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1.
Genesis ; 62(3): e23603, 2024 06.
Artigo em Inglês | MEDLINE | ID: mdl-38738564

RESUMO

The vomeronasal organ (VNO) is a specialized chemoreceptive structure in many vertebrates that detects chemical stimuli, mostly pheromones, which often elicit innate behaviors such as mating and aggression. Previous studies in rodents have demonstrated that chemical stimuli are actively transported to the VNO via a blood vessel-based pumping mechanism, and this pumping mechanism is necessary for vomeronasal stimulation in behaving animals. However, the molecular mechanisms that regulate the vomeronasal pump remain mostly unknown. In this study, we observed a high level of expression of phosphodiesterase 5A (PDE5A) in the vomeronasal blood vessel of mice. We provided evidence to support the potential role of PDE5A in vomeronasal pump regulation. Local application of PDE5A inhibitors-sildenafil or tadalafil-to the vomeronasal organ (VNO) reduced stimulus delivery into the VNO, decreased the pheromone-induced activity of vomeronasal sensory neurons, and attenuated male-male aggressive behaviors. PDE5A is well known to play a role in regulating blood vessel tone in several organs. Our study advances our understanding of the molecular regulation of the vomeronasal pump.


Assuntos
Nucleotídeo Cíclico Fosfodiesterase do Tipo 5 , Órgão Vomeronasal , Animais , Órgão Vomeronasal/metabolismo , Camundongos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/metabolismo , Nucleotídeo Cíclico Fosfodiesterase do Tipo 5/genética , Masculino , Inibidores da Fosfodiesterase 5/farmacologia , Tadalafila/farmacologia , Citrato de Sildenafila/farmacologia , Feromônios/metabolismo , Agressão/fisiologia , Feminino , Camundongos Endogâmicos C57BL
2.
Plant Cell Rep ; 43(3): 74, 2024 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-38379014

RESUMO

KEY MESSAGE: HanMYB1 was found to play positive roles in the modulation of anthocyanins metabolism based on the integrative analysis of different color cultivars and the related molecular genetic analyses. As a high value ornamental and edible crop with various colors, sunflowers (Helianthus annuus L.) provide an ideal system to understand the formation of flower color. Anthocyanins are major pigments in higher plants, which is associated with development of flower colors and ability of oxidation resistance. Here, we performed an integrative analysis of the transcriptome and flavonoid metabolome in five sunflower cultivars with different flower colors. According to differentially expressed genes and differentially accumulated flavonoids, these cultivars could be grouped into yellow and red. The results showed that more anthocyanins were accumulated in the red group flowers, especially the chrysanthemin. Some anthocyanins biosynthesis-related genes like UFGT (UDP-glycose flavonoid glycosyltransferase) also expressed more in the red group flowers. A MYB transcriptional factor, HanMYB1, was found to play vital positive roles in the modulation of anthocyanins metabolism by the integrative analysis. Overexpressed HanMYB1 in tobacco could deepen the flower color, increase the accumulation of anthocyanins and directly active the express of UFGT genes. Our findings indicated that the MYB transcriptional factors provide new insight into the dynamic regulation of the anthocyanin biosynthesis in facilitating sunflower color formation and anthocyanin accumulation.


Assuntos
Antocianinas , Helianthus , Antocianinas/metabolismo , Transcriptoma/genética , Helianthus/genética , Helianthus/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Flavonoides/metabolismo , Metaboloma , Flores , Regulação da Expressão Gênica de Plantas , Cor , Pigmentação/genética , Perfilação da Expressão Gênica
3.
J Environ Manage ; 356: 120616, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38518493

RESUMO

Metakaolin-based geopolymers are very promising materials for improving the safety of low and intermediate level radioactive waste disposal, with respect to ordinary Portland cement, due to their excellent immobilization performance for Cs+ and superior chemical stability. However, their application is limited by the fact that the leaching behavior of Cs+ is susceptible to the presence of other ions in the environment. Here, we propose a way to modify a geopolymer using perfluorodecyltriethoxysilane (PDFS), successfully reducing the leaching rate of Cs+ in the presence of multiple competitive cations due to blocking the diffusion of water. The leachability index of the modified samples in deionized water and highly concentrated saline water reached 11.0 and 8.0, respectively. The reaction mechanism between PDFS and geopolymers was systematically investigated by characterizing the microstructure and chemical bonding of the material. This work provides a facile and successful approach to improve the immobilization of Cs ions by geopolymers in real complex environments, and it could be extended to further improve the reliability of geopolymers used in a range of applications.


