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BACKGROUND: Tomato (Solanum lycopersicum) has a relatively short shelf life as a result of rapid ripening, limiting its transportability and marketability. Recently, gamma irradiation has emerged as a viable method for delaying tomato fruit ripening. Although few studies have shown that gamma irradiation delays the ripening of tomatoes, the underlying mechanism remains unknown. Therefore, the present study aimed to examine the effects of gamma irradiation on tomato fruit ripening and the underlying mechanisms using transcriptomics. RESULTS: Following gamma irradiation, the total microbial count, weight loss, and decay rate of tomatoes significantly reduced during storage. Furthermore, the redness (a*), color change (∆E), and lycopene content of gamma-irradiated tomatoes decreased in a dose-dependent manner during storage. Moreover, gamma irradiation significantly upregulated the expression levels of genes associated with DNA, chloroplast, and oxidative damage repairs, whereas those of ethylene and auxin signaling-, ripening-, and cell wall metabolism-related, as well as carotenoid genes, were downregulated. CONCLUSION: Gamma irradiation effectively delayed ripening by downregulating the expression of ripening-related genes and inhibiting microbial growth, which prevented decay and prolonged the shelf life of tomatoes. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
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Solanum lycopersicum , Solanum lycopersicum/genética , Etilenos/metabolismo , Carotenoides/análisis , Licopeno/análisis , Pared Celular/metabolismo , Frutas/química , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
IMPORTANCE: Weaning is a crucial step in piglet management to improve pork production. During the weaning phase, disruption of epithelial barrier function and intestinal inflammation can lead to decreased absorption of nutrients and diarrhea. Therefore, maintaining a healthy intestine, epithelial barrier function, and gut microbiota composition in this crucial phase is strategic for optimal weaning in pigs. We isolated a lysate of Lactococcus petauri GB97 (LPL97) from healthy porcine feces and evaluated its anti-inflammatory activities, barrier integrity, and gut microbial changes in LPS-induced murine macrophages and DSS-induced colitis mice. We found that LPL97 regulated the immune response by downregulating the TLR4/NF-κB/MAPK signaling pathway both in vitro and in vivo. Furthermore, LPL97 alleviated the disruption of intestinal epithelial integrity and gut microbiota dysbiosis in colitis mice. This study indicates that LPL97 has the potential to be developed as an alternative feed additive to antibiotics for the swine industry.
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Colitis , Microbioma Gastrointestinal , Lactococcus , Porcinos , Animales , Ratones , Funcion de la Barrera Intestinal , Inflamación , Colitis/inducido químicamente , Heces , Modelos Animales de EnfermedadRESUMEN
This study aimed to evaluate the efficacy of Lactiplantibacillus argentoratensis AGMB00912 (LA) in reducing Salmonella Typhimurium infection in weaned piglets. The investigation focused on the influence of LA on the gut microbiota composition, growth performance, and Salmonella fecal shedding. The results indicated that LA supplementation significantly improved average daily gain and reduced the prevalence and severity of diarrhea. Fecal analysis revealed reduced Salmonella shedding in the LA-supplemented group. Furthermore, LA notably altered the composition of the gut microbiota, increasing the levels of beneficial Bacillus and decreasing those of harmful Proteobacteria and Spirochaetes. Histopathological examination showed less intestinal damage in LA-treated piglets than in the controls. The study also observed that LA affected metabolic functions related to carbohydrate, amino acid, and fatty acid metabolism, thereby enhancing gut health and resilience against infection. Short-chain fatty acid concentrations in the feces were higher in the LA group, suggesting improved gut microbial activity. LA supplementation enriched the population of beneficial bacteria, including Streptococcus, Clostridium, and Bifidobacterium, while reducing the number of harmful bacteria, such as Escherichia and Campylobacter. These findings indicate the potential of LA as a probiotic alternative for swine nutrition, offering protective effects to the gut microbiota against Salmonella infection.
