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INTRODUCTION: Firefighters, compared to other occupational groups, are exposed more frequently in their working environment not only to physical issues, such as musculoskeletal disease, respiratory disease, and burns but also to mental health issues, such as PTSD and depression. Specifically, Korean firefighters experience significantly higher rates of work-related injuries compared to those in other countries. Recent statistics from the Korea National Fire Agency indicate a steady increase in the number of firefighting work-related injuries. However, there is a shortage of measures in place to address these issues. This study aims to investigate the health needs, overall healthcare usage, and unmet needs of firefighters in Korea. We also aim to investigate, through in-depth interviews, perceptions and hindering factors for integrative medicine approaches to fulfilling unmet needs. METHOD: This study was conducted in accordance with the consolidated criteria for reporting qualitative research. Convenience and snowball sampling methods will be used to recruit firefighters to participate in the study, and interviews will be conducted using a semi-structured interview guide. The data will be analyzed in four stages using the qualitative analysis method of Krippendorff. DISCUSSION: In this study, we examine the state of health issues and healthcare usage among Korean firefighters and investigate their perceptions of and needs for integrative medicine. In this way, we aim to explore how integrative medicine and Korean medicine approaches could improve and assist healthcare services for firefighters. Furthermore, our findings will provide policymakers and healthcare providers with the necessary basic information to develop integrative medicine systems suited to firefighters.
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Bomberos , Incendios , Traumatismos Ocupacionales , Humanos , Investigación Cualitativa , República de CoreaRESUMEN
Recent tragedies around the world have shown how accidents in the cable-stayed bridges can wreak havoc on the society. To ensure the safety of the cable-stayed bridges, several studies have estimated the cable tension force using the vibration of cables. Most of these methods for estimating the tension of a cable start with measuring the displacement of the cable. Recent development of commercial cameras provide opportunity for more convenient and efficient method for measuring the displacement of cable. However, traditional vision-based displacement measurement methods require the assumption that the movement of the cable should be measured in parallel to the camera plane. This assumption limits the installation location of the camera when measuring the displacement of a cable. Therefore, this study introduces a new vision-based cable displacement measurement system that can measure the displacement of a cable in various locations even when the camera is installed in the side of the cable. The proposed method consists of three phases: (1) camera projection matrix estimation, (2) cable tracking in the image coordinate, and (3) cable displacement estimation in the world coordinate. To validate the performance of the proposed method, a simulation-based validation test, a lab-scale validation test, and an on-site validation test were conducted. The simulation-based validation test verified the performance of the proposed method in an ideal condition, and the lab-scale validation test showed the performance of the method in physical environment. Finally, the on-site validation test showed that the proposed method can measure the cable displacement with a side view camera.
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Movimiento , Vibración , Simulación por ComputadorRESUMEN
The essential actin-binding factor profilin-1 (Pfn1) is a non-classical tumor suppressor with the abilities toboth inhibit cellular proliferation and augment chemotherapy-induced apoptosis. Besides actin, Pfn1 interacts with proteins harboring the poly-L-proline (PLP) motifs. Our recent work demonstrated that both nuclear localization and PLP-binding are required for tumor growth inhibition by Pfn1, and this is at least partially due to Pfn1 association with the PLP-containing ENL protein in the Super Elongation Complex (SEC) and the transcriptional inhibition of pro-cancer genes. In this paper, by identifying a phosphorylation event of Pfn1 at Ser71 capable of inhibiting its actin-binding and nuclear export, we provide in vitro and in vivo evidence that chemotherapy-induced apoptotic sensitization by Pfn1 requires its cytoplasmic localization and actin-binding. With regard to tumor growth inhibition byPfn1, our data indicate a requirement for dynamic actin association and dissociation rendered by reversible Ser71phosphorylation and dephosphorylation. Furthermore, genetic and pharmacological experiments showed that Ser71 of Pfn1 can be phosphorylated by protein kinase A (PKA). Taken together, our data provide novel mechanistic insights into the multifaceted anticancer activities of Pfn1 and how they are spatially-defined in the cell and differentially regulated by ligand-binding.