Assuntos
Resíduos Radioativos , Eliminação de Resíduos , Reprodutibilidade dos Testes , Polímeros , Eliminação de Resíduos/métodos , Íons
4.
Plant Biotechnol J ; 21(8): 1671-1681, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37155328

RESUMO

The fungal bioluminescence pathway (FBP) was identified from glowing fungi, which releases self-sustained visible green luminescence. However, weak bioluminescence limits the potential application of the bioluminescence system. Here, we screened and characterized a C3'H1 (4-coumaroyl shikimate/quinate 3'-hydroxylase) gene from Brassica napus, which efficiently converts p-coumaroyl shikimate to caffeic acid and hispidin. Simultaneous expression of BnC3'H1 and NPGA (null-pigment mutant in A. nidulans) produces more caffeic acid and hispidin as the natural precursor of luciferin and significantly intensifies the original fungal bioluminescence pathway (oFBP). Thus, we successfully created enhanced FBP (eFBP) plants emitting 3 × 1011 photons/min/cm2 , sufficient to illuminate its surroundings and visualize words clearly in the dark. The glowing plants provide sustainable and bio-renewable illumination for the naked eyes, and manifest distinct responses to diverse environmental conditions via caffeic acid biosynthesis pathway. Importantly, we revealed that the biosynthesis of caffeic acid and hispidin in eFBP plants derived from the sugar pathway, and the inhibitors of the energy production system significantly reduced the luminescence signal rapidly from eFBP plants, suggesting that the FBP system coupled with the luciferin metabolic flux functions in an energy-driven way. These findings lay the groundwork for genetically creating stronger eFBP plants and developing more powerful biological tools with the FBP system.


Assuntos
Engenharia Metabólica , Plantas , Luciferinas
5.
BMC Cancer ; 23(1): 120, 2023 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-36747161

RESUMO

BACKGROUND: Glioma is characterized by high morbidity, high mortality, and poor prognosis. Despite tremendous advances in the treatment of glioma, the prognosis of patients with glioma is still unsatisfactory. There is an urgent need to discover novel molecular markers that effectively predict prognosis in patients with glioma. The investigation of the role of WEE2-AS1 in various tumors is an emerging research field, but the biological function and prognostic value of WEE2-AS1 in glioma have rarely been reported. This study aimed to assess the value of WEE2-AS1 as a potential prognostic marker of glioma. METHODS: Gene expression (RNA-Seq) data of patients with glioma were extracted from The Cancer Genome Atlas (TCGA) and the Genotype-Tissue Expression (GTEx) databases. The Wilcoxon rank sum test was used to analyze the expression of WEE2-AS1 in the cells and tissues of glioma. The Kruskal-Wallis rank sum test, Wilcoxon rank sum test, and logistic regression were used to evaluate the relationship between clinical variables and expression of WEE2-AS1. Cox regression analysis and the Kaplan-Meier method were used to evaluate the prognostic factors in glioma. A nomogram based on Cox multivariate analysis was used to predict the impact of WEE2-AS1 on glioma prognosis. Gene Set Enrichment Analysis (GSEA) was used to identify key WEE2-AS1-associated signaling pathways. Spearman's rank correlation was used to elucidate the association between WEE2-AS1 expression and immune cell infiltration levels. RESULTS: We found that WEE2-AS1 was overexpressed in a variety of cancers, including glioma. High expression of WEE2-AS1 was associated with glioma progression. We determined that the expression of WEE2-AS1 might be an independent risk factor for the survival and prognosis of patients with glioma. We further observed that the mechanism of WEE2-AS1-mediated tumorigenesis involved neuroactive ligand-receptor interaction, cell cycle, and the infiltration of immune cells into the glioma microenvironment. CONCLUSION: These findings demonstrate that WEE2-AS1 is a promising biomarker for the diagnosis and prognosis of patients with glioma. An increased understanding of its effects on the regulation of cell growth may lead to the development of clinical applications that improve the prognostic status of patients with glioma.