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Heces , Microbioma Gastrointestinal , Probióticos , Destete , Animales , Microbioma Gastrointestinal/efectos de los fármacos , Porcinos , Proyectos Piloto , Probióticos/administración & dosificación , Heces/microbiología , Salmonelosis Animal/microbiología , Enfermedades de los Porcinos/microbiología , Enfermedades de los Porcinos/prevención & control , Lactobacillaceae , Salmonella typhimurium/efectos de los fármacosRESUMEN
Polyphenolic compounds have been found to possess a wide range of physiological activities that may contribute to their beneficial effects against inflammation-related diseases; however, the molecular mechanisms underlying this anti-inflammatory activity are not completely characterized, and many features remain to be elucidated. In this study, we investigated the molecular basis for the down-regulation of toll-like receptor 4 (TLR4) signal transduction by procyanidin dimer B2 (Pro B2) in macrophages. Pro B2 markedly elevated the expression of the interleukin (IL)-1 receptor-associated kinase (IRAK)-M protein, a negative regulator of TLR signaling. Lipopolysaccharide (LPS)-induced expression of cell surface molecules (CD80, CD86, and MHC class I/II) and production of pro-inflammatory cytokines (tumor necrosis factor-α, IL-1ß, IL-6, and IL-12p70) were inhibited by Pro B2, and this action was prevented by IRAK-M silencing. In addition, Pro B2-treated macrophages inhibited LPS-induced activation of mitogen-activated protein kinases such as extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase and the translocation of nuclear factor κB and p65 through IRAK-M. We also found that Pro B2-treated macrophages inactivated naïve T cells by inhibiting LPS-induced interferon-γ and IL-2 secretion through IRAK-M. These novel findings provide new insights into the understanding of negative regulatory mechanisms of the TLR4 signaling pathway and the immune-pharmacological role of Pro B2 in the immune response against the development and progression of many chronic diseases.
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Biflavonoides/farmacología , Catequina/farmacología , Quinasas Asociadas a Receptores de Interleucina-1/biosíntesis , Macrófagos/metabolismo , Proantocianidinas/farmacología , Receptor Toll-Like 4/metabolismo , Animales , Línea Celular , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/fisiología , Macrófagos/efectos de los fármacos , RatonesRESUMEN
We have investigated the use of BMSC (bone marrow stromal cell) as a feeder cell for improving culture efficiency of ESC (embryonic stem cell). B6CBAF1 blastocysts or ESC stored after their establishment were seeded on to a feeder layer of either SCA-1+/CD45-/CD11b- BMSC or MEF (mouse embryonic fibroblast). Feeder cell activity in promoting ESC establishment from the blastocysts and in supporting ESC maintenance did not differ significantly between BMSC and MEF feeders. However, the highest efficiency of colony formation after culturing of inner cell mass cells of blastocysts was observed with the BMSC line that secreted the largest amount of LIF (leukaemia inhibitory factor). Exogenous LIF was essential for the ESC establishment on BMSC feeder, but not for ESC maintenance. Neither change in stem cell-specific gene expression nor increase in stem cell aneuploidy was detected after the use of BMSC feeder. We conclude that BMSC can be utilized as the feeder of ESC, which improves culture efficiency.
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Masa Celular Interna del Blastocisto/citología , Células de la Médula Ósea/metabolismo , Células Madre Embrionarias/fisiología , Fibroblastos/metabolismo , Células del Estroma/metabolismo , Animales , Antígenos de Diferenciación/genética , Antígenos de Diferenciación/metabolismo , Diferenciación Celular , Proliferación Celular , Técnicas de Cocultivo , Medios de Cultivo Condicionados/química , Células Madre Embrionarias/trasplante , Células Nutrientes , Fémur/citología , Expresión Génica , Cariotipo , Factor Inhibidor de Leucemia/metabolismo , Ratones , Ratones Endogámicos ICR , Ratones Endogámicos NOD , Ratones SCID , Tibia/citologíaRESUMEN
Primary follicles retrieved from B6CBAF1 prepubertal mice were cultured in a stepwise manner in an alpha-minimum essential medium-based medium to generate viable embryos and embryonic stem cell (ESC)-like cells. A significant increase in follicle growth and oocyte maturation accompanied by increased secretion of 17beta-estradiol and progesterone was achieved by exposing primary follicles to 100 or 200 mIU of follicle-stimulating hormone (FSH) during culture. More oocytes developed into blastocysts following in vitro fertilization (IVF) or parthenogenetic activation after culture with 200 mIU of FSH during the entire culture period than with 100 mIU. Eleven ESC-like cell lines, consisting of four heterozygotic and seven homozygotic phenotypes, were established from 25 trials of primary follicle culture combined with IVF or parthenogenetic activation. In conclusion, primary follicles can potentially yield developmentally competent oocytes, which produce viable embryos and ESC-like cell lines following in vitro manipulation. We suggest a method to utilize immature follicles, which are most abundant in ovaries, to improve reproductive efficiency and for use in regenerative medicine.
