Identification of Cj1051c as a Major determinant for the restriction barrier of Campylobacter jejuni StrainNCTC11168

Campylobacter jejuni is a leading cause of human diarrheal illness in the world, and research on it has benefitted greatly by the completion of several genome sequences and the development of molecular biology tools. However, many hurdles remain for a full understanding of this unique bacterial pathogen. One of the most commonly used strains for genetic work with C. jejuni is NCTC11168. While this strain is readily transformable with DNA for genomic recombination, transformation with plasmids is problematic. In this study, we have identified a determinant of this to be cj1051c, predicted to encode a restriction-modification type IIG enzyme. Knockout mutagenesis of this gene resulted in a strain with a 1,000-fold-enhanced transformation efficiency with a plasmid purified from a C. jejuni host. Additionally, this mutation conferred the ability to be transformed by plasmids isolated from an Escherichia coli host. Sequence analysis suggested a high level of variability of the specificity domain between strains and that this gene may be subject to phase variation. We provide evidence that cj1051c is active in NCTC11168 and behaves as expected for a type IIG enzyme. The identification of this determinant provides a greater understanding of the molecular biology of C. jejuni as well as a tool for plasmid work with strain NCTC11168. © 2012, American Society for Microbiology.

Factors affecting seismic response of submarine slopes

The response of submerged slopes on the continental shelf to seismic or storm loading has become an important element in the risk assessment for offshore structures and "local" tsunami hazards worldwide. The geological profile of these slopes typically includes normally consolidated to lightly overconsolidated soft cohesive soils with layer thickness ranging from a few meters to hundreds of meters. The factor of safety obtained from pseudo-static analyses is not always a useful measure for evaluating the slope response, since values less than one do not necessarily imply slope failure with large movements of the soil mass. This paper addresses the relative importance of different factors affecting the response of submerged slopes during seismic loading. The analyses use a dynamic finite element code which includes a constitutive law describing the anisotropic stress-strain-strength behavior of normally consolidated to lightly overconsolidated clays. The model also incorporates anisotropic hardening to describe the effect of different shear strain and stress histories as well as bounding surface principles to provide realistic descriptions of the accumulation of the plastic strains and excess pore pressure during successive loading cycles. The paper presents results from parametric site response analyses on slope geometry and layering, soil material parameters, and input ground motion characteristics. The predicted maximum shear strains, permanent deformations, displacement time histories and maximum excess pore pressure development provide insight of slope performance during a seismic event. © 2006 Author(s). This work is licensed under a Creative Commons License.

Seismic response of submarine slopes

The geological profile of submerged slopes on the continental shelf typically includes soft cohesive soils with thicknesses ranging from a few meters to tens or hundreds of meters. The response of these soils in simple shear tests is largely influenced by the presence of an initial consolidation shear stress, inducing anisotropic stress-strain-strength properties which depend also on the direction of shear. In this paper, a new simplified effective-stress-based model describing the behavior of normally to lightly overconsolidated cohesive soils is used in conjunction with a one-dimensional seismic site response analysis computer code to illustrate the importance of accounting for anisotropy and small strain nonlinearity. In particular, a simple example is carried out to compare results for different slope inclinations. Depth profiling of the maximum shear strains and permanent deformations provide insight into the mechanisms of deformation during a seismic event, and the effects of sloping ground conditions.