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Aberrant expression of nuclear transporters and deregulated subcellular localization of their cargo proteins are emerging as drivers and therapeutic targets of cancer. Here, we present evidence that the nuclear exporter exportin-6 and its cargo profilin-1 constitute a functionally important and frequently deregulated axis in cancer. Exportin-6 upregulation occurs in numerous cancer types and is associated with poor patient survival. Reducing exportin-6 level in breast cancer cells triggers antitumor effects by accumulating nuclear profilin-1. Mechanistically, nuclear profilin-1 interacts with eleven-nineteen-leukemia protein (ENL) within the super elongation complex (SEC) and inhibits the ability of the SEC to drive transcription of numerous pro-cancer genes including MYC. XPO6 and MYC are positively correlated across diverse cancer types including breast cancer. Therapeutically, exportin-6 loss sensitizes breast cancer cells to the bromodomain and extra-terminal (BET) inhibitor JQ1. Thus, exportin-6 upregulation is a previously unrecognized cancer driver event by spatially inhibiting nuclear profilin-1 as a tumor suppressor.
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Carioferinas/metabolismo , Neoplasias/metabolismo , Profilinas/antagonistas & inhibidores , Profilinas/metabolismo , Animales , Línea Celular Tumoral , Femenino , Xenoinjertos , Humanos , Carioferinas/genética , Células MCF-7 , Ratones , Ratones Desnudos , Neoplasias/genética , Profilinas/genética , Análisis de Supervivencia , Regulación hacia ArribaRESUMEN
UBE2O is localized in the 17q25 locus, which is known to be amplified in human cancers, but its role in tumorigenesis remains undefined. Here we show that Ube2o deletion in MMTV-PyVT or TRAMP mice profoundly impairs tumor initiation, growth, and metastasis, while switching off the metabolic reprogramming of tumor cells. Mechanistically, UBE2O specifically targets AMPKα2 for ubiquitination and degradation, and thereby promotes activation of the mTOR-HIF1α pathway. Notably, inactivation of AMPKα2, but not AMPKα1, abrogates the tumor attenuation caused by UBE2O loss, while treatment with rapamycin or inhibition of HIF1α ablates UBE2O-dependent tumor biology. Finally, pharmacological blockade of UBE2O inhibits tumorigenesis through the restoration of AMPKα2, suggesting the UBE2O-AMPKα2 axis as a potential cancer therapeutic target.
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Proteínas Quinasas Activadas por AMP/fisiología , Neoplasias/etiología , Enzimas Ubiquitina-Conjugadoras/fisiología , Proteínas Quinasas Activadas por AMP/antagonistas & inhibidores , Animales , Antígenos de Neoplasias/metabolismo , Progresión de la Enfermedad , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/fisiología , Ratones , Ratones Endogámicos C57BL , Proteínas de Neoplasias/metabolismo , Neoplasias/tratamiento farmacológico , Serina-Treonina Quinasas TOR/fisiología , Enzimas Ubiquitina-Conjugadoras/antagonistas & inhibidores , UbiquitinaciónRESUMEN
Shikonin, which derives from Lithospermum erythrorhizon, has been traditionally used against a variety of diseases, including cancer, in Eastern Asia. Here we determined that shikonin inhibits proliferation of gastric cancer cells by inducing apoptosis. Shikonin's biological activity was validated by observing cell viability, caspase 3 activity, reactive oxygen species (ROS) generation, and apoptotic marker expressions in AGS stomach cancer cells. The concentration range of shikonin was 35-250 nM with the incubation time of 6 h. Protein levels of Nrf2 and p53 were evaluated by western blotting and confirmed by real-time PCR. Our results revealed that shikonin induced the generation of ROS as well as caspase 3-dependent apoptosis. c-Jun-N-terminal kinases (JNK) activity was significantly elevated in shikonin-treated cells, thereby linking JNK to apoptosis. Furthermore, our results revealed that shikonin induced p53 expression but repressed Nrf2 expression. Moreover, our results suggested that there may be a co-regulation between p53 and Nrf2, in which transfection with siNrf2 induced the p53 expression. We demonstrated for the first time that shikonin activated cell apoptosis in AGS cells via caspase 3- and JNK-dependent pathways, as well as through the p53-Nrf2 mediated signal pathway. Our study validates in partly the contribution of shikonin as a new therapeutic approaches/ agent for cancer chemotherapy.
