794 research outputs found
FlhF, a signal recognition particle-like GTPase, is involved in the regulation of flagellar arrangement, motility behaviour and protein secretion in Bacillus cereus
Flagellar arrangement is a highly conserved feature within bacterial species. However, only a few genes regulating cell flagellation have been described in polar flagellate bacteria. This report demonstrates that the arrangement of flagella in the peritrichous flagellate Bacillus cereus is controlled by flhF. Disruption of flhF in B. cereus led to a reduction in the number of flagella from 10-12 to 1-3 filaments per cell in the insertion mutant MP06. Moreover, compared to the parental strain, MP06 exhibited: (i) shorter smooth swimming phases, causing reduced swimming motility but not affecting chemotaxis; (ii) complete inhibition of swarming motility, as differentiated swarm cells were never detected; (iii) an increased amount of extracellular proteins; and (iv) differential export of virulence determinants, such as haemolysin BL (HBL), phosphatidylcholine-preferring phospholipase C (PC-PLC) and non-haemolytic enterotoxin (NHE). Introduction of a plasmid harbouring flhF (pDGflhF) into MP06 completely restored the wild-type phenotype in the trans-complemented strain MP07. B. cereus flhF was found to constitute a monocistronic transcriptional unit and its overexpression did not produce abnormal features in the wild-type background. Characterization of a B. cereus mutant (MP05) carrying a partial flhF deletion indicated that the last C-terminal domain of FlhF is involved in protein export while not required for flagellar arrangement and motility behaviour. Taken together, these data suggest that B. cereus FlhF is a promising candidate for connecting diverse cellular functions, such as flagellar arrangement, motility behaviour, pattern of protein secretion and virulence phenotype
INVOLVEMENT OF BASIC FIBROBLAST GROWTH-FACTOR IN SURAMIN-INDUCED INHIBITION OF V79/AP4 FIBROBLAST CELL-PROLIFERATION
The V79/AP4 Chinese hamster fibroblasts were densely stained with the anti-basic fibroblast growth factor (bFGF) antibody demonstrating an endogenous production of the peptide. The in vitro proliferation of these cells was stimulated by exogenous bFGF and the maximum growth (259% increase in H-3-thymidine incorporation into DNA) was reached with bFGF 10 ng ml-1. Inhibition of bFGF-mediated mitogenic pathway was obtained with a 15-mer antisense oligodeoxynucleotide targeted against bFGF mRNA and with suramin, a drug which blocks the biological activity of heparin-binding growth factors. bFGF antisense oligomer reduced the synthesis of DNA by 79.5 and 89.5% at 20 and 60 muM, respectively; this effect was reversed by the addition of exogenous bFGF to the culture medium. A short-term exposure to suramin 300 mug ml-1 produced a modest reduction in H-3-thymidine incorporation but suppressed the mitogenic effect of bFGF on V79/AP4 cells. In cells treated with suramin 300 mug ml-1 the drug concentration increased linearly over 3 days, reaching 13.15 mug mg-1 of protein; cell proliferation was inhibited in a dose-related manner as evaluated by the colony formation assay (IC50: 344.22 mug ml-1) and by the number of mitoses observed in culture. Furthermore, the drug induced ultrastructural alterations, consisting of perinuclear cisternae swelling, chromatin condensation, nucleolar segregation and cytoplasmic vacuolations. These findings demonstrated that the endogenous production of bFGF plays an important role in V79/AP4 fibroblasts proliferation, and the inhibition of bFGF-mediated mitogenic signalling with bFGF antisense oligomer or suramin is an effective mean of reducing cell growth
Involvement of basic fibroblast growth factor in suramin-induced inhibition of V79/AP4 fibroblast cell proliferation.
The V79/AP4 Chinese hamster fibroblasts were densely stained with the anti-basic fibroblast growth factor (bFGF) antibody demonstrating an endogenous production of the peptide. The in vitro proliferation of these cells was stimulated by exogenous bFGF and the maximum growth (259% increase in 3H-thymidine incorporation into DNA) was reached with bFGF 10 ng ml-1. Inhibition of bFGF-mediated mitogenic pathway was obtained with a 15-mer antisense oligodeoxynucleotide targeted against bFGF mRNA and with suramin, a drug which blocks the biological activity of heparin-binding growth factors. bFGF antisense oligomer reduced the synthesis of DNA by 79.5 and 89.5% at 20 and 60 microM, respectively; this effect was reversed by the addition of exogenous bFGF to the culture medium. A short-term exposure to suramin 300 micrograms ml-1 produced a modest reduction in 3H-thymidine incorporation but suppressed the mitogenic effect of bFGF on V79/AP4 cells. In cells treated with suramin 300 micrograms ml-1 the drug concentration increased linearly over 3 days, reaching 13.15 micrograms mg-1 of protein; cell proliferation was inhibited in a dose-related manner as evaluated by the colony formation assay (IC50: 344.22 micrograms ml-1) and by the number of mitoses observed in culture. Furthermore, the drug induced ultrastructural alterations, consisting of perinuclear cisternae swelling, chromatin condensation, nucleolar segregation and cytoplasmic vacuolations. These findings demonstrated that the endogenous production of bFGF plays an important role in V79/AP4 fibroblasts proliferation, and the inhibition of bFGF-mediated mitogenic signalling with bFGF antisense oligomer or suramin is an effective mean of reducing cell growth
Plastic changes in the spinal cord in motor neuron disease.
In the present paper, we analyze the cell number within lamina X at the end stage of disease in a G93A mouse model of ALS; the
effects induced by lithium; the stem-cell like phenotype of lamina X cells during ALS; the differentiation of these cells towards
either a glial or neuronal phenotype. In summary we found that G93A mouse model of ALS produces an increase in lamina
X cells which is further augmented by lithium administration. In the absence of lithium these nestin positive stem-like cells
preferentially differentiate into glia (GFAP positive), while in the presence of lithium these cells differentiate towards a neuronlike
phenotype (III-tubulin, NeuN, and calbindin-D28K positive). These effects of lithium are observed concomitantly with
attenuation in disease progression and are reminiscent of neurogenetic effects induced by lithiumin the subependymal ventricular
zone of the hippocampus
Gaseous argon time projection chamber with electroluminescence enhanced optical readout
Systematic uncertainties in accelerator oscillation neutrino experiments
arise mostly from nuclear models describing neutrino-nucleus interactions. To
mitigate these uncertainties, we can study neutrino-nuclei interactions with
detectors possessing enhanced hadron detection capabilities at energies below
the nuclear Fermi level. Gaseous detectors not only lower the particle
detection threshold but also enable the investigation of nuclear effects on
various nuclei by allowing for changes in the gas composition. This approach
provides valuable insights into the modelling of neutrino-nucleus interactions
and significantly reduces associated uncertainties. Here, we discuss the design
and first operation of a gaseous argon time projection chamber optically read.
The detector operates at atmospheric pressure and features a single stage of
electron amplification based on a thick GEM. Here, photons are produced with
wavelengths in the vacuum ultraviolet regime. In an optical detector the
primary constraint is the light yield. This study explores the possibility of
increasing the light yield by applying a low electric field downstream of the
ThGEM. In this region, called the electroluminescence gap, electrons propagate
and excite the argon atoms, leading to the subsequent emission of photons. This
process occurs without any further electron amplification, and it is
demonstrated that the total light yield increases up to three times by applying
moderate electric fields of the order of 3~kV/cm. Finally, an indirect method
is discussed for determining the photon yield per charge gain of a ThGEM,
giving a value of 18.3 photons detected per secondary electron
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