Morphine Suppresses IFN Signaling Pathway and Enhances AIDS Virus Infection

Background: Opioids exert a profound influence on immunomodulation and enhance HIV infection and replication. However, the mechanism(s) of their action remains to be determined. We thus investigated the impact of morphine on the intracellular innate antiviral immunity. Methodology/Principal Findings: Seven-day-cultured macrophages were infected with equal amounts of cell-free HIV Bal or SIV DeltaB670 for 2 h at 37uC after 24 h of treatment with or without morphine. Effect of morphine on HIV/SIV infection and replication was evaluated by HIV/SIV RT activity assay and indirect immunofluorescence for HIV p24 or SIV p28 antigen. The mRNA expression of cellular factors suppressed or induced by morphine treatment was analyzed by the real-time RT-PCR. We demonstrated that morphine treatment of human blood monocyte-derived macrophages significantly inhibited the expression of interferons (IFN-a, IFN-b and IFN-l) and IFN-inducible genes (APOBEC3C/3F/3G and 3H). The further experiments showed that morphine suppressed the expression of several key elements (RIG-I and IRF-7) in IFN signaling pathway. In addition, morphine treatment induced the expression of suppressor of cytokine signaling protein-1, 2, 3 (SOCS-1, 2, 3) and protein inhibitors of activated STAT-1, 3, X, Y (PIAS-1, 3, X, Y), the key negative regulators of IFN signaling pathway. Conclusions: These findings indicate that morphine impairs intracellular innate antiviral mechanism(s) in macrophages

Establishment of Cre-mediated HBV recombinant cccDNA (rcccDNA) cell line for cccDNA biology and antiviral screening assays

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA), existing in hepatocyte nuclei as a stable minichromosome, plays a central role in the life cycle of the virus and permits the persistence of infection. Despite being essential for HBV infection, little is known about the molecular mechanisms of cccDNA formation, regulation and degradation, and there is no therapeutic agents directly targeting cccDNA, fore mostly due to the lack of robust, reliable and quantifiable HBV cccDNA models. In this study, combined the Cre/loxP and sleeping beauty transposons system, we established HepG2-derived cell lines integrated with 2-60 copies of monomeric HBV genome flanked by loxP sites (HepG2-HBV/loxP). After Cre expression via adenoviral transduction, 3.3-kb recombinant cccDNA (rcccDNA) bearing a chimeric intron can be produced in the nuclei of these HepG2-HBV/loxP cells. The rcccDNA could be accurately quantified by quantitative PCR using specific primers and cccDNA pool generated in this model could be easily detected by Southern blotting using the digoxigenin probe system. We demonstrated that the rcccDNA was epigenetically organized as the natural minichromosome and served as the template supporting pgRNA transcription and viral replication. As the expression of HBV S antigen (HBsAg) is dependent on the newly generated cccDNA, HBsAg is the surrogate marker of cccDNA. Additionally, the efficacies of 3 classes of anti-HBV agents were evaluated in HepG2-HBV/loxP cells and antiviral activities with different mechanisms were confirmed. These data collectively suggested that HepG2-HBV/loxP cell system will be powerful platform for studying cccDNA related biological mechanisms and developing novel cccDNA targeting drugs.</p

Soybean-derived Bowman-Birk inhibitor inhibits neurotoxicity of LPS-activated macrophages

<p>Abstract</p> <p>Background</p> <p>Lipopolysaccharide (LPS), the major component of the outer membrane of gram-negative bacteria, can activate immune cells including macrophages. Activation of macrophages in the central nervous system (CNS) contributes to neuronal injury. Bowman-Birk inhibitor (BBI), a soybean-derived protease inhibitor, has anti-inflammatory properties. In this study, we examined whether BBI has the ability to inhibit LPS-mediated macrophage activation, reducing the release of pro-inflammatory cytokines and subsequent neurotoxicity in primary cortical neural cultures.</p> <p>Methods</p> <p>Mixed cortical neural cultures from rat were used as target cells for testing neurotoxicity induced by LPS-treated macrophage supernatant. Neuronal survival was measured using a cell-based ELISA method for expression of the neuronal marker MAP-2. Intracellular reactive oxygen species (ROS) production in macrophages was measured via 2', 7'-dichlorofluorescin diacetate (DCFH₂DA) oxidation. Cytokine expression was determined by quantitative real-time PCR.</p> <p>Results</p> <p>LPS treatment of macrophages induced expression of proinflammatory cytokines (IL-1β, IL-6 and TNF-α) and of ROS. In contrast, BBI pretreatment (1-100 μg/ml) of macrophages significantly inhibited LPS-mediated induction of these cytokines and ROS. Further, supernatant from BBI-pretreated and LPS-activated macrophage cultures was found to be less cytotoxic to neurons than that from non-BBI-pretreated and LPS-activated macrophage cultures. BBI, when directly added to the neuronal cultures (1-100 μg/ml), had no protective effect on neurons with or without LPS-activated macrophage supernatant treatment. In addition, BBI (100 μg/ml) had no effect on N-methyl-D-aspartic acid (NMDA)-mediated neurotoxicity.</p> <p>Conclusions</p> <p>These findings demonstrate that BBI, through its anti-inflammatory properties, protects neurons from neurotoxicity mediated by activated macrophages.</p

