Potential Application of the CRISPR/Cas9 System against Herpesvirus Infections.

The CRISPR/Cas9 system has been applied in the genome editing and disruption of latent infections for herpesviruses such as the herpes simplex virus, Epstein⁻Barr virus, cytomegalovirus, and Kaposi's sarcoma-associated herpesvirus. CRISPR/Cas9-directed mutagenesis can introduce similar types of mutations to the viral genome as can bacterial artificial chromosome recombination engineering, which maintains and reconstitutes the viral genome successfully. The cleavage mediated by CRISPR/Cas9 enables the manipulation of disease-associated viral strains with unprecedented efficiency and precision. Additionally, current therapies for herpesvirus productive and latent infections are limited in efficacy and cannot eradicate viruses. CRISPR/Cas9 is potentially adapted for antiviral treatment by specifically targeting viral genomes during latent infections. This review, which focuses on recently published progress, suggests that the CRISPR/Cas9 system is not only a useful tool for basic virology research, but also a promising strategy for the control and prevention of herpesvirus latent infections

Inhibition of Murine Cytomegalovirus Infection in Animals by RNase P-Associated External Guide Sequences.

External guide sequence (EGS) RNAs are associated with ribonuclease P (RNase P), a tRNA processing enzyme, and represent promising agents for gene-targeting applications as they can direct RNase-P-mediated cleavage of a target mRNA. Using murine cytomegalovirus (MCMV) as a model system, we examined the antiviral effects of an EGS variant, which was engineered using in vitro selection procedures. EGSs were used to target the shared mRNA region of MCMV capsid scaffolding protein (mCSP) and assemblin. In vitro, the EGS variant was 60 times more active in directing RNase P cleavage of the target mRNA than the EGS originating from a natural tRNA. In MCMV-infected cells, the variant reduced mCSP expression by 92% and inhibited viral growth by 8,000-fold. In MCMV-infected mice hydrodynamically transfected with EGS-expressing constructs, the EGS variant was more effective in reducing mCSP expression, decreasing viral production, and enhancing animal survival than the EGS originating from a natural tRNA. These results provide direct evidence that engineered EGS variants with higher targeting activity in vitro are also more effective in reducing gene expression in animals. Furthermore, our findings imply the possibility of engineering potent EGS variants for therapy of viral infections

RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins.

An engineered RNase P-based ribozyme variant, which was generated using the in vitro selection procedure, was used to target the overlapping mRNA region of two proteins essential for human cytomegalovirus (HCMV) replication: capsid assembly protein (AP) and protease (PR). In vitro studies showed that the generated variant, V718-A, cleaved the target AP mRNA sequence efficiently and its activity was about 60-fold higher than that of wild type ribozyme M1-A. Furthermore, we observed a reduction of 98%-99% in AP/PR expression and an inhibition of 50,000 fold in viral growth in cells with V718-A, while a 75% reduction in AP/PR expression and a 500-fold inhibition in viral growth was found in cells with M1-A. Examination of the antiviral effects of the generated ribozyme on the HCMV replication cycle suggested that viral DNA encapsidation was inhibited and as a consequence, viral capsid assembly was blocked when the expression of AP and PR was inhibited by the ribozyme. Thus, our study indicates that the generated ribozyme variant is highly effective in inhibiting HCMV gene expression and blocking viral replication, and suggests that engineered RNase P ribozyme can be potentially developed as a promising gene-targeting agent for anti-HCMV therapy

Atomic structures and deletion mutant reveal different capsid-binding patterns and functional significance of tegument protein pp150 in murine and human cytomegaloviruses with implications for therapeutic development.

Cytomegalovirus (CMV) infection causes birth defects and life-threatening complications in immunosuppressed patients. Lack of vaccine and need for more effective drugs have driven widespread ongoing therapeutic development efforts against human CMV (HCMV), mostly using murine CMV (MCMV) as the model system for preclinical animal tests. The recent publication (Yu et al., 2017, DOI: 10.1126/science.aam6892) of an atomic model for HCMV capsid with associated tegument protein pp150 has infused impetus for rational design of novel vaccines and drugs, but the absence of high-resolution structural data on MCMV remains a significant knowledge gap in such development efforts. Here, by cryoEM with sub-particle reconstruction method, we have obtained the first atomic structure of MCMV capsid with associated pp150. Surprisingly, the capsid-binding patterns of pp150 differ between HCMV and MCMV despite their highly similar capsid structures. In MCMV, pp150 is absent on triplex Tc and exists as a "Λ"-shaped dimer on other triplexes, leading to only 260 groups of two pp150 subunits per capsid in contrast to 320 groups of three pp150 subunits each in a "Δ"-shaped fortifying configuration. Many more amino acids contribute to pp150-pp150 interactions in MCMV than in HCMV, making MCMV pp150 dimer inflexible thus incompatible to instigate triplex Tc-binding as observed in HCMV. While pp150 is essential in HCMV, our pp150-deletion mutant of MCMV remained viable though with attenuated infectivity and exhibiting defects in retaining viral genome. These results thus invalidate targeting pp150, but lend support to targeting capsid proteins, when using MCMV as a model for HCMV pathogenesis and therapeutic studies

