RESUMEN
In this study, we were challenging to identify characteristic compounds in breast cancer cell lines. GC analysis of extracts from the culture media of breast cancer cell lines (MCF-7, SK-BR-3, and YMB-1) using a solid-phase Porapak Q extraction revealed that two compounds of moderate volatility, 1-hexadecanol and 5-(Z)-dodecenoic acid, were detected with markedly higher amount than those in the medium of fibroblast cell line (KMST-6). Furthermore, LC-TOF/MS analysis of the extracts clarified that in addition to the above two fatty acids, the amounts of five unsaturated fatty acids [decenoic acid (C10:1), decadienoic acid (C10:2), 5-(Z)-dodecenoic acid (C12:1), 5-(Z)-tetradecenoic acid (C14:1), and tetradecadienoic acid (C14:2)] in MCF-7 medium were higher than those in medium of KMST-6. Interestingly, H2O2-oxidation of 5-(Z)-dodecenoic acid and 5-(Z)-tetradecenoic acid produced volatile aldehydes that were reported as specific volatiles in breath from various cancer patients, such as heptanal, octanal, nonanal, decanal, 2-(E)-nonenal, and 2-(E)-octenal. Thus, we concluded that these identified compounds over-produced in breast cancer cells in this study could serve as potential precursors producing reported cancer-specific volatiles.
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Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Ácidos Grasos/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Ácidos Grasos/análisis , Femenino , Cromatografía de Gases y Espectrometría de Masas , Humanos , Oxidación-Reducción , Microextracción en Fase Sólida , Células Tumorales Cultivadas , Compuestos Orgánicos Volátiles/análisisRESUMEN
The COPII-cargo adaptor complex Lst1-Sec23 selectively sorts proteins into vesicles that bud from the endoplasmic reticulum (ER) and traffic to the Golgi. Improperly folded proteins are prevented from exiting the ER and are degraded. ER-phagy is an autophagic degradation pathway that uses ER-resident receptors. Working in yeast, we found an unexpected role for Lst1-Sec23 in ER-phagy that was independent from its function in secretion. Up-regulation of the stress-inducible ER-phagy receptor Atg40 induced the association of Lst1-Sec23 with Atg40 at distinct ER domains to package ER into autophagosomes. Lst1-mediated ER-phagy played a vital role in maintaining cellular homeostasis by preventing the accumulation of an aggregation-prone protein in the ER. Lst1 function appears to be conserved because its mammalian homolog, SEC24C, was also required for ER-phagy.
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Autofagia , Vesículas Cubiertas por Proteínas de Revestimiento/metabolismo , Retículo Endoplásmico/metabolismo , Proteínas Activadoras de GTPasa/metabolismo , Proteínas de la Membrana/metabolismo , Proteolisis , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Proteínas Relacionadas con la Autofagia/metabolismo , Estrés del Retículo Endoplásmico , Agregado de Proteínas , Agregación Patológica de Proteínas/metabolismo , Receptores Citoplasmáticos y Nucleares/metabolismo , Respuesta de Proteína DesplegadaRESUMEN
Regulation of plasma membrane (PM) protein abundance by selective endocytosis is critical for cellular adaptation to stress or changing nutrient availability. One example involves rapid endocytic turnover of Mup1, a yeast methionine transporter, in response to increased methionine availability. Here, we report that methionine triggers rapid translocation of the ubiquitin ligase adaptor Art1 to the PM and dephosphorylation of Art1 at specific threonine residues. This methionine-induced dephosphorylation of Art1 is mediated by Ppz phosphatases, and analysis of phosphomimetic and phosphorylation-defective variants of Art1 indicates that these events toggle Art1 recognition of Mup1 at the PM. Importantly, we find that Ppz phosphatases are dispensable for Art1 PM translocation, but are required for Art1 interaction with Mup1. Based on our findings, we propose that methionine influx triggers Art1 translocation to the PM, followed by Ppz-mediated dephosphorylation which promotes cargo recognition at the PM.
