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Advances in omics technologies now permit the generation of highly contiguous genome assemblies, detection of transcripts and metabolites at the level of single cells and high-resolution determination of gene regulatory features. Here, using a complementary, multi-omics approach, we interrogated the monoterpene indole alkaloid (MIA) biosynthetic pathway in Catharanthus roseus, a source of leading anticancer drugs. We identified clusters of genes involved in MIA biosynthesis on the eight C. roseus chromosomes and extensive gene duplication of MIA pathway genes. Clustering was not limited to the linear genome, and through chromatin interaction data, MIA pathway genes were present within the same topologically associated domain, permitting the identification of a secologanin transporter. Single-cell RNA-sequencing revealed sequential cell-type-specific partitioning of the leaf MIA biosynthetic pathway that, when coupled with a single-cell metabolomics approach, permitted the identification of a reductase that yields the bis-indole alkaloid anhydrovinblastine. We also revealed cell-type-specific expression in the root MIA pathway.
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Antineoplásicos , Catharanthus , Plantas Medicinais , Catharanthus/genética , Plantas Medicinais/metabolismo , Multiômica , Alcaloides Indólicos/metabolismo , Antineoplásicos/metabolismo , Monoterpenos/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Plants produce an extraordinary array of natural products (specialized metabolites). Notably, these structurally complex molecules are not evenly distributed throughout plant tissues but are instead synthesized and stored in specific cell types. Elucidating both the biosynthesis and function of natural products would be greatly facilitated by tracking the location of these metabolites at the cell-level resolution. However, detection, identification, and quantification of metabolites in single cells, particularly from plants, have remained challenging. Here, we show that we can definitively identify and quantify the concentrations of 16 molecules from four classes of natural products in individual cells of leaf, root, and petal of the medicinal plant Catharanthus roseus using a plate-based single-cell mass spectrometry method. We show that identical natural products show substantially different patterns of cell-type localization in different tissues. Moreover, we show that natural products are often found in a wide range of concentrations across a population of cells, with some natural products at concentrations of over 100 mM per cell. This single-cell mass spectrometry method provides a highly resolved picture of plant natural product biosynthesis partitioning at a cell-specific resolution.
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Healing of large calvarial bone defects remains challenging. An RNA-guided Split dCas12a system is previously harnessed to activate long non-coding RNA H19 (lncRNA H19, referred to as H19 thereafter) in bone marrow-derived mesenchymal stem cells (BMSCs). H19 activation in BMSCs induces chondrogenic differentiation, switches bone healing pathways, and improves calvarial bone repair. Since adipose-derived stem cells (ASCs) can be harvested more easily in large quantity, here it is aimed to use ASCs as an alternative cell source. However, H19 activation alone using the Split dCas12a system in ASCs failed to elicit evident chondrogenesis. Therefore, split dCas12a activators are designed more to co-activate other chondroinductive transcription factors (Sox5, Sox6, and Sox9) to synergistically potentiate differentiation. It is found that co-activation of H19/Sox5/Sox6 in ASCs elicited more potent chondrogenic differentiation than activation of Sox5/Sox6/Sox9 or H19 alone. Co-activating H19/Sox5/Sox6 in ASCs significantly augmented in vitro cartilage formation and in vivo calvarial bone healing. These data altogether implicated the potentials of the Split dCas12a system to trigger multiplexed gene activation in ASCs for differentiation pathway reprogramming and tissue regeneration.
