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In this study, we investigated the combined effect of 222 nm krypton-chlorine excilamp (EX) and 307 nm ultraviolet-B (UVB) light on the inactivation of Salmonella Typhimurium and Listeria monocytogenes on sliced cheese. The data confirmed that simultaneous exposure to EX and UVB irradiation for 80 s reduced S. Typhimurium and L. monocytogenes population by 3.50 and 3.20 log CFU/g, respectively, on sliced cheese. The synergistic cell count reductions in S. Typhimurium and L. monocytogenes in the combined treatment group were 0.88 and 0.59 log units, respectively. The inactivation mechanism underlying the EX and UVB combination treatment was evaluated using fluorescent staining. The combination of EX and UVB light induced the inactivation of reactive oxygen species (ROS) defense enzymes (superoxide dismutase) and synergistic ROS generation, resulting in synergistic lipid peroxidation and destruction of the cell membrane. There were no significant (P > 0.05) differences in the color, texture, or sensory attributes of sliced cheese between the combination treatment and control groups. These results demonstrate that combined treatment with EX and UVB light is a potential alternative strategy for inactivating foodborne pathogens in dairy products without affecting their quality.
Assuntos
Queijo , Cloro , Listeria monocytogenes , Espécies Reativas de Oxigênio , Salmonella typhimurium , Raios Ultravioleta , Queijo/microbiologia , Queijo/análise , Listeria monocytogenes/efeitos da radiação , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/efeitos dos fármacos , Salmonella typhimurium/efeitos da radiação , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Cloro/farmacologia , Irradiação de Alimentos/métodos , Microbiologia de Alimentos , Viabilidade Microbiana/efeitos da radiação , Contagem de Colônia MicrobianaRESUMO
The total melanin synthesis in the skin depends on various melanogenic factors, including the number of viable melanocytes, the level of melanogenic enzymes per cell, and the reaction rate of the enzymes. The purpose of this study is to examine the effects of L-cysteine (L-Cys), L-ascorbic acid (L-AA), and their derivatives on the tyrosinase (TYR) activity and autoxidation of L-3,4-dihydroxyphenylalanine (L-DOPA) in vitro and the viability and melanin synthesis of B16/F10 cells under different conditions. L-Cysteinamide (C-NH2), glutathione (GSH), L-Cys, L-AA, and N-acetyl L-cysteine (NAC) inhibited the catalytic activity of TYR in vitro. L-AA, C-NH2, L-ascorbic acid 2-O-glucoside (AAG), and 3-O-ethyl L-ascorbic acid (EAA) inhibited the autoxidation of L-DOPA in vitro. L-DOPA exhibited cytotoxicity at 0.1 mM and higher concentrations, whereas L-tyrosine (L-Tyr) did not affect cell viability up to 3 mM. L-AA, magnesium L-ascorbyl 2-phosphate (MAP), and L-Cys attenuated the cell death induced by L-DOPA. C-NH2 decreased the intracellular melanin level at the basal state, whereas L-AA, MAP, and AAG conversely increased it. C-NH2 reduced the number of darkly pigmented cells via in situ L-DOPA staining, whereas L-AA, MAP, GSH, and AAG increased it. C-NH2 decreased the intracellular melanin level at the alpha-melanocyte-stimulating hormone (α-MSH)-stimulated state, while NAC and GSH increased it. L-AA and C-NH2 decreased the intracellular melanin level at the L-Tyr-stimulated state, but NAC and GSH increased it. L-Ascorbyl tetraisopalmitate (ATI) showed no or minor effects in most experiments. This study suggests that L-AA can either promote or inhibit the different melanogenic factors, and C-NH2 can inhibit the multiple melanogenic factors consistently. This study highlights the multifaceted properties of L-Cys, L-AA, and their derivatives that can direct their therapeutic applications in hyperpigmentation, hypopigmentation, or both disorders.
