Molecular cloning of the Lon protease gene from Thermus thermophilus HB8 and characterization of its gene product.

The gene encoding Lon protease was isolated from an extreme thermophile, Thermus thermophilus HB8. Sequence analysis demonstrated that the T. thermophilus Lon protease gene (TT-lon) contains a protein-coding sequence consisting of 2385 bp which is approximately 56% homologous to the Escherichia coli counterpart. As expected, the G/C content of TT-lon was 68%, which is significantly higher than that of the E. coli lon gene (52% G/C). The amino acid sequence of T. thermophilus Lon protease (TT-Lon) predicted from the nucleotide sequence contained several unique sequences conserved in other Lon proteases: (a) a cysteine residue at the position just before the putative ATP-binding domain; (b) motif A and B sequences required for composition of the ATP-binding domain; and (c) a serine residue at the proteolytic active site. Expression of TT-lon under the control of the T7 promoter in E. coli produced an 89-kDa protein with a yield of approximately 5 mg.L-1. Recombinant TT-Lon (rTT-Lon) was purified to homogeneity by sequential column chromatography. The peptidase activity of rTT-Lon was activated by ATP and alpha-casein. rTT-Lon cleaved succinyl-phenylalanyl-leucyl-phenylalanyl-methoxynaphthylamide much more efficiently than succinyl-alanyl-alanyl-phenylalanyl-methoxynaphthylamide, whereas both peptides were cleaved with comparable efficiencies by E. coli Lon. These results suggest that there is a difference between TT-Lon and E. coli Lon in substrate specificity. rTT-Lon most effectively cleaved substrate peptides at 70 degrees C, which was significantly higher than the optimal temperature (37 degrees C) for E. coli Lon. Together, these results indicate that the TT-lon gene isolated from T. thermophilus HB8 actually encodes an ATP-dependent thermostable protease Lon.     

OsCESA9 conserved‐site mutation leads to largely enhanced plant lodging resistance and biomass enzymatic saccharification by reducing cellulose DP and crystallinity in rice

Genetic modification of plant cell walls has been posed to reduce lignocellulose recalcitrance for enhancing biomass saccharification. Since cellulose synthase (CESA) gene was first identified, several dozen CESA mutants have been reported, but almost all mutants exhibit the defective phenotypes in plant growth and development. In this study, the rice (Oryza sativa) Osfc16 mutant with substitutions (W481C, P482S) at P‐CR conserved site in CESA9 shows a slightly affected plant growth and higher biomass yield by 25%–41% compared with wild type (Nipponbare, a japonica variety). Chemical and ultrastructural analyses indicate that Osfc16 has a significantly reduced cellulose crystallinity (CrI) and thinner secondary cell walls compared with wild type. CESA co‐IP detection, together with implementations of a proteasome inhibitor (MG132) and two distinct cellulose inhibitors (Calcofluor, CGA), shows that CESA9 mutation could affect integrity of CESA4/7/9 complexes, which may lead to rapid CESA proteasome degradation for low‐DP cellulose biosynthesis. These may reduce cellulose CrI, which improves plant lodging resistance, a major and integrated agronomic trait on plant growth and grain production, and enhances biomass enzymatic saccharification by up to 2.3‐fold and ethanol productivity by 34%–42%. This study has for the first time reported a direct modification for the low‐DP cellulose production that has broad applications in biomass industries.     

Preparation of cross-linked carboxymethyl chitosan for repairing sciatic nerve injury in rats

A successful nerve regeneration process was achieved with nerve repair tubes made up of 1-ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) cross-linked carboxymethyl chitosan (CM-chitosan) with improved biodegradability. Chitosan has a very slow degradation rate, while the EDC cross-linked CM-chitosan tubes degraded to 30% of original weight during 8 weeks of incubation in lysozyme solution. In vitro cell culture indicated that the CM-chitosan films presented no cytotoxicity to Schwann cells. From in vivo studies using a 10 mm rat sciatic nerve defect model investigated by histomorphometry analysis, the average diameter of the fibers and the average thickness of myelin sheath in the CM-chitosan tubes were 3.7 ± 0.33 and 0.33 ± 0.04 μm, respectively, which demonstrated equivalence to nerve autografts (the current “gold” standard); furthermore, the average fiber density in the CM-chitosan tubes was 20.5 × 10³/mm², which was similar to that of autografts (21 × 10³/mm²) and significantly higher than that of common chitosan tubes (15.3 × 10³/mm²).     

