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1.
Planta ; 253(2): 26, 2021 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-33410920

RESUMO

MAIN CONCLUSION: Circular RNAs (circRNAs) identification, expression profiles, and construction of circRNA-parental gene relationships and circRNA-miRNA-mRNA ceRNA networks indicate that circRNAs are involved in flag leaf senescence of rice. Circular RNAs (circRNAs) are a class of 3'-5' head-to-tail covalently closed non-coding RNAs which have been proved to play important roles in various biological processes. However, no systematic identification of circRNAs associated with leaf senescence in rice has been studied. In this study, a genome-wide high-throughput sequencing analysis was performed using rice flag leaves developing from normal to senescence. Here, a total of 6612 circRNAs were identified, among which, 113 circRNAs were differentially expressed (DE) during the leaf senescence process. Moreover, 4601 (69.59%) circRNAs were derived from the exons or introns of their parental genes, while 2110 (71%) of the parental genes produced only one circRNA. The sequence alignment analysis showed that hundreds of rice circRNAs were conserved among different plant species. Gene Ontology (GO) enrichment analysis revealed that parental genes of DE circRNAs were enriched in many biological processes closely related to leaf senescence. Through weighted gene co-expression network analysis (WGCNA), six continuously down-expressed circRNAs, 18 continuously up-expressed circRNAs and 15 turn-point high-expressed circRNAs were considered to be highly associated with leaf senescence. Additionally, a total of 17 senescence-associated circRNAs were predicted to have parental genes, in which, regulations of three circRNAs to their parental genes were validated by qRT-PCR. The competing endogenous RNA (ceRNA) networks were also constructed. And a total of 11 senescence-associated circRNAs were predicted to act as miRNA sponges to regulate mRNAs, in which, regulation of two circRNAs to eight mRNAs was validated by qRT-PCR. It is discussed that senescence-associated circRNAs were involved in flag leaf senescence probably through mediating their parental genes and ceRNA networks, to participate in several well-studied senescence-associated processes, mainly including the processes of transcription, translation, and posttranslational modification (especially protein glycosylation), oxidation-reduction process, involvement of senescence-associated genes, hormone signaling pathway, proteolysis, and DNA damage repair. This study not only showed the systematic identification of circRNAs involved in leaf senescence of rice, but also laid a foundation for functional research on candidate circRNAs.

2.
Biomed Res Int ; 2020: 8891876, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33381597

RESUMO

MicroRNA-361-5p (miR-361-5p) is a tumor suppressor miRNA that is dysregulated in several types of human cancer. However, the functional significance of miR-361-5p in hepatocellular carcinoma (HCC) is unclear. This study explored the biological function of miR-361-5p in regulating the progression of HCC and the underlying molecular mechanism. RT-qPCR analysis showed that miR-361-5p was downregulated in HCC tissues and cell lines. Functional analysis revealed that miR-361-5p acted as a tumor suppressor, inhibiting cell proliferation, migration, and invasion in HCC cell lines. Bioinformatics analyses identified Twist1 as a direct target of miR-361-5p, which was validated by dual-luciferase reporter assays, RT-qPCR, and western blotting. Rescue experiments indicated that Twist1 may mediate the tumor-suppressive effect of miR-361-5p in HCC cells, and this was supported by the effect of miR-361-5p on inhibiting the epithelial-mesenchymal transition (EMT) by targeting Twist1. This study is the first to suggest that miR-361-5p inhibits tumorigenesis and EMT in HCC by targeting Twist1. These findings are valuable for the diagnosis and clinical management of HCC.