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Diferenciação Celular , Condrogênese , RNA Longo não Codificante , Fatores de Transcrição SOXD , Crânio , Fatores de Transcrição SOXD/metabolismo , Fatores de Transcrição SOXD/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Animais , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Tecido Adiposo/citologia , Células-Tronco/metabolismo , Células-Tronco/citologia , Osteogênese/genéticaRESUMO
BACKGROUND: Ovarian stimulation and the use of human chorionic gonadotropin (hCG) for triggering oocyte maturation in women undergoing in vitro fertilisation (IVF) introduces several differences in luteal phase hormone levels compared with natural cycles that may negatively impact on endometrial receptivity and pregnancy rates after fresh embryo transfer. Exogenous luteal phase support is given to overcome these issues. The suitability of a pragmatic approach to luteal phase support is not known due to a lack of data on early phase luteal hormone levels and their association with fertility outcomes during IVF with fresh embryo transfer. This study determined early luteal phase profiles of serum progesterone, 17-hydroxyprogesterone and hCG, and associations between hormone levels/hormone level profile after hCG trigger and the live birth rate in women undergoing IVF with fresh embryo transfer. METHODS: This prospective single center, cohort study was conducted in Vietnam from January 2021 to December 2022. Women aged 18-38 years with normal ovarian reserve and undergoing controlled ovarian stimulation using a gonadotropin-releasing hormone antagonist protocol were included. Serum hormone levels were determined before trigger, at 12, 24 and 36 h after hCG, and daily from 1 to 6 days after oocyte pick-up. Serum hormone level profiles were classified as lower or upper. The primary outcome was live birth rate based on early luteal phase hormone level profile. RESULTS: Ninety-five women were enrolled. Live birth occurred in 19/69 women (27.5%) with a lower progesterone profile and 13/22 (59.1%) with an upper progesterone profile (risk ratio [RR] 2.15; 95% confidence interval [CI] 1.28-3.60), and in 6/31 (19.4%) versus 26/60 (43.3%) with a lower versus upper serum 17-hydroxyprogesterone profile (RR 2.24; 95% CI 1.03-4.86). Nearly 20% of women had peak progesterone concentration on or before day 3 after oocyte pick-up, and this was associated with significantly lower chances of having a life birth. CONCLUSIONS: These data show the importance of proper corpus luteum function with sufficient progesterone/17-hydroxyprogesterone production for achievement of pregnancy and to maximize the chance of live birth during IVF. TRIAL REGISTRATION: NCT04693624 ( www. CLINICALTRIALS: gov ).
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Gonadotropina Coriônica , Fertilização in vitro , Fase Luteal , Indução da Ovulação , Progesterona , Humanos , Feminino , Fase Luteal/sangue , Fase Luteal/fisiologia , Fertilização in vitro/métodos , Adulto , Gravidez , Estudos Prospectivos , Progesterona/sangue , Gonadotropina Coriônica/administração & dosagem , Indução da Ovulação/métodos , Taxa de Gravidez , Adulto Jovem , 17-alfa-Hidroxiprogesterona/sangue , Estudos de Coortes , Transferência Embrionária/métodos , Adolescente , Coeficiente de Natalidade , Resultado do Tratamento , Nascido Vivo/epidemiologiaRESUMO
BACKGROUND: The optimal treatment for malignant gastric outlet obstruction (GOO) remains uncertain. This systematic review aimed to comprehensively investigate the efficacy and safety of four palliative treatments for malignant GOO: gastrojejunostomy, endoscopic ultrasound-guided gastroenterostomy (EUS-GE), stomach-partitioning gastrojejunostomy (PGJ), and endoscopic stenting. METHODS: We searched PubMed, Embase, Cochrane Library, Scopus, and Web of Science databases, ClinicalTrials.gov, and the World Health Organization International Clinical Trials Registry Platform for randomized controlled trials (RCTs) and cohort studies comparing the four treatments for malignant GOO. We included studies that reported at least one of the following clinical outcomes: clinical success, 30-day mortality, reintervention rate, or length of hospital stay. Evidence from RCTs and non-RCTs was naïve combined to perform network meta-analysis through the frequentist approach using an inverse variance model. Treatments were ranked by P score. RESULTS: This network meta-analysis included 3617 patients from 4 RCTs, 4 prospective cohort studies, and 32 retrospective cohort studies. PGJ was the optimal approach in terms of clinical success and reintervention (P scores: 0.95 and 0.90, respectively). EUS-GE had the highest probability of being the optimal treatment in terms of 30-day mortality and complications (P scores: 0.82 and 0.99, respectively). Cluster ranking to combine the P scores for 30-day mortality and reintervention indicated the benefits of PGJ and EUS-GE (cophenetic correlation coefficient: 0.94; PGJ and EUS-GE were in the same cluster). CONCLUSION: PGJ and EUS-GE are recommended for malignant GOO. PGJ could be the alternative choice in centers with limited resources or in patients who are unsuitable for EUS-GE.
