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1.
Molecules ; 29(7)2024 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-38611882

RESUMO

This study evaluated the impact of pulsed electric fields (PEFs) combined with three-phase partitioning (TPP) extraction methods on the physicochemical properties, functional properties, and structural characterization of the soluble dietary fiber (SDF) derived from peanut shells (PS). The findings of this study indicated that the application of a PEF-TPP treatment leads to a notable improvement in both the extraction yield and purity of SDF. Consequently, the PEF-TPP treatment resulted in the formation of more intricate and permeable structures, a decrease in molecular weight, and an increase in thermal stability compared to SDFs without TPP treatment. An analysis revealed that the PEF-TPP method resulted in an increase in the levels of arabinose and galacturonic acid, leading to enhanced antioxidant capacities. Specifically, the IC50 values were lower in SDFs which underwent PEF-TPP (4.42 for DPPH and 5.07 mg/mL for ABTS) compared to those precipitated with 40% alcohol (5.54 mg/mL for DPPH, 5.56 mg/mL for ABTS) and PEF75 (6.60 mg/mL for DPPH, 7.61 mg/mL for ABTS), respectively. Notably, the SDFs which underwent PEF-TPP demonstrated the highest water- and oil-holding capacity, swelling capacity, emulsifying activity, emulsion stability, glucose adsorption, pancreatic lipase inhibition, cholesterol adsorption, nitric ion adsorption capacity, and the least gelation concentration. Based on the synthesis scores obtained through PCA (0.536 > -0.030 > -0.33), which indicated that SDFs which underwent PEF-TPP exhibited the highest level of quality, the findings indicate that PEF-TPP exhibits potential and promise as a method for preparing SDFs.


Assuntos
Antioxidantes , Arachis , Benzotiazóis , Ácidos Sulfônicos , Adsorção , Fibras na Dieta
2.
Molecules ; 29(10)2024 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-38792161

RESUMO

Olive oil production is characterized by large amounts of waste, and yet is considerably highly valued. Olive pomace can serve as a cheap source of bioactive compounds (BACs) with important antioxidant activity. Novel technologies like Pulsed Electric Fields (PEF) and High Pressure (HP) and microwave (MW) processing are considered green alternatives for the recovery of BACs. Different microwave (150-600 W), PEF (1-5 kV/cm field strength, 100-1500 pulses/15 µs width), and HP (250-650 MPa) conditions, in various product/solvent ratios, methanol concentrations, extraction temperatures, and processing times were investigated. Results indicated that the optimal MW extraction conditions were 300 W at 50 °C for 5 min using 60% v/v methanol with a product/solvent ratio of 1:10 g/mL. Similarly, the mix of 40% v/v methanol with olive pomace, treated at 650 MPa for the time needed for pressure build-up (1 min) were considered as optimal extraction conditions in the case of HP, while for PEF the optimal conditions were 60% v/v methanol with a product/solvent ratio of 1:10 g/mL, treated at 5000 pulses, followed by 1 h extraction under stirring conditions. Therefore, these alternative extraction technologies could assist the conventional practice in minimizing waste production and simultaneously align with the requirements of the circular bioeconomy concept.


Assuntos
Antioxidantes , Eletricidade , Micro-Ondas , Olea , Pressão , Olea/química , Antioxidantes/química , Antioxidantes/isolamento & purificação , Extratos Vegetais/química , Solventes/química , Azeite de Oliva/química , Metanol/química
3.
Molecules ; 29(16)2024 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-39203004

RESUMO

By increasing the permeability of the cell membrane of the treated material, pulsed electric fields (PEF) enhance the internal transport of various chemical substances. Changing the distribution of these components can modify the chemical and thermal properties of the given material. This study aimed to analyze the impact of PEF (1 kV/cm; 1 and 4 kJ/kg) applied to strawberries prior to drying by various methods (convective, infrared-convective, microwave-convective, and vacuum) on the chemical and thermal properties of the obtained dried materials (sugars content, total phenolic content, and antioxidant capacity (ABTS and DPPH assays); thermal properties (TGA and DSC); and molecular composition (FTIR)). PEF could have induced and/or enhanced sucrose inversion because, compared to untreated samples, PEF-pretreated samples were characterized by a lower share of sucrose in the total sugar content but a higher share of glucose and fructose. Reduced exposure to oxygen and decreased drying temperature during vacuum drying led to obtaining dried strawberries with the highest content of antioxidant compounds, which are sensitive to these factors. All PEF-pretreated dried strawberries exhibited a lower glass transition temperature (Tg) than the untreated samples, which confirms the increased mobility of the system after the application of an electric field.