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Blastocisto/citología , Ectogénesis , Células Madre Embrionarias/citología , Técnicas de Maduración In Vitro de los Oocitos , Oogénesis , Folículo Ovárico/citología , Animales , Biomarcadores/metabolismo , Blastocisto/metabolismo , Línea Celular , Supervivencia Celular , Cruzamientos Genéticos , Células Madre Embrionarias/metabolismo , Estradiol/metabolismo , Femenino , Fertilización In Vitro , Hormona Folículo Estimulante/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Folículo Ovárico/metabolismo , Partenogénesis , Progesterona/metabolismo , Técnicas de Cultivo de Tejidos , Supervivencia TisularRESUMEN
Polycyclic aromatic hydrocarbons (PAHs) are suspect human lung carcinogens and can be metabolically activated to remote quinones, for example, benzo[a]pyrene-1,6-dione (B[a]P-1,6-dione) and B[a]P-3,6-dione by the action of either P450 monooxygenase or peroxidases, and to non-K region o-quinones, for example B[a]P-7,8-dione, by the action of aldo keto reductases (AKRs). B[a]P-7,8-dione also structurally resembles 4-hydroxyequilenin o-quinone. These three classes of quinones can redox cycle, generate reactive oxygen species (ROS), and produce the mutagenic lesion 8-oxo-dGuo and may contribute to PAH- and estrogen-induced carcinogenesis. We compared the ability of a complete panel of human recombinant AKRs to catalyze the reduction of PAH o-quinones in the phenanthrene, chrysene, pyrene, and anthracene series. The specific activities for NADPH-dependent quinone reduction were often 100-1000 times greater than the ability of the same AKR isoform to oxidize the cognate PAH-trans-dihydrodiol. However, the AKR with the highest quinone reductase activity for a particular PAH o-quinone was not always identical to the AKR isoform with the highest dihydrodiol dehydrogenase activity for the respective PAH-trans-dihydrodiol. Discrete AKRs also catalyzed the reduction of B[a]P-1,6-dione, B[a]P-3,6-dione, and 4-hydroxyequilenin o-quinone. Concurrent measurements of oxygen consumption, superoxide anion, and hydrogen peroxide formation established that ROS were produced as a result of the redox cycling. When compared with human recombinant NAD(P)H:quinone oxidoreductase (NQO1) and carbonyl reductases (CBR1 and CBR3), NQO1 was a superior catalyst of these reactions followed by AKRs and last CBR1 and CBR3. In A549 cells, two-electron reduction of PAH o-quinones causes intracellular ROS formation. ROS formation was unaffected by the addition of dicumarol, suggesting that NQO1 is not responsible for the two-electron reduction observed and does not offer protection against ROS formation from PAH o-quinones.
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Oxidorreductasas de Alcohol/metabolismo , Equilenina/análogos & derivados , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Hidrocarburos Policíclicos Aromáticos/metabolismo , Quinonas/metabolismo , Oxidorreductasas de Alcohol/genética , Aldehído Reductasa , Aldo-Ceto Reductasas , Benzopirenos/química , Benzopirenos/toxicidad , Biocatálisis , Línea Celular Tumoral , Equilenina/química , Equilenina/metabolismo , Equilenina/toxicidad , Humanos , Isomerismo , NAD(P)H Deshidrogenasa (Quinona)/genética , Oxidación-Reducción/efectos de los fármacos , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/toxicidad , Quinonas/química , Quinonas/toxicidad , Especies Reactivas de Oxígeno/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidad por SustratoRESUMEN
Polycyclic aromatic hydrocarbons (PAHs) are tobacco carcinogens implicated in the causation of human lung cancer. Metabolic activation is a key prerequisite for PAHs to cause their deleterious effects. Using human lung adenocarcinoma (A549) cells, we provide evidence for the metabolic activation of (+/-)-trans-7,8dihydroxy-7,8-dihydrobenzo[a]pyrene (B[a]P-7,8-trans-dihydrodiol) by aldo-keto reductases (AKRs) to yield benzo[a]pyrene-7,8-dione (B[a]P-7,8-dione), a redox-active o-quinone. We show that B[a]P-7,8-trans-dihydrodiol (AKR substrate) and B[a]P-7,8-dione (AKR product) lead to the production of intracellular reactive oxygen species (ROS) (measured as an increase in dichlorofluorescin diacetate fluores-cence) and that similar changes were not observed with the regioisomer (+/-)-trans-4,5-dihydroxy-4,5-dihydrobenzo[a]pyrene or the diol-epoxide, (+/-)-anti-7,8-dihydroxy-9alpha,10beta-epoxy-7,8,9,10-tetrahydro-B[a]P. B[a]P-7,8-trans-dihydrodiol and B[a]P-7,8-dione also caused a decrease in glutathione levels and an increase in NADP(+)/NADPH ratios, with a concomitant increase in single-strand breaks (as measured by the comet assay) and 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dGuo). The specificity of the comet assay was validated by coupling it to human 8-oxo-guanine glycosylase (hOGG1), which excises 8-oxo-Gua to yield single-strand breaks. The levels of 8-oxo-dGuo observed were confirmed by an immunoaffinity purification stable isotope dilution ([(15)N(5)]-8-oxo-dGuo) liquid chromatography-electrospray ionization/multiple reaction monitoring/mass spectrometry (LC-ESI/MRM/MS) assay. B[a]P-7,8-trans-dihydrodiol produced DNA strand breaks in the hOGG1-coupled comet assay as well as 8-oxo-dGuo (as measured by LC-ESI/MRM/MS) and was enhanced by a catechol O-methyl transferase (COMT) inhibitor, suggesting that COMT protects against o-quinone-mediated redox cycling. We conclude that activation of PAH-trans-dihydrodiols by AKRs in lung cells leads to ROS-mediated genotoxicity and contributes to lung carcinogenesis.
