Journal: Acta Pharmaceutica Sinica. B

Article Title: Research and development of Chinese anti-COVID-19 drugs

doi: 10.1016/j.apsb.2022.09.002

Figure Lengend Snippet: The anti-COVID-19 drugs approved for clinical research in China.

Article Snippet: , SCTA01 (HB27) , Sinocelltech Ltd. , RBD , Intravenous injection , Severe , Phase Ⅱ/Ⅲ clinical study , Antiviral effect.

Techniques: Injection, Recombinant, Derivative Assay, Capsules, Plasmid Preparation

Journal: Acta Pharmaceutica Sinica. B

Article Title: Research and development of Chinese anti-COVID-19 drugs

doi: 10.1016/j.apsb.2022.09.002

Figure Lengend Snippet: The main anti-COVID-19 drugs under research and development by China.

Article Snippet: , SCTA01 (HB27) , Sinocelltech Ltd. , RBD , Intravenous injection , Severe , Phase Ⅱ/Ⅲ clinical study , Antiviral effect.

Techniques: Control, Injection, Capsules, Infection

Journal: Journal of Bacteriology

Article Title: Mlc of Thermus thermophilus : a Glucose-Specific Regulator for a Glucose/Mannose ABC Transporter in the Absence of the Phosphotransferase System

doi: 10.1128/jb.00715-06

Figure Lengend Snippet: FIG. 4. Mlc glucose/mannose transport operon and TMSP ABC transport operon organization in T. thermophilus. The TTC numbers refer to the gene numbering in the genome sequencing of T. thermophilus HB27 (10). We propose the names GlcE, GlcF, and GlcG for the glucose/ mannose-binding protein and the two membrane components of the glucose/mannose ABC transporter.

Article Snippet: Strains and plasmids used in this study Strain or plasmid Known genotypea Source orreference Strains E. coli SF120 ptr32::cm degP4::kan ompT 2 JM-G2 ptsG-lacZ mlc::Tn10 22 T. thermophilus DSM7029 Wild-type strain (HB27) DSMZ CL3 mlc::kan This study HB27 malK1::kan malK1::kan 28 JN1 malF::bleo Jutta Nesper CL4 mlc::kan malF::bleo This study Plasmids pFC4 pGDR11 lacIq; N-terminally His6-tagged MlcTth (gene TTC0329); Amp r This study pEM1 pCS19 lacIq; wild-type MlcTth (gene TTC0329); Amp r This study pCL2 pQE30 (QIAGEN) lacIq; N-terminally His6-tagged Mlc916Tth::Kan; Amp r Kanr This study pTTC0328 pQE30 (QIAGEN) lacIq; N-terminally His6-tagged glucose-binding protein (gene TTC0328) without its signal sequence This study pCL10 pQE30(QIAGEN) lacIq; N-terminal His6-tagged TMSP transporter binding protein (gene TTC1627) without its signal sequence This study pCS19 pQE60 (QIAGEN) lacIq; Ampr 29 pGDR11 pQE31 (QIAGEN) lacIq; N-terminal His6 tag; Amp r 20 pMK18 pUC18 derivative with a thermostable resistance to kanamycin 6 pREP4 pACYC derivative containing the p15A replicon; Kanr QIAGEN on M arch 7, 2015 by M A H ID O L U N IV F A C O F M E D http://jb.asm .org/ D ow nloaded from ampicillin/ml and 25 g of kanamycin/ml.

Techniques: Sequencing, Binding Assay, Membrane

Journal: Journal of Bacteriology

Article Title: Mlc of Thermus thermophilus : a Glucose-Specific Regulator for a Glucose/Mannose ABC Transporter in the Absence of the Phosphotransferase System

doi: 10.1128/jb.00715-06

Figure Lengend Snippet: FIG. 7. Glucose transport kinetics of T. thermophilus. (A) Wild type (HB27); (B) malF::bleo mutant (JN1); (C) mlc::kan mutant (CL3). Transport by JN1 represents the activity of the glucose/man- nose ABC transporter, and transport by CL3 represents the activity of the TMSP ABC transporter. The curves can be fitted according to the Michaelis-Menten equation and yield a Km of 0.15 M for glucose uptake via the glucose/mannose transporter and a Km of 1.4 M for the TMSP transporter. A double mutant harboring both malF::bleo and mlc::kan exhibits neither glucose nor maltose transport activity. TABLE 2. Transport of glucose and maltose in T. thermophilus

