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1.
Food Chem ; 334: 127519, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32721832

RESUMO

We aimed to characterize and quantify glucosinolate compounds and contents in broccoli, and a total of 80 genotypes and eight developmental organs were analyzed with UHPLC-Triple-TOF-MS. The method was validated in terms of performance, and the coefficients of determination (R2) were 0.97 and 0.99 for glucoraphanin and gluconapin, respectively. In 80 genotypes, twelve glucosinolates were found in broccoli florets ranging from 0.467 to 57.156 µmol/g DW, with the highest glucosinolate content being approximately 122-fold higher than the lowest value. The principal component of glucobrassicin, neoglucobrassicin and glucoraphanin explained 60.53% of the total variance. There were positive correlations among hydroxyglucobrassicin, methoxyglucobrassicin, glucobrassicin, glucoerucin, gluconasturtiin, glucoraphanin, and glucotropaeolin (P < 0.05). The root contained 43% of total glucosinolates in 80 genotypes, and glucoraphanin represented 29% of the total glucosinolate content in different organs. The mutant broccoli genotypes were found by analysis of gluconapin contents in different organs.

2.
Food Chem ; 335: 127638, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-32736158

RESUMO

Using natural antioxidants instead of synthetics ones has been the tendency for retarding the oil deterioration during repeated deep frying process. Concerning this, the comparison between synthetic tertiarybutyl hydroquinone (TBHQ) and rosemary-based antioxidants in frying French fries was hereby evaluated. The quality and stability of frying oils with rosemary-based antioxidants showed higher efficiency than TBHQ regarding oxidation parameters (i.e., chemical indices, sensory, etc.), where rosmarinic acid (RA) was the most effective, followed by rosemary extracts (RE) and carnosic acid (CA). LF-NMR results were highly correlated (R2 = 0.909-0.998) to the change in physicochemical properties tested, where RA could effectively regulate the relaxation spectrum (T2) change and decrease single component relaxation time (T2W). The PCA graph of NIRS also revealed the dynamic change of antioxidant effectiveness in accordance with that obtained by chemical methods. Hence, both LF-NMR and NIRS can be expected as rapid and efficient methods for future monitoring the frying process.

3.
Methods Mol Biol ; 2196: 199-209, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32889722

RESUMO

In eukaryotic cells, the genomic DNA is packaged into chromatin, the basic unit of which is the nucleosome. Studying the mechanism of chromatin formation under physiological conditions is inherently difficult due to the limitations of research approaches. Here we describe how to prepare a biochemical system called yeast nucleoplasmic extracts (YNPE). YNPE is derived from yeast nuclei, and the in vitro system can mimic the physiological conditions of the yeast nucleus in vivo. In YNPE, the dynamic process of chromatin assembly has been observed in real time at the single-molecule level by total internal reflection fluorescence microscopy. YNPE provides a novel tool to investigate many aspects of chromatin assembly under physiological conditions and is competent for single-molecule approaches.

4.
Curr Opin Biotechnol ; 69: 10-16, 2020 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-33032240

RESUMO

Plant-derived diterpenoids are indispensable to plant development, stress-resistance and interaction with environmental microorganisms. Besides significant roles in plant fitness and adaption, many bioactivities beneficial to human beings are also found in diterpenoids from terrestrial plants. However, these high-value compounds are always present in limited species with low-abundance. Complicated chemosynthesis hardly meets the needs of sufficient supplies. To overcome these obstacles, it is necessary to investigate how diterpenoids are biosynthesized in planta, and followed by engineering the biosynthetic pathway to achieve high yield production. This review will summarize the recent progress of plant diterpenoid biosynthetic pathway discovery and engineering, hoping to offer an inspiration for concerned researchers.

5.
Artigo em Inglês | MEDLINE | ID: mdl-33033226

RESUMO

Viruses employ multiple strategies to inhibit host mRNA nuclear export. Distinct to the generally nonselective inhibition mechanisms, ORF10 from gammaherpesviruses inhibits mRNA export in a transcript-selective manner by interacting with Rae1 (RNA export 1) and Nup98 (nucleoporin 98). We now report the structure of ORF10 from MHV-68 (murine gammaherpesvirus 68) bound to the Rae1-Nup98 heterodimer, thereby revealing detailed intermolecular interactions. Structural and functional assays highlight that two highly conserved residues of ORF10, L60 and M413, play critical roles in both complex assembly and mRNA export inhibition. Interestingly, although ORF10 occupies the RNA-binding groove of Rae1-Nup98, the ORF10-Rae1-Nup98 ternary complex still maintains a comparable RNA-binding ability due to the ORF10-RNA direct interaction. Moreover, mutations on the RNA-binding surface of ORF10 disrupt its function of mRNA export inhibition. Our work demonstrates the molecular mechanism of ORF10-mediated selective inhibition and provides insights into the functions of Rae1-Nup98 in regulating host mRNA export.

