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1.
Artigo em Inglês | MEDLINE | ID: mdl-38698748

RESUMO

AIMS: This study investigates the impact of IbACP (Ipomoea batatas anti-cancer peptide) on defense-related gene expression in tomato leaves, focusing on its role in plant defense mechanisms. BACKGROUND: Previously, IbACP was isolated from sweet potato leaves, and it was identified as a peptide capable of inducing an alkalinization response in tomato suspension culture media. Additionally, IbACP was found to regulate the proliferation of human pancreatic adenocarcinoma cells. OBJECTIVE: Elucidate IbACP's molecular influence on defense-related gene expression in tomato leaves using next-generation sequencing analysis. METHOD: To assess the impact of IbACP on defense-related gene expression, transcriptome data were analyzed, encompassing various functional categories such as photosynthesis, metabolic processes, and plant defense. Semi-quantitative reverse-transcription polymerase chain reaction analysis was employed to verify transcription levels of defense-related genes in tomato leaves treated with IbACP for durations ranging from 0 h (control) to 24 h. RESULTS: IbACP induced jasmonic acid-related genes (LoxD and AOS) at 2 h, with a significant up-regulation of salicylic acid-dependent gene NPR1 at 24 h. This suggested a temporal antagonistic effect between jasmonic acid and salicylic acid during the early hours of IbACP treatment. Downstream ethylene-responsive regulator genes (ACO1, ETR4, and ERF1) were consistently down-regulated by IbACP at all times. Additionally, IbACP significantly up-regulated the gene expressions of suberization-associated anionic peroxidases (TMP1 and TAP2) at all time points, indicating enhanced suberization of the plant cell wall to prevent pathogen invasion. CONCLUSION: IbACP enhances the synthesis of defense hormones and up-regulates downstream defense genes, improving the plant's resistance to biotic stresses.

2.
Nanomedicine (Lond) ; 18(16): 1045-1059, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37610004

RESUMO

Background: The use of nanodiamonds (NDs) and fluorescent nanodiamonds (FNDs) as nonallergenic biocompatible additives in incomplete Freund's adjuvant (IFA) to elicit immune responses in vivo was investigated. Methods: C57BL/6 mice were immunized with chicken egg ovalbumin (OVA) in IFA and also OVA-conjugated NDs (or OVA-conjugated FNDs) in IFA to produce antibodies. OVA-expressing E.G7 lymphoma cells and OVA-negative EL4 cells were inoculated in mice to induce tumor formation. Results: The new formulation significantly enhanced immune responses and thus disease resistance. It exhibited specific therapeutic activities, effectively inhibiting the growth of E.G7 tumor cells in mice over 35 days. Conclusion: The high biocompatibility and multiple functionalities of NDs/FNDs render them applicable as active and trackable vaccine adjuvants and antitumor agents.


Assuntos
Nanodiamantes , Animais , Camundongos , Camundongos Endogâmicos C57BL , Emulsões , Ovalbumina , Corantes , Vacinação
3.
Biol Res ; 56(1): 7, 2023 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-36843032

RESUMO

BACKGROUND: The distinct arterial and venous cell fates are dictated by a combination of various genetic factors which form diverse types of blood vessels such as arteries, veins, and capillaries. We report here that YULINK protein is involved in vasculogenesis, especially venous formation. METHODS: In this manuscript, we employed gene knockdown, yeast two-hybrid, FLIM-FRET, immunoprecipitation, and various imaging technologies to investigate the role of YULINK gene in zebrafish and human umbilical vein endothelial cells (HUVECs). RESULTS: Knockdown of YULINK during the arterial-venous developmental stage of zebrafish embryos led to the defective venous formation and abnormal vascular plexus formation. Knockdown of YULINK in HUVECs impaired their ability to undergo cell migration and differentiation into a capillary-like tube formation. In addition, the phosphorylated EPHB4 was decreased in YULINK knockdown HUVECs. Yeast two-hybrid, FLIM-FRET, immunoprecipitation, as well as imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B or TICAM2) and markers (Clathrin and RHOB). VEGF-induced VEGFR2 internalization was also compromised in YULINK knockdown HUVECs, demonstrating to the involvement of YULINK. CONCLUSION: This study suggests that YULINK regulates vasculogenesis, possibly through endocytosis in zebrafish and HUVECs.


