大肠杆菌topA－菌株中质粒pBR322的超凝集结构

超凝集DNA是质粒DNA的一种特殊拓扑结构形式，最初在大肠杆菌SD108 (topA⁺ gyrB225)细胞中被发现. 现在大肠杆菌DM800(topA^－ gyrB225)细胞中也发现了这种结构，这说明超凝集DNA的形成与细胞内旋转酶活性降低有直接关系，而与拓扑异构酶Ⅰ的存在与否无关. 体外实验的结果显示，具有很强的正超螺旋松弛活性的拓扑异构酶Ⅳ可以将超凝集DNA完全松弛，这也证明质粒DNA的超凝集结构与超螺旋结构在细胞内是可以互变的. 使用原子力显微镜对分离到的pBR322 DNA超凝集结构进行分析，并与普通超螺旋进行比较，结果表明，超凝集DNA分子的结构发生了巨大的变化，其分子长度比正常超螺旋分子缩短了约30%，宽度和高度则增加了60%，结构更接近于A型DNA. 另外，原子力显微镜研究结果表明，氯喹的嵌入并非改变了超凝集DNA的超螺旋状态，而是使其打结并最终压缩成一团.     

Vinealobryum guangdongensis (Pottiaceae,Musci), a new species with multicellular axillary gemmae from Guangdong,China

A new species belonging to Didymodon sensu lato, Vinealobryum guangdongensis, is described and illustrated from Nanling National Forest Park of Guangdong, China. It is characterized by noteworthily thick‐walled cells of the cauline central cylinder, ovate‐lanceolate leaves that are appressed when dry, acuminate to acute leaf apices, leaf base abruptly broadened and quickly narrowed to the insertion, leaf margins recurved in proximal 2/3 to 3/4, short‐excurrent costa with 0–1 layer of ventral stereids, laminal cells with conical or elliptical papillae either over the lumina or over transverse walls, presence of gemmae in the leaf axils, and KOH laminal color reaction red to reddish orange. This new species is compared with the most similar species and its ecology is discussed.     

Soil micro-food web interactions and rhizosphere priming effect

Background

Seeds host bacterial inhabitants but only a limited knowledge is available on which taxa inhabit seed, which niches could be colonized, and what the routes of colonization are.

Scope

Within this commentary, a discussion is provided on seed bacterial inhabitants, their taxa, and from where derive the seed colonizers.

Conclusions

Seeds/and grains host specific bacteria deriving from the anthosphere, carposphere, or from cones of gymnosperms and inner tissues of plants after a long colonization from the soil to reproductive organs.

     

<Emphasis Type="Italic">PpVIN2</Emphasis>, an acid invertase gene family member,is sensitive to chilling temperature and affects sucrose metabolism in postharvest peach fruit

We previously reported that expression and activity of acid invertases (AI) are increased in peach fruit under chilling stress. In order to determine which AI genes respond to chilling stress, seven AI genes, two vacuolar invertases (VINs) and five cell wall-bound invertases (CWINs), were identified and cloned. The predicted amino acid sequences of the genes contain conserved sites characteristic of plant AIs such as NDPNG/A, the sucrose-binding site, and MWECV/P, a cysteine catalytic motif. Using gene-specific primers, the expression of each gene was measured in ‘Baifeng’ and ‘Yulu’ peach fruits stored at 0, 5, 10 and 20 °C. Of the seven genes, expression of PpVIN2 was the most affected by chilling stress; the largest increases were in fruit stored at 5 °C, up to 17-fold in ‘Baifeng’ fruit, and up to 280-fold in ‘Yulu’ fruit. Overall, VIN activity was much higher than CWIN activity in stored peach fruit. In both cultivars reducing sugar content increased significantly and sucrose content decreased gradually during storage at 5 °C relative to other temperatures, and was accompanied by severe chilling injury symptoms. Thus, PpVIN2 appears to be induced by chilling and may play an important role in sucrose metabolism in peach fruit subjected to cold storage.     