Modeling cyclic behavior of lightly overconsolidated clays in simple shear

Assessment of seismic performance and estimation of permanent displacements for submerged slopes require the accurate description of the soil's stress-strain-strength relationship under irregular cyclic loading. The geological profile of submerged slopes on the continental shelf typically consists of normally to lightly overconsolidated clays with depths ranging from a few meters to a few hundred meters and very low slope angles. This paper describes the formulation of a simplified effective-stress-based model, which is able to capture the key aspects of the cyclic behavior of normally consolidated clays. The proposed constitutive law incorporates anisotropic hardening and bounding surface principles to allow the user to simulate different shear strain and stress reversal histories as well as provide realistic descriptions of the accumulation of plastic shear strains and excess pore pressure during successive loading cycles. (C) 2000 Published by Elsevier Science Ltd. | Assessment of seismic performance and estimation of permanent displacements for submerged slopes require the accurate description of the soil's stress-strain-strength relationship under irregular cyclic loading. The geological profile of submerged slopes on the continental shelf typically consists of normally to lightly overconsolidated clays with depths ranging from a few meters to a few hundred meters and very low slope angles. This paper describes the formulation of a simplified effective-stress-based model, which is able to capture the key aspects of the cyclic behavior of normally consolidated clays. The proposed constitutive law incorporates anisotropic hardening and bounding surface principles to allow the user to simulate different shear strain and stress reversal histories as well as provide realistic descriptions of the accumulation of plastic shear strains and excess pore pressures during successive loading cycles.

Lymph Node Colonization Dynamics after Oral Salmonella Typhimurium Infection in Mice

An understanding of how pathogens colonize their hosts is crucial for the rational design of vaccines or therapy. While the molecular factors facilitating the invasion and systemic infection by pathogens are a central focus of research in microbiology, the population biological aspects of colonization are still poorly understood. Here, we investigated the early colonization dynamics of Salmonella enterica subspecies 1 serovar Typhimurium (S. Tm) in the streptomycin mouse model for diarrhea. We focused on the first step on the way to systemic infection - the colonization of the cecal lymph node (cLN) from the gut - and studied roles of inflammation, dendritic cells and innate immune effectors in the colonization process. To this end, we inoculated mice with mixtures of seven wild type isogenic tagged strains (WITS) of S. Tm. The experimental data were analyzed with a newly developed mathematical model describing the stochastic immigration, replication and clearance of bacteria in the cLN. We estimated that in the beginning of infection only 300 bacterial cells arrive in the cLN per day. We further found that inflammation decreases the net replication rate in the cLN by 23%. In ccr7-/- mice, in which dendritic cell movement is impaired, the bacterial migration rate was reduced 10-fold. In contrast, cybb-/- mice that cannot generate toxic reactive oxygen species displayed a 4-fold higher migration rate from gut to cLN than wild type mice. Thus, combining infections with mixed inocula of barcoded strains and mathematical analysis represents a powerful method for disentangling immigration into the cLN from replication in this compartment. The estimated parameters provide an important baseline to assess and predict the efficacy of interventions. © 2013 Kaiser et al.

A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae

Isolation of high neutral lipid-containing microalgae is key to the commercial success of microalgae-based biofuel production. The Nile red fluorescence method has been successfully applied to the determination of lipids in certain microalgae, but has been unsuccessful in many others, particularly those with thick, rigid cell walls that prevent the penetration of the fluorescence dye. The conventional "one sample at a time" method was also time-consuming. In this study, the solvent dimethyl sulfoxide (DMSO) was introduced to microalgal samples as the stain carrier at an elevated temperature. The cellular neutral lipids were determined and quantified using a 96-well plate on a fluorescence spectrophotometer with an excitation wavelength of 530 nm and an emission wavelength of 575 run. An optimized procedure yielded a high correlation coefficient (R-2 = 0.998) with the lipid standard triolein and repeated measurements of replicates. Application of the improved method to several green algal strains gave very reproducible results with relative standard errors of 8.5%, 3.9% and 8.6%, 4.5% for repeatability and reproducibility at two concentration levels (2.0 mu g/mL and 20 mu g/mL), respectively. Moreover, the detection and quantification limits of the improved Nile red staining method were 0.8 mu g/mL and 2.0 mu g/mL for the neutral lipid standard triolein, respectively. The modified method and a conventional gravimetric determination method provided similar results on replicate samples. The 96-well plate-based Nile red method can be used as a high throughput technique for rapid screening of a broader spectrum of naturally-occurring and genetically-modified algal strains and mutants for high neutral lipid/oil production. (C) 2009 Published by Elsevier B.V.