Article Snippet: Strains and plasmids used in this study Strain or plasmid Known genotypea Source orreference Strains E. coli SF120 ptr32::cm degP4::kan ompT 2 JM-G2 ptsG-lacZ mlc::Tn10 22 T. thermophilus DSM7029 Wild-type strain (HB27) DSMZ CL3 mlc::kan This study HB27 malK1::kan malK1::kan 28 JN1 malF::bleo Jutta Nesper CL4 mlc::kan malF::bleo This study Plasmids pFC4 pGDR11 lacIq; N-terminally His6-tagged MlcTth (gene TTC0329); Amp r This study pEM1 pCS19 lacIq; wild-type MlcTth (gene TTC0329); Amp r This study pCL2 pQE30 (QIAGEN) lacIq; N-terminally His6-tagged Mlc916Tth::Kan; Amp r Kanr This study pTTC0328 pQE30 (QIAGEN) lacIq; N-terminally His6-tagged glucose-binding protein (gene TTC0328) without its signal sequence This study pCL10 pQE30(QIAGEN) lacIq; N-terminal His6-tagged TMSP transporter binding protein (gene TTC1627) without its signal sequence This study pCS19 pQE60 (QIAGEN) lacIq; Ampr 29 pGDR11 pQE31 (QIAGEN) lacIq; N-terminal His6 tag; Amp r 20 pMK18 pUC18 derivative with a thermostable resistance to kanamycin 6 pREP4 pACYC derivative containing the p15A replicon; Kanr QIAGEN on M arch 7, 2015 by M A H ID O L U N IV F A C O F M E D http://jb.asm .org/ D ow nloaded from ampicillin/ml and 25 g of kanamycin/ml.

Techniques: Mutagenesis, Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: The 2′- O -methylation status of a single guanosine controls transfer RNA–mediated Toll-like receptor 7 activation or inhibition

doi: 10.1084/jem.20111075

Figure Lengend Snippet: tRNA from different bacterial species varies in their IFN-α–inducing potential. (A) Purified tRNA from different bacterial species were visualized on a 12% polyacrylamide gel (lane 1: B. subtilis ; lane 2: P. aeruginosa ; lane 3: T. thermophilus ; lane 4: E. coli Nissle 1917 ; lane 5: L. lactis ; lane 6: M. marburgensis ). One representative gel is shown ( n > 3). (B) Human PBMCs were stimulated with 2 µg/ml of purified tRNA from different bacterial species as indicated and IFN-α production is depicted as percentage of activity of the TLR7 agonist RNA40. Experimental data were obtained from PBMCs preparations of 3–10 different donors per strain. Statistical analysis was performed using the paired Student’s t test (***, P < 0.0001; *, P = 0.02).

Article Snippet: The following strains were cultured in medium or agar plates as indicated: A. lwoffii F78 (provided by O. Holst and H. Heine, Leibniz-Zentrum Borstel, Borstel, Germany; 2× Lysogeny broth Lennox [LB]; Roth; 964.2×, 37°C, aerobic), B. subtilis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; Brain heart Infusion [BHI] medium; Oxoid Deutschland GmbH; CM0225, 37°C, aerobic), E. coli Nissle 1917 (isolated from Mutaflor; Ardeypharm; LB, 37°C, aerobic), H. influenzae b (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; chocolate blood; BD; 257011, 37°C, microaerophilic conditions), L. lactis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MRS medium; Oxoid Deutschland GmbH; CM0359/0361; 37°C, aerobic), M. marburgensis (provided by R. Thauer and J. Moll, MPI for Terrestrial Microbiology, Marburg, Germany; mineral medium; 80% H 2 /20% CO 2 /0.01% H 2 S, 65°C), P. aeruginosa (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MacConkey II-Agar; BD; 221172; 37°C, aerobic), S. aureus (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; LB medium; 37°C, aerobic), T. thermophilus HB27 (DSMZ; DSM7039; LB, 80°C, aerobic).

Techniques: Purification, Activity Assay

Journal: The Journal of Experimental Medicine

Article Title: The 2′- O -methylation status of a single guanosine controls transfer RNA–mediated Toll-like receptor 7 activation or inhibition

doi: 10.1084/jem.20111075

Figure Lengend Snippet: 2′- O -methylation at tRNA position G18 is responsible for non-immunostimulatory character of E. coli Nissle 1917 . (A) Human PBMCs were stimulated with 2, 0.4 and 0.1 µg/ml of purified tRNA from E. coli BW25113 – and E. coli BW25113 –derived knockout mutants for trmA (uracil-5-methyltransferase, m5U54), trmB (guanine-7-methyltransferase, m7G46), and trmH (Gm18-2′- O -methyltransferase). PBMCs were incubated with RNA for 20 h with subsequent IFN-α detection by ELISA. Representative experimental data from one out of four PBMC preparations are shown ( n = 2 ± SD). (B) HPLC analysis of P1 nuclease and phosphatase-treated tRNA from E. coli BW25113 , E. coli ΔtrmH, and E. coli ΔtrmH methylated in vitro with T. thermophilus trmH. (C) SDS-PAGE of purified recombinant T. thermophilus trmH protein expressed in E. coli BL21 . (D) Purified tRNA from E. coli ΔtrmH was incubated with 14 C-labeled SAM and with or without T. thermophilus trmH. tRNA was further visualized after PAGE by ethydium bromide (EtBr) staining and autoradiography (AR). For B–D, one representative experiment ( n ≥ 2) is shown. (E) Human PBMCs were stimulated with purified tRNA (2 µg/ml) from A. lwoffii , T. thermophilus , E. coli ΔtrmH, and E. coli ΔtrmH methylated in vitro with recombinant trmH and T. thermophilus . PBMCs were incubated with RNA for 20 h with subsequent IFN-α detection by ELISA. (F) Human enriched monocytes were stimulated with 100 ng/ml LPS, 1 µM CpG2216, 1 µg/ml RNA40, nuclease P1, and 2 µg/ml tRNA from T. thermophilus and A. lwoffii ± nuclease P1 digestion. After incubation for 20 h, IL-6 production was measured by ELISA. Representative experimental data from one out of four PBMCs (E) or four monocyte (F) preparations are shown ( n = 2 ± SD).

