RESUMEN
A simultaneous analytical method was developed for the determination of alkyl furans (Furan, 2-methylfuran, 3-methylfuran and 2,5-dimethylfuran) in processed foods by headspace-GC-MS. Single-laboratory validation data of furan, 2-methylfuran, 3-methylfuran and 2,5-dimethylfuran showed good precision and accuracy. The mean recoveries ranged from 92 to 116%, the intermediate precision (RSDi) ranged from 0.9 to 12.9%. The level of LOQ ranged from 0.5 to 1.2 µg/kg (coffee), from 3.5 to 4.1 µg/kg (soy sauce), from 0.4 to 1.3 µg/kg (other foods: clear apple juice, infant formula and baby food), respectively. This method has the sensitivity to detect low levels of furan and alkyl furans contaminated in various foods and is thus applicable to surveillance for risk management in food safety.
Asunto(s)
Alimentos Procesados , Furanos , Lactante , Humanos , Café , Inocuidad de los AlimentosRESUMEN
Single-walled carbon nanotubes (SWCNTs) have the potential to revolutionize nanoscale electronics and power sources; however, their low purity and high separation cost limit their use in practical applications. Here we present a supramolecular chemistry-based one-pot, less expensive, scalable, and highly efficient separation of a solubilizer/adsorbent-free pure semiconducting SWCNT (sc-SWCNT) using flavin/isoalloxazine analogues with different substituents. On the basis of both experimental and computational simulations (DFT study), we have revealed the molecular requirements of the solubilizers as well as provided a possible mechanism for such a highly efficient selective sc-SWCNT separation. The present sorting method is very simple (one-pot) and gives a promising sc-SWCNT separation methodology. Thus, the study provides insight for the molecular design of an sc-SWCNT solubilizer with a high (n,m)-chiral selectivity, which benefits many areas including semiconducting nanoelectronics, thermoelectric, bio and energy materials, and devices using solubilizer-free very pure sc-SWCNTs.
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Noncoding (nc) RNA called satellite I is transcribed from the human centromere region. Depletion of this ncRNA results in abnormal nuclear morphology because of defects in chromosome segregation. Some protein factors interact with this ncRNA and function as a component of a nc ribonucleoprotein (RNP) complex in mitotic regulation. Here, we found that DHX38, a pre-mRNA splicing-related DEAH box RNA helicase, interacts with satellite I ncRNA. Depletion of DHX38 resulted in defective chromosome segregation similar to knockdown of satellite I ncRNA. Interaction between DHX38 and ncRNA was interphase-specific, but DHX38 depletion affected the function of Aurora B, which associated with satellite I ncRNA at mitotic phase. Based on these findings, we suggest that DHX38 has a role in mitotic regulation as a component of the satellite I ncRNP complex at interphase.
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Centrómero/genética , Segregación Cromosómica , ARN Helicasas DEAD-box/metabolismo , ADN Satélite , Factores de Empalme de ARN/metabolismo , ARN no Traducido/genética , Línea Celular , Técnicas de Silenciamiento del Gen , HumanosRESUMEN
Human centromeres consist of repetitive sequences from which satellite I noncoding RNAs are transcribed. We found that knockdown of satellite I RNA causes abnormal chromosome segregation and generation of nuclei with a grape-shape phenotype. Co-immunoprecipitation experiments showed that satellite I RNA associates with Aurora B, a component of the chromosome passenger complex (CPC) regulating proper attachment of microtubules to kinetochores, in mitotic HeLa cells. Satellite I RNA was also shown to associate with INCENP, another component of the CPC. In addition, depletion of satellite I RNA resulted in up-regulation of kinase activity of Aurora B and delocalization of the CPC from the centromere region. These results suggest that satellite I RNA is involved in chromosome segregation through controlling activity and centromeric localization of Aurora B kinase.
