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BACKGROUND: The PD-L1 on tumor cell-derived small extracellular vesicles (sEVs) can suppress the proliferation and cytokine production of T cells. However, PD-L1 can also be expressed by non-tumor cells. The present study is designed to test whether immunocytes release immunosuppressive PD-L1-positive sEVs. METHODS: sEVs were isolated from different clinical samples of head and neck squamous cell carcinoma (HNSCC) patients, the level and cellular origins of PD-L1-positive sEVs were assessed. Co-expression of CD80 on PD-L1-positive sEVs was examined to evaluate the immunosuppressive and tumor-promotive effects. RESULTS: PD-L1-positive sEVs in HNSCC patients had various cellular origins, including tumor cell, T cell, B cell, dendritic cell and monocyte/macrophage. However, PD-L1-positive sEVs derived from immune cells did not exert immunosuppressive functions due to the co-expression of CD80. It was verified that co-expression of CD80 disrupted the binding of sEV PD-L1 to its receptor PD-1 on T cells and attenuated the immunosuppression mediated by sEV PD-L1 both in vitro and in vivo. CONCLUSION: The study suggests that PD-L1-positive sEVs have the cellular origin and functional heterogeneity. Co-expression of CD80 could restrict the immunosuppressive effect of sEV PD-L1. A greater understanding of PD-L1-positive sEV subsets is required to further improve their clinical application.
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Vesículas Extracelulares , Neoplasias de Cabeça e Pescoço , Humanos , Carcinoma de Células Escamosas de Cabeça e Pescoço , Antígeno B7-H1/metabolismo , Linfócitos T , Vesículas Extracelulares/metabolismoRESUMO
BACKGROUND: An accurate intraoperative prediction of lymph node metastatic risk can help surgeons in choosing precise surgical procedures. We aimed to develop and validate nomograms to intraoperatively predict patterns of regional lymph node (LN) metastasis in patients with esophageal cancer. METHODS: The prediction model was developed in a training cohort consisting of 487 patients diagnosed with esophageal cancer who underwent esophagectomy with complete LN dissection from January 2016 to December 2016. Univariate and multivariable logistic regression were used to identify independent risk factors that were incorporated into a prediction model and used to construct a nomogram. Contrast-enhanced computed tomography reported LN status and was an important comparative factor of clinical usefulness in a validation cohort. Nomogram performance was assessed in terms of calibration, discrimination, and clinical usefulness. An independent validation cohort comprised 206 consecutive patients from January 2017 to December 2017. RESULTS: Univariate analysis and multivariable logistic regression revealed three independent predictors of metastatic regional LNs, three independent predictors of continuous regional LNs, and two independent predictors of skipping regional LNs. Independent predictors were used to build three individualized prediction nomograms. The models showed good calibration and discrimination, with area under the curve (AUC) values of 0.737, 0.738, and 0.707. Application of the nomogram in the validation cohort yielded good calibration and discrimination, with AUC values of 0.728, 0.668, and 0.657. Decision curve analysis demonstrated that the three nomograms were clinically useful in the validation cohort. CONCLUSION: This study presents three nomograms that incorporate clinicopathologic factors, which can be used to facilitate the intraoperative prediction of metastatic regional LN patterns in patients with esophageal cancer.
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Adenocarcinoma/secundário , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/secundário , Linfonodos/patologia , Nomogramas , Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Área Sob a Curva , Neoplasias Esofágicas/diagnóstico por imagem , Neoplasias Esofágicas/cirurgia , Carcinoma de Células Escamosas do Esôfago/diagnóstico por imagem , Carcinoma de Células Escamosas do Esôfago/cirurgia , Feminino , Seguimentos , Humanos , Linfonodos/diagnóstico por imagem , Linfonodos/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Retrospectivos , Tomografia Computadorizada por Raios XRESUMO
Influenza is an acute viral respiratory disease that can also cause gastroenteritis-like symptoms, such as abdominal pain, nausea, vomiting, and diarrhea. Immune dysfunction of adipose tissue is involved in the occurrence and prognosis of influenza viral pneumonia. In this study, we analyzed intestinal and adipose immune responses in mice infected with influenza virus and found that the impairment of intestinal and adipose immunity to influenza virus infection could be reversed by treatment with puerarin, a medicinal compound isolated from Pueraria lobata (called "gegen" in Chinese). We found that the lungs, small intestines (duodenum, ileum, jejunum) and large intestines (colon and rectum) of infected mice showed obvious inflammatory lesions, with significantly increased levels of virus, inflammatory cytokines (interleukin [IL]-6, IL-17, and tumor necrosis factor-α), Toll-like receptors 3, 4, and 9, and integrin αvß3 and α4, and a decreased level of secreted IgA compared to the normal control group (NC) (P < 0.05-0.001). Influenza virus infected mesenteric lymph nodes and adipose tissue, and adipokines (leptin, visfatin, "chemerin", and adiponectin) of lung and mesenteric adipose tissue were dysregulated. Puerarin treatment reversed the impairment of the intestinal and adipose immune responses in mice infected with influenza virus. Our findings suggest that influenza virus can infect adipose tissue and lead to intestinal adipose immune dysfunction in normal-weight mice and that the impairment of the intestinal and adipose immune response to influenza virus infection can be reversed by puerarin treatment.
