Efficient RNA interference in zebrafish embryos using siRNA synthesized with SP6 RNA polymerase

Double-stranded RNA (dsRNA) has been shown to be a useful tool for silencing genes in zebrafish (Danio rerio), while the blocking specificity of dsRNA is still of major concern for application. It was reported that siRNA (small interfering RNA) prepared by endoribonuclease digestion (esiRNA) could efficiently silence endogenous gene expression in mammalian embryos. To test whether esiRNA could work in zebrafish, we utilized Escherichia coli RNaseIII to digest dsRNA of zebrafish no tail (ntl), a mesoderm determinant in zebrafish and found that esi-ntl could lead to developmental defects, however, the effective dose was so close to the toxic dose that esi-ntl often led to non-specific developmental defects. Consequently, we utilized SP6 RNA polymerase to produce si-ntl, siRNA designed against ntl, by in vitro transcription. By injecting in vitro synthesized si-ntl into zebrafish zygotes, we obtained specific phenocopies of reported mutants of ntl. We achieved up to a 59%no tail phenotype when the injection concentration was as high as 4 mu g/mu L. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization analysis showed that si-ntl could largely and specifically reduce mRNA levels of the ntl gene. As a result, our data indicate that esiRNA is unable to cause specific developmental defects in zebrafish, while siRNA should be an alternative for downregulation of specific gene expression in zebrafish in cases where RNAi techniques are applied to zebrafish reverse genetics.

Margulis' theory on division of labour in cells revisited

Division of labour is a marked feature of multicellular organisms. Margulis proposed that the ancestors of metazoans had only one microtubule organizing center (MTOC), so they could not move and divide simultaneously. Selection for simultaneous movement and cell division had driven the division of labour between cells. However, no evidence or explanation for this assumption was provided. Why could the unicellular ancetors not have multiple MTOCs? The gain and loss of three possible strategies are discussed. It was found that the advantage of one or two MTOC per cell is environment-dependent. Unicellular organisms with only one MTOC per cell are favored only in resource-limited environments without strong predatory pressure. If division of labour occurring in a bicellular organism just makes simultaneous movement and cell division possible, the possibility of its fixation by natural selection is very low because a somatic cell performing the function of an MTOC is obviously wasting resources. Evolutionary biologists should search for other selective forces for division of labour in cells.

Fabrication of a Low-Loss SSC Using High-Dose Electron Beam Lithography Exposure With Negative PMMA Resist

A silicon-on-insulator optical fiber-to-waveguide spot-size converter (SSC) using Poly-MethylMethAcrylate (PMMA) is presented for integrated optical circuits. Unlike the conventional use of PMMA as a positive resist, it has been successfully used as a negative resist with high-dose electron exposure for the fabrication of ultrafine silicon wire waveguides. Additionally, this process is able to reduce the side-wall roughness, and substantially depresses the unwanted propagation loss. Exploiting this technology, the authors demonstrated that the SSC can improve coupling efficiency by as much as over 2.5 dB per coupling facet, compared with that of SSC fabricated with PMMA as a positive resist with the same dimension.

Novel coupled multi-active region high power semiconductor lasers cascaded via tunnel junction

A novel semiconductor laser structure is put forward to resolve the major difficulties of high power laser diodes. In this structure, several active regions are cascaded by tunnel junctions to form a large optical cavity and to achieve super high efficiency. This structure can solve the problems of catastrophic optical damage of facet, thermal damage and poor light beam quality effectively. Low-pressure metalorganic chemical vapor deposition method is adopted to grow the novel semiconductor laser structures, which are composed of Si:GaAs/C:GaAs tunnel junctions, GaAs/InGaAs strain quantum well active regions. External differential quantum efficiency as high as 2.2 and light power output of 2.5 W per facet (under 2A drive current) are achieved from an uncoated novel laser device with three active regions.

