Pointprobe NCSTR

Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction

The emergence of multiple antibiotic-resistant bacteria, notably, pan-resistant Gram-negative pathogens, which are equipped with an outer membrane barrier of low permeability to antibiotics, has become an important challenge in recent decades following the overuse of antibiotics in humans and animals. *

In particular, the foodborne enteric pathogen Escherichia coli O157:H7 has caused severe or deadly illness cases worldwide. *

Among E. coli O157 isolates, serotype O157:H7 is the most common enteric pathogen isolated from patients with bloody diarrhea and it is also frequently found in non-bloody diarrhea samples. Many of its clinical isolates from humans and animals as well as isolates from contaminated food have been found to develop resistance to several antibiotics. *

Following the first isolation of mastoparan, the most abundant peptide in the hornet or wasp venom, from Vespula lewisii, many homologs of mastoparan were isolated from various hornets and solitary wasps. *

Mastoparan homologs are cationic tetradecapeptides with membrane permeabilizing activity and antimicrobial activity on various bacteria, mast cell degranulation activity, and hemolytic activity. *

In the article “Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction” Chun-Hsien Lin, Ching-Lin Shyu, Zong-Yen Wu, Chao-Min Wang, Shiow-Her Chiou, Jiann-Yeu Chen, Shu-Ying Tseng, Ting-Er Lin, Yi-Po Yuan, Shu-Peng Ho, Kwong-Chung Tung, Frank Chiahung Mao, Han-Jung Lee and Wu-chun Tu investigate the antimicrobial activity and membrane disruption modes of the antimicrobial peptide mastoparan-AF against hemolytic Escherichia coli O157:H7.*

Based on the physicochemical properties, mastoparan-AF may potentially adopt a 3–11 amphipathic helix-type structure, with five to seven nonpolar or hydrophobic amino acid residues forming the hydrophobic face. E. coli O157:H7 and two diarrheagenic E. coli veterinary clinical isolates, which are highly resistant to multiple antibiotics, are sensitive to mastoparan-AF, with minimum inhibitory and bactericidal concentrations (MIC and MBC) ranging from 16 to 32 μg mL−1 for E. coli O157:H7 and four to eight μg mL−1 for the latter two isolates. *

Mastoparan-AF treatment, which correlates proportionally with membrane permeabilization of the bacteria, may lead to abnormal dents, large perforations or full opening at apical ends (hollow tubes), vesicle budding, and membrane corrugation and invagination forming irregular pits or pores on E. coli O157:H7 surface. In addition, mRNAs of prepromastoparan-AF and prepromastoparan-B share a 5′-poly(A) leader sequence at the 5′-UTR known for the advantage in cap-independent translation. *

This is the first report about the physicochemical adaptation of 3–11 amphipathic helices among mastoparans or antimicrobial peptides. *

Considering that E. coli O157:H7 and clinical isolates are highly resistant to multiple classes of antibiotics, mastoparan-AF, with little or mild effect on animal RBCs, could be an effective and alternative treatment to combat hemolytic E. coli O157:H7 and other pathogenic E. coli.*

The topography of bacteria was measured by a commercial atomic force microscope using NanoWorld Pointprobe® NCSTR AFM probes with a typical resonance frequency of 160 kHz and a typical spring constant of 7.4 N/m, respectively. For image quality, the scan rates of the tip were 0.3–0.6 Hz, with a resolution set of 512 by 256 pixels, and the feedback control parameters were optimized. *