Comparative studies on physiological responses to phosphorus in two phenotypes of bloom-forming Microcystis

Toxic Microcystis blooms frequently occur in eutrophic water bodies and exist in the form of colonial and unicellular cells. In order to understand the mechanism of Microcystis dominance in freshwater bodies, the physiological and biochemical responses of unicellular ( 4 strains) and colonial ( 4 strains) Microcystis strains to phosphorus ( P) were comparatively studied. The two phenotype strains exhibit physiological differences mainly in terms of their response to low P concentrations. The growth of four unicellular and one small colonial Microcystis strain was significantly inhibited at a P concentration of 0.2 mg l - 1; however, that of the large colonial Microcystis strains was not inhibited. The results of phosphate uptake experiments conducted using P- starved cells indicated that the colonial strains had a higher affinity for low levels of P. The unicellular strains consumed more P than the colonial strains. Alkaline phosphatase activity in the unicellular strains was significantly induced by low P concentrations. Under P- limited conditions, the oxygen evolution rate, Fv/ Fm, and ETRmax were lower in unicellular strains than in colonial strains. These findings may shed light on the mechanism by which colonial Microcystis strains have an advantage with regard to dominance and persistence in fluctuating P conditions.

Strain Effects on the Optical Polarization Properties of R-Plane Wurtzite GaN

Using the effective-mass Hamiltonian for an arbitrary direction wurtzite semiconductor on the basis of k.p theory, we investigate the strain effects on the transition energies and optical properties in the R-plane ([1012]-oriented plane) GaN. The results show that (1) the transition energies decrease with the biaxial strains changing from -0.5 to 0.5%; and (2) giant optical anisotropy appears in the R-plane which is significantly affected by the biaxial strains. We clarify the relation between the strains and the polarization properties. Finally, we discuss the application of these properties to the R-plane GaN based devices. (c) 2009 The Japan Society of Applied Physics

INPLANE X-RAY-SCATTERING OF EPITAXIAL STRUCTURES

A new approach for in-plane X-ray scattering from the cleavages of epitaxial films or superlattices, where the scattering vectors are parallel to the interfaces, is proposed. This method can be employed to determine directly the in-plane X-ray strains and other atomic registry along the interfaces of the epitaxial structures.

染料脱色菌株筛选及染料脱色作用研究

染料废水成分复杂，色度高，是主要的环境污染源之一。应用微生物对染料废水脱色具有处理费用低和操作管理方便等优势，具有较好的发展前景。 本文采用筛选到一株短刺小克银汉霉(Cuuninghamella echinulata) KM3及一株红酵母(Rhodotorula sp.)YR-1分别对偶氮染料刚果红废水和酸性红废水进行了脱色研究，同时利用KM3及YR-1组成的复合菌种脱色体系对酸性红废水进行了脱色实验，为染料脱色真菌的应用奠定了基础。 KM3形成的菌丝球对多种染料具有较好脱色效果。KM3最适菌丝球形成条件：pH6-7、装液量90ml、孢子悬液浓度105个/ml (接种5ml)、摇床转速120rpm、培养温度30℃、培养时间72h；成球机理为凝聚型发育。KM3菌丝球对刚果红最佳脱色条件为：染料溶液初始pH值7.0，温度为33℃，摇床转速为120rpm。在刚果红50~200mg/l 浓度范围内， KM3菌丝球对刚果红吸附脱色动力学可用拟二级动力学方程描述。菌丝球对刚果红的吸附行为符合Langmuir和Freundlich 等温方程模型，Langmuir方程能更好的描述菌丝球对刚果红的吸附行为。丙酮对刚果红的解吸率最高，其解吸动力学过程符合二级动力学方程。将KM3应用在真菌生物滤床反应器中，在非灭菌和灭菌情况下接种孢子悬液后处理刚果红废水的过程中，均需长时间挂膜，处理效果一般。反应器中投加菌丝球后处理刚果红废水在3小时内达到92%以上的脱色效果。 YR-1对酸性红脱色机制属于降解脱色。YR-1对酸性红的最佳脱色培养条件为：初始pH 5.0、接种量5%、温度30℃；最佳碳源是葡萄糖，其次是蔗糖；最佳氮源是酵母膏或蛋白胨。在替换分批脱色实验和连续补料脱色实验中，脱色率一直保持在75%以上，显示YR-1在上述脱色体系中均具有良好适应性。 在三种复合菌种对酸性红的脱色体系中，体系Ⅲ（菌丝球与海藻酸钠固定化的酵母细胞混合培养）的脱色效果可达98%以上。对该体系而言，3种粒径固定化酵母小球（Ø1.1mm、1.8mm、3.1mm）中Ø1.8mm、3.1mm小球脱色率较为理想，而酵母细胞初始密度对脱色效果的影响不大。 KM3、YR-1分别具有处理刚果红废水和酸性红废水的应用前景，而KM3与YR-1联合应用对酸性红的脱色效果更好。