A Bipartite Network-based Method for Prediction of Long Non-coding RNA–protein Interactions

As one large class of non-coding RNAs(nc RNAs), long nc RNAs(lncRNAs) have gained considerable attention in recent years. Mutations and dysfunction of lnc RNAs have been implicated in human disorders. Many lnc RNAs exert their effects through interactions with the corresponding RNA-binding proteins. Several computational approaches have been developed, but only few are able to perform the prediction of these interactions from a network-based point of view. Here,we introduce a computational method named lnc RNA–protein bipartite network inference(LPBNI). LPBNI aims to identify potential lnc RNA–interacting proteins, by making full use of the known lnc RNA–protein interactions. Leave-one-out cross validation(LOOCV) test shows that LPBNI significantly outperforms other network-based methods, including random walk(RWR)and protein-based collaborative filtering(Pro CF). Furthermore, a case study was performed to demonstrate the performance of LPBNI using real data in predicting potential lnc RNA–interacting proteins.     

The endosperm development and the variations of structures of embryo sacs: unravelling the low fecundity of Zephyranthes candida (Amaryllidaceae)

To unravel a low fecundity in Zephyranthes candida (Lindl.) Herb., the development of the endosperm was studied using conventional paraplast section technique. The results show that the endosperm develops normally and comprises four major stages viz. syncytial, cellularization, differentiation and maturation. Both proliferation of antipodal cells and their close contact with the primary endosperm nucleus were observed, which should favor transportation of nutrients and accelerate development of embryo and endosperm. In Z. candida, at least four events of nuclear migration occurred during the course of embryogenesis and endosperm development. The 12.7% structurally and functionally abnormal ovules, along with the 22.3% collapsed and aborted ovules observed accounts for the low fecundity to some extent.     

Seasonal dynamics of soil fauna in the subalpine and alpine forests of west Sichuan at different altitudes

The climate (especially temperature) often plays an important role in the structure, function as well as composition of soil organisms in different latitudes and altitudes. As one of the essential components of soil ecosystem, soil faunal community not only lays their roles as soil engineer in material cycling and energy flow, but also acts as the sensitive bio-indicator to environmental change. However, little information has been available on the responses of soil faunal community to the changed environment at different altitudes and seasons. In order to understand the seasonal dynamics of soil faunal diversity under different forests with varying altitudes, three fir (Abies faxoniana) forests were selected covering a 600 m vertical transition zone. The primary fir forest at 3600 m (A1) of altitude, mixed fir and birch forest at 3300 m (A2) of altitude, and secondary fir forest at 3000 m (A3) of altitude are representative forests in the subalpine and alpine region of west Sichuan. A 2 years study was conducted in the three subalpine and alpine forests from May in 2009 until October in 2010. Soil samples were collected in both the soil organic layer and mineral soil layer. Soil macro-fauna were picked up by hand in the fields. Meso/micro-fauna and damp living fauna were separated and collected from the soil samples by Baermann and Tullgren methods in laboratory, respectively. A total of 74,827 individuals were collected in the 2 years, belonging to seven phyla, 16 classes, 31 orders and 125 families by preliminary identification. Similar dominant groups were detected in different forests at different altitudes, consisting of Spirostreptida, Formicidae, Staphylinidae, Hesperinidae, Onychiuridae, Isotomidae, Oribatuloidae, Alicoragiidae, Secernentea, and Adenophorea. In contrast, the ordinary species of macro-fauna and the ratios of Acarina to Collembolan were obviously different. For instance, the ordinary species were dominated by Cydmaenidae and Mycetophilidae at the A1, Scaphidiidae and Helicinidae at the A2, and Lumbricida and Agelenidae at the A3, respectively. Both the individual density and the number of soil faunal groups were significantly higher in soil organic layer than those in mineral soil layer. The density and group of macro-, meso- and micro-fauna in different forests showed the order as A2 > A1 > A3, but the density of damp living fauna showed the order as A1 > A2 > A3. The functional groups of macro-fauna were mainly dominated by saprozoic. The highest density and group of macro-fauna was observed in August, while the highest value of meso/micro-fauna was detected in October. In addition, the Jacard similarity indices showed that the composition and structure of soil fauna were similar in the different forests varied with altitudes, but the Shannon–Wiener indices were significantly different. The highest values of Shannon–Wiener indices were observed in October at both the A1 and A3, and in August at the A2. The results suggested that soil faunal community kept a high diversity in the subalpine and alpine forests of west Sichuan, and their structures were significantly affected by the variation of altitudes, which provided important scientific evidences for understanding the ecological processes in the subalpine and alpine coniferous forests.     