Assuntos
Glioma , RNA Longo não Codificante , Humanos , Carcinogênese , Ciclo Celular , Glioma/genética , Pacientes , Prognóstico , RNA Longo não Codificante/genética , Microambiente Tumoral/genética
6.
Genomics ; 114(2): 110309, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35131477

RESUMO

Daphnia sinensis is a widespread freshwater microcrustacean. The assembled D. sinensis genome totaled 131.58 Mb with 92.23% of the assembly anchored onto 10 chromosomes. Based on the whole genome information, we further compared the transcriptomic and epigenomic characterization among parthenogenetic females, sexual females and males in D. sinensis. Transcriptomic analysis showed that the up-regulated genes in males were mainly grouped into the cuticle, sex differentiation and methyl farnesoate synthesis, which might play a pivotal role in steering development and reproduction processes. By comparison, the highly expressed genes in parthenogenetic females were mainly grouped into energy metabolism, mitosis, and DNA replication, which might contribute to maintaining rapid production of parthenogenetic females, and nutrient uptake for the growth of neonates. The whole-genome DNA methylation analysis showed that the methylation rate in parthenogenetic females was higher than that in sexual females and males, which might contribute to its rapid response to environment stress.


Assuntos
Daphnia , Reprodução , Animais , Biologia , Daphnia/genética , Feminino , Masculino , Partenogênese/genética , Reprodução/genética , Diferenciação Sexual
7.
J Environ Manage ; 345: 118836, 2023 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-37634403

RESUMO

Soil salinization can affect the ecological environment of soil and alter greenhouse gas (GHG) emissions. Chitooligosaccharides and Arbuscular mycorrhizal fungi (AMF) reduced the GHG fluxes of salinized soil, and this reduction was attributed to an alteration in the rhizosphere microecology, including changes in the activities of ß-glucosidase, acid phosphatase, N-acetyl-ß-D-glucosidase, and Leucine aminopeptidase. Additionally, certain bacteria species such as paracoccus, ensifer, microvirga, and paracyclodium were highly correlated with GHG emissions. Another interesting finding is that foliar spraying of chitooligosaccharides could transport to the soybean root system, and improve soybean tolerance to salt stress. This is achieved by enhancing the activities of antioxidant enzymes, and the changes in amino acid metabolism, lipid metabolism, and membrane transport. Importantly, the Co-application of chitooligosaccharides and Arbuscular mycorrhiza fungi was found to have a greater effect compared to their application alone.


Assuntos
Gases de Efeito Estufa , Micorrizas , Glycine max , Rizosfera , Solo/química , Raízes de Plantas , Fungos , Microbiologia do Solo
8.
Molecules ; 28(15)2023 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-37570807