硅酸盐细菌的优选和发酵培养基的改进

硅酸盐细菌具有分解含钾矿物释放速效钾和产生植物生长刺激物质的作用，以硅酸盐细菌芽孢制成的硅酸盐细菌菌剂是生物肥料的一类重要产品.菌种性能、芽孢浓度和芽孢的存活能力是影响硅酸盐细菌菌剂质量的主要因素.该课题通过考察硅酸盐细菌菌株的适应性，辅以解钾活性综合评价了五株生产菌株的性能.研究探讨了芽孢存活能力与芽孢大小、芽孢浓度的关系，芽孢大存活能力强，芽孢浓度低易获得较大的芽孢.优化了原生产培养基，芽孢浓度稳定提高了4倍以上，芽孢形成率达98%，芽孢大小适中，生长时间、芽孢浓度稳定提高了4倍以上，芽孢形成率达98%，芽孢大小适中，生长时间、芽孢形成时间、发酵液pH和粘度均达到生产要求.同时对K<'+>对硅酸盐细菌生长的影响进行了研究，结果表明低浓度的K<'+>（约1×10<'-3>mol/L）对硅酸盐细菌的生长是有利的.

Enhanced Thermal Shock Resistance of Ceramics through Biomimetically Inspired Nanofins

We propose here a new method to make ceramics insensitive to thermal shock up to their melting temperature. In this method the surface of ceramics was biomimetically roughened into nanofinned surface that creates a thin air layer enveloping the surface of the ceramics during quenching. This air layer increases the heat transfer resistance of the surface of the ceramics by about 10 000 times so that the strong thermal gradient and stresses produced by the steep temperature difference in thermal shock did not occur both on the actual surface and in the interior of the ceramics. This method effectively extends the applications of existing ceramics in the extreme thermal environments.

Interface delamination of the thermal barrier coating subjected to local heating

To investigate the possible failure modes of the thermal barrier coating (TBC) used to protect the scramjet combustion chamber, the local heating via laser beam irradiation was utilized to simulate the service condition of high thermal flux and high temperature gradient. Firstly, the experimental method and process were described and the typical fracture morphology of the TBC under test were provided. Then, the theoretical and finite element modeling were carried out to study the temperature, deformation and stresses of the specimen when the top ceramic coat was subjected to local heating, and to demonstrate the mechanism on the failure of the TBC. It is revealed that the interface delamination shall appear and ultimately lead to the failure of the TBC under such thermal loading of local quick heating. According to the outcome of this study, the driving force of the interface delamination is influenced greatly by the key structural parameters and performance matching. Moreover, by utilizing the rules of the effects of these parameters on the fracture driving force, there is some possibility for the designer to optimize the performances of the TBC.