Assuntos
Antioxidantes , Fragaria , Fragaria/química , Antioxidantes/química , Antioxidantes/análise , Eletricidade , Dessecação/métodos , Sacarose/química , Fenóis/análise , Fenóis/química , Espectroscopia de Infravermelho com Transformada de Fourier
4.
Europace ; 25(6)2023 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-37335976

RESUMO

AIMS: Pulsed field ablation (PFA) has emerged as a promising alternative to thermal ablation for treatment of atrial fibrillation (AF). We report performance and safety using the CENTAURI™ System (Galvanize Therapeutics) with three commercial, focal ablation catheters. METHODS AND RESULTS: ECLIPSE AF (NCT04523545) was a prospective, single-arm, multi-centre study evaluating safety and acute and chronic pulmonary vein isolation (PVI) durability using the CENTAURI System in conjunction with the TactiCath SE, StablePoint, and ThermoCool ST ablation catheters. Patients with paroxysmal or persistent AF were treated at two centres. Patients were analysed in five cohorts based upon ablation settings, catheter, and mapping system. Pulsed field ablation was performed in 82 patients (74% male, 42 paroxysmal AF). Pulmonary vein isolation was achieved in 100% of pulmonary veins (322/322) with first-pass isolation in 92.2% (297/322). There were four serious adverse events of interest (three vascular access complications and one lacunar stroke). Eighty patients (98%) underwent invasive remapping. Pulsed field ablation development Cohorts 1 and 2 showed a per-patient isolation rate of 38% and 26% and a per-PV isolation rate of 47% and 53%, respectively. Optimized PFA Cohorts 3-5 showed a per-patient isolation rate of 60%, 73%, and 81% and a per-PV isolation rate of 84%, 90%, and 92%, respectively. CONCLUSION: ECLIPSE AF demonstrated that optimized PFA using the CENTAURI System with three commercial, contact force-sensing, solid-tip focal ablation catheters resulted in transmural lesion formation and high proportion of durable PVI with a favourable safety profile, thus providing a viable treatment option for AF that integrates with contemporary focal ablation workflows.


Assuntos
Fibrilação Atrial , Ablação por Cateter , Veias Pulmonares , Humanos , Masculino , Feminino , Fibrilação Atrial/diagnóstico , Fibrilação Atrial/cirurgia , Fibrilação Atrial/etiologia , Estudos Prospectivos , Adesões Focais , Resultado do Tratamento , Catéteres , Ablação por Cateter/efeitos adversos , Ablação por Cateter/métodos , Veias Pulmonares/cirurgia , Recidiva
5.
Respiration ; 102(11): 948-960, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37906995

RESUMO

Patients living with chronic bronchitis (CB) suffer from physical limitations and poor quality of life. In general, treatment options that directly address the mucus hypersecretion component of CB are quite limited. Chronic airway inflammation and the associated hypersecretion and cough that are pathognomonic for CB generally result from long-term exposure to airway irritants such as tobacco use and other environmental insults. This, in turn, results in an increase in the quantity and change in composition of the airway mucosa as a consequence of altered goblet cells, club cells, and submucosal glands. Pulsed electric fields (PEFs) provide a method for eradicating the cellular constituents of tissue with limited impact on the stromal proteins. Preclinical evidence in porcine airways demonstrated that particular PEF waveforms allowed for salutary remodeling of the epithelial and submucosal airway tissue layers and appeared to foster rapid regeneration and recovery of the tissue. Therefore, a therapeutic opportunity might exist whereby the application of a specific form of PEF may result in a reduction of the cellular secretory constituents of the airway while also reducing airway mucosal inflammation. This review discusses the use of such PEF to address the underlying disease processes in CB including challenges around device design, dosing, and appropriate delivery methods. Further, we outline considerations for the transition to human airways along with a brief examination of the initial work treating CB patients, suggesting that the therapy is well tolerated with limited adverse events.