3.
Mar Pollut Bull ; 160: 111168, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33181914

RESUMO

Climate Change solutions include CO2 extraction from atmosphere and water with burial by living habitats in sediment/soil. Nowhere on the planet are blue carbon plants which carry out massive carbon extraction and permanent burial more intensely concentrated than in SE Asia. For the first time we make a national and total inventory of data to date for "blue carbon" buried from mangroves and seagrass and delineate the constraints. For an area across Southeast Asia of approximately 12,000,000 km2, supporting mangrove forests (5,116,032 ha) and seagrass meadows (6,744,529 ha), we analyzed the region's current blue carbon stocks. This estimate was achieved by integrating the sum of estuarine in situ carbon stock measurements with the extent of mangroves and seagrass across each nation, then summed for the region. We found that mangroves ecosystems regionally supported the greater amount of organic carbon (3095.19Tg Corg in 1st meter) over that of seagrass (1683.97 Tg Corg in 1st meter), with corresponding stock densities ranging from 15 to 2205 Mg ha-1 and 31.3 to 2450 Mg ha-1 respectively, a likely underestimate for entire carbon including sediment depths. The largest carbon stocks are found within Indonesia, followed by the Philippines, Papua New Guinea, Myanmar, Malaysia, Thailand, Tropical China, Viet-Nam, and Cambodia. Compared to the blue carbon hotspot of tropical/subtropical Gulf of Mexico's total carbon stock (480.48 Tg Corg), Southeast Asia's greater mangrove-seagrass stock density appears a more intense Blue Carbon hotspot (4778.66 Tg Corg). All regional Southeast Asian nation states should assist in superior preservation and habitat restoration plus similar measures in the USA & Mexico for the Gulf of Mexico, as apparently these form two of the largest tropical carbon sinks within coastal waters. We hypothesize it is SE Asia's regionally unique oceanic-geologic conditions, placed squarely within the tropics, which are largely responsible for this blue carbon hotspot, that is, consistently high ambient light levels and year-long warm temperatures, together with consistently strong inflow of dissolved carbon dioxide and upwelling of nutrients across the shallow geological plates.


Assuntos
Sequestro de Carbono , Ecossistema , Ásia Sudeste , China , Golfo do México , Indonésia , Malásia , Papua Nova Guiné , Filipinas , Tailândia , Vietnã , Áreas Alagadas
4.
Environ Sci Technol ; 54(22): 14750-14760, 2020 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-33103882

RESUMO

Intensive macroalgal blooms, a source of labile organic carbon (LOC) induced by coastal nutrient loading in some seagrass ecosystems, create ideal conditions for enhanced recalcitrant organic carbon (ROC) loss via the cometabolism effect. Here, we carried out a 62-day laboratory experiment to see if density-dependent addition of macroalgal biomass can influence the seagrass decomposition process, including seagrass detritus carbon chemistry, greenhouse emissions, and bacterial communities. We found that higher density macroalgal addition stimulated microbes to decompose ∼20% more of the seagrass biomass compared to other treatments, which was also reflected in enhanced (∼twofold) greenhouse gas emissions. Although the composition of the seagrass-associated microbiome communities was unaffected by the addition of macroalgae, we showed that high macroalgal addition caused a relative depletion in the ROC as lignin and lipid compounds, as well as δ13C depletion and δ15N enrichment of the seagrass detritus. These results suggest that macroalgal blooms may stimulate the remineralization of recalcitrant components of seagrass detritus via cometabolism, possibly through providing available energy or resources for the synthesis of ROC-degrading enzymes within the resident microbial population. This study provides evidence that cometabolism can be a mechanism for leading to reduced seagrass blue carbon sequestration and preservation.