绞股蓝离体培养中影响皂甙产量的几个因素研究

绞股蓝(Gynostemma pentaphyllum)系葫芦科绞股蓝属植物，药用价值广泛，但其野生资源日趋减少，绞股蓝主要药效成分为绞股 蓝皂甙。利用组织和细胞培养生产绞股蓝皂甙是合理开发利用和保护绞股蓝资源的可能途径之一。本文对绞股蓝组织培养中培养基 的蔗糖和激素的组成以及各种胁迫条件：渗透压、重金属离子、真菌诱导物等对皂苷产量的影响进行了初步研究。其中，渗透压、 重金属离子、真菌诱导物对绞股蓝愈伤组织皂甙产量的影响尚未见报道。1. 蔗糖对绞股蓝愈伤组织之生长影响显著，2,4-D对绞股 蓝愈伤组织皂甙含量、产量影响显著。增加蔗糖用量，减少2，4-D的用量可提高皂甙产量。2. Mn++ 用量的提高抑制绞股蓝愈伤组 织的生长，但可促进皂甙含量、产量的提高。Mn++用量提高至MS培养基的20-30倍时可使皂甙产量增加近一倍，而提高Cu++浓度的 作用不明显。3. 甘露醇用量增加抑制绞股蓝愈伤组织的生长，但可使皂甙含量、产量提高。0.680mol·l-1甘露醇可使皂甙产量提 高83%，而Nacl较大抑制愈伤组织的生长并使皂甙产量降低。4. 米曲霉粗提物对绞股蓝愈伤组织生长先略微促进，然后抑制，而根 霉粗提物则使愈伤组织生长受抑制；两者对皂甙含量、产量的作用相似：在较低浓度范围内升高，然后下降。米曲霉粗提物可提高 产量一倍，根霉粗提物可提高42%。这些结果为高产细胞系的筛选和生长、生产培养条件的优化积累了资料。在综述部分，对植物 细胞培养中组织和器官分化、细胞结构变化、生化水平的变化与次生物合成和积累的关系作了讨论。Gynostemma pentaphyllum blongs to Gynostemma, Cucurbitaccae. It has a wide medical use, but its wild resource is threatened by people's excessive use. Its effective medical components are gypenosides. For reasonable use and protect its resource, it is a possible way to product gypenosides by plant tissue and cell culture. This paper has a primary study on the components of sucrose and hormones and a variety of stress conditions: osmostic pressure, heavy metal ion, fungal elicitors in the medium for the calli culture. The effects of osmostic pressure, heavy metal ion and fungal elicitors on the calli of Gynostemma pentaphyllum have not been reported. 1. Sucrose had a significant effect on the growth of the calli, 2,4 D had notable effects on the gypenosides content and production of the calli. Increased the concentration of sucrose and decreased the concentration of 2,4 D improved the production of gypenosides. 2. Increased the concentration of Mn++ inhibited the growth of the calli, but improved the content and production of gypenosides. The optimum concentration was 20-30 times as MS medium which improved the production 100%. Increased the concentration of Cu++ had not a notable effect. 3. Increased the concentration of mannitol inhibited the growth of the calli, but improved the content and production of gypenosides. The optimum concentration was 0.680mol·l-1 which improved the production 83%. Nacl apparently inhibited the growth of the calli and decreased the production of gypenosides. 4. The crude preparation of Aspergillus oryzae inhibited the growth of the calli that in low concentration. The crude praparation of Rhizopus formosensis inhibited the growth of the calli throughout. Their effects on the content and production of gypenosides are alike, but the former is higher than the latter. On the optimum concentration, each crude preparation improved the production 100% (Aspergillus oryzae), 42%(Rhizopus formosensis). These results has accumulated some informantion on the select of high yield cell strains and choose the best culture conditons for the growth and gypenosides product of the calli. In the review, it is discussed that the differentiation on tissue-organal, cellular and biochemical levels related to the synthesis and accumulation of secondary metabolites in plant culuture.