Journal: Scientific reports

Article Title: A glycoside analog of mammalian oligomannose formulated with a TLR4-stimulating adjuvant elicits HIV-1 cross-reactive antibodies.

doi: 10.1038/s41598-021-84116-w

Figure Lengend Snippet: Figure 2. NIT211 conjugate is recognized by oligomannose-specific bnAbs and their germline precursors. NIT211 (4.1 glycosides per CRM197) was coated as solid-phase antigen onto ELISA plate wells at 5 µg/ml in PBS and assayed for recognition by the different antibodies. All antibodies were expressed recombinantly as human IgG1. (A) Binding of PGT128/130 bnAb family members PGT125, PGT126, PGT128 and PGT130. (B) Binding of non-PGT128/130 family antibodies BF520.1, BG18, PCDN-33A, PGDM12, PGDM21, PCDN76- 33A, VRC41.01. (C) Binding of inferred gl precursors BF520.1, BG18, PCDN-33A and the PGT128/130 family. Results are from single experiments performed with technical duplicates.

Article Snippet: Total serum IgG was detected with a mixture of equal amounts of alkaline phosphatase conjugated anti-mouse IgG1, IgG2b, IgG2c and IgG3 secondary antibodies (Jackson ImmunoResearch) and nitrophenylphosphate substrate (Sigma).

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Scientific reports

Article Title: A glycoside analog of mammalian oligomannose formulated with a TLR4-stimulating adjuvant elicits HIV-1 cross-reactive antibodies.

doi: 10.1038/s41598-021-84116-w

Figure Lengend Snippet: Figure 4. The relative binding affinities of PGT antibodies for NIT211 increase with increasing ligand density. Binding to NIT211 conjugates was assessed by ELISA. NIT211 (2.6 ligands, 4.1, and 6.2 ligands) was coated as solid-phase antigen onto ELISA plate wells at 5 µg/ml (82, 79, 76 nM respectively) and assayed for antibody binding. (A) Binding of PGT125, PGT126, PGT128 and PGT130 IgG to NIT211 at three different densities of glycoside per CRM197. (B) Binding of PGT125, PGT126, PGT128 and PGT130 Fabs to NIT211 at two different densities of glycoside per CRM197.

Article Snippet: Total serum IgG was detected with a mixture of equal amounts of alkaline phosphatase conjugated anti-mouse IgG1, IgG2b, IgG2c and IgG3 secondary antibodies (Jackson ImmunoResearch) and nitrophenylphosphate substrate (Sigma).

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: A glycoside analog of mammalian oligomannose formulated with a TLR4-stimulating adjuvant elicits HIV-1 cross-reactive antibodies.

doi: 10.1038/s41598-021-84116-w

Figure Lengend Snippet: Figure 5. Only animals administered GLA-SE-adjuvanted NIT211 mount an IgG response to the oligomannose mimetic that is of the IgG3 subclass. Trianni mice (n = 5 per group) were immunized subcutaneously (days 0, 21, 42 and 105) and sera collected on day 0 prior to immunization and on days 10, 28, 49, and 119 post- immunization. (A) Binding of total IgG antibodies in pre-immune and post-immune sera to BSA-conjugated oligomannose mimetic. Binding curves represent mean values for the five animals in each immunization group, each tested in duplicate, with error bars denoting the standard deviation from the mean. (B) Individual IgG binding curves for NIT211 + GLA-SE immunized mice (Ms1 to 5). The sex of each mouse is also noted. (C) IgG1, IgG2b, IgG2c and IgG3 antibody subclass responses in day 119 post-immune sera of NIT211 + GLA-SE immunized animals for the BSA-conjugated oligomannose mimetic in comparison to the CRM197 protein carrier.

Article Snippet: Total serum IgG was detected with a mixture of equal amounts of alkaline phosphatase conjugated anti-mouse IgG1, IgG2b, IgG2c and IgG3 secondary antibodies (Jackson ImmunoResearch) and nitrophenylphosphate substrate (Sigma).

Techniques: Binding Assay, Standard Deviation, Comparison

Journal: Scientific reports

Article Title: A glycoside analog of mammalian oligomannose formulated with a TLR4-stimulating adjuvant elicits HIV-1 cross-reactive antibodies.

