Isoenzymes of soluble starch synthetase from Oryza sativa grains

Two isoenzymic fractions of soluble ADP-glucose: α-1,4-glucan-4-glucosyltransferase were obtained from developing (non-waxy) rice grains by gradient elution through DEAE-cellulose. After Sephadex G-200 chromatography, fractions I and II were electrophoretically homogeneous and have MW values of 110000 and 69000, respectively. Sodium dodecyl sulfate gel electrophoresis of fraction I produced five bands with MW of 12000, 26000, 50000, 70000, and 105000 while fraction II gave two bands with MW of 12000 and 22000. Fraction II, which contains 1·7% carbohydrate, was active in the absence of added primer while fraction I, which does not contain carbohydrate, required primer.     

Serial sectioning study of some meiotic stages in Scaptericus borrelli (Grylloidea)

An electron microscope study of the spermatocytes I of Scaptericus borrelli was performed using serial sections and tridimensional reconstruction. The stages examined were: mid-pachytene, early diplotene and the so-called diffuse stage. At mid-pachytene the nucleolus (associated to an autosome) shows a glomerular array formed of supernumerary synaptonemal complexes (extra-complexes, ECs). At early diplotene the autosomes still show remnants of synaptonemal complexes (SC) and the sex chromosome is formed by two substances: one is homogeneous (chromatin-like) the other is granular and dense. A short number of ECs occur in the latter. — At the diffuse stage (lobulated in this paper) the SCs remnants and the ECs have disappeared and the granular material is assembled in a nuclear lobule. This lobule was found associated with one centriole. — A tridimensional model of a diplotene nucleus and models of diplotene bivalents are shown. — The capacity to form repetitive components of the synaptonemal complex type by spermatocytes of the super-Family Grylloidea is discussed.     

C6: A Monoclonal Antibody Specific for a Fibronectin Epitope Situated at the Interface between the Oncofoetal Extra-Domain B and the Repeat III8

Background

Fibronectin (FN) is a large multidomain molecule that is involved in many cellular processes. Different FN isoforms arise from alternative splicing of the pre-mRNA including, most notably, the FN isoform that contains the “extra-domain-B” (ED-B). The FN isoform containing ED-B (known as B-FN) is undetectable in healthy adult tissues but is present in large amounts in neoplastic and foetal tissues as well as on the blood vessels during angiogenesis. Thus, antibodies specific for B-FN can be useful for detecting and targeting neoplastic tissues in vivo. We previously characterised C6, a new monoclonal antibody specific for human B-FN and we suggested that it reacts with the B-C loop of the type III repeat 8 which is masked in FN isoforms lacking ED-B and that the insertion of ED-B in FN molecules unmasked it. Here we have now consolidated and refined the characterization of this B-FN specific antibody demonstrating that the epitope recognized by C6 also includes loop E-F of ED-B.

Methodology

We built the three dimensional model of the variable regions of the mAb C6 and of the FN fragment EDB-III8 and performed protein:protein docking simulation using the web server ClusPro2.0. To confirm the data obtained by protein:protein docking we generated mutant fragments of the recombinant FN fragment EDB-III8 and tested their reactivity with C6.

Conclusion

The monoclonal antibody C6 reacts with an epitope formed by the B-C loop of domain III8 and the E-F loop of ED-B. Both loops are required for an immunological reaction, thus this monoclonal is strictly specific for B-FN but the part of the epitope on III8 confers the human specificity.     

Can resistance against quorum-sensing interference be selected?

Quorum-sensing (QS) interference is a novel therapy to fight bacterial infections that, unlike conventional antibiotic treatments, is focused on reducing the damage caused by pathogens (virulence) rather than focused on inhibiting their growth. Given this ideal, it was predicted that this approach will be impervious to or at least much less prone to resistance in bacterial populations. However, recently, resistance mechanisms against well-characterized quorum quenchers (QQs) have been found in the laboratory as well as in clinical strains, demonstrating that the rise of resistance against these kinds of compounds is possible. Nevertheless, it has been argued that even if resistance mechanisms against QS interference exist, this fact does not guarantee that resistance will spread. In the present work, we discuss recent insights derived from the latest experiments to address this question. In addition, we explain how environmental conditions like the stress produced by the host immune system may influence the selection of resistance and eventually lead to the selection of QS interference-resistant bacteria in a clinical setting.     

Molecular mechanisms underlying the entrance in secondary dormancy of Arabidopsis seeds

As seasons change, dormant seeds cycle through dormant states until the environmental conditions are favourable for seedling establishment. Dormancy cycle is widespread in the plant kingdom allowing the seeds to display primary and secondary dormancy. Several reports in the last decade have focused on understanding the molecular mechanisms of primary dormancy, but our knowledge regarding secondary dormancy is limited. Here, we studied secondary dormancy induced in Arabidopsis thaliana by incubating seeds at 25 °C in darkness for 4 d. By physiological, pharmacological, expression and genetics approaches, we demonstrate that (1) the entrance in secondary dormancy involves changes in the content and sensitivity to GA, but the content and sensitivity to ABA do not change, albeit ABA is required; (2) RGL2 promotes the entrance in secondary dormancy through ABI5 action; and (3) multivariate analysis with 18 geographical and environmental parameters of accession collection place suggests that temperature is an important variable influencing the induction of secondary dormancy in nature.     

Ethylene Alleviates the Suppressive Effect of Phosphate on Arbuscular Mycorrhiza Formation

Phosphorus is one of the most important macronutrients required for plant growth. Plants have evolved many strategies for inorganic phosphorus (Pi) acquisition, including the symbiotic pathways, involving the formation of mycorrhiza. With regard to arbuscular mycorrhiza (AM), high Pi availability has long been known to negatively affect this association, although the underlying mechanism is unknown. In the present study, the interactive role played by ethylene and Pi in AM regulation was investigated in the tomato-Rhizophagus irregularis symbiosis. AM fungal colonization was analysed in epi, rin and NRO ethylene-responsive mutants under different Pi availability conditions, with a focus on the late stages of the interaction. Although Pi inhibited mycorrhizal parameters in the ethylene-insensitive rin mutant and wild-type cultivars, it did not alter the mycorrhization of the epi tomato mutant, which exhibits a constitutive ethylene-induced response. As with the colonization parameters, root ethylene content and the expression of AM-related and ethylene receptor 6 genes were inhibited by Pi in wild-type cultivars and rin mutants, but were unaffected or slightly activated in epi plants. The application of ethephon offsets the negative impact on the mycorrhizal development caused by the application of Pi. This compensation effect is dose dependent and was ineffective in the NRO mutant, which is more insensitive to the action of ethylene. Our results provide evidence that ethylene signalling negatively affects the suppressive effect of Pi on AM formation and suggests an overlap between this suppressive effect and the regulatory mechanism of Pi-starvation response pathway in plants mediated by ethylene.