Development of a positive-negative selection procedure for gene targeting in fish cells

The gene targeting technique is a powerful tool for analyzing functions of cloned genes and for generating transgenic animals with site-directed integration of foreign genes. In order to develop this technique in fish, positive-negative selection (PNS) and homologous recombination vectors were constructed, and their expression was examined in fish cells. A vector (pNK) for PNS consists of the neomycin resistance gene (neo) as a positive selectable marker gene and the herpes simplex virus (HSV) thymidine kinase (tk) gene as a negative selectable marker gene. Positive selection with geneticin (G418) of epithelioma papulosum of carp (EPC) cells transfected with linearized pNK vector yielded 350 colonies, while double selection of transfected EPC cells with G418 and gancyclovir (Gc) resulted in nearly complete cell death, demonstrating that the PNS procedure is effective in fish cells. Homologous recombination vectors consist of the Xiphophorus melanoma receptor kinase (X mrk(Y)) gene as homologous sequence in addition to the neo and tk genes. Conditions for homologous recombination vector transfection and drug selection were established. After verification of the feasibility of expression of homologous recombination vectors in EPC cells, the first gene targeting experiments were attempted in the Xiphophorus melanoma cell line, PSM. Positive-negative selection of the targeting vector-transfectants led to a low enrichment in this particular cell line. The reasons for the low enrichment in PSM cells were discussed. (C) 2002 Elsevier Science B.V. All rights reserved.

Retention of the developmental pluripotency in medaka embryonic stem cells after gene transfer and long-term drug selection for gene targeting in fish

Embryonic stem (ES) cells provide a unique tool for introducing random or targeted genetic alterations, because it is possible that the desired, but extremely rare recombinant genotypes can be screened by drug selection. ES cell-mediated transgenesis has so far been limited to the mouse. In the fish medaka (Oryzias latipes) several ES cell lines have been made available. Here we report the optimized conditions for gene transfer and drug selection in the medaka ES cell line MES1 as a prelude for gene targeting in fish. MES1 cells gave rise to a moderate to high transfection efficiency by the calcium phosphate co-precipitation (5%), commercial reagents Fugene (11%), GeneJuice (21%) and electroporation (>30%). Transient gene transfer and CAT reporter assay revealed that several enhancers/promoters and their combinations including CMV, RSV and ST (the SV40 virus early gene enhancer linked to the thymidine kinase promoter) were suitable regulatory sequences to drive transgene expression in the MES1 cells. We show that neo, hyg or pac conferred resistance to G418, hygromycin or puromycin for positive selection, while the HSV-tk generated sensitivity to ganciclovir for negative selection. The positive-negative selection procedure that is widely used for gene targeting in mouse ES cells was found to be effective also in MES1 cells. Importantly, we demonstrate that MES1 cells after gene transfer and long-term drug selection retained the developmental pluripotency, as they were able to undergo induced differentiation in vitro and to contribute to various tissues and organs during chimeric embryogenesis.

Subtropical dipole mode in the Southern Hemisphere: A global view

A global wavenumber-3 dipole SST mode is showed to exist in the Southern Hemisphere subtropical climate variability in austral summer. A positive (negative) phase of the mode is characterized by cool (warm) SST anomalies in the east and warm (cool) SST anomalies in the southwest of the south Indian, Pacific, and Atlantic Oceans, respectively. This coherent dipole structure is largely a response of ocean mixed layer to the atmospheric forcing characterized by migration and modulation of the subtropical high-pressures, in which the latent heat flux play a leading role through wind-induced evaporation, although ocean dynamics may also be crucial in forming SST anomalies attached to the continents. Exploratory analyses suggest that this mode is strongly damped by the negative heat flux feedback, with a persistence time about three months and no spectral peak at interannual to decadal time scales. As the subtropical dipole mode is linearly independent of ENSO and SAM, whether it represents an additional source of climate predictability should be further studied. Citation: Wang, F. (2010), Subtropical dipole mode in the Southern Hemisphere: A global view, Geophys. Res. Lett., 37, L10702, doi: 10.1029/2010GL042750.