Assuntos
Bronquite Crônica , Humanos , Animais , Suínos , Bronquite Crônica/terapia , Bronquite Crônica/metabolismo , Qualidade de Vida , Muco/metabolismo , Células Caliciformes/metabolismo , Inflamação/metabolismo , Mucosa/metabolismo
6.
Int J Mol Sci ; 24(21)2023 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-37958601

RESUMO

In recent years, the application of pulsed electric fields with very short durations (nanoseconds) and extremely high amplitudes (MV/m) has been investigated for novel medical purposes. Various electric protocols have been explored for different objectives, including the utilization of fractionated pulse doses to enhance cell electrosensitization to the uptake of different markers or an increase in apoptosis. This study focused on the use of fluorescence imaging to examine molecular calcium fluxes induced by different fractionated protocols of short electric pulses in neuroblastoma (SH-SY5Y) and mesenchymal stem cells (HaMSCs) that were electroporated using nanosecond pulsed electric fields. In our experimental setup, we did not observe cell electrosensitization in terms of an increase in calcium flux following the administration of fractionated doses of nanosecond pulsed electric fields with respect to the non-fractionated dose. However, we observed the targeted activation of calcium-dependent genes (c-FOS, c-JUN, EGR1, NURR-1, ß3-TUBULIN) based on the duration of calcium flux, independent of the instantaneous levels achieved but solely dependent on the final plateau reached. This level of control may have potential applications in various medical and biological treatments that rely on calcium and the delivery of nanosecond pulsed electric fields.


Assuntos
Cálcio , Neuroblastoma , Humanos , Neuroblastoma/terapia , Apoptose , Genes fos , Transdução de Sinais , Cálcio da Dieta
7.
Electromagn Biol Med ; 41(1): 25-34, 2022 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-34541970

RESUMO

The study aimed to evaluate the possibility to perform electrochemotherapy using nanosecond pulsed electric field (nsPEF) and low electric field (LEF) monopolar electrical impulses to alleviate the problems of conventional electroporation. Two types of pulses have been used to treat MCF-7 human breast carcinoma cell line: very low voltage (electric field strength) long trains of short unipolar electric pulses, and low frequencies of extremely intense (40kV/cm), ultra-short (10ns) electric pulses. The electropermeabilization efficiency of the formed endocytotic vesicles was measured using the cloning efficacy test. The cell viability was decreased significantly at a repetition frequency begins from 0.01 Hz by ~35% and reached complete cell loss at 1 Hz of nanosecond pulses for cells treated before with monopolar pulses at 20 V/cm in the presence of BLM with 4 µM concentration. The uptake of non-permeant drugs has been done without plasma membrane permeabilization (classical electroporation), but by endocytosis. Nanosecond electric pulses can disrupt the membrane of endocytotic vesicles and release the cytotoxic drug bleomycin.


Assuntos
Antineoplásicos , Neoplasias da Mama , Eletroquimioterapia , Antineoplásicos/farmacologia , Neoplasias da Mama/terapia , Sobrevivência Celular , Eletroporação , Feminino , Humanos
8.
J Membr Biol ; 254(2): 141-156, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33427940

RESUMO

Nanosecond pulsed electric fields (nsPEFs) induce changes in the plasma membrane (PM), including PM permeabilization (termed nanoporation), allowing free passage of ions into the cell and, in certain cases, cell death. Recent studies from our laboratory show that the composition of the PM is a critical determinant of PM nanoporation. Thus, we hypothesized that the biological response to nsPEF exposure could be influenced by lipid microdomains, including caveolae, which are specialized invaginations of the PM that are enriched in cholesterol and contain aggregates of important cell signaling proteins, such as caveolin-1 (Cav1). Caveolae play a significant role in cellular signal transduction, including control of calcium influx and cell death by interaction of Cav1 with regulatory signaling proteins. Present results show that depletion of Cav1 increased the influx of calcium, while Cav1 overexpression produced the opposite effect. Additionally, Cav1 is known to bind and sequester important cell signaling proteins within caveolae, rendering the binding partners inactive. Imaging of the PM after nsPEF exposure showed localized depletion of PM Cav1 and results of co-immunoprecipitation studies showed dissociation of two critical Cav1 binding partners (transient receptor potential cation channel subfamily C1 (TRPC1) and inositol trisphosphate receptor (IP3R)) after exposure to nsPEFs. Release of TRPC1 and IP3R from Cav1 would activate downstream signaling cascades, including store-operated calcium entry, which could explain the influx in calcium after nsPEF exposure. Results of the current study establish a significant relationship between Cav1 and the activation of cell signaling pathways in response to nsPEFs.


