Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Insights into the Antibacterial Properties of Complement Peptides C3a, C4a, and C5a across Vertebrates.

doi: 10.4049/jimmunol.2101019

Figure Lengend Snippet: FIGURE 1. Antibacterial properties of vertebrate C3a, C4a, and C5a against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1. (AC) Antibacterial activities of M. musculus (mm) C3a, C4a, and C5a against E. coli ATCC25922 (A), P. aeruginosa ATCC 9027 (B), and E. ictaluri HSN-1 (C). (DF) Antibacterial activities of the overlapping peptides derived from C3a, C4a, and C5a of representative vertebrates against E. coli ATCC25922 (D), P. aeruginosa ATCC 9027 (E), and E. ictaluri HSN-1 (F). The underlay agarose gel containing E. coli ATCC25922, P. aeruginosa ATCC 9027, or E. ictaluri HSN-1 was poured into petri dishes. After solidification, 4-mm-diameter wells were punched in the agarose gel, and 6 ml of water (blank control [Ctrl]), LL-37, CATH-1d, M. musculus C3a, C4a, and C5a (120 mM), and overlapping peptides (125 mM) were added to each well. Plates were incubated at 37◦C (for E. coli and P. aeruginosa) or 28◦C (for E. ictaluri) for 3 h, and then the underlay agarose gel was covered with overlay agarose gel. Antibacterial activity of a peptide was measured as a zone of clearing around each well after 20 h of incubation. Data shown in (A)(C) are the means ± SEM of four reactions in a repre- sentative of two independent experiments, whereas those in (D)(F) are representative results of two independent experiments.

Article Snippet: Antibacterial properties of vertebrate C3a, C4a, and C5a To explore the antibacterial properties and structure function relationships of C3a, C4a, and C5a in vertebrates, we synthesized and tested the intact C3a, C4a, and C5a of M. musculus and overlapping peptides (29 35 aa residues) covering C3a, C4a, and C5a of representative vertebrates (Supplemental Table II) for antibacterial activities against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1.

Techniques: Derivative Assay, Agarose Gel Electrophoresis, Control, Incubation, Activity Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Insights into the Antibacterial Properties of Complement Peptides C3a, C4a, and C5a across Vertebrates.

doi: 10.4049/jimmunol.2101019

Figure Lengend Snippet: FIGURE 2. Dose-dependent antibacterial activities of representative peptides against E. coli ATCC25922. (A) Antibacterial activities of peptides against different concentrations of E. coli ATCC25922. The underlay agarose gel containing different concentrations of E. coli ATCC25922, from 8 × 105 to 8 × 108 CFUs/ml, was poured into petri dishes. After solidification, 4-mm-diameter wells were punched in the agarose gel, and 6 ml of peptides (125 mM) was added to each well. Plates were incubated at 37◦C for 3 h, and then the underlay agarose gel was covered with overlay agarose gel. Antibacterial activity of a peptide was measured as a zone of clearing around each well after 20 h of incubation. (B) Antibacterial activities of different concentrations of peptides against E. coli ATCC25922. The underlay agarose gel containing E. coli ATCC25922 (8 × 105 CFUs/ml) was poured into petri dishes. After solidification, 4-mm-diameter wells were punched in the agarose gel, and 6 ml of different concentrations of peptides, from 4 to 128 mM, was added to each well. Plates were incubated at 37◦C for 3 h, and then the underlay agarose gel was covered with overlay agarose gel. Antibacterial activity of a peptide was measured as a zone of clearing around each well after 20 h of incubation. Data shown are the means ± SEM of three reactions in a representative of two independent experiments. Different letters indicate significant differences (p < 0.05).

Article Snippet: Antibacterial properties of vertebrate C3a, C4a, and C5a To explore the antibacterial properties and structure function relationships of C3a, C4a, and C5a in vertebrates, we synthesized and tested the intact C3a, C4a, and C5a of M. musculus and overlapping peptides (29 35 aa residues) covering C3a, C4a, and C5a of representative vertebrates (Supplemental Table II) for antibacterial activities against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1.

