Safety and effectiveness of nano-pulse stimulation™ technology to treat acne vulgaris of the back.

BACKGROUND AND OBJECTIVES: This feasibility study describes the effects of Nano-pulse stimulation™ (NPS™) technology using the CellFX™ System on acne vulgaris of the back with the objectives of demonstrating safety and effectiveness. The CellFX System applies nanosecond pulses of electrical energy to induce highly localized regulated cell death (RCD) in the cellular structures of the targeted zone with no thermal effect on the tissue and negligible effects on surrounding non-cellular components. STUDY DESIGN/MATERIALS AND METHODS: Seventeen subjects were enrolled at two sites with thirteen subjects completing treatment. Three 7 X 7 cm regions containing at least five bacne lesions each were identified, one region treated with the CellFX across three treatment sessions, the second region treated as a sham using microneedle tip placement without delivering energy, and the third as an untreated control. RESULTS: CellFX-treated areas showed an average reduction of acne lesions of 82% by 90 days post-last procedure. Acne improvement was observed in 100% of CellFX-treated regions compared to 39% improvement in Sham regions and 31% improvement in the control regions. The most common skin effects were erythema and hyperpigmentation observed in 23% and 92% of the subjects, respectively, at the last timepoint. No serious adverse events were reported. CONCLUSIONS: CellFX is a safe and effective procedure for clearing back acne.

Prediction of Aspergillus parasiticus inhibition and aflatoxin mitigation in red pepper flakes treated by pulsed electric field treatment using machine learning and neural networks.

Presence of aflatoxins in agricultural products is a worldwide problem. Because of their high heat stability and resistance to most of the food processing technologies, aflatoxin degradation is still a big challenge. Thus, efficacy of pulsed electric fields (PEF) by energies ranging from 0.97 to 17.28 J was tested to determine changes in quality properties in red pepper flakes, mitigation of aflatoxins, inactivation of aflatoxin producing Aspergillus parasiticus, reduction in aflatoxin mutagenity, and modelling of A. parasiticus inactivation in addition to aflatoxin mitigation. Maximum inactivation rate of 64.37 % with 17.28 J was encountered on the mean initial A. parasiticus count. A 99.88, 99.47, 97.75, and 99.58 % reductions were obtained on the mean initial AfG1, AfG2, AfB1, and AfB2 concentrations. PEF treated samples by 0.97, 1.36, 5.76, and 17.28 J at 1 µg/plate, 0.97, 1.92, 7.78, 10.80 J at 10 µg/plate, and 0.97, 1.92, 2.92, 4.08, 5.76, 4.86, 6.80, 9.60, 10.80, and 10.89 J at 100 µg/plate were not mutagenic. Modelling with gradient boosting regression tree (GBRT), random forest regression (RFR), and artificial neural network (ANN) provided the lowest RMSE and highest R2 value for GBRT model for the predicted inactivation of A. parasiticus, whereas ANN model provided the lowest RMSE and highest R2 for predicted mitigation of AfG1, AfB1, and AfB2. PEF treatment possess a viable alternative for aflatoxin degradation with reduced mutagenity and without adverse effect on quality properties of red pepper flakes.

Pulsed electric field treatment strategies to increase bioaccessibility of phenolic and carotenoid compounds in oil-added carrot purees.

The impact of pulsed electric fields (PEF) and their combination with a thermal treatment on the bioaccessibility of phenolic and carotenoid compounds in oil-added carrot puree (5 %) was investigated. Fractions of such puree were differently treated: subjected to PEF (5 pulses of 3.5 kV cm-1) (PEF); thermally treated (70 °C for 10 min) (T) or first PEF treated and then thermally treated (PEF/T). Purees were in vitro digested, carotenoid and phenolic content and bioaccessibility were determined. Likewise, quality attributes and microstructure were analyzed. Generally, treatments did not affect carotenoid content and quality attributes, whereas phenolic content dramatically decreased after PEF. Nevertheless, all treatments enhanced both compounds bioaccessibilities, which were trebled in PEF-treated purees. Particle size reduction may suggest that microstructural changes could be responsible of bioaccessibility increases. Therefore, PEF could be a feasible treatment to enhance phenolic and carotenoid bioaccessibility without altering quality attributes of carrot-based puree.