Figure 5 from «Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction» by Chun-Hsien Lin et al.:The topology of mastoparan-AF treated-hemolytic E. coli O157:H7 analyzed by AFM. (A) Two-dimensional (2D) and (B) three-dimensional (3D) images show smooth cell surfaces of untreated hemolytic E. coli O157:H7. (C) A 2D image of mastoparan-AF (32 μg mL−1)-treated hemolytic E. coli O157:H7. Abnormal perforations and dents on the surface of bacteria are indicated by arrows and arrowheads, respectively. The 3D images focusing on two highlighted areas of (C), respectively, reveal (D) a rough cell surface and (E) a hollow tube resulting from perforations at apical ends. (F) A 3D image shows a mastoparan-AF-treated bacterium with a budding vesicle. (G) A 3D image shows mastoparan-AF-treated bacteria with a wrinkled or rough surface. (H) Magnification of portion of (G) displays, in high resolution, the surface roughness of a mastoparan-AF-treated bacterium. The topography of bacteria was measured by a commercial atomic force microscope using NanoWorld Pointprobe® NCSTR AFM probes with a typical resonance frequency of 160 kHz and a typical spring constant of 7.4 N/m, respectively. For image quality, the scan rates of the tip were 0.3–0.6 Hz, with a resolution set of 512 by 256 pixels, and the feedback control parameters were optimized. * — Figure 5 from «Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction» by Chun-Hsien Lin et al.:
The topology of mastoparan-AF treated-hemolytic E. coli O157:H7 analyzed by AFM. (A) Two-dimensional (2D) and (B) three-dimensional (3D) images show smooth cell surfaces of untreated hemolytic E. coli O157:H7. (C) A 2D image of mastoparan-AF (32 μg mL−1)-treated hemolytic E. coli O157:H7. Abnormal perforations and dents on the surface of bacteria are indicated by arrows and arrowheads, respectively. The 3D images focusing on two highlighted areas of (C), respectively, reveal (D) a rough cell surface and (E) a hollow tube resulting from perforations at apical ends. (F) A 3D image shows a mastoparan-AF-treated bacterium with a budding vesicle. (G) A 3D image shows mastoparan-AF-treated bacteria with a wrinkled or rough surface. (H) Magnification of portion of (G) displays, in high resolution, the surface roughness of a mastoparan-AF-treated bacterium.

*Chun-Hsien Lin, Ching-Lin Shyu, Zong-Yen Wu, Chao-Min Wang, Shiow-Her Chiou, Jiann-Yeu Chen, Shu-Ying Tseng, Ting-Er Lin, Yi-Po Yuan, Shu-Peng Ho, Kwong-Chung Tung, Frank Chiahung Mao, Han-Jung Lee and Wu-chun Tu
Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction
Membranes 2023, 13(2), 251
DOI: https://doi.org/10.3390/membranes13020251

The article “Antimicrobial Peptide Mastoparan-AF Kills Multi-Antibiotic Resistant Escherichia coli O157:H7 via Multiple Membrane Disruption Patterns and Likely by Adopting 3–11 Amphipathic Helices to Favor Membrane Interaction” by Chun-Hsien Lin, Ching-Lin Shyu, Zong-Yen Wu, Chao-Min Wang, Shiow-Her Chiou, Jiann-Yeu Chen, Shu-Ying Tseng, Ting-Er Lin, Yi-Po Yuan, Shu-Peng Ho, Kwong-Chung Tung, Frank Chiahung Mao, Han-Jung Lee and Wu-chun Tu is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.

Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces

Poly(3-hexylthiophene) (P3HT), a hole-conducting polymer, generates a lot of interest especially because of its excellent optoelectronic properties (such as good electrical conductivity and high extinction coefficient) and good processability, which make this polymer an excellent choice for building organic optoelectronic devices (e.g., organic solar cells). *

P3HT films and nanoparticles have also been used to restore the photosensitivity of retinal neurons. *

For their article “Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces” Szilveszter Gáspár, Tiziana Ravasenga, Raluca-Elena Munteanu, Sorin David, Fabio Benfenati, and Elisabetta Colombo investigated the template-assisted electrochemical synthesis of P3HT nanowires doped with tetrabutylammonium hexafluorophosphate (TBAHFP) and their biocompatibility with primary neurons. *

They were able to show that template-assisted electrochemical synthesis can relatively easily turn 3-hexylthiophene (3HT) into longer (e.g., 17 ± 3 µm) or shorter (e.g., 1.5 ± 0.4 µm) P3HT nanowires with an average diameter of 196 ± 55 nm (determined by the used template) and that the nanowires produce measurable photocurrents following illumination. *

The fact that template-assisted electrochemical synthesis combines polymerization, doping, and polymer nanostructuring into one, relatively simple step is the most important advantage of this method. The possibility of easily tuning the length of the produced nanowires represents another important advantage. *

The authors were also able to demonstrate that primary cortical neurons can be grown onto P3HT nanowires drop-casted on a glass substrate without relevant changes in their viability and electrophysiological properties, indicating that P3HT nanowires obtained by template-assisted electrochemical synthesis represent a promising neuronal interface for photostimulation. *