IMPACTS OF THE TROPICAL PACIFIC COUPLED PROCESS ON THE INTERANNUAL VARIABILITY IN THE INDIAN OCEAN

The basic features of climatology and interannual variations of tropical Pacific and Indian Oceans were analyzed using a coupled general circulation model (CGCM), which was constituted with an intermediate 2.5-layer ocean model and atmosphere model ECHAM4. The CGCM well captures the spatial and temporal structure of the Pacific El Nino-Southern Oscillation (ENSO) and the variability features in the tropical Indian Ocean. The influence of Pacific air-sea coupled process on the Indian Ocean variability was investigated carefully by conducting numerical experiments. Results show that the occurrence frequency of positive/negative Indian Ocean Dipole (IOD) event will decrease/increase with the presence/absence of the coupled process in the Pacific Ocean. Further analysis demonstrated that the air-sea coupled process in the Pacific Ocean affects the IOD variability mainly by influencing the zonal gradient of thermocline via modulating the background sea surface wind.

空间及言语工作记忆任务的情绪效应：来自ERP/ fMRI的证据

In the present study, based on processing efficiency theory, we used the event-related potentials (ERP) and functional magnetic resonance image (fMRI) techniques to explore the underlying neutral mechanism of influences of negative emotion on three subsystems of working memory, phonological loop、 visuospatial sketh pad and the central executive. The modified DSMT (delayed matching-to-sample task) and n-back tasks were adopted and IAPS (International Affective Picture System) pictures were employed to induce the expected emotional state of subjects. The main results and conclusions obtained in the series of experiments are as the following: 1. In DSM tasks, we found P200 and P300 were reduced by negative emotion in both spatial and verbal tasks, however the increased negative slow wave were only observed in spatial tasks, not in verbal tasks. 2. In n-back tasks, the updating function of WM associated P300 was affected by negative emotion only in spatial tasks, not in verbal tasks. Current density analysis revealed strong current density in the fronto-parietal cortex only in the spatial tasks as well. 3. We adopted fMRI-block design and ROIs analysis, and found significant emotion and task effects in spatial WM-associated right superior parietal cortex; only emotion effect in verbal WM-associated Broca’s area; the interaction effect in attention-associated medial prefrontal area and bilateral inferior parietal cortex. These results implied the negative emotion mainly disturbed the spatial WM-related areas, and the attention control system play a key role in the interaction of spatial WM and negative emotion. 4. to further examine the effects of positive、negative and neutral emotion on tasks with different cognitive loads, the selective effect of emotion on the ERP components of spatial WM was only found in 2-back tasks, not in visual searching tasks. So, firstly the positive emotion as well as negative emotion selectively disturbed on spatial WM in light of the attention resource competition mechanism. Secondly, the selective influences based on the different WM systems, not the properties of spatial and verbal information. At last, the manner of the interaction of emotion and cognition is correlated with the cognitive load.

老化态度对老年人记忆作业影响的研究

It was a study of the effect of non-cognitive factors on cognitive aging. The purpose of the study was to research the effect of attitude towards aging on memory performance of the elderly, and trying to explore the mechanism of the attitude towards aging on memory performance. The entire study was divided into three phases and included six experiments. The questionnaire of attitude towards aging was compiled in phase 1. Then the validity and reliability of it was tested in the elderly. The self-esteem scale was also tested as above. These prepare the instruments for the study. It was a static study in phase 2. Through comparing the old groups in different environment, analyzing relationship between different non-cognitive factors and memory performance in the old subjects in identical environment, the paths of the effect of the attitude towards aging on memory performance was explored. It was a dynamic study in phase 3. By changing attitude towards aging of the elderly with direct thinking (positive, negative), the effect on memory performance was observed and the mechanism was explored. Main results: 1. The validity and reliability of the questionnaire towards aging were good. Its construct had two factors (positive, negative). Both reliability of retest and split test were over 0.90. 2. Direct thinking task was able to change the attitude towards aging effectively in old or young adults. The scores of the attitudes towards aging between positive and negative thinking subjects were different significantly. 3. The effect of attitude towards aging on memory performance was significant. The results in static or dynamic condition showed that the elderly with higher score of the questionnaire of attitude towards aging (more positive), their memory self-efficacy and memory performance were higher. In the opposite (negative), they were lower. 4. Mechanism of the effect of the attitude towards aging on memory performance: in static condition there were two paths i.e. direct path and through memory self-efficacy; in dynamic condition, besides the two paths, emotion change constituted another path. 5. Different non-cognitive factors have different effect on psychological function. The results of this study found that self-esteem has no significant correlation with memory performance, but correlates with depression; The factor of education has effect on many non-cognitive factors and memory performance.

