Auxin and nitric oxide control indeterminate nodule formation

Background  

Rhizobia symbionts elicit root nodule formation in leguminous plants. Nodule development requires local accumulation of auxin. Both plants and rhizobia synthesise auxin. We have addressed the effects of bacterial auxin (IAA) on nodulation by using Sinorhizobium meliloti and Rhizobium leguminosarum bacteria genetically engineered for increased auxin synthesis.     

Photochemistry and DNA-affinity of some pyrimidine-substituted styryl-azinium iodides

The relaxation properties of the excited states of three iodides of trans-1,2-diarylethene analogues (where one aryl group is a methylpyridinium, methylquinolinium or dimethylimidazolium group and the other one is a phenyl ring para-substituted by a pyrimidine ring) have been investigated in buffered (pH = 7) aqueous solution. As found in previous works for several analogues, these quaternized salts undergo efficient trans→cis photoisomerization while the yield of the radiative deactivation is very small at room temperature. The solvent effect on the spectral behaviour indicates the occurrence of intramolecular charge transfer which can induce interesting non-linear optical properties. The results of a study of the interactions of these salts with DNA, which might affect the cell metabolism, showed a relatively modest binding affinity for the pyridinium and imidazolium salts and a more substantial affinity for the quinolinium analogue. The formation of ligand-DNA complexes affects only slightly the radiative relaxation yield while leading to a relevant reduction of the isomerization yield. Measurements of the linear dichroism behaviour of the three compounds and comparison with three analogues bearing furan or thienyl groups, which have been found to display different affinity with DNA in previous works, gave interesting information on the nature of the ligand-DNA binding of these compounds.     

Inter-limb coordination, strength, jump, and sprint performances following a youth men's basketball game

This study aimed to verify whether basketball players are able to maintain strength (handgrip), jump (countermovement jump [CMJ]), sprint (10 m and 10 m bouncing the ball [10 mBB]), and interlimb coordination (i.e., synchronized hand and foot flexions and extensions at 80, 120, and 180 bpm) performances at the end of their game. Ten young (age 15.7 ± 0.2 years) male basketball players volunteered for this study. During the friendly game, heart rate (HR), rate of perceived exertion (RPE), and rate of muscle pain (RMP) were assessed to evaluate the exercise intensity. Overall, players spent 80% of the time playing at intensities higher than 85% HRmax. Main effects (p < 0.05) for game periods emerged for HR and the number of players involved in a single action, with lower occurrence of maximal efforts and higher involvement of teammates after the first 2 periods. At the end of the game, players reported high (p < 0.05) RPE (15.7 ± 2.4) and RMP (5.2 ± 2.3) values; decreased (p < 0.05) sprint capabilities (10 m: pre = 1.79 ± 0.09 seconds, post = 1.84 ± 0.08 seconds; 10 mBB: pre = 1.81 ± 0.11 seconds, post = 1.96 ± 0.08 seconds); increased (p < 0.05) interlimb coordination at 180 bpm (pre = 33.3 ± 20.2 seconds, post = 43.9 ± 19.8 seconds); and maintained jump (pre = 35.2 ± 5.2 cm, post = 35.7 ± 5.2 cm), handgrip (pre = 437 ± 73 N, post = 427 ± 55 N), and coordinative performances at lower frequencies of executions (80 bpm: pre = 59.7 ± 1.3 seconds, post = 60.0 ± 0.0 seconds; 120 bpm: pre = 54.7 ± 12.3 seconds, post = 57.3 ± 6.7 seconds). These findings indicate that the heavy load of the game exerts beneficial effects on the efficiency of executive and attentive control functions involved in complex motor behaviors. Coaches should structure training sessions that couple intense physical exercises with complex coordination tasks to improve the attentional capabilities of the players.     

Mechanism of inhibition of enveloped virus membrane fusion by the antiviral drug arbidol

The broad-spectrum antiviral arbidol (Arb) inhibits cell entry of enveloped viruses by blocking viral fusion with host cell membrane. To better understand Arb mechanism of action, we investigated its interactions with phospholipids and membrane peptides. We demonstrate that Arb associates with phospholipids in the micromolar range. NMR reveals that Arb interacts with the polar head-group of phospholipid at the membrane interface. Fluorescence studies of interactions between Arb and either tryptophan derivatives or membrane peptides reconstituted into liposomes show that Arb interacts with tryptophan in the micromolar range. Interestingly, apparent binding affinities between lipids and tryptophan residues are comparable with those of Arb IC50 of the hepatitis C virus (HCV) membrane fusion. Since tryptophan residues of membrane proteins are known to bind preferentially at the membrane interface, these data suggest that Arb could increase the strength of virus glycoprotein's interactions with the membrane, due to a dual binding mode involving aromatic residues and phospholipids. The resulting complexation would inhibit the expected viral glycoprotein conformational changes required during the fusion process. Our findings pave the way towards the design of new drugs exhibiting Arb-like interfacial membrane binding properties to inhibit early steps of virus entry, i.e., attractive targets to combat viral infection.     