5.
Mar Pollut Bull ; 160: 111594, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32898739

RESUMO

Nitrogen mineralization is a critical biogeochemical process that transfers organic nitrogen into inorganic forms using heterotrophic microorganisms. However, few studies have focused on this potential nutrient supplier. In this study, the composition of sediment organic nitrogen (SON) was studied, and nitrogen mineralization flux entering the water column was quantified. The results indicate that acid-hydrolyzable nitrogen (AHN) accounts for more than 40% of the SON, especially in the riverine input and marine aquaculture areas, which had significantly higher concentrations than the bay mouth area. Similar results were found for the ammonium nitrogen (AN), amino-sugar nitrogen (ASN), the total hydrolyzable amino acid (THAA), and unidentified hydrolyzable nitrogen (HUN). The mineralization rate in the marine aquaculture area was as high as 9.03 ± 1.33 mg·kg-1·d-1, while those of the riverine input (4.77 ± 1.55 mg·kg-1·d-1) and bay mouth (5.12 ± 1.42 mg·kg-1·d-1) areas were lower. The SON fractions, including the AHN, AN, ASN, and AAN, could obviously affect the mineralization of the SON. However, the extracellular enzymes, including proteinase and urease, are the predominant factors controlling the SON mineralization process. Anthropogenic activities, including riverine input and marine aquaculture, exert significant influences on the fractions and mineralization of the SON, and thus, they may increase the amount of dissolved inorganic nitrogen in the bottom of the water column in Daya Bay.


Assuntos
Baías , Nitrogênio , Aquicultura , China , Monitoramento Ambiental , Sedimentos Geológicos , Nitrogênio/análise
7.
World J Surg Oncol ; 18(1): 235, 2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32883303

RESUMO

BACKGROUND: Several randomized controlled trials (RCTs) have compared the treatment of acute lung injury (ALI) with omega-3 fatty, yet the results remained inconsistent. Therefore, we attempted this meta-analysis to analyze the role of omega-3 fatty in the treatment of ALI patients. METHODS: We searched PubMed databases from inception date to October 31, 2019, for RCTs that compared the treatment of ALI with or without omega-3 fatty. Two authors independently screened the studies and extracted data from the published articles. Summary mean differences (MD) with 95% confidence intervals (CI) were calculated for each outcome by fixed- or random-effects model. RESULTS: Six RCTs with a total of 277 patients were identified, of whom 142 patients with omega-3 fatty acid treatment and 135 patients without omega-3 fatty treatment. Omega-3 fatty treatments significantly improve the PaO2 (MD = 13.82, 95% CI 8.55-19.09), PaO2/FiO2 (MD = 33.47, 95% CI 24.22-42.72), total protein (MD = 2.02, 95% CI 0.43-3.62) in ALI patients, and omega-3 fatty acid treatments reduced the duration of mechanical ventilation (MD = - 1.72, 95% CI - 2.84 to - 0.60) and intensive care unit stay (MD = - 1.29, 95% CI - 2.14 to - 0.43) in ALI patients. CONCLUSIONS: Omega-3 fatty can effectively improve the respiratory function and promote the recovery of ALI patients. Future studies focused on the long-term efficacy and safety of omega-3 fatty use for ALI are needed.

8.
Vet Microbiol ; 247: 108757, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32768210

RESUMO

The primary objective of this study was to determine the distribution of major pathogens causing mastitis and analyze the association of the main pathogens with the type of milk {milk samples collected from cows with mastitis of different severities, which consisted of subclinical mastitis (Lanzhou Mastitis Test weak positive, positive, and strong positive) and clinical mastitis}, region, season, bedding material, parity and lactation stage on large dairy farms. Fifteen large dairy farms in twelve major milk-producing provinces of China were enrolled in the study for approximately one year; 1,153 mastitis milk samples were collected and processed. The most frequently isolated pathogens were Staphylococcus spp. (39.03 %), Streptococcus spp. (11.01 %), Bacillus spp. (8.24 %), Aerococcus viridans (6.76 %), and Acinetobacter spp. (3.38 %), and most of these pathogens were environmental bacteria (67.53 %). Contagious pathogens (Staphylococcus aureus and Streptococcus agalactiae) were more prevalent in milk samples from bovine clinical mastitis cases than in milk samples from bovine subclinical mastitis cases. The percentages of Staphylococcus spp. and Bacillus spp. might be higher in northeastern farms than in farms located in other regions. A higher percentage of Staphylococcus spp. was observed in summer, while a lower proportion of Streptococcus spp. was detected in fall. Streptococcus agalactiae and Streptococcus uberis were more frequently isolated in farms using sand bedding, whereas Staphylococcus aureus, Staphylococcus haemolyticus and Bacillus licheniformis were more prevalent in farms using organic bedding. No obvious associations were found between the main mastitis pathogens and parity or lactation stage. Based on these findings, the dominant pathogens, types of milk, regions, seasons and bedding materials should be considered when designing mastitis prevention and control programs at large Chinese dairy farms.

