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Common wheat (Triticum aestivum) is one of the most widely cultivated and consumed crops globally. In the face of limited arable land and climate changes, it is a great challenge to maintain current and increase future wheat production. Enhancing agronomic traits in wheat by introducing mutations across all three homoeologous copies of each gene has proven to be a difficult task due to its large genome with high repetition. However, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated nuclease (Cas) genome editing technologies offer a powerful means of precisely manipulating the genomes of crop species, thereby opening up new possibilities for biotechnology and breeding. In this review, we first focus on the development and optimization of the current CRISPR-based genome editing tools in wheat, emphasizing recent breakthroughs in precise and multiplex genome editing. We then describe the general procedure of wheat genome editing and highlight different methods to deliver the genome editing reagents into wheat cells. Furthermore, we summarize the recent applications and advancements of CRISPR/Cas technologies for wheat improvement. Lastly, we discuss the remaining challenges specific to wheat genome editing and its future prospects.
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Sistemas CRISPR-Cas , Triticum , Sistemas CRISPR-Cas/genética , Triticum/genética , Genoma de Planta/genética , Melhoramento Vegetal , Edição de Genes/métodosRESUMO
Prime editing is limited by low efficiency in plants. Here, we develop an upgraded engineered plant prime editor in hexaploid wheat, ePPEplus, by introducing a V223A substitution into reverse transcriptase in the ePPEmax* architecture. ePPEplus enhances the efficiency by an average 33.0-fold and 6.4-fold compared to the original PPE and ePPE, respectively. Importantly, a robust multiplex prime editing platform is established for simultaneous editing of four to ten genes in protoplasts and up to eight genes in regenerated wheat plants at frequencies up to 74.5%, thus expanding the applicability of prime editors for stacking of multiple agronomic traits.
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Agricultura , Triticum , Triticum/genética , FenótipoRESUMO
Metasurface polarization optics that consist of 2D array of birefringent nano-antennas have proven remarkable capabilities to generate and manipulate vectorial fields with subwavelength resolution and high efficiency. Integrating this new type of metasurface with the standard vertical cavity surface-emitting laser (VCSEL) platform enables an ultracompact and powerful solution to control both phase and polarization properties of the laser on a chip, which allows to structure a VCSEL into vector beams with on-demand wavefronts. Here, this concept is demonstrated by directly generating versatile vector beams from commercially available VCSELs through on-chip integration of high-index dielectric metasurfaces. Experimentally, the versatility of the approach for the development of vectorial VCSELs are validated by implementing a variety of functionalities, including directional emission of multibeam with specified polarizations, vectorial holographic display, and vector vortex beams generations. Notably, the proposed vectorial VCSELs integrated with a single layer of beam shaping metasurface bypass the requirements of multiple cascaded optical components, and thus have the potential to promote the advancements of ultracompact, lightweight, and scalable vector beams sources, enriching and expanding the applications of VCSELs in optical communications, laser manipulation and processing, information encryption, and quantum optics.
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Polarization response of artificially structured nano-antennas can be exploited to design innovative optical components, also dubbed "vectorial metasurfaces", for the modulation of phase, amplitude, and polarization with subwavelength spatial resolution. Recent efforts in conceiving Jones matrix formalism led to the advancement of vectorial metasurfaces to independently manipulate any arbitrary phase function of orthogonal polarization states. Here, we are taking advantages of this formalism to design and experimentally validate the performance of CMOS compatible Jones matrix metasurfaces monolithically integrated with standard VCSELs for on-chip spin-decoupling and phase shaping. Our approach enables accessing the optical spin states of VCSELs in an ultra-compact way with previously unattainable phase controllability. By exploiting spin states as a new degree of freedom for laser wavefront engineering, our platform is capable of operating and reading-out the spin-momentum of lasers associated with injected spin carriers, which would potentially play a pivotal role for the development of emerging spin-optoelectronic devices.