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RESEARCH QUESTION: What is the diagnostic accuracy of hysterosalpingo-foam sonography (HyFoSy), using two-dimensional ultrasound in tubal patency assessment in infertile women compared with laparoscopy with dye chromotubation? DESIGN: This prospective study was conducted at My Duc Hospital, Vietnam. Infertile women aged 18 years or older, who were scheduled for laparoscopy, were included. Visual Analogue Scale (VAS) score for perception of pain during HyFoSy was used. Laparoscopy was carried out on the same day. Clinicians undertaking laparoscopy were blinded to HyFoSy results. Sensitivity, specificity, negative and positive predictive value, and 95% confidence intervals were calculated. A sample size of 455 women (nâ¯=â¯910 fallopian tubes) was needed to demonstrate a fluctuation hypothesis, not exceeding 6%, for sensitivity and specificity (power 0.80, two-sided alpha 5%, loss to follow-up 5%). RESULTS: Between 2019 and 2022, 455 participants were recruited. Hysterosalpingo-foam sonography was unsuccessfully carried out in six participants. Two withdrew their consent. Data analysis was conducted on the remaining 447 participants (nâ¯=â¯868 fallopian tubes). The sensitivity and specificity of hysterosalpingo-foam sonography compared with laparoscopy were 0.75 (95% CI 0.71 to 0.79) and 0.70 (95% CI 0.65 to 0.74), respectively. Hysterosalpingo-foam sonography gave a positive predictive value of 0.76 (95% CI 0.73 to 0.80) and negative predictive value of 0.68 (95% CI 0.64 to 0.73). A total of 42.8% of women reported a VAS score of no pain. No adverse event was reported. CONCLUSION: Compared with laparoscopy with dye chromotubation, two-dimensional HyFoSy is a well-tolerated, reliable technique for assessing tubal patency.
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Testes de Obstrução das Tubas Uterinas , Tubas Uterinas , Infertilidade Feminina , Laparoscopia , Ultrassonografia , Humanos , Feminino , Infertilidade Feminina/diagnóstico por imagem , Adulto , Tubas Uterinas/diagnóstico por imagem , Ultrassonografia/métodos , Testes de Obstrução das Tubas Uterinas/métodos , Estudos Prospectivos , Sensibilidade e Especificidade , Adulto Jovem , Histerossalpingografia/métodosRESUMO
In this study, magnetic material based on graphene oxide (GO) was developed for enhanced adsorption capacity for heavy metals. The Fe3O4nanoparticles were combined with the GO material using a chitosan (CS) binder to obtain the CS-Fe3O4/GO nanocomposite. The adsorption capacity of this nanocomposite was evaluated by removing heavy metals including Ni2+ions. When GO was composed with Fe3O4and CS, the GO films were densely covered with ferromagnetic particles, which were bound and densely distributed on the GO film surface due to the interaction between GO and CS. The optimal conditions for the complexation of Ni2+and 4-(2-pyridyl azo)-rezoxine (PAR) are 1 ml Ni2+, 2 ml PAR 100 mg l-1,pH= 6 (adjusted with 0.7 ml of the 0.1 M K2HPO4solution) and a complexation time of 20 min. After 50 min of adsorption, the Ni2+removal efficiency of the CS-Fe3O4/GO nanocomposite reached 81.21% and the corresponding adsorption capacity was 2.03 mg g-1. The Ni2+removal process followed the first-order model and Freundlich isotherm. This process was spontaneous (ΔGo< 0) and an exothermic process (ΔHo= -1128.875 J·mol-1). In addition, the factors affecting this process were investigated, including thepHsolution, the dosage of the CS-Fe3O4/GO nanocomposite and the initial Ni2+concentration. The CS-Fe3O4/GO nanocomposite showed a potential adsorption capacity in removing Ni2+at low concentrations from wastewater.