FLO11-based model for air-liquid interfacial biofilm formation by Saccharomyces cerevisiae

Sardinian wine strains of Saccharomyces cerevisiae used to make sherry-like wines form a biofilm at the air-liquid interface at the end of ethanolic fermentation, when grape sugar is depleted and further growth becomes dependent on access to oxygen. Here, we show that FLO11, which encodes a hydrophobic cell wall glycoprotein, is required for the air-liquid interfacial biofilm and that biofilm cells have a buoyant density greater than the suspending medium. We propose a model for biofilm formation based on an increase in cell surface hydrophobicity occurring at the diauxic shift. This increase leads to formation of multicellular aggregates that effectively entrap carbon dioxide, providing buoyancy. A visible biofilm appears when a sufficient number of hydrophobic cell aggregates are carried to and grow on the liquid surface.     

Evolution of an acylase active on cephalosporin C

Semisynthetic cephalosporins are synthesized from 7-amino cephalosporanic acid, which is produced by chemical deacylation or by a two-step enzymatic process of the natural antibiotic cephalosporin C. The known acylases take glutaryl-7-amino cephalosporanic acid as a primary substrate, and their specificity and activity are too low for cephalosporin C. Starting from a known glutaryl-7-amino cephalosporanic acid acylase as the protein scaffold, an acylase gene optimized for expression in Escherichia coli and for molecular biology manipulations was designed. Subsequently we used error-prone PCR mutagenesis, a molecular modeling approach combined with site-saturation mutagenesis, and site-directed mutagenesis to produce enzymes with a cephalosporin C/glutaryl-7-amino cephalosporanic acid catalytic efficiency that was increased up to 100-fold, and with a significant and higher maximal activity on cephalosporin C as compared to glutaryl-7-amino cephalosporanic acid (e.g., 3.8 vs. 2.7 U/mg protein, respectively, for the A215Y-H296S-H309S mutant). Our data in a bioreactor indicate an ~90% conversion of cephalosporin C to 7-amino-cephalosporanic acid in a single deacylation step. The evolved acylase variants we produced are enzymes with a new substrate specificity, not found in nature, and represent a hallmark for industrial production of 7-amino cephalosporanic acid.     

A bunch-oligonucleotide forming stable monomolecular quadruplex containing a T-tetrad

The chemical synthesis of bunch-ODN I and II prone to form quadruplex structures containing G-and T-tetrads has been reported. Structural studies were performed by 1H-NMR and CD melting experiments.     

Synthesis and characterization of monomolecular DNA G-quadruplexes formed by tetra-end-linked oligonucleotides

Guanine-rich DNA sequences are widely dispersed in the eukaryotic genome and are abundant in regions with relevant biological significance. They can form quadruplex structures stabilized by guanine quartets. These structures differ for number and strand polarity, loop composition, and conformation. We report here the syntheses and the structural studies of a set of interconnected d(TG(4)T) fragments which are tethered, with different orientations, to a tetra-end-linker in an attempt to force the formation of specific four-stranded DNA quadruplex structures. Two synthetic strategies have been used to obtain oligodeoxyribonucleotide (ODN) strands linked with their 3'- or 5'-ends to each of the four arms of the linker. The first approach allowed the synthesis of tetra-end-linked ODN (TEL-ODN) containing the four ODN strands with a parallel orientation, while the latter synthetic pathway led to the synthesis of TEL-ODNs each containing antiparallel ODN pairs. The influence of the linker at 3'- or 5'-ODN, on the quadruplex typology and stability, in the presence of sodium or potassium ions, has been investigated by circular dichroism (CD), CD thermal denaturation, (1)H NMR experiments at variable temperature, and molecular modeling. All synthesized TEL-ODNs formed parallel G-quadruplex structures. Particularly, the TEL-ODN containing all parallel ODN tracts formed very stable parallel G-quadruplex complexes, whereas the TEL-ODNs containing antiparallel ODN pairs led to relatively less stable parallel G-quadruplexes. The molecular modeling data suggested that the above antiparallel TEL-ODNs can adopt parallel G-quadruplex structures thanks to a considerable folding of the tetra-end-linker around the whole quadruplex scaffold.