Developing an HIV-1 vaccine has been hampered by the inability of immunogens to induce broadly neutralizing antibodies (bnAbs) that protect against infection. Previously, we used knockin (KI) mice expressing a prototypical gp41-specific bnAb, 2F5, to demonstrate that immunological tolerance triggered by self-reactivity of the 2F5 H chain, impedes bnAb induction. Here, we generate KI models expressing H chains from two other HIV-1 Abs: 4E10 (another self-/polyreactive, α-gp41 bnAb) and 48d (an α-CD4 inducible, non-polyreactive Ab), and find a similar developmental blockade consistent with central B-cell deletion in 4E10, but not in 48d VH KI mice. Furthermore, in KI strains expressing the complete 2F5 and 4E10 Abs as BCRs, we find that residual splenic B-cells arrest at distinct developmental stages, yet exhibit uniformly low surface Ig densities, elevated basal activation, profoundly muted responses to BCR ligation, and when captured as hybridoma mAb lines, maintain their dual (gp41/lipid) affinities and capacities to neutralize HIV-1, establishing a key role for anergy in suppressing residual 2F5 or 4E10-expressing B-cells. Importantly, serum IgGs from naïve 2F5 and 4E10 KI strains selectively eliminate gp41 and lipid binding, respectively, suggesting B-cells expressing 2F5 or 4E10 as BCRs exhibit specificity for a distinct spectrum of host antigens, including selective interactions by 2F5 BCR+ B-cells (i.e., and not 4E10 BCR+ B-cells) with residues in self-antigen(s) that mimic its gp41 neutralization epitope.

This paper summarizes the recent advances in image processing methods for morphological cell analysis. The topic of morphological analysis has received much attention with the increasing demands in both bioinformatics and biomedical applications. Among many factors that affect the diagnosis of a disease, morphological cell analysis and statistics have made great contributions to results and effects for a doctor. Morphological cell analysis finds the cellar shape, cellar regularity, classification, statistics, diagnosis, and so forth. In the last 20 years, about 1000 publications have reported the use of morphological cell analysis in biomedical research. Relevant solutions encompass a rather wide application area, such as cell clumps segmentation, morphological characteristics extraction, 3D reconstruction, abnormal cells identification, and statistical analysis. These reports are summarized in this paper to enable easy referral to suitable methods for practical solutions. Representative contributions and future research trends are also addressed.

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Small interfering RNAs (siRNAs) direct cleavage of complementary target RNAs via an RNA-induced silencing complex (RISC) that contains Argonatute2 protein at its core. However, what happens after target cleavage remains unclear. Here we analyzed the cleavage reaction by Drosophila Argonaute2-RISC using single-molecule imaging and revealed a series of intermediate states in target recognition, cleavage, and product release. Our data suggest that, after cleavage, RISC generally releases the 5' cleavage fragment from the guide 3' supplementary region first and then the 3' fragment from the seed region, highlighting the reinforcement of the seed pairing in RISC. However, this order can be reversed by extreme stabilization of the 3' supplementary region or mismatches in the seed region. Therefore, the release order of the two cleavage fragments is influenced by the stability in each region, in contrast to the unidirectional base pairing propagation from the seed to the 3' supplementary region upon target recognition.

Data envelopment analysis (DEA) is a useful method to evaluate the relative efficiency of peer decision making units (DMUs). Based upon the definitions of supply chain efficiency, we investigate the efficiency game between two supply chain members. It is shown that there exist numerous Nash equilibriums efficiency plans for the supplier and the manufacturer with respect to their efficiency functions. A bargaining model is then proposed to analyze the supplier and manufacturer's decision process and to determine the best efficiency plan strategy. DEA efficiency for supply chain operations is studied for the central control and the decentralized control cases. The current study is illustrated with a numerical example.Keywords Supply chain . Data envelopment analysis (DEA) . Efficiency . Nash equilibrium . Bargaining model Data Envelopment Analysis (DEA) was developed by Charnes, Cooper and Rhodes (CCR, 1978) to evaluate the relative efficiency of peer decision making units (DMUs). DEA has been proven an effective tool for performance benchmarking when multiple performance measures exist and a priori information on the tradeoffs among these measures is completely available (see, e.g., Zhu, 2003). The standard DEA approach treats each DMU's operation (e.g., production process) as a blackbox where inputs are transformed into outputs. The current paper addresses the issue of supply chain efficiency under the context of DEA.There are studies on applying the standard DEA model, e.g., CCR model, to measure the performance of supply chain members. For example, Weber and Desai (1996) employ DEA to construct an index of relative supplier performance. Easton et al. (2002) suggest a DEA model to compare the purchasing efficiency of firms in the petroleum industry. Some Y. Chen ( ) College

