Journal: Cell Host & Microbe

Article Title: Complex Immune Dysregulation in COVID-19 Patients with Severe Respiratory Failure

doi: 10.1016/j.chom.2020.04.009

Figure Lengend Snippet:

Article Snippet: Human IFN-γ ELISA , Diaclone , 950.000.192.

Techniques: Control, Recombinant, Saline, Enzyme-linked Immunosorbent Assay, Software

Journal: Proceedings of the Japan Academy. Series B, Physical and Biological Sciences

Article Title: Purple pigments, catechinopyranocyanidins A and B, in the seed coats of red adzuki beans elute during processing to yield purple-colored an-paste

doi: 10.2183/pjab.101.037

Figure Lengend Snippet: HPLC chromatograms of extracts of dried adzuki bean (cv. Shumari) and lyophilized sarashi-an obtained by beans cv. Shumari. Black lines: absorbance at 280 nm; purple lines: absorbance at 570 nm. * indicates purple pigment of different structures from 1 and 2 . (A) Dried adzuki bean extract. (B) Lyophilized sarashi-an with general processing. (C) Lyophilized sarashi-an with pre-vacuum processing.

Article Snippet: Analytical HPLC was performed on a JASCO HPLC system (Jasco, Hachioji, Japan) comprising two pumps, along with a mixer, a degasser, a PDA detector, a column oven, and an autosampler, all controlled using ChromNAV 2.0 HPLC software (Jasco, Hachioji, Japan) on a personal computer, as reported previously. )

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Characterization of SPP inhibitors suppressing propagation of HCV and protozoa

doi: 10.1073/pnas.1712484114

Figure Lengend Snippet: YO-01027 is a potent inhibitor of the production of infectious particles of HCV. (A) Huh7 cells infected with HCV JFH1 strain at an MOI of 5 were treated with LY-411575 or YO-01027 for 4 d. Infectious titers in the culture supernatants were determined by a focus-forming assay. The P values were determined by a Student's t test. The IC99 values were calculated by using GraphPad Prism software. FFU, focus-forming units. (B) Huh7 cells infected with HCV JFH1 strains at an MOI of 5 were treated with the SPP inhibitors LY-411575 (0, 300, 1,000, or 3,000 nM) or YO-01027 (0, 30, 100, or 300 nM) and the DAAs telaprevir (TPV) or daclatasvir (DCV) for 4 d. Infectious titers in the culture supernatants were determined by a focus-forming assay. (C) Fractional IC99 (FC) values were calculated as the IC99 of an inhibitor combination divided by the IC99 of an inhibitor alone. The effect of synergy was investigated by isobologram analysis. (D) SPPKOHuh7 cells were lentivirally transduced with hsSPP and pfSPP. SPPKOHuh7 cells stably expressing either hsSPP (Hs) or pfSPP (Pf) were selected with puromycin, and the expression of hsSPP and pfSPP was confirmed by Western blotting. (E) Huh7 and SPPKOHuh7 cells stably expressing hsSPP (Hs) and pfSPP (Pf) were infected with a lentivirus expressing FLAG-tagged HCV core protein. The mature core protein was detected by Western blotting. (F) SPPKOHuh7 cells stably expressing hsSPP (Hs) and pfSPP (Pf) were infected with HCV at an MOI of 5. Infectious titers in the culture supernatants were determined by a focus-forming assay at 4 d postinfection. The P values were determined by a Student's t test. (G) SPPKOHuh7 cells stably expressing pfSPP were infected with lentivirus vector expressing HCV core protein and were treated with YO-01027 at a concentration of 0.1, 0.3, 1.0, or 3.0 µM. The matured core protein was detected by immunoblotting. (H) Treatment with YO-01027 at a concentration of 1, 10, or 20 μM for 4 d suppressed the production of infectious particles of HCV in SPPKOHuh7 cells expressing pfSPP upon infection with HCV at an MOI of 5. The P values were determined by a Student's t test. (I) The growth rate of P. falciparum treated with YO-01027 or LY-411575 was measured. The P values were determined by a Student's t test. (J) MEFs were infected with the T. gondii strain ME-49, which possesses a luciferase gene, at an MOI of 1 and then were incubated with the inhibitors for 1 d. The luminescent signal was measured. The P values were determined by a Student's t test. (K) MEFs, SPP-KO MEFs (SPPKOMEFs), Huh7 cells, and SPPKOHuh7 cells were infected with T. gondii at an MOI of 1. The luminescent signal was measured at 1 d postinfection. The P values were determined by a Student's t test. ns, not significant. (L) BALB/c mice i.p. infected with T. gondii (RH strain: 1 × 102 parasites) were i.p. treated with 5 μmol/kg or 10 μmol/kg of YO-01027 (n = 6 mice for each treatment) or vehicle (n = 6 mice) at 24 h postinfection. Bioluminescent analysis was performed at the indicated time points to visualize parasite growth. (M) The survival of mice was monitored. The P values were determined by a log-rank text. Data are the mean ± SD of two independent experiments performed with cell lines (A, B, F, and H–K) and are representative of three (G) or two (D, E, L, and M) independent experiments. Significant differences are indicated by double asterisks (**P < 0.01) or a single asterisk (*P < 0.05).

Article Snippet: RO-4929097, avagacestat, LY-450139, DAPT, MK-0752, telaprevir (VX-950), and daclatasvir (BMS-790052) were purchased from Selleck Chemicals.

Techniques: Infection, Focus Forming Assay, Software, Transduction, Stable Transfection, Expressing, Western Blot, Plasmid Preparation, Concentration Assay, Luciferase, Incubation

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Characterization of SPP inhibitors suppressing propagation of HCV and protozoa

doi: 10.1073/pnas.1712484114

Figure Lengend Snippet: SPP inhibitors are broad-spectrum antivirals against HCV of all genotypes with a low risk of emergence of drug-resistant breakthrough viruses. (A) Inhibitory effects of YO-01027 against the processing of core proteins of various genotypes of HCV. FLAG-tagged core proteins of GT1 (TN strain), GT1 (H77C strain), GT2 (J6 strain), GT2 (J8 strain), GT2 (JFH1 strain), GT3 (S52 strain), GT4 (ED43 strain), GT5 (SA13 strain), GT6 (HK6a strain), and GT7 (QC69 strain) were expressed in Huh7 cells and were treated with YO-01027 at a concentration of 0.1, 0.3, 1.0, or 3.0 µM. HCV core protein was detected by immunoblotting. Images are representative of at least three independent experiments. (B) Culture supernatant of Huh7.5.1 cells infected with HCV in the presence of LY-411575 (1.0 µM), YO-01027 (0.1 µM), or telaprevir (TPV, 0.1 µM) was passaged in naive cells in the presence of these reagents. After five or 15 passages, the supernatants were collected, and the inhibitory effects of the reagents were determined. (C) After five passages in the presence or absence of telaprevir, culture supernatants were inoculated into Huh7.5.1 cells in the presence of various concentrations of telaprevir, and the infectious titers were determined 4 d postinfection. (D) After 15 passages in the presence or absence of LY-411575 or YO-01027, the culture supernatants were inoculated into Huh7.5.1 cells in the presence of various concentrations of the inhibitors, and infectious titers were determined 4 d postinfection. Data in C and D are the mean ± SD of two independent experiments. **P < 0.01. P values were determined by a Student's t test.

Article Snippet: RO-4929097, avagacestat, LY-450139, DAPT, MK-0752, telaprevir (VX-950), and daclatasvir (BMS-790052) were purchased from Selleck Chemicals.

Techniques: Concentration Assay, Western Blot, Infection