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Ketamine, a rapid-acting antidepressant drug, has been used to treat major depressive disorder and bipolar disorder (BD). Recent studies have shown that ketamine may increase the potential risk of treatment-induced mania in patients. Ketamine has also been applied to establish animal models of mania. At present, however, the underlying mechanism is still unclear. In the current study, we found that chronic lithium exposure attenuated ketamine-induced mania-like behavior and c-Fos expression in the medial prefrontal cortex (mPFC) of adult male mice. Transcriptome sequencing was performed to determine the effect of lithium administration on the transcriptome of the PFC in ketamine-treated mice, showing inactivation of the phosphoinositide 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway. Pharmacological inhibition of AKT signaling by MK2206 (40 mg/kg), a selective AKT inhibitor, reversed ketamine-induced mania. Furthermore, selective knockdown of AKT via AAV-AKT-shRNA-EGFP in the mPFC also reversed ketamine-induced mania-like behavior. Importantly, pharmacological activation of AKT signaling by SC79 (40 mg/kg), an AKT activator, contributed to mania in low-dose ketamine-treated mice. Inhibition of PI3K signaling by LY294002 (25 mg/kg), a specific PI3K inhibitor, reversed the mania-like behavior in ketamine-treated mice. However, pharmacological inhibition of mammalian target of rapamycin (mTOR) signaling with rapamycin (10 mg/kg), a specific mTOR inhibitor, had no effect on ketamine-induced mania-like behavior. These results suggest that chronic lithium treatment ameliorates ketamine-induced mania-like behavior via the PI3K-AKT signaling pathway, which may be a novel target for the development of BD treatment.
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Transtorno Depressivo Maior , Ketamina , Doenças dos Roedores , Masculino , Camundongos , Animais , Ketamina/toxicidade , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Fosfatidilinositol 3-Quinases/farmacologia , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-akt/farmacologia , Lítio/farmacologia , Mania , Fosfatidilinositol 3-Quinase/genética , Fosfatidilinositol 3-Quinase/metabolismo , Fosfatidilinositol 3-Quinase/farmacologia , RNA Interferente Pequeno , Serina-Treonina Quinases TOR/genética , Transdução de Sinais , Antidepressivos/uso terapêutico , Antidepressivos/farmacologia , Sirolimo/farmacologia , Compostos de Lítio/farmacologia , Mamíferos , Doenças dos Roedores/tratamento farmacológicoRESUMO
The Siberian Traps large igneous province (STLIP) is commonly invoked as the primary driver of global environmental changes that triggered the end-Permian mass extinction (EPME). Here, we explore the contributions of coeval felsic volcanism to end-Permian environmental changes. We report evidence of extreme Cu enrichment in the EPME interval in South China. The enrichment is associated with an increase in the light Cu isotope, melt inclusions rich in copper and sulfides, and Hg concentration spikes. The Cu and Hg elemental and isotopic signatures can be linked to S-rich vapor produced by felsic volcanism. We use these previously unknown geochemical data to estimate volcanic SO2 injections and argue that this volcanism would have produced several degrees of rapid cooling before or coincident with the more protracted global warming. Large-scale eruptions near the South China block synchronous with the EPME strengthen the case that the STLIP may not have been the sole trigger.
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BACKGROUND: The advent of immune checkpoint inhibitors (ICIs) has revolutionized the management of several types of solid cancers, including lung cancer, by boosting the body's natural tumor killing response. However, it is undeniable that only a small proportion of non-small cell lung cancer (NSCLC) patients with epidermal growth factor receptor (EGFR) mutations can achieve long-term responses and benefit from immunotherapy. CASE SUMMARY: Herein, we report the case of a 48-year-old man diagnosed with stage IV lung adenocarcinoma with an EGFR L858R mutation who was administered pembrolizumab monotherapy followed by pemetrexed and achieved a 10-month progression-free survival interval. In this case report, we show that ICIs were effective for our patient with EGFR-mutated NSCLC and discuss the characteristics of patients who can benefit from immunotherapy. CONCLUSION: We suggest that patients with EGFR-mutated NSCLC with high PD-L1 expression (defined as ≥ 25%), the L858R mutation, smoking history, or pemetrexed pretreatment may benefit from immunotherapy.