Red pitaya extract as natural antioxidant in pork patties with total replacement of animal fat.

The antioxidant effects of red pitaya extract (PE) were evaluated in pork patties for 18 days at 2 °C. The following treatments were prepared: control (CON, without antioxidant), sodium erythorbate (ERY, 500 mg kg-1), PE low dose (PEL, 250 mg kg-1), PE medium dose (PEM, 500 mg kg-1), and PE high dose (PEH, 1000 mg kg-1). No significant effect was observed on chemical composition and cooking loss with the addition of PE, while a significant effect was noticed in cohesiveness (P < 0.05). The intense pink colour of PE enhanced the colour stability during storage (9.33, 7.92 and 7.69 vs. 6.77 for PEH, PEM and PEL vs. CON, respectively; (P < 0.05). TBARS (1.21 vs. 2.44 mg MDA/kg) and carbonyl values (5.45 vs. 6.87 nmol carbonyl/mg) of treated samples were lower than those observed in CON. Similar values were found between samples with PE and ERY. PE improved colour acceptance and the preference of pork patties. Therefore, PE is a very effective natural antioxidant by delaying colour and oxidative deterioration.

A dose-response study of nanosecond electric energy pulses on facial skin.

Nanosecond pulsed electric fields, also known as Nano-Pulse Stimulation or NPS, can trigger regulated cell death to clear skin lesions that are cellular in nature. Before treating facial lesions, it is important to demonstrate the effects of these pulses on normal facial skin. Here we have applied a range of NPS energies to the epidermis and dermis of normal facial skin scheduled for excision to establish a safe dose range of energies prior to use in clinical applications. This was an open-label, non-randomized study under the direction of a single Principal Investigator. The time course of the treated tissue changes was determined by histological analysis. All energy settings generated a delayed epidermal loss followed by re-epithelialization by day 7 and a normal course of healing. One day after NPS treatment, the cellular membranes of the treated epidermis were intact, but their nuclei no longer stained with H&E, resulting in a hollow appearance that has been referred to as "ghost cells." Cellular structures in the dermis, such as sebaceous glands and melanocytes, exhibited regulated cell death observed by 1 day post treatment. Melanocytes recovered to their normal density within 7 days. The 60-day samples indicated that epidermis, hair follicles, and eccrine glands appeared normal. The selective effect of NPS treatment on cellular structures in the epidermal and dermal layers suggests that this non-thermal modality of energy delivery is ideal for treating cellular targets including benign and malignant skin lesions. NPS skin treatments provide a promising method for clearing skin lesions with a cellular basis.

Effect of pulsed electric fields and high pressure on improved recovery of high-added-value compounds from olive pomace.

Olive pomace is considered a solid by-product and a rich source of valuable compounds such as polyphenols, flavonoids with antioxidant properties, and proteins. Nonthermal technologies, which cause alterations to cell permeability, are being explored to assist conventional recovery techniques. The aim of this study was to assess the effect of pulsed electric fields (PEF) and high pressure (HP) on improved recovery yield of the high-added-value compounds or to shorten the extraction time of these compounds. Olive pomace (Tsounati cv) was pretreated with PEF (1.0 to 6.5 kV/cm, 0.9 to 51.1 kJ/kg, and 15 µs pulse width) or HP (200 to 600 MPa and 0 to 40 min). Evaluation of the intracellular compounds extracted via solid-liquid extraction (50% ethanol-water solution) was performed. More intense PEF and HP conditions resulted in a significant increase of the phenolic concentration up to 91.6% and 71.8%, respectively. The increased antioxidant capacity of each extract was correlated to phenolic compound concentration. The protein concentration that was achieved with PEF pretreatment was doubled; however, HP-pretreated extracts reached 88.1% higher yield than untreated for pressures up to 200 MPa. HP and PEF pretreatment decreased extraction completion time t98 (needed time to recover the equal amount of phenolics and proteins of untreated after 60 min of conventional extraction) to 12 min and lower than 1 min, respectively. To conclude, both pretreatments are effective in improving the conventional extraction process for increased yield recovery of high-added-value compounds from olive pomace.

A dose-response study of a novel method of selective tissue modification of cellular structures in the skin with nanosecond pulsed electric fields.

