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Xylem-associated fungus can secrete many secondary metabolites to help Aquilaria trees resist various stresses and play a crucial role in facilitating agarwood formation. However, the dynamics of endophytic fungi in Aquilaria sinensis xylem after artificial induction have not been fully elaborated. Endophytic fungi communities and xylem physio-biochemical properties were examined before and after induction with an inorganic salt solution, including four different times (pre-induction (0M), the third (3M), sixth (6M) and ninth (9M) month after induction treatment). The relationships between fungal diversity and physio-biochemical indices were evaluated. The results showed that superoxide dismutase (SOD) and peroxidase (POD) activities, malondialdehyde (MDA) and soluble sugar content first increased and then decreased with induction time, while starch was heavily consumed after induction treatment. Endophytic fungal diversity was significantly lower after induction treatment than before, but the species richness was promoted. Fungal ß-diversity was also clustered into four groups according to different times. Core species shifted from rare to dominant taxa with induction time, and growing species interactions in the network indicate a gradual complication of fungal community structure. Endophytic fungi diversity and potential functions were closely related to physicochemical indices that had less effect on the relative abundance of the dominant species. These findings help assess the regulatory mechanisms of microorganisms that expedite agarwood formation after artificial induction.
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Au@Ag core-shell composites were successfully fabricated on urchin-like covalent organic frameworks (COFs), providing a platform with numerous hot spots for the detection of two categories of emerging contaminants: sulfonamide antibiotics and nanoplastics, using surface-enhanced Raman spectroscopy (SERS). Au seeds (â¼10 nm) were generated on the COFs, leveraging the reducing properties of the vinyl and imino groups within the framework. This ensured the growth of dense and uniformly distributed Ag nanoparticles. The COFs exceptionally large surface area (2324 m2 g-1) and high adsorption capacity, significantly contributed to the enrichment and detection of trace pollutants. As a result, using a portable Raman spectrometer, limits of detection of 0.008 µmol L-1 for sulfamethoxazole and 0.029 mg L-1 for polystyrene nanoplastics were achieved.
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The leaf economics spectrum (LES) describes the covariation of traits relevant for carbon and nutrient economy in different plant species. However, much less is known about the correlation of LES with leaf water economy, not only because some woody species do not follow the rules, but also because they are rarely tested on the widespread, non-native, fast-growing trees. We hypothesized that fast-growing exotic species that spread on the fast side of the LES coordinate their water-use strategies (WUS) to maintain rapid growth, and that the pattern of coordination differs between evergreen and deciduous forests. Using 4 exotic and 4 native species from evergreen and deciduous broadleaf forests in China, we measured 17 traits of LES and WUS and analyzed their functional roles in different species groups. Our results suggest that LES plays a more important role in the coexistence of species within a community, while WUS contributes more to the distribution of species across different regions. The multidimensional coordination of LES and WUS could better explain the growth and distribution of different plant species and shed light on the coexistence of species from different forest types, especially fast-growing woody exotics.
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Bosques , Especies Introducidas , Hojas de la Planta , Árboles , China , AguaRESUMEN
Quercus gilva, an evergreen tree species in Quercus section Cyclobalanopsis, is an ecologically and economically valuable species in subtropical regions of East Asia. Predicting the impact of climate change on potential distribution of Q. gilva can provide a scientific basis for the conservation and utilization of its genetic resources, as well as for afforestation. In this study, 74 distribution records of Q. gilva and nine climate variables were obtained after data collection and processing. Current climate data downloaded from WorldClim and future climate data predicted by four future climate scenarios (2040s SSP1-2.6, 2040s SSP5-8.5, 2060s SSP1-2.6, and 2060s SSP5-8.5) mainly based on greenhouse gases emissions of distribution sites were used in MaxEnt model with optimized parameters to predict distribution dynamics of Q. gilva and its response to climate change. The results showed that the predicted current distribution was consistent with natural distribution of Q. gilva, which was mainly located in Hunan, Jiangxi, Zhejiang, Fujian, Guizhou, and Taiwan provinces of China, as well as Japan and Jeju Island of South Korea. Under current climate conditions, precipitation factors played a more significant role than temperature factors on distribution of Q. gilva, and precipitation of driest quarter (BIO17) is the most important restriction factor for its current distribution (contribution rate of 57.35%). Under future climate conditions, mean temperature of driest quarter (BIO9) was the essential climate factor affecting future change in potential distribution of Q. gilva. As the degree of climatic anomaly increased in the future, the total area of predicted distribution of Q. gilva showed a shrinking trend (decreased by 12.24%-45.21%) and Q. gilva would migrate to high altitudes and latitudes. The research results illustrated potential distribution range and suitable climate conditions of Q. gilva, which can provide essential theoretical references for the conservation, development, and utilization of Q. gilva and other related species.