Assemble Them All: Physics-Based Planning for Generalizable Assembly by Disassembly

Assembly planning is the core of automating product assembly, maintenance, and recycling for modern industrial manufacturing. Despite its importance and long history of research, planning for mechanical assemblies when given the final assembled state remains a challenging problem. This is due to the complexity of dealing with arbitrary 3D shapes and the highly constrained motion required for real-world assemblies. In this work, we propose a novel method to efficiently plan physically plausible assembly motion and sequences for real-world assemblies. Our method leverages the assembly-by-disassembly principle and physics-based simulation to efficiently explore a reduced search space. To evaluate the generality of our method, we define a large-scale dataset consisting of thousands of physically valid industrial assemblies with a variety of assembly motions required. Our experiments on this new benchmark demonstrate we achieve a state-of-the-art success rate and the highest computational efficiency compared to other baseline algorithms. Our method also generalizes to rotational assemblies (e.g., screws and puzzles) and solves 80-part assemblies within several minutes.Comment: Accepted by SIGGRAPH Asia 2022. Project website: http://assembly.csail.mit.edu

Establishment of linkage phase, using Oxford Nanopore Technologies, for preimplantation genetic testing of Coffin-Lowry syndrome with a de novo RPS6KA3 mutation

Background: This study aimed to perform preimplantation genetic testing (PGT) for a female Coffin-Lowry Syndrome (CLS) patient with a de novo mutation (DNM) in RPS6KA3. It was challenging to establish the haplotype in this family because of the lack of information from affected family members. Hence, we explored a new and reliable strategy for the detection of the DNM in PGT, using Oxford Nanopore Technologies (ONT) and the MARSALA platform.Methods: We performed whole-exome sequencing (WES) on the proband and confirmed the pathogenic mutation by Sanger sequencing. The proband then underwent PGT to prevent the transmission of the pathogenic mutation to her offspring. We diverged from the conventional methods and used long-read sequencing (LRS) on the ONT platform to directly detect the mutation and nearby SNPs, for construction of the haplotype in the preclinical phase of PGT. In the clinical phase of embryo diagnosis, the MARSALA method was used to detect both the SNP-based haplotype and chromosome copy number variations (CNVs), in each blastocyst. Finally, a normal embryo was selected by comparison to the haplotype of the proband and transferred into the uterus. Sanger sequencing and karyotyping were performed by amniocentesis, at 17 weeks of gestation, to confirm the accuracy of PGT.Results: Using WES, we found the novel, heterozygous, pathogenic c.1496delG (p.Gly499Valfs*25) mutation of RPS6KA3 in the proband. The SNP-based haplotype that was linked to the pathogenic mutation site was successfully established in the proband, without the need for other family members to be tested with ONT. Eight blastocysts were biopsied to perform PGT and were assessed with a haplotype linkage analysis (30 SNP sites selected), to give results that were consistent with direct mutation detection using Sanger sequencing. The results of PGT showed that three of the eight blastocysts were normal, without the DNM. Moreover, the patient had a successful pregnancy, after transfer of a normal blastocyst into the uterus, and delivered a healthy baby.Conclusion: The ONT platform, combined with the MARSALA method, can be used to perform PGT for DNM patients without the need for other samples as a reference

Learning Dense Reward with Temporal Variant Self-Supervision

Rewards play an essential role in reinforcement learning. In contrast to rule-based game environments with well-defined reward functions, complex real-world robotic applications, such as contact-rich manipulation, lack explicit and informative descriptions that can directly be used as a reward. Previous effort has shown that it is possible to algorithmically extract dense rewards directly from multimodal observations. In this paper, we aim to extend this effort by proposing a more efficient and robust way of sampling and learning. In particular, our sampling approach utilizes temporal variance to simulate the fluctuating state and action distribution of a manipulation task. We then proposed a network architecture for self-supervised learning to better incorporate temporal information in latent representations. We tested our approach in two experimental setups, namely joint-assembly and door-opening. Preliminary results show that our approach is effective and efficient in learning dense rewards, and the learned rewards lead to faster convergence than baselines.Comment: 4 pages, 6 figures, accepted to ICRA 2022 RL for Contact-Rich Manipulation Worksho

Pollution Assessment and Source Apportionment of Soil Heavy Metals in a Coastal Industrial City, Zhejiang, Southeastern China

In this research, Ningbo City, a typical industrial city in southeastern China, was selected as the study area, and the concentrations of 12 heavy metals (Cd, Cr, Ni, Pb, Zn, Cu, Hg, As, Co, V, Se, and Mn) were measured at 248 sampling points. Pollution index methods were used to assess the status of soil heavy metal contamination, and the Positive Matrix Factorization (PMF) model and Unmix model were integrated to identify and apportion the sources of heavy metal contamination. The results indicated that nearly 70% of the study area was polluted by heavy metals, and that Ni, Cr, and Zn were the main enriched heavy metals. The five sources identified using the PMF model were a geological source, an atmospheric deposition source, a transportation emissions source, a mixed source of agriculture and industry, and a mixed source of geology and industry. The four sources identified using the Unmix model were a mixed source of geology, agriculture, and industry (14.27%); a transportation emissions source (4.76%); a geological source (14.7%); and a mixed source of geology and industry (66.28%). These results have practical significance, as they can help to carry out pollution source risk assessment and give priority to the management of pollution source control