172 resultados para Rapid tests


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BACKGROUND: Previously, tachyplesin gene (tac) has been successfully transferred into Undaria pinnatifida gametophytes using the method of microprojectile bombardment transformation. The objectives of this study were to compare and evaluate the performance of bubble-column and airlift bioreactors to determine a preferred configuration of bioreactor for vegetative propagation of transgenic U. pinnatifida gametophytes, and to then investigate the influence of light on vegetative propagation of these gametophytes, including incident light intensity, photoperiod and light quality to resolve the problems of rapid vegetative propagation within the selected bioreactor. RESULTS: Experimental results showed that final dry cell density in the airlift bioreactor was 12.7% higher than that in the bubble-column bioreactor under the optimal aeration rate of 1.2 L air min(-1) L-1 culture. And a maximum final dry cell density of 2830 mg L-1 was obtained within the airlift bioreactor using blue light at 40 mu mol m(-2) s(-1) with a light/dark cycle of 14/10 (h). Polymerase chain reaction (PCR) analysis indicated that genes (bar and tac) were not lost during rapid vegetative propagation within the airlift bioreactor. CONCLUSION: The airlift bioreactor was shown to be much more suitable for rapid vegetative propagation of transgenic U. pinnatifida gametophytes than the bubble-column bioreactor in the laboratory. The use of blue light allows improvement of vegetative propagation of transgenic U. pinnatifida gametophytes. (C) 2009 Society of Chemical Industry

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A human acidic fibroblast growth factor gene, hafgf, was successfully transferred into Laminaria japonica (kelp) gametophytes via microprojectile bombardment using the biolistic PDS-1000/He gene gun. Following phosphinothricin screening, PCR detection and Southern blot analysis, transgenic L. japonica gametophytes were cultivated in an illuminated bubble-column bioreactor to optimize growth conditions. A maximal final dry cell density of 1,695 mg l(-1) was obtained in a batch culture having an initial dry cell density of 129.75 mg l(-1). This was achieved using an aeration rate of 1.08 l air min(-1) l(-1) culture in a medium containing 1.5 mM inorganic nitrate and 0.15 mM phosphate. In addition, the relationship between different nitrogen sources and growth of transgenic gametophytes indicated that both urea and sodium nitrate were effective nitrogen sources for cell growth, while ammonium ions inhibited growth of these gametophytes.

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Dot enzyme-linked immunosorbent assay (dot-ELISA), indirect ELISA and Western blot were performed to detect the virulent protease secreted by Vibrio anguillarum which was isolated from the diseased left-eyed flounder, Paralichthys olivaceous. Sensitivity results showed that dot-ELISA is a more sensitive, rapid and simple technique for the protease detection. The minimal detectable amount of protease is about 7 pg in the dot-ELISA test, while 7.8 ng in the indirect ELISA and 6.25 ng in the Western blot respectively. Protease could be detected 2 h after incubation of V. anguillarum in the 2216E liquid medium but enzyme activity was very low at that period. From 6 to 12 h, the amount and enzyme activity of protease increased markedly and reached maximum at stationary phase. Analysis of serum samples periodically collected from the infected flounders showed that after 2 h of infection by V. anguillarum, the pathogenic bacteria could be detected in the blood of the infected flounders but no protease was found. It was 5 similar to 6 h after infection that the protease was detected in blood and then the amount increased as infection advanced. Quantitative detection of protease either incubation in the medium or from the blood of infected flounders could be accomplished in virtue of positive controls of quantificational protease standards ("marker") so that the alterations of protease secretion both in vitro and in vivo could be understood generally. In addition, the indirect ELISA and dot-ELISA were also performed to detect V. anguillarum cells. Results indicated that the sensitivity of indirect ELISA to bacteria cells is higher than that of the dot-ELISA, and that the minimal detectable amount is approximately 10(4) cell/mL in the indirect ELISA, while 10(5) cell/mL in the dot-ELISA.

