Bioactivity and constituents of several common seaweeds

Bioactivity and constituents of 8 common seaweeds from Dalian intertidal zone of northern Yellow Sea were investigated. In the anti-methicillin-resistant Staphylococcus aureus (MRSA) test, Symphyocladia latiuscula and Enteromorpha intestinalis showed obvious activities with MICs much lower than 1.0 mg·mL-1 . In the DNA damage repair test (DDRT), Chondrus ocellatu showed selective inhibitory activity against the DNA repair-defective E. coli strain vs. the wild-type E. coli strain; while Sym. latiuscula, Enteromorpha intestinalis and Sar. kjellmanianum showed significant anti-E. coli activity with MICs of 64-128 g·mL-1 . In the anti-Pyricularia oryzae test, Sym. latiuscula and Rh. confervoides strongly inhibited the germination of the spores of P. oryzae on agar plate. In the brine shrimp larvicidal test, Sym. latiuscula, Rh. confervoides and Sar. kjellmanianum exhibited potent toxicity against brine shrimp larvae, with LC50 much lower than 1 mg·mL-1 . The HPTLC analysis revealed their diversified secondary metabolites. The HPLC-DAD-MS analysis of the strongest species Sym. latiuscula and database searching showed that it can produce quite diversified metabolites, including halogenated ones, some of which may be new natural products. The results demonstrated the potentials of these seaweeds in the development of new antibiotics, antitumor drugs, agricultural fungicides and pesticides.     

Visualization of protein RecR in Escherichia coli by fluorescent labelling

RecR protein, a functional equivalent of Rad52 in eukaryotes, plays a critical role in the RecF pathway of homologous recombination in Escherichia coli. By constructing and expressing the recR-yfp hybrid gene, the distribution of the RecR-YFP fusion protein was visualized in E. coli by fluorescent microscopy. Our results showed that RecR proteins can be localized predominantly in the nucleoid region of E. coli. By measuring the UV resistance of a recR mutant carrying the recR-yfp gene in the plasmid, the expressed RecR-YFP was found to be functional in improving the UV resistance of the recR deficiency strain.     

Functional genomics in the rice blast fungus to unravel the fungal pathogenicity

A rapidly growing number of successful genome sequencing projects in plant pathogenic fungi greatly increase the demands for tools and methodologies to study fungal pathogenicity at genomic scale. Magnaporthe oryzae is an economically important plant pathogenic fungus whose genome is fully sequenced. Recently we have reported the development and application of functional genomics platform technologies in M. oryzae. This model approach would have many practical ramifications in design and implementation of upcoming functional genomics studies of filamentous fungi aimed at understanding fungal pathogenicity.     

Detection of plasmid-mediated IMP-1 metallo-β-lactamase and quinolone resistance determinants in an ertapenem-resistant Enterobacter cloacae isolate

Objective: To investigate the mechanism of carbapenem resistance and the occurrence of plasmid-mediated quinolone resistance determinants qnr and aac(6′)-Ib-cr in a clinical isolate of Enterobacter cloacae. Methods: An ertapenem-resistant E. cloacae ZY106, which was isolated from liquor puris of a female gastric cancer patient in a Chinese hospital, was investigated. Antibiotic susceptibilities were determined by agar dilution method. Conjugation experiments, isoelectric focusing, polymerase chain reaction (PCR), and DNA sequence analyses of plasmid-mediated carbapenemases and quinolone resistance determinants were preformed to confirm the genotype. Outer membrane proteins (OMPs) were examined by urea-sodium dodecyl sulfatepolyacrylamide gel electrophoresis (Urea-SDS-PAGE). Results: Minimum inhibitory concentrations (MICs) of imipenem, meropenem, and ertapenem for ZY106 were 2, 4, and 16 μg/ml, respectively. Conjugation studies with Escherichia coli resulted in the transfer of significantly reduced carbapenem susceptibility. ZY106 produced IMP-1 metallo-β-lactamase and CTX-M-3 extended-spectrum β-lactamase, and E. coli transconjugant produced IMP-1. Plasmid-mediated quinolone resistance determinant qnrS1 was detected in ZY106. Transfer of the qnrS1-encoding-plasmid into E. coli by conjugation resulted in intermediate resistance to ciprofloxacin in E. coli transconjugant. Urea-SDS-PAGE analysis of OMPs showed that ZY106 lacked an OMP of approximately 38 kDa. Conclusion: It is the first IMP-1-producing Enterobacteriaceae in China and the first report of a clinical isolate that harbors both blaIMP and qnrS genes as well. The blaIMP-1, blaCTX-M-3, and qnrS1 are encoded at three different plasmids. IMP-1 combined with the loss of an OMP possibly resulted in ertapenem resistance and reduced imipenem and meropenem susceptibility in E. cloacae.     

Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum

Agrobacterium tumefaciens-mediated transformation （ATMT） system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 106 conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction （PCR） amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.     

Development of a novel conductance-based technology for environmental bacterial sensing

In this study, a simple impedance based technology for measuring bacterial concentrations was developed. The measurement system includes the signal amplification, copper probes and a sample loader. During the experiments, the conductance of Bacillus subtilis var niger, Pseudomonas fluorescens, and Escherichia coli were measured using the combination of a pre-amplifier and a lock-in amplifier. The conductance data were modeled verses the bacterial concentrations. Results indicated that the relationship between the conductance of bacterial suspensions and their concentrations follows a generic model: $Y = C_1 + C_2 \times e^{\left( { - X/C_3 } \right)}$ , where Y is the conductance (S), X is the bacterial concentration (Number/mL: abbreviated to N/mL) for all species tested, and C _1–3 are constants. Gram negative P. fluorescens and E. coli assumed similar conductance curves, which were flatter than that of gram positive B. subtilis var niger. For P. fluorescens and E. coli the culturing technique resulted in higher concentration levels (statistically significant) from 2 to 4 times that measured by the impedance based technology. For B. subtilis var niger, both methods resulted in similar concentration levels. These differences might be due to membrane types, initial culturability and the obtained conductance curves. The impedance based technology here was shown to obtain the bacterial concentration instantly, holding broad promise in realtime monitoring biological agents.     

Effective antibacterial and phase change PEG/Cu2O@A-HNTs composites for melt-spun difunctional PA6 fiber

Serving as the only defensive line between pathogens and human body, personal protective equipment (PPE) is increasingly attractive among researchers because of their strong antibacterial and temperature control abilities. However, efficient antibacterial properties and regenerative thermal control simultaneously remain unexplored in PPE. Here, one-dimensional halloysite nanotubes (HNTs) modified by acid etch method, are used to synthesis a multifunctional material of PEG/Cu₂O@A-HNTs via in-situ reduction and physical adsorption which serves the above two purposes. PEG/Cu₂O@A-HNTs showed rapid bacterial inactivation and achieved 96.3% bacteriostatic rate against E. coli in only 20 ?min. Meanwhile a broad and complete inactivation spectrum included both E. coli and S. aureus following a series of antibacterial mechanisms. Moreover, adsorption of 70 ?wt% PEG by acid etch HNTs is attributed to the nearly 3 times increased specific surface area compared with native HNTs. This enabled PEG/Cu₂O@A-HNTs to attain a phase change enthalpy of 108.4 ?J/g. In addition, using PEG/Cu₂O@A-HNTs as additives, antibacterial and phase change fiber (APCF) were melt-spun. Their efficiency factor against E. coli and S. aureus was above 99.99%, and retained a temperature control ability for 180s and 272s compared with PA6 fiber in hot and frigid environments respectively.     

一株尿路致病性大肠杆菌噬菌体的分离鉴定及生物学特性分析

本研究从尿路感染患者尿液样本中分离、鉴定大肠杆菌裂解性噬菌体,对其裂菌活性等生物学特性进行分析,为探索应用噬菌体治疗耐药性尿路致病性大肠杆菌导致的尿路感染奠定基础.利用双层琼脂平板法,从尿液样品中分离、纯化,得到一株能裂解大肠杆菌的噬菌体.通过电镜观察噬菌体形态,测定其裂菌谱、最佳感染复数、一步生长曲线、以及热稳定性、...     

Sprayed Pickering emulsion with high antibacterial activity for wound healing

Effective treatment of wound injuries can prevent infections and promote healing. In this work, a sprayed Pickering emulsion composed of chitosan nanoparticles, tea tree oil and curcumin was formulated. The shear thinning behaviour allowed this Pickering emulsion to spray even with a high viscosity provided by the aqueous phase containing chitosan nanoparticles. Moreover, the Pickering emulsion could recover to the high viscosity after spraying onto the wounds, leading to the long residence time and intimate contact. In addition, the Pickering emulsion provided a carrier to encapsulate the tea tree oil, which caused sustained-release effects and further strengthened tea tree oil's antibacterial activity towards the Gram-negative Escherichia coli (E. coli), Gram-positive Staphylococcus aureus (S. aureus) and Gram-positive methicillin-resistant Staphylococcus aureus (MRSA). Besides, this multi-phase system offered a platform to load with hydrophobic curcumin, leading to better synergistic healing effects. Meanwhile, the Pickering emulsion avoided the irritation and toxicity related to surfactants, which was far safer than the classical emulsion especially when used in the defective skin. All these excellent properties endowed this sprayed Pickering emulsion with promising prospects for wound healing.     