BACKGROUND AND OBJECTIVES: This study describes the effects of nanosecond pulsed electric fields (nsPEF) on the epidermis and dermis of normal skin scheduled for excision in a subsequent abdominoplasty. NsPEF therapy applies nanosecond pulses of electrical energy to induce regulated cell death (RCD) in cellular structures, with negligible thermal effects. Prior pre-clinical studies using nsPEF technology have demonstrated the ability to stimulate a lasting immune response in animal tumor models, including melanoma. This first-in-human-use of nsPEF treatment in a controlled study to evaluate the dose-response effects on normal skin and subcutaneous structures is intended to establish a safe dose range of energies prior to use in clinical applications using nsPEF for non-thermal tissue modification. STUDY DESIGN/MATERIALS AND METHODS: Seven subjects with healthy tissue planned for abdominoplasty excision were enrolled. Five subjects were evaluated in a longitudinal, 60-day study of effects with doses of six nsPEF energy levels. A total of 30 squares of spot sizes 25mm2 or less within the planned excision area were treated and then evaluated at 1 day, 5 days, 15 days, 30 days, and 60 days prior to surgery. Photographs were taken over time of each treated area and assessed by three independent and blinded dermatologists for erythema, flaking and crusting using a 5-point scale (0 = low, 4 = high). Punch biopsies of surgically removed tissue were processed and evaluated for tissue changes using hematoxylin and eosin, trichome, caspase-3, microphthalmia transcription factor, and elastin stains and evaluated by a dermatopathologist. The skin of two subjects received additional treatments at 2 and 4 hours post-nsPEF and was evaluated in a similar manner. RESULTS: Most energy settings exhibited delayed epidermal loss followed by re-epithelization by day 15 and a normal course of healing. Histologic analysis identified the appearance of activated caspase-3 at two and four hours after nsPEF treatment, but not at later time points. At the 1-day time point, a nucleolysis effect was observed in epidermal cells, as evidenced by the lack of nuclear staining while the epidermal plasma membranes were still intact. Cellular structures within the treatment zone such as melanocytes, sebaceous glands, and hair follicles were damaged while acellular structures such as elastic fibers and collagen were largely unaffected except for TL6 which showed signs of dermal damage. Melanocytes reappeared at levels comparable with untreated controls within 1 month of nsPEF treatment. CONCLUSIONS: The selective effect of nsPEF treatment on cellular structures in the epidermal and dermal layers suggests that this non-thermal mechanism for targeting cellular structures does not affect the integrity of dermal tissue within a range of energy levels. The specificity of effects and a favorable healing response makes nsPEF ideal for treating cellular targets in the epidermal or dermal layers of the skin, including treatment of benign and malignant lesions. NsPEF skin treatments provide a promising, non-thermal method for treating skin conditions and removing epidermal lesions. © 2019 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Dual-function of Baicalin in nsPEFs-treated Hepatocytes and Hepatocellular Carcinoma cells for Different Death Pathway and Mitochondrial Response.

Nanosecond pulsed electric fields (nsPEFs) is emerged as a potential curative modality to ablate hepatocellular carcinoma (HCC). The application of local ablation is usually limited by insufficiency of liver function. While baicalin, a flavonoid isolated from Scutellaria baicalensis Georgi, has been proven to possess both anti-tumor and protective effects. Our study aimed to estimate different responses of hepatic cancer cells and hepatocytes to the combination of nsPEFs and baicalin. Cell viability, apoptosis and necrosis, mitochondrial transmembrane potential (MTP) and reactive oxygen species (ROS) were examined by CCK-8, FCM, JC-1 and fluorescent probe, respectively. After treatment by nsPEFs, most hepatocytes died by apoptosis, nevertheless, nearly all cancer cells were killed through necrosis. Low concentration of baicalin synergically enhanced nsPEFs-induced suppression and necrosis of HCC cells, nevertheless, the application of baicalin protected normal hepatocytes from the injury caused by nsPEFs, owing to elevating mitochondrial transmembrane potential and reducing ROS generation. Our work provided an advantageous therapy for HCC through the enhanced combination treatment of nsPEFs and baicalin, with which could improve the tumor-ablation effect and alleviate the injury of hepatic tissues simultaneously.