9.
Ann Palliat Med ; 9(4): 1879-1885, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32576012

RESUMO

BACKGROUND: Triage decision making is crucial for patients arrived at emergency room as it influences the clinical outcome. Appropriate standard to discriminate patients into different category is challenging. We introduced a suitable emergency preview triage pain model for quality management methods through the pain assessment module. METHODS: The pain quality control and management of emergency pre-examination triage were realized by designing the pain assessment module of the intelligent triage system. Clinical nurses completed the assessment of pain score and PQRST pain content through the pain tool in the pain assessment module of the intelligent triage system. The computer system would automatically assess the triage category according to the pain score. Triage nurses determined the priority of emergency care based on the type of care. RESULTS: The pain quality control target monitoring reached 95% of the target management value in ten months. There were two months when the target management value was not reached 95%, June 2019 (94.28%), and December 2019 (94.28%), respectively, slightly lower than the target management value (95%). CONCLUSIONS: The application of the pain quality control management mode of emergency preexamination triage unifies the standards of emergency pre-examination triage pain, standardizes the pain assessment and improves the level of emergency pre-examination triage pain management.

10.
Sci Total Environ ; 740: 140185, 2020 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-32563887

RESUMO

Seawater dissolved organic carbon (DOC) in seagrass meadows is gaining attention for its role in carbon sequestration. Abundant refractory compounds in DOC are exported by seagrass meadows to the deep sea, thereby contributing to long-term carbon drawdown. DOC lability and bacterioplankton communities are key determining factors in this carbon sequestration process, and it has been hypothesized that these may be affected by nutrient loading - however, scientific evidence is so far weak. Here, we studied the response of DOC composition and bacterioplankton communities to nutrient loading in seagrass meadows of the South China Sea. We found that increasing nutrient loads enhanced nitrogen and phosphorus concentrations in DOC, which promoted algae blooms (i.e. epiphyte, phytoplankton and macroalgae) in seagrass meadows, and presumably increased the lability of DOC and its bioavailability to microbes. Also, the relative abundance of K-strategist bacterioplankton communities with the potential to degrade refractory compounds (Acidimicrobiia, Verrucomicrobiales and Micrococcales) increased in the seagrass meadows exposed to high nutrient loads. These results suggest that high nutrient loading can enhance labile DOC composition, and thus increase refractory DOC remineralization rate, thereby weakening the DOC contribution potential of seagrass meadows to long-term carbon sequestration.


Assuntos
Carbono , Ecossistema , China , Sedimentos Geológicos , Nutrientes
11.
J Environ Manage ; 269: 110758, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32560988

RESUMO

Membrane separation technology is recognized as a competitive approach to remove Pb2+ from water system due to its high efficiency and low operating cost. In present study, a simple and facile approach was developed to fabricate covalent organic framework (COF) modified PVDF ultrafiltration membranes with comprehensive antifouling property and superior Pb2+ removal ability. Herein, COF was synthesised in a homogenous PVDF/DMAc solution to fabricate hydrophilic COF modified PVDF ultrafiltration membranes with the Pb2+ removal property. The filtration test demonstrated that the COF modified PVDF ultrafiltration membranes exhibited excellent antifouling property and high water flux. Moreover, the membranes showed remarkable potential for treating Pb2+-containing water. The removal efficiency was determined at 92.4%, and its removal efficiency was 87.5% at the fourth treatment cycle with Pb2+-containing water. The present work provides a valuable platform for further development of efficient composite membranes for the treatment of Pb2+-containing water.