RESUMO

Myelin repair, which is known as remyelination, is critical to the treatment of neurodegenerative diseases, and myelination depends on not only the differentiation of oligodendrocyte precursor cells toward oligodendrocytes but also the renewal of oligodendrocyte precursor cells under pathological conditions. However, simultaneously promoting the differentiation and proliferation of oligodendrocyte precursor cells in lesions remains an unmet challenge and might affect demyelinating diseases. Kidney-tonifying herbs of traditional Chinese medicine (TCM) are effective in improving the symptoms of degenerative patients. However, herbs or compounds with dual functions are unverified. The purpose of this study was to find a kidney-tonifying TCM that synchronously improved the differentiation and proliferation of oligodendrocyte precursor cells under pathological conditions. Compounds with dual functions were screened from highly frequently used kidney-tonifying TCM, and the effects of the obtained compound on remyelination were investigated in an in vitro oligodendrocyte precursor cell differentiation model under pathological conditions and in demyelinating mice in vivo. The compound icaritin, which is an active component of Yin-Yang-Huo (the leaves of Epimedium brevicornu Maxim), demonstrated multiple effects on the remyelination process, including enhancing oligodendrocyte precursor cell proliferation, facilitating the differentiation of neural progenitor cells toward oligodendrocyte precursor cells and further toward oligodendrocytes, and maturation of oligodendrocytes under corticosterone- or glutamate-induced pathological conditions. Importantly, icaritin effectively rescued behavioral functions and increased the formation of myelin in a cuprizone-induced demyelination mouse model. The multiple effects of icaritin make it a promising lead compound for remyelination therapy.


Assuntos
Doenças Desmielinizantes , Células Precursoras de Oligodendrócitos , Camundongos , Animais , Células Precursoras de Oligodendrócitos/patologia , Doenças Desmielinizantes/induzido quimicamente , Doenças Desmielinizantes/tratamento farmacológico , Doenças Desmielinizantes/patologia , Diferenciação Celular , Proliferação de Células , Camundongos Endogâmicos C57BL
9.
J Am Chem Soc ; 144(15): 6802-6812, 2022 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-35378978

RESUMO

Density functional theory calculations (ωB97X-D) are reported for the reactions of methoxy, tert-butoxy, trichloroethoxy, and trifluoroethoxy radicals with a series of 26 C-H bonds in different environments characteristic of a variety of hydrocarbons and substituted derivatives. The variations in activation barriers are analyzed with modified Evans-Polanyi treatments to account for polarity and unsaturation effects. The treatments by Roberts and Steel and by Mayer have inspired the development of a simple treatment involving the thermodynamics of reactions, the difference between the reactant radical and product radical electronegativities, and the absence or presence of α-unsaturation. The three-parameter equation (ΔH⧧ = 0.52ΔHrxn(1 - d) - 0.35ΔχAB2 + 10.0, where d = 0.44 when there is α-unsaturation to the reacting C-H bond), correlates well with quantum mechanically computed barriers and shows the quantitative importance of the thermodynamics of reactions (dictated by the reactant and the product bond dissociation energies) and polar effects.


Assuntos
Hidrocarbonetos , Hidrogênio , Radicais Livres/química , Hidrocarbonetos/química , Hidrogênio/química , Termodinâmica
10.
Plant Biotechnol J ; 20(9): 1743-1755, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35587579

RESUMO

Water is crucial for plant growth and survival. The transcellular water movement is facilitated by aquaporins (AQPs) that rapidly and reversibly modify water permeability. The abundance of AQPs is regulated by its synthesis, redistribution and degradation. However, the molecular mechanism of proteasomal degradation of AQPs remains unclear. Here, we demonstrate that a novel E3 ligase, OsRINGzf1, mediated the degradation of AQPs in rice. OsRINGzf1 is the candidate gene from a drought-related quantitative trait locus (QTL) on the long arm of chromosome 4 in rice (Oryza sativa) and encodes a Really Interesting New Gene (RING) zinc finger protein 1. OsRINGzf1 possesses the E3 ligase activity, ubiquitinates and mediates OsPIP2;1 degradation, thus reducing its protein abundance. The content of OsPIP2;1 protein was decreased in OsRINGzf1 overexpression (OE) plants. The degradation of OsPIP2;1 was inhibited by MG132. The OsRINGzf1 OE plants, with higher leaf-related water content (LRWC) and lower leaf water loss rate (LWLR), exhibited enhanced drought resistance, whereas the RNAi and knockout plants of OsRINGzf1 were more sensitive to drought. Together, our data demonstrate that OsRINGzf1 positively regulates drought resistance through promoting the degradation of OsPIP2;1 to enhance water retention capacity in rice.