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Clavicle hook plate is a common implant for treating distal clavicle fracture. Although high bone union rate and good functional outcome have been reported, so have several complications, such as osteolysis and fracture of the acromion, loss reduction, hook impingement, and rotator cuff tear. Peri-implant fracture over the medial side of the hook plate is a rare complication. Sporadic cases have been reported, and most of them have had no history of trauma. Between June 2015 and August 2018, 7 patients treated for distal clavicle fracture with a 3.5-mm locking compression hook plate with no history of trauma experienced peri-implant fracture of the medial clavicle. This complication occurred at a mean of 29 days. The incidence rate was 9.8%. Peri-implant fracture following hook plate fixation for distal clavicle fracture was not rare. Small hook angle, prolonged retention of the implant, an eccentric medial screw, high plate screw density, and small clavicle diameter may be risk factors for peri-implant fracture. Regarding treatment, 2 patients chose fracture revision with a distal clavicle locking plate and 5 patients chose conservative treatment. All patients achieved bone union at fracture sites. Surgical and conservative management of peri-implant fracture can achieve good functional outcome. [Orthopedics. 2020;43(5);e359-e363.].
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Placas Ósseas/efeitos adversos , Clavícula/lesões , Fixação Interna de Fraturas/efeitos adversos , Fraturas Ósseas/cirurgia , Fraturas Periprotéticas/etiologia , Parafusos Ósseos , Clavícula/cirurgia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fraturas Periprotéticas/cirurgia , Reoperação , Estudos Retrospectivos , Fatores de Risco , Resultado do Tratamento , Adulto JovemRESUMO
Vertical cavity surface-emitting lasers (VCSELs) have made indispensable contributions to the development of modern optoelectronic technologies. However, arbitrary beam shaping of VCSELs within a compact system has remained inaccessible until now. The emerging ultra-thin flat optical structures, namely metasurfaces, offer a powerful technique to manipulate electromagnetic fields with subwavelength spatial resolution. Here, we show that the monolithic integration of dielectric metasurfaces with VCSELs enables remarkable arbitrary control of the laser beam profiles, including self-collimation, Bessel and Vortex lasers, with high efficiency. Such wafer-level integration of metasurface through VCSEL-compatible technology simplifies the assembling process and preserves the high performance of the VCSELs. We envision that our approach can be implemented in various wide-field applications, such as optical fibre communications, laser printing, smartphones, optical sensing, face recognition, directional displays and ultra-compact light detection and ranging (LiDAR).
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BACKGROUND: Studies have shown that HSP20 (heat-shock protein 20) genes play important roles in regulating plant growth, development, and stress response. However, the grape HSP20 gene family has not been well studied. RESULTS: A total of 48 VvHSP20 genes were identified from the grape genome, which were divided into 11 subfamilies (CI, CII, CIII, CV, CVI, CVII, MI, MII, ER, CP and PX/Po) based on a phylogenetic analysis and subcellular localization. Further structural analysis showed that most of the VvHSP20 genes (93.8%) had no intron or only one intron, while genes that clustered together based on a phylogenetic tree had similar motifs and evolutionarily conserved structures. The HSP20s share a conservedα-crystalline domain (ACD) and the different components of the ACD domain suggest the functional diversity of VvHSP20s. In addition, the 48 VvHSP20 genes were distributed on 12 grape chromosomes and the majority of VvHSP20 genes were located at the proximal or distal ends of chromosomes. Chromosome mapping indicated that four groups of VvHSP20 genes were identified as tandem duplication genes. Phytohormone responsive, abiotic and biotic stress-responsive, and plant development-related cis-elements were identified from the cis-regulatory elements analysis of VvHSP20s. The expression profiles of VvHSP20s genes (VvHSP20-1, 11, 14, 17, 18, 19, 20, 24, 25, 28, 31, 39, 42, and 43) were largely similar between RNA-Seq and qRT-PCR analysis after hydrogen peroxide (H2O2) treatment. The results showed that most VvHSP20s were down-regulated by H2O2 treatment during fruit development. VvHSP20s genes were indeed found to be involved in the grape berry development and differences in their transcriptional levels may be the result of functional differentiation during evolution. CONCLUSIONS: Our results provide valuable information on the evolutionary relationship of genes in the VvHSP20 family, which is useful for future studies on the functional characteristics of VvHSP20 genes in grape.