Phase control of switching from positive to negative index material in a four-level atomic system

Electric and magnetic responses of the medium to the probe field are analysed in a four-level loop atomic system by taking into account the relative phase of the applied fields. An interesting phenomenon is found: under suitable conditions, a change of the refractive index from positive to negative can occur by modulating the relative phase of the applied fields. Then the medium can be switched from a positive index material to a negative index material in our scheme. In addition, a negative index material can be realized in different frequency regions by adjusting the relative phase. It may give us a convenient way to obtain the desired material with positive or negative index.

Anomalous behaviour of photorefractive grating erasure owing to existence of both negative and positive carriers

In LiNbO3:Fe, anomalous behaviour of grating erasure is observed with different wavelenghts, i.e. rapid grating erasure in the short wavelength range, which deviates from the results predicted by the electron transport band model. The deviation is related to the coexistance of electrons and holes in photorefraction, and charge-transfer process including electrons and hole has been proposed. The electron and hole contributions to photo-excitation coefficient S of the Fe centre on the wavelength.

Large positive and negative lateral displacements from total internal reflection configuration with a weakly absorbing dielectric film

It is theoretically shown that the simultaneously large positive and negative lateral displacements will appear when the resonant condition is satisfied for a TE-polarized light beam reflected from the total internal reflection configuration with a weakly absorbing dielectric film. Appearance of the enhanced negative lateral displacement is relative to the incidence angle, absorption of the thin Elm and its thickness. If we select an appropriate weakly absorbing dielectric film and its thickness, the simultaneously enhanced positive and negative lateral displacements will appear at different resonant angles. These phenomena may lead to convenient measurements and interesting applications in optical devices.

Controllable negative and positive group delay in transmission through a single quantum well at finite magnetic fields

We investigate the controllable negative and positive group delay in transmission through a single quantum well at the finite longitudinal magnetic fields. It is shown that the magneto-coupling effect between the longitudinal motion component and the transverse Landau orbits plays an important role in the group delay. The group delay depends not only on the width of potential well and the incident energy, but also on the magnetic-field strengthen and the Landau quantum number. The results show that the group delay can be changed from positive to negative by the modulation of the magnetic field. These interesting phenomena may lead to the tunable quantum mechanical delay line. (c) 2007 Elsevier B.V. All rights reserved.

Studies on alkaloids binding to GC-rich human survivin promoter DNA using positive and negative ion electrospray ionization mass spectrometry

Electrospray ionization mass spectrometry (ESI-MS) was used to investigate the binding of 13 alkaloids to two GC-rich DNA duplexes which are critical sequences in human survivin promoter. Negative ion ESI-MS was first applied to screen the binding of the alkaloids to the duplexes. Six alkaloids (including berberine, jatrorrhizine, palmatine, reserpine, berbamine, and tetrandrine) show complexation with the target DNA sequences. Relative binding affinities were estimated from the negative ion ESI data, and the alkaloids show a binding preference to the duplex with higher GC content. Positive ion ESI mass spectra of the complexes were also recorded and compared with those obtained in negative ion mode.

Positive- and negative-ion matrix-assisted laser desorption/ionization mass spectrometry of saccharides

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) of oligosaccharides and polysaccharides has been investigated in detail, It is demonstrated that cationized species of oligosaccharides, [M+Na](+) and [M+K](+), are dominant products under the MALDI condition, and negative ions of oligosaccharides are not formed to any significant extent in this process, The molecular masses of polysaccharides are similarly determined by positive- and negative-ion MALDI-MS with the help of column chromatography. The distinction between positive- and negative-ion MALDI mass spectra of oligo-and polysaccharides indicates that the MALDI processes for saccharides vary with molecular mass. The matrix plays a more important role in the ionization process for oligosaccharides, while in the desorption process for polysaccharides. (C) 1998 John Wiley & Sons, Ltd.

The tunable negative refractive index in granular composite

The effective refractive index of a kind of granular composite, which consists of granular metallic and magnetic inclusions with different radius embedded in a host medium, is theoretically investigated. Results show that for certain volume fractions of these two inclusions, the negative permittivity peak shifts to low frequency and the peak value increases with increasing radius ratio of the radius of magnetic granulae to that of metallic granulae. Simultaneously, peak value of permeability decreases with the radius ratio, and value peak shifts to high frequency with increasing volume fraction of magnetic inclusion. Therefore, the radius ratio can affect the effective refractive index considerably, and it is found that by adjusting the radius ratio, the refractive index may change between negative and positive values for certain volume fractions of the two inclusions. (c) 2005 Elsevier B.V. All rights reserved.