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Gases de Efecto Invernadero , Quercus , Cambio Climático , China , Taiwán , EcosistemaRESUMEN
Trehalose-6-phosphate phosphatase (TPP) is a pivotal enzyme in trehalose biosynthesis which plays an essential role in plant development and in the abiotic stress response. However, little is currently known about TPPs in groundnut. In the present study, a total of 16 AhTPP genes were identified, and can be divided into three phylogenetic subgroups. AhTPP members within the same subgroups generally displayed similar exon-intron structures and conserved motifs. Gene collinearity analysis revealed that segmental duplication was the primary factor driving the expansion of the AhTPP family. An analysis of the upstream promoter region of AhTPPs revealed eight hormone- and four stress-related responsive cis-elements. Transcriptomic analysis indicated high expression levels of AhTPP genes in roots or flowers, while RT-qPCR analysis showed upregulation of the six tested genes under different abiotic stresses, suggesting that AhTPPs play roles in growth, development, and response to various abiotic stresses. Subcellular localization analysis showed that AhTPP1A and AhTPP5A were likely located in both the cytoplasm and the nucleus. To further confirm their functions, the genes AhTPP1A and AhTPP5A were individually integrated into yeast expression vectors. Subsequent experiments demonstrated that yeast cells overexpressing these genes displayed increased tolerance to osmotic and salt stress compared to the control group. This study will not only lay the foundation for further study of AhTPP gene functions, but will also provide valuable gene resources for improving abiotic stress tolerance in groundnut and other crops.
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Introduction: Peanut (Arachis hypogaea L.), also called groundnut is an important oil and cash crop grown widely in the world. The annual global production of groundnuts has increased to approximately 50 million tons, which provides a rich source of vegetable oils and proteins for humans. Low temperature (non-freezing) is one of the major factors restricting peanut growth, yield, and geographic distribution. Since the complexity of cold-resistance trait, the molecular mechanism of cold tolerance and related gene networks were largely unknown in peanut. Methods: In this study, comparative transcriptomic analysis of two peanut cultivars (SLH vs. ZH12) with differential cold tolerance under low temperature (10°C) was performed using Oxford Nanopore Technology (ONT) platform. Results and discussion: As a result, we identified 8,949 novel gene loci and 95,291 new/novel isoforms compared with the reference database. More differentially expressed genes (DEGs) were discovered in cold-sensitive cultivar (ZH12) than cold-tolerant cultivar (SLH), while more alternative splicing events were found in SLH compared to ZH12. Gene Ontology (GO) analyses of the common DEGs showed that the "response to stress", "chloroplast part", and "transcription factor activity" were the most enriched GO terms, indicating that photosynthesis process and transcription factors play crucial roles in cold stress response in peanut. We also detected a total of 708 differential alternative splicing genes (DASGs) under cold stress compared to normal condition. Intron retention (IR) and exon skipping (ES) were the most prevalent alternative splicing (AS) events. In total, 4,993 transcription factors and 292 splicing factors were detected, many of them had differential expression levels and/or underwent AS events in response to cold stress. Overexpression of two candidate genes (encoding trehalose-6-phosphatephosphatases, AhTPPs) in yeast improves cold tolerance. This study not only provides valuable resources for the study of cold resistance in peanut but also lay a foundation for genetic modification of cold regulators to enhance stress tolerance in crops.