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Salicylic acid (SA) in different concentration were used to assess its individual effect as well as combined impact with 3% calcium chloride (CaCl2) on Swat No. 8 variety of peach stored at refrigerated temperature (6 ± 2 °C) for 21 days. Interestingly, the results revealed that applying 2 mmol L-1 SA with 3% CaCl2 maintained maximum nutritional value and least decay percent (44.1%) as compared to other treatments during storage. Moreover, this combination also exhibited significant weight loss and chilling injury at 6 ± 2 °C whereas increased levels of total phenolic, flavonoids, tannin, alkaloid, ascorbic acid, and antioxidant activity were observed as compared to other treatments. The combination treatment (2 mmol L-1 SA with 3% CaCl2) maintained other sensory attributes of peach fruit during refrigerated storage, therefore, its commercial use was recommended to store peach fruit for up to three weeks without any significant nutritional and physical loss.
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Metal chelators are used for various industrial and medical purposes based on their physicochemical properties and biological activities. In biological systems, copper ions bind to certain enzymes as cofactors to confer catalytic activity or bind to specific proteins for safe storage and transport. However, unbound free copper ions can catalyze the production of reactive oxygen species (ROS), causing oxidative stress and cell death. The present study aims to identify amino acids with copper chelation activities that might mitigate oxidative stress and toxicity in skin cells exposed to copper ions. A total of 20 free amino acids and 20 amidated amino acids were compared for their copper chelation activities in vitro and the cytoprotective effects in cultured HaCaT keratinocytes exposed to CuSO4. Among the free amino acids, cysteine showed the highest copper chelation activity, followed by histidine and glutamic acid. Among the amidated amino acids, cysteinamide showed the highest copper chelation activity, followed by histidinamide and aspartic acid. CuSO4 (0.4-1.0 mM) caused cell death in a concentration-dependent manner. Among the free and amidated amino acids (1.0 mM), only histidine and histidinamide prevented the HaCaT cell death induced by CuSO4 (1.0 mM). Cysteine and cysteinamide had no cytoprotective effects despite their potent copper-chelating activities. EDTA and GHK-Cu, which were used as reference compounds, had no cytoprotective effects either. Histidine and histidinamide suppressed the CuSO4-induced ROS production, glutathione oxidation, lipid peroxidation, and protein carbonylation in HaCaT cells, whereas cysteine and cysteinamide had no such effects. Bovine serum albumin (BSA) showed copper-chelating activity at 0.5-1.0 mM (34-68 mg mL-1). Histidine, histidinamide, and BSA at 0.5-1.0 mM enhanced the viability of cells exposed to CuCl2 or CuSO4 (0.5 mM or 1.0 mM) whereas cysteine and cysteinamide had no such effects. The results of this study suggest that histidine and histidinamide have more advantageous properties than cysteine and cysteinamide in terms of alleviating copper ion-induced toxic effects in the skin.
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Contaminated food contact surface is one of the most important transmission routes for foodborne pathogens. Stainless steel is one such food-contact surface that is widely used in food-processing environments. The present study aimed to evaluate the synergistic antimicrobial efficacy of a combination of tap water-based neutral electrolyzed water (TNEW) and lactic acid (LA) against the foodborne pathogens Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on stainless steel. The results revealed that simultaneous treatment with TNEW (ACC of 4.60 mg/L) and 0.1% LA (TNEW-LA) for 5 min resulted in 4.99-, 4.34-, and >5.4- log CFU/cm2 reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes on stainless steel, respectively. Of these, 4.00-, 3.57-, and >4.76-log CFU/cm2 reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively were exclusively attributed to the synergistic action of the combined treatments after factoring out the reductions due to individual treatments. Furthermore, five mechanistic investigations revealed that the key mechanisms underlying the synergistic antibacterial effect of TNEW-LA were reactive oxygen species (ROS) production, cell membrane damage resulting from membrane lipid oxidation, DNA damage, and inactivation of intracellular enzymes. Overall, our findings suggest that the TNEW-LA combination treatment could be effectively used in the sanitization of food processing environments, especially the food contact surfaces, to control major pathogens and enhance food safety.
