Journal: Advanced Science

Article Title: Utilizing Proteolytic‐Resistant Nano‐Short Peptide Based on Naphthyl Tail‐Anchored to Combat Bacterial Infections

doi: 10.1002/advs.202508854

Figure Lengend Snippet: In vitro barrier penetration and antimicrobial properties of nano‐short peptides. A) The minimum inhibitory concentration (MIC) values of nano‐short peptides against E. coli ATCC 25922 in the presence of physiological salts and serum. A value of 256 indicates no detectable antimicrobial activity under the test conditions ( n = 3). B) MIC values of nano‐short peptides against E. coli ATCC 25922 after incubation for 1 h with different concentrations (1, 2, 4, 8 mg mL −1 ) of various proteases. A value of 256 indicates no detectable antimicrobial activity under the test conditions ( n = 3). C) Cleavage of N 4 by various proteases. The peptides were incubated with protease for 0, 1, 2, 4, and 8 h at 37 °C, and the molecular weights of the protein markers from top to bottom were 31, 14.4, 7.8, 5.8 and 3.3 kDa. D) RP‐HPLC analysis of N 4 after incubation with various proteases at a final concentration of 8 mg mL −1 for 0, 1, 2, 4, and 8 h. E) Time‐kill kinetic curves of E. coli ATCC 25922 after treatment with different concentrations of N 4 . Data are the mean ± SD; n = 3. F) The drug resistance curves of E . coli ATCC 25922 to N 4 and gentamicin during a 30‐day continuous induction period. G) MIC values of N 4 and antibiotics against clinical strains. A value of 128 indicates no detectable antimicrobial activity at a concentration of 64 × 10 −6 m , a value of 1 indicates the antimicrobial activity is less than 2 × 10 −6 m ( n = 3).

Article Snippet: [ ] It is well‐established that the β‐galactosidase‐mediated hydrolysis of o‐nitrophenyl β‐D‐galactopyranoside (ONPG) serves as a quantitative indicator of CM integrity in E. coli ATCC 25922.

Techniques: In Vitro, Concentration Assay, Activity Assay, Incubation

Journal: Advanced Science

Article Title: Utilizing Proteolytic‐Resistant Nano‐Short Peptide Based on Naphthyl Tail‐Anchored to Combat Bacterial Infections

doi: 10.1002/advs.202508854

Figure Lengend Snippet: Antimicrobial mechanism of nano‐short peptides. A) Lipopolysaccharides (LPS) binding affinities of A 4 , B 4 , and N 4 . B) Effect of A 4 , B 4 , and N 4 on the outer membrane permeability of E. coli ATCC 25922. C) Effect of A 4 , B 4 , and N 4 on the cell membrane integrity of E. coli ATCC 25922 at different concentrations. D) Depolarization ability of A 4 , B 4 , and N 4 on cytoplasmic membrane of E. coli ATCC 25922 at different concentrations. E) Inhibition on respiratory chain dehydrogenase activity of E. coli ATCC 25922 by A 4 , B 4 , and N 4 at different concentrations. F) Intracellular ATP content in E. coli ATCC 25922 cells after treatment with different concentrations of A 4 , B 4 , and N 4 . Differences between groups in (A), (B), (E) and (F) were analyzed by one‐way ANOVA followed by Tukey's multiple comparisons tests. Values with different superscripts (a, b, c, and …g) indicate a significant difference ( p < 0.05). Data are the mean ± SD; n = 3. G) Transmission electron microscopy (TEM) negative staining images of E. coli ATCC 25922 treated with N 4 . Scale bars, 500 nm and 1 µm. H) Scanning electron microscopy (SEM) images of E. coli ATCC 25922 treated with N 4 Scale bars, 1µm. I) TEM images of E. coli ATCC 25922 treated with N 4 . Scale bars, 500 nm. J) Live/dead fluorescence imaging of E. coli ATCC 25922 after N 4 treatment at different concentrations. Scale bars, 100 µm. K) The agglutination of E. coli ATCC 25922 in cuvette induced by different concentrations of N 4 . L) Bacterial content of the E. coli ATCC 25922 supernatant in cuvettes treated with different concentrations of N 4 . M) Flow cytometry imaging of E. coli ATCC 25922 after N 4 treatment. N4 was used for detection at a concentration of 16 × 10 −6 m in (G), (H), (I), and (M).

Article Snippet: [ ] It is well‐established that the β‐galactosidase‐mediated hydrolysis of o‐nitrophenyl β‐D‐galactopyranoside (ONPG) serves as a quantitative indicator of CM integrity in E. coli ATCC 25922.

