Overexpression of PbrGA2ox1 enhances pear drought tolerance through the regulation of GA_(3)-inhibited reactive oxygen species detoxification and abscisic acid signaling

Drought stress is a devastating natural disaster driven by the continuing intensification of global warming,which seriously threatens the productivity and quality of several horticultural crops,including pear.Gibberellins(GAs)play crucial roles in plant growth,development,and responses to drought stress.Previous studies have shown significant reductions of GA levels in plants under drought stress;however,our understanding of the intrinsic regulation mechanisms of GA-mediated drought stress in pear remains very limited.Here,we show that drought stress can impair the accumulation of bioactive GAs(BGAs),and subsequently identified PbrGA2ox1 as a chloroplast-localized GA deactivation gene.This gene was significantly induced by drought stress and abscisic acid(ABA)treatment,but was suppressed by GA_(3)treatment.PbrGA2ox1-overexpressing transgenic tobacco plants(Nicotiana benthamiana)exhibited enhanced tolerance to dehydration and drought stresses,whereas knock-down of PbrGA2ox1 in pear(Pyrus betulaefolia)by virus-induced gene silencing led to elevated drought sensitivity.Transgenic plants were hypersensitive to ABA,and had a lower BGAs content,enhanced reactive oxygen species(ROS)scavenging ability,and augmented ABA accumulation and signaling under drought stress compared to wild-type plants.However,the opposite effects were observed with PbrGA2ox1 silencing in pear.Moreover,exogenous GA_(3)treatment aggravated the ROS toxic effect and restrained ABA synthesis and signaling,resulting in the compromised drought tolerance of pear.In summary,our results shed light on the mechanism by which BGAs are eliminated in pear leaves under drought stress,providing further insights into the mechanism regulating the effects of GA on the drought tolerance of plants.

OsbZIP53 Negatively Regulates Immunity Response by Involving in Reactive Oxygen Species and Salicylic Acid Metabolism in Rice

The basic region/leucine zipper(bZIP)transcription factors play important roles in plant development and responses to abiotic and biotic stresses.OsbZIP53 regulates resistance to Magnaporthe oryzae in rice by analyzing APIP5-RNAi transgenic plants.To further investigate the biological functions of OsbZIP53,we generated osbzip53 mutants using CRISPR/Cas9 editing and also constructed OsbZIP53 over-expression transgenic plants.Comprehensive analysis of phenotypical,physiological,and transcriptional data showed that knocking-out OsbZIP53 not only improved disease resistance by inducing a hypersensitivity response in plants,but also regulated the immune response through the salicylic acid pathway.Specifically,disrupting OsbZIP53 increased H2O2 accumulation by promoting reactive oxygen species generation through up-regulation of several respiratory burst oxidase homologs(Osrboh genes)and weakened H2O2 degradation by directly targeting OsMYBS1.In addition,the growth of osbzip53 mutants was seriously impaired,while OsbZIP53 over-expression lines displayed a similar phenotype to the wild type,suggesting that OsbZIP53 has a balancing effect on rice immune response and growth.

Reactivity Ratios of Diethyldiallylammonium Chloride with Acrylamide or Acrylic Acid

The compositions of copolymers of diethyldiallylammonium chloride （DEDAAC） with acrylamide （AM）, acrylic acid （AA） or sodium acrylic acid （NaAA） at low conversion were determined by elemental analysis, and the reactivity ratios of monomers in copolymerization were obtained by Kelen-Tudos method. The results showed that the reactivity ratios rDE and rAM are 0.31 and 5.27 for DEDAAC with AM, rDE and rAA are 0.28 and 5.15 for DEDAAC with AA, and roe and rNsAA are 0.40 and 3.97 for DEDAAC with NaAA, respectively. The copolymerizations for DEDAAC with AM, AA or NaAA are non-ideal copolymerization and the products are random copolymers.

Behaviour of Tributylamine as Entrainer for the Separation of Water and Acetic Acid with Reactive Extractive Distillation

A new separation method, reactive extractive distillation, was put forward for separating water and acetic acid. The separation mechanism was analyzed through infrared spectra technique. Isobaric vapor-liquid equilibrium (VLE) data at 101.33 kPa for the binary or ternary systems consisting of water, acetic acid and tributylamine were measured. The activity coefficients were correlated by using Wilson, NRTL, and UNIQUAC Equations.The VLE experiment showed that tributylamine could enhance the relative volatility of water to acetic acid. An extractive distillation experiment was carried out and proved that tributylamine was a good extractive solvent.