Novel compact SOI-based reconfigurable optical add/drop multiplexer using microring resonators - art. no. 60190L

Novel compact design for 4-channel SOI-based reconfigurable optical add/drop multiplexer using microring resonators is presented and analyzed. Microring resonators have two important attributes as a key new technology for future optical communications, namely functionality and compactness. Functionality refers to the fact that a wide range of desirable filter characteristics can be synthesized by coupling multiple rings. Compactness refers the fact that ring resonators with radii about 30 mu m can lead to large scale integration of devices with densities on the order of 10(4) similar to 10(5) devices per square centimeter. A 4-channel reconfigurable optical add/drop multiplexer comprises a grid-like array of ridge waveguides which perpendicularly cross through each other. SOI-based resonators consisted of multiple rings at each of the cross-grid nodes serve as the wavelength selective switch, and they can switch an optical signal between two ports by means of tuning refractive index of one of the rings. The thermo-optic coefficient of silicon is 1.86x 10(-4) /K. Thus a temperature rise of 27K will increase the refractive index by 5 x 10(-3), which is enough to cause the switching of our designed microring resonators. The thermo-optic effect is used to suppress the resonator power transfer, rather than to promote loss. Thus, the input signal only suffers small attenuation and simultaneously low crosstalk can be achieved by using multiple rings.

Simulation of the Lanzhou Penning Trap LPT

The LPT (Lanzhou Penning Trap) is under construction and its task is to perforin direct mass measurement of fusion-evaporation residue, and if possible for heavy isotopes. Detailed simulations have been clone for a good understanding to the ion's movement and mechanics in the trap. The optimizal ion of the LPT is also performed based on the simulation. With a scale of 0.5 mm per grid used in the, simulation and many other limitations a highest mass resolution has been achieved to be 1.9 x 10(-5). An unexpected behavioin in the simulation related to magnetron motion has been found.

Novel Potentiometric Membrane Sensor for the Determination of Trace Amounts of Chromium(III) Ions

A plasticized Cr3+ ion sensor by incorporating 2,3,8,9-tetraphenyl-1,4,7,10-tetraazacyclododeca-1,3,7,9-tetraene (TTCT) ionophore exhibits a good potentiometric response for Cr3+ over a wide concentration range (1.0×10-6-1.0×10-1 M) with a slope of 19.5 mV per decade. The sensor response is stable for at least three months. Good selectivity for Cr3+ in comparison with alkali, alkaline earth, transition and heavy metal ions, and minimal interference are caused by Li+, Na+, K+, Co2+, Hg2+, Ca2+, Pb2+ and Zn2+ ions, which are known to interfere with other chromium membrane sensors. The TTCT-based electrode shows a fast response time (15 s), and can be used in aqueous solutions of pH 3 - 5.5. The proposed sensor was used for the potentiometric titration of Cr3+ with EDTA and for a direct potentiometric determination of Cr3+ content in environmental samples.

Characterization of stable isotope fractionation during methane production in the sediment of a eutrophic lake,lake dagow,germany

We measured delta C-13 of CO2, CH4, and acetate-methyl in profundal sediment of eutrophic Lake Dagow by incubation experiments in the presence and absence of methanogenic inhibitors chloroform, bromoethane sulfonate (BES), and methyl fluoride, which have different specificities. Methyl fluoride predominantly inhibits acetoclastic methanogenesis and affects hydrogenotrophic methanogenesis relatively little. Optimization of methyl fluoride concentrations resulted in complete inhibition of acetoclastic methanogenesis. Methane was then exclusively produced by hydrogenotrophic methanogenesis and thus allowed determination of the fractionation factors specific for this methanogenic pathway. Acetate, which was then no longer consumed, accumulated and allowed determination of the isotopic signatures of the fermentatively produced acetate. BES and chloroform also inhibited CH4 production and resulted in accumulation of acetate. The fractionation factor for hydrogenotrophic methanogenesis exhibited variability, e. g., it changed with sediment depth. The delta C-13 of the methyl group of the accumulated acetate was similar to the delta C-13 of sedimentary organic carbon, while that of the carboxyl group was by about 12 parts per thousand higher. However, the delta C-13 of the acetate was by about 5 parts per thousand lower in samples with uninhibited compared with inhibited acetoclastic methanogenesis, indicating unusual isotopic fractionation. The isotope data were used for calculation of the relative contribution of hydrogenotrophic vs. acetoclastic methanogenesis to total CH4 production. Contribution of hydrogenotrophic methanogenesis increased with sediment depth from about 35% to 60%, indicating that organic matter was only partially oxidized in deeper sediment layers.