Asunto(s)
Segregación Cromosómica/fisiología , ARN no Traducido/metabolismo , Aurora Quinasa B/genética , Aurora Quinasa B/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , División del Núcleo Celular , Centrómero/genética , Centrómero/metabolismo , Proteínas Cromosómicas no Histona/genética , Proteínas Cromosómicas no Histona/metabolismo , Células HeLa , Humanos , InterfaseRESUMEN
OBJECTIVE: Accumulating evidences suggest that cancer-associated fibroblasts are provided from bone-marrow-derived mesenchymal stem cells (BM-MSCs); however, little is known about the mechanism(s) by which BM-MSCs accelerate cancer aggressiveness. METHODS: Gastric carcinoma (GC)-derived MKN-7 cells were cocultured with UE6E7T-12 BM-MSCs. The gene expression profile in MKN-7 cells was investigated by microarray analysis. Between two major types of GCs (intestinal- and diffuse-type), the expression of genes was detected by immunohistochemistry. RESULTS: We found that direct attachment to UE6E7T-12 induced proliferation and cluster formation of MKN-7 cells. Coculture with UE6E7T-12 increased the population of CD133+ MKN-7 cells in vitro and coimplantation of these in mice resulted in subcutaneous tumors in vivo. The wingless-type MMTV integration site (WNT) family member 5A (WNT5A) and transforming growth factor-ß (TGF-ß)-induced (TGFBI) genes were found to be upregulated in MKN-7 cells directly attached to UE6E7T-12. Recruitment of CD271+ BM-MSC was detected preferentially in the stroma of the diffuse-type GC and this type of GC cell also showed frequent expression of WNT5A, TGF-ß type I receptor and CD133. CONCLUSION: BM-MSC-mediated activations of the WNT and TGF-ß signaling pathways were thought to provide advantageous microenvironments for cancer progression by supporting the reacquisition and maintenance of cancer stem cells.
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Células Madre Mesenquimatosas/fisiología , Células Madre Neoplásicas/fisiología , Factor de Crecimiento Transformador beta/genética , Microambiente Tumoral , Proteínas Wnt/genética , Adulto , Animales , Línea Celular Tumoral , Proliferación Celular , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , Masculino , Ratones , Ratones SCID , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Regulación hacia Arriba/genética , Proteína Wnt-5aRESUMEN
Recent popularization of next-generation sequencing enables conducting easy transcriptome analysis. Nevertheless, substantial RNA isolation work prior to RNA sequencing, as well as the high cost involved, still makes the routine use of large-scale transcriptome analysis difficult. For example, conventional phenol-chloroform RNA extraction cannot be easily applied to hundreds of samples. Therefore, we developed Direct-TRI, a new cost-effective and high throughput RNA-extraction method that uses a commercial guanidine-phenol-based RNA extraction reagent and a 96-well silica column plate. We applied Direct-TRI to zebrafish whole larvae and juvenile samples and obtained comparable RNA qualities by several different homogenization methods such as vortexing, manual homogenizing, and freezing/crushing. Direct-TRI enabled the extraction of 192 RNA samples in an hour with a cost of less than a dollar per sample. Direct-TRI is useful for large-scale transcriptome studies, manipulating hundreds of zebrafish individuals, and may be used with other animal samples.
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The amphiphilic alpha-helical peptide, Td3717, is a bi-functional synthetic peptide that acts as both a polycation for DNA binding and a ligand for targeted delivery to tumor cells. Td3717 forms a stable complex with plasmid DNA, and the complex maintained high transfection efficiency after storage at 4 degrees C for six months and after four freeze/thaw cycles. During the storage and freeze/thaw cycling, the particle size of the DNA/Td3717 complex remained less than 100nm. The size of the complex is an important factor for its internalization into cells via the endocytosis pathway; therefore, the stability of the particles will strongly contribute to high transfection efficiencies after storage and repeated freezing/thawing.