Effect of pulsed electric field treatment on shelf life and nutritional value of apple juice.

The aim of this study was to assess shelf life and nutritional value of apple juice, including the content of bioactive compounds, after pulsed electric field (PEF) treatment, taking into account different number of cycles: 4, 6, 8 (total 200, 300, and 400 pulses, respectively). Determination of vitamin C and polyphenols concentration, antioxidant activity as well as microbiological analysis were conducted immediately after PEF process and after 24, 48 and 72 h of storage. The results showed that PEF did not affect the content of bioactive compounds. PEF-treated juice did not show changes in the amount of vitamin C and total polyphenols during the storage for 72 h under refrigeration. PEF treatment was effective method for inactivation of a wide range of most common food spoilage microorganisms. PEF process can be used as an effective method of food preservation, allowing prolongation of shelf life and protection of nutritional value. This brings new opportunities for obtaining safe, healthy and nutritious food.

Effect of emergent non-thermal extraction technologies on bioactive individual compounds profile from different plant materials.

Extraction is the first step for isolation and purification of interesting bioactive compounds, by mixing of the plant material with an adequate solvent. Those bioactive compounds are, usually, secondary metabolites, such as phenolic acids and flavonoids which are present in closed insoluble structures, making its extraction a challenge. There are many different traditional extraction methods, such as Soxhlet, heat reflux, and maceration. Nevertheless, due to several disadvantages, they are being replaced by new methods, using emergent technologies, such as high hydrostatic pressure, ultrasounds, pulsed electric fields, and supercritical fluids. The use of novel technologies allows enhancing mass transfer rates, increasing cell permeability as well as increasing secondary metabolite diffusion, leading to higher extraction yields, fewer impurities on the final extract, extractions at room temperature with thermo-sensitive structures preservation, use of different non-organic solvents, low energy consumption, short operation time, and have no significant or lower effect on the structure of bioactive compounds. This paper aims to review the effect of the main emergent extraction technologies (high hydrostatic pressure, pulsed electric fields, ultrasounds, and supercritical fluid assisted) on the individual profile of bioactive compounds from plant material.

From the basic science of biological effects of ultrashort electrical pulses to medical therapies.

This article is based on my presentation at the D'Arsonval Ceremony at the Joint Annual Meeting of the Bioelectromagnetics Society and the European BioElectromagnetics Association in Hangzhou, China, in June of 2017. It describes the pathway from the first studies on the effects of intense, nanosecond pulses on biological cells to the development of medical therapies based on these effects. The motivation for the initial studies of the effects of high voltage, nanosecond pulses on mammalian cells was based on a simple electrical circuit model, which predicted that such pulses allow us to affect not just the plasma membrane but also the subcellular structures. The first experimental study that confirmed this hypothesis was published in 2001 in the Bioelectromagnetics journal. It was followed by a large number of publications that showed that such ultrashort pulses affect cell functions, such as programmed cell death, and, at lower intensity, calcium mobilization from intracellular structures. These basic studies were leading to novel cancer treatments, treatments of cardiac arrhythmia, and advanced wound healing. Further, by reducing the pulse duration into the picosecond range, antenna-based neural stimulation seems to be possible. This manuscript gives an overview of the progress in this field of research in the decade after the initial bioelectric studies with high-voltage, nanosecond pulses, particularly the research performed at the Frank Reidy Research Center for Bioelectrics. It also tells you about my journey and that of my colleagues at the Center for Bioelectrics into and through this fascinating bioelectromagnetics research area. Bioelectromagnetics. 39:257-276, 2018. © 2018 Wiley Periodicals, Inc.

Inactivation kinetics of Escherichia coli in cranberry juice during multistage treatment by electric fields.

