RESUMEN
The Golgi apparatus is an essential organelle constructed by the stacking of flattened vesicles, that is widely distributed in eukaryotic cells and is dynamically regulated during cell cycles. It is a central station which is responsible for collecting, processing, sorting, transporting, and secreting some important proteins/enzymes from the endoplasmic reticulum to intra- and extra-cellular destinations. Golgi-specific fluorescent probes provide powerful non-invasive tools for the real-time and in situ visualization of the temporal and spatial fluctuations of bioactive species. Over recent years, more and more Golgi-targeting probes have been developed, which are essential for the evaluation of diseases including cancer. However, when compared with systems that target other important organelles (e.g. lysosomes and mitochondria), Golgi-targeting strategies are still in their infancy, therefore it is important to develop more Golgi-targeting probes. This review systematically summarizes the currently reported Golgi-specific fluorescent probes, and highlights the design strategies, mechanisms, and biological uses of these probes, we have structured the review based on the different targeting groups. In addition, we highlight the future challenges and opportunities in the development of Golgi-specific imaging agents and therapeutic systems.
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Colorantes Fluorescentes , Aparato de Golgi , Aparato de Golgi/metabolismo , Colorantes Fluorescentes/química , Humanos , AnimalesRESUMEN
Chromosome segregation must be under strict regulation to maintain chromosome euploidy and stability. Cell Division Cycle 20 (CDC20) is an essential cell cycle regulator that promotes the metaphase-to-anaphase transition and functions in the spindle assembly checkpoint, a surveillance pathway that ensures the fidelity of chromosome segregation. Plant CDC20 genes are present in multiple copies, and whether CDC20s have the same functions in plants as in yeast and animals is unclear, given the potential for divergence or redundancy among the multiple copies. Here, we studied all three CDC20 genes in rice (Oryza sativa) and constructed two triple mutants by clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9-mediated genome editing to explore their roles in development. Knocking out all three CDC20 genes led to total sterility but did not affect vegetative development. Loss of the three CDC20 proteins did not alter mitotic division but severely disrupted meiosis as a result of asynchronous and unequal chromosome segregation, chromosome lagging, and premature separation of chromatids. Immunofluorescence of tubulin revealed malformed meiotic spindles in microsporocytes of the triple mutants. Furthermore, cytokinesis of meiosis I was absent or abnormal, and cytokinesis II was completely prevented in all mutant microsporocytes; thus, no tetrads or pollen formed in either cdc20 triple mutant. Finally, the subcellular structures and functions of the tapetum were disturbed by the lack of CDC20 proteins. These findings demonstrate that the three rice CDC20s play redundant roles but are indispensable for faithful meiotic chromosome segregation and cytokinesis, which are required for the production of fertile microspores.
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División Celular/genética , Segregación Cromosómica/genética , Citocinesis/genética , Meiosis/genética , Oryza/genética , Productos Agrícolas/genética , Regulación de la Expresión Génica de las Plantas , Genes de PlantasRESUMEN
Hypochlorite (ClO-) plays an important role in the human immune defense system, but high concentrations of ClO- in the endoplasmic reticulum (ER) damage cellular proteins, causing ER stress, cell death, and various diseases. Herein, we developed a simple hydrazone probe (1) featuring aggregation-induced ratiometric emission, which would quickly (within 20 s) and sensitively (detection limit of 15.4 µM) respond to ClO- in an almost pure aqueous solution via a fluorescent ratiometric output. Furthermore, the probe was employed to track the level of ClO- in the ER of HeLa cells and zebrafish.
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Retículo Endoplásmico , Colorantes Fluorescentes , Ácido Hipocloroso , Animales , Humanos , Retículo Endoplásmico/metabolismo , Colorantes Fluorescentes/química , Células HeLa , Ácido Hipocloroso/análisis , Ácido Hipocloroso/metabolismo , Pez Cebra/metabolismoRESUMEN
The quality control system for messenger RNA (mRNA) is fundamental for cellular activities in eukaryotes. To elucidate the molecular mechanism of 3'-phosphoinositide-dependent protein kinase1 (PDK1), a master regulator that is essential throughout eukaryotic growth and development, we employed a forward genetic approach to screen for suppressors of the loss-of-function T-DNA insertion double mutant pdk1.1 pdk1.2 in Arabidopsis thaliana. Notably, the severe growth attenuation of pdk1.1 pdk1.2 was rescued by sop21 (suppressor of pdk1.1 pdk1.2), which harbors a loss-of-function mutation in PELOTA1 (PEL1). PEL1 is a homolog of mammalian PELOTA and yeast (Saccharomyces cerevisiae) DOM34p, which each form a heterodimeric complex with the GTPase HBS1 (HSP70 SUBFAMILY B SUPPRESSOR1, also called SUPERKILLER PROTEIN7, SKI7), a protein that is responsible for ribosomal rescue and thereby assures the quality and fidelity of mRNA molecules during translation. Genetic analysis further revealed that a dysfunctional PEL1-HBS1 complex failed to degrade the T-DNA-disrupted PDK1 transcripts, which were truncated but functional, and thus rescued the growth and developmental defects of pdk1.1 pdk1.2. Our studies demonstrated the functionality of a homologous PELOTA-HBS1 complex and identified its essential regulatory role in plants, providing insights into the mechanism of mRNA quality control.
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Proteínas de Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN de Planta/genética , ARN de Planta/metabolismoRESUMEN
Many type B polycyclic polyprenylated acylphloroglucinols (PPAPs) bear a lavandulyl-derived substituent, and the configurational assignment of this side chain can be difficult and sometimes leads to erroneous conclusions. In this study, 21 PPAPs, including the new xanthochymusones A-I (1-9), have been isolated from the fruits of Garcinia xanthochymus and structurally characterized. The relative configuration of the C-30 stereocenter was assigned by a combination of chemical transformations, 1H-1H coupling constants, conformational analysis, and NOE experiments. The configurational assignment of compound 7 indicates that the relative configuration at C-30 of PPAPs is not always the same. The absolute configurations of the new compounds were assigned by ECD and X-ray diffraction data, as well as by biosynthetic considerations. Analysis of NMR data enabled the configurational revision of garcicowins C and D. All the isolated PPAPs were tested for antiproliferative activity against three human hepatocellular carcinoma cell lines, including Huh-7, Hep 3B, and HepG2. Compounds 5 and 6, 7-epi-isogarcinol (16), and coccinone C (17) exhibited moderate antiproliferative activity. Compounds 6 and 16 induced apoptosis and inhibited cell migration in Huh-7 cells, probably through downregulating the STAT3 signaling pathway. This study provides effective methods for configurational assignments of type B PPAPs.
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Garcinia , Hypericum , Humanos , Garcinia/química , Frutas/química , Floroglucinol/farmacología , Floroglucinol/química , Conformación Molecular , Espectroscopía de Resonancia Magnética , Estructura Molecular , Hypericum/químicaRESUMEN
Abnormal increases in glucagon (GCG) are the primary cause of type II diabetes mellitus. When GCG interacts with a glucagon receptor (GCGR), GCG can increase the blood glucose level. In this paper, a compound that could interfere with the binding of GCG and GCGR to inhibit the increase of blood glucose was investigated. First, molecular docking was used to conduct preliminary screening of compounds whose active components could combine with GCGR by AutoDock Vina. The binding of the receptor-ligand complex was analyzed by PyMOL. Results showed that dauricine could tightly bind to the receptor pocket. Second, the plasmid pcDNA3.1(+)-GCGR containing the target gene was transfected into HEK293 cells for expression, which was the cell model established to screen GCGR antagonist. Dauricine, the lead compound of glucagon receptor antagonist (GRA), was screened using the GRA screening model in vitro. Finally, using [Des-His1, Glu9]-Glucagon amide as the positive control, flow cytometry was used to express the antagonistic effect of the compound. Consequently, dauricine can antagonize the GCGR.
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Diabetes Mellitus Tipo 2 , Receptores de Glucagón , Bencilisoquinolinas , Glucemia/metabolismo , Glucagón/metabolismo , Glucagón/farmacología , Células HEK293 , Humanos , Simulación del Acoplamiento Molecular , Receptores de Glucagón/genética , Receptores de Glucagón/metabolismo , TetrahidroisoquinolinasRESUMEN
Termites are serious pests in agriculture and forestry, causing significant economic losses to property and the construction industry. However, only a few entomopathogenic fungi attack termites that are dominant members of most terrestrial biomes. This study contributes to the taxonomic knowledge of entomopathogenic fungi with the description of a new pathogen on termites collected from the Pu Luong Nature Reserve in Vietnam. The new termite pathogen, Ophiocordyceps puluongensis, is introduced on the basis of morphological and multigene phylogenetic evidence. Based on the combined dataset of five genes including the nuclear ribosomal small and large subunits (nrSSU and nrLSU), the elongation factor 1α (tef-1α), and the largest and the second largest subunits of RNA polymerase II (rpb1 and rpb2), phylogenetic analyses were performed by Maximum Likelihood and Bayesian inference methods to determine the phylogenetic position of O. puluongensis. Three samples of O. puluongensis are clustered in the Hirsutella thompsonii subclade of Hirsutella lineages in Ophiocordyceps, and clustered together with O. asiatica to form a separate clade from other Ophiocordyceps species. Morphologically, O. puluongensis differs from O. asiatica by its smaller and shorter perithecia, asci and ascospores, pink to reddish-orange stipes of stromata, as well as smaller fusiform or citriform conidia. The distinctiveness of this termite pathogen is strongly supported by both molecular phylogeny and morphology. The entomopathogenic fungus O. puluongensis could have the potential to be used as bioinsecticides to control termites.
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Hypocreales , Isópteros , Animales , Teorema de Bayes , Hypocreales/genética , Filogenia , Esporas Fúngicas/genética , VietnamRESUMEN
This work developed a sensitive electrochemiluminescence (ECL) biosensor based on a cyclometalated iridium(III) complex ((bt)2Irbza), which was synthesized for the first time. Annihilation, reductive-oxidative, and oxidative-reductive ECL behaviors of (bt)2Irbza were investigated, respectively. The oxidative-reductive ECL intensity was the strongest compared with the other two, which showed 16.7 times relative ECL efficiency compared with commercial [Ru(bpy)3]2+ under the same experimental conditions. Therefore, an ECL biosensing system with (bt)2Irbza as the anodic luminophore was established for miRNA detection based on a closed bipolar electrode (BPE). Combined with both steric hindrance and catalytic effects induced by hemin/G-quadruplex in the cathodic reservoir of BPE that changed the Faraday current of the cathode and thus mediated the ECL intensity of (bt)2Irbza in the anode of BPE, the ECL sensor stated an ultrahigh sensitivity for microRNA (miRNA-122) analysis with a detection limit of 82 aM.
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Técnicas Biosensibles , Complejos de Coordinación/química , Técnicas Electroquímicas , Iridio/química , Mediciones Luminiscentes , MicroARNs/análisis , Complejos de Coordinación/síntesis química , Electrodos , Humanos , Estructura MolecularRESUMEN
Although clinical data suggest remarkable promise for targeting programmed cell death protein-1 (PD-1) and ligand (PD-L1) signaling in non-small-cell lung cancer (NSCLC), it is still largely undetermined which subtype of patients will be responsive to checkpoint blockade. In the present study, we explored whether PD-L1 was regulated by mutant Kirsten rat sarcoma viral oncogene homolog (KRAS), which is frequently mutated in NSCLC and results in poor prognosis and low survival rates. We verified that PD-L1 levels were dramatically increased in KRAS mutant cell lines, particularly in NCI-H441 cells with KRAS G12V mutation. Overexpression of KRAS G12V remarkably elevated PD-L1 messenger RNA and protein levels, while suppression of KRAS G12V led to decreased PD-L1 levels in NCI-H441 cells. Consistently, higher levels of PD-L1 were observed in KRAS-mutated tissues as well as tumor tissues-derived CD4+ and CD8+ T cells using a tumor xenograft in B-NDG mice. Mechanically, both in vitro and in vivo assays found that KRAS G12V upregulated PD-L1 via regulating the progression of epithelial-to-mesenchymal transition (EMT). Moreover, pembrolizumab activated the antitumor activity and decreased tumor growth with KRAS G12V mutated NSCLC. This study demonstrates that KRAS G12V mutation could induce PD-L1 expression and promote immune escape via transforming growth factor-ß/EMT signaling pathway in KRAS-mutant NSCLC, providing a potential therapeutic approach for NSCLC harboring KRAS mutations.
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Anticuerpos Monoclonales Humanizados/farmacología , Antígeno B7-H1/metabolismo , Carcinoma de Pulmón de Células no Pequeñas , Proteínas Proto-Oncogénicas p21(ras)/genética , Animales , Biomarcadores de Tumor/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Línea Celular Tumoral , Transición Epitelial-Mesenquimal , Humanos , Ratones , Ratones Endogámicos NOD , Ratones SCIDRESUMEN
Cordyceps militaris is a traditional Chinese medicinal food that is challenging to quality maintaining while mass cultivation. Many studies have found that abundant microbes inhabit Ophiocordyceps sinensis and perform important functions for their host. In this study, our objective was to reveal the microbial communities that inhabit C. militaris and analyze their potential functions. High-throughput sequencing of 16S rRNA and ITS genes was used to compare the diversity and composition of the bacterial and fungal communities associated with naturally occurring C. militaris collected from Yunnan Province, southwestern China. The diversity and richness of the microbial communities and the number of function genes of the bacteria were significantly higher in the habitat soil than in the fruiting body. The sclerotia and stromata samples shared the same microbiota and functions. The main bacterial phyla were Proteobacteria, Acidobacteria, Bacteroidetes and Actinobacteria, and Ascomycota was the main fungal phylum. The growth-promoting bacteria Herbaspirillum and the plant probiotic Phyllobacterium, which may enhance C. militaris quality and facilitate its cultivation, were detected in the fruiting body samples. Genes related to metabolism were more abundant in the soil bacteria, while membrane transport genes were more abundant in the endophytic bacteria of C. militaris. Our study is the first to reveal the unexpectedly high diversity of the microbial communities and the bacterial functions inhabiting the natural C. militaris using high-throughput sequencing, and our results provide insights into mining the functions of microorganisms in the development and quality of C. militaris.
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Cordyceps , Microbiota , China , Cordyceps/genética , ARN Ribosómico 16S/genética , Suelo , Microbiología del SueloRESUMEN
A total of 15 novel-substituted 3-(benzylsulfanyl)-1H-1,2,4-triazol-5-ylamine and 10 novel-substituted 3-benzylmercapto-1,2,4-triazol derivatives were synthesized based on the natural product phenazine-1-carboxylic acid (PCA). Their structures were confirmed by 1H-NMR, 13C-NMR, HRMS, and X-ray. Most substituted 3-benzylmercapto-1,2,4-triazol derivatives displayed very strong fungicidal activity against one or multiple plant pathogens in vitro and in vivo. Compounds 8b, 8h, and 8i showed a broad spectrum of fungicidal activity. Further field experiments indicated that compounds 8b, 8c, and 8h displayed better efficacy against rice blast (Pyricularia oryzae) than PCA. These data demonstrate that compounds 8b, 8c, and 8h are promising fungicidal candidates, deserving further studies.[Formula: see text].
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Fungicidas Industriales , Fenazinas , Ascomicetos , Fungicidas Industriales/farmacología , Estructura Molecular , Fenazinas/farmacología , Relación Estructura-Actividad , Triazoles/farmacologíaRESUMEN
Root hair development is crucial for phosphate absorption, but how phosphorus deficiency affects root hair initiation and elongation remains unclear. We demonstrated the roles of auxin efflux carrier PIN-FORMED2 (PIN2) and phospholipase D (PLD)-derived phosphatidic acid (PA), a key signaling molecule, in promoting root hair development in Arabidopsis thaliana under a low phosphate (LP) condition. Root hair elongation under LP conditions was greatly suppressed in pin2 mutant or under treatment with a PLDζ2-specific inhibitor, revealing that PIN2 and polar auxin transport and PLDζ2-PA are crucial in LP responses. PIN2 was accumulated and degraded in the vacuole under a normal phosphate (NP) condition, whereas its vacuolar accumulation was suppressed under the LP or NP plus PA conditions. Vacuolar accumulation of PIN2 was increased in pldζ2 mutants under LP conditions. Increased or decreased PIN2 vacuolar accumulation is not observed in sorting nexin1 (snx1) mutant, indicating that vacuolar accumulation of PIN2 is mediated by SNX1 and the relevant trafficking process. PA binds to SNX1 and promotes its accumulation at the plasma membrane, especially under LP conditions, and hence promotes root hair development by suppressing the vacuolar degradation of PIN2. We uncovered a link between PLD-derived PA and SNX1-dependent vacuolar degradation of PIN2 in regulating root hair development under phosphorus deficiency.
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Proteínas de Arabidopsis , Fosfolipasa D , Proteínas de Arabidopsis/genética , Ácidos Indolacéticos , Ácidos Fosfatidicos , Fosfolipasa D/genética , Fósforo , Raíces de Plantas/fisiología , VacuolasRESUMEN
The Arabidopsis (Arabidopsis thaliana) root epidermis is a simple model for investigating cell fate specification and pattern formation. In addition to regulatory networks consisting of transcription factors, histone deacetylases are also involved in the formation of cellular patterns. Here, we report thatHistone Deacetylase19 (HDA19) affects the root epidermal cellular pattern through regulation of cortical cell fate by interacting with SCARECROW (SCR). HDA19 binds to the DNA sequence upstream of SCR, as well as to those of several of SCR's target genes, and regulates their expression. Mutant lines of several SCR target genes show impaired patterns of epidermal differentiation and cortical cell division, similar to that of hda19 This work presents HDA19 and SCR as two further players in the regulation of cortical and epidermal cell specification and describes an additional function for SCR.
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Proteínas de Arabidopsis/metabolismo , Arabidopsis/citología , Histona Desacetilasas/metabolismo , Raíces de Plantas/citología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Diferenciación Celular , Regulación de la Expresión Génica de las Plantas , Histona Desacetilasas/genética , Células Vegetales , Epidermis de la Planta/citología , Raíces de Plantas/metabolismo , Plantas Modificadas GenéticamenteRESUMEN
This paper describes a bidirectional electrochemiluminescence (ECL) biosensor for the detection of microRNA-141 (miRNA-141) with aM level detection limit and two-segment 8 orders of magnitude linear range. Hemin/G-quadruplex DNAzyme was assembled on carbon nitride nanosheets and Au nanoparticles modified electrode. The ECL of carbon nitride nanosheets could be enhanced by the right amount of H2O2 (no more than 10 mM) and then inhibited by excessive H2O2 when 0.1 M K2S2O8 acted as coreactant. In the presence of excessive H2O2 (20 mM), a recovery of ECL intensity was obtained due to the catalytic reduction of H2O2 caused by hemin/G-quadruplex DNAzyme at lower target concentrations, and then an ECL decrease occurred mainly by biocatalytic precipitation (BCP)-induced charge transfer resistance on the electrode surface at higher target concentrations. Therefore, based on the change of the ECL intensity caused by the catalytic reduction of H2O2 and BCP, a highly sensitive bidirectional miRNA sensor with ultralow detection limit of 7.9 aM and wide linear range from 10-17 to 10-9 M was obtained. This work could attract more attention on the study of multiple mechanisms and also provides a more sensitive and precise method for the analysis of nucleic acids.
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Técnicas Biosensibles , Técnicas Electroquímicas , Luminiscencia , MicroARNs/análisis , HumanosRESUMEN
This study aimed at elucidating regulatory components behind floral organ identity determination and tissue development. It remains unclear how organ identity proteins facilitate development of organ primordia into tissues with a determined identity, even though it has long been accepted that floral organ identity is genetically determined by interaction of identity genes according to the ABC model. Using the chromatin immunoprecipitation sequencing technique, we identified OsTGA10, encoding a bZIP transcription factor, as a target of the MADS box protein OsMADS8, which is annotated as an E-class organ identity protein. We characterized the function of OsTGA10 using genetic and molecular analyses. OsTGA10 was preferentially expressed during stamen development, and mutation of OsTGA10 resulted in male sterility. OsTGA10 was required for tapetum development and functioned by interacting with known tapetum genes. In addition, in ostga10 stamens, the hallmark cell wall thickening of the endothecium was defective. Our findings suggest that OsTGA10 plays a mediator role between organ identity determination and tapetum development in rice stamen development, between tapetum development and microspore development, and between various regulatory components required for tapetum development. Furthermore, the defective endothecium in ostga10 implies that cell wall thickening of endothecium is dependent on tapetum development.
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Flores/crecimiento & desarrollo , Flores/metabolismo , Proteínas de Dominio MADS/metabolismo , Oryza/crecimiento & desarrollo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Pared Celular/metabolismo , ADN de Plantas/metabolismo , Epistasis Genética , Flores/citología , Flores/ultraestructura , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Oryza/genética , Fenotipo , Homología de Secuencia de AminoácidoRESUMEN
Germ cells are indispensable carriers of genetic information from one generation to the next. In contrast to the well-understood process in animals, information on the mechanism of germ cell initiation in plants is very limited. SPOROCYTELESS/NOZZLE was previously identified as an essential regulator of diploid germ cell (archesporial cell) differentiation in the stamens and ovules of Arabidopsis (Arabidopsis thaliana). Although SPOROCYTELESS (SPL) transcription is activated by the floral organ identity regulator AGAMOUS and epigenetically regulated by SET DOMAIN GROUP2, little is known about the regulation of the SPL protein. Here, we report that the protein kinases MPK3 and MPK6 can both interact with SPL in vitro and in vivo and can phosphorylate the SPL protein in vitro. In addition, phosphorylation of the SPL protein by MPK3/6 is required for SPL function in the Arabidopsis anther, as measured by its effect on archesporial cell differentiation. We further demonstrate that phosphorylation enhances SPL protein stability. This work not only uncovers the importance of SPL phosphorylation for its regulatory role in Arabidopsis anther development, but also supports the hypothesis that the regulation of precise spatiotemporal patterning of germ cell initiation and that differentiation is achieved progressively through multiple levels of regulation, including transcriptional and posttranslational modification.
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Proteínas de Arabidopsis/metabolismo , Flores/metabolismo , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Represoras/metabolismo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Diferenciación Celular/genética , Flores/genética , Flores/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Células Germinativas de las Plantas/citología , Células Germinativas de las Plantas/metabolismo , Inmunohistoquímica , Microscopía Fluorescente , Quinasas de Proteína Quinasa Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/genética , Mutación , Proteínas Nucleares/genética , Fosforilación , Plantas Modificadas Genéticamente , Unión Proteica , Estabilidad Proteica , Proteínas Represoras/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Purpose To evaluate the safety and efficacy of microwave ablation (MWA) in patients with end-stage renal disease and secondary hyperparathyroidism. Materials and Methods The study protocol was approved by the human ethics review committee. Between March 1, 2014, and June 30, 2015, 51 patients (25 men, 26 women; mean age ± standard deviation, 53.1 years ± 12.9) were enrolled. All patients had at least one enlarged parathyroid gland and secondary symptomatic hyperparathyroidism, which was treated with ultrasonographically (US) guided MWA. The levels of intact parathyroid hormone, serum calcium, phosphorus, and alkaline phosphatase were compared before and after MWA. Paired-sample t tests and paired-sample Wilcoxon signed-rank tests were used to compare treatment outcomes before and after MWA. Results Complete ablation was achieved in all 96 glands in 51 of 120 patients with severe secondary hyperparathyroidism. The mean follow-up time was 11.1 months ± 3.3. The maximum diameter of the glands ranged from 0.5 cm to 4.8 cm (mean, 1.5 cm ± 0.6). The ablation time for each gland was 216.1 seconds ± 130.1. The mean serum intact parathyroid hormone, calcium, and phosphorus levels after MWA (400 pg/mL [400 ng/L; range, 151.3-629.0 ng/L], 2.33 mmol/L ± 0.23, and 1.54 mmol/L ± 0.43, respectively) were significantly lower than those before MWA (1203 pg/mL [1203 ng/L; range, 854.7-1694.5 ng/L], 2.53 mmol/L ± 0.24, and 1.97 mmol/L ± 0.50, respectively; P < .01), while the alkaline phosphatase levels did not change with MWA (P > .05). Ipsilateral recurrent laryngeal nerve injury was seen in one patient (2%). A hematoma developed during one procedure in one patient (2%) and was treated successfully with injection of thrombin. Conclusion US-guided MWA is safe and effective for destroying parathyroid gland tissue in patients with end-stage renal disease and severe secondary hyperparathyroidism. Further experience with the technique is clearly necessary. © RSNA, 2016.
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Ablación por Catéter/métodos , Hiperparatiroidismo Secundario/etiología , Hiperparatiroidismo Secundario/cirugía , Fallo Renal Crónico/complicaciones , Microondas , Glándulas Paratiroides/cirugía , Ultrasonografía Intervencional , Femenino , Humanos , Hiperplasia , Masculino , Persona de Mediana Edad , Proyectos Piloto , Resultado del TratamientoRESUMEN
Cellular patterning in the Arabidopsis (Arabidopsis thaliana) root epidermis is dependent on positional information, the transmission of which involves histone acetylation. Here, we report that HISTONE DEACETYLASE6 (HDA6) has significant effects on this cellular patterning. Mutation of HDA6 led to ectopic hair cells in the nonhair positions of root epidermis in Arabidopsis, based on an analysis of paraffin sections stained with Toluidine Blue. While HDA6 was present throughout the root tip, epidermis-specific complementation with HDA6 could rescue the hda6 phenotype. Both transcript levels and expression patterns of ENHANCER OF TRIPTYCHON AND CAPRICE1 (ETC1) and GLABRA2 (GL2) in the root tip were affected in hda6. Consistent with these changes in expression, HDA6 directly bound to the promoter regions of ETC1 and GL2, and acetylation of histone H3 on these promoter regions and acetylation of histone H4 on the ETC1 promoter region was increased in the hda6 mutant. Taken together, these results indicate that HDA6 affects the cellular patterning of Arabidopsis root epidermis through altering the histone acetylation status of ETC1 and GL2 promoters and thereby affects the expression of these two components of the core transcription factor network determining epidermal cell fates. Our findings thus provide new insights into the role of histone acetylation in root epidermis cell patterning.
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Proteínas de Arabidopsis/genética , Proteínas de Unión al ADN/genética , Histona Desacetilasas/genética , Proteínas de Homeodominio/genética , Mutación , Epidermis de la Planta/genética , Raíces de Plantas/genética , Acetilación , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Unión al ADN/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Histona Desacetilasas/metabolismo , Proteínas de Homeodominio/metabolismo , Microscopía Confocal , Epidermis de la Planta/metabolismo , Raíces de Plantas/metabolismo , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Plantones/genética , Plantones/metabolismoRESUMEN
The differentiation of hair (H) and non-hair (N) cells in the Arabidopsis thaliana root epidermis is dependent on positional relationships with underlying cortical cells. We previously found that histone acetylation relays positional information and that a mutant altered in the histone deacetylase gene family member HISTONE DEACETYLASE 18 (HDA18) exhibits altered H and N epidermal cell patterning. Here, we report that HDA18 has in vitro histone deacetylase activity and that both mutation and overexpression of HDA18 led to cells at the N position having H fate. The HDA18 protein physically interacted with histones related to a specific group of kinase genes, which are demonstrated in this study to be components of a positional information relay system. Both down- and upregulation of HDA18 increased transcription of the targeted kinase genes. Interestingly, the acetylation levels of histone 3 lysine 9 (H3K9), histone 3 lysine 14 (H3K14) and histone 3 lysine 18 (H3K18) at the kinase genes were differentially affected by down- or upregulation of HDA18, which explains why the transcription levels of the four HDA18-target kinase genes increased in all lines with altered HDA18 expression. Our results reveal the surprisingly complex mechanism by which HDA18 affects cellular patterning in Arabidopsis root epidermis.