Assuntos
Genoma de Planta/genética , Proteínas de Choque Térmico HSP20/genética , Peróxido de Hidrogênio/farmacologia , Família Multigênica , Vitis/genética , Frutas/efeitos dos fármacos , Frutas/genética , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Proteínas de Choque Térmico HSP20/efeitos dos fármacos , Filogenia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Estresse Fisiológico , Vitis/efeitos dos fármacos , Vitis/crescimento & desenvolvimentoRESUMO
The current study conducted to investigate the hypothesis that low or excess levels of lipids increased the gill rot morbidity through impairing the immune function and structural integrity in the gill of grass carp (Ctenopharyngodon idella). A total of 540 young grass carp with an average initial weight of 261.41⯱â¯0.53â¯g were fed diets containing six graded levels of lipids at 0.59%, 2.14%, 3.60%, 5.02%, 6.66% and 8.01% diets for 8 weeks. After the growth trial, fish were challenged with Flavobacterium columnare for 3 days. The results indicated that compared with optimal lipids supplementation (2.14%-8.01% lipids diets), low or excess levels of lipids impaired fish immune function through declining the activities of humoral compounds, down-regulated the mRNA levels of anti-inflammatory cytokines, inhibitor of κBα (IκBα) and ribosomal p70S6 kinase (S6K1), and up-regulated pro-inflammatory cytokines, nuclear factor κB p65 (NF-κB p65) (not p52), IκB kinase α (IKKα) (not IKKß), IKKγ and eIF4E-binding protein (4EBP) in the gill of young grass carp. In addition, low or excess levels of lipids decreased young grass carp physical barrier function through down-regulating the mRNA levels of ZO-1 (rather than ZO-2b), Claudin b, c, 3, 12, 15a, 15b, 7b, 7a and Occludin through MAPKK 6/p38 MAPK/MLCK signaling molecules, decreasing antioxidant ability via Kelch-like ECH-associating protein 1a (Keap1a)/NF-E2-related factor 2 (Nrf2) signaling molecules, and down-regulating the mRNA levels of B-cell lymphoma-2 (Bcl-2) and inhibitor of apoptosis protein (IAP) and up-regulating the mRNA levels of apoptotic protease activating factor-1 (Apaf-1), Caspase-3, -8 and -9 and Fas ligand (FasL) in the gill of grass carp. Based on the quadratic regression analysis for the gill rot morbidity, C3 and MDA contents, the dietary lipids requirements for young grass carp have been estimated to be 5.60%, 6.01% and 4.58% diets.
