Interfacial Engineering of Ru Nanocluster-Modified TiO2 Nanotube-Assisted Regulation of Lithium Polysulfide Reactions.

The inhibition of lithium polysulfide (LiPS) diffusion and the acceleration of reaction kinetics are two major challenges for the practical application of lithium-sulfur (Li-S) batteries. Herein, through an interface engineering strategy, a multifunctional sulfur host based on Ru nanocluster-modified TiO2 nanotubes (TiO2-Ru) was designed. The TiO2-Ru interface field effect, combined with the hollow nanotube structure and the strong chemical action of TiO2, enhanced the LiPS trapping ability and inhibited the "shuttle effect". Furthermore, the high catalytic activity of Ru nanoclusters reduced the energy barrier of multistep LiPS reactions, thus speeding up the electrode kinetics. As a result, the TiO2-Ru-based composite sulfur cathode delivered excellent electrochemical performance, including an extremely low capacity loss of ∼0.015% per cycle and an increased areal capacity of ∼6.1 mAh cm-2 at 4.8 mg cm-2. This work contributes to a better sulfur cathode design from insights into morphology and phase interface engineering.

A New HIV-1 K28E32-Reverse Transcriptase Variant Associated with the Rapid Expansion of CRF07_BC among Men Who Have Sex with Men.

HIV-1 CRF07_BC originated among injection drug users (IDUs) in China. After diffusing into men who have sex with men (MSM), CRF07_BC has shown a rapid expansion in this group; however, the mechanism remains unclear. Here, we identified a new K28E32 variant of CRF07_BC that was characterized by five specific mutations (E28K, K32E, E248V, K249Q, and T338S) in reverse transcriptase. This variant was mainly prevalent among MSM, and was overrepresented in transmission clusters, suggesting that it could have driven the rapid expansion of CRF07_BC in MSM, though founder effects cannot be ruled out. It was descended from an evolutionary intermediate accumulating four specific mutations and formed an independent phylogenetic node with an estimated origin time in 2003. The K28E32 variant was demonstrated to have significantly higher in vitro HIV-1 replication ability than the wild type. Mutations E28K and K32E play a critical role in the improvement of in vitro HIV-1 replication ability, reflected by improved reverse transcription activity. The results could allow public health officials to use this marker (especially E28K and K32E mutations in the reverse transcriptase (RT) coding region) to target prevention measures prioritizing MSM population and persons infected with this variant for test and treat initiatives. IMPORTANCE HIV-1 has very high mutation rate that is correlated with the survival and adaption of the virus. The variants with higher transmissibility may be more selective advantage than the strains with higher virulence. Several HIV-1 variants were previously demonstrated to be correlated with higher viral load and lower CD4 T cell count. Here, we first identified a new variant (the K28E32 variant) of HIV-1 CRF07_BC, described its origin and evolutionary dynamics, and demonstrated its higher in vitro HIV-1 replication ability than the wild type. We demonstrated that five RT mutations (especially E28K and K32E) significantly improve in vitro HIV-1 replication ability. The appearance of the new K28E32 variant was associated with the rapidly increasing prevalence of CRF07_BC among MSM.

Respiratory syncytial virus infection of microglia exacerbates SH-SY5Y neuronal cell injury by inducing the secretion of inflammatory cytokines: A Transwell in vitro study.

