Cu-II-directed self-assembly of fullerenols to ameliorate copper stress in maize seedlings.

Widespread use of copper-based agrochemical may cause copper excessive accumulation in agricultural soil to seriously threaten crop production. Recently, fullerenols are playing important roles in helping crops build resistance to abiotic stresses by giving ingenious and successful resolutions. However, there is a lack of knowledge on their beneficial effects in crops under stresses induced by heavy metals. Herein, the visual observation of Cu2+-mediated assembly of fullerenols via electrostatic and coordination actions was carried out in vitro, showing that water-soluble nanocomplexes and water-insoluble cross-linking nanohybrids were selectively fabricated by precisely adjusting feeding ratios of fullerenols and CuSO4. Furthermore, maize simultaneous exposure of fullerenols and CuSO4 solutions was tested to investigate the comparative effects of seed germination and seedling growth relative to exposure of CuSO4 alone. Under moderate Cu2+ stresses (40 and 80 µM), fullerenols significantly mitigated the detrimental effects of seedlings, including phenotype, root and shoot elongation, biomass accumulation, antioxidant capacity, and Cu2+ uptake and copper transporter-related gene expressions in roots. Under 160 µM of Cu2+ as a stressor, fullerenols also accelerated germination of Cu2+-stressed seeds eventually up to the level of the control. Summarily, fullerenols can enhance tolerance of Cu2+-stressed maize mainly due to direct detoxification through fullerenol-Cu2+ interactions restraining the Cu2+ intake into roots and reducing free Cu2+ content in vivo, as well as fullerenol-maize interactions to enhance resistance by maintaining balance of reactive oxygen species and optimizing the excretion and transport of Cu2+. This will unveil valuable insights into the beneficial roles of fullerenols and its mechanism mode in alleviating heavy metal stress on crop plants.

Directed assembly of fullerenols via electrostatic and coordination interactions to fabricate diverse and water-soluble metal cation-fullerene nanocluster complexes.

Metal ion-nanocluster coordination complexes can produce a variety of functional engineered nanomaterials with promising characteristics to enable widespread applications. Herein, the visualization observation of the interactions of metal ions and fullerene derivatives, particularly anionic fullerenols (Fol), were carried out in aqueous solutions. The alkali metal salts only resulted in salting out of Fol to gain re-soluble sediments, whereas multivalent metal cations (Mn+, n = 2, 3) modulated further assembly of Fol to produce insoluble hybrids. These provide crucial insights into the directed assembly of Fol that two major forces involved in actuation are electrostatic and coordination effects. Through the precise modulation of feed ratios of Fol to Mn+, a variety of water-soluble Mn+@Fol coordination complexes were facilely prepared and subsequently characterized by various measurements. Among them, X-ray photoelectron spectra validated the coordination effects through the metal cation and oxygen binding feature. Transmission electron microscopy delivered valuable information about diverse morphologies and locally-ordered microstructures at the nanoscale. This study opens a new opportunity for developing a preparation strategy to fabricate water-soluble metal cation-fullerenol coordination complexes with various merits for potential application in biomedical fields.

Cyclic di-GMP inhibits nitrate assimilation by impairing the antitermination function of NasT in Pseudomonas putida.

The ubiquitous bacterial second messenger cyclic diguanylate (c-di-GMP) coordinates diverse cellular processes through its downstream receptors. However, whether c-di-GMP participates in regulating nitrate assimilation is unclear. Here, we found that NasT, an antiterminator involved in nitrate assimilation in Pseudomonas putida, specifically bound c-di-GMP. NasT was essential for expressing the nirBD operon encoding nitrite reductase during nitrate assimilation. High-level c-di-GMP inhibited the binding of NasT to the leading RNA of nirBD operon (NalA), thus attenuating the antitermination function of NasT, resulting in decreased nirBD expression and nitrite reductase activity, which in turn led to increased nitrite accumulation in cells and its export. Molecular docking and point mutation assays revealed five residues in NasT (R70, Q72, D123, K127 and R140) involved in c-di-GMP-binding, of which R140 was essential for both c-di-GMP-binding and NalA-binding. Three diguanylate cyclases (c-di-GMP synthetases) were found to interact with NasT and inhibited nirBD expression, including WspR, PP_2557, and PP_4405. Besides, the c-di-GMP-binding ability of NasT was conserved in the other three representative Pseudomonas species, including P. aeruginosa, P. fluorescens and P. syringae. Our findings provide new insights into nitrate assimilation regulation by revealing the mechanism by which c-di-GMP inhibits nitrate assimilation via NasT.