The inactivation of Escherichia coli inoculated in cranberry juice by processing with radio frequency electric fields was studied. E. coli ATCC 35218 was chosen among three non-pathogenic strains based on its ability to survive in low pH cranberry juice. Studies were conducted by measuring the survival population when changing the electric field strength between 2.2 and 13.2 kV cm-1, number of treatment stages from 1 to 6 and flow rates between 13 and 25 L h-1 at moderate temperatures of 20, 30 and 40 °C. A minimum inactivation of 5-log reduction, as requested by the Food and Drug Administration (FDA), can be achieved by increasing the number of treatment stages, temperature or both. At 40 °C and 6 treatment stages, 6.57 ±â€¯0.02 log CFU ml-1 reduction in the initial population of E. coli (ATCC 35218) was obtained. At a constant electric field, increasing the temperature produced higher microbial inactivation, consuming lower radio frequency energy input, than increasing the number of treatment stages. Furthermore, a primary model that accounts for the combined effect of time and electric field is proposed. The model represented the sigmoidal curve composed of shoulder, log-linear and tailing sections as observed when changing electric fields. A secondary model that accounts for the effect of temperature and flow rate on the primary model constants is also proposed. The combined primary and secondary models were found to fit the data well with a high coefficient of determination (R2 = 0.965). The proposed model can be extended to kinetic models for pulsed electric fields.

Effect of high-hydrostatic pressure and moderate-intensity pulsed electric field on plum.

Moderate intensity pulse electric fields were applied in plum with the aim to increase bioactive compounds content of the fruit, while high-hydrostatic pressure was applied to preserve the purées. High-hydrostatic pressure treatment was compared with an equivalent thermal treatment. The addition of ascorbic acid during purée manufacture was also evaluated. The main objective of this study was to assess the effects on microorganisms, polyphenoloxidase, color and bioactive compounds of high-hydrostatic pressure, or thermal-processed plum purées made of moderate intensity pulse electric field-treated or no-moderate intensity pulse electric field-treated plums, after processing during storage. The application of moderate intensity pulse electric field to plums slightly increased the levels of anthocyanins and the antioxidant activity of purées. The application of Hydrostatic-high pressure (HHP) increased the levels of bioactive compounds in purées, while the thermal treatment preserved better the color during storage. The addition of ascorbic acid during the manufacture of plum purée was an important factor for the final quality of purées. The color and the bioactive compounds content were better preserved in purées with ascorbic acid. The no inactivation of polyphenoloxidase enzyme with treatments applied in this study affected the stability purées. Probably more intense treatments conditions (high-hydrostatic pressure and thermal treatment) would be necessary to reach better quality and shelf life during storage.

S. Typhimurium virulence changes caused by exposure to different non-thermal preservation treatments using C. elegans.

The aims of this research study were: (i) to postulate Caenorhabditis elegans (C. elegans) as a useful organism to describe infection by Salmonella enterica serovar Typhimurium (S. Typhimurium), and (ii) to evaluate changes in virulence of S. Typhimurium when subjected repetitively to different antimicrobial treatments. Specifically, cauliflower by-product infusion, High Hydrostatic Pressure (HHP), and Pulsed Electric Fields (PEF). This study was carried out by feeding C. elegans with different microbial populations: E. coli OP50 (optimal conditions), untreated S. Typhimurium, S. Typhimurium treated once and three times with cauliflower by-product infusion, S. Typhimurium treated once and four times with HHP and S. Typhimurium treated once and four times with PEF. Bayesian survival analysis was applied to estimate C. elegans lifespan when fed with the different microbial populations considered. Results showed that C. elegans is a useful organism to describe infection by S. Typhimurium because its lifespan was reduced when it was infected. In addition, the application of antimicrobial treatments repetitively generated different responses: when cauliflower by-product infusion and PEF treatment were applied repetitively the virulence of S. Typhimurium was lower than when the treatment was applied once. In contrast, when HHP treatment was applied repetitively, the virulence of S. Typhimurium was higher than when it was applied once. Nevertheless, in all the populations analyzed treated S. Typhimurium had lower virulence than untreated S. Typhimurium.

Innovative "Green" and Novel Strategies for the Extraction of Bioactive Added Value Compounds from Citrus Wastes-A Review.

Citrus is a major processed crop that results in large quantities of wastes and by-products rich in various bioactive compounds such as pectins, water soluble and insoluble antioxidants and essential oils. While some of those wastes are currently valorised by various technologies (yet most are discarded or used for feed), effective, non-toxic and profitable extraction strategies could further significantly promote the valorisation and provide both increased profits and high quality bioactives. The present review will describe and summarize the latest works concerning novel and greener methods for valorisation of citrus by-products. The outcomes and effectiveness of those technologies such as microwaves, ultrasound, pulsed electric fields and high pressure is compared both to conventional valorisation technologies and between the novel technologies themselves in order to highlight the advantages and potential scalability of these so-called "enabling technologies". In many cases the reported novel technologies can enable a valorisation extraction process that is "greener" compared to the conventional technique due to a lower energy consumption and reduced utilization of toxic solvents.

