We tested the ability of inactivated SARS-CoV vaccine to induce neutralizing antibodies in BALB/c mice. The inactivated vaccine was prepared by SARS-CoV virus propagation in Vero cells, with subsequent beta-propiolactone inactivation and Sepharose 4FF column chromatography purification. One hundred forty BALB/c female mice were divided into seven groups of 20 mice each. Of the seven groups, three groups were inoculated with 0.1, 1, and 3 microg of the vaccine without adjuvant while three other groups were inoculated at the same three dosages of vaccine with aluminum hydroxide as adjuvant, respectively. The remaining group was set up as a blank control. Each mouse was inoculated twice at an interval of 3 weeks. One week after the second immunization, mice sera were collected to detect serum neutralizing antibodies. An assay for determining neutralizing antibody titers was developed. The results can be summarized as follows: (1) higher dosages of vaccine induced higher levels of neutralizing antibody titer; (2) the level of neutralizing antibodies induced by the inoculation with aluminum hydroxide adjuvant was slightly higher than that without adjuvant, but the difference was not statistically significant.
The purified inactivated SARS vaccine could induce high levels of neutralizing antibody, and protect the monkeys from the challenge of SARS-CoV. The SARS vaccine prepared in the study appeared to be safe in monkeys.
The genome of Thermoanaerobacter tengcongensis is estimated to encode 2588 theoretical proteins. In this study, we have vitalized approximately 46% of the theoretical proteome experimentally using a proteomic strategy that combines three different methods, shotgun digestion plus high-performance liquid chromatography (HPLC) with ion-trap tandem mass spectrometry (shotgun-liquid chromatography (LC)/MS), one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) plus HPLC with ion-trap tandem mass spectrometry (one-dimensional electrophoresis (1DE)-LC/MS), and two-dimensional gel electrophoresis plus matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (2DE-MALDI-TOF-MS). Of the 1200 proteins identified, as few as 76 proteins were globally found by all three approaches, and notably, most of these proteins were in the soluble fraction. However, there were a number of unique proteins detected by one method only, suggesting that our strategy provides a means toward obtaining a comprehensive view of protein expression profile. Proteins from the major metabolic pathways are strongly represented on the map, and a number of these enzymes were identified by more than one proteomic method. Based upon the proteins identified in the present study, we are able to broaden the understanding of how T. tengcongensis survives under high temperature environment, whereas several of its properties can not be fully explained by genome data.
Mg2(Si,Sn)-based compounds have shown great promise for thermoelectric (TE) applications, as they are nontoxic and comprised abundantly available constituent elements. In this work, the crystal structures and TE properties of polycrystalline materials with nominal compositions Mg2Si0.35Sn0.65–x Bi x (x = 0, 0.015, 0.030, and 0.045) and Mg2Si y Sn0.97–y Bi0.03 (y = 0.30, 0.325, and 0.35) have been investigated. The electrical conductivity, Seebeck coefficient, and thermal conductivity are strongly affected by the presence of Bi. Undoped samples showed higher values of Seebeck coefficients (below 600 K), lower electrical conductivity, and lower thermal conductivity (above 600 K) in comparison to the Bi-doped samples. Furthermore, the signs of Seebeck coefficients are all negative, confirming that n-type conduction is dominant in these materials. Electrical conductivity was enhanced by increasing the Bi content up to 3% on the Si/Sn site because of the increasing amount of electron donors, and the absolute value of Seebeck coefficient decreased. When the Bi content is greater than 3%, lower zT values were obtained at 773 K. Thermal conductivity values might decrease with increasing Sn alloying for Mg2Si y Sn0.97–y Bi0.03, as mass and strain fluctuation caused by alloying can effectively scatter phonons. However, a different behavior was observed in higher Sn content material, possibly because of the absence of Mg atoms at the interstitial site [Mgi, on (1/2, 1/2, 1/2)] and vacancies of Mg atoms at the (1/4, 1/4, 1/4) site, as confirmed by Rietveld refinements. Outstanding figure of merit values in excess of unity were achieved with all samples, culminating in zT max = 1.35.
Filamentous fungi possess the capacity to produce a wide array of secondary metabolites with diverse biological activities and structures, such as lovastatin and swainsonine. With the advent of the post-genomic era, increasing amounts of cryptic or uncharacterized secondary metabolite biosynthetic gene clusters are continually being discovered. However, owing to the longstanding lack of versatile, comparatively simple, and highly efficient genetic manipulation techniques, the broader exploration of industrially important secondary metabolites has been hampered thus far. With the emergence of CRISPR/Cas9-based genome editing technology, this dilemma may be alleviated, as this advanced technique has revolutionized genetic research and enabled the exploitation and discovery of new bioactive compounds from filamentous fungi. In this review, we introduce the CRISPR/Cas9 system in detail and summarize the latest applications of CRISPR/Cas9-mediated genome editing in filamentous fungi. We also briefly introduce the specific applications of the CRISPR/Cas9 system and CRISPRa in the improvement of secondary metabolite contents and discovery of novel biologically active compounds in filamentous fungi, with specific examples noted. Additionally, we highlight and discuss some of the challenges and deficiencies of using the CRISPR/Cas9-based genome editing technology in research on the biosynthesis of secondary metabolites as well as future application of CRISPR/Cas9 strategy in filamentous fungi are highlighted and discussed.
BackgroundSeveral de novo transcriptome assemblers have been developed recently to assemble the short reads generated from the next-generation sequencing platforms and different strategies were employed for assembling transcriptomes of various eukaryotes without genome sequences. Though there are some comparisons among these de novo assembly tools for assembling transcriptomes of different eukaryotic organisms, there is no report about the relationship between assembly strategies and ploidies of the organisms.ResultsWhen we de novo assembled transcriptomes of sweet potato (hexaploid), Trametes gallica (a diploid fungus), Oryza meyeriana (a diploid wild rice), five assemblers, including Edena, Oases, Soaptrans, IDBA-tran and Trinity, were used in different strategies (Single-Assembler Single-Parameter, SASP; Single-Assembler Multiple-Parameters, SAMP; Combined De novo Transcriptome Assembly, CDTA, that is multiple assembler multiple parameter). It was found that CDTA strategy has the best performance compared with other two strategies for assembling transcriptome of the hexaploid sweet potato, whereas SAMP strategy with assembler Oases is better than other strategies for assembling transcriptomes of diploid fungus and the wild rice transcriptomes.ConclusionBased on the results from ours and others, it is suggested that CDTA strategy is better used for transcriptome assembly of polyploidy organisms and SAMP strategy of Oases is outperformed for those diploid organisms without genome sequences.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-014-1192-7) contains supplementary material, which is available to authorized users.
Electrically tunable evanescent mode half mode substrate integrated waveguide (HMSIW) resonators are implemented for S band applications. An HMSIW loaded with a complementary split ring resonator (CSRR) achieves forward electromagnetic wave transmission below the characteristic waveguide cutoff frequency due to evanescent wave amplification. A variable capacitor connected to one of the conductors of the CSRR changes its effective capacitance to ground, resulting in frequency tuning of the resonator. Three different configurations are investigated with a varactor diode connected between the ground and three different contact points of the CSRR. The external Q factor is slightly affected by the frequency tuning. More than 15% tunability is achieved around 3.4 GHz. Full wave structure simulation results are in good agreement with those of measurement.Index Terms-Complementary split ring resonator (CSRR), half mode substrate integrated waveguide (HMSIW), tunable resonator, varactor diode.
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