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Ração Animal/análise , Carpas/fisiologia , Gorduras na Dieta , Doenças dos Peixes/imunologia , Infecções por Flavobacteriaceae/veterinária , Brânquias/microbiologia , Animais , Carpas/imunologia , Carpas/microbiologia , Dieta/veterinária , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Infecções por Flavobacteriaceae/imunologia , Infecções por Flavobacteriaceae/microbiologia , Infecções por Flavobacteriaceae/patologia , Flavobacterium/fisiologia , Brânquias/patologia , Imunidade InataRESUMO
Hydrothermal ore deposits at convergent plate boundaries represent extraordinary metal enrichment in the continental crust. They are generally associated with felsic magmatism in extensional settings at high thermal gradients. Although their formation is common during accretionary orogeny, more and more ore deposits have been discovered recently in the collisional orogens of China. Because collisional orogeny was operated in a compressional regime at low thermal gradients, it is not favorable for mobilization of ore-forming elements and thus for the production of hydrothermal ore deposits. Nevertheless, continental collision is generally preceded by oceanic subduction, which enables the preliminary enrichment of ore-forming elements in the mantle wedge due to chemical metasomatism by subducting slab-derived fluids. This gave rise to metal pre-enriched domains in the overriding lithosphere, which may be reactivated by extensional tectonism for hydrothermal mineralization either immediately during accretionary orogeny or at a later time during and after collisional orogeny. It is these tectonic processes that have resulted in the progressive enrichment of ore-forming elements through the geochemical differentiation of the subducting oceanic crust, the metasomatic mantle domains and the mafic juvenile crust, respectively, at different depths. Finally, the reactivation of metal pre-enriched domains by continental rifting in the orogenic lithosphere is the key to the metallogenesis of collisional orogens.
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Van der Waals heterostructures (vdWHs) have opened new avenues for fundamental scientific studies and design of novel devices. Although numerous reports have demonstrated vdWH optoelectronic devices, no report on vdWH lasers can be found to date. In this paper we demonstrated electrically driven vdWH lasers for the first time, and the lasers were realized from ZnO microwire/MgO/p-GaN structures. By coating Ag films on the top surfaces of the ZnO microwires, the current injection and lasing directionality of the vdWH lasers have been improved significantly, and this improvement can be attributed to the high conductivity and reflectivity of the Ag film. The output power of the device can reach 2.41 µW under 14 mA drive current, which is among the highest values ever reported for ZnO based lasers. Our results may provide a promising way to electrically pumped lasers based on micro/nano-structures.
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Adaptation of cyclic brush polymer for drug delivery applications remains largely unexplored. Herein, cyclic brush copolymer of poly(2-hydroxyethyl methacrylate-g-poly(N-isopropylacrylamide-st-N-hydroxyethylacrylamide)) (cb-P(HEMA-g-P(NIPAAm-st-HEAAm))), comprising a cyclic core of PHEMA and thermosensitive brushes of statistical copolymer of P(NIPAAm-st-HEAAm), is designed and synthesized successfully via a graft-from approach using atom transfer free radical polymerization from a cyclic multimacroinitiator. The composition of the brush is optimized to endow the resulting cyclic brush copolymer with a lower critical solution temperature (LCST) slightly above the physiological temperature, but lower than the localized temperature of tumor tissue, which is suitable for the hyperthermia-triggered anticancer drug delivery. Critical aggregation concentration determination reveals better stability for the unimolecular nanoparticle formed by the cyclic brush copolymer than that formed by the bottlebrush analogue. The dramatically increased size with elevated temperatures from below to above the LCST confirms hyperthermia-induced aggregation for both formulations. Such structural destabilization promotes significantly the drug release at 40 °C. Most importantly, the drug-loaded cyclic brush copolymer shows enhanced in vitro cytotoxicity against HeLa cells than the bottlebrush counterpart. The better stability and higher therapeutic efficacy demonstrates that the thermosensitive cyclic brush copolymer is a better formulation than bottle brush copolymer for controlled drug release applications.