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Oxidorreductasas de Alcohol/metabolismo , Dihidroxidihidrobenzopirenos/metabolismo , Pulmón/enzimología , 8-Hidroxi-2'-Desoxicoguanosina , Aldehído Reductasa , Aldo-Ceto Reductasas , Benzopirenos/farmacología , Biotransformación/efectos de los fármacos , Inhibidores de Catecol O-Metiltransferasa , Línea Celular Tumoral , Ensayo Cometa , Roturas del ADN de Doble Cadena/efectos de los fármacos , ADN Glicosilasas/metabolismo , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Dihidroxidihidrobenzopirenos/farmacología , Inhibidores Enzimáticos/farmacología , Fluoresceínas/metabolismo , Fluorescencia , Humanos , Isoenzimas/metabolismo , Pulmón/patología , Oxidación-Reducción/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Reproducibilidad de los Resultados , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Polycyclic aromatic hydrocarbon (PAH) o-quinones produced by aldo-keto reductases are ligands for the aryl hydrocarbon receptor (AhR) (Burczynski, M. E., and Penning, T. M. (2000) Cancer Res. 60, 908-915). They induce oxidative DNA lesions (reactive oxygen species-mediated DNA strand breaks and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dGuo) formation) in human lung cells. We tested whether the AhR enhances PAH o-quinone-mediated oxidative DNA damage by translocating these ligands to the nucleus. Using the single cell gel electrophoresis (comet) assay to detect DNA strand breaks in murine hepatoma Hepa1c1c7 cells and its AhR- and aryl hydrocarbon receptor nuclear translocator-deficient variants, benzo[a]pyrene-7,8-dione (B[a]P-7,8-dione) produced fewer DNA strand breaks in AhR-deficient cells compared with aryl hydrocarbon receptor nuclear translocator-deficient and wild type Hepa1c1c7 cells. Decreased DNA strand breaks were also observed in human bronchoalveolar H358 cells in which the AhR was silenced by siRNA. The antioxidant alpha-tocopherol and the iron chelator/antioxidant desferal decreased the formation of B[a]P-7,8-dione-mediated DNA strand breaks indicating that they were reactive oxygen species-dependent. By coupling the comet assay to 8-oxoguanine glycosylase (hOGG1), which excises 8-oxo-Gua, strand breaks dependent upon this lesion were measured. hOGG1 treatment produced more DNA single strand breaks in B[a]P-7,8-dione-treated Hepa cells and H358 cells than in its absence. The levels of hOGG1-dependent DNA strand breaks mediated by B[a]P-7,8-dione were lower in AhR-deficient Hepa and AhR knockdown H358 cells. The AhR antagonist alpha-naphthoflavone also attenuated B[a]P-7,8-dione-mediated DNA strand breaks. The decrease in 8-oxo-dGuo levels in AhR-deficient Hepa cells and AhR knockdown H358 cells was validated by immunoaffinity capture stable isotope dilution ([(15)N(5)]8-oxo-dGuo) liquid chromatography-electrospray ionization/multiple reaction monitoring/mass spectrometry. We conclude that the AhR shuttles PAH o-quinone genotoxins to the nucleus and enhances oxidative DNA damage.
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Benzopirenos/farmacología , Roturas del ADN/efectos de los fármacos , Desoxiguanosina/análogos & derivados , Especies Reactivas de Oxígeno/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Antioxidantes/farmacología , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Línea Celular , Línea Celular Tumoral , ADN Glicosilasas/genética , ADN Glicosilasas/metabolismo , Deferoxamina/farmacología , Desoxiguanosina/genética , Desoxiguanosina/metabolismo , Humanos , Ratones , Receptores de Hidrocarburo de Aril/genética , Sideróforos/farmacología , alfa-Tocoferol/farmacologíaRESUMEN
Mistletoe offers health-promoting effects; however, it has toxicity, requiring careful application. Viscothionin is a polypeptide of mistletoe that while contributing to toxicity also demonstrates anti-cancer and anti-diabetic activities. The aim of this study was to evaluate whether gamma irradiation or heating treatment could selectively reduce viscothionin-mediated cytotoxicity. Gamma irradiation effectively inhibited viscothionin-induced cytotoxicity to RIN5mF cells, but heating treatment did not affect its cytotoxicity. Both heating and gamma irradiation further increased the insulinotropic activity of viscothionin, whereas the effect of gamma irradiation was dose-dependent and diminished above 20 kGy. Structural analysis showed that gamma irradiation significantly altered the ordered structure of viscothionin, unlike heating treatment, resulting in a change of its molecular properties, which could be linked to the observed changes in the cytotoxicity and insulinotropic activity of the polypeptide. These results suggest gamma irradiation as an alternative method for minimizing viscothionin toxicity without interfering with anti-diabetic effect.