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Batch cultivation for transgenic kelp gametophyte cells was investigated in an online controlled 5 L stirred-tank photo-bioreactor to rapidly optimize the process conditions by monitoring the rate of increase of pH. The transgenic kelp gametophytes with heterologous gene encoding hepatitis B surface antigen (HBsAg) could rapidly grow in the bioreactor. Optimal temperature and agitation rate for bioreactor cultivation of gametophytes were 15 degrees C and 200 rpm. Optimal incident light intensities depended on the initial cell densities. (c) 2006 Elsevier B.V. All fights reserved.

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The ciliate community in the Bohai Sea (China) was studied from 23 September to 7 October 1998. A hurricane struck the study area between the 2 grid station investigations; which were 6 d apart. The ciliate biomass decreased drastically after the hurricane. The surface ciliate biomass decreased from 0.2-12.3 to 0.02-2.8 mug C l(-1) and the water column ciliate biomass from 2-136 to 0.01-47 mg C m(-2). The ciliate biomass:chlorophyll ratios in the surface water were calculated to be 0.04 to 4.7 (1.41 on average) during the first grid investigation and 0.01 to 1.18 (0.26 on average) during the second grid investigation. Distinct patterns of temporal changes in ciliate abundances were found at different stations both before and after the hurricane.

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Using the LAMP method, a highly specific and sensitive detection system for genetically modified soybean (Roundup Ready) was designed. In this detection system, a set of four primers was designed by targeting the exogenous 35S epsps gene. Target DNA was amplified and visualized on agarose gel within 45 min under isothermal conditions at 65 degrees C. Without gel electrophoresis, the LAMP amplicon was visualized directly in the reaction tube by the addition of SYBR Green I for naked-eye inspection. The detection sensitivity of LAMP was 10-fold higher than the nested PCR established in our laboratory. Moreover, the LAMP method was much quicker, taking only 70 min, as compared with 300 min for nested PCR to complete the analysis of the GM soybean. Compared with traditional PCR approaches, the LAMP procedure is faster and more sensitive, and there is no need for a special PCR machine or electrophoresis equipment. Hence, this method can be a very useful tool for GMO detection and is particularly convenient for fast screening.

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We investigated the independent and combined effects of experimental warming and grazing on plant species diversity on the north-eastern Tibetan Plateau, a region highly vulnerable to ongoing climate and land use changes. Experimental warming caused a 26-36% decrease in species richness, a response that was generally dampened by experimental grazing. Higher species losses occurred at the drier sites where N was less available. Moreover, we observed an indirect effect of climate change on species richness as mediated by plant-plant interactions. Heat stress and warming-induced litter accumulation are potential explanations for the species' responses to experimental warming. This is the first reported experimental evidence that climate warming could cause dramatic declines in plant species diversity in high elevation ecosystems over short time frames and supports model predictions of species losses with anthropogenic climate change.

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为提高自治水下机器人在垂直面跨越坡型、台阶型障碍的能力,提出了一种基于模糊控制的垂直面避碰规划方法。该方法以避碰声纳的输出作为输入,垂直面的深度调节量作为输出,直接建立了从障碍感知到避碰行为的映射。半物理仿真实验表明,该方法有效提高了某型自治水下机器人垂直面操纵性能,具有反应迅速、稳定性好、便于工程实现的优点。