doi: 10.1038/s41598-021-84116-w

Figure Lengend Snippet: Figure 6. NIT211 formulated in GLA-SE elicits antibodies with capacity to bind recombinant HIV Env trimers. (A) Binding of total IgG from day 119 sera from unimmunized, and NIT211-immunized animal groups to SOSIP trimers AMC008 (subtype B), Du422 (subtype C), ZM197M (subtype C), BG505 (subtype A), B41 (subtype B), and CZA97 (subtype C). The HIS-tagged trimers were captured on nickel-coated ELISA plates at 5 µg/ml in PBS. (B) Level of IgG1, IgG2, IgG3 antibody subclass binding (1:100 dilution) to B41 SOSIP trimer in day 119 sera of NIT211 + GLA-SE immunized animals. (C) Residual binding of bnAb PGT128 to B41 SOSIP trimer following incubation with sera from NIT211 + GLA-SE immunized animals (day 119) vs sera from KLH + Alum/CpG ODN1826 immunized animals (day 34). (D) Day 119 sera from NIT211 + GLA-SE immunized animals and sera from a group of unimmunized animals were assessed for pseudovirus neutralization using a panel of seven diverse HIV-1 strains (92TH021, 92RW020, 94UG103, 92BR020, 97ZA012, JRCSF and NL4-3). Pseudotyped vesicular stomatitis virus (VSV) was used as a negative control. All graphs depict mean values for the serum samples from all animals (n = 5) in each group. Error bars represent standard error from the mean.

Article Snippet: Total serum IgG was detected with a mixture of equal amounts of alkaline phosphatase conjugated anti-mouse IgG1, IgG2b, IgG2c and IgG3 secondary antibodies (Jackson ImmunoResearch) and nitrophenylphosphate substrate (Sigma).

Techniques: Recombinant, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation, Neutralization, Virus, Negative Control

Journal: Microbiology Spectrum

Article Title: Mechanism of Action of Isopropoxy Benzene Guanidine against Multidrug-Resistant Pathogens

doi: 10.1128/spectrum.03469-22

Figure Lengend Snippet: IBG restores the sensitivity of colistin against Gram-negative bacteria. (A) Chemical structure of the IBG. The guanidine group is marked in red. (B) Inhibition rate of colistin combined with IBG against 30 Gram-negative bacteria. (C and D) Time-kill assays conducted with colistin (0.5× MIC), IBG monotherapy (10 μg/mL), the combination, or later addition of colistin or IBG against colistin-susceptible E. coli ATCC 25922 (C) and colistin-resistant E. coli SHP45 (D).

Article Snippet: We found that colistin monotherapy produced highly resistant strains with a 64-fold increase in the MIC for ATCC 25922 and strains with a 4-fold increase in the MIC for SHP45.

Techniques: Bacteria, Inhibition

Journal: Microbiology Spectrum

Article Title: Mechanism of Action of Isopropoxy Benzene Guanidine against Multidrug-Resistant Pathogens

doi: 10.1128/spectrum.03469-22

Figure Lengend Snippet: Antibacterial activity of IBG ^a

Article Snippet: We found that colistin monotherapy produced highly resistant strains with a 64-fold increase in the MIC for ATCC 25922 and strains with a 4-fold increase in the MIC for SHP45.

Techniques: Activity Assay, Bacteria

Journal: Microbiology Spectrum

Article Title: Mechanism of Action of Isopropoxy Benzene Guanidine against Multidrug-Resistant Pathogens

doi: 10.1128/spectrum.03469-22

Figure Lengend Snippet: Evolution of colistin resistance in colistin-susceptible and colistin-resistant E. coli . The fold change of colistin MIC was detected after colistin-susceptible E. coli ATCC 25922 (A) and colistin-resistant E. coli SHP45 (B) were treated with 0.5× MIC colistin alone or in combination with IBG at 5 μg/mL or 10 μg/mL in vitro .

Article Snippet: We found that colistin monotherapy produced highly resistant strains with a 64-fold increase in the MIC for ATCC 25922 and strains with a 4-fold increase in the MIC for SHP45.

Techniques: In Vitro

Journal: Microbiology Spectrum

Article Title: Mechanism of Action of Isopropoxy Benzene Guanidine against Multidrug-Resistant Pathogens

doi: 10.1128/spectrum.03469-22

Figure Lengend Snippet: Antibacterial mechanism of IBG against MDR bacteria. (A) Dynamic curves of the outer membrane probed with NPN in E. coli ATCC 25922 treated with IBG (10 μg/mL), colistin (0.12 μg/mL), and both together. The fluorescence was detected with excitation and emission wavelengths of 350 nm and 420 nm. (B) The ΔpH after treatment with different concentrations of IBG (from 0 to 10× MIC) was determined in S. aureus ATCC 29213. The means for three biological replicates are shown, and error bars represent the SD. (C) Levels of intracellular ATP in S. aureus ATCC 29213 after treatment of IBG. Nonparametric one-way analysis of variance (ANOVA) was used to calculate P values (*, P < 0.05; **, P < 0.01). (D) Accumulation of ROS in E. coli ATCC 25922 treated with IBG with or without 0.12 μg/mL colistin. (E) IBG inhibits the transcript level of mcr -1 in E. coli SHP45 determined by qRT-PCR. All data are means and SD, and significance was determined by nonparametric one-way ANOVA (**, P < 0.01). (F) Scheme of mechanisms of action of IBG in Gram-positive and Gram-negative bacteria.