6.
Pflugers Arch ; 2020 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-33033891

RESUMO

Tacrolimus (TAC, also called FK506), a common immunosuppressive drug used to prevent allograft rejection in transplant patients, is well known to alter the functions of blood vessels. In this study, we sought to determine whether chronic treatment of TAC could inhibit the activity of big-conductance Ca2+-activated K+ (BK) channels in vascular smooth muscle cells (SMCs), leading to hypertension. Our data reveal that the activity of BK channels was inhibited in cerebral artery SMCs (CASMCs) from mice after intraperitoneal injection of TAC once a day for 4 weeks. The voltage sensitivity, Ca2+ sensitivity, and open time of single BK channels were all decreased. In support, BK channel ß1-, but not α-subunit protein expression was significantly decreased in cerebral arteries. In TAC-treated mice, application of norepinephrine induced stronger vasoconstriction in both cerebral and mesenteric arteries as well as a larger [Ca2+]i in CASMCs. Chronic treatment of TAC, similar to BK channel ß1-subunit knockout (KO), resulted in hypertension in mice, but did not cause a further increase in blood pressure in BK channel ß1-subunit KO mice. Moreover, BK channel activity in CASMCs was negatively correlated with blood pressure. Our findings provide novel evidence that TAC inhibits BK channels by reducing the channel ß1-subunit expression and functions in vascular SMCs, leading to enhanced vasoconstriction and hypertension.

7.
Artigo em Inglês | MEDLINE | ID: mdl-33021884

RESUMO

D. segeticola is a significant plant pathogen that causes leaf spot on tea (Camellia sinensis) in Guizhou Province, China, leading to significant losses in production (Zhao et al. 2018). Although genome sequences of D. segeticola have been published (Ren et al. 2019), no data of transcriptome and microRNA of the pathogen and host during infection are available. Here, we report the high-quality transcriptome and microRNA of D. segeticola infection using the platform of Illumina Hiseq 4000 or Illumina Hiseq 2500. The number of annotated unigenes is 10894 and 33932 at several databases for mycelia and tea. PC-3p-9306445_2, nta-MIR6149a-p3_2ss13GA18CG, mdm-MIR535a-p3_2ss13GA18CG, cst-MIR11334-p3_2ss13GA18CG, et al can target more gene, with the number of the target genes being 31, 21, 16 and 11. The sequence of transcriptome and microRNA of D. segeticola will provide an important resource for researchers studying pathogenic mechanism and disease resistance.

8.
Nat Commun ; 11(1): 4928, 2020 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-33004791

RESUMO

High-altitude adaptation of Tibetans represents a remarkable case of natural selection during recent human evolution. Previous genome-wide scans found many non-coding variants under selection, suggesting a pressing need to understand the functional role of non-coding regulatory elements (REs). Here, we generate time courses of paired ATAC-seq and RNA-seq data on cultured HUVECs under hypoxic and normoxic conditions. We further develop a variant interpretation methodology (vPECA) to identify active selected REs (ASREs) and associated regulatory network. We discover three causal SNPs of EPAS1, the key adaptive gene for Tibetans. These SNPs decrease the accessibility of ASREs with weakened binding strength of relevant TFs, and cooperatively down-regulate EPAS1 expression. We further construct the downstream network of EPAS1, elucidating its roles in hypoxic response and angiogenesis. Collectively, we provide a systematic approach to interpret phenotype-associated noncoding variants in proper cell types and relevant dynamic conditions, to model their impact on gene regulation.

9.
Artigo em Inglês | MEDLINE | ID: mdl-33006229

RESUMO

The cell membrane is not only a physical barrier, but also a functional organelle regulating the communication between a cell and its environment. An ability to functionalize the cell membrane with synthetic molecules or nanostructures would advance cellular functions beyond what the evolution of nature has provided. The purpose of this minireview is to introduce recent progress in using synthetic DNA and DNA-based nanostructures for cell surface engineering. We first introduce chemical conjugation and physical binding methods for monovalent and polyvalent surface engineering. We further introduce the application of these methods for either promotion or inhibition of cell-environment communication in numerous applications including promotion of cell-cell recognition, regulation of intracellular pathways, protection of therapeutic cells and sensing of intracellular and extracellular microenvironment. Lastly, we summarize current challenges existing in this area and potential solutions to solve these challenges.