Assuntos
Sinalização do Cálcio , Caveolina 1 , Membrana Celular/fisiologia , Eletricidade , Cálcio , Cavéolas , Caveolina 1/genética , Canais de Cátion TRPC
9.
Crit Rev Food Sci Nutr ; 61(10): 1738-1750, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32406247

RESUMO

The food processing produces a great amount of wastes that are rich in nutrients. Extraction is the first and most important step in recovery and purification of active ingredients from these wastes. The traditional extraction technologies are known to be laborious and time-consuming, require large volumes of organic solvent, have high temperature and energy costs, and obtain relatively low extraction efficiency. In recent 10 years, a novel, efficient and green extraction method, pulsed electric fields (PEFs) continuous extraction, which is emerging non-thermal food-processing technology, has shown great promise in extracting these food wastes. This work gives an overview of development in the use of PEF continuous extraction for obtaining bioactive ingredients from food-processing wastes. The technology is described in detail with respect to the mechanism, equipment, critical parameters. The protocols and applications of the technology in the extraction of food-processing wastes are comprehensively summarized. Finally, the degradation of bioactive ingredients, industrial applications, problem of novel food, consumer acceptance, and future trends of the technology are discussed. The PEF continuous extraction is considered as the ideal technology of high efficiency and low temperature for natural ingredients extraction. The technology possesses many remarkable potential applications in the food-processing industries compared to the conventional extraction methods.


Assuntos
Eletricidade , Manipulação de Alimentos , Tecnologia de Alimentos , Indústria de Processamento de Alimentos
10.
Lasers Surg Med ; 53(10): 1301-1306, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34008877

RESUMO

BACKGROUND AND OBJECTIVES: This study describes the effects of nano-pulse stimulation (NPS) technology on the common verruca with the objectives of demonstrating efficacy and safety. NPS technology applies nanosecond pulses of non-thermal electrical energy to induce highly localized regulated cell death in the cellular structures of the targeted zone with negligible effects on surrounding non-cellular structures. Previous clinical studies applying NPS to common, benign skin lesions have demonstrated safety and efficacy in clearing seborrheic keratoses and sebaceous hyperplasia. STUDY DESIGN/MATERIALS AND METHODS: Sixty-two subjects were enrolled at a total of five sites. One hundred and ninety-five study verrucae up to 10 mm wide were treated with NPS delivered by a console-based handheld applicator (CellFX® System; Pulse Biosciences) and follow-ups occurred every 30 days with the option to retreat at 30, 60, and 90 days. There were 62 untreated controls and 46% of the treated verrucae were recalcitrant. RESULTS: Overall, 75.3% (70/93) of the common verrucae, 72.7% (8/11) of the flat verrucae, and 43.8% (14/32) of the plantar verrucae treated with NPS were completely clear by 60 days following the last treatment and did not recur within the 120-day observation period. The majority (54%) of verrucae cleared with a single NPS procedure. The most common treatment site reactions were erythema (50.5%) and eschar formation (23.4%) on Day 30 and on Day 120 mild erythema was present in 14% of the cases and hyperpigmentation in 18.5%. No serious adverse events were reported. A particle counter was used during 11 NPS procedures on verrucae and no significant plume generation was detected during these procedures. CONCLUSIONS: NPS is a safe and effective procedure for removing non-genital, cutaneous verrucae. Lasers Surg. Med. © 2021 The Authors. Lasers in Surgery and Medicine published by Wiley Periodicals LLC.