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This study evaluated the applicability of a rapid analytical method using a headspace solid-phase microextraction gas chromatography/mass spectrometry (HS-SPME-GC/MS) technique to identify gamma-irradiated soybeans (0.1-5 kGy). From the partial least squares discriminant analysis used to discriminate between non-irradiated and irradiated soybean samples, 1,7-hexadecadiene was selected as the identifying marker. Response surface methodology experiments were used to determine the optimal HS-SPME extraction conditions including a carboxen/polydimethylsiloxane fiber with an extraction temperature of 98 °C and an extraction time of 55 min. 1,7-Hexdecadiene was detected in all samples irradiated at ≥ 0.1 kGy under the optimized HS-SPME-GC/MS conditions, and the unique presence of the marker in a gamma-irradiated sample was verified by comparing the results from heat, steam, microwave, sonication, and ultraviolet treatments. The comparisons of the identification properties for various conventional methods validated several advances in HS-SPME-GC/MS analysis in terms of rapid analysis, high sensitivity, and absence of solvent.
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Cromatografía de Gases y Espectrometría de Masas/métodos , Glycine max/química , Glycine max/efectos de la radiación , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Microextracción en Fase Sólida/métodos , Rayos gammaRESUMEN
This study evaluated the effects of X-ray irradiation (0-1 kGy) on quality parameters of Korean strawberries during storage at 15 °C for 9 d. As compared to control, all irradiated samples exhibited dose-dependent decreases in microbial counts regardless of storage period. Irradiation significantly (p < 0.05) reduced weight loss and decay incidence of fruits during storage. Fruit firmness decreased immediately after irradiation, but no significant changes occurred after 3 d. Neither irradiation nor storage period significantly affected total soluble solids, pH, or titratable acidity. All treatments delayed color changes and pelargonidin accumulation during storage. The radical scavenging activities and total phenolic, ellagic acid, and catechin contents increased gradually during storage. Furthermore, irradiated fruits showed improved sensory characteristics throughout storage. Thus, X-ray irradiation (≤1 kGy) was confirmed as a effective phytosanitary treatment for strawberries to delay decay and negative physicochemical changes and extend shelf life with acceptable sensory attributes.
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Analysis of cellular 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dGuo) as a biomarker of oxidative DNA damage has been fraught with numerous methodological problems. This is primarily due to artifactual oxidation of dGuo that occurs during DNA isolation and hydrolysis. Therefore, it has become necessary to rely on using the comet assay, which is not necessarily specific for 8-oxo-dGuo. A highly specific and sensitive method based on immunoaffinity purification and stable isotope dilution liquid chromatography (LC)-multiple reaction monitoring (MRM)/mass spectrometry (MS) that avoids artifact formation has now been developed. Cellular DNA was isolated using cold DNAzol (a proprietary product that contains guanidine thiocyanate) instead of chaotropic- or phenol-based methodology. Chelex-treated buffers were used to prevent Fenton chemistry-mediated generation of reactive oxygen species (ROS) and artifactual oxidation of DNA bases. Deferoxamine was also added to all buffers in order to complex any residual transition metal ions remaining after Chelex treatment. The LC-MRM/MS method was used to determine that the basal 8-oxo-dGuo level in DNA from human bronchoalveolar H358 cells was 2.2 +/- 0.4 8-oxo-dGuo/10(7) dGuo (mean +/- standard deviation) or 5.5 +/- 1.0 8-oxo-dGuo/10(8) nucleotides. Similar levels were observed in human lung adenocarcinoma A549 cells, mouse hepatoma Hepa-1c1c7 cells, and human HeLa cervical epithelial adenocarcinoma cells. These values are an order of magnitude lower than is typically reported for basal 8-oxo-dGuo levels in DNA as determined by other MS- or chromatography-based assays. H358 cells were treated with increasing concentrations of potassium bromate (KBrO3) as a positive control or with the methylating agent methyl methanesulfonate (MMS) as a negative control. A linear dose-response for 8-oxo-dGuo formation (r(2) = 0.962) was obtained with increasing concentrations of KBrO3 in the range of 0.05 mM to 2.50 mM. In contrast, no 8-oxo-dGuo was observed in H358 cell DNA after treatment with MMS. At low levels of oxidative DNA damage, there was an excellent correlation between a comet assay that measured DNA single strand breaks (SSBs) after treatment with human 8-oxo-guanine glycosylase-1 (hOGG1) when compared with 8-oxo-dGuo in the DNA as measured by the stable isotope dilution LC-MRM/MS method. Availability of the new LC-MRM/MS assay made it possible to show that the benzo[a]pyrene (B[a]P)-derived quinone, B[a]P-7,8-dione, could induce 8-oxo-dGuo formation in H358 cells. This most likely occurred through redox cycling between B[a]P-7,8-dione and B[a]P-7,8-catechol with concomitant generation of DNA damaging ROS. In keeping with this concept, inhibition of catechol-O-methyl transferase (COMT)-mediated detoxification of B[a]P-7,8-catechol with Ro 410961 caused increased 8-oxo-dGuo formation in the H358 cell DNA.