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Busulfan is an antineoplastic, which is always accompanied with the abnormal of spermatogonia self-renewal and differentiation. It has been demonstrated that the omega-3 polyunsaturated fatty acids (PUFAs) benefits mature spermatozoa. However, whether omega-3 can protect endogenous spermatogonia and the detailed mechanisms are still unclear. Evaluate of spermatogenesis function (in vivo) were examined by histopathological analysis, immunofluorescence staining, and western blotting. The levels of lipid metabolites in testicular tissue were determined via liquid chromatography. We investigated the effect of lipid metabolites on Sertoli cells provided paracrine factors to regulate spermatogonia proliferation and differentiation using co-culture system. In our study, we showed that omega-3 PUFAs significantly improved the process of sperm production and elevated the quantity of both undifferentiated Lin28+ spermatogonia and differentiated c-kit+ spermatogonia in a mouse model where spermatogenic function was disrupted by busulfan. Mass spectrometry revealed an increase in the levels of several omega-3 metabolites in the testes of mice fed with omega-3 PUFAs. The eicosapentaenoic acid metabolite 12-hydroxyeicosapentaenoic acid (12-HEPE) up-regulated bone morphogenic protein 4 (BMP4) expression through GPR120-ERK1/2 pathway activation in Sertoli cells and restored spermatogonia proliferation and differentiation. Our study provides evidence that omega-3 PUFAs metabolite 12-HEPE effectively protects spermatogonia and reveals that GPR120 might be a tractable pharmacological target for fertility in men received chemotherapy or severe spermatogenesis dysfunction.
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Busulfano , Semen , Humanos , Masculino , Ratones , Animales , Busulfano/farmacología , Busulfano/metabolismo , Espermatogénesis/fisiología , Espermatogonias , Espermatozoides , Testículo/metabolismoRESUMEN
BACKGROUND: Glycosylation, catalyzed by UDP-glycosyltransferase (UGT), was important for enhancing solubility, bioactivity, and diversity of flavonoids. Peanut (Arachis hypogaea L.) is an important oilseed and cash crop worldwide. In addition to provide high quality of edible oils and proteins, peanut seeds contain a rich source of flavonoid glycosides that benefit human health. However, information of UGT gene family was quite limited in peanut. RESULTS: In present study, a total of 267 AhUGTs clustered into 15 phylogenetic groups were identified in peanut genome. Group I has greatly expanded to contain the largest number of AhUGT genes. Segmental duplication was the major driving force for AhUGT gene family expansion. Transcriptomic analysis of gene expression profiles in various tissues and under different abiotic stress treatments indicated AhUGTs were involved in peanut growth and abiotic stress response. AhUGT75A (UGT73CG33), located in mitochondria, was characterized as a flavonoid 7-O-UGT by in vitro enzyme assays. The transcript level of AhUGT75A was strongly induced by abiotic stress. Overexpression of AhUGT75A resulted in accumulating less amount of malondialdehyde (MDA) and superoxide, and enhancing tolerance against drought and/or salt stress in transgenic Arabidopsis. These results indicated AhUGT75A played important roles in conferring abiotic stress tolerance through reactive oxygen species scavenging. CONCLUSIONS: Our research only not provides valuable information for functional characterization of UGTs in peanut, but also gives new insights into potential applications in breeding new cultivars with both desirable stress tolerance and health benefits.
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Arabidopsis , Arachis , Humanos , Arachis/genética , Glicosiltransferasas/genética , Filogenia , Flavonoides , Fitomejoramiento , Estrés Fisiológico/genética , Uridina DifosfatoRESUMEN
While SARS-CoV-2 is generally under control, the question of variants and infections still persists. Fundamental information on how the virus interacts with inanimate surfaces commonly found in our daily life and when in contact with the skin will be helpful in developing strategies to inhibit the spread of the virus. Here in, a critically important review of current understanding of the interaction between virus and surface is summarized from chemistry point-of-view. The Derjaguin-Landau-Verwey-Overbeek and extended Derjaguin-Landau-Verwey-Overbeek theories to model virus attachments on surfaces are introduced, along with the interaction type and strength, and quantification of each component. The virus survival and transfer are affected by a combination of biological, physical, and chemical parameters, as well as environmental parameters. The surface properties for virus and virus survival on typical surfaces such as metals, plastics, and glass are summarized. Attention is also paid to the transfer of virus to/from surfaces and skin. Typical virus disinfection strategies utilizing heat, light, chemicals, and ozone are discussed together with their disinfection mechanism. In the last section, design principles for virus repelling surface chemistry such as surperhydrophobic or surperhydrophilic surfaces are also introduced, to demonstrate how the integration of surface property control and advanced material fabrication can lead to the development of functional surfaces for mitigating the effect of viral infection upon contact.