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Normal-phase (NP) liquid chromatography is one of the most effective methods for separating isomers with sensitive structural features, including xanthophyll isomers. In this work, reverse-phase (RP) and NP liquid chromatography (LC), with silica gel and diol phase, respectively, were evaluated for the separation of xanthophyll isomers. The results showed that RP LC with monomeric C18 phase not only poorly separate all xanthophyll isomers in egg yolk but also requires additional sample preparation to eliminate triacylglycerols in egg yolk. The diol phase of NP-LC provided the highest efficiency for separating lutein, zeaxanthin, and their cis-isomers with isocratic separation using mobile phases consisting of n-hexane and polar modifiers (such as acetone, methyl tert-butyl ether, or ethyl acetate). To determine the xanthophyll content, peak areas from LC and total absorbance from spectrophotometry measurements were used. The approach was applied to analyze the xanthophylls of nine commercial egg samples. The results revealed that five out of nine analyzed samples contained a high level of canthaxanthin, which contributes to color enhancement but not to prevent age-related macular degeneration. Together, it shows that NP LC with diol phase combined with spectrophotometry is a powerful tool to monitor xanthophylls in eggs.
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Gema de Ovo , Xantofilas , Gema de Ovo/química , Cromatografia Líquida de Alta Pressão , Xantofilas/análise , Xantofilas/química , Espectrofotometria , Animais , IsomerismoRESUMO
The primary transcript structure provides critical insights into protein diversity, transcriptional modification, and functions. Cassava transcript structures are highly diverse because of alternative splicing (AS) events and high heterozygosity. To precisely determine and characterize transcript structures, fully sequencing cloned transcripts is the most reliable method. However, cassava annotations were mainly determined according to fragmentation-based sequencing analyses (e.g., EST and short-read RNA-seq). In this study, we sequenced the cassava full-length cDNA library, which included rare transcripts. We obtained 8,628 non-redundant fully sequenced transcripts and detected 615 unannotated AS events and 421 unannotated loci. The different protein sequences resulting from the unannotated AS events tended to have diverse functional domains, implying that unannotated AS contributes to the truncation of functional domains. The unannotated loci tended to be derived from orphan genes, implying that the loci may be associated with cassava-specific traits. Unexpectedly, individual cassava transcripts were more likely to have multiple AS events than Arabidopsis transcripts, suggestive of the regulated interactions between cassava splicing-related complexes. We also observed that the unannotated loci and/or AS events were commonly in regions with abundant single nucleotide variations, insertions-deletions, and heterozygous sequences. These findings reflect the utility of completely sequenced FLcDNA clones for overcoming cassava-specific annotation-related problems to elucidate transcript structures. Our work provides researchers with transcript structural details that are useful for annotating highly diverse and unique transcripts and alternative splicing events.