Article Snippet: The following strains were cultured in medium or agar plates as indicated: A. lwoffii F78 (provided by O. Holst and H. Heine, Leibniz-Zentrum Borstel, Borstel, Germany; 2× Lysogeny broth Lennox [LB]; Roth; 964.2×, 37°C, aerobic), B. subtilis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; Brain heart Infusion [BHI] medium; Oxoid Deutschland GmbH; CM0225, 37°C, aerobic), E. coli Nissle 1917 (isolated from Mutaflor; Ardeypharm; LB, 37°C, aerobic), H. influenzae b (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; chocolate blood; BD; 257011, 37°C, microaerophilic conditions), L. lactis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MRS medium; Oxoid Deutschland GmbH; CM0359/0361; 37°C, aerobic), M. marburgensis (provided by R. Thauer and J. Moll, MPI for Terrestrial Microbiology, Marburg, Germany; mineral medium; 80% H 2 /20% CO 2 /0.01% H 2 S, 65°C), P. aeruginosa (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MacConkey II-Agar; BD; 221172; 37°C, aerobic), S. aureus (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; LB medium; 37°C, aerobic), T. thermophilus HB27 (DSMZ; DSM7039; LB, 80°C, aerobic).

Techniques: Methylation, Purification, Derivative Assay, Knock-Out, Incubation, Enzyme-linked Immunosorbent Assay, In Vitro, SDS Page, Recombinant, Labeling, Staining, Autoradiography

Journal: The Journal of Experimental Medicine

Article Title: The 2′- O -methylation status of a single guanosine controls transfer RNA–mediated Toll-like receptor 7 activation or inhibition

doi: 10.1084/jem.20111075

Figure Lengend Snippet: Gm18-modified tRNA acts as antagonist for TLR7-mediated IFN-α production upon tRNA or ssRNA stimulation and viral infection. (A) Mouse FLT3L-induced DCs were stimulated with 2 µg/ml tRNA from A. lwoffii and various concentrations of tRNA from E. coli Nissle or T. thermophilus (2, 1.5, 1, 0.5, 0.2, 0.1, 0.05, 0.025, and 0.0125 µg/ml). (B) Human PBMCs were stimulated with 1 µg/ml RNA40 and various concentrations of tRNA from T. thermophilus or A. lwoffii (2, 0.5, 0.125, and 0.0313 µg/ml). (C) Human PBMCs were infected with various MOI (10, 1, and 0.1) of influenza virus A/PR/8/34 (IAV) and 2 µg/ml purified tRNA from T. thermophilus . For each stimulation, tRNA was complexed to DOTAP and incubated for 20 h with subsequent IFN-α detection by ELISA. For A–C, representative experimental data from one out of four responsive PBMCs or two mouse cell preparations are shown ( n = 2 ± SD).

Article Snippet: The following strains were cultured in medium or agar plates as indicated: A. lwoffii F78 (provided by O. Holst and H. Heine, Leibniz-Zentrum Borstel, Borstel, Germany; 2× Lysogeny broth Lennox [LB]; Roth; 964.2×, 37°C, aerobic), B. subtilis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; Brain heart Infusion [BHI] medium; Oxoid Deutschland GmbH; CM0225, 37°C, aerobic), E. coli Nissle 1917 (isolated from Mutaflor; Ardeypharm; LB, 37°C, aerobic), H. influenzae b (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; chocolate blood; BD; 257011, 37°C, microaerophilic conditions), L. lactis (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MRS medium; Oxoid Deutschland GmbH; CM0359/0361; 37°C, aerobic), M. marburgensis (provided by R. Thauer and J. Moll, MPI for Terrestrial Microbiology, Marburg, Germany; mineral medium; 80% H 2 /20% CO 2 /0.01% H 2 S, 65°C), P. aeruginosa (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; MacConkey II-Agar; BD; 221172; 37°C, aerobic), S. aureus (clinical isolate; Institute for Medical Microbiology and Hygiene in Marburg; LB medium; 37°C, aerobic), T. thermophilus HB27 (DSMZ; DSM7039; LB, 80°C, aerobic).

Techniques: Modification, Infection, Virus, Purification, Incubation, Enzyme-linked Immunosorbent Assay