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Cromatografía Líquida de Alta Presión/métodos , Daño del ADN , Desoxiguanosina/análogos & derivados , Estrés Oxidativo , Espectrometría de Masa por Ionización de Electrospray/métodos , 8-Hidroxi-2'-Desoxicoguanosina , Animales , Línea Celular Tumoral , Cromatografía de Afinidad , Ensayo Cometa , Desoxiguanosina/análisis , Desoxiguanosina/inmunología , Desoxiguanosina/aislamiento & purificación , Humanos , RatonesRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Mistletoe (Viscum album), an evergreen parasitic plant, has been widely used as an oriental phytomedicine to treat diabetes mellitus. However, it is unknown which mistletoe constituent exerts the beneficial effect against the disease. In this study, we examined the hypoglycemic activity of mistletoe and investigated whether the polypeptide viscothionin, purified from mistletoe, was responsible for the activity. MATERIALS AND METHODS: Mistletoe extracts were prepared by heating mistletoe powder made of leaves and twigs in water for 3, 6, 9, and 12â¯h. Rat insulinoma RINm5F cells were used to test the cytotoxicity of the extracts and their effects on the secretion of insulin and its precursor, C-peptide. The inhibitory effects of a mistletoe extract on glucose absorption were measured using an α-glucosidase inhibition assay. To determine the component of mistletoe responsible for the observed effects, the mistletoe extract was precipitated with ethanol or hydrolyzed with a protease for further testing. A potential active constituent of mistletoe was isolated by chromatography and molecular weight cut-off fractionation, and its ability to induce insulin secretion was investigated. RESULTS: A 12-h heat-treated mistletoe extract, showing no cytotoxicity, significantly increased the secretion of insulin and C-peptide by RINm5F cells and enhanced the expression of glucose transporter type 4 (GLUT-4), insulin receptor substrate 1 (IRS-1), and protein kinase B (also known as AKT) in differentiated C2C12 cells. The extract also inhibited α-glucosidase activity. After ethanol precipitation, the extract showed much stronger effects on insulin- and C-peptide-secreting activities of cells, whereas the enzyme-hydrolyzed extract was less effective than the original extract, suggesting that the effect was mediated by a proteinaceous constituent of mistletoe. Subsequent analysis showed that viscothionin, a heat-stable 6-kDa polypeptide isolated from mistletoe, increased the level of insulin secretion by more than 20-fold compared to that induced by the extract. CONCLUSIONS: Our study indicates that the hypoglycemic effect of mistletoe is mediated by its insulinotropic action and α-glucosidase inhibitory activity, and the effect is due to viscothionin, one of the major bioactive constituents of mistletoe.
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Secreción de Insulina/efectos de los fármacos , Células Secretoras de Insulina/efectos de los fármacos , Péptidos/farmacología , Viscum album/química , Animales , Péptido C/metabolismo , Línea Celular Tumoral , Glucosa/metabolismo , Hipoglucemiantes/aislamiento & purificación , Hipoglucemiantes/farmacología , Células Secretoras de Insulina/metabolismo , Insulinoma/metabolismo , Ratones , Mioblastos/efectos de los fármacos , Mioblastos/metabolismo , Péptidos/aislamiento & purificación , Extractos Vegetales/química , Extractos Vegetales/farmacología , Ratas , Factores de TiempoRESUMEN
Whitefly pests, including the sweetpotato whitefly, Bemisia tabaci (Gennadius), and the greenhouse whitefly, Trialeurodes vaporariorum (Westwood), are economically important in agriculture. With the annual growth of the domestic fresh fruit export market, various quarantine treatment methods are being used to export strawberries of better quality. The objective of the present study was to evaluate the effects of gamma rays on the development and reproductive sterility of B. tabaci and T. vaporariorum. In both species, the eggs were completely inhibited from hatching at 50 Gy, and the emergence of third-instar nymphs was completely suppressed at 150 Gy. Some adult B. tabaci and T. vaporariorum spawning occurred at 100 and 70 Gy, respectively; however, at these irradiation levels, F1 hatchability was completely inhibited. Dosimetry results showed that the penetrating power of gamma ray in the strawberry-filled box was the lowest at the mid-box position. Therefore, B. tabaci and T. vaporariorum were placed in the middle of the strawberry-filled box and irradiated. A gamma-ray irradiation of 100 Gy suppressed the development and reproduction of eggs and adults in both B. tabaci and T. vaporariorum. Our data suggest that at least 100 Gy should be used for the control of these two species of whitefly for strawberry export.