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Processamento Alternativo , Manihot , Processamento Alternativo/genética , Manihot/genética , Manihot/metabolismo , Nucleotídeos , Biblioteca Gênica , Sequência de BasesRESUMO
Both ZnO and TiO2are common semiconducting metal oxides with high mechanical and chemical durability. However, they only have good photocatalytic ability in the UV region, besides the rapid recombination between electrons and holes reduces the efficiency of the decomposition of organic substances. To improve their catalytic efficiency, in this study, ZnO and TiO2were doped with B to produce the novel B/ZnO/TiO2nanocomposites for degrading tetracycline hydrochloride (TCH) in the aqueous solution. The characteristics of samples were analyzed by the diffuse reflectance ultraviolet-visible (DR/UV-vis), scanning electron microscope (SEM), energy-dispersive (EDS), Fourier transform infrared spectroscopy (FT-IR), and x-ray diffraction (XRD) techniques. The 3B/ZnO/TiO2sample had a band gap energy (Eg) of 3.21 eV. Although the B/ZnO/TiO2sample had a tightly aggregated morphology composed of many nanoparticles in 33-137 nm, it still exhibited a higher uniformly and photocatalytic efficiency than ZnO and ZnO/TiO2. At the optimal doped B of 3 wt%, the degradation efficiency (DE) was achieved at 96.33% with a rate constant of 0.067 min-1. The factors that affect the photocatalytic process such as the initial TCH concentration, the catalyst content, and the pH solution were comprehensively investigated. In addition, the stability of 3B/ZnO/TiO2nanocomposite was evaluated via three consecutive cycles and the DE was 69.75% in 3rd cycle. The Z-scheme mechanism was proposed for the photocatalytic mechanism of TCH in the B/ZnO/TiO2catalyst. In addition, electrical energy consumption was estimated that the electrical energy per order only was 29.05 kW.h.l-1.
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Carnation-like ZnO was synthesized by the facile precipitation method (at room temperature and in 120 min) to decompose dyes in an aqueous medium. The carnation-like ZnO had a stratified porous structure with a size of about 2-3µm, its petals had a smooth surface with a thickness of 5-10 nm and a width of about 300-500 nm. Ag-ZnO composites were synthesized using glucose with the assistance of PVP. The morphology of Ag-ZnO composites was almost unchanged compared to ZnO. Where, the Ag nanoparticles in the size range of 5-15 nm were uniformly dispersed on the ZnO petals, improving the catalytic ability of the composites in tartrazine (TA) degradation. The influence of Ag content on catalytic structure and performance of composite was studied. The 5Ag-ZnO sample had the highest BET surface area and pore volume and the lowest gap energy (Eg) among the as-synthesized samples. The 5Ag-ZnO sample proclaimed the degradation efficiency in 70 min of 97.8% and thekapof 0.031 min-1. The influences of catalyst content, solution pH, and concentration of dye on the photodegradation efficiency of the composite were thoroughly studied. Besides, the photocatalytic activity of the composite was demonstrated by degrading various organic substances and reusability. In addition, it was compared to a metal-semiconductor catalyst of Au-ZnO and semiconductor-semiconductor catalysts of MoS2-ZnO, Cu2O-ZnO, and SiO2-ZnO. The catalytic mechanism under visible light was proposed.
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Calvarial bone healing is challenging, especially for individuals with osteoporosis because stem cells from osteoporotic patients are highly prone to adipogenic differentiation. Based on previous findings that chondrogenic induction of adipose-derived stem cells (ASCs) can augment calvarial bone healing, we hypothesized that activating chondroinductive Sox Trio genes (Sox5, Sox6, Sox9) and repressing adipoinductive genes (C/ebp-α, Ppar-γ) in osteoporotic ASCs can reprogram cell differentiation and improve calvarial bone healing after implantation. However, simultaneous gene activation and repression in ASCs is difficult. To tackle this problem, we built a CRISPR-BiD system for bi-directional gene regulation. Specifically, we built a CRISPR-AceTran system that exploited both histone acetylation and transcription activation for synergistic Sox Trio activation. We also developed a CRISPR interference (CRISPRi) system that exploited DNA methylation for repression of adipoinductive genes. We combined CRISPR-AceTran and CRISPRi to form the CRISPR-BiD system, which harnessed three mechanisms (transcription activation, histone acetylation, and DNA methylation). After delivery into osteoporotic rat ASCs, CRISPR-BiD significantly enhanced chondrogenesis and in vitro cartilage formation. Implantation of the engineered osteoporotic ASCs into critical-sized calvarial bone defects significantly improved bone healing in osteoporotic rats. These results implicated the potential of the CRISPR-BiD system for bi-directional regulation of cell fate and regenerative medicine.
