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Currently, Ag/AgCl-based reference electrodes are used in most electrochemical biosensors and other bioelectrochemical devices. However, standard reference electrodes are rather large and do not always fit within electrochemical cells designed for the determination of analytes in low-volume aliquots. Therefore, various designs and improvements in reference electrodes are critical for the future development of electrochemical biosensors and other bioelectrochemical devices. In this study, we explain a procedure to apply common laboratory polyacrylamide hydrogel in a semipermeable junction membrane between the Ag/AgCl reference electrode and the electrochemical cell. During this research, we have created disposable, easily scalable, and reproducible membranes suitable for the design of reference electrodes. Thus, we came up with castable semipermeable membranes for reference electrodes. Performed experiments highlighted the most suitable gel formation conditions to achieve optimal porosity. Here, Cl− ion diffusion through the designed polymeric junctions was evaluated. The designed reference electrode was also tested in a three-electrode flow system. The results show that home-built electrodes can compete with commercial products due to low reference electrode potential deviation (~3 mV), long shelf-life (up to six months), good stability, low cost, and disposability. The results show a high response rate, which makes in-house formed polyacrylamide gel junctions good membrane alternatives in the design of reference electrodes, especially for these applications where high-intensity dyes or toxic compounds are used and therefore disposable electrodes are required.

Prolonged storage of frozen dough worsens the structure of thawed dough. The main reason is the inhibition of yeast activity. In this study we investigated applicability of pulsed electric field treatment for introduction of cryoprotectant into yeast cells. We showed that pre-treatment of cells suspended in a trehalose solution improves freezing tolerance and results in higher viability after thawing. Viability increased with rise in electric field strength (from 3 to 4.5 kV/cm) and incubation time (from 0 to 60 min) after exposure. Pretreatment resulted in lower decrease in the viability of thawed cells, viability of untreated cells dropped to 10%, while pre-treatment with PEF and trehalose tripled the viability.

The design and development of a compact square-wave pulse generator for the electroporation of biological cells is presented. This electroporator can generate square-wave pulses with durations from 3 μs up to 10 ms, voltage amplitudes up to 3500 V, and currents up to 250 A. The quantity of the accumulated energy is optimized by means of a variable capacitor bank. The pulse forming unit design uses a crowbar circuit, which gives better control of the pulse form and its duration, independent of the load impedance. In such cases, the square-wave pulse form ensures better control of electroporation efficiency by choosing parameters determined in advance. The device has an integrated graphic LCD screen and measurement modules for the visualization of the current pulse, allowing for express control of the electroporation quality and does not require an external oscilloscope for current pulse recording. This electroporator was tested on suspensions of Saccharomyces cerevisiae yeast cells, during which, it was demonstrated that the application of such square-wave pulses ensured better control of the electroporation efficiency and cell viability after treatment using the pulsed electric field (PEF).

Abstract Pulsed electric field (PEF) treatment is known to cause plasma membrane permeabilization of microorganisms, an effect known as electroporation. PEF treatment is very attractive since it can achieve permeabilization with or without lethal damage in accordance with desired results. This study aimed to expand the accomplishment of electroporation outcomes by applying sudden post-PEF osmotic composition change of the media. Changes in yeast cells’ viability, size and plasma membrane regeneration rate were evaluated. However, we still have questions about the intracellular biochemical processes responsible for plasma membrane recovery after electroporation. Our suggested candidate is the high osmolarity glycerol (HOG) kinase pathway. The HOG pathway in Saccharomyces cerevisiae yeasts is responsible for volume recovery after dangerous shape modifications and intracellular water disbalance caused by environmental osmotic pressure changes. Thus, we evaluated the HOG pathway inactivation effect on S. cerevisiae’s reaction to PEF treatment. Results showed that Hog1 deficient S. cerevisiae cells were considerably more sensitive to electric field treatment, confirming a link between the HOG pathway and S. cerevisiae recovery process after electroporation. By suddenly changing the osmolarity of the media after PEF we influenced the cells’ plasma membrane recovery rate, severity of permeabilization and survivability of yeast cells. Studies of electroporation in combination with various treatments might improve electric field application range, efficiency, and optimization of the process.