Assuntos
Dermatopatias , Verrugas , Humanos , Pele , Verrugas/terapia
11.
Molecules ; 26(15)2021 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-34361654

RESUMO

Growing attention to environmental protection leads food industries to adopt a model of "circular economy" applying safe and sustainable technologies to recover, recycle and valorize by-products. Therefore, by-products become raw material for other industries. Tomato processing industry produces significant amounts of by-products, consisting of skins and seeds. Tomato skin is very rich in lycopene, and from its seeds, high nutritional oil can be extracted. Alternative use of the two fractions not only could cut disposal costs but also allow one to extract bioactive compounds and an oil with a high nutritional value. This review focused on the recent advance in extraction of lycopene, whose beneficial effects on health are widely recognized.


Assuntos
Antioxidantes/isolamento & purificação , Manipulação de Alimentos/métodos , Licopeno/isolamento & purificação , Solanum lycopersicum , Solanum lycopersicum/química , Solanum lycopersicum/metabolismo
12.
Surg Endosc ; 34(2): 580-589, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31011863

RESUMO

BACKGROUND: Irreversible electroporation (IRE) is an emerging tissue ablation technique, which is safe for sites where thermal-basis techniques are not suitable. The aim of this study is to evaluate the safety and efficacy of magnetic anchoring electrode (MAE)-assisted IRE for normal gastric tissue ablation in a rabbit model. METHODS: IRE (500 V, 100 µs, 99 pulses, 1 Hz) of the gastric wall was performed in 24 adult New Zealand rabbits with a novel catheter-mounted MAE with fluoroscopy and a surgical approach. Procedure time, procedure-related bleeding, perforation, and other complications were recorded. Animals were sacrificed at 30 min, 1 day, 3 days, 7 days, 14 days, and 28 days post-IRE. The stomach was removed en bloc, and the diameter of each lesion was measured. Histopathological analyses by Hematoxylin-Eosin (H&E), masson trichrome, alpha-smooth muscle action (α-SMA), and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) were performed. RESULTS: Gastric tissue ablation with MAE-assisted IRE was successfully performed without any interruption. No perforation or bleeding was observed during IRE or throughout the follow-up period. A demarcated hemorrhage was found in the ablated area upon gross examination. H&E staining showed complete cell death with inflammatory infiltration, edema, and hemorrhaging. TUNEL presented diffuse positive cells in the ablated area. The tissue scaffold was well preserved without damage as indicated by Masson trichrome staining. Ulceration was observed starting from 3 days post-IRE. The mucosal layer was gradually recovered and regenerated within 14-28 days. No other complication was observed post-IRE. CONCLUSIONS: MAE-assisted IRE is safe and effective for normal gastric tissue ablation and the gastric wall recovered in 14-28 days post-IRE.


Assuntos
Técnicas de Ablação/instrumentação , Eletrodos , Eletroporação/instrumentação , Mucosa Gástrica/cirurgia , Magnetismo , Animais , Modelos Animais de Doenças , Eletroporação/métodos , Fenômenos Magnéticos , Coelhos , Coloração e Rotulagem , Estômago/cirurgia
13.
Lasers Surg Med ; 52(4): 315-322, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31376199

RESUMO

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.


Assuntos
Estruturas Celulares/efeitos da radiação , Terapia por Estimulação Elétrica/métodos , Morte Celular Regulada/efeitos da radiação , Pele/efeitos da radiação , Adulto , Caspases/metabolismo , Estruturas Celulares/patologia , Relação Dose-Resposta à Radiação , Feminino , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Pele/metabolismo , Pele/patologia
14.
J Cosmet Laser Ther ; 22(4-5): 195-199, 2020 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-33026267

RESUMO

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.


Assuntos
Terapia por Estimulação Elétrica , Dermatopatias/terapia , Pele , Derme/citologia , Epiderme , Face , Folículo Piloso , Humanos
15.
Int J Mol Sci ; 21(17)2020 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-32859066

RESUMO

Granulysin is a protein present in the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, with cytolytic activity against microbes and tumors. Previous work demonstrated the therapeutic effect of the intratumoral injection of recombinant granulysin and of the systemic injection of an immunotoxin between granulysin and the anti-carcinoembryonic antigen single-chain Fv antibody fragment MFE23, which were produced in the yeast Pichia pastoris. In the present work, we developed a second immunotoxin combining granulysin and the anti-Tn antigen single-chain Fv antibody fragment SM3, that could have a broader application in tumor treatment than our previous immunotoxin. In addition, we optimized a method based on electroporation by pulsed electric field (PEF) to extract the remaining intracellular protein from yeast, augmenting the production and purificiation yield. The immunotoxin specifically recognized the Tn antigen on the cell surface. We also compared the thermal stability and the cytotoxic potential of the extracellular and intracellular immunotoxins on Tn-expressing human cell lines, showing that they were similar. Moreover, the bioactivity of both immunotoxins against several Tn+ cell lines was higher than that of granulysin alone.