Techniques: Agarose Gel Electrophoresis, Incubation, Activity Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Insights into the Antibacterial Properties of Complement Peptides C3a, C4a, and C5a across Vertebrates.

doi: 10.4049/jimmunol.2101019

Figure Lengend Snippet: FIGURE 3. Antibacterial mechanisms of vertebrate C3a-, C4a-, and C5a-derived peptides (A) Bacterial membrane permeability of representative overlapping peptides derived from M. musculus (mm) or C. milii (cm) C3a, C4a, or C5a assessed by flow cytometry. E. coli ATCC25922 were washed and resuspended in NaPB, and then peptides were added to give a final concentration of 8 mM. In addition, a bacterial suspen- sion without peptide was included as a control (Ctrl). After being incubated at 37◦C for 1 h, PI was added to the concentration of 9 mM. The influx of PI into bacterial cells was investigated by flow cytometry at 3000 events. The cell-penetrating efficiency was analyzed using the FlowJo software v10 (Tree Star). (B) Observation of bacteria treated with representa- tive overlapping peptides derived from M. musculus or C. milii C3a, C4a, or C5a by SEM. E. coli ATCC25922 were washed and resuspended in NaPB, and then peptides were added to give a final concen- tration of 8 mM. In addition, a bacterial suspension without peptide was included as a control (Ctrl). After being incubated at 37◦C for 2 h, the bacteria were fixed, dehydrated, vacuum dried, sputter coated with gold, and observed with a field emission SEM at an accelerating voltage of 3 kV. Data shown in (A) and (B) are representative results of two indepen- dent experiments.

Article Snippet: Antibacterial properties of vertebrate C3a, C4a, and C5a To explore the antibacterial properties and structure function relationships of C3a, C4a, and C5a in vertebrates, we synthesized and tested the intact C3a, C4a, and C5a of M. musculus and overlapping peptides (29 35 aa residues) covering C3a, C4a, and C5a of representative vertebrates (Supplemental Table II) for antibacterial activities against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1.

Techniques: Derivative Assay, Membrane, Permeability, Flow Cytometry, Concentration Assay, Control, Incubation, Software, Bacteria, Suspension

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Insights into the Antibacterial Properties of Complement Peptides C3a, C4a, and C5a across Vertebrates.

doi: 10.4049/jimmunol.2101019

Figure Lengend Snippet: FIGURE 7. Antibacterial activities of the mutant peptides against E. coli ATCC25922. (A) Mutants of antibacterial overlapping peptides derived from M. musculus or C. milii C3a, C4a, or C5a. (B) Mutants of inactive overlapping peptides derived from H. sapiens or C. milii C5a. (C) Mutants of H. sapiens LL-37 and O. mykiss CATH-1d. The positively and negatively charged residues are shown in red and blue, respectively, and the mutated residues are shaded in gray. (D) Antibacterial activities of the peptides in (A) against E. coli ATCC25922. (E) Antibacterial activities of the peptides in (B) against E. coli ATCC25922. (F) Antibacterial activities of the peptides in (C) against E. coli ATCC25922. The underlay agarose gel containing E. coli ATCC25922 was poured into petri dishes. After solidification, 4-mm- diameter wells were punched in the agarose gel, and 6 ml of peptides (125 mM) was added to each well. Plates were incubated at 37◦C for 3 h, and then the underlay agarose gel was covered with overlay agarose gel. Antibacterial activity of a peptide was measured as a zone of clearing around each well after 20 h of incubation. Data shown are the means ± SEM of two independent experiments. Different letters indicate significant differences (p < 0.05). DE-R, Asp and Glu to Arg mutant; DE-S, Asp and Glu to Ser mutant; E-R, Glu to Arg mutant; R-S, Arg to Ser mutant; Scrm, scrambled mutant; S-R, Ser to Arg mutant; WT, wild type.

Article Snippet: Antibacterial properties of vertebrate C3a, C4a, and C5a To explore the antibacterial properties and structure function relationships of C3a, C4a, and C5a in vertebrates, we synthesized and tested the intact C3a, C4a, and C5a of M. musculus and overlapping peptides (29 35 aa residues) covering C3a, C4a, and C5a of representative vertebrates (Supplemental Table II) for antibacterial activities against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1.