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The Xiao-wan Power Station is the second highest arch dam in the world under construction. The height of the dam is about 292m. Large-scale excavation in the dam foundation of Xiao-wan Power Station has brought intensive unloading phenomenon. We collected a large number of firsthand data on unloaded rock mass in dam foundation, which supplies a natural testing ground for researching unloaded rock mass after excavation. Detailed study was carried out on the parameters of unloaded rock mass in the dam foundation of Xiao-wan Power Station. The study is not only importance for the Xiao-wan Power Station, but also has important instruction significance to similar projects in the Southwest of China. In order to study the mechanical parameters of unloaded rock mass, large field and laboratory tests were carried out. The test results showed the size effect of the sample is obvious. The change of deformation modulus of rock is not obvious. However, the Poisson's ratio of rock is increased under unloaded condition, its value is even more than 0.5. The theoretical forecasted results is accordance to the field tests including sound wave data and deformation monitoring data, which shows the forecasted results were reasonable. The soften yield criterion was adopted in the thesis to study the characteristics of the brittle rock mass in order to simulate their brittle failures. Based on the study results above, the transform of the structural plane network model to the numerical one was carried out, which made it feasible to consider the influence of large amount joints on the mechanical characteristic of rock mass in the numerical analyses. Using a factor, the degree of the damage or strengthen of rock mass can be determined rapidly, which proposed a rapid and feasible method for the determination of the parameters of rock mass.

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Geophysical inversion is a theory that transforms the observation data into corresponding geophysical models. The goal of seismic inversion is not only wave velocity models, but also the fine structures and dynamic process of interior of the earth, expanding to more parameters such as density, aeolotropism, viscosity and so on. As is known to all, Inversion theory is divided to linear and non-linear inversion theories. In rencent 40 years linear inversion theory has formed into a complete and systematic theory and found extensive applications in practice. While there are still many urgent problems to be solved in non-linear inversion theory and practice. Based on wave equation, this dissertation has been mainly involved in the theoretical research of several non-linear inversion methods: waveform inversion, traveltime inversion and the joint inversion about two methods. The objective of gradient waveform inversion is to find a geologic model, thus synthetic seismograms generated by this geologic model are best fitted to observed seismograms. Contrasting with other inverse methods, waveform inversion uses all characteristics of waveform and has high resolution capacity. But waveform inversion is an interface by interface method. An artificial parameter limit should be provided in each inversion iteration. In addition, waveform information will tend to get stuck in local minima if the starting model is too far from the actual model. Based on velocity scanning in traditional seismic data processing, a layer-by-layer waveform inversion method is developed in this dissertation to deal with weaknesses of waveform inversion. Wave equation is used to calculate the traveltime and derivative (perturbation of traveltime with respect to velocity) in wave-equation traveltime inversion (WT). Unlike traditional ray-based travetime inversion, WT has many advantages. No ray tracing or traveltime picking and no high frequency assumption is necessary and good result can be got while starting model is far from real model. But, comparing with waveform inversion, WT has low resolution. Waveform inversion and WT have complementary advantages and similar algorithm, which proves that the joint inversion is a better inversion method. And another key point which this dissertation emphasizes is how to give fullest play to their complementary advantages on the premise of no increase of storage spaces and amount of calculation. Numerical tests are implemented to prove the feasibility of inversion methods mentioned above in this dissertation. Especially for gradient waveform inversion, field data are inversed. This field data are acquired by our group in Wali park and Shunyi district. Real data processing shows there are many problems for waveform inversion to deal with real data. The matching of synthetic seismograms with observed seismograms and noise cancellation are two primary problems. In conclusion, on the foundation of the former experiences, this dissertation has implemented waveform inversions on the basis of acoustic wave equation and elastic wave equation, traveltime inversion on the basis of acoustic wave equation and traditional combined waveform traveltime inversion. Besides the traditional analysis of inversion theory, there are two innovations: layer by layer inversion of seimic reflection data inversion and rapid method for acoustic wave-equation joint inversion.