Asunto(s)
Fragaria , Hemípteros , Animales , NinfaRESUMEN
PAHs (polycyclic aromatic hydrocarbons) are suspect lung cancer carcinogens that must be metabolically converted into DNA-reactive metabolites. P4501A1/P4501B1 plus epoxide hydrolase activate PAH to (+/-)- anti-benzo[ a]pyrene diol epoxide ((+/-)- anti-BPDE), which causes bulky DNA adducts. Alternatively, aldo-keto reductases (AKRs) convert intermediate PAH trans-dihydrodiols to o-quinones, which cause DNA damage by generating reactive oxygen species (ROS). In lung cancer, the types or pattern of mutations in p53 are predominantly G to T transversions. The locations of these mutations form a distinct spectrum characterized by single point mutations in a number of hotspots located in the DNA binding domain. One route to the G to T transversions is via oxidative DNA damage. An RP-HPLC-ECD assay was used to detect the formation of 8-oxo-dGuo in p53 cDNA exposed to representative quinones, BP-7,8-dione, BA-3,4-dione, and DMBA-3,4-dione under redox cycling conditions. Concurrently, a yeast reporter system was used to detect mutations in the same cDNA samples. Nanomolar concentrations of PAH o-quinones generated 8-oxo-dGuo (detected by HPLC-ECD) in a concentration dependent manner that correlated in a linear fashion with mutagenic frequency. By contrast, micromolar concentrations of (+/-)- anti-BPDE generated (+)- trans- anti-BPDE-N (2)-dGuo adducts (detected by stable-isotope dilution LC/MS methodology) in p53 cDNA that correlated in a linear fashion with mutagenic frequency, but no 8-oxo-dGuo was detected. Previous studies found that mutations observed with PAH o-quinones were predominately G to T transversions and those observed with (+/-)- anti-BPDE were predominately G to C transversions. However, mutations at guanine bases observed with either PAH-treatment occurred randomly throughout the DNA-binding domain of p53. Here, we find that when the mutants were screened for dominance, the dominant mutations clustered at or near hotspots primarily at the protein-DNA interface, whereas the recessive mutations are scattered throughout the DNA binding domain without resembling the spectra observed in cancer. These observations, if extended to mammalian cells, suggest that mutagenesis can drive the pattern of mutations but that biological selection for dominant mutations drives the spectrum of mutations observed in p53 in lung cancer.
Asunto(s)
Desoxiguanosina/análogos & derivados , Mutagénesis/efectos de los fármacos , Mutación/genética , Hidrocarburos Policíclicos Aromáticos/química , Quinonas/química , Quinonas/farmacología , Proteína p53 Supresora de Tumor/genética , 8-Hidroxi-2'-Desoxicoguanosina , ADN/genética , Desoxiguanosina/metabolismo , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Modelos Moleculares , Estructura Terciaria de Proteína , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteína p53 Supresora de Tumor/química , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
This study was conducted to evaluate the toxic effects and potency of 2-dodecylcyclobutanone (2-dDCB), a unique compound derived from palmitic acid via irradiation. In a series of assays of bacterial reverse-mutation, in vitro chromosomal aberration, and in vivo micronucleus, negative responses were found by the treatment of 2-dDCB comparing vehicle control, dimethyl sulfoxide or corn oil. In the acute oral toxicity test, all of the mice administrated 2-dDCB survived, and there were no clinical and necropsy signs observed at any doses (0, 300, and 2000â¯mg/kg body weight) during the experimental period of 14 days. These results suggested that 2-dDCB is a relatively non-toxic substance with median lethality dose higher than 2000â¯mg/kg body weight. Moreover, there were no adverse effects noted in rats orally administrated 2-dDCB everyday via gavage for 28 days, even at the highest dose (2.0â¯mg/kg body weight/day) tested, which is 1000-times higher than the human daily intake of 2-dDCB estimated through an extreme exposure scenario. Overall, these results indicate that 2-dDCB is not likely to raise any human health concerns and irradiated foods containing palmitic acid can be recognized as safe for human consumption under the current international regulation systems for food irradiation.