Szilveszter Gáspár et al. proved the biocompability of the obtained P3HT nanowires upon incubation for different periods with primary neuronal cultures. They demonstrated that their presence does not affect the membrane properties of the neurons or the excitability of the neurons as evaluated by patch-clamp experiments. These results show the potential of the described synthesis methodology to fabricate injectable P3HT-based photosensitive nanowires with high biocompatibility, ultimately paving the way for their exploitation for neuronal photostimulation. *

Atomic Force Microscopy (AFM) was used to characterize P3HT nanowires drop-casted onto glass coverslips. *

The Atomic Force Microscopy images were obtained in air and in intermittent contact-mode using line rates as slow as 0.2 Hz and NanoWorld Pointprobe® NCSTR silicon soft-tapping AFM probes (typical values: resonant frequency 160 kHz, force constant 7.2 N m). The ratio between the set-point amplitude and the free amplitude of the AFM cantilever was set to 0.5–0.6. The obtained AFM images were used to determine both the lengths and the diameters of the nanowires. *

Figure 3 from “Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces” by Szilveszter Gáspár et al.: AFM images of “long” P3HT nanowires (A) and of “short” P3HT nanowires (B). NanoWorld Pointprobe NCSTR soft-tapping mode probes were used. — Figure 3 from “Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces” by Szilveszter Gáspár et al.:
AFM images of “long” P3HT nanowires (A) and of “short” P3HT nanowires (B).

*Szilveszter Gáspár, Tiziana Ravasenga, Raluca-Elena Munteanu, Sorin David, Fabio Benfenati, and Elisabetta Colombo
Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces
Materials 2021, 14(16), 4761, Special Issue Advanced Designs of Materials, Devices and Techniques for Biosensing
DOI: https://doi.org/10.3390/ma14164761 (please follow this external link to read the full article.)

Open Access The article “Electrochemically Synthesized Poly(3-hexylthiophene) Nanowires as Photosensitive Neuronal Interfaces” by Szilveszter Gáspár, Tiziana Ravasenga, Raluca-Elena Munteanu, Sorin David, Fabio Benfenati, and Elisabetta Colombo is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

A Short Peptide Hydrogel with High Stiffness Induced by 310‐Helices to β‐Sheet Transition in Water

In the article “A Short Peptide Hydrogel with High Stiffness Induced by 3₁₀‐Helices to β‐Sheet Transition in Water” by Shu Hui Hiew, Harini Mohanram, Lulu Ning, Jingjing Guo, Antoni Sánchez‐Ferrer, Xiangyan Shi, Konstantin Pervushin, Yuguang Mu, Raffaele Mezzenga and Ali Miserez, a short biomimetic peptide composed of eight amino acid residues derived from squid sucker ring teeth proteins is demonstrated to form hydrogel in water without any cross‐linking agent or chemical modification and exhibits a stiffness on par with the stiffest peptide hydrogels.
Their study broadens the range of secondary structures available to create supramolecular hydrogels, and introduces 3_10‐helices as transient building blocks for gelation via a 3_10‐to‐β‐sheet conformational transition.*

The AFM images presented in this study were obtained in soft tapping mode using NanoWorld Pointprobe® NCSTR AFM probes.

*Shu Hui Hiew, Harini Mohanram, Lulu Ning, Jingjing Guo, Antoni Sánchez‐Ferrer, Xiangyan Shi, Konstantin Pervushin, Yuguang Mu, Raffaele Mezzenga, Ali Miserez
A Short Peptide Hydrogel with High Stiffness Induced by 3_10‐Helices to β‐Sheet Transition in Water
Advanced Science 2019, 1901173
Doi: https://doi.org/10.1002/advs.201901173

Please follow this external link to read the full article: https://onlinelibrary.wiley.com/doi/full/10.1002/advs.201901173

Open Access: The article « A Short Peptide Hydrogel with High Stiffness Induced by 3₁₀‐Helices to β‐Sheet Transition in Water » ” by Shu Hui Hiew, Harini Mohanram, Lulu Ning, Jingjing Guo, Antoni Sánchez‐Ferrer, Xiangyan Shi, Konstantin Pervushin, Yuguang Mu, Raffaele Mezzenga and Ali Miserez is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other thirdparty material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.