OBJECTIVES: To elucidate the mechanism of Respiratory Syncytial Virus (RSV) infection and central neuronal disease and to understand the role of microglia in neuronal injuries during RSV infection. MATERIALS AND METHODS: The effects of RSV and the cytokines produced by RSV-infected CHME-5 microglial cells on SY5Y neuronal cells were evaluated based on an in vitro Transwell coculture system. Five treatment groups were established in this study, including the normal control SY5Y group, RSV+SY5Y infection group, (cytokine+CHME-5)+SY5Y Transwell group, (RSV+CHME-5)+SY5Y Transwell group, and (RSV+cytokine+CHME-5)+SY5Y Transwell group. The morphological and physical alterations in SY5Y cells and their synapses were analyzed by confocal microscopy. The mRNA and protein expression levels of TLR3/RIG-I, as well as the expression of Hv1, in microglia were measured by qRT-PCR and Western blot assays. In addition, the apoptosis ratio of neuronal cells was determined by flow cytometry. RESULTS: RSV infection activated the protein expression of Hv1 protein in microglia in vitro (P<0.05), induced morphological changes in SY5Y cells, lengthened synapses (73.36±0.12 µm vs 38.10±0.11 µm), simultaneously activated TLR3 and RIG-I protein expression (P<0.05), upregulated the secretion of the inflammatory cytokines TNF-α, IL-6, and IL-8 (P<0.01), and increased the apoptosis rate of SY5Y cells (P<0.01). CONCLUSION: The results demonstrate that RSV infection of microglia can induce SY5Y neuronal cell injury and stimulate apoptosis through inflammatory cytokine release.

Near Full-Length Genomic Characterization of 16 HIV-1 CRF01_AE Primary Isolates from Guangxi, China.

Isolation and culture of human immunodeficiency virus (HIV) are an important basis for acquired immune deficiency syndrome (AIDS) etiology, immunology, drug screening, clinical treatment, and vaccine research. CRF01_AE is one of the predominant strains of HIV-1 in China. However, there are few HIV-1 CRF01_AE isolates that have been reported. In this study, 16 HIV-1 CRF01_AE strains from Guangxi, China, were isolated, and the near full-length genomes were reverse transcribed and amplified in two halves with the 1 kb overlapping region. The polymerase chain reaction products were sequenced directly. The phylogenetic analysis results showed that all of the 16 isolated strains were CRF01_AE recombinant form, and two clusters were set up in the phylogenetic tree. The tropic prediction of 16 strains showed that 2 isolates were CCR5 tropic, and the others are CXCR4 tropic. Eight of the isolated strains are drug resistant according to the genetic prediction. These 16 near full-length characterized CRF01_AE isolates obtained in this study will provide valuable genomic and phenotypic information on HIV-1 strains circulating in China for related researches.

Pseudolysobacter antarcticus gen. nov., sp. nov., isolated from soil in Fildes Peninsula, Antarctica.

A novel strain, designated AQ6-296T, was isolated from a soil sample collected in Fildes Peninsula, Antarctic. Cells were Gram-stain-negative, non-endospore-forming, non-motile, strictly aerobic and rod-shaped. Growth occurred at 4-28 °C (optimum, 20 °C) and at pH 6.0-7.0 (optimum, pH 7.0). NaCl was not obligatory for growth. Colonies were pale yellow after growth for 3 days at 20 °C on Reasoner's 2A agar. The strain was weakly positive for oxidase and the catalase test was negative. The only respiratory quinone was Q-8. The predominant cellular fatty acids were iso-C16 : 0, iso-C15 : 0, iso-C11 : 0 3OH, summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and summed feature 9 (comprising iso-C17 : 1ω9c and/or C16 : 010-methyl). The major polar lipids were phosphatidylethanolamine, unknown aminolipids, phosphatidylglycerol and diphosphatidylglycerol. The results of phylogenetic analysis based on 16S rRNA gene sequences (the highest similarity at 92.4 % to Lysobacter dokdonensis) indicated that strain AQ6-296T is within the family Xanthomonadaceae. The DNA G+C content of the type strain was 58.6 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain AQ6-296T is considered to represent a novel genus and species in the family Xanthomonadaceae, for which the name Pseudolysobacter antarcticus gen. nov., sp. nov. is proposed. The type strain is AQ6-296T (CCTCC AB 2016313T=KCTC 52744T).

Deinococcus psychrotolerans sp. nov., isolated from soil on the South Shetland Islands, Antarctica.