Asunto(s)
Ciclobutanos/toxicidad , Ácido Palmítico/química , Pruebas de Toxicidad , Animales , Aberraciones Cromosómicas/efectos de los fármacos , Ciclobutanos/administración & dosificación , Ciclobutanos/química , Esquema de Medicación , Femenino , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Salmonella typhimurium/efectos de los fármacosRESUMEN
The purpose of this study was to verify the reliability of photostimulated luminescence (PSL) and thermoluminescence (TL) methods for identifying irradiated foods, described in the European standards EN 13751:2002 and EN 1788:2001, respectively, which were established solely through interlaboratory studies on gamma-irradiated food. Red pepper powder samples irradiated with electron-beams (e-beams), gamma rays and high-energy X-rays were used as model foods. Samples irradiated with each radiation type at ⩾4 kGy could be correctly identified by the PSL method, whereas samples irradiated at ⩾0.5 kGy with each radiation type could be correctly recognized by the TL method when e-beams, gamma rays, or high-energy X-rays were used as normalization sources. However, different TL intensities were observed for minerals separated from red pepper powder for different irradiation sources, which was confirmed using pure quartz and K-feldspar minerals. Further interlaboratory studies are required to verify this phenomenon.
Asunto(s)
Capsicum/química , Irradiación de Alimentos , Mediciones Luminiscentes/métodos , Silicatos de Aluminio/química , Compuestos de Potasio/química , Polvos , Cuarzo/química , Rayos XRESUMEN
Since Korean mistletoe (Viscum album) has been used for alleviating metabolic diseases, it may also prevent the impairment of energy, glucose, lipid, and bone metabolisms in an estrogen-deficient animal model. We determined that long-term consumption of Korean mistletoe water extract (KME) can alleviate menopausal symptoms such as hot flush, increased abdominal fat mass, dyslipidemia, hyperglycemia, and decreased bone mineral density in ovariectomized (OVX) rats fed a high-fat diet, and explored the mechanisms of the effects. OVX rats were divided into four groups and fed high-fat diets supplemented with either 0.6% dextrin (control), 0.2% lyophilized KME + 0.4% dextrin (KME-L), or 0.6% lyophilized KME (KME-H). Sham rats were fed with the high-fat diets with 0.6% dextrin as a normal-control without estrogen deficiency. After eight weeks, OVX rats exhibited impaired energy, glucose and lipid metabolism, and decreased uterine and bone masses. KME-L did not alleviate energy dysfunction. However, KME-H lowered serum levels of total-, LDL-cholesterol, and triglycerides and elevated serum HDL-cholesterol levels in OVX rats with dyslipidemia, to similar levels as normal-control rats. Furthermore, KME-H improved HOMA-IR, an indicator of insulin resistance, in OVX rats. Surprisingly, KME-H fed rats had greater lean mass in the abdomen and leg without differences in fat mass but neither dosage of KME altered bone mineral density in the lumbar spine and femur. The increased lean mass was related to greater phosphorylation of mTOR and eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) in the quadriceps muscles. Hepatic triglyceride contents were lowered with KME-H in OVX rats by increasing carnitine palmitoyltransferase-1 (CPT-1) expression and decreasing fatty acid synthase (FAS) and sterol regulatory element-binding protein-1c (SREBP-1c) expression. In conclusion, KME may be useful for preventing some menopausal symptoms such as hot flushes, dyslipidemia, hepatic steatosis, and loss of muscle mass in post-menopausal women.
Asunto(s)
Dislipidemias/prevención & control , Hígado Graso/prevención & control , Sofocos/prevención & control , Atrofia Muscular/prevención & control , Ovariectomía , Extractos Vegetales/uso terapéutico , Viscum album , Animales , Suplementos Dietéticos , Modelos Animales de Enfermedad , Dislipidemias/metabolismo , Estrógenos/deficiencia , Hígado Graso/metabolismo , Femenino , Sofocos/metabolismo , Hiperglucemia/metabolismo , Hiperglucemia/prevención & control , Corea (Geográfico) , Menopausia/metabolismo , Atrofia Muscular/metabolismo , Osteoporosis/metabolismo , Osteoporosis/prevención & control , Extractos Vegetales/administración & dosificación , Ratas , Ratas Sprague-DawleyRESUMEN
The present study investigated the effects of viscothionin, a compound isolated from Korean mistletoe (Viscum album coloratum), on nonalcoholic fatty liver disease (NAFLD) in both in vitro and in vivo models. A connection was discovered between viscothionin and the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, which is involved in lipid metabolism. Viscothionin was shown to significantly attenuate lipid accumulation in HepG2 cells treated with oleic acid, which induces lipid accumulation. Moreover, the phosphorylation of AMPK and acetyl-coenzyme A carboxylase in HepG2 cells was increased by viscothionin treatment. Viscothionin was orally administered to high fat diet-induced obese mice and subsequently histopathological analysis associated with AMPK signaling pathways was evaluated. A significant reduction in the extent of hepatic steatosis was revealed in viscothionin-treated obese mice. Thus, viscothionin mediates its beneficial effects on NAFLD via AMPK signaling pathways, suggesting that it may be a potential target for novel NAFLD treatments.