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Regeneração Óssea , Condrogênese , Tecido Adiposo , Animais , Regeneração Óssea/genética , Diferenciação Celular/genética , Condrogênese/genética , Humanos , Ratos , Células-Tronco , Ativação TranscricionalRESUMO
BACKGROUNDS: The COVID-19 pandemic has resulted in not only significant mortalities in Vietnam but has had an impact on its economy. Previous studies have highlighted how the pandemic has had a marginal impact on Vietnamese healthcare workers working at the frontlines. To date, there have been several other studies examining the impact of COVID-19 on intentions to transition between jobs among healthcare professionals, but this has yet to be explored amongst Vietnamese healthcare workers. METHODS: To achieve the study's objectives an online cross-sectional study was conducted between September to November 2021. Snowball sampling methodology was adopted for the recruitment of participants. The questionnaire that was used for this study comprised of the following sections: (a) socio-demographic information; (b) impact of COVID-19 on work; (c) risk of exposure to COVID-19; (d) career choices/intentions to change job, and (e) motivation at work. RESULTS: There were 5727 completed the entire survey. 17.2% of the respondents have had increased job satisfaction, 26.4% reported increased motivation to work, and 40.9% reported decreased motivation to work. Whilst there were changes in the daily work intensity and the level of work-related stress, more than 60% of respondents we sampled did not intend to switch careers. Demographic variables like gender, whether one was a student or an existing healthcare worker, and income related to work motivation. The community's stigma was a negative factor that declined intrinsic motivation as well as decreased work retention. CONCLUSIONS: Our study is instrumental in identifying the impact of COVID-19 on career choices amongst Vietnamese healthcare workers. The factors identified have clear implications for policymaking.
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COVID-19 , Estudantes de Medicina , Humanos , COVID-19/epidemiologia , Atitude do Pessoal de Saúde , Escolha da Profissão , Estudos Transversais , Pandemias , Pessoal de SaúdeRESUMO
This paper presents a new control strategy that combines classical control and an optimization scheme to regulate the output voltage of the bidirectional converter under the presence of matched and mismatched disturbances. In detail, a control-oriented modeling method is presented first to capture the system dynamics in a common canonical form, allowing different disturbances to be considered. To estimate and compensate for unknown disturbances, an extended state observer (ESO)-based continuous sliding mode control is then proposed, which can guarantee high tracking precision, fast disturbance rejection, and chattering reduction. Next, an extremum seeking (ES)-based adaptive scheme is introduced to ensure system robustness as well as optimal control effort under different working scenarios. Finally, comparative simulations with classical proportional-integral-derivative (PID) control and constant switching gains are conducted to verify the effectiveness of the proposed adaptive control methodology through three case studies of load resistance variations, buck/boost mode switching, and input voltage variation.
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Thermogenic adipocytes have potential utility for the development of approaches to treat type 2 diabetes and obesity-associated diseases. Although several reports have proved the positive effect of beige and brown adipocyte transplantation in obese mice, translation to human cell therapy needs improvement. Here, we describe the application of CRISPR activation (CRISPRa) technology for generating safe and efficient adipose-tissue-engineered constructs with enhanced mitochondrial uncoupling protein 1 (UCP1) expression. We designed the CRISPRa system for the activation of UCP1 gene expression. CRISPRa-UCP1 was delivered into mature adipocytes by a baculovirus vector. Modified adipocytes were transplanted in C57BL/6 mice, followed by analysis of grafts, inflammation and systemic glucose metabolism. Staining of grafts on day 8 after transplantation shows them to contain UCP1-positive adipocytes. Following transplantation, adipocytes remain in grafts and exhibit expression of PGC1α transcription factor and hormone sensitive lipase (HSL). Transplantation of CRISPRa-UCP1-modified adipocytes does not influence glucose metabolism or inflammation in recipient mice. We show the utility and safety of baculovirus vectors for CRISPRa-based thermogenic gene activation. Our findings suggest a means of improving existing cell therapy approaches using baculovirus vectors and CRISPRa for modification and transplantation of non-immunogenic adipocytes.