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Tecido Adiposo/imunologia , Imunidade/efeitos dos fármacos , Intestinos/imunologia , Infecções por Orthomyxoviridae/tratamento farmacológico , Infecções por Orthomyxoviridae/imunologia , Vasodilatadores/farmacologia , Animais , Citocinas/metabolismo , Feminino , Humanos , Imunoglobulina A Secretora , Vírus da Influenza A Subtipo H1N1 , Influenza Humana/imunologia , Intestinos/patologia , Intestinos/virologia , Isoflavonas , Pulmão/patologia , Pulmão/virologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Nicotinamida Fosforribosiltransferase , Pneumonia , Pueraria/química , RNA Viral , Carga ViralRESUMO
Puerarin is a major isofiavone compound isolated from the root of Pueraria lobata. It was reported that puerarin had antioxidant, antiinflammatory, antitumor, cholesterol lowering, liver protective, and neuroprotective properties. However, few studies have explored the antiviral effect of puerarin and its target mechanism related to influenza virus. Here, the antiinfluenza activity of puerarin in vitro and in vivo and its mode of action on the potential inhibition of neuraminidase (NA) were investigated. Puerarin displayed an inhibitory effect on A/FM/1/1947(H1N1) (EC50 = 52.06 µM). An indirect immunofluorescence assay indicated that puerarin blocked the nuclear export of viral NP. The inhibition of NA activity confirmed that puerarin can block the release of newly formed virus particles from infected cells. Puerarin (100 and 200 mg/kg/d) exhibited effective antiviral activity in mice, conferring 50% and 70% protection from death against H1N1, reducing virus titers, and effectively alleviating inflammation in the lungs. The molecular docking results showed that puerarin had a strong binding affinity with NA from H1N1. The results of the molecular dynamics simulation revealed that puerarin had higher stable binding at the 150-loop region of the NA protein. These results demonstrated that puerarin acts as a NA blocker to inhibit influenza A virus both in cellular and animal models. Thus, puerarin has potential utility for the treatment of the influenza virus infection.
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Antivirais/farmacologia , Isoflavonas/farmacologia , Neuraminidase/antagonistas & inibidores , Infecções por Orthomyxoviridae/tratamento farmacológico , Proteínas Virais/antagonistas & inibidores , Animais , Cães , Feminino , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Células Madin Darby de Rim Canino , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Simulação de Acoplamento Molecular , Carga Viral/efeitos dos fármacosRESUMO
With the increased availability of highly maneuverable unmanned surface/underwater vehicles, abundant ocean data can now be collected. This study uses tomographic techniques to extend the survey area covered by moving vehicles. An acoustic reciprocal transmission experiment was conducted using three tomographic sensors installed on an autonomous underwater vehicle, a fishing ship, and a buoy. The distributed sensing method is applied for currents estimation. The estimated currents near the ship show consistent results with the direct measurements. In particular, an anticyclonic circulation was revealed. Further, a general least-squares method is employed to improve the estimate of this vortical structure.