Assuntos
Saccharomyces cerevisiae , Peixe-Zebra , Animais , Humanos , Células Endoteliais da Veia Umbilical Humana , Peixe-Zebra/genética , Movimento Celular , Diferenciação Celular , Neovascularização Fisiológica
4.
Biol. Res ; 56: 7-7, 2023. ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1429908

RESUMO

BACKGROUND: The distinct arterial and venous cell fates are dictated by a combination of various genetic factors which form diverse types of blood vessels such as arteries, veins, and capillaries. We report here that YULINK protein is involved in vasculogenesis, especially venous formation. METHODS: In this manuscript, we employed gene knockdown, yeast two-hybrid, FLIM-FRET, immunoprecipitation, and various imaging technologies to investigate the role of YULINK gene in zebrafish and human umbilical vein endothelial cells (HUVECs). RESULTS: Knockdown of YULINK during the arterial-venous developmental stage of zebrafish embryos led to the defective venous formation and abnormal vascular plexus formation. Knockdown of YULINK in HUVECs impaired their ability to undergo cell migration and differentiation into a capillary-like tube formation. In addition, the phosphorylated EPHB4 was decreased in YULINK knockdown HUVECs. Yeast two-hybrid, FLIM-FRET, immunoprecipitation, as well as imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B or TICAM2) and markers (Clathrin and RHOB). VEGF-induced VEGFR2 internalization was also compromised in YULINK knockdown HUVECs, demonstrating to the involvement of YULINK. CONCLUSION: This study suggests that YULINK regulates vasculogenesis, possibly through endocytosis in zebrafish and HUVECs. Key points Knockdown of YULINK with morpholino in embryos of double transgenic zebrafish exhibited abnormal venous formation. Tube formation and phosphorylated EPHB4 were decreased in YULINK knockdown HUVECs. FLIM-FRET, immunoprecipitation, as well as other imaging technologies showed that YULINK colocalized with endosome related proteins (EPS15, RAB33B and TICAM2) and endosome markers (Clathrin and RHOB). Knockdown of YULINK decreased the internalization of VEGF and VEGFR2 in HUVECs.


Assuntos
Humanos , Animais , Saccharomyces cerevisiae , Peixe-Zebra/genética , Diferenciação Celular , Movimento Celular , Neovascularização Fisiológica , Células Endoteliais da Veia Umbilical Humana
5.
Sci Rep ; 12(1): 21139, 2022 12 07.
Artigo em Inglês | MEDLINE | ID: mdl-36477410

RESUMO

Coriander (Coriandrum sativum L.) contains abundant antioxidants and essential oils which can provide antibacterial, antifungal, and antioxidant activities in the pharmaceutical, health and food production industry. To improve the economic values of coriander, the relationships between optimal light treatments for maximizing both plant growth and the antioxidant and essential oil content of coriander leaves need to be determined. Plants were exposed to five light-emitting diodes spectral color mixtures, high blue light (BL) intensity induced the levels of reducing power response. The light treatments were then adjusted for the analysis of secondary metabolite compounds of coriander leaves. Among 30 identified compounds, the amounts of decamethyl-cyclopentasiloxane and dodecane were significantly reduced in the R80 + G50 + B50 condition, whereas dodecamethyl-cyclohexasiloxane level was significantly reduced in R50 + G50 + B80 condition. Various light quality and intensity combinations influenced the accumulations of chlorophyll and phytochemical contents, mediated antioxidative properties, and secondary metabolites of coriander leaves, which may be useful in developing a new LED lighting apparatus optimized for coriander production in plant factories.