Assuntos
Escherichia coli O157 , Listeria monocytogenes , Salmonella typhimurium , Aço Inoxidável , Microbiologia de Alimentos , Contagem de Colônia Microbiana , Ácido Láctico/farmacologia , Água/farmacologiaRESUMO
Human pathogens can develop biofilm structures on different artificial substrates common in the food industry. In this study, we investigated the inactivation efficacy of low-energy X-ray irradiation on Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes biofilms on food contact surfaces, including polyvinyl chloride (PVC), stainless steel with finish 2B (STS 2B), and Teflon. The numbers of viable cells in biofilms on all test coupons were significantly (p < 0.05) reduced as the X-ray dose increased. Interestingly, different biofilm inactivation levels were observed relative to various material surfaces. Teflon showed the lowest D5d (dose required for a 5-log reduction in cell count) values among three groups of coupons, whereas PVC exhibited higher D5d values than the other two coupons. The mechanism of the X-ray antibiofilm effect was identified through the measurement of extracellular polymeric substances (EPS) in biofilms. X-ray irradiation could remove exopolysaccharides, which are major component of EPS and the removal rate increased with increasing irradiation dose. The analyses also confirmed that the disintegration of EPS was strongly related to the trends of biofilm inactivation on different coupon surfaces. This study is the first to demonstrate that X-ray irradiation effectively inactivates major foodborne pathogen biofilms on various food contact surfaces and to evaluate its antibiofilm mechanisms to enhance safety in the food processing industries.
Assuntos
Microbiologia de Alimentos , Listeria monocytogenes , Biofilmes , Contagem de Colônia Microbiana , Humanos , Politetrafluoretileno/farmacologia , Cloreto de Polivinila/farmacologia , Aço Inoxidável , Raios XRESUMO
Air pollution causes oxidative stress that leads to inflammatory diseases and premature aging of the skin. The purpose of this study was to examine the antioxidant effect of Korean propolis on oxidative stress in human epidermal HaCaT keratinocytes exposed to particulate matter with a diameter of less than 10 µm (PM10). The total ethanol extract of propolis was solvent-fractionated with water and methylene chloride to divide into a hydrophilic fraction and a lipophilic fraction. The lipophilic fraction of propolis was slightly more cytotoxic, and the hydrophilic fraction was much less cytotoxic than the total extract. The hydrophilic fraction did not affect the viability of cells exposed to PM10, but the total propolis extract and the lipophilic fraction aggravated the toxicity of PM10. The total extract and hydrophilic fraction inhibited PM10-induced ROS production and lipid peroxidation in a concentration-dependent manner, whereas the lipophilic fraction did not show such effects. High-performance liquid chromatography with photodiode array detection (HPLC-DAD) analysis showed that the hydrophilic fraction contained phenylpropanoids, such as caffeic acid, p-coumaric acid, and ferulic acid, whereas the lipophilic faction contained caffeic acid phenethyl ester (CAPE). The former three compounds inhibited PM10-induced ROS production, lipid peroxidation, and/or glutathione oxidation, and ferulic acid was the most effective among them, but CAPE exhibited cytotoxicity and aggravated the toxicity of PM10. This study suggests that Korean propolis, when properly purified, has the potential to be used as a cosmetic material that helps to alleviate the skin toxicity of air pollutants.
RESUMO
This study aimed to evaluate the effect of different growth temperatures on the resistance of Listeria monocytogenes and Yersinia enterocolitica to low-energy X-ray irradiation and elucidate the mechanisms of resistance variability. The X-ray treatment at a dose of 1.0 kGy resulted in 4.00-, 4.87-, 3.98-, and 2.27-log reductions in cell counts of L. monocytogenes cultured at 37, 25, 15, and 4 °C, respectively. Cell counts of Y. enterocolitica, cultured at 37, 25, 15, and 4 °C, in phosphate-buffered saline decreased by 3.96, 4.98, 3.79, and 3.25 log CFU/mL, respectively, after X-ray irradiation at 0.4 kGy. In addition, the increased resistance to X-rays at low temperatures (4 °C) was induced by different mechanisms in the two pathogens. The results reveal that the key mechanisms for the change in resistance of L. monocytogenes and Y. enterocolitica to X-ray irradiation under different growth temperatures are efflux pump malfunction and DNA damage, respectively. These results suggest that the stress resistance status of L. monocytogenes and Y. enterocolitica cultured at different growth temperatures (37, 25, 15, and 4 °C) should be considered for application in low-dose X-ray irradiation in the food industry.