Assuntos
Aquaporinas , Oryza , Aquaporinas/genética , Aquaporinas/metabolismo , Secas , Regulação da Expressão Gênica de Plantas/genética , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética , Ubiquitina/genética , Ubiquitina/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Água/metabolismo
11.
Nanomedicine ; 45: 102591, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35907618

RESUMO

The efficacy of Adoptive Cell Therapy (ACT) for solid tumor is still mediocre. This is mainly because tumor cells can hijack ACT T cells' immune checkpoint pathways to exert immunosuppression in the tumor microenvironment. Immune Checkpoint Inhibitors such as anti-PD-1 (aPD1) can counter the immunosuppression, but the synergizing effects of aPD1 to ACT was still not satisfactory. Here we demonstrate an approach to safely anchor aPD1-formed nanogels onto T cell surface via bio-orthogonal click chemistry before adoptive transfer. The spatial-temporal co-existence of aPD1 with ACT T cells and the responsive drug release significantly improved the treatment outcome of ACT in murine solid tumor model. The average tumor weight of the group treated by cell-surface anchored aPD1 was only 18 % of the group treated by equivalent dose of free aPD1 and T cells. The technology can be broadly applicable in ACTs employing natural or Chimeric Antigen Receptor (CAR) T cells.


Assuntos
Neoplasias , Receptores de Antígenos Quiméricos , Animais , Terapia Baseada em Transplante de Células e Tecidos , Inibidores de Checkpoint Imunológico , Imunoterapia Adotiva , Camundongos , Nanogéis , Neoplasias/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Microambiente Tumoral
12.
Ecotoxicol Environ Saf ; 247: 114273, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36356529

RESUMO

Sterigmatocystin (STE) is a common hepatotoxic and nephrotoxic contaminant in cereals, however, its phytotoxicity and mechanisms are poorly understood. Here, the phytotoxic mechanisms of STE were investigated via the metabolomics of Amaranthus retroflexus L. A total of 140 and 113 differential metabolites were detected in the leaves and stems, respectively, among which amino acids, lipids, and phenolic compounds were significantly perturbed. Valine, leucine, isoleucine, and lysine biosynthesis were affected by STE. These metabolic responses revealed that STE might be toxic to plants by altering the plasma membrane and inducing oxidative damage, which was verified by measuring the relative electrical conductivity and quantification of reactive oxygen species. The elevated amino acids, as well as the decreased of D-sedoheptuiose-7-phosphate indicated increased proteolysis and carbohydrate metabolism restriction. Furthermore, the IAA level also decreased. This study provides a better understanding of the impacts of STE on the public health, environment and food security.


Assuntos
Alcaloides , Amaranthus , Toxinas Biológicas , Esterigmatocistina , Metabolômica , Aminoácidos
13.
IEEE Trans Circuits Syst Video Technol ; 32(10): 6642-6656, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37215187

RESUMO

Video captioning is a challenging task as it needs to accurately transform visual understanding into natural language description. To date, state-of-the-art methods inadequately model global-local vision representation for sentence generation, leaving plenty of room for improvement. In this work, we approach the video captioning task from a new perspective and propose a GLR framework, namely a global-local representation granularity. Our GLR demonstrates three advantages over the prior efforts. First, we propose a simple solution, which exploits extensive vision representations from different video ranges to improve linguistic expression. Second, we devise a novel global-local encoder, which encodes different video representations including long-range, short-range and local-keyframe, to produce rich semantic vocabulary for obtaining a descriptive granularity of video contents across frames. Finally, we introduce the progressive training strategy which can effectively organize feature learning to incur optimal captioning behavior. Evaluated on the MSR-VTT and MSVD dataset, we outperform recent state-of-the-art methods including a well-tuned SA-LSTM baseline by a significant margin, with shorter training schedules. Because of its simplicity and efficacy, we hope that our GLR could serve as a strong baseline for many video understanding tasks besides video captioning. Code will be available.