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AIMS: The purpose of this study was to explore the potential involvement of Fra-1, c-Jun and c-Fos, three vital members of the AP-1 complex, in the pathogenesis of odontogenic keratocysts (OKCs). METHODS AND RESULTS: Tissue samples, containing 10 normal oral mucosa (OM), 10 dentigerous cysts (DC) and 32 OKC specimens, were applied to investigate the expression levels of Fra-1, c-Jun and c-Fos by immunohistochemistry and real-time-quantitative polymerase chain reaction (RT-qPCR). The association between Fra-1, c-Jun and c-Fos expression levels and markers of proliferation [Ki-67, proliferating cell nuclear antigen (PCNA)], anti-apoptosis (Bcl-2) was then investigated in the OKC serial tissue sections. The results showed that Fra-1, c-Jun and c-Fos expression levels were increased significantly in OKCs compared to these in OM and DC tissue samples. Meanwhile, the expression levels of Fra-1, c-Jun and c-Fos were associated positively with the expression levels of Ki-67, PCNA and Bcl-2, as confirmed further by double-labelling immunofluorescence analysis and hierarchical analysis. CONCLUSIONS: This study revealed for the first time that Fra-1, c-Jun and c-Fos were overexpressed in OKCs and had a close correlation with proliferation and anti-apoptosis potential of OKCs.
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Apoptose/fisiologia , Proliferação de Células/fisiologia , Mucosa Bucal/metabolismo , Cistos Odontogênicos/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Proteínas Proto-Oncogênicas c-jun/metabolismo , Humanos , Imuno-Histoquímica , Mucosa Bucal/patologia , Cistos Odontogênicos/patologiaRESUMO
Phospholipase D (PLD) hydrolyzes phospholipids to generate a free polar head group (e.g., choline) and a second messenger phosphatidic acid and plays diverse roles in plant growth and development, including seed germination, leaf senescence, root hair growth, and hypocotyl elongation. However, the function of PLD in cotton remains largely unexplored. Here, the comprehensive molecular characterization of GhPLDα1 was explored with its role in upland cotton (Gossypium hirsutum) fiber development. The GhPLDα1 gene was cloned successfully, and a sequence alignment showed that GhPLDα1 contains one C2 domain and two HKD (HxKxxxxD) domains. Quantitative reverse transcriptase-polymerase chain reaction measured the expression of GhPLDα1 in various cotton tissues with the highest level in fibers at 20 days post anthesis (d.p.a.). Fluorescent microscopy and immunoblotting in tobacco epidermis showed the GhPLDα1 distribution in both cell membranes and the cytoplasm. An activity assay indicated changes in PLDα enzyme activity in developing fiber cells with a peak level at 20 d.p.a., coinciding with the onset of cellulose accumulation and the increased H2O2 content during fiber development. Furthermore, the inhibition of PLDα activity obviously decreased the cellulose and H2O2 contents of in vitro-cultured cotton fibers. These results provide important evidence explaining the relationship of GhPLDα1 with secondary cell wall thickening in cotton fibers in that GhPLDα1 may correlate with the increased H2O2 content at the onset of secondary cell wall thickening, ultimately promoting cellulose biosynthesis.
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Parede Celular , Fibra de Algodão , Fosfolipase D/genética , Regulação da Expressão Gênica de Plantas , Humanos , Microscopia de FluorescênciaRESUMO
Intergenic transcription by RNA Polymerase II (Pol II) is widespread in plant and animal genomes, but the functions of intergenic transcription or the resulting noncoding transcripts are poorly understood. Here, we show that Arabidopsis Pol II is indispensable for endogenous siRNA-mediated transcriptional gene silencing (TGS) at intergenic low-copy-number loci, despite the presence of two other polymerases-Pol IV and Pol V-that specialize in TGS through siRNAs. We show that Pol II produces noncoding scaffold transcripts that originate outside of heterochromatic, siRNA-generating loci. Through these transcripts and physical interactions with the siRNA effector protein ARGONAUTE4 (AGO4), Pol II recruits AGO4/siRNAs to homologous loci to result in TGS. Meanwhile, Pol II transcription also recruits Pol IV and Pol V to different locations at heterochromatic loci to promote siRNA biogenesis and siRNA-mediated TGS, respectively. This study establishes that intergenic transcription by Pol II is required for siRNA-mediated TGS, and reveals an intricate collaboration and division of labor among the three polymerases in gene silencing.