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Desinfección , Propiedades de SuperficieRESUMEN
BACKGROUND: Polycystic ovary syndrome (PCOS) is a common reproductive endocrine disorder that frequently exhibits low-grade inflammation, pro-oxidant activity, and gut dysbiosis. PCOS has become one of the leading causes of female infertility worldwide. Recently, omega-3 polyunsaturated fatty acids (PUFAs) have been proven to benefit metabolic disorders in PCOS patients. However, its roles in the regulation of metabolic and endocrinal balances in PCOS pathophysiology are not clear. In the present study, we aimed to explore how omega-3 PUFAs alleviate ovarian dysfunction and insulin resistance in mice with dehydroepiandrosterone (DHEA)-induced PCOS by modulating the gut microbiota. METHODS: We induced PCOS in female mice by injecting them with DHEA and then treated them with omega-3 PUFAs. 16S ribosomal DNA (rDNA) amplicon sequencing, fecal microbiota transplantation (FMT) and antibiotic treatment were used to evaluate the role of microbiota in the regulation of ovarian functions and insulin resistance (IR) by omega-3 PUFAs. To further investigate the mechanism of gut microbiota on omega-3-mediated ovarian and metabolic protective effects, inflammatory and oxidative stress markers in ovaries and thermogenic markers in subcutaneous and brown adipose tissues were investigated. RESULTS: We found that oral supplementation with omega-3 PUFAs ameliorates the PCOS phenotype. 16S rDNA analysis revealed that omega-3 PUFA treatment increased the abundance of beneficial bacteria in the gut, thereby alleviating DHEA-induced gut dysbiosis. Antibiotic treatment and FMT experiments further demonstrated that the mechanisms underlying omega-3 benefits likely involve direct effects on the ovary to inhibit inflammatory cytokines such as IL-1ß, TNF-α and IL-18. In addition, the gut microbiota played a key role in the improvement of adipose tissue morphology and function by decreasing multilocular cells and thermogenic markers such as Ucp1, Pgc1a, Cited and Cox8b within the subcutaneous adipose tissues. CONCLUSION: These findings indicate that omega-3 PUFAs ameliorate androgen-induced gut microbiota dysbiosis. The gut microbiota plays a key role in the regulation of omega-3-mediated IR protective effects in polycystic ovary syndrome mice. Moreover, omega-3 PUFA-regulated improvements in the ovarian dysfunction associated with PCOS likely involve direct effects on the ovary to inhibit inflammation. Our findings suggest that omega-3 supplementation may be a promising therapeutic approach for the treatment of PCOS by modulating gut microbiota and alleviating ovarian dysfunction and insulin resistance.
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Suplementos Dietéticos , Ácidos Grasos Omega-3 , Microbioma Gastrointestinal , Síndrome del Ovario Poliquístico , Animales , Femenino , Ratones , Deshidroepiandrosterona/toxicidad , Microbioma Gastrointestinal/fisiología , Resistencia a la Insulina , Síndrome del Ovario Poliquístico/inducido químicamente , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Síndrome del Ovario Poliquístico/fisiopatología , Ácidos Grasos Omega-3/uso terapéuticoRESUMEN
Exploration of the remobilization mechanism of trace metals in estuarine sediments remain challenging because of dynamic hydrochemical conditions. This study integrated a chemical sequential extraction procedure (BCR), the diffusive gradient in thin films (DGT) and high-resolution dialysis techniques, and Visual MINTEQ ver.3.1 to identify the seasonal mobilization characteristics of sediment Zn within a tidal reach, South China. The mobility of sediment Zn based on the BCR procedure contradicted the results of DGT analysis. In summer, reductive dissolution of Fe/Mn oxides was the key driver of sediment Zn remobilization; during winter, cation exchange reactions facilitated the mobilization of Zn in the brackish water zone. The time-dependence ratios of DGT-labile Zn and dissolved Zn concentrations (mean: 0.34-0.81) indicated the sediment solid phase had partially sustained capacity to resupply Zn to the porewater in both seasons. Sediments generally functioned as a source of Zn in the freshwater zone with organically complexed Zn being diffusively released into the water column at rates of 0.3-15.5 µg·m-2·d-1. In the brackish water zone, the dominant Zn species were transformed into free Zn ions and Zn-inorganic complexes and migrated into sediment, with respective influxes of 18.9-70.7 µg·m-2·d-1 and 18.9-68.3 µg·m-2·d-1, which shifted to a sink of Zn.
