Novel epidithiodiketopiperazines as anti-viral zinc ejectors of the Feline Immunodeficiency Virus (FIV) nucleocapsid protein as a model for HIV infection

Focused libraries of multi-substituted epidithiodiketopiperazines (ETP) were prepared and evaluated for efficacy of inhibiting the nucleocapsid protein function of the Feline Immunodeficiency Virus (FIV) as a model for HIV. This activity was compared and contrasted to observed toxicity utilising an in-vitro cell culture approach. This resulted in the identification of several promising lead compounds with nanomolar potency in cells with low toxicity and a favorable therapeutic index.     

The comparison of genetic variation in the envelope protein between various immunodeficiency viruses and equine infectious anemia virus

The envelope protein (Env) of lentiviruses such as HIV, SIV, FIV and EIAV is larger than that of other retroviruses. The Chinese EIAV attenuated vaccine is based on Env and has helped to successfully control this virus, demonstrating that envelope is crucial for vaccine. We compared Env variation of the four kinds of lentiviruses. Phylogenetic analysis showed that the evolutionary relationship of Env between HIV and SIV was the closest and they appeared to descend from a common ancestor, and the relationship of HIV and EIAV was the furthest. EIAV had the shortest Env length and the least number of potential N-linked glycosylation sites (PNGS) as well as glycosylation density compared to various immunodeficiency viruses. However, HIV had the longest Env length and the most PNGS. Moreover, the alignment of HIV and SIV showed that PNGS were primarily distributed within extracellular membrane protein gp120 rather than transmembrane gp41. It implies that the size difference among these viruses is associated with a lentivirus specific function and also the diversity of env. There are low levels of modification of glycosylation sites of Env and selection of optimal protective epitopes might be useful for development of an effective vaccine against HIV/AIDS.     

Evaluation of the antiviral efficacy of bis[1,2]dithiolo[1,4]thiazines and bis[1,2]dithiolopyrrole derivatives against the nucelocapsid protein of the Feline Immunodeficiency Virus (FIV) as a model for HIV infection

A diverse library of bis[1,2]dithiolo[1,4]thiazines and bis[1,2]dithiolopyrrole derivatives were prepared for evaluation of activity against the nucleocapsid protein of the Feline Immunodeficiency Virus (FIV) as a model for HIV, using an in vitro cell culture approach, yielding nanomolar active compounds with low toxicity.     

FIV gp40七肽重复区基因的克隆表达及其产物的纯化

本研究利用Leam Coil-VMF程序预测到FIV Env蛋白gp40存在两个七肽重复区(Heptad repeat,HR1和HR2),对包括HR1和HR2在内的部分gp40胞外区基因(称为HRI-HR2)进行了人工合成,以此基因为模板获得了HR1和HR2基因的扩增产物,同时构建了用氨基酸连接子SGGRGG将HR1和HR2连接起来的串联基因(即HRIlinkerHR2,命名为2-Helix),采用人肠杆菌GST融合表达系统对HR1、HR2和2-Helix蛋白进行了表达,并对2-Helix进行了纯化。同时利用凝胶过滤层析证明2-Helix在PBS缓冲系统中以寡聚体的形式存在。     

FIV gp40 七肽重复区基因的克隆表达及其产物的纯化

本研究利用Learn Coil-VMF程序预测到FIV Env蛋白gp40存在两个七肽重复区(Heptad repeat,HR1和HR2),对包括HR1和HR2在内的部分gp40胞外区基因(称为HR1-HR2)进行了人工合成,以此基因为模板获得了HR1和HR2基因的扩增产物,同时构建了用氨基酸连接子SGGRGG将HR1和HR2连接起来的串联基因(即HR1linkerHR2,命名为2-Helix),采用大肠杆菌GST融合表达系统对HR1、HR2和2-Helix蛋白进行了表达,并对2-Helix进行了纯化.同时利用凝胶过滤层析证明2-Helix在PBS缓冲系统中以寡聚体的形式存在.     