Asunto(s)
Membrana Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Endocitosis/fisiología , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Factores de Transcripción/metabolismo , Membrana Celular/genética , Proteínas de Unión al ADN/genética , Fosforilación/fisiología , Proteínas/genética , Proteínas/metabolismo , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Factores de Transcripción/genéticaRESUMEN
Amorphous Si (a-Si) films with metal silicide are expected to enhance the absorption ability of pure a-Si films. In this present study, NiSi (20 nm)/Si (40 nm) and AlSi (20 nm)/Si (40 nm) bilayer thin films are deposited through radio frequency (RF) sputtering at room temperature. The influence of the film's composition and the annealing temperature on the film's optical absorption is investigated. The results show that all the NiSi/Si films and AlSi/Si films possess higher absorption ability compared to a pure a-Si film (60 nm). After annealing from 400 to 600 °C under vacuum for 1 h, the Si layer remains amorphous in both NiSi/Si films and AlSi/Si films, while the NiSi layer crystallizes into NiSi2 phase, whereas Al atoms diffuse through the whole film during the annealing process. Consequently, with increasing the annealing temperature, the optical absorption of NiSi/Si films increases, while that of AlSi/Si films obviously degrades.
RESUMEN
Endocytic down-regulation of cell-surface proteins is a fundamental cellular process for cell survival and adaptation to environmental stimuli. Ubiquitination of cargo proteins serves as the sorting signal for downstream trafficking and relies on the arrestin-related trafficking adaptor (ART)-Rsp5 ubiquitin ligase adaptor network in yeast. Hence proper regulation of the abundance and activity of these ligase-adaptor complexes is critical for main-tenance of optimal plasma membrane protein composition. Here we report that the stability of ARTs is regulated by the deubiquitinating enzymes (DUBs) Ubp2 and Ubp15. By counteracting the E3 ubiquitin ligase Rsp5, Ubp2 and Ubp15 prevent hyperubiquitination and proteasomal degradation of ARTs. Specifically, we show that loss of both Ubp2 and Ubp15 results in a defect in Hxt6 endocytosis associated with Art4 instability. Our results uncover a novel function for DUBs in the endocytic pathway by which Ubp2 and Ubp15 positively regulate the ART-Rsp5 network.
Asunto(s)
Enzimas Desubicuitinizantes/metabolismo , Enzimas Desubicuitinizantes/fisiología , Endopeptidasas/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Arrestinas/metabolismo , Membrana Celular/metabolismo , Endocitosis/fisiología , Complejos de Clasificación Endosomal Requeridos para el Transporte/metabolismo , Unión Proteica , Transporte de Proteínas , Saccharomyces cerevisiae/metabolismo , Ubiquitina/metabolismo , Complejos de Ubiquitina-Proteína Ligasa/metabolismo , Ubiquitina-Proteína Ligasas/metabolismo , Ubiquitinación/fisiologíaRESUMEN
In this study, we reported the synthesis of the two-dimensional (2D) nanocomposite of molybdenum disulfide and nitrogen-doped graphene oxide (MoS2/nGO) as a platinum-free counter electrode (CE) for dye-sensitized solar cells (DSSCs). X-ray photoelectron spectroscopy (XPS), high-resolution transmission electron microscopy (HRTEM), and Raman spectroscopy were used to examine the characteristics of the 2D nanocomposite of MoS2/nGO. The cyclic voltammetry (CV), electrochemical impedance spectra (EIS), and the Tafel polarization measurements were carried out to examine the electrocatalytic abilities. XPS and Raman results showed the 2D behaviors of the prepared nanomaterials. HRTEM micrographs showed the direct evidence of the 2D nanocomposite of MoS2/nGO. The results of electrocatalytic examinations indicated the MoS2/nGO owning the low charge transfer resistance, high electrocatalytic activity, and fast reaction kinetics for the reduction of triiodide to iodide on the electrolyte-electrode interface. The 2D nanocomposite of MoS2/nGO combined the advantages of the high specific surface of nGO and the plenty edge sites of MoS2 and showed the promoted properties different from those of their individual constituents to create a new outstanding property. The DSSC with MoS2/nGO nanocomposite CE showed a photovoltaic conversion efficiency (PCE) of 5.95 % under an illumination of AM 1.5 (100 mW/cm(2)), which was up to 92.2 % of the DSSC with the conventional platinum (Pt) CE (PCE = 6.43 %). These results reveal the potential of the MoS2/nGO nanocomposite in the use of low-cost, scalable, and efficient Pt-free CEs for DSSCs.