A Gram-stain-negative, non-motile, strictly aerobic, coccus-shaped bacterium, designated S14-83T, was isolated from a soil sample collected from the South Shetland Islands of Antarctica. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain is a novel member of the genus Deinococcus, with Deinococcus alpinitundrae as its closest relative (96.1 % similarity). The DNA G+C content of the strain was 61.1 mol% and the major respiratory quinone was MK-8. Major cellular fatty acids were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c) and C16 : 0. As well as containing glycophospholipid, aminophospholipids and glycolipid as major polar lipids, there were also some unknown polar lipids. The diagnostic diamino acid in the cell-wall peptidoglycan was ornithine, corroborating the assignment of the strain to the genus Deinococcus. Strain S14-83T was shown to be extremely resistant to gamma radiation (>10 kGy) and UV light (460 Jm-2). On the basis of phylogenetic, chemotaxonomic and phenotypic data presented here, strain S14-83T represents a novel species of the genus Deinococcus, for which the name Deinococcus psychrotolerans sp. nov. is proposed. The type strain is S14-83T (=CCTCC AB 2015449T= DSM 105285 T).

Deinococcus taklimakanensis sp. nov., isolated from desert soil.

A gamma- and UV radiation-tolerant, Gram-negative, short-rod-shaped bacterial strain, designated X-121T, was isolated from soil samples collected from the Taklimakan desert in Xinjiang, China. Strain X-121T showed the highest 16S rRNA gene sequence similarity with Deinococcus depolymerans TDMA-24T (94.7 %). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain X-121T is a member of a novel species belonging to the clade formed by members of the genus Deinococcus in the family Deinococcaceae. The DNA G+C content of strain X-121T was 63.6 mol%. The chemotaxonomic charateristics of strain X-121T were typical of members of the genus Deinococcus, with MK-8 being the predominant respiratory quinone, summed feature 3 (16 : 1ω7c,16 : 1ω6c), 16 : 0 and 17 : 1ω8c as major cellular fatty acid, several unidentified phosphoglycolipids and glycolipids as the dominant polar lipids, galactose as the predominant cell-wall sugar and the presence of peptidoglycan with l-ornithine. Strain X-121T is therefore identified as representing a novel species, for which the name Deinococcus taklimakanensis sp. nov. is proposed, with the type strain X-121T(=CCTCC AB 207228T=KCTC 33842T).

Sphingomonas antarctica sp. nov., isolated from Antarctic tundra soil.

Strain 200T, isolated from a soil sample taken from Antarctic tundra soil around Zhongshan Station, was found to be a Gram-stain-negative, yellow-pigmented, catalase-positive, oxidase-negative, non-motile, non-spore-forming, rod-shaped and aerobic bacterium. Strain 200T grew optimally at pH 7.0 and in the absence of NaCl on R2A. Its optimum growth temperature was 20 °C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 200T belonged to the genus Sphingomonas. Strain 200T showed the highest sequence similarities to Sphingomonas kyeonggiense THG-DT81T (95.1 %) and Sphingomonas molluscorum KMM 3882T (95.1 %). Chemotaxonomic analysis showed that strain 200T had characteristics typical of members of the genus Sphingomonas. Ubiquinone 10 was the predominant respiratory quinone and sym-homospermidine was the polyamine. The major polar lipids were sphingoglycolipid, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. The G+C content of the genomic DNA was determined to be 60.9 mol%. Strain 200T contained C16 : 0 (31.6 %), summed feature 8 (comprising C18 : 1ω7c and/or C18 : 1ω6c, 22.7 %), summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c, 11.2 %), C18 : 0 (7.8 %) and C14 : 0 2OH (6.7 %) as the major cellular fatty acids. On the basis of phylogenetic analysis, and physiological and biochemical characterization, strain 200T should be classified as representing a novel species of the genus Sphingomonas, for which the name Sphingomonasantarctica sp. nov. is proposed. The type strain is 200T (=CCTCC AB 2016064T=KCTC 52488T).