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Arabidopsis/enzimologia , Arabidopsis/genética , DNA Intergênico/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Inativação Gênica , RNA Polimerase II/metabolismo , RNA Interferente Pequeno/metabolismo , Sequência de Aminoácidos , Cromatina/metabolismo , RNA Polimerases Dirigidas por DNA/química , RNA Polimerases Dirigidas por DNA/genética , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Mutação , Alinhamento de SequênciaRESUMO
MAIN CONCLUSION: Cotton cytosolic pyruvate kinase GhPK6 is preferentially expressed in the late stage of fiber elongation process, transgenic experiments indicated that its expression level was negatively correlated to cell expansion rate. Pyruvate kinase (PK) plays vital regulatory roles in rapid cell growth in mammals. However, the function of PK in plant cell growth remains unclear. In allotetraploid upland cotton (Gossypium hirsutum L.), a total of 33 PK genes are encoded by the genome. Analysis of the transcriptome data indicated that only two cytosolic PK genes, GhPK6 and its duplicated gene GhPK26, are preferentially expressed in elongating cotton fiber cells. RT-qPCR and western blot analyses revealed that the expression of GhPK6 was negatively correlated with fiber elongation rate, which well explains the observed sharp increase of cytosolic PK activity at the end of fast fiber elongation process. Furthermore, virus-induced gene silencing of GhPK6 in cotton plants resulted in increased fiber cell elongation and reduced reactive oxygen species (ROS) accumulation. On the contrary, Arabidopsis plants ectopically expressing GhPK6 exhibited ROS-mediated growth inhibition, whereas the addition of ROS scavenging reagents could partly rescue this inhibition. These data collectively suggested that GhPK6 might play an important role in regulating cotton fiber elongation in a ROS-dependent inhibition manner.
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Fibra de Algodão , Citosol/enzimologia , Gossypium/enzimologia , Proteínas de Plantas/metabolismo , Piruvato Quinase/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Sequência de Bases , Western Blotting , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genoma de Planta/genética , Gossypium/genética , Gossypium/crescimento & desenvolvimento , Família Multigênica/genética , Proteínas de Plantas/genética , Piruvato Quinase/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência do Ácido Nucleico , TetraploidiaRESUMO
BACKGROUND: Plenty of proteomic studies were performed to characterize the allotetraploid upland cotton fiber elongation process, whereas little is known about the elongating diploid cotton fiber proteome. METHODS: In this study, we used a two-dimensional electrophoresis-based comparative proteomic approach to profile dynamic proteomes of diploid Asian cotton ovules with attached fibers in the early stages of fiber elongation process. One-way ANOVA and Student-Newman-Keuls test were used to find the differentially displayed protein (DDP) spots. RESULTS: A total of 55 protein spots were found having different abundance ranging from 1 to 9 days post-anthesis (DPA) in a two-day interval. These 55 DDP spots were all successfully identified using high-resolution mass spectrometric analyses. Gene ontology analyses revealed that proteoforms involved in energy/carbohydrate metabolism, redox homeostasis, and protein metabolism are the most abundant. In addition, orthologues of the 13 DDP spots were also found in differential proteome of allotetraploid elongating cotton fibers, suggesting their possible essential roles in fiber elongation process. CONCLUSIONS: Our results not only revealed the dynamic proteome change of diploid Asian cotton fiber and ovule during early stages of fiber elongation process but also provided valuable resource for future studies on the molecular mechanism how the polyploidization improves the trait of fiber length.
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Obtaining the horizontal variation of temperature and current fields of a water column usually requires travel-time measurements of acoustic signals traveling along different paths between several horizontally distributed transceivers. This study explores the possibility of using a pair of transceivers deployed in a highly-reverberant harbor environment to extract spatial information of the water. Multipath acoustic propagation of two main arrival groups, i.e., direct arrivals and arrivals reflecting off the harbor side, was observed in the pulse responses measured in the harbor environment during the flood tide. Compared with the direct point measurements of temperature and current, the path-averaged measurements show a similar temporal variation during the experiment, demonstrating the possibility of estimating the spatial variation of the currents and temperatures using the multipath acoustic propagation.