On the role of long incubation periods in the dynamics of acquired immunodeficiency syndrome (AIDS)

In this study, we investigate systematically the role played by the reproductive number (the number of secondary infections generated by an infectious individual in a population of susceptibles) on single group populations models of the spread of HIV/AIDS. Our results for a single group model show that if R 1, the disease will die out, and strongly suggest that if R > 1 the disease will persist regardless of initial conditions. Our extensive (but incomplete) mathematical analysis and the numerical simulations of various research groups support the conclusion that the reproductive number R is a global bifurcation parameter. The bifurcation that takes place as R is varied is a transcritical bifurcation; in other words, when R crosses 1 there is a global transfer of stability from the infection-free state to the endemic equilibrium, and vice versa. These results do not depend on the distribution of times spent in the infectious categories (the survivorship functions). Furthermore, by keeping all the key statistics fixed, we can compare two extremes: exponential survivorship versus piecewise constant survivorship (individuals remain infectious for a fixed length of time). By choosing some realistic parameters we can see (at least in these cases) that the reproductive numbers corresponding to these two extreme cases do not differ significantly whenever the two distributions have the same mean. At any rate a formula is provided that allows us to estimate the role played by the survivorship function (and hence the incubation period) in the global dynamics of HIV. These results support the conclusion that single population models of this type are robust and hence are good building blocks for the construction of multiple group models. Our understanding of the dynamics of HIV in the context of mathematical models for multiple groups is critical to our understanding of the dynamics of HIV in a highly heterogeneous population.     

Analysis of recruitment and industrial human resources management for optimal productivity in the presence of the HIV/AIDS epidemic

The aim of this paper is to analyze the recruitment effects of susceptible and infected individuals in order to assess the productivity of an organizational labor force in the presence of HIV/AIDS with preventive and HAART treatment measures in enhancing the workforce output. We consider constant controls as well as time-dependent controls. In the constant control case, we calculate the basic reproduction number and investigate the existence and stability of equilibria. The model is found to exhibit backward and Hopf bifurcations, implying that for the disease to be eradicated, the basic reproductive number must be below a critical value of less than one. We also investigate, by calculating sensitivity indices, the sensitivity of the basic reproductive number to the model’s parameters. In the time-dependent control case, we use Pontryagin’s maximum principle to derive necessary conditions for the optimal control of the disease. Finally, numerical simulations are performed to illustrate the analytical results. The cost-effectiveness analysis results show that optimal efforts on recruitment (HIV screening of applicants, etc.) is not the most cost-effective strategy to enhance productivity in the organizational labor force. Hence, to enhance employees’ productivity, effective education programs and strict adherence to preventive measures should be promoted.     

Identification and characterization of nuclear localization signals within the nucleocapsid protein VP15 of white spot syndrome virus

The nucleocapsid protein VP15 of white spot syndrome virus (WSSV) is a basic DNA-binding protein. Three canonical bipartite nuclear localization signals (NLSs), called NLS1 (aa 11-27), NLS2 (aa 33-49) and NLS3 (44-60), have been detected in this protein, using the ScanProsite computer program. To determine the nuclear localization sequence of VP15, the full-length open reading frame, or the sequence of one of the three NLSs, was fused to the green fluorescent protein (GFP) gene, and transiently expressed in insect Sf9 cells. Transfection with full-length VP15 resulted in GFP fluorescence being distributed exclusively in the nucleus. NLS 1 alone could also direct GFP to the nucleus, but less efficiently. Neither of the other two NLSs (NLS2 and 3) was functional when expressed alone, but exhibited similar activity to NLS1 when they were expressed as a fusion peptide. Furthermore, a mutated VP15, in which the two basic amino acids (11RR12) of NLSI were changed to two alanines (11AA12), caused GFP to be localized only in the cytoplasm of Sf9 cells. These results demonstrated that VP15, as a nuclear localization protein, needs cooperation between its three NLSs, and that the two residues (11RR12) of NLS1 play a key role in transporting the protein to the nucleus.     

Guidelines for the prevention of simian immunodeficiency virus infection in laboratory workers and animal handlers

The authors are members of a working group that formulated guidelines to minimize transmission of simian immunodeficiency virus (SIV) infection to man. Biosafety level (BSL) 2 standards are recommended for handling of clinical specimens and housing of SIV inoculated animals. Manipulation of SIV preparations may be performed in a BSL 2 facility with additional BSL 3 practices and equipment; for large volume or concentrated preparations of SIV BSL 3 containment is necessary. Written policies regarding management and testing of workers exposed to SIV are recommended.     