Assuntos
Coriandrum , Antioxidantes
6.
Plants (Basel) ; 11(15)2022 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-35956541

RESUMO

Sweet potato (Ipomoea batatas L.) is one of the most important food crops worldwide, with leaves of different varieties showing purple, green and yellow, and these leaves provide a dietary source of nutrients and various bioactive compounds. The objective of this study was to identify the active constituents of chlorogenic acids (CGAs) in different methanolic extract of leaves of three varieties of sweet potato (purple CYY 98-59, green Taoyuan 2, and yellow CN 1927-16) using liquid chromatography-tandem mass spectrometry. Genotype-specific metabolite variations were observed; CGAs and three isomeric peaks were detected in sweet potato leaf extracts (SPLEs). Among them, the yellow SPLE contained the highest contents of 3,5-dicaffeoylquinic acid (3,5-di-CQA) and 3,4-dicaffeoylquinic acid (3,4-di-CQA), followed by the green SPLE, whereas the purple SPLE retained lower 3,5-di-CQA content compared to yellow and green SPLEs. All three SPLEs contained lower 4,5-dicaffeoylquinic acid (4,5-di-CQA) and CGA contents compared to 3,5-di-CQA and 3,4-di-CQA, although CGA constituents were not significantly different in genotypes, whereas purple SPLE contained higher 4,5-di-CQA content compared to yellow and green SPLEs. This study indicates that SPLs marketed in Taiwan vary widely in their biological potentials and may impart different health benefits to consumers.

7.
Opt Lett ; 47(15): 3676-3679, 2022 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-35913287

RESUMO

Integrated photonics provides a path for miniaturization of an optical system to a compact chip scale and offers reconfigurability by the integration of active components. Here we report a chip-scale reconfigurable scan lens based on an optical phased array, consisting of 30 actively controlled elements on the InP integrated photonic platform. By configuring the phase shifters, we show scanning of a nearly diffraction-limited focused spot with a full width at half maximum spot size down to 2.7 µm at the wavelength of 1550 nm. We demonstrate the key functions needed for a laser-scanning microscope, including light focusing, collection, and steering. We also perform confocal measurements to detect reflection at selective depths.

8.
Bioeng Transl Med ; 7(2): e10285, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35600645

RESUMO

Monocytes are important regulators for the maintenance of homeostasis in innate and adaptive immune system and have been reported to play important role in cancer progression. CD47-SIRPα recognition is a coinhibitory immune signal to inhibit phagocytosis in monocytes and macrophages and has been well-known as the "Don't eat me" signal. By using an approach of integrated sensing and activating proteins (iSNAPs), we have rewired the CD47-SIRPα axis to create iSNAP-M which activates pathways in engineered human monocytes (iSNAP-MC). The mRNA expression levels of the monocyte/macrophage markers CD11b, CD14, and CD31 are upregulated in iSNAP-monocytes (iSNAP-MC). With PMA induction, the iSNAP-MC-derived macrophages (iSNAP-MΦ) showed upregelation in CD86 and CD80, but not CD206. TNFα expression and secretion were also increased in iSNAP-MΦ. Furthermore, the injection of iSNAP-MC into mice bearing human B-lymphoma tumors led to the suppression of tumor progression. Therefore, the engineered monocytes, via blockage of coinhibitory immune signals by rewiring CD47-SIRPα axis, can be applied to suppress target tumors for cancer immunotherapy.

9.
Int J Biol Macromol ; 211: 316-327, 2022 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-35568153

RESUMO

Increasing trends in food safety awareness drive consumer demands for fresher healthier diets and has led to the development of low-cost pH-sensitive indicator films to evaluate deterioration levels in fresh foods. Arrowroot starch/iota-carrageenan-based films were combined with 10, 30, 50% Kyoho skin extract (KSE) to produce indicator films with halochromic abilities. The KSE indicator films were characterized based on their physico-mechanical, functional, and crystallinity properties; thermal stability; and their pH-dependent color changes in in situ anthocyanin-based monitoring of shrimp freshness. All KSE indicator films displayed compact structure under scanning electron microscope analysis and increased tensile strength, exhibited UV-vis barrier ability, and presented low water wettability. Moreover, FTIR signaled strong hydrogen bond interactions among polymers and KSE that strengthened peak crystallinity in XRD analysis and lowered weight loss at melting temperature, which indicated thermal stability of the indicator films. Furthermore, pH-sensitivity of the indicator films integrated with natural KSE anthocyanin demonstrated color changes from purple to red under acidic conditions, purple to green in an ammonium environment, and yellow appeared in high alkaline conditions. Finally, this study demonstrated the food packaging and halochromic capacity of biopolymer-based pH-sensitivity of the KSE indicator films in real-time monitoring of shrimp at room storage temperatures.