High-quality reference genes for quantifying the transcriptional responses of Oryza sativa L. (ssp. indica and japonica) to abiotic stress conditions

Rice (Oryza sativa L.) is important to food security and is also an excellent model plant for numerous cereal crops. A functional genomics study in rice includes characterization of the expression dynamics of genes by quantitative real-time PCR (qPCR) analysis; this is a significant key for developing rice varieties that perform well in the face of adverse climate change. The qPCR analysis requires the use of appropriate reference genes in order to make any quantitative interpretations meaningful. Here, the new potential reference genes were selected from a huge public database of rice microarray experiments. The expression stability of 14 candidates and 4 conventional reference genes was validated by geNorm PLUS and NormFinder software. Seven candidates are superior to the conventionally used reference genes in qPCR and three genes can be used reliably for quantitating the expression of genes involved in abiotic stress responses. These high-quality references EP (LOC_Os05g08980), HNR (LOC_Os01g71770), and TBC (LOC_Os09g34040) worked very well in three indica genotypes and one japonica genotype. One of indica genotypes including the Jasmine rice, KDML105 developed in Thailand for which no reference genes have been reported until now.     

The stress response factor RpoS is required for the natural transformation of Escherichia coli

The natural transformation of Escherichia coli is a novel and recently developed system that has significance for genetic studies and the biological safety of genetic engineering. However, the mechanisms of transformation, including development of competence and DNA uptake, are not thoroughly understood. In this study, we demonstrated the effect of the general stress response regulator RpoS, which has been associated with E. coli transformation, on natural transformation performed in an “open system”. We find that RpoS is required for natural transformation but not to artificial transformation and RpoS mainly affect transformation in the liquid culture prior to plating. In the liquid culture, RpoS over-expression promotes natural transformation in early exponential phase and static incubation accumulates RpoS and promotes transformation to a limited extent. These findings provide detailed understanding of RpoS function on natural transformation.     

Expression and purification of bioactive high-purity human midkine in Escherichia coli

Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-β-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells.     

Insights on evolution of virulence and resistance from the whole-genome analysis of a predominant methicillin-resistantStaphylococcus aureus clone sequence type 239 in China

Earlier, we reported that ST239 was the 15-year predominant methicillin-resistant Staphylococcus aureus （MRSA） clone in China. In this study, MRSA strain CN79 belonging to ST239 and isolated from blood was used to determine the whole tive genomics analysis was genome sequence. Compara- done between MRSA CN79 and 25 sequenced S. aureus in the NCBI GenBank data- base. A total of 2,734 protein-encoding genes were iden- tified in the MRSA CN79 genome, which carries 11 antibiotic resistance genes and 65 virulence genes. Two prophages phiCN79A and phiNM3-1ike were found on the MRSA CN79 genome. MRSA CN79 carries 30 specific genes that are absent from the 25 sequenced S. aureus genomes. Most of them were prophage-related genes. Several antibiotic resistance genes, such as [3-1actamase and ABC-type multidrug transport system gene, were located on the genomic island vSal3. The antibiotic resis- tance genes, such as tet （M）, ermA1, and blaZ, were also located on different transposons. The virulence genes sea,map, hlb, and sak are located on phiNM3-1ike prophage and the exotoxin genes are carried on the genomic island vSaa. These results suggest that ST239 MRSA strains are widespread owing to horizontal acquisition of the mobile genetic elements harbored antibiotic resistance genes and virulence genes in response to environmental selective pressures, such as antibiotics and the human immune sys- tem during evolution.     