14.
Atmos Environ (1994) ; 264: 118713, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34522157

RESUMO

In this work, we use observations and experimental emissions in a version of NOAA's National Air Quality Forecasting Capability to show that the COVID-19 economic slowdown led to disproportionate impacts on near-surface ozone concentrations across the contiguous U.S. (CONUS). The data-fusion methodology used here includes both U.S. EPA Air Quality System ground and the NASA Aura satellite Ozone Monitoring Instrument (OMI) NO2 observations to infer the representative emissions changes due to the COVID-19 economic slowdown in the U.S. Results show that there were widespread decreases in anthropogenic (e.g., NOx) emissions in the U.S. during March-June 2020, which led to widespread decreases in ozone concentrations in the rural regions that are NOx-limited, but also some localized increases near urban centers that are VOC-limited. Later in June-September, there were smaller decreases, and potentially some relative increases in NOx emissions for many areas of the U.S. (e.g., south-southeast) that led to more extensive increases in ozone concentrations that are partly in agreement with observations. The widespread NOx emissions changes also alters the O3 photochemical formation regimes, most notably the NOx emissions decreases in March-April, which can enhance (mitigate) the NOx-limited (VOC-limited) regimes in different regions of CONUS. The average of all AirNow hourly O3 changes for 2020-2019 range from about +1 to -4 ppb during March-September, and are associated with predominantly urban monitoring sites that demonstrate considerable spatiotemporal variability for the 2020 ozone changes compared to the previous five years individually (2015-2019). The simulated maximum values of the average O3 changes for March-September range from about +8 to -4 ppb (or +40 to -10%). Results of this work have implications for the use of widespread controls of anthropogenic emissions, particularly those from mobile sources, used to curb ozone pollution under the current meteorological and climate conditions in the U.S.

15.
Molecules ; 26(6)2021 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-33809536

RESUMO

Pyrrolizidine alkaloids (PAs) are a class of natural toxins with hepatotoxicity, genotoxicity and carcinogenicity. They are endogenous and adulterated toxic components widely found in food and herbal products. In this study, a sensitive and efficient ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was used to detect the PAs in 386 kinds of Chinese herbal medicines recorded in the Chinese Pharmacopoeia (2020). The estimated daily intake (EDI) of 0.007 µg/kg body weight (bw)/day was adopted as the safety baseline. The margin of exposure (MOE) approach was applied to evaluate the chronic exposure risk for the genotoxic and carcinogenic potential of PAs. Results showed that PAs was detected in 271 out of 386 samples with a content of 0.1-25,567.4 µg/kg, and there were 20 samples with EDI values above the baseline, 0.007 µg/kg bw/day. Beyond that, the MOE values for 10 out of 271 positive samples were below 10,000. Considering the actual situation, Haber's rule was used to assume two weeks exposure every year during lifetime, and still the MOE values for four out of 271 positive samples were under 10,000, indicating these products may have potential health risk. The developed method was successfully applied to detect the PAs-containing Chinese herbal medicines. This study provides convincing data that can support risk management actions in China and a meaningful reference for the rational and safe use of Chinese herbal medicines.


Assuntos
Medicamentos de Ervas Chinesas/química , Alcaloides de Pirrolizidina/química , Carcinógenos/química , China , Cromatografia Líquida de Alta Pressão/métodos , Medicina Herbária/métodos , Humanos , Medição de Risco , Espectrometria de Massas em Tandem/métodos
16.
J Neuroophthalmol ; 40(1): 22-29, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31246675