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Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos/métodos , Polímeros/química , Temperatura , Resinas Acrílicas/química , Antineoplásicos/química , Antineoplásicos/farmacocinética , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/química , Doxorrubicina/farmacocinética , Portadores de Fármacos/síntese química , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Células HeLa , Humanos , Nanopartículas/química , Nanopartículas/ultraestrutura , Tamanho da Partícula , Poli-Hidroxietil Metacrilato/química , Polímeros/síntese químicaRESUMO
Human noroviruses (HuNoVs) are the dominant cause of food-borne outbreaks of acute gastroenteritis. However, fundamental researches on HuNoVs, such as identification of viral receptors have been limited by the currently immature system to culture HuNoVs and the lack of efficient small animal models. Previously, we demonstrated that the recombinant protruding domain (P domain) of HuNoVs capsid proteins were successfully anchored on the surface of Escherichia coli BL21 cells after the bacteria were transformed with a plasmid expressing HuNoVs P protein fused with bacterial transmembrane anchor protein. The cell-surface-displayed P proteins could specifically recognize and bind to histo-blood group antigens (HBGAs, receptors of HuNoVs). In this study, an upgraded bacterial surface displayed system was developed as a new platform to discover candidate receptors of HuNoVs. A thrombin-susceptible "linker" sequence was added between the sequences of bacterial transmembrane anchor protein and P domain of HuNoV (GII.4) capsid protein in a plasmid that displays the functional P proteins on the surface of bacteria. In this new system, the surface-displayed HuNoV P proteins could be released by thrombin treatment. The released P proteins self-assembled into small particles, which were visualized by electron microscopy. The bacteria with the surface-displayed P proteins were incubated with pig stomach mucin which contained HBGAs. The bacteria-HuNoV P proteins-HBGAs complex could be collected by low speed centrifugation. The HuNoV P proteins-HBGAs complex was then separated from the recombinant bacterial surface by thrombin treatment. The released viral receptor was confirmed by using the monoclonal antibody against type A HBGA. It demonstrated that the new system was able to capture and easily isolate receptors of HuNoVs. This new strategy provides an alternative, easier approach for isolating unknown receptors/ligands of HuNoVs from different samples including mammalian cell lines, oysters, and fresh produce.
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The current study explores the hypothesis that low or excess levels of lipids decrease the enteritis resistance ability through impairing the intestinal physical and immune barrier function of young grass carp (Ctenopharyngodon idella). A total of 540 young grass carp with an average initial weight of 261.41 ± 0.53 g were fed diets containing six graded levels of lipids at 5.9, 21.4, 36.0, 50.2, 66.6 and 80.1 g/kg for 8 weeks. After that, a challenge trial was conducted by injection of Aeromonas hydrophila over 2 weeks. The results show that compared with optimal lipids level, low or excess levels of lipids impair fish immune barrier function through declining humoral compounds and down-regulating the mRNA levels of interleukin 4/13A (IL-4(13)A) [not in the proximal intestine (PI)], IL-4(13)B, IL-6, IL-10, transforming growth factor ß1 (TGF-ß1), inhibitor of κBα (IκBα), TOR and ribosomal p70S6 kinase (S6K1) (P < 0.05), and up-regulating tumor necrosis factor α (TNF-α), interferon γ2 (IFN-γ2), IL-1ß, IL-8, IL-12 p40 (not p35), nuclear factor κB p65 (NF-κB p65), IκB kinase α (IKKα), IKKß, IKKγ, and eIF4E-binding protein (4EBP) in the intestine of young grass carp (P < 0.05). In addition, low or excess levels of lipids also decrease young grass carp physical barrier function through down-regulating the mRNA levels of zonula occludens (ZO-1), ZO-2b [only in the distal intestine (DI)], Claudin b, c, 3, 12, 15a, 15b (only in the DI), 7b (not 7a) and Occludin by MAPKK 6/p38 MAPK (not JNK)/MLCK signaling molecules, down-regulating B-cell lymphoma-2 (Bcl-2) and inhibitor of apoptosis protein (IAP) and up-regulating the mRNA levels of apoptotic protease activating factor-1 (Apaf-1), Caspase-3, -8 and -9 and Fas ligand (FasL) (not in the DI), and decreasing antioxidant ability via Kelch-like ECH-associating protein 1 (Keap1)/NF-E2-related factor 2 (Nrf2) signaling molecules in the intestine. Based on the quadratic regression analysis for the enteritis resistance ability, LA activities and GSH content in the MI were established to be 54.5, 49.91 and 47.83 g lipid/kg diets, respectively.