RESUMEN
Using density functional theory and the Hubbard U method, we investigated the geometric structure, electronic structure, and optical property of Al/Ga-codoped ZnO. A 3 × 3 × 3 ZnO supercell was used to construct Al- and Ga-monodoped ZnO structures and Al/Ga-codoped ZnO (AGZO) structures. All three structures showed n-type conduction, and the optical band gaps were larger than that of pure ZnO. For a given impurity concentration, Ga impurities contribute more free carriers than Al impurities in AGZO. However, the presence of Al impurities improves the transmittance. These results can theoretically explain the factors that influence the electrical and optical properties.
RESUMEN
We systematically investigated the effects of Al-impurity type on the formation energy, crystal structure, charge density, electronic structure, and optical properties of ZnO by using density functional theory and the Hubbard-U method. Al-related defects, such as those caused by the substitution of Zn and O atoms by Al atoms (Als(Zn) and Als(O), respectively) and the presence of an interstitial Al atom at the center of a tetrahedron (Ali(tet)) or an octahedron (Ali(oct)), and various Al concentrations were evaluated. The calculated formation energy follows the order Ef(Als(Zn)) < Ef(Ali(tet)) < Ef(Ali(oct)) < Ef(Als(O)). Electronic structure analysis showed that the Als(Zn), Als(O), Ali(tet), and Ali(oct) models follow n-type conduction, and the optical band gaps are higher than that of pure ZnO. The calculated carrier concentrations of the Als(O) and Ali(tet)/Ali(oct) models are higher than that of the Als(Zn) model. However, according to the curvature of the band structure, the occurrence of interstitial Al atoms or the substitution of O atoms by Al atoms results in a high effective mass, possibly reducing the carrier mobility. The average transmittance levels in the visible light and ultraviolet (UV) regions of the Als(Zn) model are higher than those of pure ZnO. However, the presence of an interstitial Al atom within the ZnO crystal reduces transmittance in the visible light region; Als(O) substantially reduces the transmittance in the visible light and UV regions. In addition, the properties of ZnO doped with various Als(Zn) concentrations were analyzed.
RESUMEN
Platinum nanocubes (PtNCs) were deposited onto a fluorine-doped tin oxide glass by electrochemical deposition (ECD) method and utilized as a counter electrode (CE) for dye-sensitized solar cells (DSSCs). In this study, we controlled the growth of the crystalline plane to synthesize the single-crystal PtNCs at room temperature. The morphologies and crystalline nanostructure of the ECD PtNCs were examined by field emission scanning electron microscopy and high-resolution transmission electron microscopy. The surface roughness of the ECD PtNCs was examined by atomic force microscopy. The electrochemical properties of the ECD PtNCs were analyzed by cyclic voltammetry, Tafel polarization, and electrochemical impedance spectra. The Pt loading was examined by inductively coupled plasma mass spectrometry. The DSSCs were assembled via an N719 dye-sensitized titanium dioxide working electrode, an iodine-based electrolyte, and a CE. The photoelectric conversion efficiency (PCE) of the DSSCs with the ECD PtNC CE was examined under the illumination of AM 1.5 (100 mWcm(-2)). The PtNCs in this study presented a single-crystal nanostructure that can raise the electron mobility to let up the charge-transfer impedance and promote the charge-transfer rate. In this work, the electrocatalytic mass activity (MA) of the Pt film and PtNCs was 1.508 and 4.088 mAmg(-1), respectively, and the MA of PtNCs was 2.71 times than that of the Pt film. The DSSCs with the pulse-ECD PtNC CE showed a PCE of 6.48 %, which is higher than the cell using the conventional Pt film CE (a PCE of 6.18 %). In contrast to the conventional Pt film CE which is fabricated by electron beam evaporation method, our pulse-ECD PtNCs maximized the Pt catalytic properties as a CE in DSSCs. The results demonstrated that the PtNCs played a good catalyst for iodide/triiodide redox couple reactions in the DSSCs and provided a potential strategy for electrochemical catalytic applications.