Assuntos
Listeria monocytogenes , Yersinia enterocolitica , Temperatura , Raios XRESUMO
As an emerging electrolyzed water (EW) technology, tap water-based neutral electrolyzed water (TNEW) is an attractive alternative to other types of conventional EW for sterilization of food contact surfaces. In this study, we sought to identify strategies for improving TNEW inactivation efficiencies of major foodborne pathogenic bacteria. We investigated the synergistic antimicrobial effect of TNEW and ultraviolet-A light (UVA) combination treatment against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on stainless steel. The data confirmed that simultaneous TNEW and UVA treatment for 60 min reduced E. coli O157:H7, S. Typhimurium, and L. monocytogenes population by 2.15, 1.55, and 2.65 log CFU/cm2, respectively. The synergistic cell count reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes in the combination treatment group were 1.17, 0.59, and 1.62 log units, respectively. Additionally, the mechanisms of the synergistic bactericidal effects of TNEW and UVA were identified through several approaches. Mechanistic investigations suggested that the synergistic effect was associated with intracellular reactive oxygen species generation, bacterial cell membrane damage, and inactivation of dehydrogenase. These findings demonstrate that treatment with TNEW and UVA light can enhance the microbiological safety of food contact surfaces during food processing.
Assuntos
Escherichia coli O157 , Aço Inoxidável , Contagem de Colônia Microbiana , Microbiologia de Alimentos , Viabilidade Microbiana , Raios Ultravioleta , ÁguaRESUMO
Airborne particulate matter with a size of 10 µm or less (PM10) can cause oxidative damages and inflammatory reactions in the skin. This study was conducted to discover natural products that are potentially useful in protecting the skin from PM10. Among the hot water extracts of a total of 23 medicinal plants, Siegesbeckiae Herba extract (SHE), which showed the strongest protective effect against PM10 cytotoxicity, was selected, and its mechanism of action and active constituents were explored. SHE ameliorated PM10-induced cell death, lactate dehydrogenase (LDH) release, lipid peroxidation, and reactive oxygen species (ROS) production in HaCaT cells. SHE decreased the expression of KEAP1, a negative regulator of NRF2, and increased the expression of NRF2 target genes, such as HMOX1 and NQO1. SHE selectively induced the enzymes involved in the synthesis of GSH (GCL-c and GCL-m), the regeneration of GSH (GSR and G6PDH), and GSH conjugation of xenobiotics (GSTκ1), rather than the enzymes that directly scavenge ROS (SOD1, CAT, and GPX1). SHE increased the cellular content of GSH and mitigated the oxidation of GSH to GSSG caused by PM10 exposure. Of the solvent fractions of SHE, the n-butyl alcohol (BA) fraction ameliorated cell death in both the absence and presence of PM10. The BA fraction contained a high amount of chlorogenic acid. Chlorogenic acid reduced PM10-induced cell death, LDH release, and ROS production. This study suggests that SHE protects cells from PM10 toxicity by increasing the cellular antioxidant capacity and that chlorogenic acid may be an active phytochemical of SHE.