Flavitalea antarctica sp. nov., isolated from Fildes Peninsula, Antarctica.

A bright-yellow, Gram-stain-negative, rod-shaped, gliding and aerobic bacterium, designated strain AQ6-291T, was isolated from the Fildes Peninsula, Antarctica, and its taxonomic position was investigated by genotypic, phenotypic and chemotaxonomic analyses. Growth occurred at 4-28 °C (optimum 20 °C) and at pH 5.0-8.0 (optimum pH 7.0). Strain AQ6-291T contained iso-C15 : 1 G, iso-C15 : 0, C16 : 1ω5c, iso-C17 : 0 3-OH and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) as the major cellular fatty acids. The main polar lipids were phosphatidylethanolamine, unknown aminophospholipids, unknown phospholipids, five unknown aminolipids and two unknown polar lipids. MK-7 was the major respiratory quinone. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain AQ6-291T belonged to the genus Flavitalea. The DNA G+C content was 48.1 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain AQ6-291T is considered to represent a novel species of the genus Flavitalea, for which the name Flavitalea antarctica sp. nov. is proposed. The type strain is AQ6-291T (=CCTCC AB 2016109T=KCTC 52491T).

Cdk1 activity is required for mitotic activation of aurora A during G2/M transition of human cells.

In mammalian cells entry into and progression through mitosis are regulated by multiple mitotic kinases. How mitotic kinases interact with each other and coordinately regulate mitosis remains to be fully understood. Here we employed a chemical biology approach using selective small molecule kinase inhibitors to dissect the relationship between Cdk1 and Aurora A kinases during G(2)/M transition. We find that activation of Aurora A first occurs at centrosomes at late G(2) and is required for centrosome separation independently of Cdk1 activity. Upon entry into mitosis, Aurora A then becomes fully activated downstream of Cdk1 activation. Inactivation of Aurora A or Plk1 individually during a synchronized cell cycle shows no significant effect on Cdk1 activation and entry into mitosis. However, simultaneous inactivation of both Aurora A and Plk1 markedly delays Cdk1 activation and entry into mitosis, suggesting that Aurora A and Plk1 have redundant functions in the feedback activation of Cdk1. Together, our data suggest that Cdk1, Aurora A, and Plk1 mitotic kinases participate in a feedback activation loop and that activation of Cdk1 initiates the feedback loop activity, leading to rapid and timely entry into mitosis in human cells. In addition, live cell imaging reveals that the nuclear cycle of cells becomes uncoupled from cytokinesis upon inactivation of both Aurora A and Aurora B kinases and continues to oscillate in a Cdk1-dependent manner in the absence of cytokinesis, resulting in multinucleated, polyploidy cells.

Discovery of cercosporamide, a known antifungal natural product, as a selective Pkc1 kinase inhibitor through high-throughput screening.

The Pkc1-mediated cell wall integrity-signaling pathway is highly conserved in fungi and is essential for fungal growth. We thus explored the potential of targeting the Pkc1 protein kinase for developing broad-spectrum fungicidal antifungal drugs through a Candida albicans Pkc1-based high-throughput screening. We discovered that cercosporamide, a broad-spectrum natural antifungal compound, but previously with an unknown mode of action, is actually a selective and highly potent fungal Pkc1 kinase inhibitor. This finding provides a molecular explanation for previous observations in which Saccharomyces cerevisiae cell wall mutants were found to be highly sensitive to cercosporamide. Indeed, S. cerevisiae mutant cells with reduced Pkc1 kinase activity become hypersensitive to cercosporamide, and this sensitivity can be suppressed under high-osmotic growth conditions. Together, the results demonstrate that cercosporamide acts selectively on Pkc1 kinase and, thus, they provide a molecular mechanism for its antifungal activity. Furthermore, cercosporamide and a beta-1,3-glucan synthase inhibitor echinocandin analog, by targeting two different key components of the cell wall biosynthesis pathway, are highly synergistic in their antifungal activities. The synergistic antifungal activity between Pkc1 kinase and beta-1,3-glucan synthase inhibitors points to a potential highly effective combination therapy to treat fungal infections.