Assuntos
Antígenos de Diferenciação de Linfócitos T/genética , Antígenos Glicosídicos Associados a Tumores/imunologia , Imunotoxinas/farmacologia , Neoplasias/metabolismo , Saccharomycetales/crescimento & desenvolvimento , Anticorpos de Cadeia Única/genética , Células A549 , Antígenos de Diferenciação de Linfócitos T/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Eletroporação , Humanos , Células Jurkat , Células MCF-7 , Neoplasias/tratamento farmacológico , Engenharia de Proteínas , Proteínas Recombinantes/farmacologia , Saccharomycetales/genética , Anticorpos de Cadeia Única/farmacologia
16.
Int J Mol Sci ; 21(21)2020 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-33171785

RESUMO

Diets rich in phenolic compounds have been associated to reducing the risk of metabolic syndrome and its derived disorders. Fruits are healthy components of the human diet because of their vitamin, mineral, fiber and phenolic profile. However, they have a short shelf-life which is limited by microbiological growth and enzymatic activity. Innovative preservation methods such as high hydrostatic pressure, pulsed electric fields, ultrasound, microwave, cold plasma and ultraviolet light have become popular for the processing of fruits because they can preserve nutritional quality. In this review, the phenolic profile and health potential of 38 Mesoamerican fruits were assessed. Phenolic compounds were classified based on their contribution to the diet as flavonoids, phenolic acids, tannin, lignins and stilbenoids. Due to this composition, fruits showed a wide range of bioactivities which included anti-inflammatory, anti-diabetic, anti-hypertensive and anti-obesity activities, among others. Phenolic content in fruits submitted to innovative food processing technologies depended on parameters such as enzymatic activity, antioxidant capacity, microstructure integrity and cell viability. Innovative technologies could increase phenolic content while assuring microbiological safety by (i) promoting the release of bound phenolic compounds during processing and (ii) inducing the synthesis of phenolic compounds by activation of phenylpropanoid pathway during storage.


Assuntos
Conservação de Alimentos/métodos , Frutas/química , Fenóis/análise , Flavonoides/análise , Manipulação de Alimentos/métodos , Humanos , Hidroxibenzoatos/análise , Hidroxibenzoatos/química , América do Norte , Valor Nutritivo/fisiologia , Fenóis/química
17.
Molecules ; 25(17)2020 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-32867350

RESUMO

Pulsed electric fields (PEF) is a promising technology that allows the selective extraction of high-added value compounds by electroporation. Thus, PEF provides numerous opportunities for the energy efficient isolation of valuable microalgal bioactive substances (i.e., pigments and polyphenols). The efficiency of PEF-assisted extraction combined with aqueous or dimethyl sulfoxide (DMSO) solvents in recovering pigments and polyphenols from microalgae Tetraselmis chuii (T. chuii) and Phaeodactylum tricornutum (P. tricornutum) was evaluated. Two PEF treatments were applied: (1 kV/cm/400 pulses, 3 kV/cm/45 pulses), with a specific energy input of 100 kJ/kg. The total antioxidant capacity (TAC) was positively influenced by the use of DMSO. The highest TAC in the T. chuii culture was achieved at a lower extraction time and electric field than for P. tricornutum. The use of DMSO only improved the polyphenols' extraction for P. tricornutum, whereas the PEF and extraction time were more important for T. chuii. Carotenoids and chlorophyll a were more efficiently extracted using DMSO, while chlorophyll b levels were higher following aqueous extraction for both microalgae. In P. tricornutum, the TAC and pigment extraction efficiency were in general higher at lower extraction times. It can be concluded that PEF may be a promising alternative for the enhancement of the selective extraction of antioxidant bioactive compounds from microalgae.


