Chemotactic assay for biological effects of silver nanoparticles

A method is proposed for assessing the biocidal efficacy of water-dispersed nanoparticles of silver. It is based on negative chemotaxis of the plasmodia of the slime mold Physarum polycephalum. Biocidal and repellent effects were compared for silver nanoparticles, Ag⁺ ions, and AOT in solution and in the agar gel. In such characteristics as increasing the period of auto-oscillations of contractile activity, decreasing the area of spreading on substrate, and substrate preference in spatial tests, silver nanoparticles proved to be substantially more effective than Ag⁺ and AOT. The lethal concentrations of the nanoparticles were close to those found earlier for bacteria and viruses. The chemotactic tests allow quantitative assessment of the biological reaction and monitoring its dynamics; in resolution, they are superior to the tests based on the lethal action of biocidal agents.     

Isolation and characteristics of Thiopedia rosea (neotype)

Four new strains of Thiopedia rosea were isolated in pure culture from blooms of platelet-forming purple sulfur bacteria in the top layers of the anoxic hypolimnia of two freshwater lakes. Individual cells of the new strains as well as of T. rosea strain 4211 were spherical to oval, nonmotile and contained gas vesicles in the central part. The predominant photosynthetic pigments were bacteriochlorophyll a and okenone. All strains were strictly anaerobic and obligately phototrophic. Optimal growth occurred at low light intensities of 100 E · m^-2 s^-1 (tungsten lamp); intensities above 150 E · m^-2 s^-1 inhibited growth completely. Photoautotrophic growth was possible at sulfide concentrations up to 0.6 mM; higher concentrations were inhibitory. Acetate, butyrate and valerate supported photoorganotrophic growth in the presence of bicarbonate and sulfide concentrations below 1 mM. Sulfide was required as a source of cellular sulfur because assimilatory sulfate reduction is lacking. All strains were assigned to the species Thiopedia rosea with strain 4211 as a neotype.Dedicated to Prof. Dr. H. G. Schlegel on the occasion of his 66th birthday     

Characterization of a calcium/calmodulin-dependent protein kinase homolog from maize roots showing light-regulated gravitropism

Roots of many species respond to gravity (gravitropism) and grow downward only if illuminated. This light-regulated root gravitropism is phytochrome-dependent, mediated by calcium, and inhibited by KN-93, a specific inhibitor of calcium/calmodulin-dependent protein kinase II (CaMK II). A cDNA encoding MCK1, a maize homolog of mammalian CaMK, has been isolated from roots of maize (Zea mays L.). The MCK1 gene is expressed in root tips, the site of perception for both light and gravity. Using the [³⁵S]CaM gel-overlay assay we showed that calmodulin-binding activity of the MCK1 is abolished by 50 M KN-93, but binding is not affected by 5 M KN-93, paralleling physiological findings that light-regulated root gravitropism is inhibited by 50 M KN-93, but not by 5 M KN-93. KN-93 inhibits light-regulated gravitropism by interrupting transduction of the light signal, not light perception, suggesting that MCK1 may play a role in transducing light. This is the first report suggesting a physiological function for a CaMK homolog in light signal transduction.Abbreviations CaM calmodulin - CaMK (II) Ca²⁺/calmodulin-dependent protein kinase (II) - CBP CaM-binding protein - CDPK Ca²⁺-dependent protein kinase - MCK1 maize homolog of mamalian CaMK This work is supported by the National Aeronautics and Space Administration grant No: NAGW 238.     

The Casparian strip in pea epicotyls: Effects of light on its development

The Casparian strip, which is specific to roots, was studied in the epicotyls of dark-grown seedlings of pea (Pisum sativum L.) where it was found to have the same morphology and properties as the strip in roots. In dark-grown seedlings, the distance between the upper-most position of the Casparian strip and the bending point of the hook (about 37 mm) did not change during growth of the seedlings. In the uppermost 0.5-mm region of the region in which the Casparian strip could be detected by fluorescence microscopy, the plasma membrane was not firmly attached to the cell wall. The development of the Casparian strip continued for about 42 h after dark-grown seedlings were transferred to the light, indicating that (i) the cells that have been determined to form the Casparian strip in darkness form the strip in the light, and that (ii) it takes about 42 h for the cells to complete formation of the strip. Cells in the hook of dark-grown seedlings did not form a Casparian strip when such seedlings were transferred to the light. The Casparian strip was formed in rapidly elongating internodes of light-grown seedlings when the seedlings were transferred to darkness. Light did not control the formation of the Casparian strip in roots.Abbreviation PBS phosphate-buffered saline     

Observations on the marginal ruffles of an established fibroblast-like cell line

Summary LW13K2 cells, a clone of a spontaneously in vitro transformed derivative of embryonic Lewis rat fibroblastic cells, were studied by phase contrast cine-light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The ruffles found at the advancing edge of cells grown on glass substrates in vitro form and recede in a period of less than one min if they do not make an attachment of the substrate. If they fail to make an attachment they may form pinocytotic channels near the leading edge as described by Price (1972) and/or collapse, generally backwards, towards the cell body. The spines which appear to reinforce the membranous ruffles are the last structures to disappear, and accumulate in an irregular array behind the ruffling edge; this area is behind that in which pinocytosis occurs. In comparison with the sparse numbers of ribosomes found in the trailing edge, they are present in notable concentrations near the leading, ruffling edge of the cell. No membrane vesicles have been found in or near the ruffling edges at the ruffle-spine concentration zone.     

Two types of luliberin-immunoreactive perikarya in the preoptic area of the rat

Summary At the light microscopic level, following immunostaining with a single antiserum against luliberin (LRF), two types of hormone-producing perikarya in the preoptic area are demonstrated. The two cell types differ in their morphological features: a bipolar, smooth-contoured cell type can be differentiated from an irregularly contoured unipolar type. Intermediate forms between both cell types occurring in the same area are not observed. Electron microscopically, both cell types contain labeled granules of similar size and immunoreactivity. It is dicussed whether the uneven surface of the one cell type is due to areas of synaptic contacts, and whether both cell types are integrated in different neuronal and functional circuits. Moreover, at the ultrastructural level, from the irregularly contoured LRF-producing perikarya a further positively stained cell type, probably a glial cell, can be differentiated. The specific labeling of the latter is caused by its content of immunoreactive lysosomal bodies. Differentiation between the labeled glial cells and the irregularly contoured LRF-producing perikarya is not possible at the light microscopic level.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/3) and by the Stiftung Volkswagenwerk     

The relationship between the size of mitochondria and the intensity of light that they scatter in different energetic states

The intensity of light scattered at 90° to the incident beam and the effective hydrodynamic radii of mitochondria incubated under a variety of conditions have been measured. Addition of high concentrations of uncouplers to respiring mitochondria resulted in a decrease in scatter which was not due to swelling. Addition of valinomycin to mitochondria depleted of substrate in K⁺-free medium produced an increase in scatter that was not due to shrinking. It is concluded that changes in the intensity of scattered light are not reliable indices of changes of volume of mitochondria, and that changes in conformation with changes in metabolic state dominate changes in light scatter. A molecular mechanism for the effect of metabolic state upon the scattered intensity is suggested.