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The role of AMP-activated protein kinase (AMPK) in promoting fatty acid (FA) oxidation in various tissues, such as liver and muscle, has been well understood. However, the role of AMPK in lipolysis and FA metabolism in adipose tissue has been controversial. To investigate the role of AMPK in the regulation of adipose lipolysis in vivo, we generated mice with adipose-tissue-specific knockout of both the α1 and α2 catalytic subunits of AMPK (AMPK-ASKO mice) by using aP2-Cre and adiponectin-Cre. Both models of AMPK-ASKO ablation show no changes in desnutrin/ATGL levels but have defective phosphorylation of desnutrin/ATGL at S406 to decrease its triacylglycerol (TAG) hydrolase activity, lowering basal lipolysis in adipose tissue. These mice also show defective phosphorylation of hormone-sensitive lipase (HSL) at S565, with higher phosphorylation at protein kinase A sites S563 and S660, increasing its hydrolase activity and isoproterenol-stimulated lipolysis. With higher overall adipose lipolysis, both models of AMPK-ASKO mice are lean, having smaller adipocytes with lower TAG and higher intracellular free-FA levels. Moreover, FAs from higher lipolysis activate peroxisome proliferator-activated receptor delta to induce FA oxidative genes and increase FA oxidation and energy expenditure. Overall, for the first time, we provide in vivo evidence of the role of AMPK in the phosphorylation and regulation of desnutrin/ATGL and HSL and thus adipose lipolysis.
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Proteínas Quinasas Activadas por AMP/genética , Ácidos Grasos/metabolismo , Lipasa/metabolismo , Esterol Esterasa/metabolismo , Proteínas Quinasas Activadas por AMP/metabolismo , Tejido Adiposo/metabolismo , Animales , Técnicas de Inactivación de Genes , Metabolismo de los Lípidos , Lipólisis , Ratones , Oxidación-Reducción , FosforilaciónRESUMEN
Roles of dietary phytochemicals in cancer chemoprevention via induction of nuclear factor-erythroid-2-related factor 2 (Nrf2)-mediated antioxidant enzymes have been well established in a number of studies. In this study, FACS analysis was used to reveal that the intracellular reactive oxygen species level decreased at 0-25 µM of genipin treatment. Furthermore, immunofluorescence analysis and Western blotting were used to demonstrate that genipin treatment resulted in the upregulation and nuclear translocation of Nrf2, as well as upregulation of gastrointestinal glutathione peroxidase. Finally, we found that C-Jun-NH2-kinase (JNK) was also dose-dependently activated, where depleting JNK by using a biochemical inhibitor indicated that JNK was upstream of Nrf2. Interestingly, the antioxidant effects were limited to the treatment in the lower dosage of genipin, where higher dosage of genipin treatment resulted in the increased reactive oxygen species level and cytotoxicity. Thus, this study demonstrates for the first time that lower dosage of genipin results in the induction of JNK/Nrf2/ARE signaling pathway and protection from cell death.
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Adenocarcinoma/metabolismo , Anticarcinógenos/farmacología , Iridoides/farmacología , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Factor 2 Relacionado con NF-E2/metabolismo , Neoplasias Gástricas/metabolismo , Adenocarcinoma/enzimología , Línea Celular Tumoral , Humanos , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Elementos de Respuesta , Neoplasias Gástricas/enzimologíaRESUMEN
Fatty acid and fat synthesis in the liver is a highly regulated metabolic pathway that is important for very low-density lipoprotein (VLDL) production and thus energy distribution to other tissues. Having common features at their promoter regions, lipogenic genes are coordinately regulated at the transcriptional level. Transcription factors, such as upstream stimulatory factors (USFs), sterol regulatory element-binding protein 1C (SREBP1C), liver X receptors (LXRs) and carbohydrate-responsive element-binding protein (ChREBP) have crucial roles in this process. Recently, insights have been gained into the signalling pathways that regulate these transcription factors. After feeding, high blood glucose and insulin levels activate lipogenic genes through several pathways, including the DNA-dependent protein kinase (DNA-PK), atypical protein kinase C (aPKC) and AKT-mTOR pathways. These pathways control the post-translational modifications of transcription factors and co-regulators, such as phosphorylation, acetylation or ubiquitylation, that affect their function, stability and/or localization. Dysregulation of lipogenesis can contribute to hepatosteatosis, which is associated with obesity and insulin resistance.