Phosphorene as Cathode Material for High‐Voltage,Anti‐Self‐Discharge Zinc Ion Hybrid Capacitors

Output voltage and self‐discharge rate are two important performance indices for supercapacitors, which have long been overlooked, though these play a very significant role in their practical application. Here, a zinc anode is used to construct a zinc ion hybrid capacitor. Expanded operating voltage of the hybrid capacitor is obtained with novel electrolytes. In addition, significantly improved anti‐self‐discharge ability is achieved. The phosphorene‐based zinc ion capacitor exploiting a “water in salt” electrolyte with a working potential can reach 2.2 V, delivering 214.3 F g^?1 after 5000 cycles. The operating voltage is further extended to 2.5 V through the use of an organic solvent as the electrolyte; the solvent is prepared by adding 0.2 m ZnCl₂ into the tetraethylammonium tetrafluoroborate in propylene carbonate (Et₄NBF₄/PC) solvent, and it exhibits 105.9 F g^?1 even after 9500 cycles. More importantly, the phosphorene‐based capacitors possess excellent anti‐self‐discharge performance. The capacitors retain 76.16% of capacitance after resting for 300 h. The practical application of the zinc ion capacitor is demonstrated through a flexible paper‐based printed microcapacitor. It is believed that the developed zinc ion capacitor can effectively resolve the severe self‐discharge problem of supercapacitors. Moreover, high‐voltage zinc ion capacitors provide more opportunities for the application of supercapacitors.     

PPARγ与c/EBPα基因在苏太猪不同组织中的表达水平探究

为探究PPARγ与c/EBPα基因在苏太猪不同组织中的表达与脂肪沉积的关系,本实验以10月龄苏太猪为研究对象,运用实时荧光定量PCR (q RT-PCR)技术检测PPARγ与c/EBPα基因mRNA在苏太猪心、肝、脾、肺、肾、胃、背最长肌和皮下脂肪8个组织中的表达水平。结果表明,PPARγ与c/EBPα基因在苏太猪的8个组织中均有不同程度的表达,其中,PPARγ基因在苏太猪脾脏组织中的表达量最高,皮下脂肪中的表达水平仅次于脾;以背最长肌中PPARγ基因的相对表达量作对比,背最长肌与脾、肺和皮下脂肪的相对表达差异极显著(p<0.01),其余为差异不显著(p>0.05),表达量高低顺序为脾>皮下脂肪>肺>心>胃>肾>肝>背最长肌;c/EBPα基因在苏太猪的皮下脂肪的表达量最高,以背最长肌中c/EBPα基因的相对表达量作对比,在肝、脾、皮下脂肪组织中表达差异极显著(p<0.01),肺的相对表达差异显著(p<0.05),其余组织中差异不显著(p>0.05),表达量的高低顺序为皮下脂肪>肝>脾>肺>肾>心>胃>背最长肌。两基因在各组织中表达趋势趋于一致。试验结果表明PPARγ和c/EBPα基因可能对猪脂肪沉积有重要影响。     

Antimicrobial activity and secondary structure of a novel peptide derived from ovalbumin

A novel antimicrobial peptide derived from ovalbumin has been discovered. First, the peptide fragment RKIKVYLPRMK (TK9.1) was identified based on computerized predictions of the secondary structure of peptides in a protein data bank. Using HeliQuest, the sequence was developed into RKIKRYLRRMI (TK9.1.3), which was synthesized using Fmoc‐solid phase peptide synthesis, and found to have strongly antimicrobial activity against Gram‐positive and Gram‐negative bacteria, and fungi but not cytotoxic to HeLa cells and hemolysis in mouse red blood cells. Although ovalbumin itself does not have an antibacterial activity, our results suggest that it may supply the organisms that consume it with antimicrobial peptides, in support of their immunodefence.     

Distinct genomic traits of acral and mucosal melanomas revealed by targeted mutational profiling

The incidence of melanoma is rising globally including China. Comparing to Caucasians, the incidence of non‐cutaneous melanomas is significantly higher in Chinese. Herein, we performed genomic profiling of 89 Chinese surgically resected primary melanomas, including acral (n = 54), cutaneous (n = 22), and mucosal (n = 13), by hybrid capture‐based next‐generation sequencing. We show that mucosal melanomas tended to harbor more pathogenic mutations than other types of melanoma, though the biological significance of this finding remains uncertain. Chromosomal arm‐level alterations including 6q, 9p, and 10p/q loss were highly recurrent in all subtypes, but mucosal melanoma was significantly associated with increased genomic instability. Importantly, 7p gain significantly correlated with unfavorable clinical outcomes in non‐cutaneous melanomas, representing an intriguing prognostic biomarker of those subtypes. Furthermore, focal amplification of 4q12 (KIT, KDR, and PDGFRα) and RAD51 deletion were more abundant in mucosal melanoma, while NOTCH2 amplification was enriched in acral melanoma. Additionally, cutaneous melanomas had higher mutation load than acral melanomas, while mucosal melanomas did not differ from other subtypes in mutation burden. Together, our data revealed important features of acral and mucosal melanomas in Chinese including distinctive driver mutation pattern and increased genomic instability. These findings highlight the possibilities of combination therapies in the clinical management of melanoma.