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Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , ADN/química , ADN/metabolismo , Portadores de Fármacos/química , Portadores de Fármacos/metabolismo , Transfección/métodos , Secuencia de Aminoácidos , Animales , Proteínas Portadoras/síntesis química , Proteínas Portadoras/genética , Células Cultivadas , Chlorocebus aethiops , ADN/genética , Endocitosis , Congelación , Haplorrinos , Datos de Secuencia Molecular , Estructura Secundaria de Proteína , Manejo de Especímenes , Células VeroRESUMEN
Although the Apiaceae herb family has been traditionally used for the management of type 2 diabetes, its molecular mechanism has not been clarified. Coumarin derivatives, which are abundant in plants of the Apiaceae family, were evaluated for their effects on adipogenesis. We found that suksdorfin significantly promoted adipocyte differentiation and enhanced production of adiponectin, an anti-diabetic adipokine. We also demonstrated that suksdorfin activates peroxisome proliferator-activated receptor gamma (PPARγ), a master regulator of adipogenesis. Furthermore, we showed metabolic disorders in obese diabetic KK-Ay mice were attenuated by suksdorfin feeding. Suksdorfin intake induced adipocyte miniaturization and increased expression levels of PPARγ target genes related to adipocyte differentiation. These results indicated that suksdorfin induces adipogenesis in white adipose tissue (WAT) via the activation of PPARγ, leading to improvement of obesity-induced metabolic disorders. Therefore, suksdorfin-mediated amelioration of WAT dysfunctions might be responsible for the anti-diabetic effects of traditional herbal medicine therapy with Apiaceae.
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Adipocitos/efectos de los fármacos , Cumarinas/administración & dosificación , Trastornos del Metabolismo de la Glucosa/tratamiento farmacológico , PPAR gamma/metabolismo , Células 3T3-L1 , Adipocitos/citología , Adipocitos/metabolismo , Adiponectina/metabolismo , Animales , Apiaceae/química , Diferenciación Celular/efectos de los fármacos , Cumarinas/farmacología , Activación Enzimática/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Trastornos del Metabolismo de la Glucosa/enzimología , Ratones , Ratones Obesos , Transducción de Señal/efectos de los fármacosRESUMEN
Dehydroabietic acid (DAA) is a food-derived terpenoid with various bioactivities. Our previous study has revealed that DAA activates peroxisome proliferator-activated receptor-γ (PPARγ) in luciferase assay and suppresses chronic inflammation in obese adipose tissues. In this study, we examined the effects of DAA on adipocyte differentiation. DAA treatment stimulated the adipocyte differentiation of 3T3-L1 preadipocytes. The DAA treatment increased the mRNA expression levels of adipocyte differentiation marker genes such as aP2, lipoprotein lipase (LPL), and PPARγ. In particular, the expression level of adiponectin, which is an adipocytokine with stimulatory effects on insulin sensitivity, was increased at both the mRNA and protein levels by the DAA treatment. Moreover, the DAA treatment stimulated insulin-dependent glucose uptake into differentiated 3T3-L1 adipocytes. These findings indicate that DAA stimulates adipocyte differentiation and insulin sensitivity in 3T3-L1 cells, suggesting that DAA is a valuable food-derived compound for the management of metabolic syndrome.
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Abietanos/farmacología , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Células 3T3-L1 , Adipocitos/citología , Animales , Diferenciación Celular/efectos de los fármacos , Proteínas de Unión a Ácidos Grasos/genética , Lipoproteína Lipasa/genética , Ratones , PPAR gamma/genética , ARN Mensajero/genéticaRESUMEN
A cationic peptide, Td3701, which was derived from factor VIII that has affinity with phosphatidylserine (PS), showed efficient transfection ability for cells that express PS on the cell surface. PS is exposed on tumor cell surfaces therefore we have focused on PS as the target molecule for tumor specific gene delivery. In this article, to improve transfection efficiency and specificity in targeting tumor cells, some amino acid residues of Td3701 were replaced. The resulting peptide, Td3717, shows higher transfection efficiency (more than 30 times that of Td3701). The transfection efficiency was dependent on the amount of PS on the cell surface, suggesting that Td3717 bound with plasmid DNA could recognize PS on the cell surface. Td3717 is expected to be useful as an efficient gene carrier molecule specific to PS-presenting tumor cells.