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Based on field survey, laboratory testing and numerical modeling, engineering characteristics of undisturbed loess and the mechanism of long-runout loess landslides caused by underground water level rise, as well as the formation conditions and spatial distribution of landslides, are systematically studied and analyzed. Loess landslides at south Plateau of Jingyang County are mainly classified as flowslide, slide and fall. Flowslide is the main type characteristic of high velocity, long runout and multi-stages. The steep relief composed of loose structured loess-old aged soil serials and the rise of groundwater table are the predominant conditions for landslides in the study area. To study loess mechanic poperties and loess landslides mechanisims, isotropically and anisotropically consolidated undrained compression(ICU and ACU) tests and constant-deviator-drained compression (CQD) tests were carried out on undisturbed samples. The results of undrained compression tests performed at the in-situ stress level show that the soils are of consistently strain-softening in the stress-strain relations and cause high excess pore pressure. The steady-state line and the potential region of instability are obtained from ICU and ACU test results. A necessary condition for liquefaction is that the soil state initially lies in or is brought into the potential instability region. In addition, a strong strain-softening model is also formed. CQD tests demonstrate that the mobilized friction angle is far less than the steady-state angle and that the soil experiences undrained contractive failure suddenly at very small strains when its stress path during drained loading tries to cross the potential instability region,thus validates the proposed instability region. Based on the location of the region of potential instability and the stress state of slope soil, a method of static liquefaction analysis is proposed for loess landslides caused by rise in groundwater table. Compared with other liquefaction analysis methods, this method overcomes the limitations inherent in conventional slope stability method and undrained brittleness index method. Triaxial tests composed of constant water content (CW) and wetting tests at constant deviator stress are performed on undisturbed unsaturated samples. The stress-strain relation of CW tests takes on strain-hardening behavior; The results of wetting tests at constant deviator stress designed to study the mechanics of failure of unsaturated loess caused by an increase in the degree of saturation (wetting) shows that a contractive failure occurs in the undisturbed samples. On the basis of the above triaxial test results, the initiation of static liquefaction is presented for long-runout loess landslides caused by rise in groundwater table, that is, the loess slope soil gradually transfer from unsaturated to saturated state under the infiltration of irrigation. A contractive failure occurs in the local region at very small strain by increasing the pore-water pressure at constant deviator stresses under drained conditons. It is the contractive failrue resulting from rise of pore pressure that leads to high excess pore pressure in the neighbour soil which reduces shear resistance of soil. The neighbour soils also fail due to the rapid increase in pore-water pressure. Thus a connected failure surface is developed quickly and a flowslide occurs. Based on the saturated-unsaturated seepage theory, transient seepage is computed using the finite element method on loess slope under groundwater table rise. Pore-water pressure distribution for every time step after irrigation are obtained. The phreatic surface in the slope increases with the groundwater table. Pore-water pressure distribution within 8m above the phreatic surface changes very quickly,but the water content and pore water pressure in the region ranging from 8m above the phreatic surface up to ground surface is almost not affected and the matric suction usually is kept at 100~120 kPa. Based on the results of laboratory tests and seepage flow analysis, the development process of loess landslide is modeled considering groundwater table rise. The shearing plastic zone first occurs at the slope toe where the soil is soaked for long term during rise in groundwater table. As irrigation continues, the shearing plastic zone gradually extends to the interior soils, with the results that the tensile plastic zone occurs at the slope crown. As time goes on, both the shearing plastic zone and tensile plastic zone continue to extend. Then a connected plastic zone is formed and fowslide occurs. In comparision to laboratory test results, the results of numerical simulation quite well verify the presented mechanism of static liquefaction of long-runout loess landslides caused by rise in groundwater table.

