Journal: Reaction Chemistry & Engineering

Article Title: Data-rich process development of immobilized biocatalysts in flow

doi: 10.1039/d1re00298h

Figure Lengend Snippet: Fig. 5 Initial resin screen for galactose oxidase using 0.5 mL columns with the following resins: (A) GE Ni Sepharose 6FF (B) Ni-NTA Superflow (C) Nuvia IMAC – Ni (D) Nuvia IMAC – Cu. Gradient profile is shown by dashed lines.

Article Snippet: Normalized relative area and retention are normalized relative to Bio-rad Nuvia IMAC Ni Resin Metal Base bead Normalized relative area % Rs Specificity Normalized retention GE Sepharose 6FF Ni Agarose 42% 2.53 24 0.993 Qiagen Ni-NTA Superflow Ni Agarose 51% 2.01 24 0.992 Bio-rad Nuvia IMAC Ni UNOsphere 100% 1.92 45 1.00 Bio-rad Nuvia IMAC Cu UNOsphere 143% 2.44 18 0.987 O pe n A cc es s A rt ic le .

Techniques:

Journal: Nature Communications

Article Title: Ciclopirox inhibits Hepatitis B Virus secretion by blocking capsid assembly

doi: 10.1038/s41467-019-10200-5

Figure Lengend Snippet: Effect of FDA-approved compounds on HBV replication. a Schematic depiction of the primary screening analysis. HepG2.2.15 cells, which were stably transfected with HBV replicons, were treated with 978 FDA-approved drugs (1 μM) for 3 days. Entecavir (ETV) (1 μM) served as positive control. The effects of the compounds on secretion of HBV DNA particles were assessed by quantitative PCR with an HBV DNA primer and probe set. b The 19 candidates that responded well in a were tested again in HepG2.2.15 cells for 6 days instead of 3 days. c The 19 candidates were assessed for their ability to inhibit HBV pgRNA and precoreRNA transcription in HepG2.2.15 cells. Quantitative RT-PCR was performed after 6 days of incubation. d Effect of the 19 candidates on intracellular HBV capsid assembly. Huh-7 cells were transfected with pCDNA3-Core and exposed after 12 h to the 19 candidates (1 μM) for 36 h. The cells were lysed and assembled HBV core particles were isolated by sucrose step gradient ultracentrifugation, resolved on 1% agarose gels, and subjected to immunoblot analysis with anti-HBV core antibody. The data shown in b – d are representative of three independent experiments and are expressed as means ± SDs. The error bars represent the ± SD. * p < 0.05; ** p < 0.01, as determined by unpaired two-tailed Student’s t -tests. Source data are provided as a Source Data file

Article Snippet: Liver tissue samples were analyzed to measure cccDNA, and sectioned for immunohistochemical staining with anti-HBV core antibodies (OAAI00128, Aviva Systems Biology).

Techniques: Stable Transfection, Transfection, Positive Control, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Incubation, Isolation, Western Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Ciclopirox inhibits Hepatitis B Virus secretion by blocking capsid assembly

doi: 10.1038/s41467-019-10200-5

Figure Lengend Snippet: Ciclopirox inhibits in vitro and intracellular HBV capsid assembly. a Effects of ciclopirox, GLS-4, and Bay41–4109 on in vitro HBV capsid assembly. Various concentrations of ciclopirox, GLS-4, or Bay41–4109 were mixed with Cp149 in reaction buffer. After 1 h, the mixtures were subjected to a immunoblot analysis with anti-HBV core antibody. b Sucrose density gradient analysis of capsid assembly inhibition by ciclopirox. Ten fractions collected from top to bottom underwent immunoblot analysis. c , d Effect of ciclopirox on intracellular HBV capsid assembly. Huh-7 cells were transfected with ( c ) pCDNA3-Core or ( d ) pHBV1.2× and exposed after 12 h to the indicated concentrations of ciclopirox for 36 h. The cells were lysed and assembled HBV core particles were isolated by sucrose step gradient ultracentrifugation, resolved on 1% agarose gels, and subjected to immunoblot analysis with anti-HBV core antibody. e Electron micrographs of the effect of ciclopirox on in vitro HBV capsid assembly. The particles were negatively stained with 2% uranyl acetate. Scale bars = 200 nm and 100 nm. TEM magnifications, ×49,000 and ×75,000. Ciclopirox appeared to convert the T = 4 HBV capsid into T = 3 form (30 nm). The data shown in a – e are representative of three independent experiments and are expressed as mean ± SD. The error bars represent the ± SD. * p < 0.05; ** p < 0.01, as determined by unpaired two-tailed Student’s t -tests. Source data are provided as a Source Data file

Article Snippet: Liver tissue samples were analyzed to measure cccDNA, and sectioned for immunohistochemical staining with anti-HBV core antibodies (OAAI00128, Aviva Systems Biology).