Variation of reactivity of particulate and sedimentary organic matter along the Zhujiang River Estuary

To investigate organic matter source and reactivity in the Zhujiang River (Pearl River) Estuary and its adjacent areas, particulate organic carbon (POC), particulate hydrolysable amino acids (PHAA), and Chl a during two cruises in July 1999 and July 2000 were measured. The highest POC and PHAA concentration was observed in the waters with maximum Chl a. The spectra distribution, relative content (dry weight in milligram per gram), PHAA-C% POC and other indicators such as the ratios of amino acids vs. amino sugars (AA/AS) and glucosamine vs. galactosamine (Glum/Gal) suggested that particulate amino acids in the water column and sediments in the Zhujiang River Estuary were mainly derived from biogenic processes rather than transported from terrestrial erosion. In inner estuary where high turbidity was often observable, organic matter was mainly contributed by re-suspension of bottom sediments with revealed zooplankton, microbial reworked characteristics, which suggest that these organic matters were relatively 'old'. In the estuarine brackish region, organic matter in water column is mainly contributed by relatively fresh, easily degradable phytoplankton derived organic matter. During physical - biological processes within the eastuary, organic matter derived from phytoplankton was subjected to alteration by zooplankton grazing and bacterial reworking.

Reactive Crystallization of Calcium Sulfate Dihydrate from Acidic Wastewater and Lime

The present work focused on the recycle of the sulfate and the metal ions from acidic wastewater dis charged by nonferrous metallurgical industry. The effects of the temperature, the reactant concentration, the stirring speed and the metal ions on the reactive crystallization process of calcium sulfate between sulfuric acid and lime were systematically investigated. The morphology of the precipitated crystals evolved from plateletlike and nee dlelike shape to rodlike shape when the temperature was increased from 25 to 70 ℃. An increase in the agglom.

Reactive Extraction of Nicotinic Acid with Trialkylamine in n-Octanol

The distributions of nicotinic acid (NA) between water and trialkylamine (N235) dissolved in n-octanol was studied. The complexes of N235 and NA were investigated by Fourier transformation infrared spectrometry to deduce the reaction mechanism. It was found that N235/n-octanol was an efficient extractant for extracting nicotinic acid. The favorable operation conditions were equilibrium aqueous pH 4.2 to 5.5 and initial N235 concentration<0.42 mol·L-1 . The reaction processes included the reaction between neutral N235 and neutral NA and the reaction between protonated N235 and anionic NA. Based on the mass action law and some assumptions, an expression for distribution coefficient D was proposed. The apparent extraction equilibrium constants were calculated by fitting the experimental data and the results were satisfactory.

All-trans retinoic acid increases ARPE-19 cell apoptosis via activation of reactive oxygen species and endoplasmic reticulum stress pathways

AIM:To explore the apoptosis of ARPE-19 cells after the treatment with different doses of all-trans-retinoic acid(ATRA).METHODS:ARPE-19 cells were used in the in-vitro experiment.Flow cytometry assay was employed to evaluate the level of reactive oxygen species(ROS)and apoptosis.The effects of ATRA(concentrations from 2.5 to 20μmol/L)on the expression of endoplasmic reticulum stress(ERS)markers in vitro were evaluated by Western blot and realtime quantitative polymerase chain reaction(qRT-PCR)assays.The contribution of ROS and ERS-induced apoptosis in vitro was determined by using N-acetyl-L-cysteine(NAC)and Salubrinal,an antagonist of NAC and ERS,respectively.RESULTS:Flow cytometry showed that ATRA significantly increased ARPE-19 cell apoptosis and ROS levels in each group(F=86.39,P<0.001;F=116.839.P<0.001).Western blot and qRT-PCR revealed that levels of CHOP and BIP were elevated in a concentration-dependent pattern after the cells were incubated with ATRA(2.5-20μmol/L).The upregulation of VEGF-A and CHOP induced by ATRA could be inhibited by NAC(antioxidant)and Salubrinal(ERS inhibitor)in vitro.CONCLUSION:ATRA induces the apoptosis of ARPE-19 cells via activated ROS and ERS signaling pathways.