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A full understanding of failure mechanism, critical hydrological condition, and process of mobilization and deposition of a landslide is essential for optimal design of stabilization measure and forecasting of landslide hazard. This requires a quantitative study of hydrological response of a slope to rainfall through field monitoring, laboratory test and numerical modelling. At 13:40 on September 18, 2002, a fill slope failed following a period of prolonged rain in Shenzhen, resulting in 5 fatalities and 31 injuries. The failed mass with a volume about 2.5×104m3 traveled about 140m on level ground. Field monitoring, laboratory test, theoretical analysis and numerical modelling were carried out to undestand the hydrological response and failure mechanism of this fill slope. This thesis mainly focuses on the following aspects: (1) The hydrological responses and failure processes of slopes under rainfall infiltration were reviewed. Firstly, the factors influencing on the hydrological responses of slopes were analysed. Secondly, the change of stress state of slope soil and modelling methods of slope failure under rainfall infiltration were reviewed. (2) The characteristics of the Yangbaodi landslide and associated rainfall triggering the failure were presented. The failure was characterized by shallow flowslide, due to an increase of ground water table caused by rainfall infiltration. (3) A fully automated instrumentation was carried out to monitor rainfall, and saturated – unsaturated hydrological response of the fill slope, using a raingauge, piezometers, tensiometers and moisture probes. A conceptual hydrogeological model was presented based on field monitoring and borehole data. Analysis of monitoring data showed that the high pore water pressure in fill slope was caused by upward flow of semiconfined groundwater in the moderately decomposed granite. (4) Laboratory and in-situ testing was performed to study the physical and mechanical properties of fills. Isotropically consolidated undrained compression tests and anisotropically consolidated constant shear stress tests were carried out to understand the failure mechanism of the fill slope. It is indicated that loosely compacted soil is of strain-softening behaviour under undrained conditions, accompanied with a rapid increase in excess pore water pressure. In anisotropically consolidated constant shear stress tests, a very small axial strain was required to induce the failure and the excess pore water pressure increased quickly at failure. This indicated that static liquefaction caused by rise in groundwater table due to rainfall infiltration occurred. (5) The hydraulic conductivity of the highly and moderately decomposed granite was estimated using monitering data of pore water pressure. A saturated – unsaturated flow was modeled to study the hydrological response of the fill slope using rainfall records. It was observed that the lagged failure was due to the geological conditions and the discrepancy of hydraulic conductivity of slope soils. The hydraulic conductivity of moderately decomposed granite is relatively higher than the other materials, resulting in a semiconfied groundwater flow in the moderately decomposed granite, and subsequent upward flow into the upper fill layer. When the ground water table in the fill layer was increased to the critical state, the fill slope failed. (6) Numerical exercises were conducted to replay the failure process of the fill slope, based on field monitoring, laboratory and in-situ testing. It was found that the fill slope was mobilized by a rapid transfer of the concentrated shear stress. The movement of failure mass was characterized by viscosity fluid with a gradual increase in velocity. The failure process, including mobilization and subsequent movement and deposition, was studied using numerical methods.

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Study of dynamical mechanism of hydrocarbon secondary migration is the key research project of China Petroleum and Chemical Corporation in the ninth "Five-Year Plan", and this research is the hot and difficult spot and frontline in the domain of reservoir forming study in recent years. It is a common recognition that the dynamical mechanism of hydrocarbon's secondary migration is the important factor to control the reservoir type, distribution and oil/gas abundance. Therefore, to study this mechanism and establish the modes of hydrocarbon's migration and accumulation in different reservoirs under different conditions are of great theoretical meaningfulness and practical value on both developing the theory and method of hydrocarbon migration/accumulation dynamics in terrestrial rift-subsidence lacustrine basins and guiding the exploration and production. A laboratory for physical simulation of hydrocarbon's secondary migration/accumulation mechanism has been build up. 12 types of physical simulation tests to determine the volume of oil/gas migration and accumulation within these 3 series of plentiful hydrocarbon sources, different hydrocarbon abundance and pore level have been carried out under the guide of multidisciplinary theories, applying various methods and techniques, and 24 migration/accumulation modes have been established. The innovative results and recognition are as follows: 1, The oil/gas migration and accumulation modes for sandstones of moderate, fine grain size and silt in these six paleo depositional environments of shallow lake, fluvial, lacustrine, fluvial-deltaic, turbidite-delta, and salty-semi salty lake have been established. A new view has been put forward that the oil/gas volumetric increment during their migration and accumulation in different porous media of different rocks has similar features and evolution history. 2. During oil/gas migration and accumulation in different grain-sized porous media or different reservoirs, all the volumetric increment had experienced three period of rapid increasing, balanced and slower increasing and limited increasing. The dynamical process of oil/gas secondary migration and accumulation has been expounded. 3 The two new concepts of "source supply abundance" and "source supply intensity" have been proposed for the first time, and the physical simulation for hydrocarbon's migration, accumulation and forming a reservoir has been realized. 4, Source supply abundance is the important factor to control the accumulated volume of oil phase in the porous media. It is impossible to accumulate large amount of hydrocarbon volume in an open boundary system when the source supply abundance is low, i.e. impossible to form reservoirs of high productivity. 5 The above 12 types of physical simulation tests indicated that enough energy (pressure) of the oil sources is the decisive factor to ensure hydrocarbon's entering, flowing and accumulating through porous media, and both oil and gas phase will accumulate into the favorable places nearest to the oil sources. 6 The theory, method and related techniques for physical simulation of hydrocarbon's secondary migration/accumulation mechanism have been formed and applied to the E&P of Shengtuo rollover anticline and Niuzhuang turbidite lithological reservoirs. 7 This study developed the theory and method of hydrocarbon migration/accumulation dynamics in terrestrial rift-subsidence lacustrine basins, and the benefits and social effect are remarkable.