RESUMEN
A two-dimensional nanostructure of molybdenum disulfide (MoS2) thin film exposed layered nanosheet was prepared by a low-temperature thermally reduced (TR) method on a fluorine-doped tin oxide (FTO) glass substrate as a platinum (Pt)-free and highly electrocatalytic counter electrode (CE) for dye-sensitized solar cells (DSSCs). Thermogravimetric analysis (TGA) results show that the MoS2 sulfidization temperature was approximately 300 °C. X-ray photoelectron spectroscopy (XPS), high-resolution transmission electron microscopy (HRTEM), and X-ray diffraction (XRD) indicate that the stoichiometry and crystallization of MoS2 were more complete at higher temperatures; however, these temperatures reduce the number of edge-plane active sites in the short-range-order nanostructure. Accordingly, the DSSCs with 300 °C annealed TR-MoS2 CE exhibited an excellent photovoltaic conversion efficiency (PCE) of 6.351 %, up to 91.7 % of which is obtained using the conventional TD-Pt CE (PCE = 6.929 %). The temperature of thermal reaction and the molar ratio of reaction precursors were found to significantly influence the resulting stoichiometry and crystallization of MoS2 nanosheets, thus affecting DSSCs' performance.
RESUMEN
Proper segregation of chromosomes during meiosis requires the formation and repair of double-strand breaks (DSBs) to form crossovers. Repair is biased toward using the homolog as a substrate rather than the sister chromatid. Pch2 is a conserved member of the AAA(+)-ATPase family of proteins and is implicated in a wide range of meiosis-specific processes including the recombination checkpoint, maturation of the chromosome axis, crossover control, and synapsis. We demonstrate a role for Pch2 in promoting and regulating interhomolog bias and the meiotic recombination checkpoint in response to unprocessed DSBs through the activation of axial proteins Hop1 and Mek1 in budding yeast. We show that Pch2 physically interacts with the putative BRCT repeats in the N-terminal region of Xrs2, a member of the MRX complex that acts at sites of unprocessed DSBs. Pch2, Xrs2, and the ATM ortholog Tel1 function in the same pathway leading to the phosphorylation of Hop1, independent of Rad17 and the ATR ortholog Mec1, which respond to the presence of single-stranded DNA. An N-terminal deletion of Xrs2 recapitulates the pch2Δ phenotypes for signaling unresected breaks. We propose that interaction with Xrs2 may enable Pch2 to remodel chromosome structure adjacent to the site of a DSB and thereby promote accessibility of Hop1 to the Tel1 kinase. In addition, Xrs2, like Pch2, is required for checkpoint-mediated delay conferred by the failure to synapse chromosomes.
Asunto(s)
Cromosomas Fúngicos/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Meiosis/genética , Proteínas Nucleares/genética , Proteínas Serina-Treonina Quinasas/genética , Proteínas de Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/genética , Transportadoras de Casetes de Unión a ATP/genética , Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Proteínas de Ciclo Celular/genética , Emparejamiento Cromosómico , Segregación Cromosómica/genética , Intercambio Genético/genética , Roturas del ADN de Doble Cadena , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Puntos de Control de la Fase G2 del Ciclo Celular , MAP Quinasa Quinasa 1/genética , Fosforilación , Secuencias Repetidas TerminalesRESUMEN
BACKGROUND: Homologous recombination promotes proper segregation of chromosomes during meiosis. Programmed double-strand breaks (DSBs) initiate recombination and are repaired preferentially using the homolog rather than the sister chromatid template. In yeast, activation of Mek1 kinase upholds this bias. Mek1 is also a proposed effector kinase in the recombination checkpoint that responds to aberrant DNA and/or axis structures. Elucidating a role for Mek1 in this checkpoint has been difficult, because a mek1 null mutation causes rapid repair of DSBs using a sister chromatid, thus bypassing formation of checkpoint-activating lesions. Here we analyzed a MEK1 gain-of-function allele to test if it would enhance interhomolog bias and/or the checkpoint response. RESULTS: When Mek1 activation was artificially maintained through glutathione S-transferase-mediated dimerization, there was an enhanced skew toward interhomolog recombination and reduction of intersister events, including multichromatid joint molecules. Increased interhomolog events were specifically repaired as noncrossovers rather than as crossovers. Ectopic Mek1 dimerization was also sufficient to impose interhomolog bias in the absence of recombination checkpoint functions, thereby uncoupling these two processes. Finally, the stringency of the checkpoint response was enhanced in mutants with weak recombination defects by blocking prophase exit in a subset of cells in which arrest is not absolute. CONCLUSIONS: We propose that Mek1 plays dual roles during meiotic prophase I by phosphorylating targets directly involved in the recombination checkpoint, as well as targets involved in sister chromatid recombination. We discuss how regulation of pachytene exit by Mek1 or similar kinases could influence checkpoint stringency, which may differ among species and between sexes.