Assuntos
Marantaceae , Antocianinas/química , Carragenina/química , Embalagem de Alimentos , Concentração de Íons de Hidrogênio , Extratos Vegetais/química , Amido/química
10.
PLoS One ; 17(4): e0265311, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35363791

RESUMO

Deer genera around the globe are threatened by anthropogenic interference. The translocation of alien species and their subsequent genetic introgression into indigenous deer populations is particularly harmful to the species of greatest conservation concern. Products derived from deer, including venison and antler velvet, are also at risk of fraudulent labeling. The current molecular markers used to genetically identify deer species were developed from genome sequences and have limited applicability for cross-species amplification. The absence of efficacious diagnostic techniques for identifying deer species has hampered conservation and wildlife crime investigation efforts. Expressed sequence tag-simple sequence repeat (EST-SSR) markers are reliable tools for individual and species identification, especially in terms of cross-species genotyping. We conducted transcriptome sequencing of sambar (Rusa unicolor) antler velvet and acquired 11,190 EST-SSRs from 65,074 newly assembled unigenes. We identified a total of 55 unambiguous amplicons in sambar (n = 45), which were selected as markers to evaluate cross-species genotyping in sika deer (Cervus nippon, n = 30) and red deer (Cervus elaphus, n = 46), resulting in cross-species amplification rates of 94.5% and 89.1%, respectively. Based on polymorphic information content (>0.25) and genotyping fidelity, we selected 16 of these EST-SSRs for species identification. This marker set revealed significant genetic differentiation based on the fixation index and genetic distance values. Principal coordinate analysis and STRUCTURE analysis revealed distinct clusters of species and clearly identified red-sika hybrids. These markers showed applicability across different genera and proved suitable for identification and phylogenetic analyses across deer species.


Assuntos
Chifres de Veado , Cervos , Animais , Cervos/genética , Etiquetas de Sequências Expressas , Repetições de Microssatélites/genética , Filogenia
11.
Plant Physiol ; 188(1): 442-459, 2022 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-34747472

RESUMO

Chloroplasts are the sites for photosynthesis, and two Golden2-like factors act as transcriptional activators of chloroplast development in rice (Oryza sativa L.) and maize (Zea mays L.). Rice OsGLK1 and OsGLK2 are orthologous to maize ZmGLK1 (ZmG1) and ZmGLK2 (ZmG2), respectively. However, while rice OsGLK1 and OsGLK2 act redundantly to regulate chloroplast development in mesophyll cells, maize ZmG1 and ZmG2 are functionally specialized and expressed in different cell-specific manners. To boost rice chloroplast development and photosynthesis, we generated transgenic rice plants overexpressing ZmG1 and ZmG2, individually or simultaneously, with constitutive promoters (pZmUbi::ZmG1 and p35S::ZmG2) or maize promoters (pZmG1::ZmG1, pZmG2::ZmG2, and pZmG1::ZmG1/pZmG2::ZmG2). Both ZmG1 and ZmG2 genes were highly expressed in transgenic rice leaves. Moreover, ZmG1 and ZmG2 showed coordinated expression in pZmG1::ZmG1/pZmG2::ZmG2 plants. All Golden2-like (GLK) transgenic plants had higher chlorophyll and protein contents, Rubisco activities and photosynthetic rates per unit leaf area in flag leaves. However, the highest grain yields occurred when maize promoters were used; pZmG1::ZmG1, pZmG2::ZmG2, and pZmG1::ZmG1/pZmG2::ZmG2 transgenic plants showed increases in grain yield by 51%, 47%, and 70%, respectively. In contrast, the pZmUbi::ZmG1 plant produced smaller seeds without yield increases. Transcriptome analysis indicated that maize GLKs act as master regulators promoting the expression of both photosynthesis-related and stress-responsive regulatory genes in both rice shoot and root. Thus, by promoting these important functions under the control of their own promoters, maize GLK1 and GLK2 genes together dramatically improved rice photosynthetic performance and productivity. A similar approach can potentially improve the productivity of many other crops.