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Kela-2 gas field in Tarim Basin is the main supply source for West-to-East Pipeline project, also the largest abnormally-pressured gas field discovered in China currently. The geological characterization, fine geological modeling and field development plan are all the world-class difficult problems. This work includes an integrated geological and gas reservoir engineering study using advanced technology and approaches, the scientific development plan of Kela-2 gas field as well as the optimizations of the drilling, production and surface schemes. Then, it's expected that the Kela-2 gas field can be developed high-efficiently.Kuche depression is one part of the thrust belt of the South Tianshan Mountains, Kela-2 field is located at the Kelasu structural zone in the north of Kuche depression. The field territory is heavily rugged with deeply cut gullies, complex geological underground structure, variable rock types, thrust structure development. Therefore, considerable efforts have been made to develop an integrated technique to acquire, process and interpret the seismic data in complicated mountain region. Consequently a set of seismic-related techniques in the complicated mountain region has been developed and successfully utilized to interpret the structure of Kela-2 gas field.The main reservoir depositional system of Kela 2 gas field is a platform - fan delta - braided river system. The reservoir rocks are medium-fine and extremely fine grained sandstones with high structure maturity and low composition maturity. The pore system structure is featured by medium-small pore, medium-fine throat and medium-low assortment. The reservoir of Kela-2 gas field is characteristic of medium porosity and medium permeability. The pay zone is very thick and its lateral distribution is stable with a good connection of sand body. The overpressure is caused mainly by the strongly tectonic squash activities, and other factors including the later rapid raise and compartment of the high-pressure fluid, the injection of high-pressure fluid into the reservoir.Based on the deliverability tests available, the average binomial deliverability equation is provided applicable for the overall field. The experimental results of rock stress-sensitive tests are employed to analyze the change trend of petrophysical properties against net confining stress, and establish the stress-based average deliverability equation. The results demonstrate the effect of rock deformation on the deliverability is limited to less than 5% in the early period of Kela-2 gas field, indicating the insignificant effect on deliverability of rock deformation.In terms of the well pattern comparisons and development planning optimizations, it is recommended that the producers should be located almost linearly along the structural axis. A total of 9 producers have a stable gas supply volume of 10.76 BCMPY for 17 years. For Kela-2 gas field the total construction investment is estimated at ¥7,697,690,000 RMB with the internal earning rate of 25.02% after taxation, the net present value of ¥7,420,160,000 RMB and the payback period of 5.66 years. The high profits of this field development project are much satisfactory.