Techniques: In Vitro, Western Blot, Inhibition, Transfection, Isolation, Staining, Two Tailed Test

Journal: Nature Communications

Article Title: Ciclopirox inhibits Hepatitis B Virus secretion by blocking capsid assembly

doi: 10.1038/s41467-019-10200-5

Figure Lengend Snippet: Ciclopirox inhibits HBV replication in an in vitro HBV infection model. a Schematic depiction of the experiment. HepG2-NTCP and Huh-7-NTCP cells were treated with ciclopirox for 6 h and exposed to HBV for 16 h. After washing, the cells were treated daily with ciclopirox and cultured for another 14 days. b NTCP protein expression analyzed by immunoblot analysis. c GFP protein expression analyzed by flow cytometry. d – f Quantitative PCR with an HBV DNA primer and probe set was used to measure secreted HBV DNA ( d ), intracellular cccDNA ( e ), and intracellular HBV rcDNA ( f ). Intracellular cccDNA was also detected by southern blot analysis ( e ). The data in b – f are representative of three independent experiments and are expressed as mean ± SD. The error bars represent the ± SD. * p < 0.05; ** p < 0.01, by unpaired two-tailed Student’s t -tests. Source data are provided as a Source Data file

Article Snippet: Liver tissue samples were analyzed to measure cccDNA, and sectioned for immunohistochemical staining with anti-HBV core antibodies (OAAI00128, Aviva Systems Biology).

Techniques: In Vitro, Infection, Cell Culture, Expressing, Western Blot, Flow Cytometry, Real-time Polymerase Chain Reaction, Southern Blot, Two Tailed Test

Journal: Nature Communications

Article Title: Ciclopirox inhibits Hepatitis B Virus secretion by blocking capsid assembly

doi: 10.1038/s41467-019-10200-5

Figure Lengend Snippet: Antiviral activity of ciclopirox in a humanized liver mouse model of HBV infection. a Schematic depiction of the experiment. PXB mice ( n = 3 per group) were injected via the tail vein with HBV virions (5 × 10 7 copies per mouse, genotype D). After 6 weeks, the mice were treated orally with TDF and/or ciclopirox (both 5 mg per kg, daily), and serum samples were taken from the orbital sinus every week. b Serum HBV genomic DNA levels measured by quantitative PCR. c Serum HBsAg levels determined by ELISA. d Serum HBeAg levels determined by ELISA. e Serum ALT levels determined by ELISA. f Human albumin levels determined by ELISA. g Intrahepatic cccDNA was determined in nucleic acid extracted from liver specimens obtained after 35 days of treatment. h Immunohistochemical analysis of hepatitis B core antigen (HBcAg) in liver sections obtained after 35 days of treatment (×200 magnification). The data in b – g are representative of three independent experiments, and are expressed as means ± SDs. The error bars represent the ± SD. Scale bars, 100 μm.* p < 0.05; ** p < 0.01, by unpaired two-tailed Student’s t -tests. Source data are provided as a Source Data file

Article Snippet: Liver tissue samples were analyzed to measure cccDNA, and sectioned for immunohistochemical staining with anti-HBV core antibodies (OAAI00128, Aviva Systems Biology).

Techniques: Activity Assay, Infection, Injection, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Two Tailed Test

Journal: Canadian Journal of Gastroenterology & Hepatology

Article Title: Applicability of Oral Fluid and Dried Blood Spot for Hepatitis B Virus Diagnosis

doi: 10.1155/2019/5672795

Figure Lengend Snippet: Main Characteristics of studies that detect HBV antigen and antibodies in saliva or oral fluid samples.

Article Snippet: Anti-HBc Total , Viral Hepatitis ambulatory , Brazil , Orasure , Commercial Enzyme immunoassay , 13% , 100.0% , [ ] .

Techniques: Enzyme-linked Immunosorbent Assay