Eutectic‐Derived Mesoporous Ni‐Fe‐O Nanowire Network Catalyzing Oxygen Evolution and Overall Water Splitting

The development of efficient and abundant water oxidation catalysts is essential for the large‐scale storage of renewable energy in the form of hydrogen fuel via electrolytic water splitting, but still remains challenging. Based upon eutectic reaction and dealloying inheritance effect, herein, novel Ni‐Fe‐O‐based composite with a unique mesoporous nanowire network structure is designed and synthesized. The composite exhibits exceptionally low overpotential (10 mA cm^?2 at an overpotential of 244 mV), low Tafel slope (39 mV dec^?1), and superior long‐term stability (remains 10 mA cm^?2 for over 60 h without degradation) toward oxygen evolution reaction (OER) in 1 m KOH. Moreover, an alkaline water electrolyzer is constructed with the Ni‐Fe‐O composite as catalyst for both anode and cathode. This electrolyzer displays superior electrolysis performance (affording 10 mA cm^?2 at 1.64 V) and long‐term durability. The remarkable features of the catalyst lie in its unique mesoporous nanowire network architecture and the synergistic effect of the metal core and the active metal oxide, giving rise to the strikingly enhanced active surface area, accelerated electron/ion transport, and further promoted reaction kinetics of OER.     

Identification of a Receptor-Binding Domain in the S Protein of the Novel Human Coronavirus Middle East Respiratory Syndrome Coronavirus as an Essential Target for Vaccine Development

A novel human Middle East respiratory syndrome coronavirus (MERS-CoV) caused outbreaks of severe acute respiratory syndrome (SARS)-like illness with a high mortality rate, raising concerns of its pandemic potential. Dipeptidyl peptidase-4 (DPP4) was recently identified as its receptor. Here we showed that residues 377 to 662 in the S protein of MERS-CoV specifically bound to DPP4-expressing cells and soluble DPP4 protein and induced significant neutralizing antibody responses, suggesting that this region contains the receptor-binding domain (RBD), which has a potential to be developed as a MERS-CoV vaccine.     

Effective antimicrobial activity of Cbf‐K16 and Cbf‐A7A13 against NDM‐1‐carrying Escherichia coli by DNA binding after penetrating the cytoplasmic membrane in vitro

New Delhi metallo‐beta‐lactamase‐1(NDM‐1)‐carrying isolates, which are resistant to most clinical used antibiotics except for tigecycline and colistin, have been found worldwide. Cathelicidin‐BF (BF‐30) is found in the venom of the snake Bungarus fasciatus and exhibits broad antimicrobial activity. Cbf‐K₁₆ and Cbf‐A₇A₁₃ were obtained by mutating Lys¹⁶, Ala⁷, and Ala¹³ of BF‐30, respectively. To investigate their antimicrobial activities against NDM‐1 carrying bacteria, recombinant Escherichia coli BL21 (DE3)‐NDM‐1 with high NDM‐1 activity was constructed by inserting the Klebsiella pneumoniae NDM‐1 gene (GenBank accession no. HQ328085) into a pET28a vector and transforming it into E. coli BL21 (DE3). The peptides showed effective antimicrobial activities against NDM‐1‐carrying E. coli, and the minimum inhibitory concentrations of Cbf‐K₁₆ and Cbf‐A₇A₁₃ were only 4 and 8 µg/ml, whereas those of minimum bactericidal concentrations were 8 and 16 µg/ml, respectively. A time course experiment showed that colony forming unit counts rapidly decreased, and bacteria were thoroughly eliminated within 3 and 6 h by the Cbf‐K₁₆ and Cbf‐A₇A₁₃ treatments, respectively. The peptides penetrated the bacterial cell membrane and enabled β‐galactosidase leakage, and caused the cytoplasmic membrane to become permeable, and finally bound to the DNA. The genomic DNA of E. coli was completely unable to migrate on an agarose gel after Cbf‐K₁₆ treatment (8 µg/ml). These data demonstrated that Cbf‐K₁₆ and Cbf‐A₇A₁₃ possess effective antimicrobial activity against drug‐resistant strains, including NDM‐1 carrying E. coli BL21 (DE3)‐NDM‐1, by binding to DNA after penetrating the cytoplasmic membrane in vitro, which may have potential therapeutic value for the treatment of NDM‐1‐carrying bacterial infections. Copyright © 2013 European Peptide Society and John Wiley & Sons, Ltd.     