Article Snippet: We found that colistin monotherapy produced highly resistant strains with a 64-fold increase in the MIC for ATCC 25922 and strains with a 4-fold increase in the MIC for SHP45.

Techniques: Bacteria, Membrane, Fluorescence, Quantitative RT-PCR

Journal: Antimicrobial Agents and Chemotherapy

Article Title: In Vitro Antiviral Activity and Resistance Profile Characterization of the Hepatitis C Virus NS5A Inhibitor Ledipasvir

doi: 10.1128/AAC.02524-15

Figure Lengend Snippet: LDV is a specific inhibitor of HCV

Article Snippet: HeLa cells were obtained from the ATCC and infected with a mixture of three rhinovirus strains (human rhinovirus 1A [HRV1A], HRV16, and HRV14; ATCC, Manassas, VA).

Techniques: Virus

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 1. Western blots showing the positive and negative controls of NOXs 1-4 antibodies. Autoradiographs showing total NOX1 (A), NOX3 (C) and NOX4 (D) densities in cardiac ventricles from rats and normal hamsters (NH) as well as in A10 cells. In panel B, the autoradiograph shows the total NOX2 density at the cardiac ventricle level in NHs as well as in A10 cells. In panel D, HK cell extracts were used as positive controls for the NOX4 antibody. Negative controls were performed by incubating the membranes in the presence of the respective antibody blocked with the corresponding NOX peptide from the supplier. The band corresponding to the reference protein GAPDH shows the presence of proteins in the negative control wells.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot, Autoradiography, Negative Control

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 2. Decrease in NOX1 density during the development of heart failure in the hereditary cardiomyopathic hamster. (A) Autoradiographs showing NOX1 density in NH, HCMH and rat hearts. (B) Bar graph showing the decrease in NOX1/GAPDH ratio density in HCMH hearts compared to age-matched NH hearts as well as a lower NOX1/GAPDH ratio in rat hearts compared to NH hearts. The values are expressed as mean ± SEM where n is the number of Western blot wells of at least three different animals. *** P <0.001.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 3. Absence of change in NOX2 density during the development of heart failure in the hereditary cardiomyopathic hamster. (A) Autoradiographs showing NOX2 density in NH and HCMH hearts. (B) Bar graph showing that the NOX2/GAPDH ratio does not change in HCMH hearts compared to those from age- matched NHs. The values are expressed as mean ± SEM where n is the number of Western blot wells of at least three different animals.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 4. Increase in NOX3 density during the development of heart failure in the hereditary cardiomyopathic hamster. (A) Autoradiographs showing NOX3 density in NH, HCMH and rat hearts. (B) Bar graph showing the increase in NOX3/GAPDH ratio in HCMH hearts compared to those from age-matched NHs as well as a higher NOX3/GAPDH ratio in rat hearts compared to NH hearts. The values are expressed as mean ± SEM where n is the number of Western blot wells of at least three different animals.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 5. Decrease in NOX4 density during the development of heart failure in the hereditary cardiomyopathic hamster. (A) Autoradiographs showing NOX4 density in NH, HCMH and rat hearts. (B) Bar graph showing the decrease in NOX4/GAPDH ratio in HCMH hearts compared to age-matched NH hearts. However, there is no difference in NOX4/GAPDH ratio between rat and NH hearts. The values are expressed as mean ± SEM where n is the number of Western blot wells of at least three different animals. * P <0.05; ** P <0.01; *** P <0.001.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot

Journal: Canadian journal of physiology and pharmacology

Article Title: Increase of NADPH oxidase 3 in heart failure of hereditary cardiomyopathy 1 .

doi: 10.1139/cjpp-2019-0055

Figure Lengend Snippet: Figure 6. Absence of change in NOX5 density during the development of heart failure in the hereditary cardiomyopathic hamster. (A) Autoradiographs showing the density of NOX5 in NH, HCMH and rat hearts. (B) Bar graph showing that the NOX5/GAPDH ratio does not change in HCMH hearts compared to those from age-matched NHs. This ratio in rat hearts is not different from that in NH hearts. The values are expressed as mean ± SEM where n is the number of Western blot wells of at least three different animals.

Article Snippet: In order to confirm equal loading of proteins, the blots were also probed with a mouse monoclonal antibody against GAPDH (6C5) (Abcam, Ontario, Canada).

Techniques: Western Blot