The V1 region of gp120 is preferentially selected during SIV/HIV transmission and is indispensable for envelope function and virus infection

A transmission bottleneck occurs during each human immunodeficiency virus(HIV) transmission event, which allows only a few viruses to establish new infection. However, the genetic characteristics of the transmitted viruses that are preferentially selected have not been fully elucidated. Here, we analyzed amino acids changes in the envelope protein during simian immunodeficiency virus(SIV)/HIV deep transmission history and current HIV evolution within the last 15–20 years. Our results confirmed that the V1V2 region of gp120 protein, particularly V1, was preferentially selected. A shorter V1 region was preferred during transmission history, while during epidemic, HIV may evolve to an expanded V1 region gradually and thus escape immune recognition. We then constructed different HIV-1 V1 mutants using different HIV-1 subtypes to elucidate the role of the V1 region in envelope function. We found that the V1 region, although highly variable, was indispensable for virus entry and infection, probably because V1 deletion mutants exhibited impaired processing of gp160 into mature gp120 and gp41. Additionally, the V1 region affected Env incorporation. These results indicated that the V1 region played a critical role in HIV transmission and infection.     

A multiplexed protein microarray for the simultaneous serodiagnosis of human immunodeficiency virus/hepatitis C virus infection and typing of whole blood

All donor blood samples must be tested pretransfusion to determine the donor blood type. Standard testing protocols require that assays be performed for important bloodborne pathogens such as hepatitis C, syphilis, hepatitis B, and human immunodeficiency virus. We have demonstrated proof of the concept that a protein microarray can type whole blood and detect antibody to significant pathogens simultaneously from the same donor blood sample. The data collected demonstrate the ability of the array to accurately type blood samples while also detecting the presence of antibodies against both human immunodeficiency virus and hepatitis C virus. In conclusion, we have successfully developed a platform capable of typing human whole blood samples, while at the same time testing for the presence of antibodies specific for human immunodeficiency virus/hepatitis C virus. The major benefits of this system are its amenability to expansion with additional assays, for example, rhesus typing and syphilis and/or hepatitis B virus detection, and also the adaptability of the assay to higher-throughput analysis, currently 16 individual samples per slide, but readily expandable to a 96-well format.     

Serial expression of the truncated fragments of the nucleocapsid protein of CCHFV and identification of the epitope region

The Crimean-congo hemorrhagic fever virus (CCHFV) is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections. In this study, expression vectors carrying series truncated fragments of the NP (nucleocapsid protein) gene from the S fragment of CCHFV strain YL04057 were constructed. The recombinant proteins were expressed in E.coli and purified for detection. The antigenic of the truncated fragments of NP was detected with a polyclonal serum (rabbit) and 2 monoclonal (mAbs) (14B7 and 43E5) against CCHFV by Western-blot analyses. The results showed that the three expressed constructs, which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum. The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.     

吸毒人群中HIV/HCV核酸和抗体关系的分析

研究了高危人群中HIV/HCV核酸和抗体的关系。从新疆地区采集吸毒人群的血样,并对其进行HIV/ HCV核酸和抗体的检测。320例吸毒人员血浆样品中HCV抗体阳性为80．3％,HIV抗体阳性率为41．9％,HIV 和HCV共感染者为38．3％。HIV RNA与抗体的总符合率为98．8%,在186例HIV抗体阴性样品中可能有2例 为HIV感染的窗口期。HCV抗体和HCV RNA的阳性符合率为92．6％,HCV RNA与HCV抗体的总符合率为 90．0％,以上结果说明在HIV／HCV的高流行区进行HIV／HCV核酸检测可以发现病毒感染的窗口期,而约8% 的HCV抗体阳性样品为病毒核酸阴性,也值得进一步研究。     

Identification of unusual truncated forms of nucleocapsid protein in MDCK cells infected by Avian influenza virus (H9N2)

Unusual truncated forms of nucleocapsid protein (NP) were identified in the lysate of MDCK cells infected by Avian influenza virus (H9N2) using MS‐based proteomics approach. Moreover, O‐sulfonation that was considered as an unusual modification was identified in one of the tryptic peptides from the truncated NP. The findings might have implications on better understanding on the role of nucleoprotein in Avian influenza virus–host interaction.