Microarray-based Raman spectroscopic assay for kinase inhibition by gold nanoparticle probes

In this paper, a microarray-based surface-enhanced Raman spectroscopic (SERS) assay for detection of kinase functionality and inhibition has been reported. Biotinylated anti-phosphoserinen antibodies mark the phosphorylation and inhibition events and gold nanoparticles are attached to the antibodies by standard avidin-biotin chemistry, followed by silver deposition for SERS signal enhancement. The avidin conjugated fluorescein is used as SERS probe. The alpha-catalytic subunit of cyclic adenosine 5'-monophosphate (cAMP) dependent protein kinase (PKA), its well known substrate, kemptide, and three inhibitors, H89, HA1077, and KN62 have been chosen here to establish the SERS assay. As expected, highly selective inhibition of PKA is demonstrated with the inhibitor H89 and the inhibition assay enable to detect kinase inhibition as well as derive IC50 (half maximal inhibitory concentration) plots.

Microarray-based kinase inhibition assay by gold nanoparticle probes

We report on the development of a new class of kinase microarray for the detection of kinase inhibition based on marking peptide phosphorylation/biotinylation events by attachment of gold nanoparticles followed by silver deposition for signal enhancement. The alpha-catalytic subunit of cyclic adenosine 5'-monophosphate-dependent protein kinase (PKA), and its well-known substrate, kemptide, were used for the purpose of monitoring phosphorylation and inhibition. As expected, highly selective inhibition of PKA is demonstrated with the four inhibitors: H89, HA1077, mallotoxin, and KN62. Furthermore, an inhibition assay demonstrates the ability to detect kinase inhibition as well as derive IC50 (half-maximal inhibitory concentration) plots.

Cleavage of nucleotides by Ce4+ and the lanthanide metal complexes

The cleavage of adenosine-5'-monophosphate (5'-AMP) and guanosine-5'-monophosphate (S-GMP) by Ce4+ and lanthanide complex of 2-carboxyethylgermanium sesquioxide (Ge-132) in acidic and near neutral conditions was investigated by NMR, HPLC and measuring the liberated inorganic phosphate at 37 degrees C and 50 degrees C, The results showed that 5'-GMP and 5'-AMP was converted to guanine (G), 5'-monophosphate (depurination of 5'-GMP), ribose (depurination and dephosphorylation of 5'-GMP), phosphate and adenine (A), 5'-monophosphate (depurination of 5'-AMP), ribose (depurination and dephosphorylation of 5'-AMP), phosphate respectively by Ce4+. In presence of lanthanide complexes, 5'-GMP and 5'-AMP were converted to guanosine (Guo) and phosphate and adenosine (Ado) and phosphate respectively. The mechanism of cleaving 5'-GMP and 5'-AMP is hydrolytic scission.

Stability of Mo/Si multilayer structure used in Bragg-Fresnel optics

The thermal and chemical stabilities of Mo/Si multilayer structure used in Bragg-Fresnel optics were studied to get optimal technological parameters of pattern generation. Mo/Si multilayers were annealed at temperature ranging from 360 to 770 K, treated with acetone and 5 parts per thousand NaOH solution, and characterized by small-angle x-ray diffraction technique as well as x-ray photoelectron spectroscopy, and Olympus microscopy.

attB site disruption in marine Actinomyces sp M048 via DNA transformation of a site-specific integration vector