Assuntos
Estruturas Metalorgânicas , Ultrafiltração , Chumbo , Membranas Artificiais , Polivinil
12.
Mol Med Rep ; 22(1): 117-126, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32377732

RESUMO

Acute liver failure (ALF) is a fatal liver disease characterized by severe hepatocyte destruction. MicroRNAs (miRNAs/miRs) have been reported to serve a key role in a number of liver diseases. Therefore, the aim of the present study was to investigate the role and underlying mechanism of miR­214 in ALF. ALF murine and hepatocyte models were established using D­galactosamine (D­GalN) and lipopolysaccharide (LPS) or D­GalN + tumor necrosis factor (TNF)­α, respectively. The expression levels of miR­214 and Bax were detected by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and/or western blotting. Furthermore, an automatic biochemical analyzer was used to measure the levels of aspartate aminotransferase (AST) or alanine aminotransferase (ALT). The levels of TNF­α and interleukin (IL)­6 were detected by ELISA and RT­qPCR. In addition, TUNEL staining and flow cytometry were used to analyze cell apoptosis, and the protein expression of caspase­3 was determined by western blotting. It was identified that the levels of AST and ALT were increased and that hepatocyte apoptosis was enhanced in the D­GalN/LPS­stimulated group compared with the control. Furthermore, higher expression of caspase­3 was observed in the D­GalN/LPS­stimulated group. In addition, it was demonstrated that miR­214 was downregulated, while Bax was upregulated in D­GalN/LPS­stimulated mice and D­GalN/TNF­α­stimulated BNLCL2 cells. Moreover, in D­GalN/TNF­α­stimulated BNLCL2 cells, miR­214 overexpression suppressed apoptosis and decreased TNF­α and IL­6 levels, and these effects were reversed by the Bax plasmid. It was also identified that overexpression of miR­214 significantly decreased Bax mRNA and protein expression levels in vitro. Collectively, the present results suggested that miR­214 inhibited hepatocyte apoptosis during ALF development via targeting Bax, thus indicating that miR­214 may be a potential target for ALF treatment.

13.
Artigo em Inglês | MEDLINE | ID: mdl-32401477

RESUMO

A high-quality precursor solution is essential for the fabrication of hybrid perovskite solar cells. This article reports a simple and efficient method for preparing a high-quality concentrated solution of methylammonium triiodoplumbate (MAPbI3) in N,N-dimethylformamide (DMF) by using MAPbI3 crystals instead of conventional lead iodine and methylammonium iodine blend. The MAPbI3 concentration of the precursor solution is easily and accurately adjusted from 0 up to 1.64 M. An investigation of the dissolution process of the MAPbI3 crystals reveals that the concentrated solution of MAPbI3 in DMF is metastable, and the transition from the concentrated solution to solvated intermediate MAPbI3·DMF determines the solubility of MAPbI3 in DMF. The high purity and precise stoichiometric ratio of the crystals eliminate the possible impurities that initialize the transition to MAPbI3·DMF and consequently suppress the transition and increase the stability of the concentrated solution. MAPbI3 films with different thicknesses up to 800 nm are prepared with the conventional film fabrication technique, and the highest power conversion efficiency of 20.7% is achieved on corresponding solar cells. This newly developed method for preparing a concentrated precursor solution can be easily combined with other fabrication techniques for further development of industrial-scale manufacture of solar cells.