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The purpose of this study is to identify amino acid derivatives with potent anti-eumelanogenic activity. First, we compared the effects of twenty different amidated amino acids on tyrosinase (TYR)-mediated dopachrome formation in vitro and melanin content in dark-pigmented human melanoma MNT-1 cells. The results showed that only L-cysteinamide inhibited TYR-mediated dopachrome formation in vitro and reduced the melanin content of cells. Next, the antimelanogenic effect of L-cysteinamide was compared to those of other thiol compounds (L-cysteine, N-acetyl L-cysteine, glutathione, L-cysteine ethyl ester, N-acetyl L-cysteinamide, and cysteamine) and positive controls with known antimelanogenic effects (kojic acid and ß-arbutin). The results showed the unique properties of L-cysteinamide, which effectively reduces melanin content without causing cytotoxicity. L-Cysteinamide did not affect the mRNA and protein levels of TYR, tyrosinase-related protein 1, and dopachrome tautomerase in MNT-1 cells. L-Cysteinamide exhibited similar properties in normal human epidermal melanocytes (HEMs). Experiments using mushroom TYR suggest that L-cysteinamide at certain concentrations can inhibit eumelanin synthesis through a dual mechanism by inhibiting TYR-catalyzed dopaquinone synthesis and by diverting the synthesized dopaquinone to the formation of DOPA-cysteinamide conjugates rather than dopachrome. Finally, L-cysteinamide was shown to increase pheomelanin content while decreasing eumelanin and total melanin contents in MNT-1 cells. This study suggests that L-cysteinamide has an optimal structure that can effectively and safely inhibit eumelanin synthesis in MNT-1 cells and HEMs, and will be useful in controlling skin hyperpigmentation.
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This study aimed to investigate the effect of different growth temperatures on the resistance of Escherichia coli O157:H7 and Salmonella Typhimurium to low-energy X-ray irradiation. Irradiation of contaminated phosphate-buffered saline with 0.6 kGy X-ray decreased the counts of E. coli O157:H7 cultured at 37 °C to below the detection limit (<1.0 colony-forming unit (CFU)/mL) and those of E. coli O157:H7 cultured at 25 and 15 °C by 4.82 and 4.45 log CFU/mL, respectively. The viable counts of S. Typhimurium cultured at 37, 25, and 15 °C in phosphate-buffered saline decreased by 3.56, 3.08, and 2.75 log CFU/mL, respectively, after irradiation with 0.6 kGy X-ray. Irradiation of contaminated lettuce with 0.4 kGy decreased the counts of E. coli O157:H7 cultured at 37, 25, and 15 °C by 3.97, 3.45, and 3.10 log CFU/cm2, respectively, and those of S. Typhimurium by 4.41, 3.84, and 3.40 log CFU/cm2, respectively. Growth temperature influenced pathogen resistance to X-ray irradiation by modulating cellular membrane and DNA integrity, intracellular enzyme activity, and efflux pump function. The results of this study suggest that the stress resistance status of pathogenic bacteria cultured at different growth temperatures should be considered for the application of X-ray irradiation for fresh produce sterilization.
Assuntos
Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/efeitos da radiação , Lactuca/microbiologia , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/efeitos da radiação , Contagem de Colônia Microbiana , Contaminação de Alimentos/prevenção & controle , Irradiação de Alimentos , Folhas de Planta/microbiologia , Temperatura , Raios XRESUMO
The aim of this study was to evaluate the synergistic antimicrobial effect of xenon light (XL) and citric acid (CA) combination against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on spinach leaves and determine the effect of XL-CA combination on quality of spinach leaves. The XL-CA combined treatment for 8 min synergistically decreased the cell counts of E. coli O157:H7 and S. Typhimurium by 5.25 and 5.05 log CFU/cm2, respectively, and additively decreased the L. monocytogenes cells by 5.02 log unit on spinach. The mechanisms underlying synergistic lethal effect of the XL-CA combination were investigated. Qualitative and quantitative analyses revealed that the bacterial cell membrane damage was strongly associated with the synergistic antimicrobial effect of the XL-CA combination. Additionally, treatment with XL-CA combination for 8 min did not affect the quality attributes (color, total phenol contents, and texture) of spinach leaves. These results suggest that the XL-CA combination treatment can be effectively used to control major pathogens on fresh produce.