Application of Bronsted acid ionic liquids as green catalyst in the synthesis of 2-propanol with reactive distillation

Five Br?nsted acidic ionic liquids(ILs) were prepared and characterized by FT-IR,~1H NMR and ^(13)C NMR. Their catalytic activities for the synthesis of 2-propanol(IPOH) via transesterification of isopropyl acetate(IPAc) with methanol(Me OH) were investigated. Among all the tested ILs, [Ps-mim]HSO_4 performed best and was used as catalyst for further studies. The reaction kinetics were carried out to correlate the parameters in a homogeneous second order kinetic model. It has been found that there is close agreement between the calculated and experimental values. The high-pressure batch reactive distillation experimental apparatus was set up in order to enhance the conversion of IPAc. A high conversion of IPAc of 99.4% was obtained under the optimal reaction conditions. The catalyst [Ps-mim]HSO_4 can be recycled easily by a rotary evaporator and reused without any further treatment. The catalyst had been repeatedly used for four times and no obvious changes in the structure of catalyst could be observed.

Performance of Metal and Acid Ions Remediation in Contaminated Soils by Electrokinetics with Chitosan Permeable Reactive Barrier

Metal and acid ions contamination of soil in China is serious. To find an efficient solution for remediating the combined pollution,electrokinetics( EK) coupled with chitosan( CTS)permeable reactive barrier( EK/CPRB) was used to investigate the performances of metal and acid ions remediation. Adsorption characteristics of Zn^(2+),Fe^(3+),Ca^(2+),SO_4^(2-) and NO_3^- onto CTS were also conducted. The results showed the sorption of Zn^(2+),Fe^(3+),Ca^(2+),SO_4^(2-) and NO_3^- on CTS could be well described by Freundlich model. When the CTS dosage is 8 g,the total removal efficiency for Zn^(2+),Fe^(3+),Ca^(2+),SO_4^(2-) and NO_3^- is 86. 8%,90. 2%,92. 4%,90. 0% and 82. 5%,respectively. CTS enhanced ions remediation efficiencies significantly compared with the single EK system,especially for SO_4^(2-) and NO_3^-. The results indicate EK/CPRB system is suitable for the remediation of soil contaminated by both metal ions and acid ions.

Reactive Compatibilization of Short-Fiber Reinforced Poly(lactic acid)Biocomposites

Poor interfacial adhesion between biobased thermoplastics and natural fibers is recognized as a major drawback for biocomposites.To be applicable for the large-scale production,a simple method to handle is of importance.This work presented poly(lactic acid)(PLA)reinforced with short-fiber and three reactive agents including anhydride and epoxide groups were selected as compatibilizers.Biocomposites were prepared by one-step meltmixing methods.The influence of reactive agents on mechanical,dynamic mechanical properties and morphology of PLA biocomposites were investigated.Tensile strength and storage modulus of PLA biocomposites incorporated with epoxide-based reactive agent was increased 13.9%and 37.4%compared to non-compatibilized PLA biocomposite,which was higher than adding anhydride-based reactive agent.SEM micrographs and Molau test exhibited an improvement of interfacial fiber-matrix adhesion in the PLA biocomposites incorporated with epoxide-based reactive agent.FTIR revealed the chemical reaction between the fiber and PLA with the presence of epoxide-based reactive agents.

Intensified reactive extraction of 4-hydroxypyridine with di(2-ethylhexyl) phosphoric acid in 1-octanol by using tributyl phosphate

The efficient separation of amphoteric organic compounds from dilute solutions is of great importance in the industrial field. In the present work, the reactive extractions of 4-hydroxypyridine(4-HP) with tributyl phosphate(TBP), di(2-ethylhexyl) phosphoric acid(D2EHPA) and TBP + D2EHPA dissolved in 1-octanol were investigated, respectively. The influences of the initial concentrations of TBP, D2EHPA and TBP + D2EHPA on distribution ratio(D) were discussed, as well as the reactive extraction mechanism were proposed. The obvious intensification effect was observed when the mixture of TBP and D2EHPA was used as extractant. The best extraction conditions were found to be of the molar ratio of D2EHPA and TBP at 2:1 and the equilibrium aqueous pH at 3.50-4.50. D values increased with the increase of the total concentration of TBP and D2EHPA in 1-octanol. Especially, the analysis on the extraction mechanisms clearly indicate(i) TBP in 1-octanol shows negligible reactive extraction toward 4-HP,(ii) D2EHPA in 1-octanol exhibits moderate extraction effect by forming 4-HP:D2EHPA(1:1) and 4-HP:2D2EHPA(1:2) type complexes, while(iii) D2EHPA in TBP/1-octanol demonstrates the maximum distribution ratio with the 4-HP:D2EHPA(1:1) type complex domination. The discussion provides new insights on the mechanism and opens a new way for the intensified extraction of amphoteric organic compounds by using the mixture of multiple extractants in the diluent.