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Recently, the diagnosis of the characteristic of pulsed underwater electrical discharges plasma have received significant attention. The measurement of a time-spatial resolved spectrum emitted from a single discharge pulse is important for understanding the time-spatial evolution characteristics of plasma generated by a pulsed high-voltage discharge in water. In this paper, a high speed time-spatial resolvable spectrograph for measuring the emission spectrum of a single electrical discharge pulse was reported. The high speed time-spatial resolvable spectrograph has been constructed by combining an ultrahigh-speed frame camera system with monochromator. Software for the spectral analyzing was also developed. The performance of the spectrograph was tested by using a 632.8 nm He-Ne laser beam at a 1 200 g x mm(-1) grating. The pixel resolution for 632.8 nm spectra is 0.013 nm. The instrument broadening for 632.8 nm spectra is (0.150 ± 0.009)nm when the exposure.time of the camera is 20 ns and the width of entrance slit is 0.2 mm, and increases with increasing the slit width. The change of exposure time of the camera has no influence on the instrument broadening, ensuring the spectrograph in a steady performance while adjusting the exposure time of the camera. With the spectrograph, time-spatial resolved spectra emitted from a single discharge pulse of an underwater nanoseconds spark discharge were obtained. It provides good data for investigating the time-spatial evolution characteristics of the discharge plasma during a single discharge pulse. The spectrograph developed in this work provides a technical approach for studying the time-spatial evolution characteristic of, plasma generated by a single electrical discharge pulse.
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OBJECTIVE: To analyze and compare the clinical effects of femoral neck dynamic cross screw system (FNS) and cannulated screws(CS) in the treatment of vertically unstable femoral neck fractures. METHODS: The clinical data and short-term follow-up results of 40 patients with vertically unstable femoral neck fractures admitted from July 2020 to August 2021 were retrospectively analyzed. According to different internal fixation methods, 40 patients were divided into two groups, 20 cases in FNS group included 11 males and 9 females with a median of 58.5(50.3, 62.5) years old, and 20 in CS group included 9 males and 11 females with a median of 52.0(40.5, 58.0) years old. The operation time, knife edge length, blood loss and treatment cost of two gruops were observed and compared. The postoperative fracture healing and internal fixation were evaluated with X-ray imaging data, and the femoral neck shortening of the affected side was measured. The incidence of thigh irritation, the time of partial weight bearing and full weight bearing, early necrosis of femoral head, reoperation revision and Harris scores were compared between two groups. RESULTS: FNS group was followed up for 18.0(15.0, 19.0) months, CS group for 17.0(15.0, 18.8) months. There was no significant difference in operation time, incision length and blood loss between two groups(P>0.05). The cost of diagnosis and treatment in FNS group was higher than that in CS group(P<0.001). In FNS group, there was no irritation sign of the affected side thigh, while in CS group, there were 6 cases with discomfort or irritation sign of the lateral thigh(P<0.05). The average time of partial weight bearing activity in CS group was later than that in FNS group(P<0.05); However, there was no significant difference in the activity time of complete weight bearing between two groups(P=0.011>0.05). At the last follow-up, the shortened length of the affected femoral neck in CS group was greater than that in FNS group(P<0.05). There was no early necrosis of femoral head and reoperation in both groups. There was no significant difference in Harris score between two groups 12 months after operation(P>0.05). CONCLUSION: FNS treatment of vertically unstable femoral neck fractures can significantly reduce the incidence of lateral thigh irritation sign, and effectively reduce the postoperative shortening rate of vertically unstable femoral neck fractures, which can provide a relatively stable anti rotation force and anti cutting force, so that patients can go to the ground relatively early, which is conducive to the recovery of the affected hip joint function after surgery. It is a new option for the surgical treatment of vertically unstable femoral neck fractures. However, due to the high cost of treatment, In clinical practice, appropriate surgical treatment is selected according to the actual situation.
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Parafusos Ósseos , Fraturas do Colo Femoral , Fixação Interna de Fraturas , Humanos , Fraturas do Colo Femoral/cirurgia , Masculino , Feminino , Pessoa de Meia-Idade , Fixação Interna de Fraturas/métodos , Fixação Interna de Fraturas/instrumentação , Seguimentos , Estudos Retrospectivos , AdultoRESUMO
BACKGROUND: Mitochondrial dysfunction plays a vital role in the progression of vascular dementia (VaD). We hypothesized that transfer of exogenous mitochondria might be a beneficial strategy for VaD treatment. OBJECTIVE: The study was aimed to investigate the role of mitochondrial therapy in cognitive function of VaD. METHODS: The activity and integrity of isolated mitochondria were detected using MitoTracker and Janus Green B staining assays. After VaD mice were intravenously injected with exogenous mitochondria, Morris water maze and passive avoidance tests were used to detect cognitive function of VaD mice. Haematoxylin and eosin, Nissl, TUNEL, and Golgi staining assays were utilized to measure neuronal and synaptic injury in the hippocampus of VaD mice. Detection kits were performed to detect mitochondrial membrane potential (ΔΨ), SOD activity and the levels of ATP, ROS, and MDA in the brains of VaD mice. RESULTS: The results showed that isolated mitochondria were intact and active. Mitochondrial therapy could ameliorate cognitive performance of VaD mice. Additionally, mitochondrial administration could attenuate hippocampal neuronal and synaptic injury, improve mitochondrial ΔΨ, ATP level and SOD activity, and reduce ROS and MDA levels in the brains of VaD mice. CONCLUSIONS: The study reports profitable effect of mitochondrial therapy against cognitive impairment of VaD, making mitochondrial treatment become a promising therapeutic strategy for VaD.