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Ácidos Grasos/biosíntesis , Lipogénesis/genética , Lipoproteínas VLDL/biosíntesis , Hígado/metabolismo , Transcripción Genética/genética , Animales , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice , Proteína Quinasa Activada por ADN/metabolismo , Regulación de la Expresión Génica , Lipogénesis/fisiología , Receptores X del Hígado , Ratones , Proteínas Nucleares/metabolismo , Receptores Nucleares Huérfanos/metabolismo , Proteína Quinasa C/metabolismo , Procesamiento Proteico-Postraduccional/fisiología , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Proteína 1 de Unión a los Elementos Reguladores de Esteroles/metabolismo , Serina-Treonina Quinasas TOR/metabolismo , Factores de Transcripción/metabolismo , Factores Estimuladores hacia 5'/metabolismoRESUMEN
Natural compounds are becoming important candidates in cancer therapy due to their cytotoxic effects on cancer cells by inducing various types of programmed cell deaths. In this study, we investigated whether genipin induces programmed cell deaths and mediates in Egr1/p21 signaling pathways in gastric cancer cells. Effects of genipin in AGS cancer cell lines were observed via evaluation of cell viability, ROS generation, cell cycle arrest, and protein and RNA levels of p21, Egr1, as well as apoptotic marker genes. The cell viability of AGS cells reduced by genipin treatment via induction of the caspase 3-dependent apoptosis. Cell cycle arrest was observed at the G2/M phase along with induction of p21 and p21-dependent cyclins. As an upstream mediator of p21, the transcription factor early growth response-1 (Egr1) upregulated p21 through nuclear translocation and binding to the p21 promoter site. Silencing Egr1 expression inhibited the expression of p21 and downstream molecules involved in apoptosis. We demonstrated that genipin treatment in AGS human gastric cancer cell line induces apoptosis via p53-independent Egr1/p21 signaling pathway in a dose-dependent manner.
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Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Iridoides/farmacología , Proteínas Represoras/metabolismo , Transducción de Señal/efectos de los fármacos , Neoplasias Gástricas/tratamiento farmacológico , Antineoplásicos/química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Iridoides/química , Estructura Molecular , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Relación Estructura-ActividadRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Lithospermum erythrorhizon, a naphthoquinone compound derived from a shikonin, has long been used as traditional Chinese medicine for treatment of various diseases, including cancer. To evaluate the cytotoxic effects of shikonin on AGS gastric cancer cells via induction of cell cycle arrest. MATERIALS AND METHODS: We observed the effects of 12.5-100 ng/mL dosage of shikonin treatment on AGS cancer cell line with the incubation time of 6h. Cytotoxic effects were assessed by measuring the changes in the intracellular ROS, appearance of senescence phenotype, cell cycle progression, CDK and cyclins expression levels upon shikonin treatment. We also examined upon the activation of Egr1-mediated p21 expression, by siRNA transfection, Luciferase assay, and ChIP assay. RESULTS: In this study, we found that shikonin inhibits cell proliferation by arresting cell cycle progression at the G2/M phase via modulation of p21 in AGS cells. Also, our results revealed that the p21 gene was transactivated by early growth response1 (Egr1) in response to the shikonin treatment. Transient Egr1 expression enhanced shikonin-induced p21 promoter activity, whereas the suppression of Egr1 expression by small interfering RNA attenuated the ability of shikonin to induce p21 promoter activity. CONCLUSION: Our results suggested that the anti-proliferative activity of shikonin was due to its ability to induce cell cycle arrest via Egr1-p21 signaling pathway. Thus, the work stated here validates the traditional use of shikonin in the treatment of cancer.