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A new method was developed for the coagulation of arsenic with serpentine and Fe(II). Excellent removal efficiency (>99%) and satisfactory stability of the sediments were accomplished for As(V) and As(III). A mechanism study showed that hydroxyls generated by the surface hydrolysis of serpentine mediated active iron hydroxides for arsenic adsorption, while the Fe-As and Mg-As chemical interaction contributed to the arsenic stabilization.
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MAIN CONCLUSION: We developed a more realistic modeling framework by integrating stem photosynthesis into the canopy carbon assimilation model to compare the photosynthetic productivity between the stem and leaf of Eucalyptus urophylla plantations. Stems of Eucalyptus species with smooth outer bark have photosynthetic green tissue that can recycle internal stem CO2. However, the potential contribution of stem photosynthesis to forest productivity has not previously been adequately quantified, and we also do not know how it compares to leaf photosynthetic productivity. To assist in addressing this knowledge gap, we conducted field surveys in Eucalyptus urophylla plantations of different ages and developed a more realistic modeling framework by integrating stem photosynthesis into the existing canopy carbon assimilation model. We calculated the proportion of tree stems shaded by neighboring tree trunks based on Poisson spatial point process. Under the stand density of 2000 trees per hectare, the light absorption area of tree trunks of 2-year-old and 7-year-old E. urophylla plantations were 0.11 (± 0.15) and 0.35 (± 0.12) m2 stem m-2 land, the stem photosynthetic productivity (GPPstem) was 0.72 (± 0.45) and 1.81 (± 1.12) mol C m-2 month-1, and the ratios of GPPstem to leaf photosynthetic productivity (GPPleaf) were 5.10 and 8.17% for 2- and 7-year-old plantations, respectively. Overall, this study presents the feasibility of incorporating stem photosynthesis into the productivity prediction of E. urophylla plantations by developing the stem light absorption model.
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Eucalyptus , Fotosíntesis , Árboles , Hojas de la Planta , CarbonoRESUMEN
Cancer cells prefer glycolysis to support their proliferation. Our previous studies have shown that the long palate, lung, and nasal epithelial cell clone 1 (LPLUNC1) can upregulate prohibitin 1 (PHB1) expression to inhibit the proliferation of nasopharyngeal carcinoma (NPC) cells. Given that PHB1 is an important regulator of cell energy metabolism, we explored whether and how LPLUNC1 regulated glucose glycolysis in NPC cells. LPLUNC1 or PHB1 overexpression decreased glycolysis and increased oxidative phosphorylation (OXPHOS)-related protein expression in NPC cells, promoting phosphorylated PHB1 nuclear translocation through 14-3-3σ. LPLUNC1 overexpression also increased p53 but decreased c-Myc expression in NPC cells, which were crucial for the decrease in glycolysis and increase in OXPHOS-related protein expression induced by LPLUNC1 overexpression. Finally, we found that treatment with all-trans retinoic acid (ATRA) reduced the viability and clonogenicity of NPC cells, decreased glycolysis, and increased OXPHOS-related protein expression by enhancing LPLUNC1 expression in NPC cells. Therefore, the LPLUNC1-PHB1-p53/c-Myc axis decreased glycolysis in NPC cells, and ATRA upregulated LPLUNC1 expression, ATRA maybe a promising drug for the treatment of NPC.