Synthesis of 1,7-annulated indoles and their applications in the studies of cyclin dependent kinase inhibitors.

The synthesis of a novel series of 1,7-annulated indolocarbazoles 2 and 16 is described. These compounds were found to be potent cyclin dependent kinase inhibitors with good antiproliferative activity against two human carcinoma cell lines. These inhibitors also arrested tumor cells at the G1 phase and inhibited pRb phosphorylation.

Aryl[a]pyrrolo[3,4-c]carbazoles as selective cyclin D1-CDK4 inhibitors.

The synthesis of new analogues of Arcyriaflavin A in which one indole ring is replaced by an aryl or heteroaryl ring is described. These new series of aryl[a]pyrrolo[3,4-c]carbazoles were evaluated as inhibitors of Cyclin D1-CDK4. A potent and selective D1-CDK4 inhibitor, 7a (D1-CDK4 IC(50)=45 nM), has been identified. The potency, selectivity profile against other kinases, and structure-activity relationship (SAR) trends of this class of compounds are discussed.

Studies on cyclin-dependent kinase inhibitors: indolo-[2,3-a]pyrrolo[3,4-c]carbazoles versus bis-indolylmaleimides.

A series of indolo[2,3-a]pyrrolo[3,4-c]carbazoles and their bis-indolylmaleimides precursors have been prepared in order to compare their activity as D1-CDK4 inhibitors. Both enzymatic and antiproliferative assays have shown that the structurally more constrained indolo[2,3-a]pyrrolo[3,4-c]carbazoles are consistently more active (8-42-fold) in head-to-head comparison with their bis-indolylmaleimides counterparts. Cell-cycle analysis using flow cytometry have also shown that the indolocarbazoles are selective G1 blockers while the bis-indolylmaleimides arrest cells in the G2/M phase.

Novel, potent and selective cyclin D1/CDK4 inhibitors: indolo[6,7-a]pyrrolo[3,4-c]carbazoles.

The synthesis and CDK inhibitory properties of a series of indolo[6,7-a]pyrrolo[3,4-c]carbazoles is reported. In addition to their potent CDK activity, the compounds display antiproliferative activity against two human cancer cell lines. These inhibitors also effect strong G1 arrest in these cell lines and inhibit Rb phosphorylation at Ser780 consistent with inhibition of cyclin D1/CDK4.

Synthesis, structure-activity relationship, and biological studies of indolocarbazoles as potent cyclin D1-CDK4 inhibitors.

Novel substituted indolocarbazoles were synthesized, and their kinase inhibitory capability was evaluated in vitro. 6-Substituted indolocarbazoles 4 were found to be potent and selective D1/CDK4 inhibitors. 4d and 4h exhibited potent and ATP-competitive D1/CDK4 activities with IC50 values of 76 and 42 nM, respectively. Both compounds had high selectivity against the other kinases. These D1/CDK4 inhibitors inhibited tumor cell growth, arrested tumor cells at the G1 phase, and inhibited pRb phosphorylation.

Synthesis of quinolinyl/isoquinolinyl[a]pyrrolo [3,4-c] carbazoles as cyclin D1/CDK4 inhibitors.

A novel series of pyrrolo[3,4-c] carbazoles fused with a quinolinyl/isoquinolinyl moiety were synthesized and their D1/CDK4 inhibitory and antiproliferative activity were evaluated. Compound 8H, 14H-isoquinolinyl[6,5-a]-pyrrolo[3,4-c]carbazole-7,9-dione (1d) was found to be a highly potent D1/CDK4 inhibitor with an IC(50) of 69 nM. Compound 1d also inhibited tumor cell growth, arrested tumor cells in G1 phase and inhibited pRb phosphorylation.