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Carpas/fisiologia , Doenças dos Peixes/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Imunidade Inata , Metabolismo dos Lipídeos , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Carpas/anatomia & histologia , Carpas/crescimento & desenvolvimento , Carpas/imunologia , Dieta/veterinária , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Intestinos/imunologia , Intestinos/fisiologia , Distribuição AleatóriaRESUMO
Most of the polymers are composed of a crystal part, an amorphous part, and a transitional interfacial part. These components present disparate physical and chemical characteristics. However, it always suffers from peak overlapping in solid NMR spectrum in order to acquire polymer's crystallinity. The polyethylene oxide (PEO) sample was tested using the Torchia pulse sequence combined with the Multivariate Curve Resolution (MCR) method. A two dimensional CP/MAS spectrum containing spin-lattice relaxation time (T1) information was acquired. After the correction based on the reciprocity relation, the overlapped peaks were resolved and quantified together with their T1's. Crystallinity is therefore observed naturally according to components' content ratios associated with their T1 values.
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Our study explored the effect of dietary lipids on growth and immunity and structure (head kidney, spleen and skin) of young grass carp (Ctenopharyngodon idella). A total of 540 young grass carp with an average initial weight of 261.41 ± 0.53 g were fed diets containing six graded levels of lipids at 5.9-80.1 g/kg diet for 8 weeks. After that, a challenge trial was conducted by injection of Aeromonas hydrophila over 2 weeks. The results indicated that compared with optimal lipids supplementation, low and excess levels of lipids down-regulated the mRNA levels of antimicrobial peptides, anti-inflammatory cytokines, inhibitor of κBα (IκBα) and ribosomal p70S6 kinase (S6K1), and up-regulated pro-inflammatory cytokines, nuclear factor κB p65 (NF-κB p65), NF-κB c-Rel (not p52), IκB kinase α (IKKα), IKKß, IKKγ, and eIF4E-binding protein (4EBP) mRNA levels in the head kidney and spleen of young grass carp (P < 0.05). Low or excess levels of lipids also increased reactive oxygen species (ROS) production and malondialdehyde (MDA) and protein carbonyl (PC) contents, reduced the activities of antioxidant enzymes (P < 0.05), down-regulate the relative mRNA levels of antioxidant enzymes and NF-E2-related factor 2 (Nrf2), and up-regulated the expression levels of Kelch-like ECH-associating protein 1a (Keap1a) and Keap1b in the head kidney and spleen. In addition, low or excess levels of lipids down-regulated the mRNA levels of B-cell lymphoma-2 (Bcl-2) and inhibitor of apoptosis protein (IAP) in the head kidney and spleen, whereas up-regulated the mRNA levels of apoptotic protease activating factor-1 (Apaf-1), caspase 3, 7, 8 and 9 mRNA levels in the head kidney and spleen and Fas ligand (FasL) mRNA levels in the spleen of young grass carp, suggesting that low or excess levels of lipids could decrease the head kidney and spleen immune function, induce oxidative damage and apoptosis and impair antioxidant system of young grass carp. At last, low or excess levels of lipids also impaired the immune function and structure in the skin of young grass carp. Based on the quadratic regression analysis for PWG, skin haemorrhage and lesions morbidity and IgM content, the dietary lipids requirements for young grass carp were estimated to be 43.7, 60.2, 55.0 and 52.1 g/kg diet, respectively.