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Neoplasias Nasofaríngeas , Proteína p53 Supresora de Tumor , Humanos , Línea Celular Tumoral , Proliferación Celular , Células Epiteliales/patología , Regulación Neoplásica de la Expresión Génica , Glucólisis , Carcinoma Nasofaríngeo/genética , Neoplasias Nasofaríngeas/patología , Tretinoina/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Proteínas de Unión a Ácidos Grasos/metabolismo , Autoantígenos/metabolismoRESUMEN
Flow parameters measurement facilitates the understanding of two-phase flow. Due to the changeable structures of the flow, the prediction of superficial velocity of oil-water two-phase flow in large diameter pipes is still a challenging problem. Therefore, we first conducted a vertical upward oil-water two-phase flow experiment in a 125 mm ID pipe, and obtained the response signals under different flow conditions by a vertical multi-electrode array (VMEA) conductance sensor. Then, new data pre-processing (1D to 2D) techniques and information fusion techniques (network channels) are employed. Moreover, the front-end structure of the network is optimized using a combination of attention block and residual structure, and the middle structure is optimized using inception block; on the other hand, the back-end structure of the original capsule network is innovatively changed so that it can handle both the flow pattern classification and superficial velocity prediction tasks. The dynamic routing algorithm has also been improved to speed up model training. Extensive experiments validate the effectiveness of the improved modules. Finally, we compare the proposed network with its variants and other competing networks. The better performance results show that our multi-task sequence-based CapsNet has great potential for dealing with high-dimensional, time-varying and nonlinear problems in multiphase flow.
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It is highly desirable to produce bread with both acceptable texture and health benefits. In this study, maltohexaose (G6) producing amylase AmyM and its truncation AmyM-TR2 from Corallococcus sp. strain EGB were used to determine their effects to bread quality and starch physicochemical properties. During bread fermentation, AmyM or AmyM-TR2 continuously degraded the starch, resulting in more obvious decrease in relative crystallinity, the ordered structure, pasting viscosities and gelatinization enthalpy of starch than in control. The dough treated with AmyM or AmyM-TR2 increased bread volume and slowly digestible starch content, decreased bread hardness, and extended bread shelf life and as compared with control, and the dough treated with AmyM-TR2 had better improvement effects than AmyM. The volume and slowly digestible starch content of bread from the treatment of AmyM-TR2 increased by 9.74% and 7.56% in normal wheat, 1.42% and 10.28% in waxy wheat as compared with AmyM, respectively. AmyM-TR2 affected the substrate targeting, proximity and structure disruption effects, which contributed to the degradation of more starch than AmyM.
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Pan , Triticum , Almidón , alfa-Amilasas , CerasRESUMEN
Peanut (also called groundnut, Arachis hypogaea L.) seeds are used for producing edible oils and functional foods, and offer a rich source of lipids, proteins and carbohydrates. However, the location of these metabolites has not yet been firmly established. In the present study, the matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI) technique was applied to investigate spatial distribution of lipids and other key components in seeds of three peanut cultivars (ZH9, KQBH, HP). A total of 103 metabolites, including 34 lipid compounds, were putatively identified by MALDI-MSI. The abundance and spatial distribution of glycerolipids (GLs) and glycerophospholipids (GPs) were compared among the three peanut cultivars. All the identified lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE) and phosphatidylcholines (PCs) were distributed mainly in the inner part of seeds. The visualization of phosphatidic acids (PAs) and triacylglycerols (TGs) revealed a dramatic metabolic heterogeneity between the different tissues making up the seed. The non-homogeneous spatial distribution of metabolites appeared to be related to the different functions of particular tissue regions. These results indicated that MALDI-MSI could be useful for investigating the lipids of foodstuffs from a spatial perspective. The present study may contribute to the development of oil crops with higher oil yields, and to improvement of food processing.
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Three dimensional deformable image registration (DIR) is a key enabling technique in building digital neuronal atlases of the brain, which can model the local non-linear deformation between a pair of biomedical images and align the anatomical structures of different samples into one spatial coordinate system. And thus, the DIR is always conducted following a preprocessing of global linear registration to remove the large global deformations. However, imperfect preprocessing may leave some large non-linear deformations that cannot be handled well by existing DIR methods. The recently proposed cascaded registration network gives a primary solution to deal with such large non-linear deformations, but still suffers from loss of image details caused by continuous interpolation (information loss problem). In this article, a progressive image registration strategy based on deep self-calibration is proposed to deal with the large non-linear deformations without causing information loss and introducing additional parameters. More importantly, we also propose a novel hierarchical registration strategy to quickly achieve accurate multi-scale progressive registration. In addition, our method can implicitly and reasonably implement dynamic dataset augmentation. We have evaluated the proposed method on both optical and MRI image datasets with obtaining promising results, which demonstrate the superior performance of the proposed method over several other state-of-the-art approaches for deformable image registration.