Asunto(s)
Ciclo Celular , MAP Quinasa Quinasa 1/metabolismo , Meiosis/genética , Recombinación Genética , Alelos , Roturas del ADN de Doble Cadena , Reparación del ADN , Genes cdc , MAP Quinasa Quinasa 1/química , MAP Quinasa Quinasa 1/genética , Profase Meiótica I/fisiología , Multimerización de Proteína , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Saccharomyces cerevisiae/citología , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMEN
Death-associated protein kinase (DAPK) was identified as a mediator of interferon (IFN)-induced cell death. How IFN controls DAPK activation remains largely unknown. Here, we identify the BTB-Kelch protein KLHL20 as a negative regulator of DAPK. KLHL20 binds DAPK and Cullin 3 (Cul3) via its Kelch-repeat domain and BTB domain, respectively. The KLHL20-Cul3-ROC1 E3 ligase complex promotes DAPK polyubiquitination, thereby inducing the proteasomal degradation of DAPK. Accordingly, depletion of KLHL20 diminishes DAPK ubiquitination and degradation. The KLHL20-mediated DAPK ubiquitination is suppressed in cells receiving IFN-alpha or IFN-gamma, which induces an enrichment/sequestration of KLHL20 in the PML nuclear bodies, thereby separating KLHL20 from DAPK. Consequently, IFN triggers the stabilization of DAPK. This mechanism of DAPK stabilization is crucial for determining IFN responsiveness of tumor cells and contributes to IFN-induced autophagy. This study identifies KLHL20-Cul3-ROC1 as an E3 ligase for DAPK ubiquitination and reveals a regulatory mechanism of DAPK, through blocking its accessibility to this E3 ligase, in IFN-induced apoptotic and autophagic death. Our findings may be relevant to the problem of IFN resistance in cancer therapy.
Asunto(s)
Proteínas Portadoras/química , Proteínas Cullin/química , Regulación de la Expresión Génica , Interferones/química , Ubiquitina/química , Proteínas Adaptadoras Transductoras de Señales , Animales , Proteínas Reguladoras de la Apoptosis/metabolismo , Autofagia , Proteínas Quinasas Dependientes de Calcio-Calmodulina/metabolismo , Proteínas Quinasas Asociadas a Muerte Celular , Células HeLa , Humanos , Interferones/metabolismo , Ratones , Modelos Biológicos , Células 3T3 NIH , Neoplasias/terapia , Fenotipo , Ubiquitina-Proteína Ligasas/metabolismoRESUMEN
This case report describes hospice care provided to a patient suffering from terminal stage liver cancer. Care was analyzed based on the Neuman System Model. In the nursing process, the author first established a good relationship with the patient and then collected information related to patient physical, mental and spiritual needs through discussions, observations and anamnesis. Results revealed that sources of pressure affecting patient functional system harmony came from two intrinsic factors; namely, intrapersonal and interpersonal stressors. Intrapersonal stressors included changes in nutritional status, aches, and struggles with fear of death. Interpersonal stressors addressed the guilt of being unable to meet the expectations of family members. Based on the Neuman System Model, patient pressures were analyzed at three levels of prevention in order to strengthen their ability to cope. The scope of intervention included (1) alleviate problems of aches and malnutrition resulting from abdominal dropsy in the terminal stage of cancer; (2) provide psychological and spiritual care in order to ease patient fears of death and encourage positive reflection on life; (3) position the family to perform a positive role to help the patient and his family obtain the best quality of life. The patient finally passed away in the company of his family in a dignified, peaceful and painless manner.