Second Messenger c-di-GMP Modulates Exopolysaccharide Pea-Dependent Phenotypes via Regulation of eppA Expression in Pseudomonas putida.

The exopolysaccharide (EPS) Pea is essential for wrinkly colony morphology, pellicle formation, and robust biofilm production in Pseudomonas putida. The second messenger cyclic diguanylate monophosphate (c-di-GMP) induces wrinkly colony morphology in P. putida through an unknown mechanism(s). Herein, we found that c-di-GMP modulates wrinkly colony morphology via the regulation of expression of eppA (PP_5586), a small individually transcribed gene of 177 bp, and this gene was adjacent to the upstream region of the pea cluster. Phenotype observation revealed that eppA was essential for Pea-dependent phenotypes. The deletion of eppA led to a smooth colony morphology and impaired biofilm, which was analogous to the phenotypes with loss of the entire pea operon. eppA expression was positively regulated by c-di-GMP via the transcriptional effector FleQ, and eppA was essential for the c-di-GMP-induced wrinkly colony morphology. Structure prediction results implied that EppA had two transmembrane regions, and Western blotting revealed that EppA was located on the cell membrane. Transcriptomic analysis indicated that EppA had no significant effect on the transcriptomic profile of P. putida. A bacterial two-hybrid (BTH) assay suggested that there was no direct interaction between EppA and the proteins in the pea cluster and adjacent operons. Overall, these findings reveal that EppA is essential for Pea-dependent phenotypes and that c-di-GMP modulates Pea-dependent phenotypes via regulation of eppA expression in P. putida. IMPORTANCE Microbe-secreted EPSs are high-molecular-weight polysaccharides that have the potential to be used as industrially important biomaterials. The EPS Pea in P. putida is essential for wrinkly colony morphology and pellicle formation. Here, we identified a function-unknown protein, EppA, which was also essential for Pea-dependent wrinkly colony morphology and pellicle formation, and EppA was probably involved in Pea secretion. Meanwhile, our results indicated that the second messenger c-di-GMP positively regulated the expression of EppA, resulting in Pea-dependent wrinkly colony morphology. Our results reveal the relationship of c-di-GMP, EppA, and Pea-dependent phenotypes and provide a possible pathway to construct genetically engineered strains for high Pea production.

The two-component system TarR-TarS is regulated by c-di-GMP/FleQ and FliA and modulates antibiotic susceptibility in Pseudomonas putida.

Two-component systems (TCSs) are predominant means by which bacteria sense and respond to environment signals. Genome of Pseudomonas putida contains dozens of putative TCS-encoding genes, but phenotypical-genotypical correlation and transcriptional regulation of these genes are largely unknown. Herein, we characterized function and transcriptional regulation of a conserved P. putida TCS, named TarR-TarS. TarS (PP_0769) encodes a potential histidine kinase, and tarR (PP_0768) encodes a potential response regulator. Protein-protein interaction assay and phosphorylation assay confirmed that TarR-TarS was a functional TCS. Growth assay under antibiotics revealed that TarR-TarS positively regulated bacterial resistance to multiple antibiotics. Pull-down assay revealed that TarR directly interacted with PP_0800 (a hypothetical protein) and GroEL (the chaperonin). GroEL played a positive role in antibiotic resistance, while PP_0800 seemed to have no effect on antibiotic resistance. The regulator FleQ indirectly activated tarR-tarS transcription. However, the second messenger c-di-GMP antagonized FleQ activation to inhibit tarR-tarS transcription. The sigma factor FliA directly activated tarR-tarS transcription via a consensus motif. These findings reveal function and transcriptional regulation of TarR-TarS, and enrich knowledge regarding the relationship between c-di-GMP and antibiotic susceptibility in P. putida.