Dichloroacetic Acid (DCA)-Induced Cytotoxicity in Human Breast Cancer Cells Accompanies Changes in Mitochondrial Membrane Permeability and Production of Reactive Oxygen Species

Cancer cells utilize cytosolic glycolysis for their energy production even in the presence of adequate levels of oxygen (Warbug effect) due to mitochondrial defects. Dichloroacetic acid (DCA) shifts cytosolic glucose metabolism to aerobic oxidation by inhibiting mitochondrial pyruvate dehydrogenase kinase (PDK) and increasing pyruvate uptake. Therefore, DCA has potential in reversing the glycolytic metabolism defect in cancerous cells. DCA is also known to induce apoptosis in a number of cancer cell lines, the mechanism of which is not well understood. In this study, an attempt has been made to investigate the effects of DCA on aggressive human breast cancer (MCF-7) cells as compared with less aggressive mouse osteoblastic (MC3T3) cells. Cell cytotoxicity was determined by MTT, crystal violet and Trypan blue exclusion assays. Western blot was used to detect any changes in the expression of apoptotic markers. Flow cytometry was used to measure apoptotic and necrotic effects of DCA. Mitochondrial integrity was determined by change in mitochondrial membrane potential (Δψm), whereas oxidative damage was determined by production of reactive oxygen species (ROS). DCA caused a concentration-dependent cytotoxicity both in MCF-7 and MC3T3 cell lines. MCF-7 cells were most affected. Flow cytometry results showed a significantly higher apoptosis in MCF-7 even at lower concentrations of DCA. However, higher concentrations of DCA were necrotic. Western blotting showed an increased expression of Mn-SOD-1 upon DCA treatment. Further, DCA decreased Δψm and increased ROS production. The effects of DCA were more pronounced on MCF-7 cells as compared to MC3T3 cells. Our results suggest that DCA-induced cytotoxicity in cancerous cells is mediated via changes in Δψm and production of ROS.

Equilibrium Study for Reactive Extraction of Caproic Acid in Mibk and Xylene

The recovery of caproic acid from the aqueous stream is of practical relevance due to its market value and the stringent environmental standards. The profound success of reactive extraction has been noted in the recovery of carboxylic acids from dilute solutions. In present paper, equilibrium of caproic acid with aliquat-336, a quaternary amine compound, in xylene and Methyl Isobutyl Ketone (MIBK) has been studied. Equilibrium parameters such as distribution coefficients, loading ratio, degree of extraction and equilibrium complexation constants have been presented. The data obtained is useful in understanding the equilibrium characteristics and efficient design of the recovery process of caproic acid by reactive extraction.

Separation of Lactic Acid from Diluted Solution by Hybrid Short Path Evaporation and Reactive Distillation

This work describes the separation and purification of lactic acid from diluted solution by HSPE （hybrid short path evaporation） and RD （reactive distillation） as coupled process. The results showed that it is possible to increase lactic acid concentration up to 4.7 times higher than the raw material concentration.

Gossypol acetic acid regulates leukemia stem cells by degrading LRPPRC via inhibiting IL-6/JAK1/STAT3 signaling or resulting mitochondrial dysfunction

BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.

Protective effect of long-chain polyunsaturated fatty acids on hepatorenal syndrome in rats

BACKGROUND Hepatorenal syndrome(HRS)is the most prevalent form of acute kidney injury in cirrhotic patients.It is characterized by reduced renal blood flow and represents the most severe complication in cirrhotic patients with advanced disease.Previous research has indicated that antioxidants can delay the onset of a hyperdynamic circulatory state in cirrhosis and improve renal function in HRS patients.Regular omega-3 supplementation has significantly reduced the risk of liver disease.This supplementation could represent an additional therapy for individuals with HRS.AIM To evaluated the antioxidant effect of omega-3 polyunsaturated fatty acid supplementation on the kidneys of cirrhotic rats.METHODS Secondary biliary cirrhosis was induced in rats by biliary duct ligation(BDL)for 28 d.We used 24 male Wistar rats divided into the following groups:I(control);II(treated with omega-3,1 g/kg of body weight);III(BDL treated with omega-3,1 g/kg of body weight);and IV(BDL without treatment).The animals were killed by overdose of anesthetic;the kidneys were dissected,removed,frozen in liquid nitrogen,and stored in a freezer at-80℃for later analysis.We evaluated oxidative stress,nitric oxide(NO)metabolites,DNA damage by the comet assay,cell viability test,and apoptosis in the kidneys.Data were analyzed by one-way analysis of variance,and means were compared using the Tukey test,with P≤0.05.RESULTS Omega-3 significantly decreased the production of reactive oxygen species(P<0.001)and lipoperoxidation in the kidneys of cirrhotic rats treated with omega-3(P<0.001).The activity of the antioxidant enzymes superoxide dismutase and catalase increased in the BDL+omega-3 group compared to the BDL group(P<0.01).NO production,DNA damage,and caspase-9 cleavage decreased significantly in the omega-3-treated BDL group.There was an increase in mitochondrial electrochemical potential(P<0.001)in BDL treated with omega-3 compared to BDL.No changes in the cell survival index in HRS with omega-3 compared to the control group(P>0.05)were observed.CONCLUSION The study demonstrates that omega-3 can protect cellular integrity and function by increasing antioxidant enzymes,inhibiting the formation of free radicals,and reducing apoptosis.