14.
Hum Mol Genet ; 29(9): 1547-1567, 2020 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-32338760

RESUMO

Dominant mutations in the mitochondrial paralogs coiled-helix-coiled-helix (CHCHD) domain 2 (C2) and CHCHD10 (C10) were recently identified as causing Parkinson's disease and amyotrophic lateral sclerosis/frontotemporal dementia/myopathy, respectively. The mechanism by which they disrupt mitochondrial cristae, however, has been uncertain. Using the first C2/C10 double knockout (DKO) mice, we report that C10 pathogenesis and the normal function of C2/C10 are intimately linked. Similar to patients with C10 mutations, we found that C2/C10 DKO mice have disrupted mitochondrial cristae, because of cleavage of the mitochondrial-shaping protein long form of OPA1 (L-OPA1) by the stress-induced peptidase OMA1. OMA1 was found to be activated similarly in affected tissues of mutant C10 knock-in (KI) mice, demonstrating that L-OPA1 cleavage is a novel mechanism for cristae abnormalities because of both C10 mutation and C2/C10 loss. Using OMA1 activation as a functional assay, we found that C2 and C10 are partially functionally redundant, and some but not all disease-causing mutations have retained activity. Finally, C2/C10 DKO mice partially phenocopied mutant C10 KI mice with the development of cardiomyopathy and activation of the integrated mitochondrial integrated stress response in affected tissues, tying mutant C10 pathogenesis to C2/C10 function.

15.
Front Microbiol ; 11: 161, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32194512

RESUMO

Rhizosphere bacterial community structures and their determining drivers have been studied in a variety of marine and freshwater ecosystems for a range of plant species. However, there is still limited information about the influence of habitat on microbial communities in seagrass beds. This study aimed to determine which factors (habitat and plant species) have crucial roles on the rhizospheric bacteria associated with two tropical seagrass species (Thalassia hemprichii and Enhalus acoroides) that are dominant at Xincun Bay and Tanmen Harbor in Hainan Island, South China. Using Illumina HiSeq sequencing, we observed substantial differences in the bacterial richness, diversity, and relative abundances of taxa between the two habitats, which were characterized differently in sediment type and nutrient status. Rhizospheric bacteria from sandy sediment at the eutrophic Xincun Bay were dominated by Desulfobacteraceae and Helicobacteraceae, which are primarily involved in sulfate cycling, whereas rhizosphere microbes from the reef flat at oligotrophic Tanmen Harbor were dominated by Vibrionaceae and Woeseiaceae, which may play important roles in nitrogen and carbon fixing. Additionally, we speculated that host-specific effects of these two seagrass species may be covered under nutrient-rich conditions and in mixed community patches, emphasizing the importance of the nutrient status of the sediment and vegetation composition of the patches. In addition, our study confirmed that Proteobacteria was more adapted to the rhizosphere environment than to low-carbon conditions that occurred in bulk sediment, which was primarily dominated by well-known fermentative bacteria in the phylum Firmicutes.

17.
Mar Pollut Bull ; 150: 110808, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31910532

RESUMO

Heavy metal (Cr, Mn, Fe, Ni, Cu, Zn, Cd, Pb) concentrations in surface sediment porewater and their benthic fluxes were investigated in Daya Bay, South China, to study their accumulation and transfer at the sediment-water interface, as well as the impact of human activities on heavy metals. Heavy metals in porewater displayed different patterns in three partitions (top, center and inlet), which was mainly attributed to the difference in the biogeochemical conditions, hydrodynamic force inner the bay and the human activities along the bay. Ecological risk assessment results showed that heavy metals in porewater dramatically exceeded the background values. The average release of heavy metals from sediment were (6.1 ± 3.3) × 104-(2.7 ± 1.6) × 108 g a-1 in the bay, so they had potential risks to the water environment, and sediment should be paid more attention to as the endogenesis of contamination.


Assuntos
Monitoramento Ambiental , Metais Pesados/análise , Poluentes Químicos da Água , Baías , China , Sedimentos Geológicos , Medição de Risco
18.
Neurosci Bull ; 36(3): 202-216, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31444652

RESUMO

Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2C glycoprotein (SAD-RV(ΔG)-N2C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized. We compared these features in different mouse brain regions for SAD-RV-N2C(G) and two other widely-used retrograde tracers, SAD-RV(ΔG)-B19(G) and rAAV2-retro. We found that SAD-RV(ΔG)-N2C(G) enhanced the infection efficiency of long-projecting neurons by ~10 times but with very similar neuro-tropism, compared with SAD-RV(ΔG)-B19(G). On the other hand, SAD-RV(ΔG)-N2C(G) had an infection efficiency comparable with rAAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of long-projecting neuronal populations. These results demonstrate that SAD-RV(ΔG)-N2C(G) can serve as an effective retrograde vector for studying neuronal circuits.