Assuntos
Cloroplastos/genética , Cloroplastos/metabolismo , Oryza/crescimento & desenvolvimento , Oryza/genética , Fotossíntese/genética , Sementes/crescimento & desenvolvimento , Sementes/genética , Zea mays/genética , Produtos Agrícolas/genética , Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Genótipo , Fatores de Transcrição/genética
13.
J Inorg Biochem ; 225: 111602, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34547604

RESUMO

In this focused review, we portray the recently reported 2.5 Å cyro-EM structure of the particulate methane monooxygenase (pMMO) from M. capsulatus (Bath). The structure of the functional holo-pMMO near atomic resolution has uncovered the sites of the copper cofactors including the location of the active site in the enzyme. The three coppers seen in the original X-ray crystal structures of the enzyme are now augmented by additional coppers in the transmembrane domain as well as in the water-exposed C-terminal subdomain of the PmoB subunit. The cryo-EM structure offers the first glimpse of the catalytic machinery capable of methane oxidation with high selectivity and efficiency. The findings are entirely consistent with the biochemical and biophysical findings previously reported in the literature, including the chemistry of hydrocarbon hydroxylation, regeneration of the catalyst for multiple turnovers, and the mechanism of aborting non-productive cycles to ensure kinetic competence.


Assuntos
Metano/química , Oxigenases/química , Biocatálise , Domínio Catalítico , Cobre/química , Hidroquinonas/química , Methylococcus capsulatus/enzimologia , NAD/química , Oxirredução , Conformação Proteica em alfa-Hélice , Domínios Proteicos , Subunidades Proteicas/química , Ubiquinona/análogos & derivados , Ubiquinona/química
14.
BMC Biol ; 19(1): 214, 2021 09 24.
Artigo em Inglês | MEDLINE | ID: mdl-34560855

RESUMO

BACKGROUND: Yeast one-hybrid (Y1H) is a common technique for identifying DNA-protein interactions, and robotic platforms have been developed for high-throughput analyses to unravel the gene regulatory networks in many organisms. Use of these high-throughput techniques has led to the generation of increasingly large datasets, and several software packages have been developed to analyze such data. We previously established the currently most efficient Y1H system, meiosis-directed Y1H; however, the available software tools were not designed for processing the additional parameters suggested by meiosis-directed Y1H to avoid false positives and required programming skills for operation. RESULTS: We developed a new tool named GateMultiplex with high computing performance using C++. GateMultiplex incorporated a graphical user interface (GUI), which allows the operation without any programming skills. Flexible parameter options were designed for multiple experimental purposes to enable the application of GateMultiplex even beyond Y1H platforms. We further demonstrated the data analysis from other three fields using GateMultiplex, the identification of lead compounds in preclinical cancer drug discovery, the crop line selection in precision agriculture, and the ocean pollution detection from deep-sea fishery. CONCLUSIONS: The user-friendly GUI, fast C++ computing speed, flexible parameter setting, and applicability of GateMultiplex facilitate the feasibility of large-scale data analysis in life science fields.


Assuntos
Saccharomyces cerevisiae , Análise de Dados , Redes Reguladoras de Genes , Robótica , Saccharomyces cerevisiae/genética , Software
15.
Nanomaterials (Basel) ; 11(6)2021 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-34204514

