Time course of foreign gene integration and expression in transgenic fish embryos

Using a nuclear transplantation approach, the integration and expression of the green fluorescent protein (GFP) gene in the embryogenesis of transgenic leach (Misgurnus anguillicaudatus Cantor) have been studied. The GFP gene expression is first observed at the gastrula stage, which is consistent with the initiation of cell differentiation of fish embryos. The time course of the foreign gene expression is correlated with the regulatory sequences. The expression efficiency also depends on the gene configuration: the expression of pre-integrating circular plasmid at early embryos is higher than that of the linear plasmid. The integration of the GFP gene is first detected at the blastula stage and lasts for quite a long period. When two types of different plasmids are co-injected into fertilized eggs, the behavior of their integration and expression is not identical.

岷江柏种子的形态和发芽特征与贮藏生理动态研究

岷江柏(Cupressus chenggiana S. Y. Hu)是我国川甘地区特有的珍稀濒危乔木，一般生长在干旱的河谷区，在涵养水源和保护水 土等方面起着重要的作用。本文选择4个岷江柏种群，采用了野外调查和室内实验相结合的研究方法，调查岷江柏种群结实状况， 分析种子和球果形态特征，阐明种子发芽的基本特征，研究岷江柏种子贮藏过程中几个生理指标的动态变化特点，目的是为岷江柏 种苗繁育、自然更新能力评估以及珍稀濒危机制分析提供理论依据。研究得出如下结论：1.岷江柏球果呈椭球形，长为1.5～ 2.2cm,宽为1.5～1.9cm，质量为1.7～4.2g，球果鳞片数量为8～11片，球果内种子数量一般在40～70粒。岷江柏种子为椭圆形，长 为3.58～4.02mm，宽为3.10～3.15mm，厚为0.96～1.11mm，千粒重为3.1～3.5g。岷江柏的结实率很低，并且有显著的地理差异和 大小年差异。2. 岷江柏种子发芽温度范围是5℃～30℃,其中种子的适宜发芽温度范围是10℃～25℃。种子最适发芽温度随着贮藏 时间的增加而变化。在适宜温度范围内，种子发芽周期为20d。温度对种子的发芽势和T50有显著影响，对种子发芽率没有显著影响 ；光照有利于种子发芽；岷江柏种子的发芽特征是岷江柏保护种子资源、防止物种濒危的一种环境适应，有助于岷江柏种子提高发 芽率和幼苗的存活率。岷江柏种子是一种耐贮藏的正常性种子，在短期贮藏过程中，贮藏温度和种子含水量对于种子生理指标和种 子发芽没有显著影响。3. 岷江柏种子在短期贮藏过程中，千粒重没有显著变化；含水量都经历了先下降，再稳定的过程；粗脂肪 含量和可溶性糖含量逐渐降低；可溶性蛋白含量和丙二醛含量逐渐增加；脯氨酸含量在贮藏1～7个月时变化差异不明显，但是贮藏 7～10个月后显著增加。岷江柏种子的各个生理指标之间的相关性差异不显著。4. 岷江柏球果和种子的形态特征存在显著的地理差 异。岷江柏种子的发芽能力的地理性差异不大，种群间差异不大。岷江柏种群的地理差异由种群特征、生境特征和气候特征共同决 定。5. 在岷江柏的人工繁育中，对于刚刚采集的种子，发芽温度在15℃～25℃比较适合，其中以25℃最佳；而对于短期贮藏(4～ 10个月)后的种子，发芽温度在10℃～25℃均可，以15℃～20℃为最佳。野外播种的最适时间为4～6月，6～9月的间歇性干旱和降 水波动可能是限制岷江柏自然更新的因素之一。在短期贮藏过程中，种子可以采用常规室温贮藏，可以节约成本。Cupressus chenggiana is a specific and endangered plant in Sichuan and Gansu provinces of China, and it usually grows in dry valley and plays an important role in water supply and soil and water conservation in the dry valley of alpine and canyon region of southwest China. The research selected four Cupressus chenggiana populations and used the methods of the field investigation and the lab experiments. The fruiting characters of Cupressus chenggiana populations, the morphological characters of seeds and cones, the germination characters of seeds and the store physiological dymatics of several factors of seeds have been studied in order to give some theoretical advices on the artificial propagation and the ability of natural regeneration and the endangered principle of Cupressus chenggiana in the paper. The main results may be clarified as follows: 1. The cones of Cupressus chenggiana are ellipsoidal, length ranged from 1.5 to 2.2cm, with ranged from 1.5 to 1.9 cm, weight ranged from 1.7 to 4.2g, the number of cone squama ranged from 8 to 11, and the seed number of per cone ranged from 40 to 70. The seeds of Cupressus chenggiana are elliptical, length ranged from 3.58 to 4.02 mm, width ranged from 3.10 to 3.15 mm, thickness ranged from 0.96 to 1.11 mm, and the weight of 1000 seeds ranged from 3.1 to 3.5g. The fruiting rate of Cupressus chenggiana is very low, and the fruiting period of Cupressus chenggiana has the geographical differences and the big or small year differences. 2. Seed germination temperature is between 5℃ and 30℃, while the suited temperature is between 10℃ and 25℃. The optimum temperature of seed germination will change as the store time of seeds changes logner. The cycle of seed germination can persist 20 days in the range of the suited temperature. The germination temperatures have significant influences on the germination potential and T50, but have no significant infuluences on the germination rate. The photoperiod is in favor of seed germination. The characters of Cupressus chenggiana seed germination represent a kind of environmental adaptability to protect the seed sources and endangered species, and it can give help to increase the germination rate of seeds and the livability of seedings. The seeds of Cupressus chenggiana are a kind of orthodox seeds that can endure the long time storage. In the short time storage, the store temperatures and the moisture contents of seeds have no significant infuluences on the physiological factors and the germination of seeds, but the store time has significant influences on the physiological factors of seeds. 3. In the short store course of Cupressus chenggianna seeds, the 1000 seed weight has no significant variation; The moisture content descends at the beginning of the storage, but has no significant variation later; The crude fat content and the soluble sugar content descend gradually; The soluble protein content and MDA content increase gradually; The praline content has no significant variation after 1～7 months storage, but increase significantly after 7～10 months storage. The correlations of different physiological factors are not significant. 4. The morphological characters of cones and seeds of four populations exist significant differences. The germination of Cupressus chenggiana seeds has no significant geographical variation. The geographical variation of Cupressus chenggiana populations can be ascribed to the population characters, climate and environment. 5. In the course of artificial propagation of Cupressus chenggiana, it is favored that the germination temperature of newly collected seeds is between 15℃ and 25℃, while the optimum temperature is 25℃. After the short storage ranged from 4 months to 10 months, it is favored that the germination temperature is between 10℃ and 25℃, while the optimum temperature is ranged from 15℃ to 20℃. The field sowing optimum time is between April and June, and the interval drought and fallrain fluctuation between July and September may be one of the reasons that restrict natural regeneration of Cupressus chenggiana. In the short storage, seeds can be stored in the condition of room temperature.