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Antineoplásicos/farmacología , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/genética , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Naftoquinonas/farmacología , Neoplasias Gástricas/genética , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Humanos , ARN Interferente Pequeño/genéticaRESUMEN
The bacteriophage ES2 is a virus for bacterial host cells. Unlike other phages that are known for their therapeutic effects, the ES2 phage has never been clearly examined as a therapeutic agent. To systematically and conclusively evaluate its therapeutic efficacy, the expression of the surface markers CD86, CD40, and MHCII, the production of the proinflammatory cytokines IL-6, IL-1α, IL-1ß, and TNF-α, and the underlying NF-κB signaling pathway in murine bone marrow-derived dendritic cells (BM-DCs) in response to ES2 phage infection were examined. The bacteriophage ES2, which was isolated from swine fecal samples an antigen, affected the expression of the cell surface molecules and proinflammatory cytokines that are associated with the DC maturation processes. Treatment with ES2 phage also led to NF-κBp65 activation and translocation to the nucleus, which indicates the activation of NF-κB signaling. Furthermore, the ES2 phage induced the promoter activity of IL-12p40. Our chromatin immunoprecipitation assay revealed that p65 was enriched at the IL12-p40 promoter as a direct target of chromatin. The present study demonstrates that the ES2 phage potently induces DC maturation via immune-enhancement processes.
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Bacteriófagos/inmunología , Cronobacter sakazakii/virología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Subunidad p40 de la Interleucina-12/metabolismo , FN-kappa B/metabolismo , Animales , Bacteriófagos/aislamiento & purificación , Diferenciación Celular/inmunología , Núcleo Celular/metabolismo , Citocinas/biosíntesis , Citocinas/genética , Células Dendríticas/citología , Femenino , Expresión Génica , Inmunofenotipificación , Mediadores de Inflamación/metabolismo , Subunidad p40 de la Interleucina-12/genética , Ratones , FN-kappa B/genética , Fenotipo , Transporte de Proteínas , Transducción de SeñalRESUMEN
To investigate the underlying mechanism of targets of cyanidin, a flavonoid, which exhibits potent anti-atherogenic activities in vitro and in vivo, a natural chemical library that identified potent agonistic activity between cyanidin and peroxisome proliferator-activated receptors (PPAR) was performed. Cyanidin induced transactivation activity in all three PPAR subtypes in a reporter gene assay and time-resolved fluorescence energy transfer analyses. Cyanidin also bound directly to all three subtypes, as assessed by surface plasmon resonance experiments, and showed the greatest affinity to PPARα. These effects were confirmed by measuring the expression of unique genes of each PPAR subtype. Cyanidin significantly reduced cellular lipid concentrations in lipid-loaded steatotic hepatocytes. In addition, transcriptome profiling in lipid-loaded primary hepatocytes revealed that the net effects of stimulation with cyanidin on lipid metabolic pathways were similar to those elicited by hypolipidemic drugs. Cyanidin likely acts as a physiological PPARα agonist and potentially for PPARß/δ and γ, and reduces hepatic lipid concentrations by rewiring the expression of genes involved in lipid metabolic pathways.
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Antocianinas/farmacología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , PPAR alfa/agonistas , Animales , Células CHO , Células Cultivadas , Cricetinae , Células Hep G2 , Humanos , Metabolismo de los Lípidos/efectos de los fármacos , PPAR gamma/agonistas , PPAR-beta/agonistasRESUMEN
SCOPE: A natural carotenoid abundant in seafood, astaxanthin (AX), has hypolipidemic activity, but its underlying mechanisms of action and protein targets are unknown. We investigated the molecular mechanism of action of AX in hepatic hyperlipidemia by measuring peroxisome proliferator-activated receptors (PPAR) activity. METHODS AND RESULTS: We examined the binding of AX to PPAR subtypes and its effects on hepatic lipid metabolism. AX binding activated PPAR-α, but inhibited PPAR-γ transactivation activity in reporter gene assay and time-resolved fluorescence energy transfer analyses. AX had no effect on PPARδ/ß transactivation. AX bound directly to PPAR-α and PPAR-γ with moderate affinity, as assessed by surface plasmon resonance experiments. The differential effects of AX on PPARs were confirmed by measuring the expression of unique responsive genes for each PPAR subtype. AX significantly reduced cellular lipid accumulation in lipid-loaded hepatocytes. Transcriptome analysis revealed that the net effects of stimulation with AX (100 µM) on lipid metabolic pathways were similar to those elicited by fenofibrate and lovastatin (10 µM each), with AX rewiring the expression of genes involved in lipid metabolic pathways. CONCLUSION: AX is a PPAR-α agonist and PPAR-γ antagonist, reduces hepatic lipid accumulation by rewiring the transcriptome in lipid-loaded hepatocytes.