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Disfunção Cognitiva , Demência Vascular , Camundongos , Animais , Demência Vascular/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Cognição , Disfunção Cognitiva/metabolismo , Superóxido Dismutase/metabolismo , Mitocôndrias , Trifosfato de Adenosina/metabolismo , Aprendizagem em Labirinto/fisiologia , Hipocampo/metabolismoRESUMO
In our previous study, we used a comparative proteomic approach based on 2DE to profile dynamic proteomes of cotton fibers and found 235 protein spots differentially expressed during the elongation process ranging from 5 to 25 days post-anthesis. Of them, only 106 differentially expressed proteins (DEPs) were identified by MS due to database limitations at the time. In the present work, we successfully identified the remaining 129 DEPs from the same experimental system using high-resolution MS with an updated database. Bioinformatic analysis revealed that proteins involved in carbohydrate and protein metabolism, transport, and redox homeostasis are the most abundant, and glycolysis was found to be the most significantly regulated process during fiber elongation. Our high-confidence reference dataset, composed of 235 DEPs, provides a valuable resource for future studies on the molecular mechanism of cotton fiber elongation.
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Fibra de Algodão , Bases de Dados de Proteínas , Perfilação da Expressão Gênica , Gossypium/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteômica , Eletroforese em Gel Bidimensional , Regulação da Expressão Gênica de Plantas , Gossypium/citologia , Gossypium/genética , Redes e Vias Metabólicas , Padrões de ReferênciaRESUMO
BACKGROUND: MicroRNAs (miRNAs) and other types of small regulatory RNAs play critical roles in the regulation of gene expression at the post-transcriptional level in plants. Cotton is one of the most economically important crops, but little is known about the roles of miRNAs during cotton fiber elongation. RESULTS: Here, we combined high-throughput sequencing with computational analysis to identify small RNAs (sRNAs) related to cotton fiber elongation in Gossypium hirsutum L. (G. hirsutum). The sequence analysis confirmed the expression of 79 known miRNA families in elongating fiber cells and identified 257 novel miRNAs, primarily derived from corresponding specific loci in the Gossypium raimondii Ulbr. (G. raimondii) genome. Furthermore, a comparison of the miRNAomes revealed that 46 miRNA families were differentially expressed throughout the elongation period. Importantly, the predicted and experimentally validated targets of eight miRNAs were associated with fiber elongation, with obvious functional relationships with calcium and auxin signal transduction, fatty acid metabolism, anthocyanin synthesis and the xylem tissue differentiation. Moreover, one tasiRNA was also identified, and its target, ARF4, was experimentally validated in vivo. CONCLUSION: This study not only facilitated the discovery of 257 novel low-abundance miRNAs in elongating cotton fiber cells but also revealed a potential regulatory network of nine sRNAs important for fiber elongation. The identification and characterization of miRNAs in elongating cotton fiber cells might promote the further study of fiber miRNA regulation mechanisms and provide insight into the importance of miRNAs in cotton.
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Fibra de Algodão , Gossypium/genética , MicroRNAs/genética , RNA de Plantas/genética , Sequência de Bases , Biblioteca Gênica , Redes Reguladoras de Genes , Gossypium/crescimento & desenvolvimento , Sequenciamento de Nucleotídeos em Larga Escala , Família Multigênica , Análise de Sequência de RNARESUMO
Plant microRNAs (miRNAs) have been shown to play critical roles in regulating gene expression at the post-transcriptional level. In this study, we employed high throughput sequencing combined with computational analysis to survey miRNAomes from the seedlings of rice under normal conditions and treatments of H(2)O(2) that result in oxidative stress. Comparison of the miRNAomes and subsequent northern blot analysis identified seven miRNA families differentially expressed under H(2)O(2) stress. Predicted and experimentally validated targets of these H(2)O(2)-responsive miRNAs are involved in different cellular responses and metabolic processes including transcriptional regulation, nutrient transport, auxin homeostasis, cell proliferation and programmed cell death. This indicates that diverse miRNAs form a complex regulatory network to coordinate plants' responses under oxidative stress. In addition, we also discovered 32 new miRNAs in the seedlings of rice. Interestingly, of these new miRNAs, miR3981 was originally found to be a putative exonic miRNA located in the exon of AK106348, suggesting that plants may also use some exons as an miRNA source. This study is the first genome-wide investigation of H(2)O(2)-regulated miRNAs in plants and broadens our perspectives on the important regulatory roles of miRNAs in plant oxidative stress and physiological adaption.