On the propagation rate of the chemical waves observed during the course of CO oxidation on a Ag/Pt(110) composite surface

We have analyzed the propagation rate of the chemical waves observed during the course of CO oxidation on a Ag/Pt(I 10) composite surface that were reported in our previous papers [Surf Interface Anal. 2001, 32, 179; J. Phys. Chem. B 2002, 106, 5645]. In all cases, the propagation rate v can be adequately fitted as v = v(0) + D-0/d, in which v(0) and D-0 are constants, and d is the distance between the reaction front of the chemical wave and the boundary from which the chemical wave originates. We propose that the surface species responsible for the formation of the chemical wave comes from two paths: the adsorption of molecules in the gas phase on the surface and the migration from the adjacent surface with different catalytic activity. v(0) corresponds to the contribution from the surface species due to the adsorption, and D-0/d to that of the surface species that migrates from the adjacent surface. The rate equation clearly suggests that the observed chemical wave results from the coupling between adjacent surfaces with different catalytic activities during the course of heterogeneous catalysis. These results, together with our previous reports, provide a good fundamental understanding of spillover, an important phenomenon in heterogeneous catalysis.

Three phases of dark-recovery course from photoinhibition resolved by the chlorophyll fluorescence analysis in soybean leaves under field conditions

The chlorophyll fluorescence in soybean leaves was observed by a portable fluorometer CF-1000 under field conditions. On clear days, F-0 increased while F, and F-v/F-m decreased gradually in the morning. At midday F-O reached its maximum while F-v and F-v/F-m reached their minimum. The reverse changes occurred in the afternoon. At dusk these parameters could return to levels near those at dawn. Following exposure to a strong sunlight for more than 3 h, the dark-recovery process displayed three phases: (1) slow increases in F-0, F-v and F-v/F-m within the first hour; (2) a faster decrease in F-0 and faster increases in F-v and F-v/F-m within subsequent two hours; (3) a slow decrease in F-0 and slow increases in F-v and F-v/F-m within the fourth hour. In comparison with darkness, weak irradiance had no stimulating effect on the recovery from photoinhibition. Hence the photoinhibition in soybean leaves is mainly the reflection of reversible inactivation of some photosystem 2 reaction centres, but not the result of D1 protein loss.