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Hepatocitos/efectos de los fármacos , Metabolismo de los Lípidos/efectos de los fármacos , Lipotrópicos/farmacología , PPAR alfa/agonistas , PPAR gamma/antagonistas & inhibidores , Activación Transcripcional/efectos de los fármacos , Animales , Antioxidantes/metabolismo , Antioxidantes/farmacología , Células CHO , Células Cultivadas , Cricetinae , Cricetulus , Transferencia Resonante de Energía de Fluorescencia , Perfilación de la Expresión Génica , Genes Reporteros/efectos de los fármacos , Células Hep G2 , Hepatocitos/metabolismo , Humanos , Lipotrópicos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , PPAR alfa/genética , PPAR alfa/metabolismo , PPAR gamma/genética , PPAR gamma/metabolismo , Resonancia por Plasmón de Superficie , Xantófilas/metabolismo , Xantófilas/farmacologíaRESUMEN
Anthocyanins were extracted from the fruits of Rubus coreanus. Whether their antioxidant properties and antiulcer activity in gastric ulceration have been accompanied by the activation of matrix metalloproteainse-2 (MMP-2) was investigated. To assess the effect of anthocyanins on gastric ulcer, the rats were administered with anthocyanins (20, 50, and 80 mg/kg of body weight) before treatment with naproxen (80 mg/kg of body weight) to induce gastric ulceration. Lipid peroxidation and the activities of radical scavenging enzymes such as catalase, superoxide dismutase, and glutathione peroxidase were determined. The MMP-2 level was tested by zymography and Western blot. Anthocyanins of R. coreanus exhibit possible antiulcer activity in acute ulcer in a rat model by preventing lipid peroxidation and a significant increase in the activities of antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase. Also, anthocyanins induce activation of MMP-2 and attenuate the activity of the proinflammatory molecules, such as tumor necrosis factor-α and interleukin-1ß.
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Antocianinas/administración & dosificación , Metaloproteinasa 2 de la Matriz/metabolismo , Extractos Vegetales/administración & dosificación , Rosaceae/química , Úlcera Gástrica/tratamiento farmacológico , Úlcera Gástrica/enzimología , Animales , Antioxidantes/administración & dosificación , Línea Celular Tumoral , Modelos Animales de Enfermedad , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Humanos , Peroxidación de Lípido/efectos de los fármacos , Masculino , Metaloproteinasa 2 de la Matriz/genética , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/metabolismoRESUMEN
Non-steroidal anti-inflammatory drugs cause gastric ulceration through a number of mechanisms including inhibition of PG synthesis, generation of reactive oxygen species (ROS) and induction of apoptosis. Recently, matrix metalloproteinases (MMP) have been suggested to play a crucial role in these mechanisms. The present study investigated the protective effect of anthocyanins isolated from black rice bran (Heugjinjubyeo) against naproxen-induced gastric mucosal injury in rats. The oral administration of anthocyanins (5, 25 or 50 mg/kg body weight) showed significant protection against naproxen (80 mg/kg body weight)-induced gastric ulcer and inhibited lipid peroxidation in the gastric mucosa. In addition, pretreatment with anthocyanins resulted in a significant increase in the activities of radical-scavenging enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Also biochemical and zymographic analyses suggested that the administration of anthocyanins gives a significant protection against naproxen-induced gastric antral ulcer through scavenging ROS and regulation of matrix metalloproteinase-2 (MMP-2) activity. The results of intracellular radical activation show that anthocyanins suppress the generation of intracellular ROS and attenuate the suppression of MMP-2 activity by naproxen. These results suggest that anthocyanins extracted from black rice may offer potential remedy of gastric antral ulceration.