An efficient conjugation method has been developed for the marine Actinomyces sp. isolate M048 to facilitate the genetic manipulation of the chandrananimycin biosynthesis gene cluster. A phi C31-derived integration vector pIJ8600 containing oriT and attP fragments was introduced into strain M048 by bi-parental conjugation from Escherichia coli ET12567 to strain M048. Transformation efficiency was (6.38 +/- 0.41) x 10(-5) exconjugants per recipient spore. Analysis of eight exconjugants showed that the plasmid pIJ8600 was stably integrated at a single chromosomal site (attB) of the Actinomyces genome. The DNA sequence of the attB was cloned and shown to be conserved. The results of antimicrobial activity analysis indicated that the insertion of plasmid pIJ8600 seemed to affect the biosynthesis of antibiotics that could strongly inhibit the growth of E. coli and Mucor miehei (Tu284). HPLC-MS analysis of the extracts indicated that disruption of the attB site resulted in the complete abolition of chandrananimycin A-C production, proving the identity of the gene cluster. Instead of chandrananimycins, two bafilomycins were produced through disruption of the attB site from the chromosomal DNA of marine Actinomyces sp. M048.

东黄海海洋蓝细菌（Svnechococcus）生态学研究

该文对中国黄海和东海的蓝细菌在春、秋两季的生态分布特点进行了研究和比较,对黄海蓝细胞从10月至次年6月的生态分布进行了研究.1.东黄海海区：在秋季（2000年10月19日至11月29日）和春季（2001年3月26日至4月24日）两个季节中蓝细菌丰度在0.19～7.84×10<'4>cell/ml之间,生物量在0.56～13.74μgC/L之间,整体水平上春季比秋季高（蓝细菌的丰充与生物量平均值春季分析为4.83×10<'4>cell/ml和7.25μgC/L;秋季分别为1.72×10<'4>cell/ml和5.07μgC/L）,东海比黄海高（蓝细菌生物量平均值春季黄海和东海分别为4.42μgC/L和5.52μgC/L;秋季黄海和东海分别为7.02μgC/L和7.45μgC/L）.2.黄海海区：黄海蓝细菌丰度与生物量随时间变化趋势为10月至12月降低（蓝细菌丰度分别为15×10<'4>cell/ml和1.19×10<'4>cell/ml,生物量分别为4.41μgC/L和3.49×10<'4>cell/ml）,4月（蓝细菌丰度与生物量分别为4.68×10<'4>cell/ml和7.02μgC/L）与12月相比有明显增高,蓝细菌丰度4月是12月的3.93倍,生物量4月是12月的2.01倍,4月至6月蓝细菌生物量大致稳定,略有增加（5月和6月蓝细菌丰度分别为5.23×10<'4>cell/ml和5.25×10<'4>cell/ml,生物量分别为7.85μgC/L和7.88μgC/L）.研究发现蓝细菌在水体中的垂直分布与温度变化比较一致.

黄、东海海洋异养细菌的生态学研究

该文利用表面荧光显微镜观测计数法和[甲基-<'-3>H]胸腺嘧啶示踪法对春秋两季节中国黄、东海异养细菌生态分布及其生产力状况,以及异养细菌及其生产力与浮游植物叶绿素、无机氮盐之间的关系进行了研究.主要内容如下：春秋两季节黄东海海洋异养细菌丰度大致相当,异养细菌平均值在5～7×10<'8>Cell/l范围之间.春季异养细菌丰度总体表现为随离岸越远先逐渐降低后略微增高的趋势,秋季异养细菌丰度有离岸越远逐渐增高的趋势.秋季黄、东海海洋异养细菌生产力明显高于春季,并且秋季异养细菌的生长率也高于春季.秋季异养细菌生产力与初级生产的比值明显高于春季,且东海高于黄海.春季异养细菌生物量与浮游植物叶绿素及无机氮盐之间无明显相关性.秋季异养细菌生物量与浮游植物叶绿素相关性在东海有较显著相关性,在黄海该相关性比东海低;异养细菌生物量与NO<,3>ˉ和DIN表现出明显相关性,该相关性在黄海明显高于东海.异养细菌在一年的变化有明显规律,冬季最低,秋季和春季相当.在一天当中,异养细菌数量变化有一定规律性,且其变化与浮游植物细胞数量变化有一定关系.