NGHIÊN CỨU XÂY DỰNG HỆ THỐNG HỖ TRỢ HÀNH CHÍNH TRỰC TUYẾN CHO SINH VIÊN TẠI TRƯỜNG ĐẠI HỌC NGOẠI NGỮ, ĐẠI HỌC HUẾ

Hệ thống hỗ trợ hành chính công trực tuyến là nơi tiếp nhận và xử lý đơn, xác nhận hành chính và các nhu cầu có liên quan của người học. Thời kỳ chuyển đổi số ngày nay đã tạo điều kiện thuận lợi để cải tiến công tác hành chính giúp người học đơn giản hóa thủ tục, đáp ứng nhanh các yêu cầu, số hóa một số quy trình và tư liệu hành chính của cơ sở giáo dục. Trong nghiên cứu này, chúng tôi xây dựng và thử nghiệm hệ thống hỗ trợ công tác hành chính công trực tuyến, qua đó khâu tiếp nhận yêu cầu từ người học và trả kết quả được tiến hành hoàn toàn trực tuyến. Hệ thống được triển khai trên giao diện web là trang Thông tin Đào tạo đại học của Trường Đại học Ngoại ngữ, Đại học Huế. Trên trang này, chúng tôi công khai các biểu mẫu hành chính, tiếp nhận các yêu cầu và xử lý, chuyển trả kết quả qua mạng. Bằng cách cho sinh viên sử dụng hệ thống và khảo sát người dùng để đánh giá, từ đó chúng tôi có căn cứ để đề xuất, điều chỉnh và đưa hệ thống vào ứng dụng thực tế với mục đích mang lại dịch vụ công tốt hơn cho người họ

Case Studies and Benchmark Examples for the Use of Grading Entropy in Geotechnics

The grading entropy concept can be adapted to the field of geotechnics, to establish criteria for phenomena such as particle packing, particle migration and filtering, through a quantified expression of the order/disorder in the grain size distribution, in terms of two entropy-based parameters. In this paper, the grading entropy theory is applied in some geotechnical case studies, which serve as benchmark examples to illustrate its application to the characterisation of piping, softening and dispersive soils, and to filtering problems in the context of a leachate collection system for a landfill site. Further, since unstable cohesive (dispersive) soils are generally improved by lime, the effect of lime addition is also considered, on the basis of some measurements and a further application of the grading entropy concept, which allows evolutions in the entropy of a soil to be considered as its grading is modified. The examples described support the hypothesis that the potential for soil erosion and particle migration can be reliably identified using grading entropy parameters derived from grading curve data, and applied through an established soil structure stability criteria and a filtering rule. It is shown that lime modification is not necessarily helpful in stabilizing against particle migration

Synthesis and Characterization of Silver Nanoplates by a Seed-mediated Method

Silver nanoplates (SNPs) having different size were synthesized by a seed-mediated method. The seeds -silver nanoparticles with 4 – 6 nm diameters were synthesized first by reducing silver nitrate with sodium borohydride in the present of Trisodium Citrate and Hydrogen peroxide. Then these seeds were developed by continue reducing Ag$^+$ ions with various amount of L-Ascorbic acid to form SNPs. Our analysis showed that the concentratrion of L-Ascorbic acid, a secondary reducing agent, played an important role to form SNPs. In addition, the size and in-plane dipole plasmon resonance wavelenght of silver nanoplates were increased when the concentration of added silver nitrate increased. The characterization of SNPs were studied by UV-Vis, FE-SEM, EDS and TEM methods

Some Notes on Granular Mixtures with Finite, Discrete Fractal Distribution

Why fractal distribution is so frequent? It is true that fractal dimension is always less than 3? Why fractal dimension of 2.5 to 2.9 seems to be steady-state or stable? Why the fractal distributions are the limit distributions of the degradation path? Is there an ultimate distribution? It is shown that the finite fractal grain size distributions occurring in the nature are identical to the optimal grading curves of the grading entropy theory and, the fractal dimension n varies between-¥ and ¥. It is shown that the fractal dimensions 2.2-2.9 may be situated in the transitional stability zone, verifying the internal stability criterion of the grading entropy theory. Micro computed tomography (μCT) images and DEM (distinct element method) studies are presented to show the link between stable microstructure and internal stability. On the other hand, it is shown that the optimal grading curves are mean position grading curves that can be used to represent all possible grading curves