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Antocianinas/farmacología , Metaloproteinasa 2 de la Matriz/metabolismo , Úlcera Gástrica/metabolismo , Úlcera Gástrica/prevención & control , Animales , Antocianinas/aislamiento & purificación , Antiinflamatorios no Esteroideos/toxicidad , Antiulcerosos/aislamiento & purificación , Antiulcerosos/farmacología , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Femenino , Mucosa Gástrica/efectos de los fármacos , Mucosa Gástrica/metabolismo , Expresión Génica/efectos de los fármacos , Interleucina-1beta/genética , Peroxidación de Lípido/efectos de los fármacos , Metaloproteinasa 9 de la Matriz/metabolismo , Naproxeno/toxicidad , Oryza/química , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Úlcera Gástrica/inducido químicamente , Úlcera Gástrica/genética , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
This study describes a method using a DNA microarray chip to rapidly and simultaneously detect Alicyclobacillus species in orange juice based on the hybridization of genomic DNA with random probes. Three food spoilage bacteria were used in this study: Alicyclobacillus acidocaldarius, Alicyclobacillus acidoterrestris, and Alicyclobacillus cycloheptanicus. The three Alicyclobacillus species were adjusted to 2 × 10(3) CFU/ml and inoculated into pasteurized 100% pure orange juice. Cy5-dCTP labeling was used for reference signals, and Cy3-dCTP was labeled for target genomic DNA. The molar ratio of 1:1 of Cy3-dCTP and Cy5-dCTP was used. DNA microarray chips were fabricated using randomly fragmented DNA of Alicyclobacillus spp. and were hybridized with genomic DNA extracted from Bacillus spp. Genomic DNA extracted from Alicyclobacillus spp. showed a significantly higher hybridization rate compared with DNA of Bacillus spp., thereby distinguishing Alicyclobacillus spp. from Bacillus spp. The results showed that the microarray DNA chip containing randomly fragmented genomic DNA was specific and clearly identified specific food spoilage bacteria. This microarray system is a good tool for rapid and specific detection of thermophilic spoilage bacteria, mainly Alicyclobacillus spp., and is useful and applicable to the fruit juice industry.
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Alicyclobacillus/aislamiento & purificación , Bebidas/microbiología , Seguridad de Productos para el Consumidor , ADN Bacteriano/análisis , Contaminación de Alimentos/análisis , Carbocianinas , Citrus sinensis , Nucleótidos de Desoxicitosina , Microbiología de Alimentos , Humanos , Análisis por Micromatrices , Especificidad de la EspecieRESUMEN
We conducted a molecular epizootiological study of infectious bursal disease (IBD) in Korea by analyzing 85 IBD viruses (IBDVs) obtained from vaccinated or unvaccinated flocks between 1980 and 2007. Phylogenetic analysis of the partial nucleotide sequence of the hypervariable region of the VP2 gene (nucleotides 661-1020) and pathogenicity tests revealed more genetic and phenotypic diversity of IBDV in Korea than has been reported previously. We showed that very virulent IBDVs (vvIBDVs) were already present in Korea in 1986. Moreover, vvIBDVs were repeatedly detected in Korean poultry that had been vaccinated, which casts doubt on the IBD vaccine programs. We also identified novel putative antigenic variant (AV)-like IBDV isolates on the basis of their antigenic indices and the presence of amino acid changes (P222S or P222T-A321D) that are known to affect the antigenicity of VP2. These observations suggest that future studies examining the efficacy of conventional vaccines against atrophy of the bursa of Fabricius and vvIBDV shedding may be useful. Moreover, it will be of interest to determine the prevalence of putative Korean antigenic variants and whether these strains exert immunosuppressive effects in vaccinated birds.