Development of a propulsion system for a biomimetic thruster

We studied theoretically and experimentally a biomimetic propulsion system inspired by the motility mechanisms of bacteria such as E. coli. Our goal was to investigate the effect of the ??complex?? filament of Rhizobium Meliloti bacteria on thrust force. The complex filament is a helically perturbed filament, similar to a plain filament threaded through a small helix. The propulsive performance of this system was estimated by modeling the dynamics of helical wave motion in viscous fluid. The model consists of a helical filament which is axially rotated at angular velocity ??. Resistive force theory (RFT) was applied to this model to calculate the thrust force and required torque. The Buckingham PI theorem (non-dimensional analysis) was also used to analyze the theoretical results. The procedure for making a complex filament with various pitch angles ?? _s from a small helix and plain filament is explained in detail. To validate the theoretical results for helical wave propulsion and compare the characteristics of complex and plain filaments together, an experiment was performed to measure the thrust forces in silicone oil. The experimental results agreed with the theoretical values predicted by RFT. The thrust forces of complex filaments depended on the shape of small helix winding. The maximum thrust force was achieved at a small helix pitch angle of ?? _s = 45°. In addition, we found that the thrust force generated by a complex filament had a value about 10% higher than that of a plain filament with the same equivalent diameter d _e .     

中国三个不同地区的活羊调运网络及其布鲁氏菌病传播风险

【目的】揭示不同地区的活羊调运网络结构,对比其布鲁氏菌病疫情暴发风险大小,并找出其防控关键点。【方法】构建内蒙古3个不同地区的活羊调运社会网络并进行社会网络分析,采用Uninet6.0绘制网络关系图及计算相关的网络测度。【结果】农区活羊调运网络为网状结构,而牧区活羊调运网络呈现许多"雪花状"结构,半农半牧区的活羊调运网络则兼具农区和牧区的网络特征;农区和半农半牧区的活羊调运网络表现出"小世界"的特征;农区传播力较强的节点大多为养殖户,而贩运经纪人在牧区和半农半牧区网络中具有重要的传播影响力。【结论】农区和半农半牧区的布鲁氏菌病暴发风险较高;社会网络分析与动物调运追踪系统结合能更好地作为布鲁氏菌病防控的支撑,在疫情暴发时冻结关键节点的活羊流通活动能够有效控制疫情。     

The induction of Sinorhizobium meliloti C4-dicarboxylate transport system （Dct） is regulated by oxygen concentration

The Sinorhizobium meliloti C4-dicarboxylate transport （Dct） system is essential for symbiotic nitrogen fixation. The dctA gene, encoding the C4-dicarboxylate permease, is expressed in both free living and symbiotic cells. But in free living cells expression of dctD and dctB is absolutely required for the expression of dctA. In this study, in order to investigate the effect of oxygen concentration on the induction of Dct system, E. coli DH5a strain which carries the plasmid-encoded dctABD operon was used in tube assays. It was found that the specific induction of Dct system oc- curred only at a certain depth under the surface of M63-0.6% agar media, suggesting that Dct system could respond to oxygen concentration during succinate-induced expression. Furthermore, when measured at different oxygen concentrations, the highest expression level was observed at oxygen concentration of 2%. Thus, we predict that in addition to dicarboxylates, the induction of Dct system may also regulated by oxygen concentration.     

Synthesis, characterization and antibacterial activity of copper, nickel and bimetallic Cu– Ni nanoparticles for potential use in dental materials

The antibacterial effect is a desirable property in dental materials.Development of simple methods for the preparation of nanosized metal particles has attracted significant attention because of their future applications due to unusual size-dependent antibacterial properties.Copper(Cu),Nickel(Ni) and bimetallic Cu–Ni nanoparticles were prepared by a simple chemical method and their antibacterial activity was tested against the widely used standard human pathogens Staphylococcus aureus(gram-negative) and Escherichia coli(gram-positive).Additionally,these nanoparticles were tested against the dental pathogen Streptococcus mutans.Our results are promising for potential use in dental materials science.     

The in vitro interaction of CmeA with CmeC

The membrane fusion protein CmeA and the outer membrane channel CmeC,two important components of CmeABC system in Campylobacter jejuni,were expressed in Escherichia coli.After Ni-NTA and ion exchange columns purification,size exclusion chromatography showed that CmeA primarily existed as trimer and CmeC existed as monomer.Then the interaction between CmeA and CmeC was analyzed by dithiobis(succinimidyl propionate)(DSP)chemical crosslinking,pull-down assay on a Ni-NTA column,and isothermal titration calorimetry(ITC)measurement.The results clearly showed the CmeA–CmeC complex band,which confirmed the interaction in vitro and this interaction is independent of substrate and CmeB.It suggests that the mechanism underlying CmeABC function in C.jejuni is similar to that of AcrABTolC in E.coli.