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Tecido Adiposo Marrom , Proteína Desacopladora 1 , Animais , Humanos , Camundongos , Adipócitos Marrons/metabolismo , Tecido Adiposo Marrom/transplante , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Diabetes Mellitus Tipo 2/terapia , Glucose/metabolismo , Camundongos Endogâmicos C57BL , Termogênese/genética , Proteína Desacopladora 1/metabolismoRESUMO
External application of ethanol enhances tolerance to high salinity, drought, and heat stress in various plant species. However, the effects of ethanol application on increased drought tolerance in woody plants, such as the tropical crop "cassava," remain unknown. In the present study, we analyzed the morphological, physiological, and molecular responses of cassava plants subjected to ethanol pretreatment and subsequent drought stress treatment. Ethanol pretreatment induced a slight accumulation of abscisic acid (ABA) and stomatal closure, resulting in a reduced transpiration rate, higher water content in the leaves during drought stress treatment and the starch accumulation in leaves. Transcriptomic analysis revealed that ethanol pretreatment upregulated the expression of ABA signaling-related genes, such as PP2Cs and AITRs, and stress response and protein-folding-related genes, such as heat shock proteins (HSPs). In addition, the upregulation of drought-inducible genes during drought treatment was delayed in ethanol-pretreated plants compared with that in water-pretreated control plants. These results suggest that ethanol pretreatment induces stomatal closure through activation of the ABA signaling pathway, protein folding-related response by activating the HSP/chaperone network and the changes in sugar and starch metabolism, resulting in increased drought avoidance in plants.
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Manihot , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Secas , Etanol/farmacologia , Regulação da Expressão Gênica de Plantas , Proteínas de Choque Térmico/genética , Manihot/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Amido/metabolismo , Estresse Fisiológico/genética , Açúcares/metabolismo , Água/metabolismoRESUMO
KEY MESSAGE: The field survey in this article showed in 'KU50', a popular variety and late-branching type of cassava in Southeast Asia, that flowering rarely occurs in normal-field conditions in Southeast Asia but is strongly induced in the dry season in the mountainous region. Flowering time is correlated with the expression patterns of MeFT1 and homologs of Arabidopsis GI, PHYA, and NF-Ys. Cassava (Manihot esculenta Crantz) is a tropical crop that is propagated vegetatively rather than sexually by seed. Flowering rarely occurs in the erect-type variety grown in Southeast Asia, but it is known that cassava produces flowers every year in mountainous regions. Data pertaining to the effect of environmental factors on flowering time and gene expression in cassava, however, is limited. The aim of the present study was to determine the kinds of environmental conditions that regulate flowering time in cassava and the underlying molecular mechanisms. The flowering status of KU50, a popular variety in Southeast Asia and late-branching type of cassava, was monitored in six fields in Vietnam and Cambodia. At non-flowering and flowering field locations in North Vietnam, the two FLOWERING LOCUS T (FT)-like genes, MeFT1 and MeFT2, were characterized by qPCR, and the pattern of expression of flowering-related genes and genes responsive to environmental signals were analyzed by using RNA sequencing data from time-series samples. Results indicate that cassava flowering was induced in the dry season in the mountain region, and that flowering time was correlated with the expression of MeFT1, and homologs of Arabidopsis GI, PHYA, and NF-Ys. Based upon these data, we hypothesize that floral induction in cassava is triggered by some conditions present in the mountain regions during the dry season.