Asunto(s)
Infecciones por Birnaviridae/veterinaria , Variación Genética , Virus de la Enfermedad Infecciosa de la Bolsa/aislamiento & purificación , Enfermedades de las Aves de Corral/epidemiología , Secuencia de Aminoácidos , Sustitución de Aminoácidos/genética , Animales , Antígenos Virales/genética , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/virología , Pollos , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Corea (Geográfico)/epidemiología , Epidemiología Molecular , Datos de Secuencia Molecular , Mutación Missense , Enfermedades de las Aves de Corral/virología , ARN Viral/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Proteínas Estructurales Virales/genéticaRESUMEN
A recombinant La Sota strain (KBNP-C4152R2L) in which fusion (F) and hemagglutinin-neuraminidase (HN) genes were replaced with those of a contemporary genotype VIId virus, KBNP-4152, has been developed. To attenuate the virulence of the recombinant strain, the F cleavage motif was mutated from (112)RRQKR(116) to (112)GRQAR(116), and to reduce pathogenic instability, a codon which does not allow changes to basic amino acids by single point mutation was inserted at codon 115. In addition a six-nucleotide sequence was inserted into the intergenic region between matrix protein and F genes for attenuation without breaking the "rule-of-six." The HN protein length was increased from 571 to 577 as a marker. Serological tests revealed that the antigenicity of KBNP-C4152R2L was similar to that of KBNP-4152 but distinct from that of the La Sota strain. KBNP-C4152R2L was avirulent (intracerebral pathogenicity index, 0.0; mean death time, >168 h) and stable in pathogenicity through in vivo passages. The killed oil emulsion of and live KBNP-C4152R2L were completely protective against mortality and egg drop caused by virulent strains, and KBNP-C4152R2L was applicable to in ovo vaccination. Therefore, KBNP-C4152R2L is a promising vaccine strain and viral vector in terms of antigenicity, productivity, safety, and pathogenic stability.
Asunto(s)
Virus de la Enfermedad de Newcastle/genética , Virus de la Enfermedad de Newcastle/inmunología , Vacunas Virales/genética , Vacunas Virales/inmunología , Sustitución de Aminoácidos/genética , Animales , Pollos , Proteína HN/genética , Datos de Secuencia Molecular , Mutagénesis Insercional , Enfermedad de Newcastle/prevención & control , ARN Viral/genética , Análisis de Secuencia de ADN , Serotipificación , Vacunas Atenuadas/genética , Vacunas Atenuadas/inmunología , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunología , Proteínas Virales de Fusión/genética , VirulenciaRESUMEN
PhiSG-JL2 is a newly discovered lytic bacteriophage infecting Salmonella enterica serovar Gallinarum biovar Gallinarum but is nonlytic to a rough vaccine strain of serovar Gallinarum biovar Gallinarum (SG-9R), S. enterica serovar Enteritidis, S. enterica serovar Typhimurium, and S. enterica serovar Gallinarum biovar Pullorum. The phiSG-JL2 genome is 38,815 bp in length (GC content, 50.9%; 230-bp-long direct terminal repeats), and 55 putative genes may be transcribed from the same strand. Functions were assigned to 30 genes based on high amino acid similarity to known proteins. Most of the expected proteins except tail fiber (31.9%) and the overall organization of the genomes were similar to those of yersiniophage phiYeO3-12. phiSG-JL2 could be classified as a new T7-like virus and represents the first serovar Gallinarum biovar Gallinarum phage genome to be sequenced. On the basis of intraspecific ratios of nonsynonymous to synonymous nucleotide changes (Pi[a]/Pi[s]), gene 2 encoding the host RNA polymerase inhibitor displayed Darwinian positive selection. Pretreatment of chickens with phiSG-JL2 before intratracheal challenge with wild-type serovar Gallinarum biovar Gallinarum protected most birds from fowl typhoid. Therefore, phiSG-JL2 may be useful for the differentiation of serovar Gallinarum biovar Gallinarum from other Salmonella serotypes, prophylactic application in fowl typhoid control, and understanding of the vertical evolution of T7-like viruses.