10.
Artigo em Inglês | MEDLINE | ID: mdl-33006530

RESUMO

Colletotrichum species cause anthracnose disease on the economically important spice crop chili. 24 Colletotrichum species are known to infect chili and cause anthracnose. C. scovillei belongs to the C. acutatum species complex, and it shows greater aggressiveness than other species, particularly in the case of inoculation onto the non-wounded fruits of chili plants. The current work introduces an initial Illumina-Nanopore hybrid draft genome for the C. scovillei strain TJNH1 together with the related annotations. Knowledge of this genome sequences provides an important reference genome of C. scovillei and will further help understand the pathogenic mechanism of C.scovillei to plant.

11.
Future Oncol ; 2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-33052751

RESUMO

Aim: To explore the prognostic significance of Syt-7 in hepatocellular carcinoma (HCC) and the potential mechanisms. Methods: Immunohistochemistry was used to examine the expression of Syt-7. Overall survival and disease-free survival were compared between Syt-7 positive and negative groups. The effects of Syt-7 knockdown on BEL-7404 cells were further evaluated. Results: Syt-7 expression was significantly higher in HCC tumorous tissues compared with paracancerous tissues. Syt-7 was closely associated with α-fetoprotein tumor size, vascular invasion, tumor node metastasis stage and tumor differentiation. Syt-7 was an independent risk factor for overall survival and disease-free survival. Additionally, Syt-7 knockdown inhibited proliferation and colony formation and induced cell cycle arrest in HCC cells. Conclusion: Syt-7 overexpression forecasts unfavorable prognosis and promotes cell proliferation in HCC.

12.
BMC Plant Biol ; 20(1): 475, 2020 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-33066728

RESUMO

BACKGROUND: Drought stress is an environmental factor that limits plant growth and reproduction. Little research has been conducted to investigate the MLP gene in tobacco. Here, NtMLP423 was isolated and identified, and its role in drought stress was studied. RESULTS: Overexpression of NtMLP423 improved tolerance to drought stress in tobacco, as determined by physiological analyses of water loss efficiency, reactive oxygen species levels, malondialdehyde content, and levels of osmotic regulatory substances. Overexpression of NtMLP423 in transgenic plants led to greater sensitivity to abscisic acid (ABA)-mediated seed germination and ABA-induced stomatal closure. NtMLP423 also regulated drought tolerance by increasing the levels of ABA under conditions of drought stress. Our study showed that the transcription level of ABA synthetic genes also increased. Overexpression of NtMLP423 reduced membrane damage and ROS accumulation and increased the expression of stress-related genes under drought stress. We also found that NtWRKY71 regulated the transcription of NtMLP423 to improve drought tolerance. CONCLUSIONS: Our results indicated that NtMLP423-overexpressing increased drought tolerance in tobacco via the ABA pathway.

13.
J Stroke Cerebrovasc Dis ; 29(11): 105154, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33066884

RESUMO

OBJECTIVE: Transcranial color-duplex sonography (TCCS) is a promising method in evaluating the hemodynamics in patients with moyamoya disease (MMD). This study aimed to explore the feasibility of preoperative TCCS in predicting the outcome of revascularization surgery in MMD patients. METHODS: We retrospectively analysed 64 cases of MMD patients receiving revascularization surgery from January 2012 to January 2014. We utilized TCCS to perform comprehensive hemodynamic examination on the hemodynamics of bilateral intracranial and extracranial cerebrovascular flow and assessed the surgical outcomes and prognosis through the longitudinal comparison of the preoperative and postoperative cerebrovascular hemodynamics. Occurrence of bypass blockage was regarded as surgical failure. RESULTS: We established a prediction model for bypass blockage among MMD patients with an AUC of 0.858 (95% CI: 0.666-1). The parameters, EDV of ECA and PSV of MA obtained by the model are the main preoperative predictors for bypass blockage. CONCLUSIONS: TCCS could preoperatively determine the degree of MMD and evaluate the outcome of revascularization surgery. It also is a feasible tool to predict the curative effect by providing preoperative hemodynamic information.

