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The developmental inhibition at the tight aggregate stage could be mediated through direct cell-to-cell contact, or by secretion of an inhibitory compound.
D. caveatum secretes one or several small compound(s) constitutively, even in the absence of prey cells, that induce a reversible block in prey cell development at the tight aggregate stage.
Our results suggest that, at a ratio of 1/103, D. caveatum induces a block in the development of D. discoideum at the tight aggregate stage: the collective rotational motion and morphogenesis of the tip are inhibited.
In this case, after 16 18 hours of development, it is likely that about 10% of the aggregate would be highly phagocytic D. caveatum and the D. discoideum amoebae may be so damaged that they cannot progress to the tight aggregate stage.
Under these conditions, development proceeded until the tight aggregate stage.
A hidden triplicate of the same chimera (Chim4, Chim6, and Chim12) appears also as a tight aggregate on the MDS plot (red ellipse named "1").
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Thus, it is not aggregation per se that represents a problem, but rather the fact that tight aggregates cannot be dispersed without damaging the cells.
In order to create large and tight aggregates a dual-polymer system was used using unmodified and sonicated flocculant.
In contrast, biofilms that grew in control media remained as tight aggregates for the duration of the experiments (72 hours).
To analyse cell-cell adhesion during development [29] cells were starved on phosphate agar plates and harvested after 10 to 12 hours once they had formed tight aggregates.
The activity of this promoter could be still observed in cells located at the base of tight aggregates (Fig. 2C) but was not detectable at later developmental stages, such as slug structures (Fig. 2D) or in spores (Fig. 2E).
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