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Nanobodies (Nbs) have great potential in immunoassays due to their exceptional physicochemical properties. With the immortal nature of Nbs and the ability to manipulate their structures using protein engineering, it will become increasingly valuable to understand what structural features of Nbs drive high stability, affinity, and selectivity. Here, we employed an anti-quinalphos Nb as a model to illustrate the structural basis of Nbs' distinctive physicochemical properties and the recognition mechanism. The results indicated that the Nb-11A-ligand complexes exhibit a "tunnel" binding mode formed by CDR1, CDR2, and FR3. The orientation and hydrophobicity of small ligands are the primary determinants of their diverse affinities to Nb-11A. In addition, the primary factors contributing to Nb-11A's limited stability at high temperatures and in organic solvents are the rearrangement of the hydrogen bonding network and the enlargement of the binding cavity. Importantly, Ala 97 and Ala 34 at the active cavity's bottom and Arg 29 and Leu 73 at its entrance play vital roles in hapten recognition, which were further confirmed by mutant Nb-F3. Thus, our findings contribute to a deeper understanding of the recognition and stability mechanisms of anti-hapten Nbs and shed new light on the rational design of novel haptens and directed evolution to produce high-performance antibodies.

The accurate analysis of chemical isomers plays an important role in the study of their different toxic effects and targeted detection of pollutant isomers in foods. The Alternaria mycotoxins tenuazonic acid (TeA) and iso-tenuazonic acid (ITeA) are two isomer mycotoxins with the lack of single analysis methods due to the similar structures. Antibody-based immunoassays exhibit high sensitivity and superior application in isomer-specific determination. Previously, various kinds of antibodies for TeA have been prepared in our group. Herein, highly specific nanobodies (Nbs) against ITeA mycotoxin were selected from immune nanobody phage display library, and one of Nbs, namely Nb(B3G3) exhibited excellent affinity, thermal stability as well as organic solvent tolerance. By molecular simulation and docking technology, it was found that stronger interaction between Nb(B3G3) and ITeA lead to higher affinity than that for its isomer TeA. Furthermore, a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) was established with a limit of detection (LOD) of 0.09 ng/mL for ITeA mycotoxin. The recovery rate of ITeA in spiked samples was analyzed with 84.8%-89.5% for rice, 78.3%-96.3% for flour, and 79.5%-90.7% for bread. A conventional LC-MS/MS method was used to evaluate the accuracy of this proposed icELISA, which showed a satisfactory consistent correlation. Since the convenient strategy for nanobody generation by phage display technology, this study provide new biorecognition elements and sensitive immunoassay for analysis of ITeA in foods.

Highlight: [•] We successfully yield the core-shelled SiO2–Ag nanospheres. [•] Proof the Ag shell's size effect the SPR conditions. [•] Enhancing the degradation efficiency is controlled by Ag nanoparticles’ size. [•] We replaced the formaldehyde and ammonia with glucose as the reduction in seed-mediated process. [ABSTRACT FROM AUTHOR]

The growth of roots towards aboveground litter layer is a common phenomenon in forest ecosystems. It is of great significance to examine the effects of root presence on litter decomposition for understanding nutrient cycling in forest ecosystems. We explored the effects of root growth on leaf litter decomposition, nutrient release and enzyme activities by establishing treatments with and without root with a one year field decomposition experiment in Phoebe zhennan and Castanopsis kawada-mii forests at Sanming, Fujian. The results showed that after 360 days decomposition, leaf litter mass remaining ratio in the treatment with root was 8.4% and 19.7% lower than control, respectively. The presence of root exhibited significant effect on litter decomposition during the 90-180 days. Compared with the control, the remaining ratio of leaf litter carbon, nitrogen and phosphorus were 6.0%, 19.1% and 20.6% lower in the treatment with root in the P. zhennan forest, and were 21.3%, 23.2% and 20.5% lower in the C. kawadamii forest, respectively. During the whole decomposition process, root presence did not affect the hydrolytic enzyme activity. After 180 days decomposition, the peroxidase activities in the treatment with root were 111.4% and 92.4% higher than control in the P. zhennan and C. kawadamii forests, respectively. The remaining ratio of leaf litter carbon, nitrogen and phosphorus were negatively correlated with the activities of cellobiohydrolase, β-glucosidase, acid phosphatase, and peroxidase. Root presence in litter layer could accelerate litter decomposition and nutrient release through nutrient uptake and stimulation of oxidase activity.

Objectives: Following publication of detailed national neonatal palliative care guidance, practical regional guidance, in the form of multidisciplinary 'checklists', was implemented aiming to improve the quality of neonatal palliative care., Methods: Case note audit was used to examine the quality of locally delivered neonatal palliative care before and after regional guidance implementation., Results: 27 patients were allocated to the 'before' cohort and 10 to the 'after' cohort. Introduction of the checklists was apparently associated with improvements in domains of pain relief and comfort care, monitoring, fluids and nutrition, completion of diagnostics, treatment ceiling decisions, resuscitation status and discussion with parents. Other support for parents was poorly adhered to., Conclusion: Regional guidance improved some aspects of palliative care delivery though other areas remained suboptimal. Other strategies, for example, consultation with paediatric palliative care services, need to be considered to further improve the quality of palliative care delivered to babies with life-limiting illnesses., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

We determined the water use efficiency and nitrogen and phosphorus concentrations of plants at different altitudes (600, 900, 1300, 1500, 1800, 2000, 2100 m) in Wuyi Mountains to understand the relationship of water use efficiency with foliar nutrients. The results showed that plant water use efficiency increased with altitude, and the leaf δ 18 O of tree showed no significant variance with altitude. On the whole, leaf nitrogen concentration showed no obvious trend, while leaf phosphorus concentration at high altitude was significantly higher than that at low altitude. No significant relationship between water use efficiency and foliar nitrogen concentration was found in this study, but water use efficiency had a positive correlation with foliar phosphorus concentration. In conclusion, the change of water use efficiency was mainly caused by the difference in photosynthetic rate. The effect of water status on plant water use efficiency was not significant. The variances of leaf phosphorus concentrations along the altitudinal gradient may affect photosynthetic rate and in turn the water use efficiency of plant in this area.