Techniques: Mutagenesis, Derivative Assay, Agarose Gel Electrophoresis, Incubation, Activity Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Insights into the Antibacterial Properties of Complement Peptides C3a, C4a, and C5a across Vertebrates.

doi: 10.4049/jimmunol.2101019

Figure Lengend Snippet: FIGURE 8. Antibacterial activities of the small peptides consisting of 1011 aa residues derived from highly charged fish C3a, C4a, or C5a. (A) The sequences of small peptides derived from C. milii (cm) C3a and C4a, as well as D. rerio (dr) C5a. The positively charged residues are shown in red, and the negatively charged residues are shown in blue. (B) Antibacterial activities of the small peptides against E. coli ATCC25922. The underlay aga- rose gel containing E. coli ATCC25922 was poured into petri dishes. After solidification, 4-mm-diameter wells were punched in the agarose gel, and 6 ml of peptides (125 mM) was added to each well. Plates were incubated at 37◦C for 3 h, and then the underlay agarose gel was covered with overlay agarose gel. Antibacterial activity of a peptide was measured as a zone of clearing around each well after 20 h of incubation at 37◦C. (C) Bacterial membrane permeability of two representative small peptides assessed by flow cytometry. E. coli ATCC25922 were washed and resuspended in NaPB, and then peptides were added to give a final concentration of 8 mM. In addition, a bacterial suspension without peptide was included as a control (Ctrl). After incubation at 37◦C for 1 h, PI was added to give a final concentration of 9 mM. The influx of PI into bacterial cells was investigated by flow cytometry at 3000 events. The cell-penetrating efficiency was analyzed using the FlowJo software v10 (Tree Star). (D) Observation of bacteria treated with two representative small peptides by SEM. E. coli ATCC25922 were washed and resuspended in NaPB, and then peptides were added to give a final concentration of 8 mM. In addition, a bacterial suspension without peptide was included as a control (Ctrl). After incubation at 37◦C for 2 h, the bacteria were fixed, dehydrated, vacuum dried, sputter coated with gold, and observed with a field emission SEM at an accelerating voltage of 3 kV. Data shown in (B) are the means ± SEM of two independent experiments, whereas those in (C) and (D) are representative results of two indepen- dent experiments.

Article Snippet: Antibacterial properties of vertebrate C3a, C4a, and C5a To explore the antibacterial properties and structure function relationships of C3a, C4a, and C5a in vertebrates, we synthesized and tested the intact C3a, C4a, and C5a of M. musculus and overlapping peptides (29 35 aa residues) covering C3a, C4a, and C5a of representative vertebrates (Supplemental Table II) for antibacterial activities against E. coli ATCC25922, P. aeruginosa ATCC 9027, and E. ictaluri HSN-1.

Techniques: Derivative Assay, Agarose Gel Electrophoresis, Incubation, Activity Assay, Membrane, Permeability, Flow Cytometry, Concentration Assay, Suspension, Control, Software, Bacteria

Journal: Biotechnology Reports

Article Title: Nanotechnology: The new perspective in precision agriculture

doi: 10.1016/j.btre.2017.03.002

Figure Lengend Snippet: Nanofertilizers and their action against plant pathogens.

Article Snippet: The synthesized zinc oxide nanoparticles were evaluated for antibacterial activity against gram negative bacteria Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 15442), gram positive Staphylococcus aureus (ATCC 6538) and Bacillus thuringiensis (ATCC 10792).

Techniques:

Journal: Biotechnology Reports

Article Title: Nanotechnology: The new perspective in precision agriculture

doi: 10.1016/j.btre.2017.03.002

Figure Lengend Snippet: Nanoparticles used as fungicides against plant pathogenic fungi.

Article Snippet: The synthesized zinc oxide nanoparticles were evaluated for antibacterial activity against gram negative bacteria Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 15442), gram positive Staphylococcus aureus (ATCC 6538) and Bacillus thuringiensis (ATCC 10792).

Techniques: