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Images document scientific discoveries and are prevalent in modern biomedical research. Microscopy imaging in particular is currently undergoing rapid technological advancements. However, for scientists wishing to publish obtained images and image-analysis results, there are currently no unified guidelines for best practices. Consequently, microscopy images and image data in publications may be unclear or difficult to interpret. Here, we present community-developed checklists for preparing light microscopy images and describing image analyses for publications. These checklists offer authors, readers and publishers key recommendations for image formatting and annotation, color selection, data availability and reporting image-analysis workflows. The goal of our guidelines is to increase the clarity and reproducibility of image figures and thereby to heighten the quality and explanatory power of microscopy data.
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Lista de Checagem , Editoração , Reprodutibilidade dos Testes , Processamento de Imagem Assistida por Computador , MicroscopiaRESUMO
Flowering is critical for sexual reproduction and fruit production. Several pear (Pyrus sp.) varieties produce few flower buds, but the underlying mechanisms are unknown. The circadian clock regulator EARLY FLOWERING3 (ELF3) serves as a scaffold protein in the evening complex that controls flowering. Here, we report that the absence of a 58-bp sequence in the 2nd intron of PbELF3 is genetically associated with the production of fewer flower buds in pear. From rapid amplification of cDNA ends sequencing results, we identified a short, previously unknown transcript from the PbELF3 locus, which we termed PbELF3ß, whose transcript level was significantly lower in pear cultivars that lacked the 58-bp region. The heterologous expression of PbELF3ß in Arabidopsis (Arabidopsis thaliana) accelerated flowering, whereas the heterologous expression of the full-length transcript PbELF3α caused late flowering. Notably, ELF3ß was functionally conserved in other plants. Deletion of the 2nd intron reduced AtELF3ß expression and caused delayed flowering time in Arabidopsis. AtELF3ß physically interacted with AtELF3α, disrupting the formation of the evening complex and consequently releasing its repression of flower induction genes such as GIGANTEA (GI). AtELF3ß had no effect in the absence of AtELF3α, supporting the idea that AtELF3ß promotes flower induction by blocking AtELF3α function. Our findings show that alternative promoter usage at the ELF3 locus allows plants to fine-tune flower induction.
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Proteínas de Arabidopsis , Arabidopsis , Relógios Circadianos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Relógios Circadianos/fisiologia , Plantas/metabolismo , Flores/metabolismoRESUMO
Self-incompatibility (SI) is a widespread genetically determined system in flowering plants that prevents self-fertilization to promote gene flow and limit inbreeding. S-RNase-based SI is characterized by the arrest of pollen tube growth through the pistil. Arrested pollen tubes show disrupted polarized growth and swollen tips, but the underlying molecular mechanism is largely unknown. Here, we demonstrate that the swelling at the tips of incompatible pollen tubes in pear (Pyrus bretschneideri [Pbr]) is mediated by the SI-induced acetylation of the soluble inorganic pyrophosphatase (PPA) PbrPPA5. Acetylation at Lys-42 of PbrPPA5 by the acetyltransferase GCN5-related N-acetyltransferase 1 (GNAT1) drives accumulation of PbrPPA5 in the nucleus, where it binds to the transcription factor PbrbZIP77, forming a transcriptional repression complex that inhibits the expression of the pectin methylesterase (PME) gene PbrPME44. The function of PbrPPA5 as a transcriptional repressor does not require its PPA activity. Downregulating PbrPME44 resulted in increased levels of methyl-esterified pectins in growing pollen tubes, leading to swelling at their tips. These observations suggest a mechanism for PbrPPA5-driven swelling at the tips of pollen tubes during the SI response. The targets of PbrPPA5 include genes encoding cell wall-modifying enzymes, which are essential for building a continuous sustainable mechanical structure for pollen tube growth.
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Tubo Polínico , Pyrus , Ribonucleases/metabolismo , Pirofosfatase Inorgânica/genética , Pirofosfatase Inorgânica/metabolismo , Acetilação , Pyrus/metabolismoRESUMO
The high prevalence of breast cancer is a global health concern, compounded by the lack of safe or effective treatments for its advanced stages. These facts urge the development of novel treatment strategies. Annexin A5 (ANXA5) is a natural human protein that binds with high specificity to phosphatidylserine, a phospholipid tightly maintained in the inner leaflet of the cell membrane on most healthy cells but externalized in tumor cells and the tumor vasculature. Here, we have developed a targeted photosensitizer for photothermal therapy (PTT) of solid tumors through the functionalization of single-walled carbon nanotubes (SWCNTs) to ANXA5-the SWCNT-ANXA5 conjugate. The ablation of tumors through the SWCNT-ANXA5-mediated PTT synergizes with checkpoint inhibition, creating a systemic anticancer immune response. In vitro ablation of cells incubated with the conjugate promoted cell death in a dose-dependent and targeted manner. This treatment strategy was tested in vivo with the orthotopic EMT6 breast tumor model in female balb/cJ mice. Enhanced therapeutic effects were achieved by using intratumoral injection of the conjugate and treating tumors at a lower PTT temperature (45°C). Intratumoral injection prevented the accumulation of the SWCNTs in major clearance organs. When combined with checkpoint inhibition of anti-programmed cell death protein-1, SWCNT-ANXA5-mediated PTT increased survival and 80% of the mice survived for 100 days. Evidence of immune system activation by flow cytometry of splenic cells strengthens the hypothesis of an abscopal effect as a mechanism of prolonged survival. SIGNIFICANCE STATEMENT: This study demonstrated a relatively high survival rate (80% at 100 days) of mice with aggressive breast cancer when treated with photothermal therapy using the SWCNT-ANXA5 conjugate injected intratumorally and combined with immune stimulation using the anti-programmed cell death protein-1 checkpoint inhibitor. Photothermal therapy was accomplished by maintaining the tumor temperature at a relatively low level of 45°C and avoiding accumulation of the nanotubes in the clearance organs by using intratumoral administration.
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Neoplasias da Mama , Camundongos Endogâmicos BALB C , Nanotubos de Carbono , Terapia Fototérmica , Nanotubos de Carbono/química , Animais , Feminino , Camundongos , Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Neoplasias da Mama/imunologia , Terapia Fototérmica/métodos , Linhagem Celular Tumoral , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Receptor de Morte Celular Programada 1/imunologia , Humanos , Metástase Neoplásica , Imunoterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Fototerapia/métodosRESUMO
DNA methylation plays an important role in regulating tomato (Solanum lycopersicum) fruit ripening. Although SlDML2, a DNA demethylase (DML) gene, is critically involved in tomato fruit ripening, little is known about genes that regulate its expression. Using yeast one-hybrid screening, we identified a High Mobility Group A protein, named SlHMGA3, and demonstrated its binding activity to the AT-rich region of the SlDML2 promoter. We produced slhmga3 tomato mutants using CRISPR/Cas9 and observed that slhmga3 fruit reached the breaker stage much later than fruit from the wild-type. We further demonstrated that at the initiation stage of fruit ripening, the increased expression of SlDML2 and ethylene biosynthetic and signaling genes was significantly delayed in slhmga3 fruit, along with delays in ethylene production and demethylation and activation of ripening-associated transcription factor genes. Our results demonstrate that SlHMGA3 plays a role in enhancing SlDML2 expression, and its effects on tomato fruit ripening are largely through DNA demethylation of ripening-associated transcription factor genes.
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Solanum lycopersicum , DNA/metabolismo , Etilenos/metabolismo , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Extracellular polymeric substances (EPS) are extracellular metabolites of microorganisms, highly associated with microbial function, adaptation, and growth. The main compounds in EPS have been revealed to be proteins, polysaccharides, nucleic acids, humic substances, lipids, etc. EPS are not only biomass, but also a biogenic material. EPS have high specific surface, abundant functional groups, and excellent degradability. In addition, they are more extensible to the environment than the microbial cells themselves, which exhibits their huge advantages. Therefore, they have been applied in many fields, such as the environment, ecosystem, basic commodities, and medicine. However, the functions of EPS highly depend on the suitable extraction process, as different extraction methods have different effects on their composition, structure, and function. There are many types of EPS extraction methods, in which physical and chemical methods have been widely utilized. This review summarizes the extraction methods and applications of EPS. In addition, it considers some important gaps in current knowledge, and indicates perspectives of EPS for their future study.
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Matriz Extracelular de Substâncias Poliméricas , Fungos , Fungos/metabolismo , Fungos/química , Fungos/genética , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Matriz Extracelular de Substâncias Poliméricas/química , Polímeros/metabolismo , Polímeros/químicaRESUMO
Many fungi are able to produce extracellular polymeric substances (EPS) for environmental, food, and industrial applications. This study evaluated the extraction (in vivo) of EPS from Rhodotorula mucilaginosa, a typical yeast with abundant EPS. Three extracting methods were set, i.e., heating, addition of NaCl during heating, and cation exchange resin (CER). The abundance of extracted proteins and polysaccharides showed evident contrasts (elevated to ~ 600 and 1700 mg/L, respectively) after heating at 70 °C in water. Although the higher temperature will increase the extracted abundance of EPS, the leakage of DNA would be enhanced due to cell rupture. The addition of NaCl further promoted the efficiency of extraction, either for proteins (from ~ 550 to ~ 650 mg/L) or polysaccharides (from ~ 1700 to ~ 2010 mg/L). Moreover, the biochemical results showed that the extracted abundance of EPS via heating was dramatically higher than that via CER. Additionally, DNA leakage in the CER treatment (2.0 g/g DW) was significantly higher (up to > 6 mg/L) than that under heating at 70 °C (< 2 mg/L). Furthermore, the three-dimensional excitation-emission matrix spectra showed two characteristic peaks of emission/excitation wavelength at 280/300 and 280/350, suggesting the relative high diversity of organic matters in EPS after heating treatments. Finally, a fluctuation of polysaccharide abundance in EPS at 500-1500 mg/L Pb2+ level was elucidated by the extraction based on heating treatment. This study hence confirmed that the heating method might be recommended for extraction of EPS from fungi in vivo KEY POINTS: ⢠3D-EEM results indicated that heating could extract more EPS compared with CER. ⢠Heating treatments showed lower DNA leakage from fungi than CER treatments. ⢠Addition of NaCl promoted the detachment of EPS from fungal cells in vivo.
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Matriz Extracelular de Substâncias Poliméricas , Cloreto de Sódio , Polissacarídeos , Proteínas , Fungos , Esgotos/químicaRESUMO
It is of great significance to study the thermal radiation anomalies of earthquake swarms in the same area in terms of selecting abnormal characteristic determination parameters, optimizing and determining the processing model, and understanding the abnormal machine. In this paper, we investigated short-term and long-term thermal radiation anomalies induced by earthquake swarms in Iran and Pakistan between 2007 and 2016. The anomalies were extracted from infrared remote sensing black body temperature data from the China Geostationary Meteorological Satellites (FY-2C/2E/2F/2G) using the multiscale time-frequency relative power spectrum (MS T-FRPS) method. By analyzing and summarizing the thermal radiation anomalies of series earthquake groups with consistency law through a stable and reliable MS T-FRPS method, we first obtained the relationship between anomalies and ShakeMaps from USGS and proposed the anomaly regional indicator (ARI) to determine seismic anomalies and the magnitude decision factor (MDF) to determine seismic magnitude. In addition, we explored the following discussions: earthquake impact on regional thermal radiation background and the relationship between thermal anomalies and earthquake magnitude and the like. Future research directions using the MS T-FRPS method to characterize regional thermal radiation anomalies induced by strong earthquakes could help improve the accuracy of earthquake magnitude determination.
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Diabetes can cause vascular remodelling and is associated with worse outcome after ischaemic stroke. Pioglitazone is a commonly used anti-diabetic agent. However, it is not known whether pioglitazone use before ischaemia could reduce brain ischaemic injury. Pioglitazone was administered to 5-week-old db+ or db/db mice. Cerebral vascular remodelling was examined at the age of 9 weeks. Expression of peroxisome proliferator-activated receptor-γ (PPARγ), p-PPARγ (S112 and S273), nucleotide-binding domain (NOD)-like receptor protein 3 (Nlrp3), interleukin-1ß (IL-1ß) and tumour necrosis factor-α (TNF-α) was evaluated in the somatosensory cortex of mice. Neurological outcome was evaluated 24 h after brain ischaemia. Results showed that early pioglitazone treatment provided a long-lasting effect of euglycaemia but enhanced hyperlipidaemia in the db/db mice. Diabetic mice exhibited increased vascular tortuosity, narrower middle cerebral artery (MCA) width and IgG leakage in the brain. These changes were blocked by early pioglitazone treatment. In diabetic animals, PPARγ expression was reduced, and p-PPARγ at S273 but not S112, Nlrp3, IL-1ß and TNF-α were increased in the somatosensory cortex. PPARγ decrease and Nlrp3 increase were mainly in the neurons of the diabetic brain, which was reversed by early pioglitazone treatment. Pioglitazone attenuated the aggravated neurological outcome after stroke in diabetic mice. But this protective effect was abolished through restoring cerebral inflammation by intracerebroventricular administration of IL-1ß and TNF-α in pioglitazone-treated diabetic mice before MCAO. In summary, early pioglitazone treatment attenuates cerebral vascular remodelling and ischaemic brain injury possibly via blocking chronic neuroinflammation in the db/db mice.
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Isquemia Encefálica , Diabetes Mellitus Experimental , AVC Isquêmico , Acidente Vascular Cerebral , Animais , Isquemia Encefálica/tratamento farmacológico , Diabetes Mellitus Experimental/complicações , Inflamação/complicações , Inflamação/tratamento farmacológico , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR , PPAR gama/metabolismo , Pioglitazona/farmacologia , Pioglitazona/uso terapêutico , Acidente Vascular Cerebral/complicações , Fator de Necrose Tumoral alfa , Remodelação VascularRESUMO
Rhodotorula mucilaginosa resists heavy metal (HM) stress because of its abundant extracellular polymeric substances and functional vesicles. In this study, we provided new insights into its survival strategies at both biochemical and genetic levels. After lead exposure, carotenoid biosynthesis was initiated within 24 h incubation and then increased to the maximum after 96 h of incubation. Raman analysis confirmed that carotenoids (primarily ß-carotene) were the major identifiable chemical substances on the cell surface. Moreover, the increased carotenoid production was accompanied by a rising budding rate, ~40% higher than that in the cultures without Pb. During the 96 h of incubation, the driving force for Pb accumulation was assigned to this elevated budding rate. After 96 h, biosorption was primarily attributed to the enhanced antioxidant ability of the single cells during carotenoid production. Furthermore, the yeast budding cells demonstrated an evidently heterogeneous biosorption of Pb, i.e., the rejuvenated daughters had a relatively lower Pb level than the mother cells. This resulted in the protection of the buds from Pb stress. After investigating phosphorus uptake and the RNA sequencing data, we finally confirmed two tightly correlated pathways that resist HM stress, i.e., biochemical (carotenoid production) and reproductive (healthy buds) pathways.
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Chumbo , Rhodotorula , Carotenoides/metabolismo , Rhodotorula/genética , Rhodotorula/metabolismo , beta CarotenoRESUMO
The vital function of mounting long noncoding RNAs (lncRNAs) in prostate cancer (PCa) has been illustrated in increasing reports. However, the roles of many lncRNAs in PCa have not been deciphered. A total of 62 pairs of PCa and adjacent normal tissue samples were provided by PCa patients undergoing surgery. Extensive assays were conducted in this study to investigate the role of FOXP4 antisense RNA 1 (FOXP4-AS1) in PCa tumorigenesis. This study elucidated that FOXP4-AS1 expression was elevated in PCa tissue samples and cell lines. Loss-of-function experiments revealed that depleted FOXP4-AS1 inhibited PCa cell proliferation in vitro and retarded tumor growth in vivo. Mechanically, FOXP4-AS1 functioned as a competing endogenous RNA (ceRNA) of miR-3130-3p, releasing SP4 from the inhibitory effect of miR-3130-3p. Rescue assays validated that FOXP4-AS1 modulated PCa progression via SP4. Interestingly, SP4 is known as a transcription factor and was predicted to bind with the promoter region of FOXP4-AS1. This current research confirmed that SP4 activated the transcription activity of FOXP4-AS1 and thus positively regulated its expression. To conclude, we discovered that FOXP4-AS1, miR-3130-3p, and SP4 constitute a feedback loop and contribute to PCa tumorigenesis, providing a new valuable diagnosis and therapeutic strategy for PCa.
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MicroRNAs , Neoplasias da Próstata , RNA Longo não Codificante , Humanos , Masculino , Carcinogênese/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Retroalimentação , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , MicroRNAs/metabolismo , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismoRESUMO
Ischemic conditioning and exercise have been suggested for protecting against brain ischemia-reperfusion injury. However, the endogenous protective mechanisms stimulated by these interventions remain unclear. Here, in a comprehensive translational study, we investigated the protective role of extracellular vesicles (EVs) released after remote ischemic conditioning (RIC), blood flow restricted resistance exercise (BFRRE), or high-load resistance exercise (HLRE). Blood samples were collected from human participants before and at serial time points after intervention. RIC and BFRRE plasma EVs released early after stimulation improved viability of endothelial cells subjected to oxygen-glucose deprivation. Furthermore, post-RIC EVs accumulated in the ischemic area of a stroke mouse model, and a mean decrease in infarct volume was observed for post-RIC EVs, although not reaching statistical significance. Thus, circulating EVs induced by RIC and BFRRE can mediate protection, but the in vivo and translational effects of conditioned EVs require further experimental verification.
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Vesículas Extracelulares , Traumatismo por Reperfusão , Animais , Modelos Animais de Doenças , Células Endoteliais , Humanos , Isquemia , CamundongosRESUMO
MAIN CONCLUSION: The possible candidate expansin genes, which may be important for strawberry fruit softening, have been identified in the diploid woodland strawberry Fragaria vesca and the octoploid cultivated strawberry Fragaria × ananassa and their transcriptional regulation by histone modifications has been studied. Softening process greatly affects fruit texture and shelf life. Expansins (EXPs) are a group of structural proteins participating in cell wall loosening, which break the hydrogen bonding between cellulose microfibrils and hemicelluloses. However, our knowledge on how EXP genes are regulated in fruit ripening, especially in non-climacteric fleshy fruits, is limited. Here, we have identified the EXP genes in both the octoploid cultivated strawberry (Fragaria × ananassa) and one of its diploid progenitor species, woodland strawberry (Fragaria vesca). We found that EXP proteins in F. × ananassa were structurally more divergent than the ones in F. vesca. Transcriptome data suggested that FaEXP88, FaEXP114, FveEXP11 and FveEXP33 were the four candidate EXP genes more likely involved in fruit softening, whose transcript levels dramatically increased when firmness decreased during fruit maturation. Phylogenetic analyses showed that those candidate genes were closely clustered, indicating the presence of homoeolog expression dominance in the EXP gene family in strawberry. Moreover, we have performed chromatin immunoprecipitation (ChIP) experiments to investigate the distribution of histone modifications along the promoters and genic regions of the EXP genes in F. vesca. ChIP data revealed that the transcript levels of EXP genes were highly correlated with the enrichment of H3K9/K14 acetylation and H3K27 tri-methylation. Collectively, this study identifies the key EXP genes involved in strawberry fruit softening and reveals a regulatory role of histone modifications in their transcriptional regulation, which would facilitate functional studies of the EXP genes in the ripening of non-climacteric fruits.
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Fragaria , Fragaria/genética , Frutas , Regulação da Expressão Gênica de Plantas , Código das Histonas , FilogeniaRESUMO
BACKGROUND: Remote ischemic conditioning (RIC) by brief periods of limb ischemia and reperfusion protects against ischemia-reperfusion injury. We studied the cardioprotective role of extracellular vesicles (EV)s released into the circulation after RIC and EV accumulation in injured myocardium. METHODS: We used plasma from healthy human volunteers before and after RIC (pre-PLA and post-PLA) to evaluate the transferability of RIC. Pre- and post-RIC plasma samples were separated into an EV enriched fraction (pre-EV + and post-EV +) and an EV poor fraction (pre-EV- and post-EV-) by size exclusion chromatography. Small non-coding RNAs from pre-EV + and post-EV + were purified and profiled by NanoString Technology. Infarct size was compared in Sprague-Dawley rat hearts perfused with isolated plasma and fractions in a Langendorff model. In addition, fluorescently labeled EVs were used to assess homing in an in vivo rat model. (ClinicalTrials.gov, number: NCT03380663) RESULTS: Post-PLA reduced infarct size by 15% points compared with Pre-PLA (55 ± 4% (n = 7) vs 70 ± 6% (n = 8), p = 0.03). Post-EV + reduced infarct size by 16% points compared with pre-EV + (53 ± 15% (n = 13) vs 68 ± 12% (n = 14), p = 0.03). Post-EV- did not affect infarct size compared to pre-EV- (64 ± 3% (n = 15) and 68 ± 10% (n = 16), p > 0.99). Three miRNAs (miR-16-5p, miR-144-3p and miR-451a) that target the mTOR pathway were significantly up-regulated in the post-EV + group. Labelled EVs accumulated more intensely in the infarct area than in sham hearts. CONCLUSION: Cardioprotection by RIC can be mediated by circulating EVs that accumulate in injured myocardium. The underlying mechanism involves modulation of EV miRNA that may promote cell survival during reperfusion.
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Braço/irrigação sanguínea , Vesículas Extracelulares/transplante , Precondicionamento Isquêmico , MicroRNAs/metabolismo , Infarto do Miocárdio/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Miocárdio/metabolismo , Animais , Modelos Animais de Doenças , Vesículas Extracelulares/genética , Vesículas Extracelulares/metabolismo , Regulação da Expressão Gênica , Voluntários Saudáveis , Humanos , Preparação de Coração Isolado , Masculino , MicroRNAs/genética , Infarto do Miocárdio/genética , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/patologia , Traumatismo por Reperfusão Miocárdica/genética , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Miocárdio/patologia , Ratos Sprague-Dawley , Fluxo Sanguíneo RegionalRESUMO
Signal plasticity can maximize the usefulness of costly animal signals such as the electric organ discharges (EODs) of weakly electric fishes. Some species of the order Gymnotiformes rapidly alter their EOD amplitude and duration in response to circadian cues and social stimuli. How this plasticity is maintained across related species with different degrees of signal complexity is poorly understood. In one genus of weakly electric gymnotiform fish (Brachyhypopomus), only one species, B. bennetti, produces a monophasic signal while all other species emit complex biphasic or multiphasic EOD waveforms produced by two overlapping but asynchronous action potentials in each electric organ cell (electrocyte). One consequence of this signal complexity is the suppression of low-frequency signal content that is detectable by electroreceptive predators. In complex EODs, reduction of the EOD amplitude and duration during daytime inactivity can decrease both predation risk and the metabolic cost of EOD generation. We compared EOD plasticity and its underlying physiology in Brachyhypopomus focusing on B. bennetti. We found that B. bennetti exhibits minimal EOD plasticity, but that its electrocytes retained vestigial mechanisms of biphasic signaling and vestigial mechanisms for modulating the EOD amplitude. These results suggest that this species represents a transitional phenotypic state within a clade where signal complexity and plasticity were initially gained and then lost. Signal mimicry, mate recognition and sexual selection are potential factors maintaining the monophasic EOD phenotype in the face of detection by electroreceptive predators.