RESUMO

PURPOSE: Clinical trials of gene therapy for Leber hereditary optic neuropathy (LHON) were conducted in 9 volunteers with the mitochondrial mutation, G11778A in ND4. The purpose of this study was to investigate whether multilocus mitochondrial mutations directly influence the efficacy of gene therapy for LHON. METHODS: Nine volunteers with LHON participated in a clinical trial with intravitreal injection of an adenoviral vector expressing wild-type ND4. Patients were subsequently divided into 2 groups: according to the differences in therapy efficacy and based on improvements in visual acuity. Full mitochondrial DNA sequences of the 2 groups of patients were generated and compared using PubMed, PolyPhen, and PROVEAN. Furthermore, the association between the detected mutations and clinical effects of gene therapy was analyzed. RESULTS: Best-corrected visual acuity (BCVA) significantly improved (≥0.3 log of minimum angle of resolution [logMAR]) in 7 patients 6 months after gene therapy, whereas there was no significant change in BCVA (<0.3 logMAR) of the remaining 2 patients. All 9 patients carried the G1178A mutation in addition to other nonsynonymous mutations. Among these mutations, some were predicted to be neutral and deleterious. Meanwhile, different mitochondrial mutations in the group in which treatment was ineffective, compared with those in responders, were at nucleotide positions 6569 (CO1; Patient 3), 9641 (CO3; Patient 3), and 4491 (ND2; Patient 5). CONCLUSIONS: Detection of the 3 primary mitochondrial mutations causing LHON is sufficient for screening before gene therapy; sequencing of the entire mitochondrial genome is unnecessary before treatment. Patients with LHON can respond to targeted gene therapy irrespective of additional multilocus mitochondrial mutations.


Assuntos
DNA Mitocondrial/genética , Terapia Genética/métodos , Mitocôndrias/genética , Mutação , Atrofia Óptica Hereditária de Leber/terapia , Acuidade Visual/fisiologia , Adolescente , Adulto , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Atrofia Óptica Hereditária de Leber/genética , Atrofia Óptica Hereditária de Leber/fisiopatologia , Resultado do Tratamento , Adulto Jovem
17.
Plant Cell ; 28(9): 2161-2177, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27468891

RESUMO

Plants have evolved complicated protective mechanisms to survive adverse conditions. Previously, we reported that the transcription factor OsbZIP46 regulates abscisic acid (ABA) signaling-mediated drought tolerance in rice (Oryza sativa) by modulating stress-related genes. An intrinsic D domain represses OsbZIP46 activity, but the detailed mechanism for the repression of OsbZIP46 activation remains unknown. Here, we report an OsbZIP46-interacting protein, MODD (Mediator of OsbZIP46 deactivation and degradation), which is homologous to the Arabidopsis thaliana ABSCISIC ACID-INSENSITIVE5 binding protein AFP. MODD was induced by ABA and drought stress, but the induction was much slower than that of OsbZIP46 In contrast to OsbZIP46, MODD negatively regulates ABA signaling and drought tolerance, and inhibits the expression of OsbZIP46 target genes. We found that MODD negatively regulates OsbZIP46 activity and stability. MODD represses OsbZIP46 activity via interaction with the OsTPR3-HDA702 corepressor complex and downregulation of the histone acetylation level at OsbZIP46 target genes. MODD promotes OsbZIP46 degradation via interaction with the U-box type ubiquitin E3 ligase OsPUB70. Interestingly, the D domain is required for both deactivation and degradation of OsbZIP46 via its interaction with MODD. These findings show that plants fine-tune their drought responses by elaborate regulatory mechanisms, including the coordination of activity and stability of key transcription factors.

18.
Plant Biotechnol J ; 16(8): 1415-1423, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29327438

RESUMO

Recently, CRISPR-Cas (clustered, regularly interspaced short palindromic repeats-CRISPR-associated proteins) system has been used to produce plants resistant to DNA virus infections. However, there is no RNA virus control method in plants that uses CRISPR-Cas system to target the viral genome directly. Here, we reprogrammed the CRISPR-Cas9 system from Francisella novicida to confer molecular immunity against RNA viruses in Nicotiana benthamiana and Arabidopsis plants. Plants expressing FnCas9 and sgRNA specific for the cucumber mosaic virus (CMV) or tobacco mosaic virus (TMV) exhibited significantly attenuated virus infection symptoms and reduced viral RNA accumulation. Furthermore, in the transgenic virus-targeting plants, the resistance was inheritable and the progenies showed significantly less virus accumulation. These data reveal that the CRISPR/Cas9 system can be used to produce plant that stable resistant to RNA viruses, thereby broadening the use of such technology for virus control in agricultural field.