Assuntos
Microbiologia de Alimentos , Spinacia oleracea , Ácido Cítrico/farmacologia , Contagem de Colônia Microbiana , Folhas de Planta , XenônioRESUMO
The objectives of this study were to evaluate the bactericidal effects of X-ray irradiation and gallic acid (GA) against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on lettuce leaves and in phosphate-buffered saline (PBS). Inoculated PBS and lettuce were exposed to X-rays (0.05, 0.1, and 0.15; 0.1, 0.2, and 0.3 kGy, respectively), and GA was applied to lettuce leaves as a solution and in PBS at concentrations of 0.5% (w/v). Combined treatment with 0.3 kGy and 0.5% GA reduced E. coli O157:H7, S. Typhimurium, and L. monocytogenes cell counts 5.41, 2.57, and 1.36 log CFU/cm2 on lettuce, respectively. Combined treatment with 0.15 kGy X-ray and 0.5% GA reduced counts for the same species by 6.54, 4.24, and 1.51 log CFU/mL in PBS. The combined treatments exerted a synergistic antibacterial effect against E. coli O157:H7 on lettuce, but not against S. Typhimurium or L. monocytogenes. In PBS, the synergistic effect was confirmed in both E. coli O157:H7 and S. Typhimurium cells. Mechanistic investigations indicated that the synergistic antibacterial effect was associated with intracellular reactive oxygen species (ROS) generation and bacterial cell membrane damage. Additionally, the X-ray and GA combination treatment did not adversely affect the color, total phenol content, and texture of lettuce. These findings demonstrate that treatment with X-ray radiation and GA can enhance the microbiological safety of fresh produce.
Assuntos
Antibacterianos/farmacologia , Conservação de Alimentos/métodos , Ácido Gálico/farmacologia , Lactuca/microbiologia , Contagem de Colônia Microbiana , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/efeitos da radiação , Irradiação de Alimentos/métodos , Conservação de Alimentos/instrumentação , Conservantes de Alimentos/farmacologia , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/efeitos da radiação , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/efeitos da radiação , Raios XRESUMO
The aim of this study was to identify novel antimelanogenic drugs from an epigenetic screening library containing various modulators targeting DNA methyltransferases, histone deacetylases, and other related enzymes/proteins. Of 141 drugs tested, K8 (4-((hydroxyamino)carbonyl)-N-(2-hydroxyethyl)-N-phenyl-benzeneacetamide; HPOB) was found to effectively inhibit the α-melanocyte-stimulating hormone (α-MSH)-induced melanin synthesis in B16-F10 murine melanoma cells without accompanying cytotoxicity. Additional experiments showed that K8 did not significantly reduce the mRNA and protein level of tyrosinase (TYR) or microphthalmia-associated transcription factor (MITF) in cells, but it potently inhibited the catalytic activity TYR in vitro (IC50, 1.1-1.5 µM) as compared to ß-arbutin (IC50, 500-700 µM) or kojic acid (IC50, 63 µM). K8 showed copper chelating activity similar to kojic acid. Therefore, these data suggest that K8 inhibits cellular melanin synthesis not by downregulation of TYR protein expression through an epigenetic mechanism, but by direct inhibition of TYR catalytic activity through copper chelation. Metal chelating activity of K8 is not surprising because it is known to inhibit histone deacetylase (HDAC) 6 through zinc chelation. This study identified K8 as a potent inhibitor of cellular melanin synthesis, which may be useful for the treatment of hyperpigmentation disorders.
Assuntos
Antineoplásicos/farmacologia , Benzenoacetamidas/química , Epigênese Genética , Melaninas/metabolismo , Melanoma Experimental/tratamento farmacológico , Monofenol Mono-Oxigenase/antagonistas & inibidores , Preparações Farmacêuticas/administração & dosagem , Animais , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Melanoma Experimental/metabolismo , Melanoma Experimental/patologia , Camundongos , Fator de Transcrição Associado à Microftalmia/genética , Fator de Transcrição Associado à Microftalmia/metabolismo , Preparações Farmacêuticas/isolamento & purificação , Células Tumorais CultivadasRESUMO
In this study, we investigated the antimicrobial activity of the X-ray irradiation and citric acid (CA) combination against Escherichia coli O157:H7 and Listeria monocytogenes on the surface of spinach leaves and elucidated the mechanisms underlying their synergistic interaction. Upon treatment with 0.3 kGy X-ray irradiation and 1% CA combination, the cell counts of E. coli O157:H7 and L. monocytogenes reduced by 4.23 and 3.69 log CFU/mL on spinach leaves, respectively. The synergistic reduction in the cell counts of E. coli O157:H7 and L. monocytogenes by the combination treatment was 0.95 and 1.14 log units, respectively. The X-ray and CA combination exerts its antimicrobial effect by damaging the bacterial cell membrane and enhancing the generation of intracellular reactive oxygen species in the pathogens. The enhanced bactericidal effect of the combination treatment may not be due to the loss of intracellular enzyme activity. We also evaluated the effect of the combination treatment on the quality attributes of spinach leaves. The combination treatment did not result in adverse changes in color and texture of spinach leaves. These results demonstrate the potential of citric acid and X-ray irradiation combination for decontaminating foodborne pathogens on fresh produce.