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Peixe Elétrico , Gimnotiformes , Potenciais de Ação , Animais , Órgão Elétrico , Transdução de SinaisRESUMO
Genome function is dynamically regulated in part by chromatin, which consists of the histones, non-histone proteins and RNA molecules that package DNA. Studies in Caenorhabditis elegans and Drosophila melanogaster have contributed substantially to our understanding of molecular mechanisms of genome function in humans, and have revealed conservation of chromatin components and mechanisms. Nevertheless, the three organisms have markedly different genome sizes, chromosome architecture and gene organization. On human and fly chromosomes, for example, pericentric heterochromatin flanks single centromeres, whereas worm chromosomes have dispersed heterochromatin-like regions enriched in the distal chromosomal 'arms', and centromeres distributed along their lengths. To systematically investigate chromatin organization and associated gene regulation across species, we generated and analysed a large collection of genome-wide chromatin data sets from cell lines and developmental stages in worm, fly and human. Here we present over 800 new data sets from our ENCODE and modENCODE consortia, bringing the total to over 1,400. Comparison of combinatorial patterns of histone modifications, nuclear lamina-associated domains, organization of large-scale topological domains, chromatin environment at promoters and enhancers, nucleosome positioning, and DNA replication patterns reveals many conserved features of chromatin organization among the three organisms. We also find notable differences in the composition and locations of repressive chromatin. These data sets and analyses provide a rich resource for comparative and species-specific investigations of chromatin composition, organization and function.
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Caenorhabditis elegans/citologia , Caenorhabditis elegans/genética , Cromatina/genética , Cromatina/metabolismo , Drosophila melanogaster/citologia , Drosophila melanogaster/genética , Animais , Linhagem Celular , Centrômero/genética , Centrômero/metabolismo , Cromatina/química , Montagem e Desmontagem da Cromatina/genética , Replicação do DNA/genética , Elementos Facilitadores Genéticos/genética , Epigênese Genética , Heterocromatina/química , Heterocromatina/genética , Heterocromatina/metabolismo , Histonas/química , Histonas/metabolismo , Humanos , Anotação de Sequência Molecular , Lâmina Nuclear/metabolismo , Nucleossomos/química , Nucleossomos/genética , Nucleossomos/metabolismo , Regiões Promotoras Genéticas/genética , Especificidade da EspécieRESUMO
Vesicles (Ves) within fungal cells are the critical linkage between intracellular and extracellular systems. This study explored the application of Pb2+ to probe the physiology of intracellular Ves in Rhodotorula mucilaginosa (Rho). At low Pb2+ levels (0-500 mg/L), there was no evident change in the content of extracellular polymeric substances (EPS) or microbial activity. At medium-high levels (1000-2000 mg/L), the sizes of Ves within the Rho cells were significantly enlarged, with abundant lead nano-particles (Pb NPs) formed either on the cell surface or interior, whereas the EPS content and bioactivity were still stable. At a high level (2500 mg/L), the Rho cells were severely deformed, with cell counts reduced by more than 99%. However, the EPS contents and the respiration rate of the surviving cells dramatically increased to the maximum values (i.e., 1785 mg/1010 cells and 37 mg C 10-10 cells h-1, respectively). The Ves surface adsorbed Pb cations with higher density, compared with the cell membrane. Moreover, fusion of some Ves to the membrane (functioning in transport) was observed under transmission electron microscope (TEM). Three pathways of detoxification via intracellular Ves were finally proposed, i.e., Ve-mediated transport (from intracellular to extracellular) of EPS components, absorption of Pb NPs on the Ve surface, and accumulation of Pb NPs within Ves. This study sheds light on the possibility of exploring microbial physiology via Pb2+ cations.
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Substâncias Perigosas/toxicidade , Chumbo/toxicidade , Rhodotorula/fisiologia , Adsorção , Cátions , Testes de ToxicidadeRESUMO
Long noncoding RNAs (lncRNAs) have been proved to play important roles in carcinogenesis and development of numerous cancers, but their biological functions in bladder cancer remain largely unknown. In this study, a novel lncRNA termed GAS6-AS2 were primary identified, and its roles as well as mechanisms in regulating proliferation and metastasis of bladder cancer cells were investigated. Clinically, GAS6-AS2 was significantly up-regulated in bladder cancer tissues and positively correlated with tumour stages and poor prognosis. Moreover, expression of GAS6-AS2 was also increased in bladder cancer cells compared with normal bladder cells. Further investigating the roles of GAS6-AS2, we found GAS6-AS2 regulated proliferation and proliferative activity of bladder cancer cells via inducing G1 phase arrest. What's more, we found that GAS6-AS2 contributed to metastatic abilities of cells. In mechanism, GAS6-AS2 could function as a competitive endogenous RNA (ceRNA) via direct sponging miR-298, which further regulating the expression of CDK9. Finally, we also proved that GAS6-AS2 knockdown suppressed tumour growth and metastasis in vivo. In conclusion, our study proved that GAS6-AS2 could function as a ceRNA and promote the proliferation and metastasis of bladder cancer cells, which provided a novel prognostic marker for bladder cancer patients in clinic.
Assuntos
Quinase 9 Dependente de Ciclina/genética , Regulação Neoplásica da Expressão Gênica , Peptídeos e Proteínas de Sinalização Intercelular/genética , Neoplasias Pulmonares/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , Neoplasias da Bexiga Urinária/genética , Animais , Pareamento de Bases , Sequência de Bases , Estudos de Casos e Controles , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Sobrevivência Celular , Quinase 9 Dependente de Ciclina/metabolismo , Feminino , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/secundário , Masculino , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/metabolismo , Pessoa de Meia-Idade , RNA Longo não Codificante/antagonistas & inibidores , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Análise de Sobrevida , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/mortalidade , Neoplasias da Bexiga Urinária/patologia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Expanded porphyrins have been attracting increasing attention owing to their unique optical and electrochemical properties as well as switchable aromaticity. Toward material applications, regioselective functionalization of the expanded porphyrins at their periphery is indeed challenging due to the presence of multiple reactive sites. Herein, a set of regioselective halogenated isomers (L5-Br-A/B/C) of neo-confused isosmaragdyrin (L5) are synthesized by a combination of the halogenation reaction of L5 and sequential macrocycle-to-macrocycle transformation reactions of its halogenated isomers. On this basis, the regioselectively functionalized isosmaragdyrins are utilized as building blocks for constructing multichromophoric porphyrinoids, specifically, heterodyads L5-ZnP-A/B/C, in which a common zinc porphyrin is linked at three different pyrrolic positions of isosmaragdyrins, respectively, by Sonogashira coupling reactions. The highly efficient energy cascade from porphyrin to isosmaragdyrin is elucidated using steady-state/time-resolved spectroscopies and theoretical calculations. Notably, the energy transfer processes from the porphyrin to the isosmaragdyrin moieties as well as the excitation energy transfer rates in L5-ZnP-A/B/C are highly dependent on the linking sites by through-bond and Förster-type resonance energy transfer mechanisms. The site-selective functionalization and subsequent construction of a set of heterodyads of the expanded porphyrinoid would provide opportunities for developing new materials for optoelectronic applications.
RESUMO
Although exogenous applications of gibberellins (GAs) delay tomato ripening, the regulatory mechanisms of GAs in the process have never been well recognized. Here, we report that the concentration of endogenous GAs is declined before the increase of ethylene production in mature-green to breaker stage fruits. We further demonstrate that reductions in GA levels via overexpression of a GA catabolism gene SlGA2ox1 specifically in fruit tissues lead to early ripening. Consistently, we have also observed that application of a GA biosynthetic inhibitor, prohexadione-calcium, at the mature-green stage accelerates fruit ripening, while exogenous GA3 application delays the process. Furthermore, we demonstrate that ethylene biosynthetic gene expressions and ethylene production are activated prematurely in GA-deficient fruits but delayed/reduced in exogenous GA3-treated WT fruits. We also show that the GA deficiency-mediated activation of ethylene biosynthesis is due to the activation of the ripening regulator genes RIN, NOR and CNR. In conclusion, our results demonstrate that GAs play a negative role in tomato fruit ripening.