Research on Oxidative Stress Induced by Tenuazonic Acid from Alternaria augustiovoide and Changes in Antioxidant Enzyme Activities in Leaves of Echinochloa crus-galli

[Objective] The aim of the study was to determine whether phytotoxicity of TeA against Echinochloa crus-galli leaves was correlated with oxidative stress caused by generation of reactive oxygen and the changes of antioxidant enzymes activity.[Method] The changes of malondialdehyde（MDA）content,hydrogen peroxide（H2O2）,and activities of superoxide dismutase（SOD）,glutathione reductase（GR）and catalase（CAT）were studied by leaf segment method in vitro.[Result] After the treatment of 500 μmol/L TeA,the content of MDA and H2O2 increased by 247.86% and 67.00%,respectively,indicating that the accumulation of MDA and H2O2 in E.crus-galli leaves was due to the reactive oxygen burst induced by TeA.TeA induced a significant increase in activities of SOD,GR and CAT.At 500 μmol/L TeA,activities of SOD,GR and CAT increased more than one fold compared with the control.[Conclusion] TeA could not only cause oxidative stress in leaves of E.crus-galli through the induction of reactive oxygen,but also induce the increasing of antioxidant enzyme activity.

鼠尾草酸影响线粒体功能抑制破骨细胞分化

背景:鼠尾草酸是在迷迭香中发现的一种生物活性化合物,已被证明可减少炎症和活性氧,但在破骨细胞分化进程中的作用机制尚不明确。目的:探讨鼠尾草酸对破骨细胞活化、活性氧产生及线粒体功能的影响。方法:体外提取培养小鼠来源的原代骨髓源巨噬细胞,使用CCK-8细胞活力实验检测不同浓度(0,10,15,20,25和30μmol/L)鼠尾草酸对骨髓源巨噬细胞的增殖和毒性作用,筛选出安全作用浓度。将骨髓源巨噬细胞按照浓度梯度分组培养,核因子κB受体活化因子配体诱导破骨细胞5-7 d后,分别进行抗酒石酸酸性磷酸酶染色、F-actin染色、H2DCFDA探针和线粒体活性氧、Mito-Tracker荧光检测,观察鼠尾草酸对破骨细胞分化和功能的影响;通过Western Blot及RT-PCR实验检测鼠尾草酸对核因子κB受体活化因子配体诱导的丝裂原激活蛋白激酶信号通路上下游基因和蛋白的影响。结果与结论:①抗酒石酸酸性磷酸酶和F-actin染色显示:鼠尾草酸以浓度依赖性地抑制体外破骨细胞分化及细胞骨架肌动蛋白环形成,其中鼠尾草酸30μmol/L组的抑制作用最显著;与其他干预时期相比,鼠尾草酸在破骨分化的早期(第1-3天)抑制作用最显著;②H2DCFDA探针和线粒体活性氧、Mito-Tracker荧光显示:鼠尾草酸抑制细胞活性氧和线粒体内活性氧的产生,同时减少线粒体膜电位,影响线粒体的功能;③Western Blot及RT-PCR结果显示:鼠尾草酸可抑制与破骨分化相关的NFATc1、CTSK、MMP9、C-fos蛋白的表达,下调与破骨分化相关的NFATc1、Atp6vod2、ACP5、CTSK和C-fos基因的表达;鼠尾草酸还可以增强抗氧化酶蛋白的表达,减少破骨分化过程中活性氧的产生;④鼠尾草酸抑制P38/ERK/JNK蛋白的磷酸化修饰,通过激活MAPK信号通路抑制骨髓源巨噬细胞破骨细胞分化。