Olympic Robot Building Manual

The 1989 AI Lab Winter Olympics will take a slightly different twist from previous Olympiads. Although there will still be a dozen or so athletic competitions, the annual talent show finale will now be a display not of human talent, but of robot talent. Spurred on by the question, "Why aren't there more robots running around the AI Lab?", Olympic Robot Building is an attempt to teach everyone how to build a robot and get them started. Robot kits will be given out the last week of classes before the Christmas break and teams have until the Robot Talent Show, January 27th, to build a machine that intelligently connects perception to action. There is no constraint on what can be built; participants are free to pick their own problems and solution implementations. As Olympic Robot Building is purposefully a talent show, there is no particular obstacle course to be traversed or specific feat to be demonstrated. The hope is that this format will promote creativity, freedom and imagination. This manual provides a guide to overcoming all the practical problems in building things. What follows are tutorials on the components supplied in the kits: a microprocessor circuit "brain", a variety of sensors and motors, a mechanical building block system, a complete software development environment, some example robots and a few tips on debugging and prototyping. Parts given out in the kits can be used, ignored or supplemented, as the kits are designed primarily to overcome the intertia of getting started. If all goes well, then come February, there should be all kinds of new members running around the AI Lab!

Medicina Dentária Digital – Presente e Futuro

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária

The DSpace Course - Item Submission Workflows

This module will introduce the item submission workflows available in DSpace. Workflows allow submissions to be checked before entering the repository. Submissions may be checked for accuracy, in order to improve the metadata, or simply to decide if they are OK to be archived. The module will show the three workflow steps available in DSpace, along with details about adding, changing and removing them from the submission process of collections.

Beyond simple imaging with energetic beams – nanopatterning, correlative microscopy and statistical analysis of inclusions

Electron microscopy (EM) has advanced in an exponential way since the first transmission electron microscope (TEM) was built in the 1930’s. The urge to ‘see’ things is an essential part of human nature (talk of ‘seeing is believing’) and apart from scanning tunnel microscopes which give information about the surface, EM is the only imaging technology capable of really visualising atomic structures in depth down to single atoms. With the development of nanotechnology the demand to image and analyse small things has become even greater and electron microscopes have found their way from highly delicate and sophisticated research grade instruments to key-turn and even bench-top instruments for everyday use in every materials research lab on the planet. The semiconductor industry is as dependent on the use of EM as life sciences and pharmaceutical industry. With this generalisation of use for imaging, the need to deploy advanced uses of EM has become more and more apparent. The combination of several coinciding beams (electron, ion and even light) to create DualBeam or TripleBeam instruments for instance enhances the usefulness from pure imaging to manipulating on the nanoscale. And when it comes to the analytic power of EM with the many ways the highly energetic electrons and ions interact with the matter in the specimen there is a plethora of niches which evolved during the last two decades, specialising in every kind of analysis that can be thought of and combined with EM. In the course of this study the emphasis was placed on the application of these advanced analytical EM techniques in the context of multiscale and multimodal microscopy – multiscale meaning across length scales from micrometres or larger to nanometres, multimodal meaning numerous techniques applied to the same sample volume in a correlative manner. In order to demonstrate the breadth and potential of the multiscale and multimodal concept an integration of it was attempted in two areas: I) Biocompatible materials using polycrystalline stainless steel and II) Semiconductors using thin multiferroic films. I) The motivation to use stainless steel (316L medical grade) comes from the potential modulation of endothelial cell growth which can have a big impact on the improvement of cardio-vascular stents – which are mainly made of 316L – through nano-texturing of the stent surface by focused ion beam (FIB) lithography. Patterning with FIB has never been reported before in connection with stents and cell growth and in order to gain a better understanding of the beam-substrate interaction during patterning a correlative microscopy approach was used to illuminate the patterning process from many possible angles. Electron backscattering diffraction (EBSD) was used to analyse the crystallographic structure, FIB was used for the patterning and simultaneously visualising the crystal structure as part of the monitoring process, scanning electron microscopy (SEM) and atomic force microscopy (AFM) were employed to analyse the topography and the final step being 3D visualisation through serial FIB/SEM sectioning. II) The motivation for the use of thin multiferroic films stems from the ever-growing demand for increased data storage at lesser and lesser energy consumption. The Aurivillius phase material used in this study has a high potential in this area. Yet it is necessary to show clearly that the film is really multiferroic and no second phase inclusions are present even at very low concentrations – ~0.1vol% could already be problematic. Thus, in this study a technique was developed to analyse ultra-low density inclusions in thin multiferroic films down to concentrations of 0.01%. The goal achieved was a complete structural and compositional analysis of the films which required identification of second phase inclusions (through elemental analysis EDX(Energy Dispersive X-ray)), localise them (employing 72 hour EDX mapping in the SEM), isolate them for the TEM (using FIB) and give an upper confidence limit of 99.5% to the influence of the inclusions on the magnetic behaviour of the main phase (statistical analysis).