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Ácido Cítrico/farmacologia , Desinfetantes/farmacologia , Irradiação de Alimentos/métodos , Spinacia oleracea/microbiologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/efeitos da radiação , Contagem de Colônia Microbiana , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/metabolismo , Escherichia coli O157/efeitos da radiação , Microbiologia de Alimentos , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/metabolismo , Listeria monocytogenes/efeitos da radiação , Folhas de Planta/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Raios XRESUMO
We evaluated the bactericidal efficacy of the simultaneous application of ultraviolet-A (UV-A) irradiation and fumaric acid (FA) against Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes in apple juice and as well as investigated the effects of this treatment on product quality. Further, we elucidated the mechanisms underlying their synergistic bactericidal action. Simultaneous UV-A light irradiation and 0.1% FA treatment for 30 min resulted in 6.65-, 6.27-, and 6.49-log CFU/ml reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, which involved 3.15, 2.21, and 3.43 log CFU reductions, respectively, and these were attributed to the synergistic action of the combined treatments. Mechanistic investigations suggested that the combined UVA-FA treatment resulted in significantly greater bacterial cell membrane damage and intracellular reactive oxygen species (ROS) generation. UVA-FA treatment for 30 min did not cause significant changes to the color, nonenzymatic browning index, pH, and total phenolic content of apple juice. These results suggest that combined UVA-FA treatment can be effectively used to control foodborne pathogens in apple juice without affecting its quality.
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Antibacterianos/farmacologia , Conservação de Alimentos/métodos , Sucos de Frutas e Vegetais/microbiologia , Fumaratos/farmacologia , Malus/microbiologia , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/metabolismo , Escherichia coli O157/efeitos da radiação , Conservação de Alimentos/instrumentação , Sucos de Frutas e Vegetais/análise , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/metabolismo , Listeria monocytogenes/efeitos da radiação , Viabilidade Microbiana/efeitos dos fármacos , Viabilidade Microbiana/efeitos da radiação , Espécies Reativas de Oxigênio/metabolismo , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/metabolismo , Salmonella typhimurium/efeitos da radiação , Raios UltravioletaRESUMO
The objective of this study was to evaluate the efficacy of simultaneous UV-A and UV-B irradiation (UV-A+B) for inactivating Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in both phosphate-buffered saline (PBS) and apple juice. A cocktail of the three pathogens was inoculated into PBS and apple juice, and then the suspensions were irradiated with UV lamps of 356 nm (UV-A) and 307 nm (UV-B). Significant (P < 0.05) log reductions of the three pathogens in PBS and apple juice were observed after a maximum dose of UV-B alone or the UV-A+B treatment, but few reductions were observed upon UV-A treatment alone. At all irradiation times, antagonistic effects were observed for the application of UV-A+B against in E. coli O157:H7, Salmonella Typhimurium, and L. monocytogenes in PBS and apple juice. The degree of antagonistic effect in apple juice was greater than that in PBS. The results of this study suggest that the combined treatment of commercial UV-A and UV-B lamps would be impractical for disinfecting juice products.