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MicroRNAs/metabolismo , Oryza/genética , Estresse Oxidativo , RNA de Plantas/metabolismo , Éxons , Sequenciamento de Nucleotídeos em Larga Escala , Peróxido de Hidrogênio/farmacologia , MicroRNAs/química , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , RNA de Plantas/química , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismoRESUMO
Distributed underwater sensors are expected to provide oceanographic monitoring over large areas. As fabrication technology advances, low cost sensors will be available for many uses. The sensors communicate to each other and are networked using acoustic communications. This paper first studies the performance of such systems for current measurements using tomographic inversion approaches to compare with that of a conventional system which distributes the sensors on the periphery of the area of interest. It then proposes two simple signal processing methods for ocean current mapping (using distributed networked sensors) aimed at real-time in-buoy processing. Tomographic inversion generally requires solving a challenging high dimensional inverse problem, involving substantial computations. Given distributed sensors, currents can be constructed locally based on data from neighboring sensors. It is shown using simulated data that similar results are obtained using distributed processing as using conventional tomographic approaches. The advantage for distributed systems is that by increasing the number of nodes, one gains a much more improved performance. Furthermore, distributed systems use much less energy than a conventional tomographic system for the same area coverage. Experimental data from an acoustic communication and networking experiment are used to demonstrate the feasibility of acoustic current mapping.
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Acústica/instrumentação , Monitoramento Ambiental/instrumentação , Água do Mar , Som , Transdutores , Algoritmos , Desenho de Equipamento , Estudos de Viabilidade , Movimento (Física) , Oceanos e Mares , Processamento de Sinais Assistido por Computador , Razão Sinal-Ruído , Espectrografia do Som , Fatores de TempoRESUMO
Enhancement of oxidative stress and resultant neuronal injury play important roles in initiating cognitive impairment during the aging process. Thus, attenuating oxidative injury is regarded as a profitable therapeutic strategy for age-associated cognitive impairment. Previous studies showed that gliclazide (Gli) had a protective role in neuronal injury from cerebral ischemia/reperfusion (I/R) injury. However, whether Gli has a profitable effect on age-associated cognitive impairment remains largely unclear. The present study showed that Gli held the potential to attenuate neuronal apoptosis in D-gal-induced senescent cells and aging mice. Additionally, Gli could alleviate synaptic injury and cognitive function in D-gal-induced aging mice. Further study showed that Gli could attenuate oxidative stress in D-gal-induced senescent cells and aging mice. The p38 MAPK pathway was predicted as the downstream target of Gli retarding oxidative stress using in silico analysis. Further studies revealed that Gli attenuated D-gal-induced phosphorylation of p38 and facilitated Nrf2 nuclear expression, indicating that the anti-oxidative property of Gli may be associated with the p38 MAPK pathway. The study demonstrates that Gli has a beneficial effect on ameliorating D-gal-induced neuronal injury and cognitive impairment, making this compound a promising agent for the prevention and treatment of age-associated cognitive impairment.
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The functional characterization of novel transcription factors identified by systematic analysis remains a major challenge due to insufficient data to interpret their specific roles in signaling networks. Here we present a DNA-binding sequence discovery method to in vitro identify a G-rich, 11-bp DNA-binding motif of a novel potential transcription factor AtYY1, a zinc finger protein in Arabidopsis, by using polymerase chain reaction-assisted in vitro selection and surface plasmon resonance analysis. Further mutational analysis of the conserved G bases of the potential motif confirmed that AtYY1 specifically bound to these conserved G sites. Additionally, genome-wide target gene analysis revealed that AtYY1 was involved in diverse cellular pathways, including glucose metabolism, photosynthesis, phototropism, and stress response.