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Sertoli cells are essential for spermatogenesis in the testicular seminiferous tubules by forming blood-testis barrier (BTB) and creating a unique microenvironment for spermatogenesis. Many lncRNAs have been reported to participate in spermatogenesis. However, the role of long noncoding RNAs (lncRNAs) in Sertoli cells has rarely been examined. Herein, we found that a high-fat diet (HFD) decreased sperm quality, impaired BTB integrity and resulted in accumulation of saturated fatty acids (SFAs), especially palmitic acid (PA), in mouse testes. PA decreased the expression of tight junction (TJ)-related proteins, increased permeability and decreased transepithelial electrical resistance (TER) in primary Sertoli cells and TM4 cells. Moreover, lncRNA Tug1 was found to be involved in PA-induced BTB disruption by RNA-seq. Tug1 depletion distinctly impaired the TJs of Sertoli cells and overexpression of Tug1 alleviated the disruption of BTB integrity induced by PA. Moreover, Ccl2 was found to be a downstream target of Tug1, and decreased TJ-related protein levels and TER and increased FITC-dextran permeability in vitro. Furthermore, the addition of Ccl2 damaged BTB integrity after overexpression of Tug1 in the presence of PA. Mechanistically, we found that Tug1 could directly bind to EZH2 and regulate H3K27me3 occupancy in the Ccl2 promoter region by RNA immunoprecipitation and chromatin immunoprecipitation assays. Our study revealed an important role of Tug1 in the BTB integrity of Sertoli cells and provided a new view of the role of lncRNAs in male infertility.
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Barrera Hematotesticular/metabolismo , ARN Largo no Codificante/genética , Túbulos Seminíferos/irrigación sanguínea , Células de Sertoli/metabolismo , Espermatogénesis/genética , Uniones Estrechas/genética , Animales , Células Cultivadas , Quimiocina CCL2/biosíntesis , Quimiocina CCL2/genética , Dieta Alta en Grasa , Impedancia Eléctrica , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Histonas/metabolismo , Infertilidad Masculina/genética , Masculino , Ratones , Ratones Endogámicos ICR , Obesidad/patología , Ácido Palmítico/análisis , Análisis de Semen , Espermatogénesis/fisiologíaRESUMEN
Obtaining high-quality sperm is key to improving the success rate of assisted reproductive technology (ART). Although cytokines secreted by cumulus-oocyte complexes (COCs) bind to sperm surface receptors to improve sperm quality, the effects of adding mouse COCs to human tubal fluid (HTF) medium on sperm capacitation have not yet been explored. Eight-week-old ICR mouse COCs were added to HTF medium and crushed to obtain the post-modified HTF medium. Compared with using HTF medium, the fertilisation rate and number of sperm combined with the zona pellucida significantly increased after in vitro capacitation using the post-modified HTF medium (P < 0.01). Proteomic and Western blotting analyses showed that the level of SERPINA5 in sperm increased significantly following in vitro capacitation with the post-modified HTF medium (P < 0.05). Immunohistochemical staining analysis demonstrated that SERPINA5 protein was expressed in mouse cumulus cells. A SERPINA5 antibody was added in the post-modified HTF medium to block the effects of SERPINA5 after in vitro capacitation, which significantly decreased the fertilisation rate and the number of sperm combined with the zona pellucida (P < 0.05). Recombinant mouse SERPINA5 protein (1 ~ 2 µg/ml) was added to HTF medium and the fertilisation rate and the number of sperm combined with the zona pellucida significantly increased (P < 0.01). Moreover, recombinant human SERPINA5 protein (5 µg/ml) was added before human semen freezing. Compared with adding no SERPINA5 protein, the percentage of normal sperm morphology and the intact acrosome significantly increased (P < 0.05). Our study provides a reference method for optimising sperm quality in the process of in vitro capacitation.