Techniques: Binding Assay, Membrane, Permeability, Inhibition, Activity Assay, Transmission Assay, Electron Microscopy, Negative Staining, Fluorescence, Imaging, Agglutination, Flow Cytometry, Concentration Assay

Journal: Advanced Science

Article Title: Utilizing Proteolytic‐Resistant Nano‐Short Peptide Based on Naphthyl Tail‐Anchored to Combat Bacterial Infections

doi: 10.1002/advs.202508854

Figure Lengend Snippet: N 4 exhibits excellent efficacy in vivo. A) In vivo efficacy assessment experimental workflow. B) Bacterial load of E. coli in the liver, kidney, and spleen following infection with E. coli and subsequent treatment with saline and N 4 . Differences between groups were analyzed using independent t ‐tests, values with different superscripts indicate a significant difference ( p < 0.05). Data are the mean ± SD; n = 8. C) Histopathological H&E staining of the liver, spleen, and kidney. Scale bars, 100 µm. D) Levels of inflammatory factors (IL‐10, IL‐1β, IL‐6, and TNF‐α) in the serum of mice from different treatment groups. Values with different superscripts (a, b, and c) indicate a significant difference ( p < 0.05). Differences between groups were analyzed by one‐way ANOVA followed by Tukey's multiple comparisons tests. Data are the mean ± SD; n = 8. E) Fluorescence images of M1 macrophage biomarker (TNF‐α, CD86) and M2 macrophage biomarker (IL‐10, CD206) in spleen of mice subjected to different treatments. Scale bars, 50 µm.

Article Snippet: [ ] It is well‐established that the β‐galactosidase‐mediated hydrolysis of o‐nitrophenyl β‐D‐galactopyranoside (ONPG) serves as a quantitative indicator of CM integrity in E. coli ATCC 25922.

Techniques: In Vivo, Infection, Saline, Staining, Fluorescence, Biomarker Discovery

Journal:

Article Title: Affinity Purification Strategy to Capture Human Endogenous Proteasome Complexes Diversity and to Identify Proteasome-interacting Proteins * S⃞

doi: 10.1074/mcp.M800193-MCP200

Figure Lengend Snippet: Detection of 20 S core particle and 19 S activators in the purified proteasome preparations. Proteins separated by SDS-PAGE were transferred to a nitrocellulose membrane: 2 μg of commercial 20 S and 26 S proteasome from human erythrocytes as standards (A), 20 μg of total proteins from fractions containing the maximal ChT-like activity (fractions 9 and 10 from purifications without and with formaldehyde cross-linking, respectively) (B), and 10 μg of estimated 20 S proteasome (based on the ChT-like activity measurement) from fractions 8, 9, and 10 from the purification without formaldehyde cross-linking (C). Mouse monoclonal primary antibodies against 19 S proteasome subunits Rpt1 and Rpn12 and rabbit polyclonal antibodies against 20 S core subunits were used for the immunoblot staining. ECL Plex CyDye-conjugated antibodies, goat α-mouse IgG-Cy3 and goat α-rabbit IgG-Cy5, were used as secondary antibodies. The detection was performed using the Typhoon Trio fluorescence scanner at 532 nm excitation and 580 nm emission for the Cy3-conjugated antibody and 633 nm excitation and 670 nm emission for the Cy5-conjugated antibody. Lane M, molecular mass markers.

Article Snippet: ECL Plex CyDye-conjugated antibodies, goat α-mouse IgG-Cy3 (1:2,000) and goat α-rabbit IgG-Cy5 (1:1,250) (GE Healthcare), were used as secondary antibodies.

Techniques: Purification, SDS Page, Activity Assay, Western Blot, Staining, Fluorescence

Journal: Progress in Biomaterials

Article Title: Hydrogel scaffolds with elasticity-mimicking embryonic substrates promote cardiac cellular network formation

doi: 10.1007/s40204-020-00137-0

Figure Lengend Snippet: Shown in a are cells cultured within the 3D gels and on 2D Matrigel controls. The cells were pre-stained with PKH26 (red) to reveal network formation in these cultures. Also, the magnified insets depict the elongated cell morphology within the embryonic gels, which became gradually rounded as the gels were progressively stiffened from the physiologic to the fibrotic gels. Shown in b is the average cellular aspect ratio (width/height) for cells cultured within the gels. The aspect ratio is maximum in the case of cells cultured within the embryonic gels and is close to the value of 1 (implying rounded cells) as the gels were stiffened

Article Snippet: To confirm the viability of cells, post-encapsulation in gels after crosslinking, the CM were labeled with PKH26 red fluorescent membrane staining dye (Sigma), as done before (Anil Kumar et al. 2019a , b ).

Techniques: Cell Culture, Staining

Journal: Cell metabolism

Article Title: Miro1 Marks Parkinson’s Disease Subset and Miro1 Reducer Rescues Neuron Loss in Parkinson’s Models.

doi: 10.1016/j.cmet.2019.08.023

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: For fluorescent Western, blots were probed with Cy5-conjugated goat anti-mouse IgG (PA45009, GE Healthcare) at 1:5,000, and scanned at 635nm with a Molecular Dynamics Storm 860 Imager (Amersham BioSciences, Piscataway, NJ) in a linear range for fluorescent detection.

Techniques: Recombinant, Activity Assay, Software