Assuntos
Giro Denteado , Vetores Genéticos , Glicoproteínas , Rede Nervosa , Técnicas de Rastreamento Neuroanatômico , Vírus da Raiva , Área Tegmentar Ventral , Proteínas Virais , Animais , Linhagem Celular , Cricetinae , Masculino , Camundongos , Camundongos Endogâmicos C57BL
19.
J Cell Biol ; 219(1)2020 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-31719114

RESUMO

Quantification of stable isotope tracers after metabolic labeling provides a snapshot of the dynamic state of living cells and tissue. A form of imaging mass spectrometry quantifies isotope ratios with a lateral resolution <50 nm, using a methodology that we refer to as multi-isotope imaging mass spectrometry (MIMS). Despite lateral resolution exceeding diffraction-limited light microscopy, lack of contrast has largely limited use of MIMS to large or specialized subcellular structures, such as the nucleus and stereocilia. In this study, we repurpose the engineered peroxidase APEX2 as the first genetically encoded marker for MIMS. Coupling APEX2 labeling of lysosomes and metabolic labeling of protein, we identify that individual lysosomes exhibit substantial heterogeneity in protein age, which is lost in iPSC-derived neurons lacking the lysosomal protein progranulin. This study expands the practical use of MIMS for cell biology by enabling measurements of metabolic function from stable isotope labeling within individual organelles in situ.


Assuntos
DNA Liase (Sítios Apurínicos ou Apirimidínicos)/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Marcação por Isótopo/métodos , Lisossomos/metabolismo , Espectrometria de Massas/métodos , Neurônios/metabolismo , Organelas/metabolismo , DNA Liase (Sítios Apurínicos ou Apirimidínicos)/análise , Células HeLa , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Neurônios/citologia , Proteólise
20.
Genes (Basel) ; 10(12)2019 11 28.
Artigo em Inglês | MEDLINE | ID: mdl-31795257

RESUMO

The domain of unknown function (DUF) superfamily encodes proteins of unknown functions in plants. Among them, DUF668 family members in plants possess a 29 amino-acid conserved domain, and this family has not been described previously. Here, we report this plant-specific novel DUF668 gene family containing 12 OsDUF668 genes in rice (Oryza sativa) and 91 DUF668s for the other seven plant species. In our study, DUF668 genes were present in both dicot and monocot plants, indicating that DUF668 is a conserved gene family that originated by predating the dicot-monocot divergence. Based on the gene structure and motif composition, the DUF668 family consists of two distinct clades, I and II in the phylogenetic tree. Remarkably, OsDUF668 genes clustered on the chromosomes merely show close phylogenetic relationships, suggesting that gene duplications or collinearity seldom happened. Cis-elements prediction display that over 80% of DUF668s contain phytohormone and light responsiveness factors. Further comprehensive experimental analyses of the OsDUF668 family are implemented in 22 different tissues, five hormone treatments, seven environmental factor stresses, and two pathogen-defense related stresses. The OsDUF668 genes express ubiquitously in analyzed rice tissues, and seven genes show tissue-specific high expression profiles. All OsDUF668s respond to drought, and some of Avr9/Cf-9 rapidly elicited genes resist to salt, wound, and rice blast with rapidly altered expression patterns. These findings imply that OsDUF668 is essential for drought-enduring and plant defense. Together, our results bring the important role of the DUF668 gene family in rice development and fitness to the fore.


Assuntos
Perfilação da Expressão Gênica/métodos , Família Multigênica , Oryza/crescimento & desenvolvimento , Estresse Fisiológico , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Secas , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Especificidade de Órgãos , Oryza/genética , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética
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