Assuntos
Escherichia coli O157 , Microbiologia de Alimentos , Listeria monocytogenes , Viabilidade Microbiana , Salmonella typhimurium , Raios Ultravioleta , Contagem de Colônia Microbiana , Escherichia coli O157/efeitos da radiação , Microbiologia de Alimentos/métodos , Sucos de Frutas e Vegetais/microbiologia , Listeria monocytogenes/efeitos da radiação , Malus , Viabilidade Microbiana/efeitos da radiação , Salmonella typhimurium/efeitos da radiaçãoRESUMO
The purpose of this study was to evaluate the synergistic bactericidal efficacy of combining ultrasound (US) and fumaric acid (FA) treatment against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes in apple juice and to identify the synergistic bactericidal mechanisms. Additionally, the effect of combination treatment on juice quality was determined by measuring the changes in color, pH, non-enzymatic browning index, and total phenolic content. A mixed cocktail of the three pathogens was inoculated into apple juice, followed by treatment with US (40â¯kHz) alone, FA (0.05, 0.1, and 0.15%) alone, and a combination of US and FA for 1, 2, 3, 4, and 5â¯min. Combined US and 0.15% FA treatment for 5â¯min achieved 5.67, 6.35, and 3.47 log reductions in E. coli O157:H7, S. Typhimurium, and L. monocytogenes, respectively, with the 1.55, 2.37, and 0.57 log CFU reductions attributed to the synergistic effect. Although the pH value slightly decreased as FA increased, there were no significant (Pâ¯>â¯0.05) differences in color values, browning indices, and phenolic content between untreated and treated samples. To identify the mechanism of this synergistic bactericidal action, membrane integrity, malfunctions in the membrane efflux pump, and intracellular enzyme activity were measured. The analyses confirmed that damage to the cell envelope (membrane integrity and efflux pump) was strongly related to the synergistic microbial inactivation. These results suggest that simultaneous application of US treatment and FA is a novel method for ensuring the microbial safety of apple juice.
Assuntos
Antibacterianos/farmacologia , Bactérias/efeitos da radiação , Sucos de Frutas e Vegetais/microbiologia , Fumaratos/farmacologia , Malus/microbiologia , Viabilidade Microbiana/efeitos da radiação , Ondas Ultrassônicas , Bactérias/patogenicidade , Contagem de Colônia Microbiana , Escherichia coli O157/efeitos dos fármacos , Escherichia coli O157/efeitos da radiação , Microbiologia de Alimentos/métodos , Conservação de Alimentos/métodos , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/efeitos da radiação , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/efeitos da radiaçãoRESUMO
Atmospheric particulate matter (PM) is an important cause of skin damage, and an increasing number of studies have been conducted to discover safe, natural materials that can alleviate the oxidative stress and inflammation caused by PM. It has been previously shown that the extract of Ecklonia cava Kjellman, a perennial brown macroalga, can alleviate oxidative stress in epidermal keratinocytes exposed to PM less than 10 microns in diameter (PM10). The present study was undertaken to further examine the anti-inflammatory effects of E. cava extract and its major polyphenolic constituent, dieckol. HaCaT keratinocytes were exposed to PM10 in the presence or absence of E. cava extract or dieckol and analyzed for their viability, prostaglandin E2 (PGE2) release, and gene expression of cyclooxygenase (COX)-1, COX-2, microsomal prostaglandin E2 synthase (mPGES)-1, mPGES-2, and cytosolic prostaglandin E2 synthase (cPGES). PM10 treatment decreased cell viability and increased the production of PGE2, and these changes were partially abrogated by E. cava extract. E. cava extract also attenuated the expression of COX-1, COX-2, and mPGES-2 stimulated by PM10. Dieckol attenuated PGE2 production and the gene expression of COX-1, COX-2, and mPGES-1 stimulated by PM10. This study demonstrates that E. cava extract and dieckol alleviate airborne PM10-induced PGE2 production in keratinocytes through the inhibition of gene expression of COX-1, COX-2, mPGES-1, and/or mPGES-2. Thus, E. cava extract and dieckol are potentially useful natural cosmetic ingredients for counteracting the pro-inflammatory effects of airborne PM.