer, impermeable for plant roots, was formed in a depth of 20–40 cm after farmyard manure application and irrigation. To find the reason soil samples of this field were taken and leaching experiments were carried out in the laboratory, with the result that much more Caions and HCO₃-ions were leached out of the soils with farmyard manure application than from the soils without manure. Probably the high amount of CO₂, being liberated by the organic matter, and the irrigation water dissolved the CaCO₃ in the soil, and the formed Ca- and HCO₃-ions followed the movement of water in the soil. Where the Ca(HCO₃)₂ reached soil layers with a less amount of CO₂, CaCO₃ precipitated and formed the hard soil layer. In order to avoid the formation of such calcareous crusts on irrigated, limy soils in a dry climate it is recommended to fertilize rather often small quantities instead of rarely big quantities of farmyard manure.     

Bacterial mesosomes: Real structures of artifacts?

The ultrastructural study of membrane organization in gram-positive bacteria related to the OsO₄ fixation conditions revealed that large, complex mesosomes are observed only when the bacteria are subjected to an initial fixation with 0.1% OsO₄ in the culture broth, as in the prefixation step of the Ryter-Kellenberger procedure. Evidence was obtained suggesting that the large mesosomes are produced by this prefixation. The kinetic study of the membrane morphological alterations occurring during the prefixation of Bacillus cereus with 0.1% OsO₄ in the culture broth showed that the amount of mesosome material increases linearly from zero to a maximum observed at 1.7 min of prefixation and that at about this time a maximum is reached for the number of mesosomes per unity of cell area and for the average individual mesosome area. The large mesosomes observed in gram-positives fixed by the complete Ryter-Kellenberger procedure would be the result of the membrane-damaging action of 0.1% OsO₄. Such damaging action was deduced from the observation that 0.1% OsO₄ quickly lyses protoplasts and induces a quick and extensive leakage of intracellular K⁺ from B. cereus and Streptococcus faeculis. In support of that interpretation is the observation that in bacteria subjected to several membrane-damaging treatments, mesosome-like structures are seen after three different fixation procedures. In bacteria initially fixed with 1% OsO₄, 4% OsO₄ or 2.5% glutaraldehyde, no large, complex mesosomes are observed, small and simple invaginations of the cytoplasmic membrane being present. The size of these minute mesosomes is inversely proportional that causes of fixation. Uranyl acetate was found among the studied fixatives the one to the rate the least damage to bacterial membranes. This fixative satisfactorily preserves protoplasts. In bacteria initially fixed with uranyl acetate no mesosomes were found. The results of the present work throw serious doubts on the existence of mesosomes, both large and small, as real structures of bacterial cells. It is proposed that a continuous cytoplasmic membrane without infoldings (mesosomes) would be the real pattern of membrane organization in gram-positives.