Influenza A viruses (IAVs) rely on host factors to support their life cycle, as viral proteins hijack or interact with cellular proteins to execute their functions. Identification and understanding of these factors would increase our knowledge of the molecular mechanisms manipulated by the viruses. In this study, we searched for novel binding partners of the influenza A virus NS2 protein, the nuclear export protein responsible for overcoming host range restriction, by a yeast two-hybrid screening assay and glutathione S-transferase-pulldown and coimmunoprecipitation assays and identified AIMP2, a potent tumor suppressor that usually functions to regulate protein stability, as one of the major NS2-binding candidates. We found that the presence of NS2 protected AIMP2 from ubiquitin-mediated degradation in NS2-transfected cells and AIMP2 functioned as a positive regulator of IAV replication. Interestingly, AIMP2 had no significant effect on NS2 but enhanced the stability of the matrix protein M1. Further, we provide evidence that AIMP2 recruitment switches the modification of M1 from ubiquitination to SUMOylation, which occurs on the same attachment site (K242) on M1 and thereby promotes M1-mediated viral ribonucleoprotein complex nuclear export to increase viral replication. Collectively, our results reveal a new mechanism of AIMP2 mediation of influenza virus replication. IMPORTANCEAlthough the ubiquitination of M1 during IAV infection has been observed, the precise modification site and the molecular consequences of this modification remain obscure. Here, we demonstrate for the first time that ubiquitin and SUMO compete for the same lysine (K242) on M1 and the interaction of NS2 with AIMP2 facilitates the switch of the M1 modification from ubiquitination to SUMOylation, thus increasing viral replication. Influenza A virus (IAV) is a significant cause of morbidity and mortality in both humans and animal species owing to its ability to cause yearly epidemics and occasional pandemics (1-3). Like other viruses, IAV hijacks the host cellular machinery to support its life cycle. Thus, identification and characterization of the interactions between viral components and specific host factors would help provide an understanding of the mechanisms by which the viruses acquire human pandemic potential.IAV belongs to the Orthomyxoviridae family, and its genome consists of eight negative-sense RNA segments encoding up to 17 viral proteins (4-9). The viral RNA (vRNA) exists as a form of viral ribonucleoprotein complexes (vRNPs) containing vRNA, nucleoprotein (NP), and three viral polymerase proteins (PB1, PB2, and PA). Unlike most other RNA viruses, influenza virus transcribes and replicates its genome in the nucleus. Thus, after it enters a host cell, vRNPs enter the nucleus to complete transcription and replication (10). The newly synthesized vRNPs are exported from the nucleus for packaging into progeny virions (11). In this regard, efficient nuclear export of vRNPs is essential for productive infection.NS2, also kn...

Therapeutic, V.

Gene therapy provides great opportunities for treating diseases from genetic disorders to immune system diseases and cancer. Much work has focused on bioactive macromolecules and nanoparticles, which can interact with low-molecular-mass drugs or DNA molecules to form complexes, and have been used for local delivery of therapeutic factors for tissue regeneration or gene therapy. In this study, inorganic amorphous calcium carbonate (ACC) hybrid nanospheres functionalized with a small amount of Ca(II)-IP 6 compound (CaIP 6 ) were prepared on a large scale by a facile gas-diffusion method. The results of in vitro transfection experiments show that the obtained functional calcium carbonate/phosphate nanocomposite particles had higher transfection efficiency and lower cytotoxicity level than commercial Lipofectamine 2000. From these results alternative new vectors for gene delivery could be developed. In addition, functionalized inorganic nanocomposite particles are biocompatible and biodegradable, thus the as-prepared hybrid nanospheres are promising for biomedical applications as a safe biomaterial.