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Carpas/fisiologia , Dieta/veterinária , Doenças dos Peixes/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Imunidade Inata , Metabolismo dos Lipídeos , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Carpas/anatomia & histologia , Carpas/crescimento & desenvolvimento , Carpas/imunologia , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/patologia , Rim Cefálico/microbiologia , Rim Cefálico/patologia , Distribuição Aleatória , Pele/microbiologia , Pele/patologia , Baço/microbiologia , Baço/patologiaRESUMO
Histone lysine methylation is important in early zebrafish development; however, the role of histone arginine methylation in this process remains unclear. H3R2me2a, generated by protein arginine methyltransferase 6 (Prmt6), is a repressive mark. To explore the role of Prmt6 and H3R2me2a during zebrafish embryogenesis, we identified the maternal characteristic of prmt6 and designed two prmt6-specific morpholino-oligos (MOs) to study its importance in early development, application of which led to early epiboly defects and significantly reduced the level of H3R2me2a marks. prmt6 mRNA could rescue the epiboly defects and the H3R2me2a reduction in the prmt6 morphants. Functionally, microarray data demonstrated that growth arrest and DNA damage-inducible, α, a (gadd45αa) was a significantly up-regulated gene in MO-treated embryos, the activity of which was linked to the activation of the p38/JNK pathway and apoptosis. Importantly, gadd45αa MO and p38/JNK inhibitors could partially rescue the defect of prmt6 morphants, the downstream targets of Prmt6, and the apoptosis ratios of the prmt6 morphants. Moreover, the results of ChIP quantitative real time PCR and luciferase reporter assay indicated that gadd45αa is a repressive target of Prmt6. Taken together, these results suggest that maternal Prmt6 is essential to early zebrafish development by directly repressing gadd45αa.
Assuntos
Proteínas de Ciclo Celular/genética , Desenvolvimento Embrionário , Proteínas Nucleares/genética , Proteína-Arginina N-Metiltransferases/genética , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Animais , Apoptose/efeitos dos fármacos , Proteínas de Ciclo Celular/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Morfolinos/farmacologia , Proteínas Nucleares/metabolismo , Proteína-Arginina N-Metiltransferases/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Regulação para Cima/efeitos dos fármacos , Proteínas de Peixe-Zebra/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
OBJECTIVE: To investigate the protective effect of Danxuetong injection (DXT, a combination of Danshen and Xueshuantong injections) against testicular ischemia-reperfusion injury following testis torsion/detorsion in rats. METHODS: Thirty-two 4-week-old healthy male SD rats were randomly divided into four groups of equal number: sham operation, normal saline, single DXT injection, and successive DXT injection. The rat models of testicular ischemia-reperfusion injury were established by 2-hour 720-degree torsion/detorsion of the unilateral testis. At 6 weeks after modeling, the rats were killed and their testes were harvested for measure- ment of testicular coefficients, sperm counts, sperm motility, and the levels of total anti-oxidative capacity (T-AOC) , superoxide dismutase (SOD) , nitric oxide synthase (NOS) , and malondialdehyde ( MDA) in the testis tissue. RESULTS: Compared with the rats of the normal saline group, those of the single DXT injection and successive DXT injection groups showed significant increases in the testicular coefficient (0.11 ± 0.03 vs 0.35 ± 0.04 and 0.40 ± 0.06, P < 0.05), sperm count ([0.46 ± 0.10] vs [1.44 ± 0.50] and [3.00 ± 1.28] x10(9)/ml, P < 0.05), sperm motility ([13.63 ± 14.04] vs [39.63 ± 5.04] and [76.31 ± 3.67]%, P < 0.05), the activity of SOD (72.76 ± 5.58 vs 116.25 ± 8.83 and 133.20 ± 13.84, P < 0.05), and the level of T-AOC (5.58 ± 1.07 vs 13.34 ± 5.81 and 19.21 ± 5.69, P < 0.05), but a remarkable decrease in the content of MDA (42.38 ± 8.94 vs 20.94 ± 5.65 and 15.02 ± 1.03, P < 0. 05) in the injured testes. CONCLUSION: DXT can effectively rid the testis tissue of oxygen free radicals, improve sperm count and motility by antioxidation, and protect the testis tissue of prepubertal rats against testicular ischemia-reperfusion injury after testis torsion/detorsion. It also has a protective effect on the contralateral testis, and successive injection has a better effect than single injection of DXT.