Assuntos
Antioxidantes/isolamento & purificação , Eletroporação , Microalgas/química , Polifenóis/isolamento & purificação , Clorófitas/química , Diatomáceas/química
18.
J Sci Food Agric ; 100(4): 1653-1661, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31802493

RESUMO

BACKGROUND: The consumption of pesticide-contaminated sour cherries as fruit or juice has become a major health concern, and so the search for alternative processing technologies, such as pulsed electric fields (PEF), ozone (O), and ultrasonication (US) has intensified. The objectives of this experimental study of sour cherry juice were fourfold: (1) to quantify the removal efficiency of new processing technologies (PEF, O, US), and their combinations, for the pesticides chlorpyrifos ethyl, τ-fluvalinate, cyprodinil, pyraclostrobin, and malathion; (2) to detect their impact on physical, bioactive, and sensory properties; (3) to determine their microbial inactivation levels for Escherichia coli O157:H7, Bacillus cereus, Pseudomonas syringae subs. Syringae, and Penicillum expansum; and (4) to jointly optimize multiple responses of physical, quality, and sensory properties, pesticides, and microbial inactivation. RESULTS: Except for all the O treatments, the physical, bioactive and sensory properties of sour cherry juice were not adversely affected by the treatments. The joint optimization suggested PEF1 (24.7 kV cm-1 for 327 µs), PEF2 (24.7 kV cm-1 for 655 µs), PEF2 + O + US, US, and PEF2 + O as the five best treatments. PEF2 + O + US best achieved both pesticide removal and microbial inactivation. CONCLUSION: PEF2 + O + US provided promising reductions in pesticide and microbial loads. © 2019 Society of Chemical Industry.


Assuntos
Manipulação de Alimentos/métodos , Sucos de Frutas e Vegetais/microbiologia , Praguicidas/química , Prunus avium/microbiologia , Bactérias/classificação , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , Contagem de Colônia Microbiana , Contaminação de Alimentos/análise , Manipulação de Alimentos/instrumentação , Sucos de Frutas e Vegetais/análise , Viabilidade Microbiana , Praguicidas/isolamento & purificação , Prunus avium/química , Controle de Qualidade
19.
Compr Rev Food Sci Food Saf ; 19(2): 530-552, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-33325176

RESUMO

Microorganisms (bacteria, yeast, and microalgae) are a promising resource for products of high value such as nutrients, pigments, and enzymes. The majority of these compounds of interest remain inside the cell, thus making it necessary to extract and purify them before use. This review presents the challenges and opportunities in the production of these compounds, the microbial structure and the location of target compounds in the cells, the different procedures proposed for improving extraction of these compounds, and pulsed electric field (PEF)-assisted extraction as alternative to these procedures. PEF is a nonthermal technology that produces a precise action on the cytoplasmic membrane improving the selective release of intracellular compounds while avoiding undesirable consequences of heating on the characteristics and purity of the extracts. PEF pretreatment with low energetic requirements allows for high extraction yields. However, PEF parameters should be tailored to each microbial cell, according to their structure, size, and other factors affecting efficiency. Furthermore, the recent discovery of the triggering effect of enzymatic activity during cell incubation after electroporation opens up the possibility of new implementations of PEF for the recovery of compounds that are bounded or assembled in structures. Similarly, PEF parameters and suspension storage conditions need to be optimized to reach the desired effect. PEF can be applied in continuous flow and is adaptable to industrial equipment, making it feasible for scale-up to large processing capacities.


Assuntos
Bactérias/química , Eletricidade , Microalgas/química , Leveduras/química , Membrana Celular , Eletroporação/métodos
20.
Int J Med Sci ; 16(9): 1271-1282, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31588193

RESUMO

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.


Assuntos
Carcinoma Hepatocelular/terapia , Terapia por Estimulação Elétrica/métodos , Flavonoides/farmacologia , Hepatócitos/efeitos dos fármacos , Neoplasias Hepáticas/terapia , Animais , Carcinoma Hepatocelular/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Flavonoides/efeitos adversos , Hepatócitos/metabolismo , Humanos , Neoplasias Hepáticas/metabolismo , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos Endogâmicos C57BL , Espécies Reativas de Oxigênio/metabolismo
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