14.
Rev Sci Instrum ; 91(9): 095004, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-33003781

RESUMO

A novel piezoelectric rotary motor (PRM) on the basis of synchronized switching control was designed, fabricated, and tested to achieve high speed, high efficiency, and high torque. The new motor mainly consists of a vibrator working in the resonance state as the driving element of the PRM and a clutch working in the quasi-static state to control the shaft for unidirectional rotation. The finite element method software COMSOL Multiphysics 5.4 was used to design the structure of the motor and determine the feasibility of the design mechanism of the PRM. Moreover, an experimental setup was built to validate the working principles and evaluate the performance of the PRM. The prototype motor outputted a no-load speed of 7.21 rpm and a maximum torque of 54.4 N mm at a vibrator driving voltage of 120 Vp-p and a clutch driving voltage of 200 Vp-p. The motor achieved a net efficiency of 15.6% under the preload torque of 3 N mm. The average stepping angle of the motor with no-load was 0.068°, when the voltages applied to the clutch and the vibrator were 200 Vp-p and 120 Vp-p, respectively, with the frequency of 512 Hz.

15.
Biomol Ther (Seoul) ; 2020 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-33041265

RESUMO

Histone acetylation is a well-characterized epigenetic modification controlled by histone acetyltransferases (HATs) and histone deacetylases (HDACs). Imbalanced histone acetylation has been observed in many primary cancers. Therefore, efforts have been made to find drugs or small molecules such as HDAC inhibitors that can revert acetylation levels to normal in cancer cells. We observed dose-dependent reduction in the endogenous and exogenous protein expression levels of KAT8 (also known as human MOF), a member of the MYST family of HATs, and its corresponding histone acetylation at H4K5, H4K8, and H4K16 in chemotherapy drug gemcitabine (GEM)-exposed T24 bladder cancer (BLCA) cells. Interestingly, the reduction in MOF and histone H4 acetylation was inversely proportional to GEM-induced γH2AX, an indicator of chemotherapy drug effectiveness. Furthermore, pGL4-MOF-Luc reporter activities were significantly inhibited by GEM, thereby suggesting that GEM utilizes an MOF-mediated anti-BLCA mechanism of action. In the CCK-8, wound healing assays and Transwell® experiments, the additive effects on cell proliferation and migration were observed in the presence of exogenous MOF and GEM. In addition, the promoted cell sensitivity to GEM by exogenous MOF in BLCA cells was confirmed using an Annexin V-FITC/PI assay. Taken together, our results provide the theoretical basis for elucidating the anti-BLCA mechanism of GEM.

16.
ChemSusChem ; 2020 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-33043594

RESUMO

Electrolytic water splitting using surplus electricity represents one of the most cost-effective and promising strategies for hydrogen production. The high overpotential of the oxygen-evolution reaction (OER) caused by the multi-electron transfer process with a high chemical energy barrier, however, limits its competitiveness. Here, an highly active and stable OER electrocatalyst was designed through a cobalt-induced intrastructural enhancement strategy combined with suitable electronic structure modulation. A carved carbon nanobox was embedded with tri-metal phosphide from a uniform Ni-Co-Fe Prussian blue analogue (PBA) nanocube by sequential NH3•H2O etching and thermal phosphorization. The sample exhibited an OER activity in an alkaline medium, reaching a current density of 10 mA/cm2 at an overpotential of 182 mV and displayed a small Tafel slope of 47 mV/dec, superior to the most recently reported OER electrocatalysts. Moreover, it showed impressive electrocatalytic durability, increasing by approximately 2.7% of operating voltage after 24 h of continuous testing. The excellent OER activity and stability are ascribed to a favorable transfer of mass and charge provided by the porous carbon shell, synergistic catalysis between the three-component metal phosphides originating from appropriate electronic structure modulation, more exposed catalytic sites on the hollow structure, and chainmail catalysis resulting from the carbon protective layer. We foresee that our successfully demonstrated design concept can be easily extended to other heterogeneous catalyst designs.