The anterior versus posterior approach for the treatment of ossification of the posterior longitudinal ligament in the cervical spine: A systematic review and meta-analysis.

Study Design: Systematic review and meta-analysis.Objective: To compare the effectiveness and safety between anterior and posterior approach, and determine the best surgical methods for the treatment of ossification of the posterior longitudinal ligament (OPLL) in the cervical spine.Methods: We searched the Cochrane Library, PubMed, CNKI and Wanfang Med Data databases from January 2007 to March 2018. Japanese Orthopaedic Association (JOA) scores, cervical lordosis, functional recovery rates, excellent and good outcomes of the surgical approaches, and complication and reoperation rates were analyzed. RevMan 5.3 was utilized for data analysis.Results: Eleven studies were included in the meta-analysis. By comparing the anterior and posterior approaches for the treatment of OPLL in the cervical spine, statistically significant differences were found in the preoperative initial JOA, the postoperative final JOA scores, functional recovery rates, complication rates, excellent and good outcomes of the surgical approaches and reoperation rates. However, no statistically significant difference in the occurrence of the preoperative and postoperative cervical lordosis was noted.Conclusion: The anterior approach is superior to the posterior approach in terms of the postoperative final JOA score, functional recovery rate, and clinical outcomes. Although the complication and reoperation rates of the anterior approach are higher than those of the posterior approach. We recommend the anterior approach for the treatment of OPLL when patients with occupying ratio ≥ 60%. In addition, high-quality studies with long-term follow-up and large sample size are also needed.

Identification and epistasis analysis of quantitative trait loci for zeaxanthin concentration in maize kernel across different generations and environments.

Zeaxanthin, a natural fat-soluble pigment, not only increases plant resistance, but also has vital significance for human health. However, quantitative trait loci (QTL) and the epistatic effects of zeaxanthin concentration in maize kernel have not been well studied. To identify QTLs and analyse the epistatic effects of zeaxanthin concentration in maize kernel, two sets of segregating generations derived from the cross between HuangC (a high zeaxanthin concentration inbred line) and Rezi1 (a low zeaxanthin concentration inbred line) were evaluated in three different environments. One major-effect QTL, qZea6a, explains 41.4-71.4% of the phenotypic variation and two QTLs, qZea4a and qZea3a, show LOD > 3 for zeaxanthin concentration detected over two generations and three different environments. Four of the ten QTL pairs show epistatic effects, explaining 7.34-14.3% of the phenotypic variance. Furthermore, additivity was the major allelic action at zeaxanthin concentration QTLs located in F2 and F2:3 populations and plants with homozygous HuangC alleles have a strong genetic ability in enhancing zeaxanthin concentration in maize kernel. These results will contribute to understanding these complex loci better and provide awareness about zeaxanthin concentration to maize breeders and scientists involved in maize research.

Bacterial Infection in Deep Paraspinal Muscles in a Parturient Following Epidural Analgesia: A Case Report and Literature Review: A CARE-Compliant Article.

Bacterial infection related to epidural catheterizations could occur. In general, the incidence of postoperative infection at the insertion site is very low. Paucity literatures are reported for paraspinal muscle infection after epidural analgesia in parturient. We report a case of paraspinal muscle infection shortly after epidural analgesia in a parturient, who was subjected to because of threatened preterm labor. Epidural morphine was administered for 2 days for childbirth pain control. She began to have constant low-back pain and fever on postpartum Day 2. Magnetic resonance image revealed a broad area of subcutaneous edema with a continuum along the catheter trajectory deep to the paraspinal muscles. A catheter-related bacterial infection was suspected. The surgical debridement and drainage was required combined with intravenous antibiotics on postpartum Day 3. She was soon cured uncomplicatedly. Epidural analgesia is effective to control labor pain and, in general, it is safe. However, the sequelae of complicated infection may be underestimated. A literature search yielded 7 other cases of catheter-related epidural abscess or soft tissue infection. Vigilance for these infections, especially in postpartum patients with backache, is needed. Moreover, early detection and proper treatment of infectious signs at postanesthetic visit are very important.