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Técnicas de Transferencia de Gen , Vectores Genéticos/metabolismo , Péptidos/química , Péptidos/metabolismo , Fosfatidilserinas/metabolismo , Secuencia de Aminoácidos , Animales , Línea Celular Tumoral , Chlorocebus aethiops , Vectores Genéticos/genética , Humanos , Datos de Secuencia Molecular , Péptidos/síntesis química , Péptidos/genética , Células VeroRESUMEN
Interaction between adipocytes and macrophages contributes to the development of insulin resistance in obese adipose tissues. In this study, we examined whether luteolin, food-derived flavonoid, could suppress the production of inflammatory mediators of the interaction between adipocytes and macrophages. Experiments using a coculture system of adipocytes and macrophages showed that luteolin suppressed the production of inflammatory mediators. In addition, activated macrophages were targets for the suppressive effect of luteolin. Luteolin inhibited the phosphorylation of JNK and suppressed the production of inflammatory mediators in the activated macrophages. The findings indicate that luteolin can inhibit the interaction between adipocytes and macrophages to suppress the production of inflammatory mediators, suggesting that luteolin is a valuable food-derived compound for the treatment of metabolic syndrome.
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Adipocitos/efectos de los fármacos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , Luteolina/farmacología , Macrófagos/efectos de los fármacos , Células 3T3-L1 , Adipocitos/citología , Adipocitos/metabolismo , Animales , Western Blotting , Comunicación Celular/efectos de los fármacos , Línea Celular , Técnicas de Cocultivo , Medios de Cultivo Condicionados/farmacología , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Flavonoides/farmacología , Alimentos , Proteínas I-kappa B/metabolismo , Mediadores de Inflamación/metabolismo , Macrófagos/citología , Macrófagos/metabolismo , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
PURPOSE: The aim of this study was to measure the volume of each pulmonary segment by volumetric computed tomography (CT) data using a newly developed three-dimensional software application and to identify the differences between those with chronic obstructive pulmonary disease (COPD) and controls. MATERIALS AND METHODS: CT scans of 11 COPD patients and 16 controls were included. The volume of each pulmonary segment was measured by each of two operators to evaluate the reproducibility of the software. This measured volume was then divided by the total lung volume to revise individual variations. RESULTS: Volumes of the right (rt) S2, rt S5, left (lt) S1 + S2, lt S3, and lt S5 were significantly larger in COPD patients than in controls (P < 0.05). Regarding the ratio of the volume of each pulmonary segment per total lung volume, the areas of rt S2 and lt S1 + S2 were significantly larger in COPD patients than in controls (P < 0.05), whereas lt S10 was significantly smaller in COPD patients than in controls (P < 0.05). CONCLUSION: We measured the volume of each pulmonary segment based on volumetric CT data using this software. In addition, we demonstrated that the upper lung volume of COPD subjects was larger than that of controls, whereas the lower lung volumes were almost the same.
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Imagenología Tridimensional , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico por imagen , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Programas Informáticos , Tomografía Computarizada por Rayos X , Adulto , Anciano , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Reproducibilidad de los Resultados , Pruebas de Función Respiratoria , Estadísticas no ParamétricasRESUMEN
Since phosphatidylserine (PS) is known to translocate to the external face of the plasma membrane when the cell membrane becomes disordered, we decided to focus our attention on PS as a target molecule for gene delivery. In this paper, the novel peptide Td3701 was designed, synthesized, and characterized for its physico-chemico-biological properties. Td3701 simultaneously exhibited both characters as a DNA carrier and a sensor probe for active targeting, which seemed to be triggered by structural changes in the presence of PS. This is a very unique character among nonviral vectors, and it is believed that Td3701 could be used for selective gene delivery.