Annexin A1 mediates the anti‐adhesive effects of dexamethasone during the cell–cell interaction between the all‐trans retinoic acid‐treated acute promyelocytic leukemic cells and endothelial cells

Annexin A1 (AnxA1) is an important anti‐inflammatory mediator during granulocytic differentiation in all trans‐retinoic acid (ATRA) treated acute promyelocytic leukemic (APL) cells. Dexamethasone has been used successfully to prevent complications in ATRA‐treated APL patients, although its mechanism of action is still not clear. In the present study, we have examined the effect of dexamethasone on the modulation of AnxA1 in ATRA‐APL NB4 (ATRA‐NB4) cells, ATRA‐NB4 cells‐derived microparticles (MPs) and its role during cell–cell interaction between ATRA‐NB4 cells and endothelial cells. Our results have shown that dexamethasone can inhibit the percentage of ATRA‐NB4 cells expressing surface AnxA1 and its receptor FPR2/ALX in a time‐dependent manner based on flow cytometric analysis. However, dexamethasone treatment of ATRA‐NB4 cells has no significant effect on the level of AnxA1 mRNA, the total cellular level of AnxA1 protein or the release of AnxA1 from these cells, as determined by RT‐PCR, Western blotting, and ELISA, respectively. Further studies demonstrate that dexamethasone is able to significantly inhibit the adhesion of ATRA‐NB4 cells to endothelial cells, and this anti‐adhesive effect can be inhibited if the cells were pre‐treated with a neutralizing antibody specific for AnxA1. Finally, dexamethasone also enhances the release of AnxA1‐containing MPs from ATRA‐NB4 cells which can in turn prevent the adhesion of the ATRA‐NB4 cells to endothelial cells. We conclude that biologically active AnxA1 originating from dexamethasone‐treated ATRA‐APL cells and their MPs plays an anti‐adhesive effect and this contributes to inhibit the adhesion of ATRA‐APL cell to endothelial cells. J. Cell. Biochem. 114: 551–557, 2013. © 2012 Wiley Periodicals, Inc.     

Study on the interaction between a water-soluble dinuclear nickel complex and bovine serum albumin by spectroscopic techniques

The interaction of a water-soluble dinuclear nickel(II) complex, [Ni₂(EGTB)(CH₃CN)₄](ClO₄)₄·4H₂O (EGTB = ethylene glycol-bis(β-aminoethyl ether) N,N,N′,N′-tetrakis(2-benzimidazoyl)) (1), and bovine serum albumin (BSA) was investigated under physiological conditions using fluorescence, synchronous fluorescence, UV–vis absorption and circular dichroism (CD). The experimental results suggested that the nickel(II) complex could bind to BSA with binding constant (K) ~ 10⁴ M^?1 and quench the intrinsic fluorescence of BSA through a static quenching mechanism. The thermodynamic parameters, ΔG°, ΔH°, and ΔS°, calculated at different temperatures, indicated that the binding reaction was spontaneous and electrostatic interactions played a major role in this association. Based on the number of binding sites, it was considered that one molecule of complex 1 could bind to a single site or two sites of the BSA molecule or the two binding modes coexisted. In view of the results of site marker competition experiments, the reactive sites of BSA to complex 1 mainly located in subdomain IIA (site I) and subdomain IIIA (site II) of BSA. Moreover, the binding distance, r, between donor (BSA) and acceptor (complex 1) was 5.13 nm according to Förster nonradiation energy transfer theory. Finally, as shown by the UV–vis absorption, synchronous fluorescence and CD, complex 1 could induce conformation and microenvironmental changes of BSA. The results obtained herein will be of biological significance in toxicology investigation and anticancer metallodrug design.