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Arabidopsis , Manihot , Arabidopsis/genética , Arabidopsis/metabolismo , Sudeste Asiático , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Manihot/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Water scarcity is a serious agricultural problem causing significant losses to crop yield and product quality. The development of technologies to mitigate the damage caused by drought stress is essential for ensuring a sustainable food supply for the increasing global population. We herein report that the exogenous application of ethanol, an inexpensive and environmentally friendly chemical, significantly enhances drought tolerance in Arabidopsis thaliana, rice and wheat. The transcriptomic analyses of ethanol-treated plants revealed the upregulation of genes related to sucrose and starch metabolism, phenylpropanoids and glucosinolate biosynthesis, while metabolomic analysis showed an increased accumulation of sugars, glucosinolates and drought-tolerance-related amino acids. The phenotyping analysis indicated that drought-induced water loss was delayed in the ethanol-treated plants. Furthermore, ethanol treatment induced stomatal closure, resulting in decreased transpiration rate and increased leaf water contents under drought stress conditions. The ethanol treatment did not enhance drought tolerance in the mutant of ABI1, a negative regulator of abscisic acid (ABA) signaling in Arabidopsis, indicating that ABA signaling contributes to ethanol-mediated drought tolerance. The nuclear magnetic resonance analysis using 13C-labeled ethanol indicated that gluconeogenesis is involved in the accumulation of sugars. The ethanol treatment did not enhance the drought tolerance in the aldehyde dehydrogenase (aldh) triple mutant (aldh2b4/aldh2b7/aldh2c4). These results show that ABA signaling and acetic acid biosynthesis are involved in ethanol-mediated drought tolerance and that chemical priming through ethanol application regulates sugar accumulation and gluconeogenesis, leading to enhanced drought tolerance and sustained plant growth. These findings highlight a new survival strategy for increasing crop production under water-limited conditions.
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Arabidopsis , Secas , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Etanol/metabolismo , Regulação da Expressão Gênica de Plantas , Estômatos de Plantas/fisiologia , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico/genética , Açúcares/metabolismo , Água/metabolismoRESUMO
Acidities of lipophilic compounds, such as various ligands or catalysts, in systems consisting of an aqueous phase at equilibrium with a water-immiscible phase (lipid bilayers, phase transfer catalysis, sensor membranes, to name just few) are typically approximated by the aqueous pKa values. Our research shows that such approximations can lead to seriously biased estimations of the acidities as the bulk of solvated H+ ions reside in the aqueous phase, while the lipophilic speciesâboth neutral acid and anionâpredominantly reside in the organic phase. Therefore, the use of aqueous pKa in such situations is not justified. In this work, we provide a more accurate description of the acidities of acids in such systems by applying the biphasic pKa concept. Biphasic pKa values (pKaow values) of 35 acids of various structures and chemical properties were determined in a 1-octanol:water system. We provide detailed descriptions of the UV-vis and NMR measurement methods. The directly obtained (apparent) pKaow values depend on concentration. Concentration-independent values were obtained by extrapolating the apparent values to zero concentration using a Debye-Hückel model.
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Ácidos , Água , Catálise , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância MagnéticaRESUMO
RARA and RXRA contribute to myeloid maturation in both mice and humans, and deletion of Rxra and Rxrb augments leukemic growth in mice. While defining the domains of RXRA that are required for anti-leukemic effects in murine KMT2A-MLLT3 leukemia cells, we unexpectedly identified RXRA DT448/9PP as a constitutively active variant capable of inducing maturation and loss of their proliferative phenotype. RXRA DT448/9PP was associated with ligand-independent activity in reporter assays, with enhanced co-activator interactions, reduced engraftment in vivo, and activation of myeloid maturation transcriptional signatures that overlapped with those of cells treated with the potent RXRA agonist bexarotene, suggestive of constitutive activity that leads to leukemic maturation. Phenotypes of RXRA DT448/9PP appear to differ from those of two other RXRA mutations with forms of constitutive activity (F318A and S427F), in that DT448/9PP activity was resistant to mutations at critical ligand-interacting amino acids (R316A/L326A) and was resistant to pharmacological antagonists, suggesting it may be ligand-independent. These data provide further evidence that activated retinoid X receptors can regulate myeloid maturation and provide a novel constitutively active variant that may be germane for broader studies of retinoid X receptors in other settings.