RESUMO

Nanodiamond (ND) has recently emerged as a potential nanomaterial for nanovaccine development. Here, a plant-based haemagglutinin protein (H5.c2) of A/H5N1 virus was conjugated with detonation NDs (DND) of 3.7 nm in diameter (ND4), and high-pressure and high-temperature (HPHT) oxidative NDs of ~40-70 nm (ND40) and ~100-250 nm (ND100) in diameter. Our results revealed that the surface charge, but not the size of NDs, is crucial to the protein conjugation, as well as the in vitro and in vivo behaviors of H5.c2:ND conjugates. Positively charged ND4 does not effectively form stable conjugates with H5.c2, and has no impact on the immunogenicity of the protein both in vitro and in vivo. In contrast, the negatively oxidized NDs (ND40 and ND100) are excellent protein antigen carriers. When compared to free H5.c2, H5.c2:ND40, and H5.c2:ND100 conjugates are highly immunogenic with hemagglutination titers that are both 16 times higher than that of the free H5.c2 protein. Notably, H5.c2:ND40 and H5.c2:ND100 conjugates induce over 3-folds stronger production of both H5.c2-specific-IgG and neutralizing antibodies against A/H5N1 than free H5.c2 in mice. These findings support the innovative strategy of using negatively oxidized ND particles as novel antigen carriers for vaccine development, while also highlighting the importance of particle characterization before use.

16.
Anal Chem ; 93(18): 7140-7147, 2021 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-33913330

RESUMO

The negatively charged nitrogen-vacancy center in fluorescent nanodiamonds (FNDs) is a point defect with unique magneto-optical properties. It emits far-red fluorescence at ∼700 nm, and its intensity can be magnetically modulated with a depth of more than 10% at a field strength of 30 mT. We have closely examined this property and illustrated its practical use in biomedicine by applying a periodic, time-varying magnetic field to FNDs deposited on a surface or dispersed in a solution with a lock-in detection method. We achieved selective and sensitive detection of 100 nm FNDs on a nitrocellulose membrane at a particle density of 0.04 ng/mm2 (or ∼2 × 104 particles/mm2) and in an aqueous solution with a particle concentration of 1 ng/mL (or ∼1 fM) in 10 s as the detection limits. The utility and versatility of the technique were demonstrated with an application to background-free detection of FNDs as reporters for FND-based lateral flow immunoassays as well as selective quantification of FNDs in tissue digests for in vivo studies.


Assuntos
Nanodiamantes , Fluorescência , Nitrogênio
17.
Molecules ; 26(7)2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-33804903

RESUMO

Sweet potato (Ipomoea batatas) is one of the most important food crops worldwide and its leaves provide a dietary source of nutrients and various bioactive compounds. These constituents of sweet potato leaves (SPL) vary among varieties and play important roles in treating and preventing various diseases. Recently, more attentions in health-promoting benefits have led to several in vitro and in vivo investigations, as well as the identification and quantification of bioactive compounds in SPL. Among them, many new compounds have been reported as the first identified compounds from SPL with their dominant bioactivities. This review summarizes the current knowledge of the bioactive compositions of SPL and their health benefits. Since SPL serve as a potential source of micronutrients and functional compounds, they can be further developed as a sustainable crop for food and medicinal industries.


Assuntos
Antioxidantes/química , Produtos Agrícolas/química , Ipomoea batatas/química , Compostos Fitoquímicos/química , Folhas de Planta/química , Antioxidantes/uso terapêutico , Compostos Fitoquímicos/uso terapêutico
18.
Plant Sci ; 305: 110849, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33691955

RESUMO

IbACP, Ipomoea batatas anti-cancer peptide, a sixteen-amino-acid peptide isolated from sweet potato leaves, is capable of mediating a rapid alkalinization of growth media in plant suspension cells. However, the biological roles of IbACP as a defense peptide have not been studied. The objective of this study was to investigate the effect of IbACP on the accumulation of reactive oxygen species (ROS) and the expression of the defense-related genes. IbACP treatment of sweet potato leaves resulted in marked accumulation of both superoxide ion (O2-) and hydrogen peroxide (H2O2). The activity of peroxidase (POD) was significantly enhanced by IbACP treatment, suggesting that high levels of POD antioxidant activity may be used to scavenge the excess H2O2 in sweet potato plants. The IbACP-related genes were identified by suppression subtractive hybridization (SSH), and were then classified and assigned to the following categories: defense, development, metabolism, signaling, gene expression, and abiotic stress. H2O2 acts as a second messenger for gene activation in some of the IbACP-triggered gene expressions. These results demonstrated that IbACP is part of an integrated strategy for genetic regulation in sweet potato. Our work highlights the function of IbACP and its potential use for enhancing stress tolerance in sweet potato, in an effort to improve our understanding of defense-response mechanisms.