17.
Sci Rep ; 10(1): 16754, 2020 10 07.
Artigo em Inglês | MEDLINE | ID: mdl-33028909

RESUMO

L-arginine/NOS/NO signaling pathway plays a critical role in controlling variety of vascular diseases. However, whether NOS inhibition by L-NAME suppresses late embryonic development is undefined. The aim of this study is to determine whether NOS inhibition by L-NAME is critical for late embryonic rat hind limb development. The pregnant rat at E13.5 administrated L-NAME by consecutive intraperitoneal injection. The embryos been harvested from E16.5 to E 20.5. Hematoxylin and Eosin Staining, Immunofluorescence and Immunohistochemistry performed to determine hind limb Vasculogenesis, HUVEC culture, Adenoviral PFKFB3 infection, Real time PCR and western blot were performed to determine whether L-arginine/NOS/NO pathway controlling late embryonic hind limb development through PFKFB3 mediated angiogenetic pathway. NOS inhibition by L-NAME resulting in late embryonic hind limb developmental defects characterized by severe hemorrhage. The in vivo studies showed that NOS inhibition strongly suppressed hind limb angiogenetic remodeling by impairing differentiation of endothelial cells and smooth muscle cells, and extracellular matrix synthesis. For underlie mechanism, our studies indicated that L-NAME treatment dramatically suppresses PFKFB3 expression in hematopoietic progenitor cells, tubulogenetic endothelial cells and smooth muscle cells. Knockdown of PFKFB3 dramatically inhibits the expression of angiogenetic genes, as well as tubulogenesis and extracellular matrix related genes. Taken together, our data in this study demonstrated that L-arginine-eNOS-NO pathway is important for rat hind limb development during late embryonic stage. This could be both a useful animal model and a promising therapeutic treatment for defects of late embryonic developmental hind limbs.

19.
Int J Cancer ; 2020 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-33038264

RESUMO

Prostate cancer (PCa) progression is driven by androgen receptor (AR) signaling. Unfortunately, androgen-deprivation therapy and the use of even more potent AR pathway inhibitors (ARPIs) cannot bring about a cure. ARPI resistance (i.e. castration-resistant PCa, CRPC) will inevitably develop. Previously, we demonstrated that GRB10 is an AR transcriptionally-repressed gene that functionally contributes to CRPC development and ARPI resistance. GRB10 expression is elevated prior to CRPC development in our patient-derived xenograft models and is significantly upregulated in clinical CRPC samples. Here, we analyzed transcriptomic data from GRB10 knockdown in PCa cells and found that AR signaling is downregulated. While the mRNA expression of AR target genes decreased upon GRB10 knockdown, AR expression was not affected at the mRNA or protein level. We further found that phosphorylation of AR serine 81 (S81), which is critical for AR transcriptional activity, is decreased by GRB10 knockdown and increased by its overexpression. Luciferase assay using GRB10-knockdown cells also indicate reduced AR activity. Immunoprecipitation coupled with mass spectrometry revealed an interaction between GRB10 and the PP2A complex, which is a known phosphatase of AR. Further validations and analyses showed that GRB10 binds to the PP2Ac catalytic subunit with its PH domain. Mechanistically, GRB10 knockdown increased PP2Ac protein stability, which in turn decreased AR S81 phosphorylation and reduced AR activity. Our findings indicate a reciprocal feedback between GRB10 and AR signaling, implying the importance of GRB10 in PCa progression. This article is protected by copyright. All rights reserved.

20.
Schizophr Res ; 2020 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-33046333

RESUMO

In this study, we applied brain grey matter volume and structural covariance methods on T1 weighted images to delineate potential structural brain changes in individuals with high schizotypy, who were defined as healthy individuals scoring in the top tenth percentile of the Schizotypal Personality Questionnaire (SPQ). Eighty-seven college students with high schizotypy and 122 controls were recruited in China. Differences in grey matter volume and volume covariance between the two groups, and correlations of grey matter volume with SPQ scores in the high schizotypy group were examined. We found that individuals with high schizotypy had decreased grey matter volume at the left medial superior frontal gyrus (medsFG) extending towards the superior frontal gyrus, decreased structural covariance within the right medsFG, between the right superior frontal gyrus (sFG), the right superior temporal gyrus and the right anterior insula; and increased structural covariance between the caudate and the right inferior temporal gyrus. Correlation analysis revealed that grey matter volume of the left middle temporal pole and the right sFG correlated positively with the SPQ total scores, volume of the bilateral cerebellum 9 sub-region correlated negatively with the SPQ cognitive-perceptual sub-scale scores, volume of the bilateral striatum correlated positively with the SPQ interpersonal sub-scale scores, and volume of the bilateral superior temporal pole correlated positively with the SPQ disorganization sub-scale scores in the high schizotypy group. These results highlight important grey matter structural changes in the medsFG in individuals with high schizotypy.

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