Pulsed electric field (PEF) as a green processing technology is drawing greater attention due to its eco-friendliness and potential to promote sustainable development goals. In this study, the effects of different electric field strengths (EFS, 0–30 kV/cm) on the structure and physicochemical features of casein micelles (CSMs) were investigated. It was found that the particle sizes of CSMs increased at low EFS (10 kV/cm) but decreased at high EFS (30 kV/cm). The absolute ζ-potential at 30 kV/cm increased from −26.6 (native CSMs) to −29.5 mV. Moreover, it was noticed that PEF treatment leads to changes in the surface hydrophobicity; it slightly increased at low EFS (10 kV/cm) but decreased at EFS > 10 kV/cm. PEF enhanced the protein solubility from 84.9 (native CSMs) to 87.1% (at 10 kV/cm). PEF at low EFS (10 kV/cm) intensified the emission fluorescence spectrum of CSMs, while higher EFS reduced the fluorescence intensity compared to native CSMs. Moreover, the analysis of the Amide Ι region showed that PEF-treated CSMs reduced the α-helix and increased the β-sheet content. Raman spectra confirmed that PEF treatment > 10 kV/cm buried tyrosine (Tyr) residues in a hydrophobic environment. It was also found that PEF treatment mainly induced changes in the disulfide linkages. In conclusion, PEF technology can be employed as an eco-friendly technology to change the structure and physiochemical properties of CSMs; this could improve their techno-functional properties.

Currently, microbial biofilms have been the cause of a wide variety of infections in the human body, reaching 80% of all bacterial and fungal infections. The biofilms present specific properties that increase the resistance to antimicrobial treatments. Thus, the development of new approaches is urgent, and antimicrobial photodynamic therapy (aPDT) has been shown as a promising candidate. aPDT involves a synergic association of a photosensitizer (PS), molecular oxygen and visible light, producing highly reactive oxygen species (ROS) that cause the oxidation of several cellular components. This therapy attacks many components of the biofilm, including proteins, lipids, and nucleic acids present within the biofilm matrix; causing inhibition even in the cells that are inside the extracellular polymeric substance (EPS). Recent advances in designing new PSs to increase the production of ROS and the combination of aPDT with other therapies, especially pulsed electric fields (PEF), have contributed to enhanced biofilm inhibition. The PEF has proven to have antimicrobial effect once it is known that extensive chemical reactions occur when electric fields are applied. This type of treatment kills microorganisms not only due to membrane rupture but also due to the formation of reactive compounds including free oxygen, hydrogen, hydroxyl and hydroperoxyl radicals. So, this review aims to show the progress of aPDT and PEF against the biofilms, suggesting that the association of both methods can potentiate their effects and overcome biofilm infections.

Dairy and plant-based proteins are widely utilized in various food applications. Several techniques have been employed to improve the techno-functional properties of these proteins. Among them, pulsed electric field (PEF) technology has recently attracted considerable attention as a green technology to enhance the functional properties of food proteins. In this review, we briefly explain the fundamentals of PEF devices, their components, and pulse generation and discuss the impacts of PEF treatment on the structure of dairy and plant proteins. In addition, we cover the PEF-induced changes in the techno-functional properties of proteins (including solubility, gelling, emulsifying, and foaming properties). In this work, we also discuss the main challenges and the possible future trends of PEF applications in the food proteins industry. PEF treatments at high strengths could change the structure of proteins. The PEF treatment conditions markedly affect the treatment results with respect to proteins’ structure and techno-functional properties. Moreover, increasing the electric field strength could enhance the emulsifying properties of proteins and protein-polysaccharide complexes. However, more research and academia–industry collaboration are recommended to build highly effective PEF devices with controlled processing conditions.