Assuntos
Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Peptídeos/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Estresse Fisiológico/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Variação Genética , Genótipo , Ipomoea batatas/química , Folhas de Planta/química , Proteínas de Plantas/genética
19.
Front Bioinform ; 1: 788308, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-36303748

RESUMO

The functions of biological macromolecules are often associated with conformational malleability of the structures. This phenomenon of chemically identical molecules with different structures is coined structural polymorphism. Conventionally, structural polymorphism is observed directly by structural determination at the density map level from X-ray crystal diffraction. Although crystallography approach can report the conformation of a macromolecule with the position of each atom accurately defined in it, the exploration of structural polymorphism and interpreting biological function in terms of crystal structures is largely constrained by the crystal packing. An alternative approach to studying the macromolecule of interest in solution is thus desirable. With the advancement of instrumentation and computational methods for image analysis and reconstruction, cryo-electron microscope (cryo-EM) has been transformed to be able to produce "in solution" structures of macromolecules routinely with resolutions comparable to crystallography but without the need of crystals. Since the sample preparation of single-particle cryo-EM allows for all forms co-existing in solution to be simultaneously frozen, the image data contain rich information as to structural polymorphism. The ensemble of structure information can be subsequently disentangled through three-dimensional (3D) classification analyses. In this review, we highlight important examples of protein structural polymorphism in relation to allostery, subunit cooperativity and function plasticity recently revealed by cryo-EM analyses, and review recent developments in 3D classification algorithms including neural network/deep learning approaches that would enable cryo-EM analyese in this regard. Finally, we brief the frontier of cryo-EM structure determination of RNA molecules where resolving the structural polymorphism is at dawn.

20.
J Microbiol Immunol Infect ; 54(4): 616-626, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32127288

RESUMO

BACKGROUND: Salmonella enterica serovar Typhimurium, a non-typhoidal food-borne pathogen, causes acute enterocolitis, bacteremia, extraintestinal focal infections in humans. Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) contribute to invading into host cellular cytosol, residing in Salmonella-containing vacuoles for intracellular survival, and inducing cellular apoptosis. This study aimed to better understand the mechanism underlying apoptosis in Salmonella-infected macrophages. METHODS: S. Typhimurium SL1344 was used to evaluate extrinsic and intrinsic apoptosis pathways in THP-1 monocyte-derived macrophages in response to Salmonella infection. RESULTS: Activated caspase-3-induced apoptosis pathways, including extrinsic (caspase-8-mediated) and intrinsic (caspase-9-mediated) pathways, in Salmonella-infected macrophages were verified. THP-1 cells with dysfunction of TLR-4 and TLR-5 and Salmonella SPI-1 and SPI-2 mutants were constructed to identify the roles of the genes associated with programmed cell death in the macrophages. Caspase-3 activation in THP-1 macrophages was induced by Salmonella through TLR-4 and TLR-5 signaling pathways. We also identified that SPI-1 structure protein PrgH and effectors SipB and SipD, but not SPI-2 structure protein SsaV, could induce apoptosis via caspase-3 activation and reduce the secretion of inflammation marker TNF-α in the Salmonella-infected cells. The two effectors also reduced the translocation of the p65 subunit of NF-κB into the nucleus and the expression of TNF-α, and then inflammation was diminished. CONCLUSION: Non-typhoid Salmonella induced apoptosis of macrophages and thereby reduced inflammatory cytokine production through the expression of SPI-1. This mechanism in host-pathogen interaction may explain why Salmonella usually manifests as occult bacteremia with less systemic inflammatory response syndrome in the bloodstream infection of children.


Assuntos
Apoptose , Ilhas Genômicas/genética , Interações Hospedeiro-Patógeno , Macrófagos/microbiologia , Macrófagos/patologia , Salmonella typhimurium/genética , Caspase 3/genética , Caspase 3/imunologia , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Humanos , Inflamação , Macrófagos/imunologia , Salmonella typhimurium/imunologia , Salmonella typhimurium/patogenicidade , Células THP-1 , Virulência
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