Technological characteristics and selected bioactive compounds of Opuntia dillenii cactus fruit juice following the impact of pulsed electric field pre-treatment.

Selected technological characteristics and bioactive compounds of juice pressed directly from the mash of whole Opuntia dillenii cactus fruits have been investigated. The impact of pulsed electric fields (PEF) for a non-thermal disintegration on the important juice characteristics has been evaluated in comparison to microwave heating and use of pectinases. Results showed that the cactus juice exhibited desirable technological characteristics. Besides, it also contained a high amount of phenolic compounds being the major contributors to the overall antioxidant activity of juice. HPLC-DAD/ESI-MS(n) measurements in the fruits' peel and pulp showed that isorhamnetin 3-O-rutinoside was determined as the single flavonol found only in the fruit's peel. Treating fruit mash with a moderate electric field strength increased juice yield and improved juice characteristics. Promisingly, the highest release of isorhamnetin 3-O-rutinoside from fruit's peel into juice was maximally achieved by PEF.

Effect of pulsed electric fields assisted acetylation on morphological, structural and functional characteristics of potato starch.

Pulsed electric fields (PEF)-assisted acetylation of potato starch with different degree of substitution (DS) was prepared and effects of PEF strength, reaction time, starch concentration on DS were studied by response surface methodology. Results showed DS was increased from 0.054 (reaction time of 15 min) to 0.130 (reaction time of 60 min) as PEF strength increased from 3 to 5 kV/cm. External morphology revealed that acetylated starch with higher DS was aggravated more bulges and asperities. Fourier-transformed infrared spectroscopy confirmed the introduction of acetyl group through a band at 1730 cm(-1). The optimum sample (DS =0 .13) had lower retrogradation (39.1%), breakdown (155 BU) and setback value (149BU), while pasting temperature (62.2 °C) was slightly higher than non-PEF-assisted samples. These results demonstrated PEF treatment can be a potential and beneficial method for acetylation and achieve higher DS with shorter reaction time.

Pulsed electric field improves the bioprotective capacity of purées for different coloured carrot cultivars against H2O2-induced oxidative damage.

This research aimed to study the effect of pulsed electric field (PEF) processing on the bioprotective capacity of carrot purée for White Belgian, Yellow Solar, Nantes, Nutri Red and Purple Haze cultivars against H2O2-induced oxidative damage. The bioprotective capacity was determined using cell viability, membrane integrity and nitric oxide (NO) production in a human Caco-2 cell culture assay. Total carotenoids, total anthocyanins, total vitamin C and total phenolics were also evaluated. Compared to the untreated purée, Purple Haze and Nutri Red processed at 303 kJ/kg completely increased Caco-2 cells resistance towards oxidative damage by recovering the cell viability and inhibiting NO production. For cultivar with low carotenoid levels, i.e. Yellow Solar, the application of 0.8 kV/cm resulted in a higher total carotenoid content in the purée than its untreated counterpart, leading to an improved bioprotective effect. This study clearly shows that PEF could add value to carrots by maximising bioprotective effects.

Bioaccumulation of the Selected Metal Ions in Saccharomyces cerevisiae Cells Under Treatment of the Culture with Pulsed Electric Field (PEF).

The obtained results demonstrated an influence of PEF on increase in accumulation of various ions in S. cerevisiae cells. Optimization of particular PEF parameters and ions concentrations in the medium caused twofold increase in accumulation of magnesium and zinc ions and 3.5-fold higher accumulation of calcium ions in the cells. In the case of ion couple, accumulation of magnesium and zinc was, respectively, 1.5-fold and twofold higher in comparison to the control cultures. Yeast cells biomass enriched with Mg(2+), Zn(2+), Ca(2+) as well as Mg(2+) and Zn(2+) (simultaneously) may be an alternative for pharmacological supplementation applied in deficiency of these cations.