Assuntos
Francisella/genética , Doenças das Plantas/virologia , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/virologia , Vírus de RNA/patogenicidade , Sistemas CRISPR-Cas , Doenças das Plantas/imunologia , Plantas Geneticamente Modificadas/metabolismo , Vírus de RNA/imunologia
19.
Plant Physiol ; 173(2): 1475-1491, 2017 02.
Artigo em Inglês | MEDLINE | ID: mdl-28062835

RESUMO

Cold stress is one of the major limiting factors for rice (Oryza sativa) productivity. Several MYB transcriptional factors have been reported as important regulators in the cold stress response, but the molecular mechanisms are largely unknown. In this study, we characterized a cold-responsive R2R3-type MYB gene, OsMYB30, for its regulatory function in cold tolerance in rice. Functional analysis revealed that overexpression of OsMYB30 in rice resulted in increased cold sensitivity, while the osmyb30 knockout mutant showed increased cold tolerance. Microarray and quantitative real-time polymerase chain reaction analyses revealed that a few ß-amylase (BMY) genes were down-regulated by OsMYB30. The BMY activity and maltose content, which were decreased and increased in the OsMYB30 overexpression and osmyb30 knockout mutant, respectively, were correlated with the expression patterns of the BMY genes. OsMYB30 was shown to bind to the promoters of the BMY genes. These results suggested that OsMYB30 exhibited a regulatory effect on the breakdown of starch through the regulation of the BMY genes. In addition, application of maltose had a protective effect for cell membranes under cold stress conditions. Furthermore, we identified an OsMYB30-interacting protein, OsJAZ9, that had a significant effect in suppressing the transcriptional activation of OsMYB30 and in the repression of BMY genes mediated by OsMYB30. These results together suggested that OsMYB30 might be a novel regulator of cold tolerance through the negative regulation of the BMY genes by interacting with OsJAZ9 to fine-tune the starch breakdown and the content of maltose, which might contribute to the cold tolerance as a compatible solute.


Assuntos
Oryza/fisiologia , Proteínas de Plantas/metabolismo , Estresse Fisiológico , beta-Amilase/metabolismo , Membrana Celular/metabolismo , Temperatura Baixa , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Maltose/metabolismo , Mutação , Oryza/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Amido/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , beta-Amilase/genética
20.
Ophthalmic Res ; 60(3): 176-184, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29649796

RESUMO

PURPOSE: To assess changes in visual field (VF) values after gene therapy for Leber's hereditary optic neuropathy (LHON). METHODS: VF recovery, VF indices, and mean deviation in injected and uninjected eyes, before and after gene therapy, were examined in 2 groups of patients according to disease duration (≤2 years and > 2 years). Nine patients with LHON were treated by monocular intravitreal injection of AAV2-ND4. Finally, 7 patients were considered for subsequent comparisons; the first and second eyes were treated separately. RESULTS: There were no significant differences in VF indices and mean deviation between injected and uninjected eyes (p = 0.910 and p = 0.929, respectively). However, there was a significant difference before and after injection (p = 0.016 and p = 0.015, respectively). There was no significant difference in VF improvement between patients with ≤2 years' disease duration and those with a longer disease duration. CONCLUSION: There was a statistically significant VF improvement after gene therapy. This suggests that monocular intravitreal injection of AAV2-ND4 can improve binocular VF values. This study also suggests that gene therapy can be effective in patients with a disease duration of > 2 years.


Assuntos
Terapia Genética , Atrofia Óptica Hereditária de Leber/terapia , Campos Visuais/fisiologia , Adolescente , Adulto , Análise de Variância , Criança , Feminino , Vetores Genéticos/administração & dosagem , Humanos , Injeções Intravítreas , Masculino , Pessoa de Meia-Idade , Atrofia Óptica Hereditária de Leber/fisiopatologia , Acuidade Visual/fisiologia , Adulto Jovem
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