Developing a health and wellbeing platform in a living lab setting: An action design research study

The world’s population is rapidly aging, which affects healthcare budgets, resources, pensions and social security systems. Although most older adults prefer to live independently in their own home as long as possible, smart living solutions to support elderly people at home did not reach mass adoption, yet. To support people age-in-place a Living Lab is established in one of the metropolitan areas in the Netherlands. The main goal of the Living Lab is to develop an online health and wellbeing platform that matches service providers, caretakers and users and to implement that platform in one particular city district. In this paper we describe the narrative of the action design research process that will give researchers insight how to deal with complex multi-stakeholder design projects as well as cooperation issues to develop an artifact in a real-life setting.

Robust test method for time-course microarray experiments.

BACKGROUND: In a time-course microarray experiment, the expression level for each gene is observed across a number of time-points in order to characterize the temporal trajectories of the gene-expression profiles. For many of these experiments, the scientific aim is the identification of genes for which the trajectories depend on an experimental or phenotypic factor. There is an extensive recent body of literature on statistical methodology for addressing this analytical problem. Most of the existing methods are based on estimating the time-course trajectories using parametric or non-parametric mean regression methods. The sensitivity of these regression methods to outliers, an issue that is well documented in the statistical literature, should be of concern when analyzing microarray data. RESULTS: In this paper, we propose a robust testing method for identifying genes whose expression time profiles depend on a factor. Furthermore, we propose a multiple testing procedure to adjust for multiplicity. CONCLUSIONS: Through an extensive simulation study, we will illustrate the performance of our method. Finally, we will report the results from applying our method to a case study and discussing potential extensions.

The Frequency and Impact of Exposure to Potentially Traumatic Events Over the Life Course.

We examined the frequency and impact of exposure to potentially traumatic events among a nonclinical sample of older adults (n = 3,575), a population typically underrepresented in epidemiological research concerning the prevalence of traumatic events. Current PTSD symptom severity and the centrality of events to identity were assessed for events nominated as currently most distressing. Approximately 90% of participants experienced one or more potentially traumatic events. Events that occurred with greater frequency early in the life course were associated with more severe PTSD symptoms compared to events that occurred with greater frequency during later decades. Early life traumas, however, were not more central to identity. Results underscore the differential impact of traumatic events experienced throughout the life course. We conclude with suggestions for further research concerning mechanisms that promote the persistence of post-traumatic stress related to early life traumas and empirical evaluation of psychotherapeutic treatments for older adults with PTSD.

The bigger the better? or what we know and what we still need to learn about anthracycline dose per course, dose density and cumulative dose in the treatment of breast cancer.

Clinical Trial

Association of fever and severe clinical course in bronchiolitis

Little attention has been given to the relation between fever and the severity of bronchiolitis. Therefore, the relation between fever and the clinical course of 90 infants (59 boys, 31 girls) hospitalised during one season with bronchiolitis was studied prospectively. Fever (defined as a single recording > 38.0°C or two successive recording > 37.8°C) was present in 28 infants. These infants were older (mean age, 5.3 v 4.0 months), had a longer mean hospital stay (4.2 v2.7 days), and a more severe clinical course (71.0%v 29.0%) than those infants without fever. Radiological abnormalities (collapse/consolidation) were found in 60.7% of the febrile group compared with 14.8% of the afebrile infants. These results suggest that monitoring of body temperature is important in bronchiolitis and that fever is likely to be associated with a more severe clinical course and radiological abnormalities.

[Book review] Chance encounters: a first course in data analysis and inference

Book review of: Chance Encounters: A First Course in Data Analysis and Inference by Christopher J. Wild and George A.F. Seber 2000, John Wiley & Sons Inc. Hard-bound, xviii + 612 pp ISBN 0-471-32936-3